ISSN:1 2359-4721

Official Journal of the Brazilian Society of Toxicology

Volume 2 / Suppl. 2 / 2017

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ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017

Applied Research in Toxicolgy the official journal of the Brazilian Society of Toxicology, publishes peer-reviewed original scientific research in all fields of toxicology, including but not limited to nanotoxicology, genomics and proteomics, teratogenesis, carcinogenesis, mutagenesis, reproductive and endocrine toxicology, toxicopathology, target organ toxicity, neurobehavioral toxicology, mechanistic studies, biochemical and molecular toxicology, novel biomarkers, risk assessment and environmental health studies. Manuscripts on clinical toxicology and its aspects, toxicodynamics and toxicokinetics are also accepted. In addition to original research articles, concise and current review and mini-review articles are also welcome, as are case report papers.

Board of Editors

E D I T O R - I N - C H I E F

Daniel J. Dorta Brazilian Society of Toxicology University of São Paulo Ribeirão Preto, Brazil

E D I T O R S

Silvia B. M. Barros Brazilian Society of Toxicology São Paulo, Brazil

Eduardo M. De Capitani Brazilian Association of Information Centers and Toxicological Assistance and Clinical Toxicologists Campinas, Brazil

Gisela A. Umbuzeiro Brazilian Association of Mutagenesis and Environmental Genomics—MutaGen-Brasil Campinas, Brazil

A S S O C I A T E - E D I T O R S

Bruno Megarbane Carlos M. Palmeira Danielle Palma de Oliveira University of Paris University of Coimbra University of São Paulo Paris, France Coimbra, Portugal Ribeirão Preto, Brazil

João Lauro Viana de Camargo Mauricio Yonamine State University Paulista University of São Paulo São Paulo, Brazil São Paulo, Brazil

E D I T O R I A L B O A R D

Bruno Spinosa de Martinis - University of São Paulo - Ribeirão Preto, Brazil

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Brazilian Society of Toxicology

Board of Directors (2016– 2017)

President: Danielle Palma de Oliveira (SP) Vice-President: Maurício Yonamine (SP) General Secretary: Solange Cristina Garcia (RS) 1st Secretary: Tiago Franco de Oliveira (SP) 2nd. Secretary: Paula da Silva Kujbida (RN) 1st. Treasurer: Rafael Menck de Almeida (SP) 2nd. Treasurer: Gisele Augusto Rodrigues de Oliveira (GO)

Advisory Councillors

(2016-2017 Term of office) (2016-2019 Term of office) - Bruno Spinosa de Martinis - Larissa Helena Lobo Torres - Sarah C. Werner de Souza Eller - Emerson Luiz Botelho Lourenço - Elisa Raquel Anastácio Ferraz - Daniel Junqueira Dorta

Fiscal Committee (2016-2017 Term of office)

Titular Substitutes - Éric Diego Barioni - Daniela Morais Leme - Raphael Caio Tamborelli Garcia - Ernani Pinto Júnior - Rony Anderson Rezende Costa - Tânia Marcourakis S 4

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017

XX BRAZILIAN CONGRESS OF TOXICOLOGY (XX CBTox) October 8 - 11, 2017 Federal University of Goiás (UFG)

http://www.cbtox2017.com.br

ABSTRACT BOOK “21st Century Toxicology Strategies for a Safer and More Sustainable World”

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ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017

XX CBTox PROMOTION / SUPPORT / SPONSORS:

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ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017

XX BRAZILIAN CONGRESS OF TOXICOLOGY (XX CBTox)

Organizing Committee Luiz Carlos da Cunha (UFG - GO) Gisele Augusto Rodrigues de Oliveira (UFG – GO) Marize Campos Valadares (UFG – GO) Elisa Raquel Anastácio Ferraz Avelino (UFF – RJ) Frank Sousa Castro (PUC – GO) Flávia Neri Meira de Oliveira (Secretaria da Saúde do Distrito Federal – DF) Sérgio Henrique Nascente Costa (UFG / PUC – GO) Vânia Cristina Rodríguez Salazar (PUC – GO)

Scientific Committee Gisele Augusto Rodrigues de Oliveira (UFG – GO) Daniel Junqueira Dorta (USP – SP) Daniela Morais Leme (UFPR – PR) Danielle Palma de Oliveira (USP – SP) Elisa Raquel Anastácio Ferraz Avelino (UFF – RJ) Lusânia Maria Greggi Antunes (USP – SP) Luiz Carlos da Cunha (UFG – GO) Marize Campos Valadares (UFG – GO) Maurício Yonamine (USP – GO) Rhaul de Oliveira (UNICAMP / USP – SP)

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ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017

PROF. ESTER DE CAMARGO FONSECA MORAES AWARD

This award is given every two years during the Brazilian Congress of Toxicology, and it is the ultimate recognition of the Brazilian Society of Toxicology for people with a long history of distinguished contributions to the Brazilian Toxicology.

We are pleased to announce:

Prof. Dr. Dermeval de Carvalho

As the 2017 honoree with the Award

Dr. Dermeval de Carvalho has a PhD in Pharmaceutical Sciences by the University of São Paulo; Full Professor of Toxicological Analysis at University of São Paulo. Since 1995 he has dedicated himself to Regulatory Toxicology and the safety assessment of ingredients and cosmetic products, besides implanting alternative methods to animal experimentation.

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1R INSTITUTE / XX CBTox AWARDS

- Ana Rosa Lins de Souza - Ana Carolina Conchon Costa - Anne Caroline Cezimbra da Silva - André Bittencourt Martins - Camila Braggion - Andréia da Silva Fernandes Campos - Clara Mariana Gonçalves Lima - Antonio Anax Falcão de Oliveira - Gabriela de Fatima da Silva Soares - Daniela Barbosa Rocha - Jamylle Martins - Farah Maria Drumond Chequer Baldoni - Orlando Ronaldy da Silva Santos - José Roberto Santin - Rayza Rocha Barraza Aspeé - Michelle Sabrina da Silva - Tâmela Zamboni Madaloz - Raul Ghiraldelli Miranda - Wilma Karlla Azarias Florêncio da Silva - Renata Spagolla Napoleão Tavares - Silvia Romano De Assis - Tatiana Do Nascimento Pedrosa - Vanusa Maria Bispo PROGRAMAÇÃO OFICIAL

08 DE OUTUBRO DE 2017 - DOMINGO HORÁRIO ATIVIDADE TEMA PALESTRANTE (INSTITUIÇÃO) SALA IV FÓRUM DOS CENTROS DE INFORMAÇÃO E ASSISTÊNCIA TOXICOLÓGICA – CIATox ESTHER CAMARGO DE FONSECA 8h–17H REUNIÃO 01 Chair: Dra. Edna Maria Miello Hernandez (Centro de Eventos da UFG) Secretaria Municipal Saúde de São Paulo (COVISA / SMS - SP) ABRACIT Análise e discussão da utilização do sistema de registro DATATOX e 8h – 10h R1.1 Representantes dos CIATox das bases de dados TOXBASE e Micromedex pelos CIATox

10h – 10h20 INTERVALO Elaboração das Diretrizes Diagnósticas e Terapêuticas para Coordenação Geral de Vigilância em Saúde R1.2 10h20 - 11h20 intoxicações por agrotóxicos no Brasil Ambiental (CGVAM) Protocolos de Acidentes·por Animais Peçonhentos e a disponibilidade de Departamento de Vigilância das Doenças 11h20 - 12h R1.3 Soro Antiveneno Transmissíveis (DEVIT-SVS/MS)

12h – 14h ALMOÇO A lmplementação dos Centros de lnformação e Assistência Toxicológica (CIATox)' 14h – 14h45 R1.4.1 com estabelecimentos de saúde integrantes da Rede de Atenção às Urgências Ministério da Saúde/SAS e Emergências no âmbito do Sistema Único de Saúde – SUS O papel Sentinela dos Centros de lnformação e Assistência Toxicológica 14h45 - 15h25 R1.4.2 ANVISA (CIATox) do Brasil A construção dos parâmetros mínimos para o funcionamento dos 15h25 – 16h R1.4.3 ABRACIT CIATox e acreditação

16h – 16h30 R1.5 DEBATE Produção de relatório nal com encaminhamentos 16h30 - 17h R1.6 Encerramento

TOXICOLOGIA CLÍNICA DE URGÊNCIA MARIA ELISA DE SIQUEIRA 8h – 12h MINICURSO 01 Chair: Dr. Luiz Carlos da Cunha (Centro de Eventos da UFG) Universidade Federal de Goiás (UFG)

Intoxicações Agudas: Diagnóstico Carlos Augusto Melo da Silva 8h – 8h40 MC 1.1 e Manejo Inicial TOXIKONSULT – RS Ms. Sergio Emmanuele Graff 8h40 – 9h30 MC 1.2 Intoxicações Agudas por Inseticidas TOXICLIN - SP 9h30 – 10h MC 1.3 DISCUSSÃO

10h – 10h20 INTERVALO

Carlos Augusto Melo da Silva 10h20 – 11h00 MC 1.4 Intoxicações Agudas por Medicamentos TOXIKONSULT – RS Ms. Sergio Emmanuele Graff 11h00 – 11h40 MC 1.5 Intoxicações Agudas por Herbicidas TOXICLIN – SP 11h40 – 12h MC 1.6 DISCUSSÃO NANOTOXICOLOGY MINICURSO 02 MARIA ELISA DE SIQUEIRA 13h – 17h Chair: Dra. Juliana Cancino-Bernardi (em inglês) (Centro de Eventos da UFG) Universidade de São Paulo (USP) Overview of principles and approaches for hazard assessment Dr. Bengt Fadeel 13h – 13h50 MC 2.1 of engineered nanomaterials Karolinska Institutet, Suécia Regulatory research in nanotechnology: How to provide the right Dr. Ary Correa Júnior 13h50 – 14h40 MC 2.2 answers to National Regulation and Legislation Authorities? Universidade Federal de Minas Gerais (UFMG)

14h40 – 15h MC 2.3 DISCUSSÃO

Chair: Dr. Ary Correa Júnior Universidade Federal de Minas Gerais (UFMG)

Dr. Joachim Sturve Ecotoxicology of nanoparticles 15h – 15h50 MC 2.4 University of Gothenburg, Suécia Dra. Juliana Cancino-Bernardi 15h50 – 16h40 MC 2.5 Analytical and alternative methods to assess nanoparticle toxicology Universidade de São Paulo (USP)

16h40 – 17h MC 2.6 DISCUSSÃO

PUBLISHING IN TOXICOLOGY: TIPS AND TRICKS MINICURSO 03 SILVIA BERLANGA BARROS 8h – 17h Chair: Dr. Daniel Junqueira Dorta (em inglês) (Centro de Eventos da UFG) Universidade de São Paulo (FFCLRP/USP) Dra. Wilma de Grava Kempinas 8h – 8h40 MC 3.1 Introduction Universidade Estadual Paulista (UNESP) Dr. Warren Foster How to prepare a good manuscript 8h40 – 10h MC 3.2 McMaster University - Health Sciences Centre, Canadá

10h – 10h20 INTERVALO Dr. Daniel Cyr 10h20 – 12h MC 3.3 What are the features and general endpoints of a good manuscript in Toxicology? Institut national de la recherche scientique (INRS, Canadá)

12h – 14h ALMOÇO Chair: Dra. Wilma de Grava Kempinas Universidade Estadual Paulista (UNESP)

Dr. Thomas Knudsen Publication Challenges in Big-Data Mining and Biological Modeling 14h - 15h30 MC 3.4 Environmental Protection Agency (EPA, EUA) Dr. Fernando Barbosa Junior 15h – 16h MC 3.5 Ecotoxicology and Environmental Safety Universidade de São Paulo (FCFRP/USP) Dr. Daniel Junqueira Dorta 16h – 17h MC 3.6 Applied Research in Toxicology Universidade de São Paulo (FFCLRP/USP)

A TOXICOLOGIA E A QUÍMICA FORENSE: DA COLETA AO LAUDO ALICE DA MATTA CHASIN 8h - 17h MINICURSO 04 Chair: Dr. José Luiz da Costa (Centro de Eventos da UFG) Universidade Estadual de Campinas (UNICAMP) / CIATox-Campinas

Msc. Elvis Medeiros de Aquino SPTC-SP Análise de drogas de abuso em Química Forense - as etapas dos exames 8h – 10h MC 4.1 MSc. Luciano Arantes pericial de substâncias apreendidas Perito Criminal da Polícia Civil do Distrito Federal (SPAL/PCDF) 10h – 10h20 INTERVALO

Sistema de Alerta Prévio (Early Warning System) e seu papel no Dr. Rita Jorge MC 4.2 10h20 – 11h20 monitoramento de Novas Substâncias Psicoativas European Monitoring Centre for Drugs and Drug Addiction (EMCDDA, Portugal) Determinação de drogas de abuso por GC-MS – a importância Msc. Flávia Lopes Roveri 11h20 – 12h MC 4.3 do eciente preparo de amostras Shimadzu do Brasil 12h – 14h ALMOÇO

Chair: MSc. Luciano Arantes Perito Criminal da Polícia Civil do Distrito Federal (SPAL/PCDF)

Dr. Rafael Lanaro 14h – 15h MC 4.4 A Toxicologia Forense realizada em indivíduos vivos CIATox-Campinas Dr. José Luiz da Costa 15h -16h MC 4.5 Toxicologia Forense post mortem Universidade Estadual de Campinas (UNICAMP) / CIATox-Campinas Como obter análises de drogas de abuso, hipnóticos, psicotrópicos, prescritos, Dr. Daniel Saidemberg 16h – 17h MC 4.6 pesticidas e toxinas naturais com conrmação espectral em uma única análise Shimadzu do Brasil

APLICAÇÕES DO EMBRIÃO DE ZEBRAFISH EM TOXICOLOGIA MINICURSO 05 SALA 01, BIOTÉRIO e LPF01 8h - 17h Chair: Dra. Gisele A. Rodrigues de Oliveira (teórico-prático) (Faculdade de Farmácia – UFG) Universidade Federal de Goiás (UFG)

Dra. Gisele A. Rodrigues de Oliveira (FF/UFG) 8h – 9h MC 5.1 Breve apresentação: os palestrantes, os participantes e o zebrash. Dr. Rhaul de Oliveira Universidade de São Paulo (FCF/USP) A ética no uso de animais em experimentação cientíca: o biotério 9h – 10h MC 5.2 Dra. Gisele A. Rodrigues de Oliveira (FF/UFG) de zebrash da FF/UFG

10h – 10h15 INTERVALO O teste de toxicidade em embriões de zebrash: Fish Embryo Toxicity 10h15 – 11h MC 5.3 Dra Thayres de Sousa Andrade (IB/UnB) (FET) test (OECD 236) Dr. Rhaul de Oliveira 11h – 12h MC 5.4 Teórico-prática: cultivo e ensaios com embriões de zebrash (parte 1) Universidade de São Paulo (FCF/USP) Dra Thayres de Sousa Andrade (IB/UnB)

12h – 13h ALMOÇO Dr. Rhaul de Oliveira 13h – 14h MC 5.5 Teórico-prática: cultivo e ensaios com embriões de zebrash (parte 2) Universidade de São Paulo (FCF/USP) Dra. Thayres de Sousa Andrade (IB/UnB) Dr. Rhaul de Oliveira 14h – 15h MC 5.6 Estudos de caso: a avaliação da toxicidade de fármacos e praguicidas. Universidade de São Paulo (FCF/USP) Dra. Thayres de Sousa Andrade (IB/UnB)

15h – 15h15 INTERVALO Dr. Rhaul de Oliveira Atividade Prática: Leitura de teste de toxicidade com 15h15 – 16h MC 5.7 Universidade de São Paulo (FCF/USP) embriões e larvas de zebrash Dra Thayres de Sousa Andrade (IB/UnB) Dr. Rhaul de Oliveira 16h – 16h30 MC 5.8 Encerramento e avaliação do minicurso Universidade de São Paulo (FCF/USP) Dra Thayres de Sousa Andrade (IB/UnB)

MINICURSO 06A ALTERNATIVE METHODS TO EVALUATION OF TOXICITY: TRAINING BY IIVS ANFITEATRO E LABORATÓRIO LFTC/FARMATEC 8h - 12h (teórico-prático / em inglês) Chair: Dra. Marize Campos Valadares (Faculdade de Farmácia – UFG) Universidade Federal de Goiás (UFG) Msc. Allison Hilberer Institute For In Vitro Sciences (IIVS, EUA) 8h – 10h30 MC 6A.1 Bovine Cornea Opacity/Permeability test (BCOP - OECD 437) Dr. Rodger Curren Institute For In Vitro Sciences (IIVS, EUA) Msc. Allison Hilberer 3T3 NRU Phototoxicity Test Institute For In Vitro Sciences (IIVS, EUA) 10h30 – 12h MC 6A.2 (OECD 432) Dr. Rodger Curren Institute For In Vitro Sciences (IIVS, EUA)

APLICAÇÃO DO MODELO SKINETHIC™ RHE PARA CLASSIFICAÇÃO DE COMPOSTOS IRRITANTES E CORROSIVOS MINICURSO 06B ANFITEATRO E LABORATÓRIO LFTC/FARMATEC 13h - 17h Chair: Dra. Marize Campos Valadares (teórico-prático / em inglês) (Faculdade de Farmácia – UFG) Universidade Federal de Goiás (UFG)

Msc. Rodrigo De Vecchi e L'oreal Brasil (Episkin Academy) Aplicação do modelo Skinethic™ RHE para classicação de compostos irritantes e corrosivos 13h – 17h MC 6B.1 Instrutores: Guilherme Mattos e Vanja Dakic (Episkin Academy)

CERIMÔNIA DE ABERTURA ESTHER CAMARGO DE FONSECA 18h30 – 19h30 BANDA PEQUI (Projeto de Extensão e Cultura da Escola de Música da UFG) Prêmio Profa. Ester de Camargo Fonseca Moraes (SBTox) (Centro de Eventos da UFG) PALESTRA DE ABERTURA (em inglês) ESTHER CAMARGO DE FONSECA 19h30 – 20h30 “Strategies of the 21stCentury Toxicology for a Safer and More Sustainable World” (Centro de Eventos da UFG) Dr. Thomas Hartung e Johns Hopkins University, EUA

20h30 – 23h COQUETEL DE ABERTURA Centro de Eventos da UFG PROGRAMAÇÃO OFICIAL

09 DE OUTUBRO DE 2017 – SEGUNDA-FEIRA HORÁRIO ATIVIDADE TEMA PALESTRANTE (INSTITUIÇÃO) SALA

8h – 18h CREDENCIAMENTO E ENTREGA DE MATERIAL SECRETARIA DO EVENTO (Centro de Eventos da UFG TOXICITY TESTING IN THE 21st CENTURY ESTER DE CAMARGO FONSECA MORAES 8h30 – 10h30 SIMPÓSIO 01 (em inglês) Chair: Dra. Marize Campos Valadares - Universidade Federal de Goiás (UFG) (Centro de Eventos da UFG) 8h30 – 9h15 S 1.1 Read-across Dr. Thomas Hartung - Johns Hopkins University, EUA

Dr. Troy Seidle 9h20 – 9h50 S 1.2 Toward a human pathway paradigm in safety testing and health research Human Toxicology Project Consortium (HTPC- Humane Society International), EUA

9h55 – 10:30h S1.3 Ocular Toxicity Dra. Chantra Eskes - ESTIV / SeCAM, Suíça NOVAS DROGAS DE ABUSO: MARIA ELISA DE SIQUEIRA 8h30 – 10h30 MESA-REDONDA 01 DA INTERNET PARA A SALA DE EMERGÊNCIA (Centro de Eventos da UFG) Chair: Dr. Luiz Carlos da Cunha - Universidade Federal de Goiás (UFG) 8h30 – 9h15 MR 1.1 Manifestações Clínicas Inusitadas nas Intoxicações Agudas por Cocaína Ms. Sergio Emmanuele Graff - TOXICLIN – SP

9h15 – 9h45 MR 1.2 Novas Substâncias Psicoativas: Diagnóstico Clínico e Manejo Inicial na Emergência Carlos Augusto Mello da Silva - TOXIKONSULT – RS

9h45 – 10h15 MR 1.3 Desaos Laboratoriais na Identicação das Novas Substâncias Psicoativas Dr. Rafael Lanaro - CIATox-Campinas

10h15 – 10h30 MR 1.4 DISCUSSÃO VIGILÂNCIA EM SAÚDE AMBIENTAL SILVIA BERLANGA BARROS 8h30 – 10h30 MESA-REDONDA 02 Chair: Dra. Gisele Augusto Rodrigues de Oliveira (Centro de Eventos da UFG) Universidade Federal de Goiás (UFG) 8h30 – 8h55 MR 2.1 Ecotoxicologia, Vigilância Ambiental e Saúde Dr. Eduardo Cyrino de Oliveira-Filho EMBRAPA – DF

Desaos da vigilância em saúde ambiental: um olhar sobre a detecção e Dr. Rhaul de Oliveira 8h55– 9h20 MR 2.2 os efeitos dos contaminantes emergentes nas águas brasileiras Universidade de São Paulo (FCF/USP)

9h20 – 9h40 MR 2.3 O papel do Ibama no registro de agrotóxicos no Brasil Msc. Déborah Mendes Máximo Cardozo - IBAMA Msc. Natiela Beatriz de Oliveira 9h40 – 10h MR 2.4 Vigilância em Saúde Ambiental no âmbito do Ministério da Saúde Coordenação Geral de Vigilância em Saúde Ambiental – CGVAM/MS 10h – 10h30 MR 2.5 DISCUSSÃO Visitação a feira COFFEE - BREAK 10h30 – 11h SESSÃO DE PÔSTER: Toxicologia Ambiental e Ecotoxicologia / Toxicologia Social / Toxicologia Forense / Toxicologia Analítica / Nanotoxicologia / Outras

SIMPÓSIO 02 NANOTOXICOLOGY ESTER DE CAMARGO FONSECA MORAES 11h – 13h (em inglês) Chair: Dr. Bennard van Ravenzwaay - BASF (Centro de Eventos da UFG)

11h -11h25 S2.1 Focus on the interactions between carbon-based nanomaterials and the immune system Dr. Bengt Fadeel - Karolinska Institutet, Suécia Dr. Ary Correa Júnior 11h30 – 11h55 S2.2 Regulatory research in nanotechnology Universidade Federal de Minas Gerais (UFMG) Dr Joachim Sturve Ecotoxicology of nanoparticles 12h – 12h25 S2.3 University of Gothenburg, Suécia Dra. Juliana Cancino-Bernardi 12h30 – 13h S2.4 Nanotoxicology of gold nanorods: in vitro, in vivo and membrane models Universidade de São Paulo (USP) TESTES MICROBIANOS PARA AVALIAÇÃO DE GENOTOXICIDADE Dr. Marcelo de Pádula MARIA ELISA DE SIQUEIRA 11h – 11h35 PALESTRA 01 NO DESENVOLVIMENTO DE FÁRMACOS Universidade Federal do Rio de Janeiro (UFRJ) (Centro de Eventos da UFG) Chair: Dra. Danielle Palma de Oliveira - Universidade de São Paulo (FCFRP/USP) DRIED BLOOD SPOTS: POTENCIAIS E DESAFIOS EM MONITORAMENTO Dr. Rafael Linden MARIA ELISA DE SIQUEIRA 11h40 – 12h15 PALESTRA 02 TERAPÊUTICO DE FÁRMACOSChair: Universidade Feevale (Centro de Eventos da UFG) Dr. Luiz Carlos da Cunha - Universidade Federal de Goiás (UFG)

PALESTRA 03 ASSESSMENT OF SKIN SENSITIZATION Msc. Allison Hilberer MARIA ELISA DE SIQUEIRA 12h20 – 13h (em inglês) Chair: Dra. Marize Campos Valadares - Universidade Federal de Goiás (UFG) Institute For In Vitro Sciences (IIVS, EUA) (Centro de Eventos da UFG)

POLÍTICAS PÚBLICAS SOBRE DROGAS NO BRASIL: UMA DISCUSSÃO EM TODOS OS NÍVEIS SILVIA BERLANGA BARROS 11h – 13h MESA-REDONDA 03 Chair: Dra. Camila Caixeta - Universidade Federal de Goiás (UFG) (Centro de Eventos da UFG) Coordenadora do Centro de Referências sobre Drogas da UFG (CRR/UFG)

Dra. Bárbara Vaz 11h – 11h30 MR 3.1 Políticas públicas sobre drogas: o panorama nacional Ministério da Saúde/FIOCRUZ

Msc. Fernanda Costa Nunes 11h30 – 12h MR 3.2 Políticas públicas sobre drogas: o panorama estadual Secretaria de Saúde do Estado de Goiás (SES - GO) / Universidade Federal de Goiás (UFG) Esp. Sheila Alves da Cunha 12h – 12h30 MR 3.3 Políticas públicas sobre drogas: o panorama local Secretaria Municipal de Saúde de Goiânia

12h30 – 13h DISCUSSÃO

Visitação a feira

ALMOÇO

I ENCONTRO DE LIGAS EM TOXICOLOGIA

Chair: Dra. Gisele Augusto Rodrigues de Oliveira - Universidade Federal de Goiás (UFG) MARIA ELISA DE SIQUEIRA 13h – 14h30 Apresentação das Ligas Acadêmicas de Toxicologia (Centro de Eventos da UFG) Coordenador/moderador: MSc. Éric Diego Barioni - Universidade de Sorocaba (UNISO)

Liga Acadêmica da UNINOVE (estudantes de medicina da UNINOVE) LATox-UFG (estudantes de farmácia da UFG)

Palestrante convidada: Dra. Silvia Cazenave “ Perspectivas da Toxicologia na área da Saúde”

MESA-REDONDA 04 REPRODUCTIVE AND DEVELOPMENTAL TOXICOLOGY ESTER DE CAMARGO FONSECA MORAES 14h30 - 16h30 (em inglês) Chair: Dra. Cibele Borges - Universidade Estadual Paulista (UNESP) (Centro de Eventos da UFG

Dr. Fernando Barbosa Junior 14h30 – 14h35 MR 4.1 Introduction Universidade de São Paulo (FCFRP/USP)

Dr. Warren Foster Brain Derived Neurotrophic Factor (BNDF) in the Diagnosis of Endometriosis 14h35 – 14h55 MR 4.2 McMaster University - Health Sciences Centre, Canadá

Dr. Thomas Knudsen 14h55 – 15h15 MR 4.3 Computational Modeling of Developmental Toxicity EPA, EUA

Dr. Daniel Cyr Gender-specic hepatic cancer induction by hexachlorobenzene: MR 4.4 Institut national de la recherche scientique 15h15 – 15h35 understanding mechanisms of toxicity. (INRS, Canadá)

Dra. Renata Mazaro Costa 15h35 – 15h55 MR 4.5 Male reproductive effects of Cannabidiol chronic treatment Universidade Federal de Goiás (UFG)

Prenatal betamethasone exposure causes intergenerational Dra. Cibele Borges 15h55 – 16h15 MR 4.6 developmental reprogramming in rats Universidade Estadual Paulista (UNESP)

16h15 – 16h30 MR 4.7 DISCUSSÃO

NOVAS SUBSTÂNCIAS PSICOATIVAS: OS DESAFIOS ANALÍTICO, REGULATÓRIO E FORENSE SILVIA BERLANGA BARROS 14h30 - 16h30 SIMPÓSIO 03 Chair: Dr. Maurício Yonamine - Universidade de São Paulo (FCF/USP) (Centro de Eventos da UFG)

Dr. José Luiz da Costa 14h30 – 15h10 S3.1 Novas Substâncias Psicoativas - aspectos analíticos Universidade Estadual de Campinas (UNICAMP) Dr. Rita Jorge 15h15 – 16h S3.2 Novas Substâncias Psicoativas – aspectos regulatórios European Monitoring Centre for Drugs and Drug Addiction (EMCDDA, Portugal)

Dr. Luciano Arantes 16h05 - 1630 S3.3 Novas Substâncias Psicoativas – aspectos forenses Perito Criminal da Polícia Civil do Distrito Federal

A CONSTRUÇÃO DO PERFIL DE SEGURANÇA DE VACINAS Msc. Beatriz da Costa Thomé MARIA ELISA DE SIQUEIRA 14h30 – 15h05 PALESTRA 04 Chair: Dra. Marize Campos Valadares - Universidade Federal de Goiás (UFG) Instituto Butantan - SP (Centro de Eventos da UFG)

O USO DE METODOLOGIAS ALTERNATIVAS NO CONTROLE DA QUALIDADE Dra. Isabella Fernandes Delgado MARIA ELISA DE SIQUEIRA 15h10 – 15h45 PALESTRA 05 DE VACINAS E SOROS HIPERIMUNES FIOCRUZ – RJ (Centro de Eventos da UFG) Chair: Dra. Marize Campos Valadares - Universidade Federal de Goiás (UFG)

PALESTRA 06 IN VITRO MODELS FOR RESPIRATORY TOXICOLOGY Dr. Rodger Curren MARIA ELISA DE SIQUEIRA 15h50 – 16h30 (em inglês) Chair: Dra.Marize Campos Valadares - Universidade Federal de Goiás (UFG) Institute For In Vitro Sciences (IIVS, EUA) (Centro de Eventos da UFG)

Visitação a feira

COFFEE - BREAK 16h30 – 17h SESSÃO DE PÔSTER: Toxicologia Ambiental e Ecotoxicologia / Toxicologia Social / Toxicologia Forense / Toxicologia Analítica / Nanotoxicologia / Outras

SESSÃO ORAL 01: ESTER DE CAMARGO FONSECA MORAES 17h – 18h15 Toxicologia Ambiental e Ecotoxicologia (Centro de Eventos da UFG)

SESSÃO ORAL 02 MARIA ELISA DE SIQUEIRA 17h – 18h15 Toxicologia Social; Toxicologia Forense (Centro de Eventos da UFG)

SESSÃO ORAL 03 SILVIA BERLANGA BARROS 17h – 18h15 Toxicologia Analítica; Nanotoxicologia; Outras (Centro de Eventos da UFG) ATIVIDADE SOCIAL / CULTURAL ESTER DE CAMARGO FONSECA MORAES 18h15 – 19h “Coletivo Desencuca” e “Banda Coró-de-Pau” (Centro de Eventos da UFG) Grupos de batucada que promovem ações relativas à saúde mental e à reintegração social de pessoas desassistidas, especialmente jovens.

19h HAPPY HOUR com Grupo Heróis de Botequim PROGRAMAÇÃO OFICIAL

10 DE OUTUBRO DE 2017 – TERÇA-FEIRA HORÁRIO ATIVIDADE TEMA PALESTRANTE (INSTITUIÇÃO) SALA SECRETARIA DO EVENTO 8h – 18h CREDENCIAMENTO E ENTREGA DE MATERIAL (Centro de Eventos da UFG) SIMPÓSIO 01 RISK ASSESSMENT IN THE 21st CENTURY ESTER CAMARGO DE FONSECA 8h30 – 10h30 (em inglês) Chair: Dra. Silvia Berlanga de Moraes Barros - Universidade de São Paulo (Centro de Eventos da UFG) Dra. Silvia Berlanga de Moraes Barros 8h30 – 9h S 1.1 Challenges risk assessment in the 21st century Universidade de São Paulo (FCF/USP)

9h05 – 9h30 S 1.2 In vitro models of dermal penetration in the pesticides risk assessment process Dr. Christian Strupp - ADAMA, Germany Incorporation of Knowledge of Integrating Toxicokinetics into Human Health 9h35 – 10h05 S 1.3 Dr. Manoj Aggarwal - Dow agrosciences USA Risk Assessment: A Proposed Framework and Case Studies

TOXICOLOGIA SOCIAL: VELHOS TOXICANTES OU NOVOS FÁRMACOS? MARIA ELISA DE SIQUEIRA 8h30 – 10h30 MESA-REDONDA 01 Chair: Dr. Luiz Carlos da Cunha - Universidade Federal de Goiás (UFG) (Centro de Eventos da UFG) Dra. Virgínia Martins Carvalho 8h30 – 9h MR 1.1 Perspectivas do uso da cannabis medicinal no Brasil: Projeto Farmacannabis Universidade Federal do Rio de Janeiro (UFRJ) Tratamento da dependência química com ibogaína e do estresse Dr. Eduardo Ekman Schenberg Universidade 9h – 9h30 MR 1.2 pós-traumático com MDMA Federal de São Paulo (UNIFESP) Dr. Rafael Guimarães dos Santos 9h30 – 10h MR 1.3 Propriedades terapêuticas da ayahuasca Universidade de São Paulo (FMRP/USP) 10h – 10h30 MR 1.5 DISCUSSÃO

IMMUNOTOXICOLOGY SILVIA BERLANGA BARROS 8h30 – 10h30 SIMPÓSIO 02 Chair: Daniel Junqueira Dorta - Universidade de São Paulo (FFCLRP/USP) (Centro de Eventos da UFG) Dra Sandra Farsky 8h30 – 9h15 S 2.1 Role of dioxins type compounds on immune system Universidade de São Paulo (USP Dr. Eduardo Lani Volpe da Silveira 9h20 – 9h50 S 2.2 Effects of cigarrette smoke compounds on immunization Universidade de São Paulo (USP) Dra. Silvana Sandri 9h55 – 10:30h S 2.3 Immune system on cancer development - Insights of mechanisms of nanocompounds Universidade de São Paulo (USP) Visitação a feira COFFEE - BREAK 10h30 – 11h SESSÃO DE PÔSTER: Imunotoxicologia / Toxinologia / Métodos Alternativos ao Uso de Animais / Toxicologia Ocupacional / Toxicologia de Alimentos

SIMPÓSIO 03 PERSPECTIVES OF METABOLOMICS IN TOXICOLOGY ESTER CAMARGO DE FONSECA 11h – 13h (em inglês) Chair: Dr. Márcio Lorencini - Grupo Boticário (Centro de Eventos da UFG) Dr. Bennard van Ravenzwaay 11h -12h S 3.1 The application of in vivo and in vitro metabolomics in toxicology BASF, Alemanha Dr. Ernani Pinto Junior 12h – 13h S 3.2 Advances in metabolomic and toxicology Universidade de São Paulo (FCF/USP)

GLOBAL LANDSCAPE OF GLYPHOSATE SAFETY EVALUATIONS Dra. Donna Framer MARIA ELISA DE SIQUEIRA 11h – 11h35 PALESTRA 01 Chair: Dr. Fernando Barbosa Junior - Universidade de São Paulo (FCFRP/USP) MONSANTO (Centro de Eventos da UFG)

PRODUÇÃO DE MATERIAL DE REFERÊNCIA CERTIFICADO DE DROGAS Dr. Rodrigo Borges de Oliveira SILVIA BERLANGA BARROS 11h – 11h35 PALESTRA 02 DE ABUSO E SEUS METABÓLITOS INMETRO – RJ (Centro de Eventos da UFG) Chair: Dr. Luiz Carlos da Cunha Universidade Federal de Goiás (UFG)

GLIFOSATO, PERIGO, RISCO E EXPOSIÇÃO HUMANA Dr. Francisco Paumgartten MARIA ELISA DE SIQUEIRA 11h40 – 12h15 PALESTRA 03 Chair: Dr. Fernando Barbosa Junior - Universidade de São Paulo (FCFRP/USP) FIOCRUZ (Centro de Eventos da UFG)

O LABORATÓRIO BRASILEIRO DE CONTROLE DE DOPAGEM E A EXPERIÊNCIA Dr. Henrique Marcelo Gualberto Pereira OLÍMPICA: AS ANÁLISES ANTIDOPAGEM DOS JOGOS OLÍMPICOS E SILVIA BERLANGA BARROS 11h40 – 12h15 PALESTRA 04 Laboratório Brasileiro de Controle de (Centro de Eventos da UFG) PARALÍMPICOS RIO 2016 Dopagem (LBCD - RJ) Chair: Dr. Luiz Carlos da Cunha - Universidade Federal de Goiás (UFG)

TOXINAS: DO VENENO AO MEDICAMENTO Dra. Marta Regina Magalhães MARIA ELISA DE SIQUEIRA 12h20 – 13h PALESTRA 05 Chair: Prof. MSc. Frank Sousa Castro Pontifícia Universidade Católica de Goiás (PUC - GO) (Centro de Eventos da UFG) Pontifícia Universidade Católica de Goiás (PUC - GO) ANÁLISE DE THC E METABÓLITOS EM CABELO PELA TÉCNICA DE GC-MSMS Romão Beserra Jr SILVIA BERLANGA BARROS 12h20 – 13h PALESTRA 06 Chair: Dr. Tiago Franco de Oliveira Especialista de Produto Agilent (Centro de Eventos da UFG) Universidade Federal de Ciências da Saúde de Porto Alegre (UFSCPA)

Visitação a feira

ALMOÇO

I ENCONTRO DE LIGAS EM TOXICOLOGIA Liga Acadêmica da UNINOVE (estudantes de medicina da UNINOVE) MARIA ELISA DE SIQUEIRA 13h – 14h30 LATox-UFG (estudantes de farmácia da UFG) (Centro de Eventos da UFG) Chair: Dra. Gisele Augusto Rodrigues de Oliveira Universidade Federal de Goiás (UFG)

Dra. Ingrid Dragan Taricano Universidade Nove de Julho (UNINOVE)

MODELS AND PRACTICES TO EVALUATE THE OCCUPATIONAL RISK TO MESA-REDONDA 02 ESTER CAMARGO DE FONSECA 14h30 - 16h30 PESTICIDES HANDLERS - BRAZILIAN AND INTERNATIONAL SCENARIOS (em inglês) Chair: Dra. Silvia Berlanga de Moraes Barros (Centro de Eventos da UFG) Universidade de São Paulo (FCF/USP) Presentation of the international occupational exposure databases Dr. Victor Cañez 14h30 – 15h15 MR 4.1 (AHED, PHED and EFSA) Risk Assessment Solutions, EUA Fabiana C. Palma 15h15 – 16h MR 4.2 ProHuma initiatives to support the assessment of occupational risk in Brazil ProHuma

16h – 16h30 DISCUSSÃO

MÉTODOS ALTERNATIVOS - ASPECTOS REGULATÓRIOS, CIENTÍFICOS E ACEITAÇÃO MARIA ELISA DE SIQUEIRA 14h30 - 16h30 Chair: Marize Campos Valadares SIMPÓSIO 04 (Centro de Eventos da UFG) Universidade Federal de Goiás (UFG)

Dr. Octavio Presgrave 14h30 – 14h55 S 4.1 Métodos alternativos - aspectos regulatórios FIOCRUZ - RJ

Dra. Chantra Eskes S 4.2 Métodos alternativos – aceitação 15h – 15h25 ESTIV / SeCAM, Suíça

Cultivo simultâneo de vários tecidos humanos em chips microuidicos Dr. Eduardo Pagani 15h30 – 15h55 S 4.3 para testes toxicológicos in vitro Laboratório Nacional de Biociências (LNBio - SP) Aplicação de métodos alternativos para avaliação da ecácia e Dr. Márcio Lorencini 16h – 16h30 S 4.4 segurança de cosméticos Grupo Boticário - PR

GENÔMICA NUTRICIONAL Dra. Thais Steemburgo SILVIA BERLANGA BARROS 14h30 – 15h05 PALESTRA 07 Chair: Dra. Danielle Palma de Oliveira - Universidade de São Paulo (FCFRP/USP) Universidade Federal do Rio Grande do Sul (UFRGS) (Centro de Eventos da UFG)

AUSÊNCIA DE COMBUSTÃO NO USO DE TABACO REDUZ O RISCO TOXICOLÓGICO?

Chair: Dra. Danielle Palma de Oliveira Universidade de São Paulo (FCFRP/USP)

Mediadora: Dra. Silvia Cazenave Instituto de Criminalística de Campinas SILVIA BERLANGA BARROS 15h10 – 16h30 DEBATE (Centro de Eventos da UFG) Debatedores: Dr. Marcus da Matta Lisam Ecoadvisor Systems Dr. Paulo César Rodrigues Pinto Corrêa Universidade Federal de Ouro Preto (UFOP) Membro da Comissão de Tabagismo (SBPT)

Visitação a feira

COFFEE - BREAK 16h30 – 17h SESSÃO DE PÔSTER: Imunotoxicologia / Toxinologia / Métodos Alternativos ao Uso de Animais / Toxicologia Ocupacional / Toxicologia de Alimentos

SESSÃO ORAL 04: ESTER CAMARGO DE FONSECA 17h – 18h15 Métodos Alternativos ao Uso de Animais (Centro de Eventos da UFG) SESSÃO ORAL 05: MARIA ELISA DE SIQUEIRA 17h – 18h Imunotoxicologia; Toxinologia (Centro de Eventos da UFG) SESSÃO ORAL 06: SILVIA BERLANGA BARROS 17h – 18h15 Toxicologia Ocupacional; Toxicologia de Alimentos (Centro de Eventos da UFG) MARIA ELISA DE SIQUEIRA 18h – 19h30 ASSEMBLEIA GERAL DA SOCIEDADE BRASILEIRA DE TOXICOLOGIA (SBTox) (Centro de Eventos da UFG)

19h30 – 22h HAPPY HOUR com Rodrigo Arantes PROGRAMAÇÃO OFICIAL

11 DE OUTUBRO DE 2017 – QUARTA-FEIRA

HORÁRIO ATIVIDADE TEMA PALESTRANTE (INSTITUIÇÃO) SALA

PRODUTOS DE DEGRADAÇÃO DE FÁRMACOS: ASPECTOS REGULATÓRIOS E DESAFIOS ANALÍTICOS ESTER DE CAMARGO FONSECA MORAES 8h – 10h MESA-REDONDA 01 Chair: Dr. Luiz Carlos Cunha Universidade Federal de Goiás (Centro de Eventos da UFG)

Dr. Rogério Hoeer 8h – 8h30 MR 1.1 Produtos de degradação de fármacos: aspectos regulatórios Conselho Federal de Farmácia – CEBRIM Dr. Leonardo de Souza Teixeira 8h30 – 9h MR 1.2 Produtos de degradação de fármacos: desaos analíticos ICF - GO Dr. Helio Martins 9h – 9h30h MR 1.3 Produtos de degradação de fármacos: avanços analíticos SCIEX do Brasil

9h30 – 10h MR 1.4 DISCUSSÃO

SIMPÓSIO 01 ENDOCRINE DISRUPTORS MARIA ELISA DE SIQUEIRA 8h – 10h (em inglês) Chair: Dr. Rhaul de Oliveira - Universidade de São Paulo (FCF/USP) (Centro de Eventos da UFG)

Dr. Gustavo Somoza Effect s of waterborne xenoestrogens on feminization of the testicular structure 8h – 8h50 S 1.1 Universidad Nacional de General San Martín of two neotropical sh. (UNSAM), Argentina

Dra. Helena C. da Silva de Assis 9h – 10h S 1.2 Effects of endocrine disruptors at low concentration Universidade Federal do Paraná (UFPR)

COMPOSTOS TÓXICOS FORMADOS DURANTE O PROCESSAMENTO Dra. Alessandra Vincenzi Jager SILVIA BERLANGA BARROS 8h – 8h35 PALESTRA 01 TÉRMICO DOS ALIMENTOS Universidade de São Paulo (FCFRP/USP) (Centro de Eventos da UFG) Chair: Dra. Elisa R. Anastácio Ferraz - Universidade Federal Fluminense (UFF)

HARMONIZAÇÃO E DISSEMINAÇÃO DE METODOLOGIAS DE TESTE: ACEITAÇÃO MÚTUA DE DADOS E BOAS PRÁTICAS Dra. Luciene B Balottin SILVIA BERLANGA BARROS 8h40 – 9h15 PALESTRA 02 Chair: Dra. Elisa R. Anastácio Ferraz IMETRO (Centro de Eventos da UFG) Universidade Federal Fluminense (UFF)

MODELOS DE NEUROTOXICIDADE DO VÍRUS ZIKA DURANTE O Dra. Patrícia Pestana Garcez SILVIA BERLANGA BARROS 9h20 – 10h PALESTRA 03 DESENVOLVIMENTO DO CÓRTEX CEREBRAL Universidade Federal do Rio de Janeiro (Centro de Eventos da UFG) Chair: Dra. Elisa R. Anastácio Ferraz - Universidade Federal Fluminense (UFF) (UFRJ)

Visitação a feira

COFFEE - BREAK 10h – 10h30 SESSÃO DE PÔSTER: Toxicologia Experimental / Genômica, Proteômica e Metabolômica / Mutagênese e Carcinogênese / Toxicologia Clínica / Toxicologia de Medicamentos e Cosméticos / Toxicologia Veterinária

ESTUDOS PRÉ-CLÍNICOS E PESQUISA CLÍNICA APLICADOS À I ESTER DE CAMARGO FONSECA MORAES 10h30 – 12h30 MESA-REDONDA 02 NOVAÇÃO FARMACÊUTICA (Centro de Eventos da UFG) Chair: Dra. Renata Mazaro Costa - Universidade Federal de Goiás

Msc. Ednilza Pereira de Farias Dias 10h30 – 11h05 MR 2.1 Aspectos Éticos da Pesquisa Clínica, uxo CEP/CONEP PPD do Brasil - Suporte à Pesquisa Clínica Dra. Ingrid Dragan Taricano 11h05 – 11h40 MR 2.2 Toxicologia não-clínica no desenvolvimento de novos fármacos Universidade Nove de Julho (UNINOVE) Msc. Adriane Alves de Oliveira 11h40 – 12h10 MR 2.3 Ensaios pré-clínicos para aplicados à inovação farmacêutica ANVISA

12h10 – 12h30 MR 2.4 DISCUSSÃO TOXICOLOGIA E A GESTÃO SEGURA DE SUBSTÂNCIAS QUÍMICAS NO BRASIL SILVIA BERLANGA BARROS 10h30 – 12h30 MESA-REDONDA 03 Chair: Dra. Flávia Neri Meira de Oliveira - Secretaria da Saúde do Distrito Federal (Centro de Eventos da UFG)

Msc. Fabriciano Pinheiro 10h30 – 11h MR 3.1 Toxicologia como base da Segurança Química INTERTOX – SP Msc. Camila Arruda Boechat Ministério 11h – 11h30 MR 3.2 Visão geral sobre a legislação de controle de substâncias químicas no Brasil do Meio Ambiente - DF Ações da Indústria Química Brasileira para promover a gestão Msc. Fernando Correia de Moraes Tibau MR 3.3 11h30 – 12h segura de substâncias químicas Associação Brasileira da Indústria Química (ABIQUIM)

12h – 12h30 DISCUSSÃO

INOVAÇÕES NO TESTE DE SALMONELLA/MICROSSOMA Dra. Gisela A. Umbuzeiro MARIA ELISA DE SIQUEIRA 10h30 – 11h10 PALESTRA 01 Chair: Dra. Danielle Palma de Oliveira Universidade Estadual de Campinas (Centro de Eventos da UFG) Universidade de São Paulo (FCFRP/USP) (UNICAMP)

EPIGENÉTICA E TOXICOLOGIA: ALTERAÇÕES EPIGENÉTICAS RELACIONADAS À EXPOSIÇÃO A AGENTES TÓXICOS Dr. Ramon Bossardi Ramos MARIA ELISA DE SIQUEIRA 11h10 – 11h50 PALESTRA 02 Chair: Dra. Gisela A. Umbuzeiro - Universidade Estadual de Campinas Universidade Anhembi Morumbi (Centro de Eventos da UFG) (UNICAMP)

TOXICOLOGIA OCUPACIONAL: BIOMARCADORES NOVOS, TRADICIONAIS E Dr. José Salvador Lepera MARIA ELISA DE SIQUEIRA 11h50 – 12h30 PALESTRA 03 A REGULAMENTAÇÃO NO BRASIL Universidade Estadual Paulista (UNESP) (Centro de Eventos da UFG) Chair: Dra. Fabiana Cremaschi Palma - ProHuma

Visitação a feira 12h30 – 14h ALMOÇO

MÉTODOS ALTERNATIVOS PARA AVALIAÇÃO DE CONTAMINANTES EMERGENTES SILVIA BERLANGA BARROS 14h – 15h20 MESA-REDONDA 04 Chair: Dra. Gisele A. Rodrigues de Oliveira (Centro de Eventos da UFG) Universidade Federal de Goiás Dra. Danielle Palma de Oliveira 14h – 14h25 MR 4.1 Contaminantes emergentes – resultados com zebrash Universidade de São Paulo (FCFRP/USP) Dra. Elisa R. Anastásio Ferraz Avelino 14h25 – 14h45 MR 4.2 Cultura 3D em hidrogel de alginato para avaliação de corantes têxteis Universidade Federal Fluminense (UFF) Dr. Daniel Junqueira Dorta 14h45 – 15h05 MR 4.3 Avaliação da toxicidade de retardantes de chamas bromados utilizando métodos in vitro Universidade de São Paulo (FFCLRP/USP)

15h05 – 15h20 DISCUSSÃO

30 ANOS DO ACIDENTE COM CÉSIO E A TOXICIDADE DE OUTROS ESTER DE CAMARGO FONSECA MORAES 14h - 16h SIMPÓSIO 02 ELEMENTOS RADIOATIVOS (Centro de Eventos da UFG Chair: Prof. Dr. Luiz Carlos da Cunha - Universidade Federal de Goiás Dra. Adriana Costa Guimarães 14h – 14h25 S 2.1 Elementos radioativos: um enfoque nos estudos da radiação com urânio Universidade Federal do Pará - UFPA Dr. Alonso Monteiro da Silva S 2.2 Aspectos clínicos da população exposta ao césio 14h30 – 14h55 Centro de Informação Toxicológica de Goiás (CIT - GO) Médico Paulo José Malheiros Fialho 15h – 15h25 S 2.3 Passado, presente e futura na assistência aos radioacidenados Centro de Assistência aos Radioacidentados - CARA Dr. Aparecido Divino da Cruz 15h30 – 16h S 2.4 Retrospectiva dos efeitos genotóxicos do césio sobre a população exposta Pontifícia Universidade Católica de Goiás - PUC - GO

Seleção de endpoint toxicológico e fatores de incerteza para Avaliação do RiscoAvaliação do Consumo Agudo e Crônico de Alimentos In Natura da População Brasileira por Meio da Pesquisa de Orçamentos Familiares - Dra. Giovanna Semino-Beninel Bayer MARIA ELISA DE SIQUEIRA 14h – 15h20 PALESTRA 04 Bloco de Consumo Alimentar Pessoal (IBGE POF 2008/2009), para realização Cristiana Corrêa (CCA ILSI Brasil) (Centro de Eventos da UFG) da avaliação de exposição a resíduos de agrotóxicos Cristiana Corrêa (CCA ILSI Brasil)

EXPOSIÇÃO AO ARSÊNIO PELO CONSUMO DE ARROZ NO BRASIL: DEVEMOS Dr. Fernando Barbosa Junior SILVIA BERLANGA BARROS 15h20 – 16h PALESTRA 05 NOS PREOCUPAR? Universidade de São Paulo (FCFRP/USP) (Centro de Eventos da UFG) Chair: Gustavo Somoza - Universidad Nacional de General San Martín (UNSAM)

SESSÕES ORAIS: ESTER DE CAMARGO FONSECA MORAES 16h - 17h Toxicologia Experimental (Centro de Eventos da UFG SESSÕES ORAIS: MARIA ELISA DE SIQUEIRA 16h – 17h Genômica, Proteômica e Metabolômica; Mutagênese e Carcinogênese (Centro de Eventos da UFG) SESSÕES ORAIS: SILVIA BERLANGA BARROS 16h – 17h Toxicologia Clínica; Toxicologia de Medicamentos e Cosméticos; Toxicologia Veterinária (Centro de Eventos da UFG)

PREMIAÇÃO DOS MELHORES TRABALHOS (ORAL E PÔSTER) ESTER DE CAMARGO FONSECA MORAES 17h – 18h (Centro de Eventos da UFG ENCERRAMENTO

ALTERNATIVE METHODS TO MAMMALIAN MODELS ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 18

AM 01 - INFLAMMATORY PROFILE OF AM 02 - HUMAN ADIPOSE-DERIVED STEM HUMAN KERATINOCYTES AS A NON- CELLS AND HUMAN PERIODONTAL ANIMAL METHOD FOR OCULAR TOXICITY LIGAMENT STEM CELLS AS CELLULAR EVALUATION SUBSTRATES OF A TOXICITY PREDICTION ASSAY CHIALCHIA, A.R. 1; SILVA, A.C.G. 1; VALADARES, M.C. 1 Aguiar, A.M.b, Corrêa, NCRa, Kuligovski, Cb, ACCb, Abud, APRAb, Rebelatto, CLKc, Leite, LMBc, 1 Laboratório de Farmacologia e Toxicologia Celular, Senegaglia, ACc, Dallagiovanna, Bb and Aguiar, AMb Faculdade de Farmácia, Universidade Federal de Goiás, Goiânia-GO, Brasil. aLaboratório de Nanobiotecnologia, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil. Introduction: Eye toxicity is a mandatory toxicological bLaboratório de Biologia Básica de Células-tronco, parameter for the classification and labelling. The Instituto Carlos Chagas, Fiocruz, Curitiba, PR, Brazil. Draize rabbit eye test has been used for assess eye cNúcleo de Tecnologia Celular (NTC), Pontifícia irritation and corrosion is, which consists in a in vivo Universidade Católica do Paraná (PUC-PR), Curitiba, subjective evaluation and does not allow the PR, Brazil. mechanistic evaluation of eye toxicity process. Furthermore, this in vivo method was never formally Introduction With the increasing need to develop in validated and although several alternative methods are vitro assays to replace animal use, human stem cell- available to eye irritation assessment, none of them derived methods are emerging and showing integrates inflammatory endpoints in their evaluated outstanding contributions to the toxicological parameters. Previous studies from our group screening of substances. Objective In this work, adult demonstrated that human keratinocytes displays human stem cells such as adipose-derived stem cells alterations in cytokine production profile in response (ADSC) and periodontal ligament stem cells (PDLSC) to exposure to eye irritants. Objective: In this study, were used as cell substrates for a cytotoxicity assay production of pro-inflammatory biomarker IL-1β was and toxicity prediction using the neutral red uptake evaluated after exposure of HaCat keratinocytes cells (NRU) assay. Materials Methods Adipose-derived to eye irritants and non-irritants substances, aiming to stem cells (ADSC) and periodontal ligament stem cells stablish a correlation between inflammation and eye (PDLSC) from 3 independent donors were toxicity. Material and Methods: As a first step, 12 characterized as mesenchymal stem cells (MSC) and chemicals were select including 3 non-irritants and 3 then these cell substrates were evaluated in NRU assay irritants from each Globally Harmonized System eye for cytotoxicity. Results First, ADSC and PDLSC were irritation category (Category 1, 2A and 2B). characterized as mesenchymal stem cells (MSC) by Posteriorly, HaCat were cultivated and exposed for 24 plastic adherence and appropriate immunophenotypic hours to the Cell Viability 80% concentration (CV80) MSC markers (positive for CD90, CD73, and CD105 of each chemical, obtained previously using MTT and negative for CD11b, CD34, CD45, HLADR, and cytotoxicity assay. Finally, IL-1β quantification in cell CD19). Furthermore, ADSC and PDLSC were able to lysates was carried out using a BD Opteia® ELISA kit, differentiate into adipocytes and osteoblasts when according to manufacturer’s instructions. Results: maintained under the same culture conditions Outcomes suggests that IL-1β production is altered by previously established for the NRU assay. NRU assays eye irritants exposure of HaCat cells. Mild, moderate for three reference test substances were performed. R2 and severe irritants induced a gradual increase in this was higher than 0.85 for all conditions, showing the cytokine production, while non-irritants showed levels feasibility to calculate IC50 values. The IC50 values similar to negative control. Conclusion: The findings were then used to predict the LD50 of the test show that IL-1β production by human keratinocytes substances, which were comparable to previous results demonstrated a good correlation with GHS eye and the ICCVAM standard test report. Conclusion irritation classification of the chemicals tested allowing Primary ADSC and PDLSC showed the potential to be the discrimination between eye irritants and non- considered as additional models for use in cytotoxicity irritants substances. assays. Financial support/Acknowledgments: CAPES, Financial support/Acknowledgments: CNPq, CNPQ, FUNAPE, FINEP. Fundação Araucária, CAPES, FIOCRUZ, Plataformas tecnológicas da FIOCRUZ.

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 19

AM 03 - ASSESMENT OF A NEW AM 04 - INIBITION OF ADULT STEM CELLS RECONSTRUCTED HUMAN EPIDERMIS DIFFERENTIATION FOR TOXICITY MODEL BY SKIN ABSORPTION TEST PREDICTION AS AN ALTERNATIVE METHOD TO ANIMAL TESTING AGUILLERA-FORTE, A. L. S. 1; TANGERINO, F. F 1,2; MASSARIOLI, B. 1,2; FONSECA, M. J. V. 2; ABUD, A.P.R.*1; KULIGOVSKI, C.1,5; CORREA, MARQUELE-OLIVEIRA, F. 1 N.C.R.2; MORAES, E.3; REUS, T.L.1,4; PASCHOAL, A.C.C.1; CARUSO, R.1,5; SCHUCK, 1 ELEVE Research and Development, Ribeirão Preto - D.C.6; BROHEM, C.A.6; MUÑIZ, B.D.1; AGUIAR, SP, Brazil; 2 University of Sao Paulo – Faculty of A.M.1 Pharmaceutical Science of Ribeirao Preto, Ribeirão Preto - SP, Brazil. 1 Laboratório de Biologia Básica de Células-tronco, Instituto Carlos Chagas, Fiocruz, Curitiba, PR. Brazil; Introduction: Information on skin toxicity and 2 Laboratório de Nanobiotecnologia, Universidade (trans)dermal penetration of compounds is of the Federal de Uberlândia, Uberlândia, MG. Brazil; 3 greatest relevance to guarantee the safety of consumer Universidade Federal do Rio de Janeiro. Rio de products. Reconstructed human epidermis (RHE) is Janeiro, RJ. Brazil; 4 Universidade Federal do Paraná, commonly employed for the purposes of regulatory Curitiba, PR. Brazil; 5 Instituto de Biologia Molecular toxicological testing, once it reproduces the do Paraná – IBMP, Curitiba PR. Brazil; 6 Grupo morphological, biochemical and physiological Boticário, Curitiba, PR. Brazil. properties of the human epidermis. Validation studies in Europe indicated that RHE commercial models are Introduction: Animal testing is a crucial way to verify suitable for in vitro absorption tests according to activity and security of substances. However, it has OECD TG 428. However, in Brazil, commercial been questioned worldwide resulting in several efforts models are not available yet, emphasizing the to reduce, replace or refine their use. There are many importance and the urgency of more studies in this alternative methods to the animal use but few with area. Aim: The aim of this study was to evaluate the human cells and even less using stem cells as a integrity of RHE developed by caffeine diffusion test. substrate. Previous studies have shown the ability of Methods: Human keratinocytes were seeded in culture adult stem cells to serve as a substrate for toxicity inserts and kept submerged in a 3:1 DMEM and HAM testing with similar, or even better, results than mixture with 10% fetal bovine serum and growth reference lineage (1). Human adipose tissue-derived factors during 2 (condition 1) or 7 days (condition 2). stem cells (hADSCs) are tissue resident stem cells Then, the inserts were raised and maintained at the air– responsible for tissue homeostasis and regeneration liquid interface for 13 (condition 1) or 15 days following injury. Stem cells are a good model for (condition 2). Diffusion studies were carried out using predicting toxicity because they can be used to static Franz Diffusion Cells with 12mL of phosphate evaluate cytotoxic effects on cell viability as well as buffer solution (pH 7.4) in the receptor compartment the cell differentiation process. Objectives: In this and 0.5 mL of caffeine solution (1 mg/mL) in the work, adipogenesis inhibition was assessed as an in donor compartment. Samples were collected for 8 vitro method for toxicity prediction of a test substance, hours and further analyzed by HPLC. Six replicates by using hADSCs during adipogenesis. Material and were used for each condition. Results: The results Methods: hADSCs from three patients were isolated, showed that different cultivation conditions were able cultured, characterized and maintained under to change epidermal diffusion of caffeine. RHE differentiation conditions during 14 days. In this study, cultivation under condition 1 presented the apparent 12 reference substances were used (2). During permeability coefficient (Paap) 15.40E-6±0.91E-6 adipogenesis in the presence of each one of those cm/s and permeation 198.30±6.17 µg/cm2 after 6 substances, we analyzed three different parameters hours, while condition 2 RHE presented Paap 7.9E- such as cell number, % of differentiated cells and area 6±0.52E-6 cm/s and permeation 148.96±11.10 of cell differentiation, to obtain IC50 values. Results: µg/cm2. Although these values are superior than those We could observe a dose-response curve, associated found to caffeine absorption in human skin, they are with the presence of the test substance during the comparable to other models reported in literature. adipogenesis. We also calculated the predicted the Conclusion: The RHE model developed using LD50 for the three parameters evaluated. Our results condition 2 might be a useful model to evaluate skin have shown that adipogenesis inhibition gives a more permeability of drugs, chemicals and cosmetics accurate class prediction to highest toxic substances compounds and to predict their toxicity. and incorrect prediction to lower toxic classes in Financial support/Acknowledgments: “Fundação de comparison to the current alternative in vitro toxicity Amparo à Pesquisa do Estado de São Paulo” - method. Some test substances have shown a FAPESP for financial support and research fellowship. stimulatory effect of adipogenesis, which can be ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 20 considered as another way to verify cytotoxicity. rate of false negative. Conclusion: the 3D model Conclusion: These results showed that our method constructed with keratinocytes can be considered a could be a complementary alternative assay to address valuable tool for assessing eye irritation potential of toxicity in vitro. mixtures, in particular, those considered that not Financial support / Acknowledgments: CAPES, require classification for eye irritation due its low Fundação Araucária, CNPq, Fiocruz. potential of toxicity, confirming the domain of applicability of the RhCE. Financial support/Acknowledgments: CAPES; AM 05 - RECONSTRUCTED HUMAN CORNEA- CNPQ; FINEP. LIKE EPITHELIUM TEST FOR OCULAR TOXICITY ASSESSMENT OF EYEBROW HENNA AM 06 - 3D EPITHELIAL MODEL FOR OCULAR TOXICITY EVALUATION: SILVA, A. C. G. 1; CHIALCHIA, A. R. 1; SANTOS, CHARACTERIZATION AND PROFICIENCY T. R. M. 1; ANDRADE, W. M. 1; BATISTA, A. C. 2; VALADARES, M. C 1 SILVA, A. C. G. 1; CHIALCHIA, A. R. 1; CASTRO, E. G. 1; ARANTES, D. A. C. 2; BATISTA, A. C. 2; 1 Laboratório de Farmacologia e Toxicologia Celular, VALADARES, M. C. 1. Universidade Federal de Goiás, Goiânia-GO, Brasil; 2 Laboratório de Patologia Oral, Faculdade de 1 Laboratório de Farmacologia e Toxicologia Celular, Odontologia, Universidade Federal de Goiás, Goiânia - Faculdade de Farmácia, Universidade Federal de Goiás GO, Brasil. - GO, Brasil; 2 Laboratório de Patologia Bucal, Faculdade de Odontologia, Universidade Federal de Introduction: Toxicological assessment of cosmetic Goiás, Goiânia-GO, Brasil. has been carried out using non-animal based tests since the use of animals was banished for this purpose. In Introduction: Ocular toxicity has been investigated this context, validation studies indicated Reconstructed through the Draize eye irritation test, using rabbits, for human Cornea-like Epithelium (RhCE) commercial more than 50 decades. However, ethical, economic and model as an in vitro assay appropriate for investigating scientific issues have made animal experimentation eye toxicity potential of products that do not require unsustainable. In vitro and ex vivo ocular toxicity classification for eye irritation (OECD 492), due to its testing includes cell and tissue-based methods that low rate of false negative. However, in Brazil, this enables classification of chemicals and mixtures commercial model is not available yet, showing the without animal testing. Among these, epithelial models importance of the development of 3D models like that. have been successfully employed to investigate eye Objective: In this study, we developed, characterized a irritation. Objective: In this study, we developed and 3D epithelial model and tested the applicability of the characterized a 3D stratified epithelial model, as well model in assess eye irritation potential of five eyebrow as investigated its proficiency as a method to assess henna samples, retailed as a non-irritant cosmetic eye toxicity potential of chemicals. Material and product. Methods: 3D epithelial models were obtained Methods: 3D epithelial models were obtained through through cultivation of HaCat human keratinocytes onto cultivation of HaCat human keratinocytes onto a Type a Type I collagen matrix for 5 days, at an air-liquid I collagen matrix for 5 days, at an air-liquid interface interface in cell culture inserts and tissue morphology in cell culture inserts. Expression of biomarkers (ki-67, was analyzed and characterized. Eyebrow henna Pan-cytokeratin and CD44) was performed to samples were tested as 10% dispersion in Phosphate- characterize it using immunohistochemistry and Buffer Saline (PBS), so that 50µL of suspension was immunofluorescence techniques. For testing the deposited onto the surface of epithelial models for 5 proficiency of epithelial model, 12 chemicals were minutes. Tissue viability was evaluated 24 hours after selected distributed in the Globally Harmonized exposure using MTT reduction assay. Short Time System eye irritation categories: Category 1, 2A and Exposure test (STE) (OECD 491, low rate of false 2B. Chemicals were tested by direct application onto positive) was used as a comparison method to assess epithelial surface of models for 5 minutes and tissue eye irritation potential of samples. Results: The new viability was assessed 24 hours after exposure using epithelial 3D model acquired a stratified morphology MTT cytotoxicity test. Results: Outcomes after cultivation and four eyebrow henna samples demonstrated that after 5 days of cultivation, HaCat decreased significantly tissue viability after exposure. cells formed a stratified epithelial structure (4-6 According to obtained data, three samples were layers), with regular expression of epithelial classified as irritants and two products as mild or non- biomarkers ki-67, Pan-cytokeratin and CD44. irritants, while STE did not classify most of the Furthermore, exposure to chemicals showed that samples. Its worth to mention that STE test has a high severe and moderated irritants promotes a high ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 21 decrease in tissue viability, followed by mild irritants, combinations can lead to ROS generation after UV while non-irritants did not altered significantly this exposure, which not necessarily translates to parameter. Conclusion: the 3D epithelial model phototoxic potential. Moreover, it was demonstrated constructed with HaCat human keratinocytes that bemotrizinol (5%) present in the combination C4 demonstrated a multilayered epithelial morphology provided a reduction of photoreactivity when suitable to investigate ocular toxicity and for compared to the combination C1, which did not classification of chemicals as ocular irritants and non- contain this UV filter. irritants. Financial support/Acknowledgments: CAPES, Financial support/Acknowledgments: CAPES, FAPESP (processo nº 2015/10017-3). CNPQ, FUNAPE, FINEP.

AM 08 - REAL-TIME PLATFORM FOR IN AM 07 - PHOTOTOXICITY PREDICTION OF SILICO TOXICOLOGICAL SCREENING AND UV FILTER COMBINATIONS USING ROS INTERFACE WITH VIRTUAL LABORATORY ASSAY AND 3T3 NRU PHOTOTOXICITY TEST AS AN ALTERNATIVE METHOD TO ANIMAL USING IN BRAZIL KAWAKAMI, C. M. 1; GASPAR, L. R. 1 SANTOS, C.E.M.1 1. Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP/USP) 1. Altox Ltda. São Paulo-SP, Brasil.

Efficient UV-absorbing molecules are designed to Introduction: Considering a growing need to assess the protect against UV-light exposure. However, the toxicity and ecotoxicity of a lot of chemicals (new development of photostable UV filter combinations is molecules, pharmaceuticals, cosmetic ingredients, important to guarantee the photoprotective efficacy additives, pesticides, impurities etc.) and the current and to prevent the formation of reactive oxygen trends of rationalization of animal use, alternative species (ROS), which can lead to phototoxic reactions methods like toxicological in silico has been on skin 1. The aim of this study was to evaluate the recommended in specific international guidelines and phototoxic potential of UV filter combinations with recently in Brazil. These models are considered useful, different photostability profiles by using ROS assay since the potential for toxicity and ecotoxicity are and 3T3 NRU phototoxicity test. For this purpose, four assessed by softwares, without the need of synthesis different UV filter combinations were prepared: C1 (obtaining the physical form) of the substance or (containing avobenzone, octyl methoxycinnamate) and biological materials. Incorporation of these in silico C3 (containing avobenzone, octyl methoxycinnamate, screening by Brazilian industries is still limited by 3% bemotrizinol), which were previously considered several factors: lack of local specialised knowledge, photounstable and C2 (DHHB and octyl complexity of computational modeling laboratory methoxycinnamate) and C4 (containing avobenzone, design, and divergence of specific focus areas of R&D octyl methoxycinnamate, 5% bemotrizinol), which (Research & development) that can result in technical were previously considered photostable. The ROS and/or economic infeasibility. Objective: To enable the assay was carried out for the detection of both singlet first Latin American (LA) platform of Computational oxygen and superoxide generation, according to Toxicology regulatory-based and to make accessible JACVAM (2013) 2 and OECD TG DRAFT 3. The the toxicological screening in silico (toxicity phototoxicity potential was evaluated by using 3T3 assessments by chemical structure) and in real time. monolayer fibroblast culture, which was submitted to Methodology: It is in development the optimization of UVA radiation for the determination of cell viability in a virtual lab with resources and adequate structure for the presence and absence of radiation, according to implementation of a realtime online platform. OECD TG 432 4. The results of ROS assay Quantitative structure–activity relationship (QSAR) demonstrated that combinations C1 and C3, which models for mutagenicity, acute toxicity, ADME were considered photounstable, were classified as endpoints, aquatic toxicity and environmental fate will photoreactive (SOC1: 42 and SA: -23; SOC3: 27.66 be incorporated based in regulatory guidelines. Once and SAC3: -18). On the other hand, combinations C2 deployed, team training will be held on the use of and C4, which are considered photostable, did not models and protocols for obtaining reports. Results and present any photoreactivity potential (SOC2: 4.33 and conclusion: it is expected that the process of in silico SAC2: -21.66; SOC4: 20.66 and SAC4: -22.66). The screening in remote mode by means of obtaining results of 3T3 NRU phototoxicity test showed that all (specialized team, structured lab and interaction UV filter combinations did not any present phototoxic platform) will result in positive impacts for R&D potential (MPE values between -0.081 and 0.072). The sector of new substances in Brazil and in LA, results showed that photounstable UV filter including costeffectiveness, rational use of animals and ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 22 selecting sustainable molecules, attraction of new AM 10 - IDENTIFICATION OF GENES THAT projects, and return on image in establish the first MODULATE SUSCEPTIBILITY TO DIPHENYL virtual laboratory of toxicology in real time in LA. DISELENIDE BY FUNCTIONAL TOXICOLOGY Financial support/Acknowledgments: FAPESP - IN YEAST Fundação de Amparo à Pesquisa do Estado de São Paulo. DALLA CORTE C.L. 1,2; OCCAI B.K. 1; REIS M.M. 3; MASUDA C.A. 4; LOMELI M.M. 4; ROCHA J.B.T. 1 AM 09 - CONTRIBUTION TO HEALTH SURVEILLANCE RELATED TO THE 1. Universidade Federal de Santa Maria, Santa Maria - BIOLOGICAL TOXICITY TEST IN MALE RS, Brazil; 2. Universidade Federal do Pampa, CONDOMS Caçapava do Sul - RS, Brazil; 3. Instituto D’Or de Pesquisa e Ensino, Rio de Janeiro - RJ, Brazil; 4. RIBEIRO, C.¹; CALIL, R¹; ABRANTES, S²; VIDAL, Universidade Federal do Rio de Janeiro, Rio de Janeiro M.¹ - RJ, Brazil.

¹Laboratory of Toxicology, Department of Introduction: Diphenyl diselenide is an Pharmacology/toxicology, National Institute for organoselenium compound which is used as an Quality Control in Health of the Oswaldo Cruz intermediate in organic synthesis. This compound has Foundation (INCQS/FIOCRUZ), Rio de Janeiro, RJ been investigated for its toxicological and ²Laboratory of Chemistry, Department of Chemistry pharmacological properties which is in part associated INCQS/FIOCRUZ, Rio de Janeiro, RJ. with the interaction of diphenyl diselenide with thiols. However, the molecular mechanism(s) involved in INTRODUCTION An area of extreme importance for diphenyl diselenide toxicity are poorly understood. Health Surveillance is the development of research Aim: In the present study, we used a Saccharomyces into products for the protection of the health of the cerevisiae knockout collection to identify gene population, and of these, materials made from natural products that modulate diphenyl diselenide cellular rubber latex, which come into direct contact with the toxicity. Materials and Methods: Haploid S. cerevisiae human body. In this case, because of its importance in single-gene-deletion mutant collection was screened to preventing conception, sexually transmitted diseases identify strains exhibiting altered growth in the and HIV, we can mention the male condoms made of presence of diphenyl diselenide (0.8, 1.6, 3.1, 6.3, 12.5 waterproof natural rubber latex material in order to µM) relative to wild type. The S. cerevisiae single- prevent the passage of body fluids during sexual gene-deletion mutant collection was composed of 167 intercourse In accordance with the standard strains with deletions in genes involved in stress DRC/ANVISA (Resolution of the Collegiate Board of response and metabolism. Results: Among the deletion Directors) no. 62/2008, these products must be tested mutants tested, rad51Δ, rad55Δ, rad50Δ, rad6Δ, tgs1Δ, by suitable institutions employing attesting to the sod1Δ, oca1Δ, urm1Δ and coq3Δ presented quality of the product, thus allowing the issuance of hypersensitive to diphenyl diselenide. The cellular the certificate of quality ISO 9002. OBJECTIVE: processes found to influence sensitivity to diphenyl Toxicity in male condoms using the in vitro diselenide were: oxidation - reduction process (SOD1), cytotoxicity assay - agar diffusion method. cellular response to oxidative stress (OCA1, URM1), METHODS: Thirty-seven (37) samples of male DNA metabolic process (RAD51, RAD55), DNA condoms from different national and international repair (RAD50 and RAD6), and methylation (TGS1 brands were analyzed by the in vitro cytotoxicity assay and COQ3). Moreover, we demonstrate that treatment - agar diffusion method. RESULTS: In total were with diphenyl diselenide increased ROS production, examined thirty-seven (37) samples, we found that 24 analyzed by the oxidation of dichlorofluorescein, in a (64.9%) presented degree of cytotoxicity equal to 2 concentration-dependent manner in S. cerevisiae wild (mild); 12 (32.4%) presented degree of cytotoxicity is type strain. Conclusion: The present study provides equal to 3 (moderate); 1 (2.7%) showed degree of additional insights into diphenyl diselenide toxic cytotoxicity equal to 4 (severe) and none presented mechanisms or detoxification pathways which degree of cytotoxicity 0 (absence). CONCLUSION: collaborate to the overall understanding of the Therefore, the study of possible toxic effects induced molecular targets of this compound and corroborate by these health products should merit the attention of with data in literature which suggest that diphenyl the Sanitary Surveillance in order to minimize and diselenide may induce DNA damage which, at least in prohibit their use to the maximum, if they have part, can be caused by oxidative stress. In addition, the demonstrated cytotoxic effects. results suggest the possible involvement of Financial support/Acknowledgments: FIOCRUZ methyltransferases in the detoxification of diphenyl diselenide. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 23

Financial support/Acknowledgments: FAPERGS, AM 12 - MURINE LOCAL LYMPH NODE CAPES, CNPq. ASSAY (LLNA): A COMPARISON OF ALTERNATIVE AND TRADITIONAL METHODS AM 11 - CITOTOXICITY TEST AS AN ALTERNATIVE TO ASSESS DERMAL SHIMADA, A.L. 1; MORANDI, L. 1; PESTANA, IRRITATION IN SCHOOL MATERIALS C.B.1

SHIMADA, A.L.1; DA SILVA, M.A.A.1; 1 Tecam Tecnologia Ambienta Ltda, São Paulo – SP DOMICIANO, K.S.M.1; MORANDI, L.L.1; PESTANA, C.B.1 The murine local lymph node assay (LLNA) is an alternative method to the guinea-pig sensitization test 1.Tecam Tec. Ambiental that aims to identify substances that have the capacity to cause sensitization based on the measurement of Among the alternative methods recognized in proliferative responses induced in the lymph node after Normative Resolutions 17 and 18 for replacing animal topical exposure. A comparison of the traditional models in the evaluation of products, methods related method (sensitization test in guinea pigs) and to the potential for Care the ones that cause greater alternative method (LLNA) was carried out using the concern due to technical difficulty and high cost. The fungus Isaria fumosorosea, which can infect different standard for the evaluation of skin irritation (OECD orders of insects (weevils, beetles, aphids, whiteflies, 439) is based on reconstructed human epidermis (RhE) etc.) at all stages and is often isolated from the soil. and uses commercially expensive models that are Guidelines used were the OECD Guideline for the difficult to obtain in Brazil. Among the products that testing of chemicals: Skin sensitization, 406 and 442B. should be evaluated for the potential of dermal For the sensitization test in guinea pigs, 30 albino irritation are the school articles. NBR 15236 is the guinea pigs (Cavia porcellus) were divided into two guideline which establishes the safety this articles and groups: experimental and negative control animals. In indicates in vitro assays internationally validated. the induction period, three applications were Considering: the low hazard of these products and the maintained in contact with the skin for 6 hours with the high cost of RhE, our proposal in this study is to use test substance at concentration of 100% on the left the cytotoxicity test to evaluate dermal irritation and flank (experimental) or demineralized water (control corrosion. Studies that correlate cytotoxicity and group). For the LLNA test, 12 mice (Mus musculus), degree of skin irritation suggest the use of monolayer CBA / J strain were divided into three groups: cells as an initial approach to assess the irritation experimental animals and positive and negative potential of human skin. In the present study, 10 controls. The experimental animals received three samples of school materials were tested for topical application of test substance at concentration of cytotoxicity (according to ISO 10993-5) and dermal 100% on the dorsum of the each ear. The protocol was irritation in rabbits (according to OECD 404). In the based on the design proposed by the BrdU and Kit Cell cytotoxicity test, two cell types were used: fibroblasts Proliferation ELISA-Brdu (Roche Applied Science). (V79) and keratinocytes (HaCat). Of the 10 samples On the two methods, the α-hexylcinnamaldehyde tested in rabbits, 9 showed no dermal irritation. The (CAS No. 101-86-0) was used as positive sensitivity only sample that presented dermal irritation in rabbits control. On the traditional guinea pig test, we observed was reversible in 7 days. Of the 10 samples tested for a positive reaction at the 24 hours persistent until 48 cytotoxicity, 8 were approved in the acceptance criteria hours in one animal of experimental group, sufficient (calculation of LD50> 2000 mg / kg) using fibroblasts to consider the test substance as a potential contact (V79) and 7 using keratinocytes (HaCat). The only sentitizer. In addition, on the LLNA test we observed sample that showed dermal irritation in rabbits was that animals exposed to the test substance showed a also toxic with both cells (DL <2000). In only 1 out of stimulation index higher than 1.6 (SI=3.5), 10 samples (10%), fibroblast (V79) cytotoxicity was corroborating the results obtained in the sensitization not predictive of dermal irritation and in 2 out of 10 test in guinea pigs. Based on these results, we conclude samples (20%), cytotoxicity with keratinocytes that the LLNA test proposed as an alternative method (HaCat) was not predictive. Preliminary results have can be used routinely for identify potential sensitizer shown that our proposal is scientifically consistent and substances. is an alternative of lower cost and simplicity for the Financial support/Acknowledgments: Tecam evaluation of school articles. The extension of this test Tecnologia Ambiental Ltda. to a greater number of samples could confirm our preliminary results. Financial support/Acknowledgments: FAPESP

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AM 13 - DISSEMINATING OECD TG 491 AND AM 14 - IN SILICO EVALUATION OF N- OECD TG 492 TESTS IN BRAZIL SUBSTITUTED AND N,N-DISUBSTITUTED IMIDAZOLE DERIVATIVES: AN INSIGHT MARIN, T.M.1; INDOLFO, N.C.1; SILVA, R.1; INTO ADMET PROPERTIES VALADARES, M.2, CRUVINEL, E.3, SANTOS, C.4, ROCHA, V.5, CARDOSO, A.1, PAGANI, E.1 JARDIM, F. 1; RIGOTTO, C. 1; ZIULKOSKI, A. L. 1; MORISSO, F. D. P. 1; SCHREKKER, H. S. 2 1-LNBio/CNPEM, RENAMA, Campinas, Brazil, 2- LFTC, UFG, Goiania, Brazil, 3-PluriCell Biotech, São 1 Universidade Feevale, Novo Hamburgo - RS, Brasil ; Paulo, Brazil, 4-INTERTOX, São Paulo, Brazil, 5- 2 Universidade Federal do Rio Grande do Sul, Porto Natura Inovação e Tecnologia de Produtos, Cajamar, Alegre - RS, Brasil. Brazil. Compounds expressing the imidazole ring in their This project aims at providing proficiency and structure are of great interest. Several studies describe disseminating the ocular toxicity tests OECD TG 491 antifungal, antiviral, antibacterial and antitumor and TG 492 in Brazil. It is sponsored by the Brazilian properties for these molecules. Failures of absorption, Federal Government (CNPq) on behalf of RENAMA, distribution, metabolism, excretion and toxicity the Brazilian Network on Alternative Methods. It (ADMET) parameters are responsible for high rates of involves five institutions: one private non-profit molecules that do not reach the market. In silico research center (LNBio); one academic research center methodologies have been used as an alternative (Federal University of Goiás); one contract research method to the use of animals with the objective of organization (Intertox); one cell line manufacturer predicting properties of chemical compounds, directly (PluriCell Technologies) and one cosmetic impacting the cost and time of the drug discovery and manufacturer (Natura). These institutions with development process. In this context, the present study different backgrounds and located more than 1,000 Km aimed to explore in silico techniques to predict the apart are connected by an Enterprise Laboratory molecular properties of several N-mono and N,N- Platform – ELP composed of a Laboratory Information disubstituted imidazole derivatives containing cationic Management Systems - LIMS and Electronic nucleus (C1 – C10) in order to identify possible Laboratory Notebook – ELN. This allows remote and molecules with therapeutic potential through ADMET secure access to project data, responsibility assignment calculations. Initially, the molecular structures were and activities monitoring by all organizations involved. elaborated in GaussView 5.0 and submitted to the This consortium’s model provides strong integration pharmacokinetic analysis in the DataWarrior software and delivers better results and might be replicated by and admetSAR web server. A compilation of results others nations. References: Test No. 491: Short Time referring to ADMET descriptors ranked all compounds Exposure In Vitro Test Method for Identifying i) with penetration into the blood-brain barrier. In Chemicals Inducing Serious Eye Damage and ii) relation to the human intestinal absorption and Caco-2 Chemicals Not Requiring Classification for Eye permeability, C7 and C10 did not obtain positive Irritation or Serious Eye Damage.Test No. 492: absorption, respectively. C3, C7 and C9 showed high Reconstructed human Cornea-like Epithelium (RhCE) inhibitory promiscuity of cytochrome P450 enzymes, test method for identifying chemicals not requiring increasing the chance of toxicity by pharmacokinetic classification and labelling for eye irritation or serious interactions to drugs that are metabolized by these eye damage. enzymes. Of the compounds studied, C1-C9 have been Financial support/Acknowledgments: CNPq. classified as an inhibitor of the transport system of organic cations and could thus interfere with excretion phase of xenobiotics. For the toxicological profile, only C1 and C2 presented toxicity by the AMES test. None of the derivatives were considered carcinogenic and only C10 showed low irritant capacity. All compounds were classified in category III of acute oral toxicity based on US EPA criteria, suggesting slightly toxic compounds. Results indicated that C4, C5, C6 and C8 imidazole derivatives showed a satisfactory predictive ADMET profile, subject to structural optimization to become potential drug candidates. Financial support/Acknowledgments: Feevale University.

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AM 15 - EVALUATION OF THE ARTEMIA AM 16 - APPLICABILITY OF MONO MAC-6 SALINA (BRANCHIPUS STAGNALIS) CELL LINE IN THE MONOCYTE ACTIVATION BIOLOGICAL RESPONSE AGAINST THE TEST (MAT) AS AN ALTERNATIVE METHOD FENTON REACTION AND BHT TO THE RABBIT PYROGEN TEST (RPT) IN THE ROUTINE OF THE QUALITY CONTROL ARAUJO, V.A.P.1; DIAS, A.J.S.2; NUNES, A.R.2; LABORATORY BATISTA, E.F.2; SANTOS, D.C.2; SILVA, G.A.1,2 GIMENES, I. 1; VIEIRA, D. H. 1; OLIVEIRA, 1. Universidade Federal do Amapá, Programa de Pós- C.B.N. 1; CALDEIRA, C. 1; PRESGRAVE, O. 1. graduação em Ciências Farmacêuticas PpgCF, UNIFAP, Macapá, Amapá; 2. Universidade do Estado 1. Instituto Nacional de Controle de Qualidade em do Amapá, Laboratório de Química Orgânica e Saúde/FIOCRUZ, Rio de Janeiro - RJ, Brasil. Bioquímica, UEAP, Macapá, Amapá. Introduction: Pyrogens can induce inflammatory Oxidative stress is the imbalance between antioxidants reactions, including fever and more severe adverse and reactive oxygen species (ROS) in which they reactions, such as septic shock and death. This kind of cause cell damage, among these damages the contamination is considered a great public health lipoperoxidation (LP) process due to the main action of problem. Pyrogen tests are safety assays performed in the hydroxyl ions. Thus, the objective of this study was the routine of quality control of injectable products by to evaluate the response of Artemia salina (Branchipus regulatory agencies. The Rabbit Pyrogen Test (RPT) is stagnalis), a microcrustacean that was used as test based on the fever reaction of rabbits, since rabbit’s organism (TO), in front of the Fenton reaction (FR), response is the same as that found in the human fever. capable of generating ROS. The test method that Since 2010, five in vitro tests called as Monocyte quantifies the oxidative damage in cellular lipids by Activation Test (MAT) are included in the European levels of thiobarbituric acid reactive substances Pharmacopoeia as a third method for detection of (TBARS) has been used, thus proposing an alternative pyrogen. The MAT is an in vitro test based on the method to the use of chordate animals for evaluation of activation of human monocytes by pyrogens and antioxidant compounds. For that, three groups were quantification of cytokine release. The MAT can be made: control; TO + FR; TO + FR + standard performed with different source of monocyte cells. antioxidant. The incubation was done according to However, the use of Mono Mac 6 cell line (MM6) has Persoone et al (1989), 0.300 g of TO were added in the advantage of not depending on volunteers. 400 ml of artificial salt water, leaving them for 24 h in Objective: The aim of this study was to evaluate the the water bath (30 ± 1 ° C), with artificial lighting and applicability of MM6 cell line as an alternative method aeration constant. For the induction of peroxidation by to detect pyrogens through comparative analysis with Fenton reaction, 5 mL of the FeSO4 solution was in vivo and in vitro model. Material and methods: transferred to the volumetric flask and another 351.87 Negative control and Lipopolysaccharide (LPS) from μL of H2O2 (10%) was added with brine to the 100 Escherichia coli O55:B5 in concentrations between mL measure. In order to measure the concentration of 0.25 and 20.0 EU/mL were used to the dose-response the final product of LP, the methodology described by and concentration-response curve. Rabbits were VanYe et al (1993) was modified. BHT was used as injected with LPS doses and the rectal temperature the standard antioxidant. As results, TBARS levels in were measured during 3 hours. For MAT a pool of TO incubated with standard antioxidant had the lowest human whole blood cryopreserved (Cryo-WB) and value (3.08±0.15mmol/mL), when compared to levels MM6 cells were incubated with LPS concentrations in TO incubated with the Fenton reaction (8.61 ± 0.16 and IL-1β and IL-6 (EUmL-1) releases were measured mmol /mL), which showed a high level of by ELISA. Results: The parallel testing analysis lipoperoxidation and the TBARS levels of the control showed a relationship between the results obtained in group (6.27 ± 0.18 mmol / mL) that underwent vivo and in vitro tests. The RPT demonstrated a well- autoxidation of the cellular components. Alternative defined dose response curve with a temperature methods to animal use that cite the use of Artemia increase of ≥0.5 °C. A positive correlation in the salina for evaluation and analysis of the antioxidant release of cytokines in the presence of increasing doses activity of natural products have not yet been reported of LPS were observed both human whole blood in the literature. The advantage of this alternative cryopreserved and MM6 cells. Conclusions: The MAT method over the already described methods lies in the using MM6 cells is a rapid and reliable alternative biological medium of easy manipulation and low method compared to RPT. Thus, it could be used as an economic cost. alternative method to replacement of a RPT by the Brazilian regulatory agencies. Financial support / Acknowledgments: INCQS/FIOCRUZ ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 26

AM 17 - THE PRE-CLINICAL ASSESSMENT OF AM 18 - A NOVEL TWO-STEP HIERARCHICAL A TOBACCO HEATING PRODUCT (GLOTM) QUANTITATIVE STRUCTURE-ACTIVITY RELATIONSHIP FOR PREDICTING HUMAN MURPHY, J. 1; THORNE, D. 1; BREHENY, D. 1; SKIN SENSITIZATION GACA, M. 1; LIU, C. 1; PROCTOR, C. 1. BORBA, J.V.V.B. 1; ALVES, V. M. 1. 2.; SILVA, A. 1. British American Tobacco, R&D Centre, C. 1.; MURATOV, E. 1. 2.; LUETCHTEFELD, T. 3.; Southampton SO15 8TL, United Kingdom HARTUNG, T. 3.; TROPSHA, A. 2.; ANDRADE, C. H. 1. Introduction: Tobacco heating products (THPs) represent a subset of next-generation nicotine and 1. Laboratory for Molecular Modeling and Drug tobacco product category, in which tobacco is heated Design, Faculty of Pharmacy, Federal University of at temperatures of 250-350°C instead of burning Goias, Goiania, GO, Brazil; 2. Laboratory for (900°C), having the potential to significantly reduce Molecular Modeling, Eshelman School of Pharmacy, cigarette smoke toxicants. THPs holds great potential University of North Carolina, Chapel Hill, NC, USA; for reducing the harm associated with tobacco use but 3. Center for Alternatives to Animal Testing (CAAT), needs to be scientifically proven. Objective: To Johns Hopkins Bloomberg School of Public Health, characterise the aerosol emissions and assess the Baltimore, MD, USA. biological impact of the novel THP (gloTM) comparing results to a reference 3R4F cigarette. Introduction: Skin sensitization is a common adverse Materials & Methods: Using a newly-developed 5-step reaction caused by repetitive exposure to a chemical assessment, THP product designs were evaluated to allergen. Recently, studies have shown that the eliminate combustion and generate an aerosol by preferred animal test for skin sensitization, LLNA, heating tobacco. This was confirmed by using targeted does not correlate well with human potency in some chemical analysis across several public health toxicant cases. However, despite the ban on animal testing for lists including the WHO’s TobReg9 priority toxicants. cosmetics, publicly available animal data is still A suite of in vitro toxicological assays were used to growing and there is a strong need to improve the assess the mutagenic potential of particulate extracts usefulness of this data. Objectives: We aimed to (TPM) and aerosols (WA) including the Ames assay develop and rigorously validate a two-step hierarchical using 5 Salmonella typhimurium tester strains and the Quantitative Structure-Activity Relationships (QSAR) mouse lymphoma assay. The Bhas 42 cell approach to predict human skin sensitization. Materials transformation assay was incorporated for detecting and methods: We have (i) compiled, curated, and tumour promoters and the neutral red uptake cell integrated four datasets comprising 1,027 chemicals viability assay provided an acute measure of tested in the LLNA and 138 compounds with human cytotoxicity. Contemporary screening approaches data; (ii) partitioned LLNA data into those for which employing a luciferase-based reporter gene assay and the response agrees with human response and those multiparametric analysis incorporating 10 different that does not; (iii) built binary QSAR models that toxicity and oxidative stress endpoints were used to could discriminate between compounds in these two assess total particulate matter (TPM) from products. classes; (iv) finally, we have established rigorous and Results: The results show that this product eliminates externally predictive QSAR models of LLNA for the tobacco combustion. Targeted chemical analysis dataset comprehending 1,027 chemicals. Results and demonstrates over 90% reduction in aerosol toxicant discussion: The models generated using Morgan emission compared to cigarette smoke. Compared to descriptors and Random Forest algorithm were cigarette smoke, THP demonstrates significantly predictive with validation metrics above threshold of reduced toxicological and biological responses across 0.6. The developed two-step hierarchical QSAR all endpoints tested. Conclusion: The data generated modeling workflow improved prediction of human add to growing evidence that suggests that THPs may response using only LLNA data by almost 20%. provide a safer alternative to traditional cigarettes, Conclusions: We demonstrated the power of using however further studies investigating the longer terms integrative QSAR modeling for identifying chemicals effects on consumers is required to substantiate disease that are either safe or toxic in humans, thus providing relevant risk reduction. strong support for exploring reliable alternatives to animal testing for human skin sensitization. Financial support / Acknowledgments: CNPq e CAPES.

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 27

AM 19 - IN VITRO STUDY OF Financial support / Acknowledgments: FAPDF, ANTIMICROBIAL AND HOST-DEFENSE CNPq, CAPES. PEPTIDES FOR HUMAN TISSUE REGENERATION AM 20 - CYTOTOXIC ACTIVITY OF RHEIN SILVA, T. A.1; CARDOSO, A. G.1;SILVA, L. P. S.1; THE MAJOR HUMAN METABOLITE OF BRAGA, M. C.1; SANTANA, G. O. S.1; ANDRADE, DIACEREIN R. V.; DIAS, S. C.1; FRANCO, O. L.12; CARVALHO, J. L.1 JAPIASSU, K. B.1; FERREIRA, J. M. U. 1; ARRUDA, E. L. 1; CARVALHO, P. H. O.1; 1. Universidade Católica de Brasília, Brasília - DF, CORTEZ, A. P. 2; VALADARES, M. C. 2; LIÃO, L. Brasil; 2. Universidade Católica Dom Bosco, Campo M. 3; OLIVEIRA, V. 1 Grande - MS, Brasil. 1 Laboratório de Bioconversão– Faculdade de Introduction: A requirement for safety assessment of Farmácia – Universidade Federal de Goiás; 2 any potential cosmetic product applied to the skin is Laboratório de Farmacologia e Toxicologia Celular – essential. Decreasing acceptance of animal use in Faculdade de Farmácia – Universidade Federal de cosmetic testing paves the way for human tissue Goiás; 3 Laboratório de Ressonância Magnética cultures as an alternative. In this sense, the present Nuclear – Instituto de Química – Universidade Federal work performs, for the first time, the evaluation of de Goiás. cytocompatibility and regeneration potential of synoeca-MP and IDR-1018 in human material. Such Introduction: Diacerein (1,8-diacetoxy-3- peptides have shown antimicrobial and host defense carboxyanthraquinone) is an anthraquinone with a properties in animal models. Methods: To validate wide range of biological activities, like as anti- peptide’s antimicrobial potential, the minimum osteoarthritis, analgesic, anti-inflammatory, inhibitory concentration (MIC) against Staphylococcus antipyretic, prevention of vascular disease, insulin aureus was determined. Peptide cytotoxicity on human resistance treatment, anticancer. Biotransformation fibroblasts (hFib) was evaluated using MTT assay. studies are important in the evaluation of safety and Regenerative potential was assessed by migratory efficacy and for developing of new drug candidates. behavior evaluation of the fibroblasts using the scratch The "microbial models of mammalian metabolism", in method and collagen contraction assay, performed by which microbial biotransformation is used for the measuring the area of COL-I gels containing hFibs. purpose of predict and obtaining human metabolites, is Results: Current results indicate that IDR-1018 has no an alternative method to the use of animals for this bactericidal activity on S. aureus, unlike synoeca-MP, study. Several advantages such as lower cost, a greater which was able to inactivate these bacteria (>16 quantity and variety of derivatives produced, using µg.mL-1). However, at this concentration, synoeca-MP mild conditions of reaction and decreasing the use of slightly decreased the hFibs viability (81.0% ± 3; toxic volatile organic solvents has been observed. Aim: p<0.05), while IDR-1018 did not significantly The aim of this study was to produce in vitro the compromise cell viability (94.3% ± 1.62). When human metabolite of diacerein by biosynthesis combined, IDR-1018 reduced the cytotoxic effect of catalyzed by filamentous fungi and to evaluate the synoeca-MP (85.8% ± 6.42). It was observed that IDR- cytotoxicity of rhein. Methods: Screening with 1018 increased hFibs migratory behavior compared to seventeen fungal strains was realized in microscale SBF, which was used as positive control (17.3% with PDSM medium for 168 hours at 30° C and 150 control x 27.1% IDR-1018). The COL-I contraction rpm. The characterization was done using techniques behavior was observed and the gels containing the as Nuclear Magnetic Resonance (NMR1H13C), High associated peptides showed greater contraction Resolution Mass Spectrometry (MS), spectrometry in compared to other groups. Real-time PCR analysis of the UV region and analysed by HPLC. It was added an the cells showed that Ki67 is induced both by IDR- inhibitor of CYP450 after 65 hours of growth. The 1018 and IDR-1018 combined with Synoeca, while cytotoxic potential of diacerein and rhein was BCL2 is increased after IDR-1018 treatment. CXCR4 evaluated by the tetrazolium reduction method (MTT) and CXCR7 genes had their expression increased assay using murine fibroblast cells 3T3 and tumor cell when hFibs were treated with IDR-1018. Conclusion: line B16F10. Results: After screening, Aspergillus The results obtained so far indicate the potential of in ochraceus ATCC 1009 was selected for incubation in vitro human cell cultures as an alternative to the semipreparative scale and was obtained the major animal tests. In addition, when combined, synoeca-MP human metabolite characterized as rhein. The and IDR-1018 have shown great potential as an influence of cytochrome P450 inhibitor in the antimicrobial and pro-regenerative treatment for skin production of rhein showed the reduction of 41%, lesions. which may indicate the involvement of CYP1A1 and ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 28

CYP1A2 in the deacetylation reaction. Rhein presence of N-oxide metabolite, with the loss of demonstrated cytotoxic potential for melanoma cells piperazine moiety after fragmentation. Desmethyl- (B16F10) with IC50 67μM, within 24 hours, and IC50 olanzapine was equally identified and characterized. 58 μM, within 48 hours. Conclusion: A. ochraceus was The yield of the reaction was very efficient 20.0% able to produce rhein, major human metabolite of when compared with microsome methods. These diacerein, which showed to have cytotoxic activity in results demonstrate the interesting potential of such B16F10 cells. alternative methods for preparation of functionalized Financial support / Acknowledgments: CAPES. derivatives compared to other methods. Financial support / Acknowledgments: CAPES.

AM 21 - IN VITRO ALTERNATIVE METHOD FOR OLANZAPINE REFERENCE STANDARDS AM 22 - IN VITRO DIFFERENTIATION OF PRODUCTION EMBRYONIC STEM CELL FROM MURINE DURING EMBRYOID BODIES FORMATION JAPIASSU, K. B.1; SOUZA, P. L. M. 1; ARRUDA, E. L. 1; BASTOS, C. C. C. 1; LEONEL, F. O. 1; VAZ, FARIA, L.C.1; SANTANA, D.P.1; GUILLO, L.A. 1; B. G. 2; OLIVEIRA, V. 1 1. Universidade Federal de Goiás, Goiânia - GO, 1 Faculdade de Farmácia - Universidade Federal de Brasil. Goiás, Goiânia – GO, Brasil; 2 Instituto de Química - Universidade Federal de Goiás, Goiânia – GO, Brasil. Embryonic stem cells are derived from the inner cell mass of the blastocyst and when cultured under Olanzapine belonging to the class of appropriate conditions, can differentiate into all three thienobenzodiazepine, is the newer atypical germ layers: ectoderm, mesoderm and endoderm. antipsychotic drugs, indicated for treating of Embryoid bodies are three-dimensional aggregates of schizophrenia, bipolar disorder and related psychoses. embryonic stem cells and exhibit several properties Despite its effect in the treatment of these diseases and distinct from monolayer cells, which resemble the the increasing prescription, over 80% of patients do early stages of embryonic development. Gene not continue the treatment due to its adverse events. expression during the formation of embryoid bodies Filamentous fungi are able to metabolize drugs in has been investigated in several studies. Thus, this human metabolites across its enzymatic arsenal similar study evaluated the expression of genes related to the to the mammalian, in mild reaction conditions and undifferentiated state (Oct4 and Nanog), genes related without requiring costly culture medium. to the differentiation in ectoderm, endoderm and Determination and quantification of these drugs and mesoderm (Nestin, Alpha-feto protein, Bmp4, their metabolites present many applications clinical, respectively) and Gapdh (constitutive gene, positive toxicological and forensic, with wide demand of control of the PCR reaction) during differentiation of reference standards. The aim of this study was the embryonic stem cells in the three-dimensional culture production in vitro of olanzapine metabolites to model. Murine embryonic stem cells were maintained evaluate pharmacokinetic processes. In order to obtain in their undifferentiated state and then transferred to olanzapine metabolites, spore suspensions of the AggreWell TM400 plate, where they were Beauveria bassiana ATCC 7159 were sown in liquid incubated for 72 hours in order to form the embryoid culture medium PDSM, after 65 hours, at 27 °C and bodies. Subsequently, the formed embryoid bodies 200 rpm, a solution with 50 mg∙mL-1 of Olanzapine were incubated for 12 days in 24-well plate to continue was added to the reaction medium and the Erlnemeyers the differentiation process. Gene expression analysis were maintained in the same conditions. Aliquots of was done by RT-PCR and visualization of amplicons the reaction medium were taken every 24 hours and was performed by agarose gel electrophoresis. Our submited to TLC analysis. At the end of reaction, results showed that after 24 hours of incubation on the medium was saturated with NaCl and filtered through AggreWell TM400 plate, the cells formed aggregate Celite®, proceeding by the extraction with ethyl structures. At the end of 72 hours, the embryoid bodies acetate, methanol and acetone. The extracts were formed were visually uniform in size and shape. After concentrated in vacuum and preceded to analysis by 12 days of incubation, the embryoid bodies began the Fourier-transform Ion Cyclotron Resonance Mass process of differentiation for ectoderm and mesoderm Spectrometry High Resolution (FT-ICR MS). The in linages, however still showed low levels of expression vitro production of olanzapine human phase I of Nanog gene, indicating that the process of embryoid metabolites by B. bassiana was achieved in free- bodies differentiation was not complete in this period. whole-cell biosynthesis. Their characteristic fragments Alpha-feto protein gene was not expressed, showing in FT-ICR MS analysis observed Desmethyl and N- that the 12 days period was not sufficient to induce the oxide; m/z 329.13635→m/z 229.06078 indicates the differentiation of embryoid bodies to endoderm ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 29 lineage. Through the in vitro test of formation of Financial support / Acknowledgments: FIOCRUZ embryoid bodies and the use of molecular markers, it was possible to observe the expression of genes involved in the process of cell differentiation of AM 24 - IN VITRO ALTERNATIVE METHOD embryonic stem cells. FOR PRODUCTION OF STANDARD Financial support / Acknowledgments: CNPq, PROPOFOL METABOLITE PHASE II AND ITS PRONEX/FAPEG and CAPES/PNPD. APPLICATION IN PHARMACOKINETIC STUDIES

CARVALHO, P.H.O.1; NÓBREGA, G. C. 1; AM 23 - CYTOTOXICITY TESTS FOR ARRUDA, E. L. 1; JAPIASUL, K. B. 1; OLIVEIRA, EVALUATING MEDICAL DEVICES: AN V. 1 ALERT FOR DEVELOPING BIOTECHNOLOGY HEALTH PRODUCTS 1 Universidade Federal de Goiás, Goiânia - GO, Brasil. VIDAL, M.N.P.1, GRANJEIRO, J.M.1 Introduction: Propofol (2,6-diisopropylphenol) is an 1.FIOCRUZ intravenous anesthetic used worldwide for the induction and maintenance of anesthesia in many contain a certain degree of risk, which requires strict animal species. It is also used for prolonged sedation in quality control in their production, distribution and use intensive care units, mainly due to favorable as well as in the disposal of their waste in the pharmacological properties, short onset and rapidly environment. With continuous development in science reversible hypnosis. The biotransformation of propofol and technology, medical devices must undergo occurs in the liver via hydroxylation by the enzymatic intradermal irritation and testing for sensitization, CYP450 complex and via glucuronidation by UGT cytotoxicity, and acute systemic toxicity. In health (UDP-glucuronosiltransferase) as phase II metabolism. care, biotechnology aims to provide technology based Filamentous fungi can be used for biotransformation products or processes related to energy, food and studies as an alternative way to identify and produce health, which are capable of stimulating new mammalian metabolites, mimicking in vivo hepatic businesses, expanding exports, integrating into the reactions of the drugs. These microorganisms can value chain, and stimulating new demands for contribute to the elucidation of metabolic routes of the innovative products and processes. Objective Our compounds, providing information on the generation objective was to study which in vitro cytotoxicity of more active, inactive, toxic or less toxic substances. assays are commonly used to evaluate medical devices, Aim: The aim of this study was to produce in vitro the as an alert for the greater reliability of these health phase II metabolite of Propofol using microorganisms. products. Methodology The literature review was Methods: In order to evaluate the biotransformation conducted in November 2016 through an online search profile of Propofol, the fungus Cunninghamella of the following scientific electronic databases. The elegans ATCC 26169 was selected. Biotransformation keyword searched was biomaterials, and because of the was performed in the PDSM culture medium, 200rpm, thousands of papers found, the search was refined 27ºC. The monitoring of the reaction kinetics was done using the following keywords, which were spelled by High Performance Liquid Chromatography. The either in English or in Portuguese: derivate was elucidated by High Resolution Mass biomaterial/biomaterials, Spectrometry (ESI-MS). Results: The spectra obtained biocompatibilidade/biocompatibility and by ESI-MS showed a molecular ion [M + Na]+ m/z citotoxicidade in vitro/cytotoxicity in vitro. Results 363,1781 characterizing the presence of a hydroxyl The online search performed in all three databases and a glucose in the molecule of propofol showing the resulted in finding 3,366 papers, of which 1,006 (30%) formation of compound 1-hydroxypropofol- were related to the descriptor Biomaterial, while 1,765 glucosideo. Conclusion: It is possible to produce in (52%) were related to Biomaterials × Biocompatibility, vitro a Phase II metabolite of Propofol using 439 (13%) to Biomaterials × Cytotoxicity in vitro, and filamentous fungi. This suggests a presence of the 156 papers (5%) to Biomaterials × Biocompatibility × enzyme glycosyltransferase in the microorganisms in Cytotoxicity in vitro. CONCLUSIONS Cytotoxicity question. A simultaneous pharmacokinetic study of tests are one of the most important indicators of an in Propofol in mammals indicated the presence of the vitro biological evaluation system. Tests on cell same metabolite in animal samples, confirming the cultures can be successfully performed because they importance of applying this model to obtain and are fast, reproducible, sensitive and affordable for an in predict mammalian metabolites as an alternative vitro biocompatibility study. Moreover, they represent method for animal experimentation. This product an important step in the development of new products obtained served as a standard for an evaluation of by evaluating cytotoxic effects. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 30 pharmacokinetic profile of Propofol during anesthesia Financial support / Acknowledgments: Fundação of sheep. Oswaldo Cruz (FIOCRUZ) Financial support / Acknowledgments: CAPES.

AM 26 - THE IN VITRO PHOTOPROTECTIVE AM 25 - IN VITRO CYTOTOXICITY TEST OF POTENTIAL OF CAROTENOIDS AND OTHER AGAR DIFFUSION IN THE QUALITY PIGMENTS FROM ANTARCTIC BACTERIA CONTROL OF SURGICAL GLOVES AND PROCEDURE TAVARES, R.S.N.1; SILVA, T. R. 3; COLEPICOLO, P. 2; DEBONSI, H. M.1; DE OLIVEIRA, V. M.3; CALIL LEMOS, R.1; VIDAL, M.N.P.1; ZAMITH, GASPAR, L. R.1 H.P.S.1 1 Universidade de São Paulo, Faculdade de Ciências 1. Laboratory of Toxicology, Department of Farmacêuticas de Ribeirão Preto, Ribeirão Preto - SP, Pharmacology and Toxicology, INCQS / FIOCRUZ, Brasil; 2. Universidade de São Paulo, Instituto de Rio de Janeiro, RJ, Brazil. Química, São Paulo - SP, Brasil; 3. Universidade Estadual de Campinas, Campinas -SP, Brasil. INTRODUCTION: The occurrence of irritative type contact dermatitis in healthcare professionals using Antarctic continent is considered a unique biome, surgical gloves and procedure indicates the importance which means big part of its biological patrimony of including in the Brazilian legislation cytotoxicity cannot be found elsewhere on Earth. In such tests to evaluate this type of product as one of the conditions, is expected to find extremophiles requirements to ensure its safe use. With the microorganisms to be investigated due its unique increasingly strict control over the use of laboratory metabolites with diverse biotechnological applications. animals, the use of in vitro tests with mammalian cells In this context, presenting great market importance, we is necessary to detect the toxicity of medical and can mention carotenoids. Several of them with hospital devices in direct contact with human tissue. photoprotective activities were isolated from Antarctic Among these tests, the in vitro cytotoxicity test of agar bacteria in the last century (1). Thus, the aim of this diffusion in NCTC mouse fibroblast cells clone L-929 study was to evaluate the photoprotective potential of can be used as a first step in the biological evaluation pigments isolated from different species of Antarctic of the glove material. Objective: To evaluate the bacteria. However, during the natural products toxicity of surgical latex gloves and procedure gloves screening for new candidates to sun protection, the (vinyl and nitrile) using the in vitro cytotoxicity assay - photodegradation (UVA irradiation dose of 27,5 agar diffusion method. MATERIALS AND J/cm2) and phototoxicity (measured in 3T3 fibroblast METHODS: The thirty-five glove samples used in 2 cultures for the determination of cell viability in the sentinel hospitals and at a FIOCRUZ research unit, presence and absence of UVA radiation (2) are negative control (United States Pharmacopeia-USP requested. At this point, a major indication of the non- reference material), and positive control (latex garrote) phototoxicity of a substance is the maintenance of the were tested on cultures of L-929 In duplicate. Samples molecular structure after solar irradiation, i.e. of gloves and controls (about 100 mm2) were placed photostability. Eight extracts of different species of on the solidified surface of 0.9% agar with 0.05% Antarctic pigmented bacteria were analyzed: 50cyt, neutral red on the cell monolayer 48h after 235, 416, 465, 466, 366, 424, 395. Four of them establishment of the cultures. The degree of presented UVA/VIS absorption (50cyt, 416, 465 e cytotoxicity (0 to 4) was evaluated 24 hours after 366) and one of them absorbed on UVB range as well application of the samples. RESULTS: In the tests, the (50cyt). All of the four studied extracts were negative control showed absence of cytotoxicity considered photounstable in the UVA/VIS range; (degree 0) and the positive control, moderate however, the 465 and 366 pigments remained stable in cytotoxicity (grade 3). All 28 latex gloves had a degree the UVB, since there was a slight reduction in the of cytotoxicity 3 which would disapprove the samples UVB range. The four pigments tested were considered according to USP 39th Edition; the 4 nitrile gloves and phototoxic. Regarding three analyzed 50cyt fractions, the 3 vinyl ones would be approved in the test because obtained by HPLC (that was the most intense they presented mild cytotoxicity (grade 2). No glove absorbance signal); 50cyt18 was considered cytotoxic showed degree 0 of cytotoxicity. CONCLUSION: The (IC: 26.5 µg/ mL), 50cyt23 was considered phototoxic test was able to discriminate the toxicity of latex (Mean Photo Effect - MPE: 0.931/0,909) and 50cyt28 gloves compared to nitrile and vinyl gloves which was considered non-phototoxic (MPE: 0,088/ -0,034), were considered safer for human use and with less demonstrating how the separation strongly contributes potential to cause adverse reactions. to obtain different results. Following studies with their isolated and identified compounds should be ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 31 conducted to evaluate which molecules are responsible Moreover, the toxicophorical screening revealed low for the UV absorption and if the toxicity potential potential hazards, revealing that ellagic acid is an would be further reduced with purification. interesting natural product starting point for obtaining Financial support / Acknowledgments: CNPq, novel proton pump inhibitors for gastric ulcer FAPESP. treatment. Financial support / Acknowledgments: FAPEG e CAPES. AM 27 - EVALUATION OF MECHANISM OF ANTIULCER ACTIVITY AND TOXICITY OF ELLAGIC ACID BY IN SILICO APPROACH AM 28 - NON-RELEVANCE OF ACUTE DERMAL TOXICITY TESTING FOR SOUSA, S. A. 1; OLIVEIRA, M. G. 1; VAZ, P. D. S. ASSESSING HUMAN HEALTH PROTECTION 1; PAULA, J. R. 1; SILVA, V. B. 2; BARA, M. T. F. 1 IN THE REGULATORY DECISION MAKING FOR AGROCHEMICAL FORMULATIONS 1.Laboratório de Pesquisa de Produtos Naturais-LPPN, Faculdade de Farmácia, Universidade Federal de LATORRE, A. O. 1; SOUZA, J. C. P. 2; BOFF, M. M. Goiás, Goiânia - GO, Brasil 2. Escola de Ciências 3; FAGUNDES, P. M. 4; MARTINS, A. P. 5; Médicas, Farmacêuticas e Biomédicas, Pontifícia CATALANO, S. M. I. 6; CAZARIN, K. C. C. 6; Universidade Católica de Goiás, Goiânia - GO, Brasil. ROMEO, L. L. 1

Introduction: Peptic and/or duodenal ulcers are 1 Bayer S.A., Sao Paulo, SP, Brasil; 2 Dow characterized by diverse acute and chronic ulcerative AgroSciences Industrial LTDA, Sao Paulo, SP, Brasil; lesions that commonly arise in any portion of the 3 IHARABRAS S.A. Indústrias Químicas, Sorocaba, gastric mucosa that is exposed to the aggressive action SP, Brasil; ; 4 Syngenta Crop Protection, Sao Paulo, of gastric acid. The modern approach to control SP, Brasil; 5 DuPont do Brasil S.A., Paulínia, SP, gastroduodenal ulceration is to inhibiting H+-K+- Brasil; 6 BASF S.A., Sao Paulo, SP, Brasil. ATPase pump to control increase acid secretion. The Brazilian species Lafoensia pacari have popular and A recent EPA publication (2016)1 showed that dermal pharmacological reports on the treatment of stomach toxicity tests compared to oral toxicity tests offer little disorders. Previous studies have revealed the presence or no additional scientific information or public health of elagitannins in this species being ellagic acid, one of protection and describes guidance for waiving acute the most important pharmacological active dermal LD50 studies for agrochemical formulations. It compounds. Objective: To evaluate the mechanism of also points out the importance of avoiding unnecessary antiulcerogenic activity of ellagic acid and to use of time and resources, data generation costs and investigate its toxicological potential by in silico tools. animal testing. Based on EPA publication, the Materials and Methods: Docking simulations with Technical Group on Toxicological Risk Assessment ellagic acid and gastric proton pump (PDB: 5Y0B) (GT-ART) has consolidated data from some were run with Dockthor server. The calculations were companies associated to the Brazilian Crop Protection performed inside a sphere of 7 Å radius centered at the Association (ANDEF) to obtain information on results O atom of the Ala335 residue. Toxicity prediction of acute oral and dermal LD50 studies of agrochemical procedure with ellagic acid was performed with formulations registered in Brazil. A retrospective ACD/I-Lab, Lazar, Pred-hERG and Pred-Skin online analysis of this broadly representative dataset was tools. Results: Docking simulations within proton carried out by GT-ART, following these pump revealed that ellagic acid was capable of considerations: only rat studies were selected; only accommodating in the active site of protein by paired studies were chosen; only studies performed interacting primarily with Asn138 through hydrogen with top doses 2000 mg/kg were selected; bond and Tyr799 with pi-shaped and Ile816 with pi- formulations of Biological products were excluded. alkyl interactions, which are considered some key The dataset includes 351 formulations in 21 residues for the proton pump inhibition. Prediction formulated product types. Among these 351 data revealed no toxicological potential for most of the formulations, 237 formulations have a single active parameters evaluated with non-detected risk regarding ingredient, 107 have two and 7 have three or more. AMES toxicity, carcinogenicity, hERG inhibition and The comparison of toxicity between oral and dermal binding to the estrogen receptor. On the other hand, studies revealed 188 of the 351 (53.6%) formulations only a predicted risk of skin sensitization was detected. with the same toxicity category in both studies, 136 Conclusion: In silico findings show the hypothesis of formulations (38.7%) have oral studies with more ellagic acid presenting antiulcer activity by interacting restrictive category than dermal studies and 27 in the H+-K+-ATPase pump active site, thus formulations (7.7%) have dermal studies with more contributing to the reduction of acid production. restrictive category than oral studies. For these 27 ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 32 formulations, 23 formulations have a toxicity Category AM 30 - PHARMACOKINETICS OF 5 for dermal and were Not classified for oral, likely as ACETAMINOPHEN IN A 2-ORGAN-CHIP (2- a consequence of dose selection of 4000 mg/kg and not OC) PLATFORM due to toxicity from dermal exposure below 5000 mg/kg. Resulting in 4 formulations (1.1%), which MARIN, T.M.1; INDOLFO, N.N.1; ROCCO, S.A.2; dermal studies determined more restrictive category CARDOSO, A.C.1, PAGANI, E.1 than oral studies. The results presented here corroborates EPA conclusion on waiving dermal 1-LNBio/CNPEM, RENAMA, Campinas, Brazil, , 2- toxicity tests for agrochemical formulations. 1US EPA. LNBio/CNPEM ,NMR Laboratory, Campinas, Brazil. (2016) Guidance for Waiving Acute Dermal Toxicity Tests for Pesticide Formulations & Supporting Microfluidic systems are promising for developing Retrospective Analysis. better non-animal tests, but pharmacokinetic (PK) data Financial support / Acknowledgments: Associação in chips are missing. Acetaminophen (APAP) is 95% de Defesa Vegetal (ANDEF). conjugated by the liver before renal excretion, making the liver the main organ for APAP metabolism. This study addressed the APAP PK properties in the 2-OC AM 29 - LONG-TERM SURVIVAL OF (by TissUse GmbH). A 2-OC was assembled with an SPONTANEOUSLY BEATING intestinal barrier of CaCo2 + HT29 cells in one CARDIOMYOCYTE SPHEROIDS A 2-ORGAN- compartment with microfluidic communication with CHIP (2-OC) PLATFORM the other having liver spheroids of HepaRG + Stec Cell. The administration route was emulated by APAP MARIN, T.M.1; CARVALHO, M.1; BIAGI, D. 2; placement on the intestinal barrier (oral) or in the VALADARES, M.2; CONSONI, S.R.3, INDOLFO, media (intravenous). Media samples were collected N.1; PAGANI, E.1 and analyzed by HPLC/UV. Preliminary data show slower absorption through the CaCo2 + HT29 cells 1-LNBio/CNPEM, RENAMA, Campinas, Brazil, 2- than observed in humans. This is expected due to PluriCell Biotech, São Paulo, Brazil 3- Universidade CaCo2 thigh junctions. Main APAP hepatic Estadual de Campinas – Instituto de Biologia metabolites have not been found for the time being, but we were able to see a reduction in the APAP amount, The use of human primary cardiomyocytes (hCM) for showing HepaRG metabolic capability. These data cardiotoxicity depends on availability and have poor show that 2-OC is able to emulate some PK functions consistency. Cardiac cell lines phenotype differs from and can help on system’s optimization. primary hCM. Pluripotent human steam cells References:Adv Exp Med Biol. 2016;856:299-316. differentiated into cardiomyocytes (HiPSC-CM) are ALTEX. 2016;33(3):272-321.ALTEX 2015;32(4):327- available and have functional and morphological 78.Lab Chip. 2015 Jun 21;15(12):2688-99. J Pharm properties closer to hCM. Most groups report HiPSC- Biomed Anal. 2002 Jul 20;29(4):701-14. Eur J Pharm CM survival for no longer than 14 days. We produced Biopharm. 2015 Sep;95(Pt A):77-87. HiPSC-CM spheroid aggregates of 300 to 600 μM and Financial support / Acknowledgments: CNPq. keep them alive and continuously beating at 30 to 40 bpm for 70 days in the 2-OC (by TissUse GmbH). At the 70th day, the still beating heart organoids were AM 31 - AYAHUASCA EXPOSURE INDUCES fixed and stained with OsO4. Electron microscopy SIMILAR NEUROBEHAVIORAL RESPONSES showed functional sarcomeres and mitochondria in the IN ZEBRAFISH AND RODENTS. HiPSC-CM from organoids. X-ray 3D morphology (synchrotron micro-tomography) showed a ANDRADE, T.S. 1; JUNIOR, W.M. 2; OLIVEIRA, R. homogeneous cellular distribution with few 3; SANTOS, A.F.A. 4; LOPES, M. 1; MORAIS, J.A. discontinuities and few necrotic centers in the core. 2; GRISOLIA, C.K. 1; DOMINGUES, I. 5; CALDAS, These results show that microfluidic chips may E.D. 4; PIC-TAYLOR, A. 2 improve the functionality and survival of HiPSC-CM organoids. References: Biochimica et Biophysica Acta 1.Laboratory of Toxicological Genetics, Institute of 1863 (2016) 1728–1748. Scientific Reports 5:14088 Biology, University of Brasilia, Brasilia-DF, Brazil; 2. (2015) DOI: 10.1038/srep14088. Altern Lab Anim. Laboratory of Embryology and Development Biology, 2012 Oct;40(5):235-57 Review. Institute of Biology, University of Brasilia, Brasilia- Financial support / Acknowledgments: CNPq. DF, Brazil; 3. Laboratory of Ecotoxicology and Environmental Biology Profº Dr. Abílio Lopes (LEAL) and PPG Toxicology and Toxicological Analysis FCF/USP, University of Campinas, Faculty of Technology UNICAMP, 13484-332, Limeira, São ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 33

Paulo, Brasil; 4. Laboratory of Toxicology, Faculty of AM 32 - IN SILICO TOXICOPHORICAL Health Sciences, University of Brasilia, Brasilia-DF, ANALYSIS OF HIBALACTONE: A LIGNAN Brazil; 5. Department of Biology & CESAM, ISOLATED FROM HYDROCOTYLE University of Aveiro, Campus Universitário de UMBELLATA L. (ACARIÇOBA) – Santiago, Aveiro, Portugal. ARALIACEAE

Ayahuasca is a psychoactive concoction prepared with OLIVEIRA, T. L. S. 1,4; OLIVEIRA, M. G. 2; the plants Banisteriopsis caapi and Psychotria viridis SILVA, V. B. 3; MORAIS, S. R. 1,4; COSTA, E. A. 2; and used ancestrally by Amazonian indian populations, PAULA, J. R. 5 and more recently, by religions in Brazil and other countries. This sacred brew has healing properties and 1 Faculdade de Farmácia da Universidade Federal de is considered to be safe in ritualistic context. However, Goiás, Goiânia – GO, Brasil; 2. Instituto de Biologia the prospect of ayahuasca therapeutic use still lacks of da Universidade Federal de Goiás, Goiânia – GO, preclinical data to certify its safety. This study aimed Brasil; 3. Escola de Ciências Médicas, Farmacêuticas e at identifying the ayahuasca effects in early fish Biomédicas da Pontifícia Universidade Católica de development and compares its neurobehavioral effects Goiás, Goiânia – GO, Brasil; 4. Universidade Paulista in zebrafish embryos and rat models. The infusion was – UNIP, Goiânia – GO, Brasil. provided by the religious group União do Vegetal and was previously characterized. Toxicity and Introduction: In silico toxicophorical analysis is one developmental endpoints for zebrafish embryos were type of toxicity assessment that uses computational assessed at 0 to 1000 mg/L during 96 h of exposure. resources instead of animal models as well as to guide The effects on locomotor activity of zebrafish larvae experimental in vivo assays. Hydrocotyle umbellata L. were assessed using the tracking system Zebrabox at 0 have been used in folk medicine, however scientific to 20 mg/L and after 120 and 144 h of exposure. The toxicological studies in support of this medicinal use ayahuasca infusion was administered once by gavage and description of the chemical composition of the to Wistar rats at 1, 5 and 15X the dose taken during species are not clarified. Objective: This study aimed religious rituals, and neurobehavioral effects evaluated the investigate the in silico toxicological potential of after 2 h in the open field (OF), elevated plus-maze hibalactone, a lignan isolated from H. umbellata, in (EPM) and forced swimming (FS) apparatus. The order to screen potential hazards. Materials and LC50 of ayahuasca in zebrafish was estimated to be Methods: The hibalactone was isolated from the 236.3 mg/L. Ayahuasca caused a series of ethanolic extract prepared with underground parts of developmental anomalies in zebrafish embryos, mainly the plant by column chromatography (CC). Structure at the highest concentration tested, including hatching of the isolated compound was elucidated by delay, loss of equilibrium, edema and accumulation of spectroscopic techniques 1H and 13C RMN and mass red blood cell. The behavior of embryos exposed to the spectra (MS), further confirmed by X-ray highest concentration was also affected with a decrease crystallographic. Toxicity prediction of hibalactone 2D in locomotor activity. Decreased locomotion was also structure was performed with ACD/I-Lab, Lazar, Pred- observed in rats treated at the highest dose in the OF hERG and Pred-Skin online tools, which are based on and EPM apparatus, and a higher swimming time in chemical similarity criteria with known toxic the FC, suggesting a possible antidepressant effect. compounds. Results: A lignan was first time isolated in These results indicated that the effects of ayahuasca species of H. umbellata in fraction dichloromethane correlated well for zebrafish embryos and rodents, obtained from the ethanolic extract. The chemical showing that zebrafish may provide a useful model to structure of the isolated compound was determined by study ayahuasca and other hallucinogenic drugs. NMR spectroscopic data (1H and 13C NMR, HMQC Further research focusing on the molecular pathways and HMBC), mass spectra (MS) and x-ray affected by ayahuasca administration in both zebrafish crystallography. The data suggested that the structure and rat could provide more information on the refers to lignan dibenzil butirolactone, hibalactone. potential of ayahuasca as an antidepressant. Prediction data of hibalactone revealed no Financial support / Acknowledgments: The present toxicological potential regarding Ames toxicity, hERG study received financial support of the Federal District inhibition and binding to the estrogen receptor. In Research Foundation (FAP-DF) within the post- contrast, a risk of carcinogenicity and skin sensitisation doctoral grant (1250/2016) to Andrade T.S., and the were detected. Furthermore, the analysis of adverse grant to Junior, W.M. Grisolia, C.K., is grateful to effects on a number of organs and their systems within CNPq for the “Bolsas de Produtividade”. Oliveira, R., the therapeutic dose range revealed that hibalactone acknowledge the Brazilian Ministry of Education, and present high probability to cause adverse effects on Ministry of Science and Technology of Brazil for the blood, cardiovascular system, gastrointestinal system, scholarship provided through the program Science kidney, liver and lungs. Conclusion: This study without Borders (CNPq BJT-A). contribute to knowledge of toxicological potential of ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 34 the hibalactone isolated in species of H. umbellata and induce the toxic property, helping scientists to propose indicates that this compound could be studied in vivo structural modifications to increase the safety of by animal model. desired compounds. Moreover, the Pred-Skin app has Financial support / Acknowledgments: CNPq, been also developed for mobile devices and is CAPES, FAPEG. available at the App Store. Therefore, the developed tools ensure both highly accurate predictions and easy interpretation of the models, allowing users to AM 33 - DEVELOPMENT OF INNOVATIVE IN discriminate potential toxicants for hERG liability and SILICO PLATFORMS FOR EARLY TOXICITY human skin sensitization as well as to purpose PREDICTION OF CHEMICALS structural modifications. Financial support / Acknowledgments: CNPq, ALVES, V. M. 1,2; BRAGA, R. C. 1; MURATOV, E. CAPES, FAPEG, NIH. N. 1,2; STRICKLAND, J. 3; KLEINSTREUER, N. 4; TROPSHA, A. 2; ANDRADE, C. H. 1.

1 Laboratory for Molecular Modeling and Drug AM 34 - STRATEGY COMBINING NON- Design, Faculty of Pharmacy, Federal University of ANIMAL METHODS FOR EVALUATING Goiás, Goiânia, GO, 74605-170, Brazil; 2 Laboratory OCULAR TOXICITY OF AGROCHEMICALS for Molecular Modeling, Division of Chemical FORMULATIONS IN BRAZIL Biology and Medicinal Chemistry, UNC Eshelman School of Pharmacy, University of North Carolina, ANDRADE, W. A. 1; SILVA, A. C. G. 1; MOREIRA, Chapel Hill, NC, 27599, USA; 3 Integrated Laboratory L. C. 1; GOMES, T. R. L. S. 1; CARVALHO, A. B. 1; Systems, Inc., P.O. Box 13501, Research Triangle VALADARES, M. C. 1 Park, NC, 27709, USA; 4 National Toxicology Program Interagency Center for the Evaluation of 1 Universidade Federal de Goiás, Goiânia – GO, Alternative Toxicological Methods, National Institute Brasil. of Environmental Health Sciences, Research Triangle Park, NC, USA. Introduction: different strategies for classification and labelling (C&L) of ingredients/mixtureshave been Testing of chemicals on animals has been publicly and considered by Regulatory Agencies.Regarding to politically less tolerated worldwide, and testing of agrochemicals, the lack of global harmonization for cosmetic ingredients is completely prohibited in C&L isthe main problem for the implementation of Europe and in many other countries, including some alternative methods. On the other hand, in vitro states in Brazil. Consequently, there is a strong need methods have been successfully adopted to assess the for developing novel alternative methods to better potential of toxicity of chemicals. Objective: assess toxicity while avoiding or reducing animal development of a strategy combining non-animal testing. In silico or computational tools have been methods for ocular toxicity of agrochemicals. Material recognized as a practical solution to evaluate and Methods: 86 agrochemicals from different ocular substances not yet experimentally tested, providing toxicity categorization were selected. As a first step, speed, low cost, and high-quality predictions during the potential for ocular toxicity of agrochemicalswere the early stages of drug/cosmetic development or evaluated using the Short Time Exposure (STE) test. environmental toxicity assessment. In this work, we In the second step, the samples not classified and non- will present the development of two innovative and irritating by STE were evaluated by the Bovine freely accessible web-based and mobile applications Corneal Opacity and Permeability (BCOP) test for identification of putative hERG blockers and followed by histomorphometric evaluation. The third human skin sensitizers, as the first step of a multi- step was to assess the potential of induction of vascular tiered testing strategy with higher predictivity than the damage, by the Hen's Egg Test-Chorioallantoic animal testing. The applications are based on robust Membrane (HET-CAM) assay (low false negative). and predictive Quantitative Structure-Activity Results: From the 86 agrochemicals, the STE Relationships (QSAR) models developed using the categorized 73 samples, 68 as irritant (category 1) and largest publicly available datasets of structurally 5 as non-irritant (no category); and 13 samples could diverse compounds including a variety of drug classes not be classified. Considering that the STE has false of and chemical compounds. The freely available Pred- 1% and a false negative of 51%, when compared to the hERG and Pred-Skin platforms rabbit eye test, these 18 samples wereinvestigated by (http://labmol.com.br/predherg and BCOP test. Excluding alcohols and solids, BCOP has a http://labmol.com.br/predskin) combine in silico false positive of 20% and a false negative of 8%. The toxicity prediction with a visualization output, which BCOP test categorized 13 samples as non-irritant. The indicates which structural alerts are more likely to 5 agrochemicals unclassified by conventional BCOP, ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 35 were histomorphometrically analyzed and tissue damage were observed, then being categorized as irritant and confirmed using the HET-CAM assay. In addition, we observed that some agrochemical formulations with the same active compound and same concentration showed different toxicological categorization. Conclusion: Since C&L of agrochemicals in Brazil is not based on the GHS, this tier platform, using a Top-Down approach, could be useful to harmonize the categorization of agrochemicals in Brazil, providing regulatory subsidies for ANVISA. Financial support / Acknowledgments: FAPEG / CAPES / CNPQ / FUNAPE

ANALYTICAL TOXICOLOGY ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 37

AN 01 - DEVELOPMENT OF ANALYTICAL AN 02 - ANTINEOPLASTIC PROTOTYPE METHOD FOR IDENTIFICATION OF BRUCIN, DRUG LQFM018: STUDY OF THE STRICHININE AND METHOMYL IN GASTRIC PHARMACOKINETIC AND ALLOMETRIC CONTENT OF ANIMALS BY THIN LAYER PROFILE BY LC-MS/MS CHROMATOGRAPHY RODRIGUES, A. R.1, RODRIGUES, C. R.1, BAIRROS, A. V. 1; DIAS, D. O. 1 GOMES, S. A.1, ZOGHAIB, A. F.1, ZOGHAIB, I. V.1, OLIVEIRA, F. M.1, CUNHA, L. C.1 1 Universidade Federal de Santa Maria 1. Núcleo de Estudos e Pesquisas Tóxico- This work describes the development of a qualitative Farmacológicas – NEPET, Faculdade de Farmácia, analytical method for brucine, strychnine and Universidade Federal de Goiás – UFG, Goiania-GO, methomyl by thin layer chromatography (TLC) in Brasil. gastric content. Optimization of the method included studies of TLC plate size, type of mobile phase, pH Introduction: LQFM018 is a candidate to prototype values, volume of solvents, time of agitation and antitumor drug obtained by molecular simplification number of extraction step to isolate the analytes. Ten from anticancer compounds called nutlins, inhibitors centimeters of silica gel was used as the stationary of the p53-MDM2 interaction. Goal: This study aimed phase and chloroform: ethyl acetate: diethylamine (0.5: to develop and validate a bioanalytical method for 8.5: 1) as the mobile phase. A liquid-liquid extraction quantification of LQFM018 in plasma and study its at pH 12 with a volume of 15 ml of chloroform: ethyl pharmacokinetic. Materials and Methods: Validation ether (2:1) during 2 minutes of agitation using 5 mL of contemplated the following tests: linearity, precision, sample. After, the solvent was evaporated and diluted accuracy, selectivity, recovery, matrix effect, residual with 500 µL of methanol. This volume was passed to effect and stability. The validated bioanalytical method another bottle, dried, resuspended with 50 µL of used the following analytical parameters: ACE® C18 methanol and applied in TLC plate. It was used column (100 mm × 4.6 mm, 5 µm); mobile phase: Dragendorff reagent for chemical revelation. Retention buffer 2 mM ammonium acetate with 0.025% formic factor (Rf) for brucine, strychnine and methomyl was acid and methanol (50%:50% v/v); flow: 1.2 mL/min; 0.3, 0.37 and 0.6 respectively. This method was internal standard (IS): domperidone; liquid-liquid validated according the Guidance for the Validation of extraction with methyl tert-butyl ether (MTBE) and Analytical Methodology for TLC from UNODC which injection volume of 3.0 µL. LQFM018 was limit of detection (LD), specificity and selectivity, administered to 3 females Wistar rats at 100 mg/kg, precision and reproducibility were performed. LD was i.p. After administration, 0.5 mL samples of blood established with the analysis of 10 independent were collected by cannulation of the left jugular vein samples at the lowest concentration detected. LD of with heparinized syringe. Results and Discussion: The brucine, strychnine and methomyl was 3 µg/mL, 30 method was linear from 10 to 15,000 ng/mL, intrarun µg/mL and 500 µg/mL respectively. Specificity and precision was ranged from 0.6% to 5.5%, interrun selectivity were performed using tetramisole, from 1.8% to 6.7%, accuracy from 99.0% to 107.0% ibuprofen, salicylic acid, scopolamine, dipyrone, and recovery of 74.1% ± 4.9%. Retention times were warfarin, phenobarbital, diazepam and paracetamol; 10 3.16 min (LQFM018) and 1.81 min (IS). different blank samples were tested. No interference Pharmacokinetic parameters (mean ± SD) were: t½ = was observed in these tests. For precision and 2.89 h ± 2.0; ClT/F = 22.01 mL/min/kg ± 13.5; Vd/F = reproducibility, the analysis were performed on 5.48 L/kg ± 3.6. To Vd value, applying the allometric different days (n = 5) with different operators (n = 2) scaling for an adult weighing 70 kg, a value of 1954.3 considering LD of the analytes in these tests. The L/kg is found, which suggests extravascular method developed and validated in TLC has proved to accumulation of the compound. In allometric scaling, be reliable and reproducible and can be used as the value of half-life correspond to 12.6 h. Total screening in suspected cases of intoxicated animals clearance value found resulting in a value of 1800.4 and to help in the treatment of these cases, reducing mL/min for a person weighing 70 kg, indicating waiting for toxicological reports and costs. greater participation of hepatic elimination Financial support / Acknowledgments: FIEX mechanisms once the fact that the fat-soluble molecule of the compound requires a prior biotransformation to increase its polarity so that it can be eliminated. Conclusion: Results, extrapolated to humans, revealed large half-life, high Vd and high CLT, allowing the comprehension that the prototype showed good tissue distribution profile and was extensively eliminated.

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 38

Financial support / Acknowledgments: CAPES; butyrylcholinesterase activity and methemoglobinemia NEPET UFG; INSTITUTO DE CIÊNCIAS represented 11%, 10% and 0.25% of total analyzes, FARMACÊUTICAS-ICF respectively. Among the new techniques implemented, AChE corresponded to 2.8% and SAL 1.4% of the total analyzes. Conclusion: There was a considerable AN 03 - PROFILE OF THE TOXICOLOGICAL increase in the demand after 2 years of operation, ANALYSES AFTER TWO YEARS OF however the requests profile remained similar, despite LABORATORY OPERATION the new techniques implemented. The DAS remained the main request. SILVA, C.I.1; RAMOS, V.B1; PACHECO, L.K..1; Financial support / Acknowledgments: Universidade KRETZER, S.L.1; MAIDANCHEN, T.1 Federal de Santa Catarina VENDRUSCOLO,D.T.1; VANDRESEN, B.1; NASCIMENTO, A.L.1; MATIOLLO, C.1; DASSOLER, F.J.1; DOS SANTOS, C.R.1,2 AN 04 - A NOVEL PRECURSOR ION SCAN METHOD FOR NBOME EVALUATION IN 1.Divisão de Análises Clínicas, Hospital Universitário WHOLE BLOOD Professor Polydoro Ernani de São Thiago, Universidade Federal de Santa Catarina, Florianópolis- MORAIS, D. R. 1; CUNHA, K. F. 2,3; EBERLIN, M. SC, Brasil; 2.Departamento de Patologia, Centro de N. 1; COSTA, J. L. 2,3 Ciências da Saúde, Universidade Federal de Santa Catarina, Florianópolis-SC, Brasil. 1. Laboratório ThoMSon de Espectrometria de Massas - Instituto de Química, Universidade Estadual de Introduction: Toxicological analyzes assist in the Campinas, Campinas - SP, Brasil; 2. Centro de diagnosis and medical management for acute Controle de Intoxicações de Campinas - Faculdade de intoxications.Despite the recognition of their Ciências Médicas, Universidade Estadual de importance, their realization depends on the substances Campinas, Campinas - SP, Brasil; 3. Faculdade de that can be evaluated, as well as on the awareness of Ciências Farmacêuticas, Universidade Estadual de the teams involved in the implementation of the Campinas, Campinas - SP, Brasil. requested analyzes.Objective:To compare the profile of the toxicological analyzes performed in the NBOMes are highly potent novel psychoactive Toxicology Laboratory (TL) at University substances (NPS) that causes intense hallucination, Hospital/UFSC in the first 10 months with the same tachycardia, hypertension and aggressiveness at low period after 2 years of official operation.Methodology: doses. Previous studies usually analyze NBOMes in Retrospective descriptive study based on the requests biological fluids using target methods based on liquid of toxicological analyzes and samples records sent to chromatography-tandem mass spectrometry (LC- the TL in the periods from September/2014 to MS/MS) in multiple reactions monitoring (MRM) June/2015 (P1) and September/2016 to June/2017 mode, however, there are no screening methods for (P2).Results: After two years of operation, two new NBOMes in biological fluids. Precursor ion scan (PIS) quantitatives methodologies were implemented: is a very popular technique to study classes of salicylemia (SAL) and acetylcholinesterase activity compounds, which have a diagnostic fragment ion, (ACHE), both by spectrophotometry. While in P1 a such as NBOMe class. The aim of this work was to total of 422 analyzes were performed, in P2 this develop a screening method for NBOMe compounds number increased 87% (789 analyzes), representing an in whole blood, using LC-MS/MS. The sample average increase of 37 analyzes/month. There was a extraction was performed with 300µL of whole blood, slight increase in qualitative determinations 300 µL of a saturated solution of sodium tetraborate (immunocromatography and colorimetry) (67.3% in P1 and 1.2 mL of methyl tert-butyl ether. The mixture was and 73.8% in P2) that occurred due to the large vortexed for 5 min at 2,000rpm, centrifuged at 8,000 number of drug abuse screening tests (DAS). rpm for 5 min, 900 µL of organic phase was However, the distribution of the detected results for transferred to a clean tube, and then the solvent was DAS in P2 (723) remained similar to P1: the evaporated. The sample was resuspended with 500 µL benzodiazepine group was the most frequently of methanol, transferred to an autosampler vial and 20 detected (30.8%), followed by Cocaine (21%) and µL were injected into the LC-MS/MS system (Agilent Marijuana (11%). The paraquat research represented 1260 Infinity LC system coupled to an ABSciex 5500 3.2% of total analyses and was detected in 48% of the QTRAP mass spectrometer). The chromatographic cases. The quantitative analyzes were performed by separation was performed with a C18 column (Atlantis automated method (Dimension®rxl- T3, 150x3 mm, 3 μm, Waters®), the mobile phase Siemens,Germany) and spectrophotometry. Among consisted of water (A) and acetonitrile (B), both of them, the plasma concentration of paracetamol, containing formic acid (0.1%, v/v), gradient elution ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 39 and flow rate of 0.50 mL/min. The mass spectrometer one was the implantation of the method for the was set to operate with an electrospray source, in analysis of real samples from patient under treatment positive ion mode, using PIS as survey scan – for chemical dependence and the comparison between compounds which presented the NBOMe class specific this method and the High Performance Thin Layer fragment, m/z 121 (collision ion spread, 35 eV± 15 Chromatography (HPTLC) method, which was the eV). The compounds were identified by comparison actual confirmatory method used at the NAL-CIT/RS. with an in house MS/MS spectrum library. The Results indicate that the liquid-liquid extraction is the developed method was able to detect several NBOMes, best extraction method, that N,O- e.g. 25H-, 25D-, 25G-, 25E-, 25C-, 25I- and 25B- Bis(trimethylsilyl)trifluoroacetamide (BSTFA) is the NBOMe in concentrations lower than 100 pg/mL. PIS best derivatization agent and that the GC-MS method mode using the specific NBOMe class compound is the most sensitive, specific, rapid, free of toxic fragment, m/z 121, in tandem with enhanced product residues when compared to the HPTLC method, ion scan mode is a great selectivity method and robust allowing more reliable results, thus avoiding approach for rapid untargeted NBOMe screening, unnecessary expenses with prolonged or reduction which leads to identification of NBOMe structure. rehabilitation. Financial support / Acknowledgments: The authors Financial support / Acknowledgments: Apoio thank FAPESP (process number 2015/10650‐ 8 and Centro de Informação Toxicológica do Rio Grande do 2016/23157‐ 0), CAPES (process number Sul (CIT/RS). 23038.006844/2014‐ 46) and CNPq (process number 830525/1999‐ 8), by the fellowships and the financial support. AN 06 - LIQUID CHROMATOGRAPHY– TANDEM MASS SPECTROMETRY APPLIED TO THE STUDY OF COCAINE AND AN 05 - DEVELOPMENT AND APPLICATION METABOLITES IN DRIED URINE SPOTS (DUS) OF A METHOD FOR DETECTION OF 11-NOR- 9-CARBOXY-Δ9-THC (MARIJUANA BARBOSA, I.L.1; COSTA, J.L 1; EBERLIN, M. N. 1 BIOMARKER) IN HUMAN URINE BY GAS CHROMATOGRAPHY COUPLED TO MASS 1. Universidade Estadual de Campinas SPECTROMETRY Dried blood spots (DBS) provides several advantages DALLEGRAVE, E.1; FRAGA, M.P. 1; CHRIST, T.S. over conventional sample storage methods, mainly in 1; SEBBEN, V.C.2;TSAO, M.1. what it concerns transport and stability of the analytes and it has been used for clinical and forensic 1 Universidade Federal de Ciências da Saúde de Porto applications. Similarly, it is possible to perform Alegre, Porto Alegre - RS, Brasil; 2 Centro de toxicological analysis of drugs of abuse in samples of Informação Toxicológica do Rio Grande do Sul, Porto dried urine spots (DUS) and it should be of interest Alegre - RS, Brasil. supposing that shares the same advantages as DBS. In this work, a very simple analytical method for DUS Considering that the most used form of Cannabis and LC-MS/MS was developed for the screening and sativa, as an illicit drug all over the Americas, is the confirmation of cocaine and its metabolites: marijuana and that confirmatory methodologies benzoylecgonine (BZE), ecgonine methyl ester (EME) capable of detecting the 11-nor-9-carboxy-Δ9- and cocaethylene (EC). To prepare DUS cards, spiked tetrahydrocannabinol (THC-COOH) are extremely urine samples (20 µL) were spotted onto Whatman 903 important for drug testing at the work place, in the Protein Saver cards followed by 2 hours air-drying control of doping in sports, for the follow up of prior to extraction. The extraction was performed using patients under clinical rehabilitation and for forensic 300 µL of methanol:water (60:40) with 0,1% formic purposes, this study aimed to develop a confirmatory acid, containing the internal standard (cocaine-d3, 10 method for the detection of the THC-COOH ppb) followed by 15 minutes of vortex and 5 minutes metabolite of marijuana by Gas Chromatography of centrifugation at 10.000 rpm. After this process, 200 coupled to Mass Spectrometry (GC-MS) to be used in µL of supernatant was transferred to a vial and 15 μL the Nucleus of Laboratorial Analysis of the injected on a UPLC-MS/MS system (8040 Shimadzu, Toxicological Information Center of Rio Grande Japan). Separation was performed in a C18 column (NAL-CIT / RS). The study was carried out in two (Atlantis T3, 150 x 3 mm, 3 μm, Waters®) with a phases: the first one for the development of the gradient elution with a flow rate of 0.40 mL/min. method, which evaluated the best methodology for the Chromatography separation was carried out using extraction of the metabolite THC-COOH, allowed the ammonium formate 2 mM and 0,1% of formic acid in evaluation of the best derivatization agent and the (A) water (B) methanol in a 16 minute run. Linearity determination of the method’s cutoff level; the second was obtained ranging from 25 (LOQ) to 1,000 ng/mL ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 40 for the four compounds (r > 0.99, 1/x weighted linear with temperature at 180°C, raised to 270°C (35°C regression). Validation for this method is following the ⁄min) and to 310°C (20°C⁄min) for 22 min, transfer line recommendations from Scientific Working Group for 300°C and quantification ion 573 m/z. Results: The Forensic Toxicology (SWGTOX) for quantitative tert-buthyldimethylsilyl ether areas ranged from 3,067- analysis. Precision (CV (%)) and bias (%) were 78,094. The response was modulate in a quadratic evaluated within and between runs at 3 different levels equation with the factors and its interactions, (75, 400 and 800 ng/mL), never exceeding 20% at presenting significant adequate adjustment (P0.05). By each concentration. Matrix effect and dilution integrity means of the equation: Area= -2,0x107 precision and bias criteria were also tested and met. +2,1x106*time+2,8x105*conc.+3,9x105*temp.+2.260 Stability is being study for a period of 0,7,15,30,60 and *time*conc. –15.691*time*temp. +223*conc.*temp.– 90 days. Since this method is so simple and the use of 2,1x105*time2 –1698* conc.2 -2228*temp.2, the dried sample has so many advantages, it is expected optimum reaction condition was set as: 3 h, that in the future it can be used in forensic toxicology derivatizant at 16% and temperature 50ºC. Conclusion: routine. It was possible to define optimal conditions for the Financial support / Acknowledgments: CAPES derivatization reaction within a short period of time, that can be used in the development of a GC-MS method for the quantification of 4β-OHC. AN 07 - USE OF SURFACE RESPONSE Financial support / Acknowledgments: FAPERGS; METHODOLOGY IN THE OPTIMIZATION OF Universidade Feevale THE EXPERIMENTAL CONDITIONS OF 4β- HYDROXYCHOLESTEROL DERIVATIZATION WITH BUTHYL DIMETHYLSILYL AN 08 - METHOD VALIDATION FOR IMIDAZOLE IN GC-MS ANALYSIS ACTIVITY SIMULTANEOUS QUANTITATION OF HIPPURIC ACID AND 3-METHYLHIPPURIC MANFRO, I. D. 1; TEGNER, M. 1; RAYMUNDO, S. ACID IN URINE 1; MULLER, V. V. 1; HAHN, R. Z. 1; LINDEN, R. 1; ANTUNES, M. V. 1 FUSINATO, M.N.1, NASCIMENTO A.L.1, DOS SANTOS, C.R.1,2., SILVA, C.I.1 1 Laboratório de Toxicologia, Centro Integrado de Especialidades em Saúde, Universidade Feevale, Novo 1.Divisão de Análises Clínicas, Hospital Universitário Hamburgo-RS, Brasil Professor Polydoro Ernani de São Thiago Universidade Federal de Santa Catarina, Florianópolis- Introduction: 4β-hydroxycholesterol (4β-OHC) is a SC, Brasil; 2. Departamento de Patologia, Centro de metabolic product of cholesterol formed by the Ciências da Saúde, Universidade Federal de Santa CYP3A enzyme, being a potential endogenous Catarina, Florianópolis- SC, Brasil phenotyping biomarker. 4β-OHC analysis by GC-MS after derivatization was previous described, but it can Introduction: Toluene and xylene are oil derivatives be time-consuming and expensive. Considering the widely used in the industry as solvents used in paints influence of multiple factors into the reaction yield, the and adhesives. Due to neurotoxic and nephrotoxic experimental conditions of derivatization must be action, biological monitoring using the hippuric acid carefully assessed. The response surface methodology (HA) and 3-methylhippuric acid (3-MH) is is a set of statistics techniques employed to model and recommended to evaluate occupational exposure to analyze problems in which a response variable is toluene and xylene. Since the co-exposure is frequent influenced by diverse operational variables. Objective: the determination of these compounds simultaneously To optimize the experimental conditions for the 4β- allows reduce the time and cost of the analysis. OHC derivatization reaction with buthyl dimethylsilyl Objective:Validate a High Performance Liquid imidazole (TBDMSIM) to a tert-buthyldimethylsilyl Cromatography Photodiode Array Detector method ether. Material and methods: A Box-Behnken (HPLC-DAD) for AH and 3-MH determination in experimental design was set with the variables: urine samples. Materials and methods:Calibration incubation temperature (25; 50; 75ºC), concentration samples were prepared in urine to obtain 5 calibrations of TBDMSIM (8; 14; 20%) and incubation time (1; levels containing HA and 3-MH (0.312; 0.625; 1.25; 3,5; 6 hours) in a total of 17 runs. A 4β-OHC 100 2.5 and 5.0 mg.mL-1). The samples (1.0 mL) were ng⁄ml solution was analyzed through different extracted using 1.0 mL of methanol, followed of combinations after adding 100 µL of the TBDMSIM centrifugation at 3,000 rpm for 7 minutes. solution. After, it was extracted with MTBE and the Chromatographic separation was performed on a C-18 organic phase dried at 45 ºC. The extract was reverse phase column (4.6 mm x 15 cm x 5 μm), using resuspended and 1 µL was injected into a GC-MS, acetic acid 0.8%:methanol (80:20) as mobile phase, with a HP5MS column (30m x 0.250mm, 0.25µm) flow rate at 0.4 mL.min-1; column oven at 40ºC and ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 41 injection of 10 μL of supernatant in the HPLC-DAD precision, accuracy, specificity, robustness, and with detection at 230 nm. For validation, the following stability were evaluated. Method applicability was parameters were evaluated: Limit of Detection (LOD), proved by the MPH analysis in sewage. Results: MPH Limit of Quantification (LOQ), Linearity, Intra e Inter- retention time was 7.2 min. A linear correlation days Precision and Stability, according to the RE between MPH concentration and detector response was 899/03 ANVISA.Results:The method demonstrated achieved in the range of 10 to 125 µg/mL. Residuals good separation with retention times of 4.24 min ± showed a normal distribution, independence, and 0.13 (HA) and 10.59 min ± 0.23 (3-MH) and linearity homoscedasticity. LD and LQ were 0.27 and 0.92 in the ranges from 0.312 to 5.0 mg.mL-1 for both µg/mL, respectively, indicating the sensitivity of the analytes (r² > 0.99). It presented good sensitivity with method. Precision and accuracy results show an LOD of 0.404 and 1.69 μg.mL-1 and LOQ of 1.22 e acceptable range. None of the factors studied had a 3.84 μg.mL-1 for HA and 3-MH, respectively. In the significant effect on the quantification of the MPH intra and inter-days precision, the relative standard demonstrating the robustness of the method. MPH deviation were lower than 3.0% and 3.5%, solutions demonstrate to be stable for at least seven respectively. Samples were considered stable when days when kept at room temperature. The recovery of kept at room temperature for 2 days, under MPH after SPE step was 100%. Through this method, refrigeration for 15 days and under freezing for 90 it was possible to found 2.8 µg/L MPH in sewage. days, supporting up to 3 cycles of thawing. Conclusion: The proposed LC-PDA method can be Conclusion: The results obtained shows that it can be regarded as selective, accurate, precise, and valid for used for the biological monitoring of occupational determination of MPH water contamination. exposure to these solvents in compliance with the provisions of current legislation in Brazil. Financial support / Acknowledgments: Hospital AN 10 - BIOCHEMICAL PARAMETERS Universitário UFSC e Ministério Público do Trabalho ROUTINELY APPLIED IN ECOTOXICOLOGY de Santa Catarina VARY SIGNIFICANTLY WHEN DETERMINED IN WET AND FREEZE-DRIED ANIMAL TISSUES AN 09 - DEVELOPMENT AND VALIDATION OF A LC-PDA METHOD FOR DABKIEWICZ, V. E. 1; CARVALHO, L. V. B. 1; METHYLPHENIDATE WASTEWATER SENNA, M. M. 1; MOREIRA, J. C. 1; HAUSER- ANALYSIS DAVIS, R. A. 1

FREDDO, N. 1; PAIXÃO, M. C. S. 1; BERTOL, C. 1Centro de Estudos da Saúde do Trabalhador e D. 1; GRANDO, L. G. R. 1 Ecologia Humana, Escola Nacional de Saúde Pública Sergio Arouca, Fundação Oswaldo Cruz, Rio de 1 Universidade de Passo Fundo, Passo Fundo - RS, Janeiro – RJ, Brazil. Brasil Introduction: Biochemical determinations commonly Introduction: Environmental contamination by used to assess effects of environmental contamination, medicinal residues are often reported. These chemicals mainly, oxidative stress, in ecotoxicological assays are are not monitored or removed in water treatment routinely conducted in both wet and freeze-dried (dry) stations and they are considered emerged pollutants. tissues in sentinel organisms, like fish. However, Objective: To develop and validate a feasible method differences in concentrations between these tissues for methylphenidate (MPH) waste analysis using high occur, and oxidation of certain parameters is a concern. performed liquid chromatography coupled with Objectives: Compare three biochemical parameters, photodiode array ultraviolet detector (LC-PDA). metallothionein (MT), reduced glutathione (GSH) and Material and methods: LC analysis was performed lipid peroxidation (LPO), in dry and wet fish samples. using a Flexar Perkin Elmer liquid chromatograph Material and methods: Tilapia livers (n=5) and muscle equipped with a Flexar PDA detector and a Flexar LC (n=4) from a market in Rio de Janeiro were split into autosampler. Chromatographic separation was two halves, one freeze-dried for 48 h and the other achieved using a Perkin C18 reverse phase column frozen at -80o C. MT was purified by thermal (250 mm x 4.6 mm, 5 µm). The mobile phase extraction. Tissues were homogenized in Tris-HCl consisted of methanol: phosphoric acid 0.025% buffer, pH 8.6, with 0.5 mM phenylmethylsulfonyl (80:20) with a final pH adjusted to 5.9 with fluoride and 0.01 % β-mercaptoethanol and triethylamine. Flow rate, injection volume and centrifuged at 20,000g (60min; 4°C). The supernatants wavelength were 1 mL min-1, 20 µL and 206 nm, were heated (70°C; 10min), centrifuged again at respectively, and the run time was 10 min. Linearity, 20,000g (30min; 4°C) and frozen. For GSH, samples limits of detection (LD) and quantification (LQ), were homogenized in 0.1 M, pH 7.0, Na2HPO4 buffer ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 42 and 0.25 mM sucrose, centrifuged at 11.000g (30min; AN 12 - DEVELOPMENT AND VALIDATION OF 4°C) and frozen. For LPO, samples were homogenized A METHOD FOR THE SIMULTANEOUS in RIPA buffer, centrifuged at 1,600g (10min; 4oC) DETERMINATION OF IRINOTECAN AND and frozen. MT and GSH were determined by METABOLITES IN HUMAN PLASMA BY Ellman’s spectrophotometric reaction and LPO by a LIQUID CHROMATOGRAPHY ASSOCIATED commercial kit via malondialdehyde determination. WITH FLUORESCENCE DETECTOR (HPLC- Results: After correcting dry results for water loss FL) (70%), significant differences (p0.05). Conclusions: Lower MT and GSH levels in wet samples are HAHN, R. Z. 1; COSTA, P. A. 1; CEZIMBRA, A. 1; probably due to auto-oxidation, since wet samples ANDRIGUETTI, N. B. 1; SCHNEIDER, A. 1; REIS, suffer more freeze-thaw cycles than dry samples. S. L. 2; KLÜCK, H. 2; SCHWARTSMANN, G. 2; Regarding LPO, water removal may eliminate certain ANTUNES, M. V. 1; LINDEN, R. 1 pro-oxidants, like metal ions, and inhibit the formation of secondary lipid oxidation products. Thus, freeze- 1. Laboratório de Análises Toxicológicas, drying is more appropriate when determining these Universidade Feevale, Novo Hamburgo-RS, Brazil; 2. biochemical parameters, providing more reliable Universidade Federal de Ciências da Saúde de Porto results than wet samples. Alegre, Porto Alegre-RS, Brazil. Financial support / Acknowledgments: Fiocruz Background: Irinotecan (IRI) is a chemotherapeutic agent, used mainly in the treatment of colorectal and AN 11 - IMPLEMENTATION OF AUTOMATED advanced pancreatic cancer. An important limitation HEADSPACE TECHNIQUE BY GC / MS OF associated with the use of IRI is the significant THE ETHANOL BLOOD DOSE APPLIED TO variability in the therapeutic response and the TOXICOLOGY occurrence of toxicity. The active metabolite SN-38 is 100 to 1,000 times more cytotoxic than IRI, being PETERS, G. S. 1; FEISTEL, C. C. 1, 2; VIANNA, R. metabolized to its inactive glucuronide SN-38G. The L. 2; SEBBEN, V. C. 1, 2 availability of a sensitive method to quantify IRI and metabolites may allow the individualization of IRI 1. Universidade Luterana do Brasil, Canoas - RS, doses. Objective: To develop and validate a method for Brasil; 2. Centro de Informação Toxicológica / Centro the simultaneous determination of IRI and SN-38 in Estadual de Vigilância em Saúde - RS, Brasil; plasma and estimation of SN38G by HPLC-FL. Method: Aliquots of 200 μl plasma added with internal Analyzes of ethanol and its metabolites are of great standard (IS, camptothecin) were precipitated with 400 interest to the health system, not only because of its μL of acetonitrile:methanol mixture (1:1, v/v). After, potential for psychosocial transformation, but also 400 μL of the supernatant was extracted with 1 mL of because of the toxicity and metabolic disorders that methyl-tert-butyl ether. The separated organic phase this drug causes in the body. The present work was dried and recovered with 200 μl of the mixture of describes a validation as well as implementation of the mobile phase:HCl 1 M. For determination of SN-38G, technique for the detection and quantification of enzymatic hydrolysis with β-glucuronidase was ethanol in human blood using an automated headspace performed immediately before protein precipitation. method by Gas chromatography coupled to mass The concentration of SN-38G was determined by spectrometry. The study of the method is based on the subtraction of the molar equivalents. Fifty microliters volatility of the ethanol molecule, and the tests were of recovered extract was injected into the HPLC-FL, applied in glass containers, resistant and hermetically with separation in an Eclipse Plus C8 (150 × 4.6 mm, 5 sealed with a septum, where extraction under heating μm) column. Mobile phase was a mixture of 0.1 M was carried out by means of a modified syringe called phosphate buffer pH 4.0 and acetonitrile (80:20, v/v), gas-tight. The obtained results demonstrate a high at 1 ml min-1. Chromatograms were acquired at 370 sensitivity and reproducibility values expected for the nm excitation and 470 nm emission wavelength for technique, which allows to conclude that the method IRI, and emission at 534 nm for SN-38 and IS. The excludes many of the analytical errors present in method was validated according to international manual tests. guidelines. Results: The chromatographic run was of Financial support / Acknowledgments: Centro de 19 min. The method was linear from 10 to 3,000 ng Informação Toxicológica do Rio Grande do Sul mL-1 and from 1 to 300 ng mL-1 (r> 0.99) for IRI and SN-38, respectively, accurate, with CV% <15 intra- and inter-assay, and exact (85-115%). The extraction yield was around 42%. Analytes did not present instability. There were no interferences in the retention times of the analytes. Conclusion: The method ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 43 presented adequate characteristics for clinical use and on Hct 50%. The extraction yield was more than 42%. is being applied in an ongoing clinical study. Analytes did not present instability. There were no Financial support / Acknowledgments: Laboratório interferences in the retention times of the analytes. de Análises Toxicológicas Conclusion: The HPLC-FL method for determination of IRI and SN-38 in DBS presented suitable characteristics for the clinical use for therapeutic AN 13 - DETERMINATION OF IRINOTECAN monitoring of IRI. AND SN-38 IN DRIED BLOOD SPOTS Financial support / Acknowledgments: Laboratório EMPLOYING HIGH PERFORMANCE LIQUID de Análises Toxicológicas da Universidade Feevale. CHROMATOGRAPHIC COUPLED WITH FLUORESCENCE DETECTOR (HPLC-FL) AN 14 - EVALUATION OF PHTHALATE HAHN, R. Z. 1; COSTA, P. A. 1; CEZIMBRA, A. 1; MIGRATION IN BLOOD COMPONENTS BAGS ANDRIGUETTI, N. B. 1; SCHNEIDER, A. 1; REIS, BY GAS CROMATOGRAPHY (GC-FID/GC-MS) S. L. 2; KLÜCK, H. 2; SCHWARTSMANN, G. 2; ANTUNES, M. V. 1; LINDEN, R. 1 OLIVEIRA, W. M. L. 1; SANTOS, B. A. 1; ALVARENGA, R. B. 1,2; BRUM, D. E. L. 2; 1. Laboratório de Análises Toxicológicas, ROMÃO, P. R. T. 1; TSAO, M. 1; DALLEGRAVE, Universidade Feevale, Novo Hamburgo-RS, Brazil; 2. E. 1; MACEDO, S. M. D. 1 Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre-RS, Brazil. 1. Universidade Federal de Ciências da Saúde de Porto Alegre - RS, Brasil; 2. Irmandade Santa Casa de Background: Irinotecan (IRI) is an antineoplastic drug Misericórdia de Porto Alegre - RS, Brasil widely used for the treatment of colorectal and advanced pancreatic cancer. Despite its clinical utility, Introduction: Blood components are byproducts IRI and its active metabolite SN-38 present a narrow derived from blood through physical processes therapeutic window and its use is associated with (centrifugation and freezing) that are stored in plastic potentially severe hematopoietic and gastrointestinal bags made from PVC using phthalates as plasticizers. toxicities. The quantification of IRI and SN-38 in dried Di 2-ethylhexyl phthalate (DEHP) is the most blood spots (DBS) may be an alternative to commonly used plasticizer. The toxicity of DEHP is individualize the drug dose through a minimally related to fertility and embryo development, as invasive collection method. Objective: To develop and observed in male Winstar mice. It is known that high validate a method for the simultaneous determination dose intoxications cause a decrease in testicular weight of IRI and SN-38 in DBS by HPLC-FL. Method: and tubular atrophy in rodents. There is still little Whole blood containing IRI and SN-38 was applied to information on the amount of phthalates that is leached Whatman 903 paper and, after drying, an 8 mm to the blood components contained in plastic bags and diameter disc was extracted with 350 μL of the internal how much this factor may be related to transfusion standard solution (IS, camptothecin). Samples were incidents. Objective: This study aims to develop a incubated at 45 °C for 30 min under stirring. After, the methodology for extraction using QuEChERS to detect extractive phase was evaporated to dryness and phthalates in blood components, to standardize the recovered with 150 μL of a mixture of mobile reagents used in the extraction phase, and finally to phase:HCl 1 M. Separation was performed using an apply the method in samples of blood components. Eclipse Plus C8 (150 × 4.6 mm, 5 μm) column. The Material and Methods: Samples were obtained from mobile phase was a mixture of 0.1 M phosphate buffer the Hemotherapy Service at the Irmandade Santa Casa pH 4.0 and acetonitrile (80:20, v/v), at 1 mL min-1. de Misericórdia Hospital of Porto Alegre (ISCMPA), Chromatograms were acquired at the excitation extracted by the QuEChERS method, and analyzed by wavelength of 370 nm, with emission wavelengths of gas chromatography–mass spectrometry (GC-MS). 420 and 540 nm, for IRI and SN-38 plus IS, Blank samples were addiction with known respectively. The method was validated according to concentrations of the standard solution (DEHP). For international guidelines. Accuracy and extraction sample extraction, 500 ul were diluted in distilled efficiency were evaluated in different haematocrit. water and extracted with 2 mL of hexane. Sodium Results: The chromatographic run time was 16 min. acetate (100 mg) and magnesium sulfate (400 mg) The method was linear from 10 to 3,000 ng mL-1 and were added for the salting-out step, and for the from 1 to 300 ng mL-1 (r> 0.99) for IRI and SN-38, cleanup, PSA (25 mg), C18 (50 mg) and magnesium respectively, precise with intra- and inter-assay CV% sulfate (150 mg) were added to the extract. Results: So <15, and accurate (85-115%). Accuracy (92,5-108%) far, QuEChERS extraction and the chromatographic was slightly affected by the haematocrit, being the method have been standardized and validated for smaller value observed in the Hct 25% and the higher samples of blood components containing DEHP. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 44

Hexane is the solvent used in our method, providing a for screening and subsequent quantification of these clean extract with no interferents. The retention time psychoactive substances. Its use will focus on police for DEHP is 11,343 min and one sample run takes only blocks, post accident, violent behaviors and for 19 min. Conclusion: This methodology will be used analyzes that require immediate correlation. for analysis in the samples collected from the ISCMPA Financial support / Acknowledgments: Maxilabor to determine the concentration values of DEHP in Diagnósticos LTDA blood components.

AN 15 - DEVELOPMENT AND VALIDATION A KIT FOR DETECTION DRUGS OF ABUSE IN ORAL FLUID

GIATTI, M. A.1; ANDRIOLI, T. C.1; MORAES, A. A. T.1; MIRANDA, L. S.1; DE NARDI1, C. S.; WONG, A.1

1. Maxilabor Diagnósticos LTDA, Sao Paulo - SP, Brazil.

Introduction: The use of psychoactive substances (licit and illicit) must be investigated and requires a quick, precise response in specific scenarios as police blocks, accidents, airline pilots and workers assuming high risks situations. Drugs should be detected in saliva because it presents shorter window of detection (minutes to hours). Oral fluid is a non-invasive, easy to collect matrix with minimal potential for fraud and contamination that provides diagnosis of drug ingestion. We developed an innovative kit that allows screen detection of the drugs on site, confirmation test and split sample. Objective: To validate the screen results on the strips in the kit with an analytical method (LC-MS/MS) to identify and quantify cocaine, benzoylecgonine, anhydroegoninamethyl ester (AEME), amphetamine, methamphetamine, MDMA and MDA (ecstasy), codeine, 6-Acetylmorphine, morphine and tetrahydrocannabinol (THC) in saliva samples. Material and Methods: For screening we used our Presto Saliva TM kit developed by MaxiKit. The method was validated establishing the limit of detection (LOD) and quantification (LOQ), precision and accuracy, selectivity, robustness, carry over test and matrix effect. Saliva samples were extracted with methanol and analyzed by LC-MS/MS (Waters®). Results: In a linearity of the method the concentrations were 1, 2, 4, 8, 12, 15 and 20 ng/mL of analytes, such as cocaine, benzoylecgonine, AEME, amphetamine and methamphetamine, MDA, MDMA, THC, 6- acetylmorphine, Morphine and Codeine. Correlation coefficient (R) higher than 0.99. Stability evidence of at least 90 days with deviation less than 15% for all analytes. Confirmation of real samples that altered results in Presto Saliva showed a high sensibility to detection of THC in real samples (n=7). The concentrations were 0.32; 2.46; 17.91; 17.54; 24.11; 109.0 and 123.25 ng/mL for THC. Conclusion: This Presto SalivaTM kit is a reliable and sensitive method

CLINICAL TOXICOLOGY ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 46

CL 01 – CETIRIZINE - GABAPENTIN CL 02 - CLINICAL TOXICOLOGY AND INTERACTION IN PATIENTS WITH SAFETY STUDY OF HERBAL MEDICINES IN NEUROPATHIC PAIN: PHARMACOKINETICS VOLUNTEERS FROM A PHYTOTHERAPY CLINIC OF THE HEALTH PUBLIC SYSTEM COSTA, A.C.C. 1; YAMAMOTO, P.A. 2; BENZI, J.R.L. 1; LAURETTI, G.R. 3; DE MORAES, N.V. 2. CIRILO H.N.C (1,2); CARNEIRO D.M. (3); COSTA C.D.D. (4); CUNHA L.C. (5); BARA M.T.F. (1) 1. Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto - SP, (1) Programa de Pós-Graduação em Ciências Brasil; 2. Faculdade de Ciências Farmacêuticas, Farmacêuticas, Faculdade de Farmácia, Universidade Universidade Estadual Paulista, Araraquara - SP, Federal de Goiás, Goiânia-GO, Brasil. (2) Hospital das Brasil; 3. Faculdade de Medicina de Ribeirão Preto, Clínicas da UFG/ Empresa Brasileira de Serviços Universidade de São Paulo, Ribeirão Preto - SP, Brasil Hospitalares (EBSERH), Goiânia-GO, Brasil. (3) Centro de Especialidades em Práticas Integrativas e Introduction: Gabapentin (GBP), an anticonvulsant Complementares, Secretaria Estadual de Saúde de used in the treatment of neuropathic pain, is primarily Goiás, Goiânia-GO, Brasil. (4) Programa de Pós- eliminated by renal excretion, partially depending on Graduação em Ciências da Saúde, Faculdade de organic cation transporter 2 (OCT2) activity. The Medicina, Universidade Federal de Goiás, Goiânia- transporter, mainly expressed on the basolateral GO, Brasil. membrane of kidney proximal tubule, promotes the elimination of several clinically used drugs, by their Introduction: the use of herbal medicines and herbal uptake from blood into proximal tubule cells. products has been increasing worldwide, so as the Cetirizine (CET), an antiallergic used to treat rhinitis, reports of adverse events related to them. The World inhibits OCT2 activity both in vivo and in vitro. Health Organization Toxicity Grading Scale for Objective: The aim of this study was to determine Determining the Severity of Adverse Events (WHO- whether the OCT2 inhibitor CET influences GBP TGS) can be used in human clinical trials for grading pharmacokinetics in patients with neuropathic pain. abnormal laboratory parameters as evidences of Material and Methods: The Ethics Committee of the adverse events. Objective: to evaluate the safety and School of Pharmaceutical Sciences of Ribeirão Preto clinical toxicology of herbal medicines in healthy and the School of Medicine of Ribeirão Preto, volunteers, using the WHO Toxicity Grading Scale for University of São Paulo (USP) approved the clinical Determining the Severity of Adverse Events. Material protocol on April 16th, 2015 (CAAE: and methods: for this, an open, prospective clinical 34175314.3.0000.5403). Eight patients with trial was conducted in a complementary and neuropathic pain due to cervical/lombar disc integrative health unit, from State Department of herniations and pain ≥ 4 on visual analogue scale were Health of Goiás. The sample consisted of 35 investigated. Patients were treated with 300 mg GBP volunteers, recruited from October/2016 to April/2017, (Phase 1) and 20 mg/day of CET for five days and 300 considered healthy after laboratory tests which mg GBP on the last day of CET treatment (Phase 2), preceded the study. The laboratory tests included with a wash-out period of at least 15 days. Serial blood hematological, biochemical, serological and urine samples were collected up to 36 hours after GBP analysis. At the end of a 30-day period using the herbal administration. GBP plasma concentrations were medicines prescribed, the volunteers repeated the determined by HPLC-UV after derivatization using 1- laboratory tests and the results were analyzed using the fluoro-2,4-dinitrobenzene. Results: Pharmacokinetic WHO-TGS. Those who presented abnormal laboratory parameters were estimated by non-compartmental results, had the herbal medicine suspended for 30 days, analysis. The administration of CET decreased the and repeated the tests after that period. The results plasma concentrations of GBP (AUCPhase1 26950 ± were analyzed using the WHO-TGS. Results: 16 18250 ng.h/mL vs. AUCPhase2 21060 ± 15470 volunteers (45,7%) did not experience adverse events ng.h/mL, Wilcoxon test: p=0.0078) due to the higher after 30 days using the herbal medicines prescribed, total clearance (Cl/F) in Phase II (1.93 ± 1.0 L/h vs according to WHO-TGS. 19 volunteers (54,3%) 1.54 ± 0.96 L/h, p=0.0078). Conclusion: In conclusion, presented Grade 1 Toxicity adverse events, which the the interaction between CET-GBP might be clinically most common was the increase in amylase’s relevant because GBP AUC values were reduced when concentration. After 30 days without the herbal CET was coadministered. However, this interaction medicine, 8 subjects showed no adverse events, while was not mediated by renal OCT2 activity. 11 continued presenting Grade 1 Toxicity adverse Financial support / Acknowledgments: CNPq e events, the most common being the increase in Programa de Apoio ao Desenvolvimento Científico amylase’s concentration and hematuria, each one (PADC-FCF-UNESP). observed in 4 subjects. Grade 2 Toxicity hypomagnesemia was observed in 1 volunteer. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 47

Conclusion: this is an ongoing study, and these are paralysis, respiratory depression, and circulatory preliminary results. However, we could observe that failure. There is no antidote for TTX. Treatment monitoring patients in use of herbal medicines allows consists of supportive care. Early diagnosis is essential the identification of adverse events in a systematic and for better management and outcome. Educational objective way, favoring intervention processes. measures could be useful to prevent pufferfish poisoning. Financial support / Acknowledgments: Centro de CL 03 - DEATH AFTER PUFFERFISH INTAKE - Informações Toxicológicas de Santa Catarina– CASE REPORT AND TETRODOTOXIN CIT/SC/SUR/SES e HU/UFSC. REVIEW

DELGOBO, C. L. 1,2; TORRES, J. L. 1; WEBER, J. CL 04 - IS CERULOPLASMIN AS A COPPER F. 1; TAKAHASHI, C. Y. 1; RODRIGUES, C. K. 1; INDICATOR STILL VIABLE? ZANNIN, M. 1,3. PEREIRA, D. D.; PULCHINELLI, A. JR. 1; 1. Centro de Informações Toxicológicas de Santa VICENTE, G. R. Q. 1; FÉDOR, S. 1; FARSETTI, J. Catarina–CIT/SC/SUR/SES e HU/UFSC; 2. Depto de M.1. Análises Clínicas - ACL/CCS/UFSC - Florianópolis/SC; 3. Depto de Patologia e 1. Grupo Fleury, São Paulo - SP, Brasil. Toxicologia–PTL/CCS/UFSC - Florianópolis/SC. Copper is an essential metal present in various Introduction: Tetrodotoxin (TTX) is a non-protein, biological processes and bound to metalloproteins. thermo-stable neurotoxin first believed to be present Their intake ranges from 1 to 1.6 mg/day (minimum only in pufferfish of the family Tetraodontidae. Later, daily requirement of 0.9 mg) and the main source is it was found in a variety of organisms, including algae, cereals. Absorbed between 20 and 50% in the intestine arthropods, echinoderms, molluscs, worms and frogs. and transported bound to albumin. Stored in the liver TTX origin is unknown. In pufferfish, it seems to be (ceruloplasmin) and excreted in bile, feces and urine. It produced by bacteria, subsequently accumulating participates in several biological functions: energy through the food chain. Pufferfish consume is production; in connective tissue; Metabolism of Fe; commonly seen among fishing communities in Brazil CNS and anti-oxidant. The major diseases related to without poisoning, but TTX concentration vary copper metabolism: Wilson's disease, Fe-resistant between distinct species or different individuals of anemia; Menkes syndrome. Serum copper levels are same specie, making the risk unpredictable. Objective: greatly reduced in Menkes syndrome and normal or To report a case of death after puffer fish intake decreased in Wilson's disease. Both conditions are due attended by telephone by the Center for Toxicological to genetic defects involving the cellular incorporation Information (CIT) in Santa Catarina and review of copper and its hepatic excretion. Regardless of tetrodotoxin effects. Matherial and methods: Not metabolism or ingestion, some diseases increase serum applicable. Results: A 67-year-old female patient was levels of copper: pregnant women; Hodgkin's admitted on emergency service with reports of having lymphoma; rheumatoid arthritis; Biliary cirrhosis; started with malaise after pufferfish intake, presenting Thyrotoxicosis; Anemia, hemochromatosis, impaired level of consciousness and desaturation. She thalassemia; typhoid fever; Tuberculosis and suffered 3 cardiac arrests, with return after maneuvers myocardial infarction. The present study aims to of resuscitation and was referred to the reference evaluate ceruloplasmin as the indirect indicator of hospital, in mechanical ventilation without sedation, copper levels. A retrospective survey was performed maintaining Glasgow 3, in fixed mydriasis, without on the results of plasma copper and ceruloplasmin, deep reflexes, receiving vasoactive drug. About 8 analyzed in the laboratory routine of patients, hours after arrival at the hospital, medical staff performed between July 2016 and July 2017. Plasma contacted CIT for information on pufferfish poisoning. copper concentrations were quantified by atomic Patient's relatives reported history of previous heart absorption spectrophotometry and ceruloplasmin was disease. She maintained initial neurologic condition determined by nephelometry. The data were stratified and evolved with hemodynamic instability, going to by reference values and analyzed by statistical death about 14 hours after the ingestion. Discussion: program. 4,193 cases were collected and concomitant TTX acts by blocking voltage dependent sodium copper and ceruloplasmin analyzes were performed. receptors, preventing depolarization and propagation We found 655 (15.6%) altered cases of copper and 390 of action potential in nerve cells of peripheral motor, (9.3%) altered ceruloplasmin. Taking as a parameter sensory and autonomic nerves. TTX also acts by the levels of ceruloplasmin (high, normal and low), we depressing the respiratory and vasomotor center of the observed: normal ceruloplasmin does not detect 9.2% encephalic trunk. Death occurs due to muscle of altered copper (high and low); ceruloplasmin high in ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 48

1.7% of cases erroneously classified normal copper as FITOPROT, three times daily, for ten consecutive high; and in 38% of cases low ceruloplasmin classified days. No participant experienced toxicity or normal as low. We conclude that, despite its low cost unacceptable discomfort and/or adverse reactions and analytical ease, ceruloplasmin does not perform (CTCAE v5.0), with laboratory and clinical parameters compatible with plasma copper dosage. under normal conditions. Side effects observed were Financial support / Acknowledgments: Grupo low intensity and temporary mucosa/dental surface Fleury. pigmentation (n=7) and tooth sensitivity (n=4), which disappeared after formulation use ceased. No significant cellular genotoxic effects were observed CL 05 - EFFECTS OF A MUCOADHESIVE (P>0.05). Biochemical assays reveled no altered levels FORMULATION CONTAINING of myeloperoxidase (P=0.2268), malondialdehyde CURCUMINOIDS (ZINGIBERACEAE) AND (P=0.1188) nor nitric oxide (P=0.5709) concentration, BIDENS PILOSA L. (ASTERACEAE) EXTRACT and no significant difference were found in the levels FOR PREVENTION AND TREATMENT OF of pro-inflammatory cytokines (P>0.05). As ORAL MUCOSITIS: in vitro AND CLINICAL conclusion, FITOPROT protected HaCaT cells against PHASE I STUDY 5-FU-induced damage exerting chemoprotective activity; and FITOPROT demonstrated to be safe and SANTOS FILHO, E. X. 1; SILVA, A. C. G. 1; tolerable, been suitable for evaluation in a phase II trial ÁVILA, R. I. 1; ARANTES, D. A. C. 2; BATISTA, A. as treatment against OM. C. 2; MENDONÇA, E. F. 2; MARRETO, R. N. 3; Financial support / Acknowledgments: Conselho LIMA, E. M. 4; VALADARES, M. C. 1. Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq); Coordenação de 1 Laboratory of Pharmacology and Cellular Aperfeiçoamento de Pessoal de nível Superior Toxicology, Faculty of Pharmacy, Federal University (CAPES); Financiadora de Estudos e Projetos of Goiás, Goiânia - GO, Brazil; 2 Laboratory of Oral (FINEP). Pathology, Dental School, Federal University of Goiás, Goiânia - GO, Brazil; 3 Laboratory of Nanosystems and Modified-Release Drugs Devices, Faculty of CL 06 - ACUTE RENAL FAILURE AFTER Pharmacy, Federal University of Goiás, Goiânia - GO, BOTHROPIC ACCIDENT: CASE REPORT Brazil; 4 Laboratory of Pharmaceutical Technology, Faculty of Pharmacy, Federal University of Goiás, RODRIGUES, E.C.A.M.S.1,3; COUTINHO, M.S.1,3; Goiânia - GO, Brazil FOOK, S.M.L.1,3; SOUZA, D.R.1; SILVA, R.C.1; CAVALCANTE, J. S.1; MARIZ, S. R.2,3 Toxicity related to anticancer therapies is a huge public health problem to be overcome. Mucositis is a serious 1 Universidade Estadual da Paraíba, Campina Grande - limiting side effect observed in patients under PB, Brasil; 2 Universidade Federal de Campina antineoplastic therapies and still no approach was Grande, Campina Grande - PB, Brasil; 3 Centro de completely successful in treating it. Preclinical Informação e Assistência Toxicológica de Campina repeated-dose toxicity and efficacy studies developed Grande, Campina Grande - PB, Brasil. by our group have suggested the potential of a mucoadhesive formulation containing curcuminoids Introduction: Ophidic accidents are an important and Bidens pilosa L. extract (FITOPROT) in treating public health problem due to their high incidence and mucositis. Here, we evaluated the effects of severity. In Brazil, snakes of the Bothrops genus are FITOPROT as a protective agent against 5-FU-induced responsible for the majority of accidents, and one of cellular toxicity in vitro; and establish the safety of the most serious complications is acute renal failure FITOPROT for the prevention and treatment of (ARF), present in 2-10% of cases. Objective: Report chemoradiotherapy-induced oral mucositis (OM). In the case of a patient attended by the Toxicological the in vitro study, HaCaT cells were pretreated with Assistance Center of Campina Grande-PB (CEATOX- 0.005% of FITOPROT for 24 hours, and then treated CG), who presented ARF after bothropic accident. concomitantly with FITOPROT and 5-FU (10 µg/mL) Materials and Methods: S.C.S., 36 years old, woman, for more 24 hours. FITOPROT significantly reduced farmer resident of Sumé-PB, arrived at the hospital in ROS generation, reversed mitochondrial membrane Campina Grande-PB six hours after bothropic accident depolarization and cellular proliferation loss; on right foot, reporting local pain and vomiting modulated oxidant response, apoptosis and episodes. The exams presented complete blood inflammation by reducing respectively Nrf2, TNF, incoagulability. Three ampoules of antibothropic cytochrome c, NF-κB and pro-inflammatory cytokines serum (ABS) was administered initially to the patient. (IL-1β, IL-6, IL-8) levels. In the phase I study, twenty 24 hours after ABS, the exams presented a recovery of healthy adult participants rinsed their mouths with coagulation, but showed leukocytosis (15.100 mm3), ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 49 thrombocytopenia (65.000 mm3) and rapid worsening University of Paraíba (Protocol of renal function (urea 106 mg/dL; creatinine 6,0 57562816.4.1001.5187). Results: In this study, 536 mg/dL). When activated, CEATOX-CG directed the active ingredients were observed, most of them being administration of additional serum therapy (totaling 10 insecticides and rodenticides. Most of the poisoning ampoules of ASB), venous hydration and diuretics. occurred by the chemical group Carbamates (33.9%), However, the patient developed oliguria, dyspnea and whose main pesticide used was the "chumbinho" anasarca, requiring hemodialysis. After 30 days of (28.7%). Using the variable "suicide" as a dependent, hospitalization, and with improvement of the general we have a statistically significant association at the condition, the patient was discharged from hospital, junction with the variables "sex", "age", "labor market but with renal function not reestablished and in a situation" and "case evolution". In the Binary Logistic regular dialysis program. Results: It is known that after Regression model, we observed an association with two hours of the bothropic accident, glomerular "Gender" (p = 0.032) and "Age" (p = 0.001). deposition of fibrin begins and acute tubular necrosis Outbreaks were identified in three districts of the (ATN) can already be observed. The proteolytic, municipality: Malvinas (West Zone) with 23 records, hemorrhagic, coagulant and nephrotoxic action of Catolé (South Zone) with 18 records and José Pinheiro bothropic venom leads to vascular occlusion and renal (East Zone) with 11 records. For the calculation of the ischemia, and when associated with hemodynamic Moran Index, we obtained IMo = -0.006725361 (p = disorders and immunological reactions, culminate with 0.675), and there was no statistical evidence to prove the appearance of ARF. Conclusion: The favorable the spatial correlation between the neighborhoods of prognosis of ARF after botropic accident is related, Campina Grande and the continuity of the occurrence among other factors, to the early implementation of the of pesticide poisonings. Conclusion: The easy access specific treatment and, when necessary, hemodialysis to pesticides due to unregistered and illegal sales and should be initiated soon after the onset of renal popular markets, contrary to what is provided for in changes. Law 7802/1989, favor the increase of intoxications by Financial support / Acknowledgments: CAPES. these substances. Financial support / Acknowledgments: CAPES.

CL 07 - TOXICOLOGICAL INCIDENTS RELATED TO PESTICIDES: A SPATIAL AND CL 08 - CYPA4*1B, CYP3A5*3, GSTT1 AND EPIDEMIOLOGICAL ANALYSIS GSTM1 POLYMORPHIMS NOT INFLUENCE ON CHRONIC MYELOID LEUKEMIA RODRIGUES, E.C.A.M.S.1,3; COUTINHO, M.S.1,3; TREATMENT RESPONSE FOOK, S.M.L.1,3; SOUZA, D.R.1; MARIZ, S. R.2,3. SILVEIRA-LACERDA, E.P. 1; DELMOND, K. A. 1; 1 Universidade Estadual da Paraíba, Campina Grande - ABREU, D. C. 1; PINTO, T. M. 1; GOVEIA, R.M. 1. PB, Brasil; 2 Universidade Federal de Campina Grande, Campina Grande - PB, Brasil; 3 Centro de 1. Universidade Federal de Goiás, Goiânia – GO, Informação e Assistência Toxicológica de Campina Brasil Grande, Campina Grande - PB, Brasil. The Imatinib Mesylate (IM) - Gleevec™ act as Introduction: The possibility of exposure and selective inhibitor of BCR–ABL tyrosine kinase contamination by artificial and natural toxic substances activity on Chronic Myeloid Leukemia (CML) is high, when considering the various forms of treatment. Despite this cases of treatment failure or of contamination and the various toxic agents, such as suboptimal response are sometimes seen. CYP3A4 and pesticides, responsible for intoxications with greater CYP3A5 are polymorphic genes that encoding clinical severity. Objective: Investigate the profile of metabolic enzymes involved in the oxidative pesticide intoxications registered in Toxicology metabolism of IM and Glutathione S-transferases Assistance Center of Campina Grande, Paraíba, Brazil (GSTs) enzymes act in the metabolism of xenobiotics, (CEATOX-CG). Materials and Methods: This is a including chemotherapeutic drugs. This study was quantitative, ecological, nested study of a cross- carried out with CML patients Goias (Brazil) residents sectional epidemiological. Data were collected through in order to associate the genotype of patients for Sinan's Toxicological Events Notification Form polymorphims in metabolic genes with cytogenetic and occurring in the municipality of Campina Grande, molecular responses during treatment with IM. The between 2011 and 2015. SPSS software v. 20.0 was study was conducted in accordance with the used for statistical analysis. For the georeferencing, Declaration of Helsinki and was approved by ethics QGIS software, v. 2.14.3. The Moran Index was committee (Comitê de Ética em Pesquisa do Hospital calculated through R Core Team. This study was das Clínicas da Universidade Federal de Goiás). A approved by the Ethics Committee of the State total of 114 CML patients in use of IM from Hospital ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 50 das Clínicas-UFG, Goiânia, Goiás, Brazil, were abdominal pain. The diagnosis was made through enrolled in this study. Response criteria were based on clinical manifestations, laboratory tests and European Leukemia Net recommendations. identification of the caterpillar by photos taken by the Genotyping was carried out for CYP3A4*1B (- patient. By the means of Clotting Time (CT) test, 392A>G (rs2740574)), CYP3A5*3 (6986A>G Prothrombin Time (PT), Activated Partial (rs776746)), GSTT1*0 (null allele) and GSTM1*0 Thromboplastin Time (aPTT), Prothrombin Activity (null allele) polymorphisms by q-PCR. Statistical (PT% activity), Complete Blood Count (CBC), urea analysis was carried out using GraphPad Prism5® and creatinine levels, blood incoagulability was software. The X2-test or Fisher’s exact test was used to detected. Coagulopathy is one of the main laboratory compare response and specific genotypes (α=0,05). manifestations of Lonomia sp. poisoning. The The results revealed no statistical difference between consumption of coagulation factors and the activation evaluated polymorphisms CYP3A4*1B, CYP3A5*3, of secondary fibrinolysis causes haemorrhage, due to GSTT1*0 and GSTM1*0 and responses to treatment the presence of two procoagulant toxins in Lonomia with IM (P>0,05). The same result was seen when sp. bristles: Losac, a factor X activator, and Lopap, a comparison was performed with polymorphisms prothrombin activator. The patient received five grouped for each patient (P>0,05). These findings have ampoules of antilonomic serum therapy (ALS), shown that in the studied population the normalizing her CT, PT and aPTT levels, and she was polymorphisms CYP3A4*1B, CYP3A5*3, GSTT1*0 discharged 37 hours after her admission. The use of and GSTM1*0 do not influence the performance ALS therapy was crucial for normalizing her blood efficiency of the drug (IM). coagulability and avoid further complications. This Financial support / Acknowledgments: FAPEG; type of events is recognized as a health problem, but CAPES. health actions are still needed to increase the prevention of venomous animals’ accidents. Financial support / Acknowledgments: Universidade CL 09 - COAGULATION ABNORMALITIES Estadual da Paraíba (UEPB)/ Centro de Assistência ASSOCIATED WITH LONOMIA SP. IN THE Toxicológica de Campina Grande (Ceatox-CG). NORTHEAST OF BRAZIL: A CASE REPORT

SILVA, R.C. 1; SILVINO, M.R.S. 1; CORDEIRO, CL 10 - EPIDEMIOLOGICAL PROFILE OF G.B.C. 1; ASSIS, L.R. 1; VASCONCELOS, M.E.D. 1; DRUG POISONINGS REGISTERED BY THE CAVALCANTE, J.S. 1; COUTINHO, M.S. 1,2; POISON CONTROL CENTRE OF CAMPINA RODRIGUES, E.C.M.S. 1,2; FOOK, S.M.L. 1,2. GRANDE (CEATOX-CG) BETWEEN 2009 AND 2015 1. Universidade Estadual da Paraíba, Campina Grande- PB, Brasil; 2. Centro de Assistência Toxicológica de CORDEIRO, G.B.C. 1; MACHADO, V.N. 1; Campina Grande (CEATOX-CG), Campina Grande- DANTAS, T.L. 1; VASCONCELOS, M.E.D. 1; PB, Brasil. BRAGAGNOLI, G. 2; MARIZ, S.R. 2.

The genus Lonomia is one of the most important 1. Universidade Estadual da Paraíba - Campina within the order Lepidoptera, given its clinical Grande-PB, Brasil; 2. Universidade Federal de relevance in Brazil. This caterpillar is in the larval Campina Grande - Campina Grande-PB, Brasil. form of the Lepidoptera life cycle, and causes damage in agriculture and serious accidents in humans, According to recent data from the National Poisoning presenting a clinical picture characterized by Information System (SINITOX), drug poisonings in coagulation abnormalities, bleeding gums, urticaria, Brazil lead the statistics, which mainly affects females and acute renal failure. These accidents are caused by aged between 1 and 4 years old. We traced a small poison inoculum structures called scolus, present transversal, retrospective, descriptive and analytic in the caterpillar bristles, small spines filled by toxic epidemiological profile of acute drug poisonings secretion. This is a case report of an accident caused treated in a hospital unit between 2009 and 2015, and by Lonomia sp. treated in a hospital unit and registered registered by Ceatox-CG on Information System on by the Poison Control Centre of Campina Grande Diseases of Compulsory Declaration (SINAN) sheets. (Ceatox-CG). A 35-year-old female, teacher, resident We collected data from SINAN files, processed in in the urban zone accidentally stepped on the Microsoft Excel®, and analysed them using SPSS caterpillar, coming to skin contact with the bristles Statistics Base 17.0. The variables were: seasonality, through the plantar aspect of the foot. She was the area of occurrence, circumstance, sex, age, hospitalised 3 hours after the event and reported occupation, and toxic agent (groups of drugs). There intense burning pain, local oedema and erythema in the were 1594 cases of drug poisonings, predominantly plantar region of the foot, headache, nausea, and with a female profile (65,62%), student (28,86%), ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 51 from 20 to 29 (22,71%) and 1 to 4 (16,75%) years old, 2015, data were collected in the National Sanitary on a suicidal attempt (54,27%) or individual accident Surveillance System - Goiás (NSSS), with the (19,95%). These results are similar to Morais et al, objective to establish the profile of the individuals with 2008¹, except for the increment on poisonings in intentional intoxication per aldicarb. Where the children from 1 to 4 years old, which modified the variables were extrated, the frequency of events by epidemiological profile in Campina Grande, making it toxic agents and the number of suicide attempts. more like the national profile registered by SINITOX. Results: Of the 1237 fiches registered in the NSSS of Within the occurrences caused by only one agent intoxication by aldicarb, 1046 are suicide attempts. (82,37%), neuropsychological drugs were present in The highest occurrences were in the years of 2010 61,61%, and psycholeptics drugs predominated (22.84%) and 2012 (20.45%). Conclusion: It is (63,91%). The association between neuropsychological concluded that the population has easy access to drugs and sex was statistically significant (p = 0.016), aldicarb in the veterinary and agricultural products indicating a greater female participation in the use of trade, even though it is considered clandestine. It this class of drugs. Likewise, there was a significant becoming a great challenge to make the population correlation (p=0.000) between sex and circumstance, aware of the high toxicity of the product. showing that women are more likely to use drugs in Financial support / Acknowledgments: Secretaria suicide attempts. The profile of poisoned patients Estadual de Saúde, vigilância sanitária/CIT. registered in Campina Grande between 2009 and 2015 was predominantly female, aged between 20 and 29 years old, and in use of neuropsychiatric medicines on CL 12 - EPIDEMIOLOGICAL PROFILE OF a suicide attempt. There was also an increase on drug POISONING BY TOXIC PLANTS IN GOIÁS poisonings in children aged 1-4 years, therefore FROM 2011 TO 2015 important for readjusting public health actions and knowledge. SILVA, L. A. 1; NASCIMENTO, H. A. 2; PRADOS, Financial support / Acknowledgments: Universidade V. O. 3; MACHADO, H. C. P. 4; OLIVEIRA, R. R. 5; Estadual da Paraíba/ Centro de Assistência MORAIS, L. T. 6; MAYA, A. C. L. 7; REZENDE, G. Toxicológica de Campina Grande. M. S. 8.

1 Centro Universitário de Anápolis, Anápolis - Goiás, CL 11 - THE INADEQUATE USE OF ALDICARB Brasil; 2. Centro Universitário de Anápolis, Anápolis - IN THE SELF-EXTERMINATION TRY IN THE Goiás, Brasil; 3. Centro Universitário de Anápolis, STATE OF GOIÁS, 2010 – 2015 Anápolis - Goiás, Brasil; 4. Centro Universitário de Anápolis, Anápolis - Goiás, Brasil; 5. Centro OLIVEIRA, K. C. J.; OLIVEIRA, K.C.C. 1; ROCHA, Universitário de Anápolis, Anápolis - Goiás, Brasil; 6. P.F. 2; SELLEGUIM, S.M.C. 3. Centro Universitário de Anápolis, Anápolis - Goiás, Brasil; 7. Centro Universitário de Anápolis, Anápolis - 1 Centro de Informação Toxicológica, Vigilância Goiás, Brasil; 8. Centro Universitário de Anápolis, sanitária, Goiania-GO, Brasil. Anápolis - Goiás, Brasil.

Introduction: Exogenous intoxications are a constant, INTRODUCTION: Exogenous intoxication can be constituting a relevant quantitative of cases, especially defined as the clinical and/or biochemical consequence those resulting from the ingestion of carbamate, also of exposure to chemicals found in the environment or known as "aldicarb". It is noteworthy that this isolated. OBJECTIVE: To discuss quantitatively and substance, although mistakenly known as raticide, is qualitatively the poisoning by toxic plants in Goiás in sold illegally, whose use has been popularized in large the period from 2011 to 2015. METHODOLOGY: urban centers. The processes of human intoxication This is a descriptive epidemiological study, whose data have constituted one of the most serious problems of were obtained by consulting the SINAN (Notification Public Health due to the lack of strategies of control of Injury Information System) database available by and prevention of intoxications. Aldicarb intoxication the Department of Informatics of the Unified Health is considered an important cause of morbidity and System (DATASUS).We used the free Geographic mortality in Brazil due to its easy access, low cost and Information System (GIS) QGIS and performed effectiveness front of suicide attempts. Objective: To geoprocessing techniques to visualize, edit, create and emphasize the importance of a greater awareness of the analyze the reference map data. The base and thematic society regarding the high toxicity of the aldicarb to maps were elaborated with the symbologies of the the human being, and to highlight the epidemiological cartographic normalization of the Technical Manual of characteristics of the cases of intoxication by aldicarb. Cartographic Conventions of the Brazilian Army. Material and Method: This is a descriptive and RESULTS: Considering exogenous intoxication, toxic quantitative study, relative to the period from 2010 to plants poisoning was 9.40% prevelant. The counties ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 52 that presented the most notifications were Goiânia, received 7,950 cases, of which 235 were reports of Formosa, Anápolis, Rio Verde, Valparaiso de Goiás therapeutic errors (2.96%). Children younger than 5- and Jataí. There was a higher prevalence in the female years-old were involved in 38.7% of cases and 22.1% gender (53.23%). Schooling was not related to were adults. Females accounted for 52.8% of the total incidence. The highest rates are in the age groups of 1- cases. Most of the therapeutic errors occurred at home 4 years (28.05%) and 20-39 years (38.55%), most of (91.5%) and 7.7% in a healthcare setting. The main them being accidental. There were more cases in the route of exposure was ingestion (87.7%). The most urban area (91.83%). Regarding the type of exposure, common therapeutic errors circumstances were: wrong 87.46% were classified as acute-single, 3.13% as medication given (45.1%), incorrect dose (17.8%), acute-repeated and 0.25% as chronic. Also, 86.64% inadvertently took/given medication twice (10.6%) and evolved to cure without permanent damages, 2.08% to iatrogenic error (8.5%). The main groups of cure with permanent damages and 0.71% to death due medications involved were those acting at the nervous to intoxication. CONCLUSION: The results are system (29.7%), respiratory system (11.8%) and justified by the lack of knowledge of the toxicity of sensory organs (10.6%). The risk of children ≤ 5 years some plants and the greater contact of women with old to experience a therapeutic error with respiratory them. In addition, the population believes that using drugs was 2.3 times superior to the risk of plants, including for treatment of diseases, is natural, experiencing an event involving other type of agent safe, cheap and effective. It is essential to point out (OR:2.34, CI95%:1.75-3.13, p<0.00001). Conclusion: that the possible relation of the greater prevalence of Most therapeutic errors involved children ≤ 5 years cases in the urban zone with the underreporting old, there is a predominance of the female gender and hampers exact data for research. the main route of exposure was ingestion. Drugs most Financial support / Acknowledgments: Conselho involved were those acting at the nervous system level. Nacional de Desenvolvimento Científico e Therapeutic errors are frequent preventable events at Tecnológico (CNPq). global level and Chile is part of that reality. It is important to reinforce strategies to reduce the impact of this type of events in order to achieve an effective CL 13 - CHARACTERIZATION OF approach for the prevention of poisonings. CONSULTATIONS RELATED TO THERAPEUTIC ERRORS RECEIVED AT THE TOXICOLOGICAL INFORMATION CENTER CL 14 - CHARACTERISATION OF CALLS OF THE PONTIFICIA UNIVERSIDAD FROM ELDERLY INDIVIDUALS TO THE CATÓLICA DE CHILE (CITUC) DURING THE TOXICOLOGICAL INFORMATION CENTER PERIOD BETWEEN JANUARY AND MARCH, PONTIFICIA UNIVERSIDAD CATÓLICA DE 2016 CHILE, PERIOD 2006-2016

CID-FLORES, L. 1; SILVA, L. 1; CERDA, P. 1; SILVA, L. A. S.; SILVA L, CERDA P, OLIVARES P, OLIVARES, P. 1; ROJAS, M.F. 1; RÍOS, J.C. 1,2. CID L, ITURRA P, MEDEL P, RÍOS JC.

1 Centro de Información Toxicológica de la Pontificia 1 Centro de Información toxicológica, Facultad de Universidad Católica de Chile, Santiago, Chile. 2 Medicina, Pontificia Universidad Católica de Chile, Departamento de Laboratorios Clínicos, Escuela de Santiago, Chile; 2 Department of Clinical Laboratory, Medicina, Pontificia Universidad Católica de Chile, Facultad de Medicina, Pontificia Universidad Católica Santiago, Chile. de Chile, Santiago Chile.

Introduction: Therapeutic errors are frequent events In Chile, the National Socioeconomic Characterization and have been widely studied in several countries, but Survey of 2015 revealed that individuals over 60 years in Chile there is no comprehensible information old correspond to 17.5% of the national population, regarding the subject. Objective: To characterize with a marked increasing tendency over the years. Due consultations related to therapeutic errors received at to this fact, it is necessary to analyze the profile of CITUC during the period between January 1st and exposures to chemicals in this age group in order to March 31st of 2016. Methods: A descriptive and adopting management measures that contribute to retrospective study of therapeutic error consultations improve their quality of life. The objective of this registered in CITUC between January and March, study was to characterized calls from individuals over 2016, was performed. Reports were recovered from the 60 years old received by the Poison Control Center of IPCS Intox DMS database. For the classification of the Pontificia Universidad Católica de Chile (CITUC) drugs the Anatomical Therapeutic Chemical during 2006 to 2016. A retrospective descriptive study Classification System developed by the WHO was was carried out considering calls received by the used. Results: Within the studied period CITUC Center between 2006 and 2016 that were ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 53 systematically stored using INTOX Data Management DBS and venous blood from 30 patients receiving FLU System. The following variables were analyzed: Age, were obtained. The concentrations of FLU and NFLU sex, agent, interlocutor, place of the incident, in DBS and plasma were determined by LC-MS/MS. circumstance of exposure and origin of the call. 11,744 From the determinations in DBS, plasma calls were received in this period, 64% of them concentrations (PC) were estimated in three different involving females. The mean age of individuals was ways: from an equation considering the individual 70.5 years. 41% of the calls were accidental exposures, Hct% of the patient, by multiplying by an equivalence followed by a 17% of therapeutic errors and 14 % of factor (0.71 for FLU and 0.68 for NFLU) and by the intentional suicidal exposures. The main agents Passin-Bablok regression equation, in order to evaluate involved were pharmaceutical products (48%), the best alternative. Results: Plasma quantifications cleaners/detergents (12%) and animals/arachnids- ranged from 18.2-517.9 for FLU and from 22.8-328 ng related calls (9%). Pharmaceutical products most mL-1 for NFLU, with [FLU]/[NFLU] ratio ranging commonly involved corresponded to neurological from 0.14 to 2.75. Concentrations obtained in DBS agents (53%), followed by cardiovascular agents showed high correlation with plasma quantifications (12%) and gastrointestinal and metabolic agents (7%). (r=0.982 for FLU and r=0.965 for NFLU, p<0.001). Within the neurological category, anxiolytics The estimated plasma levels were from 14.1-416.9 for represented 26%, antidepressants 17% and FLU and from 25.1-327.8 ng mL-1 for NFLU, with antipsychotics 15%. In all intentional exposures, values ranging from 0.12 to 3.10 for [FLU]/[NFLU] multiple agents were involved. Females exhibited a ratio. The use of an equivalence factor was considered higher risk of suicidal attempts with pharmaceutical more advantageous since it presented better products compared to males and other agents (OR 4.9; performance and excludes the need to apply an IC95% 3.3-7.4; p<0.0001). In this study, equation. Bland and Altman plots showed only two pharmaceutical products were the most common agent estimated concentrations estimated the range of ±1.96 involved. Due to their physiological conditions, this standard deviations, characterizing the comparability group of age presents particular conditions that may of the methods. Conclusion: These findings suggest affect metabolism and excretion and likewise the that DBS quantifications are representative of potential effects of pharmaceutical products. measurements obtained in plasma, indicating a Therefore, it is necessary to implement programs probable applicability of this sampling strategy in the aiming to improve the quality of life and reducing FLU TDM. sanitary costs. Financial support / Acknowledgments: Universidade Feevale; Capes

CL 15 - THERAPEUTIC DRUG MONITORING BY DRIED BLOOD SPOT: AN ALTERNATIVE CL 16 - ULTRA-HIGH PERFORMANCE LIQUID FOR FLUOXETINE THERAPEUTIC CHROMATOGRAPHY TANDEM MASS MONITORING SPECTROMETRIC METHOD FOR THE DETERMINATION AND VALIDATION OF SILVA, A. C. C. 1; PETEFFI, G. P. 1; BASTIANI, M. FLUOXETINE AND NORFLUOXETIN IN DRY F. 1; RAASCH, J. R. 1; ANTUNES, M. V. 1; HAHN, BLOOD IN DRIED BLOOD SPOTS R. Z. 1; PERASSOLO, M. S. 1; LINDEN, R. 1. SILVA, A. C. C. 1; PETEFFI, G. P. 1; BASTIANI, M. 1 Universidade Feevale, Novo Hamburgo - RS, Brasil. F. 1; RAASCH, J. R. 1; ANTUNES, M. V. 1; HAHN, R. Z. 1; PERASSOLO, M. S. 1; LINDEN, R. 1. Introduction: Fluoxetine (FLU) has become the most prescribed antidepressant, being present in several 1. Universidade Feevale, Novo Hamburgo - RS, Brasil. clinical and forensic cases. FLU is metabolized by CYP2D6 to the active metabolite norfluoxetine Introduction: Fluoxetine (FLU) is an antidepressant (NFLU), with a therapeutic window of 120-500 ng belonging to the class of selective serotonin reuptake mL-1 for FLU+NFLU, associated with [FLU]/[NFLU] inhibitors. It is metabolised by CYP2D6 to the active ratio of 0.7-1.9. Approximately 30-40% of patients metabolite norfluoxetine (NFLU), with a therapeutic treated with FLU do not present adequate response, window of 120 to 500 ng mL-1 for the sum of FLU + mainly due to interindividual pharmacokinetic NFLU. Approximately 30 to 40% of patients treated variations, making FLU a drug of interest for with FLU do not respond adequately, mainly due to therapeutic drug monitoring (TDM). Dry blood spots interindividual pharmacokinetic variation, making (DBS) obtained by capillary puncture can be used as FLU a drug of interest for therapeutic monitoring an alternative. Objective: To evaluate the applicability (TDM). Considering the difficulties of venous of the DBS in the TDM of FLU, comparing with collection, dry blood spots on paper (DBS) obtained by plasma determinations. Methods: Paired samples of capillary puncture can be used. Objective: To develop ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 54 and validate a method for determination of FLU and of anticholinergic syndrome from Bothrops antivenom NFLU in DBS by LC-MS/MS. Methods: DBS discs (BAV). Purpose: Describe a case report of a ophidic were obtained by fingerpricks and applied in Whatman accident caused by a bothropic serpent, evolving to a 903 paper. DBS were punched and an 8 mm disck was anticholinergic syndrom. Method:The data was used for extraction. Extration solveent was a mixture collected and analyzed from the notification cards used of methanol:acetonitrile (3:1, v/v) containing the at Ceatox unit in Campina Grande.Results: A 71 year- internal standard (IS) fluoxetine-D6. Separation was old female was hospitalised 3 hours after an ophidic performed in an Accucore C18 column (100x 2.1 mm, accident affecting the 2nd right toe. She was in a good p.d. 2.6 μm) eluted at a mobile phase flow rate of 0.4 general condition; her blood pressure was 170/100 mL min-1. Mobile phase was composed of formic acid mmHg, her heart rate was 85 beats/minute, with no 0.1% (A) and acetonitrile plus 0.1% formic acid (B). history of Systemic Arterial Hypertension. She Precision and accuracy were evaluated at reported local pain, with visible but discrete local concentrations of 40, 200 and 600 ng mL-1. The edema and bruising. According to coagulation acceptance criteria for accuracy were mean values standards her blood was incoagulable, which within ± 15% of the theoretical value and for precision confirmed that the accident was caused by a snake a maximum CV of 15%. Results: Analytical run time from the bothrops genre. However, it was a mild case was 11 min, with retention times for FLU and IS of 4.8 due to the symptoms present. The patient first received min and 4.6 min for NFLU. Intra-assay imprecision oral captopril 25 mg, and 3 ampoules of BAV were in the range of 4.68-7.83% and 3.54-7.99%, preceded by promethazine and hydrocortisone. Her while inter-assay imprecision were 3.13-9.61% and bruise first got worse, and she had episodes of delirium 4.77-7.7%, for FLU and NFLU, respectively. and agitation in the course of treatment, which was Accuracy were estimated at 97.9-110.4% for FLU and being given according to the protocol for BAV use. 100.2-104.08% for NFLU. The determinations in DBS Due to suspected hemorrhagic stroke, a Computerised showed a high correlation with plasma quantifications Tomography scan was conducted, but nothing with r=0.982 for FLU and r=0.965 for NFLU. significant was detected. Five days later, her symptoms Conclusion: The first method for determination of and coagulation tests normalised. On the 6th day, the FLU and NFLU in DBS using LC-MS/MS was patient showed significant improvement in her mental developed and validated. The assya fullfills the status, and was discharged on the following validation criteria for bioanalytical methods. DBS day.Conclusion: According to the patient’s quantification of FLU and NFLU showed high pharmacokinetic profile, a therapeutic dosage of correlation with plasma determinations, allowing its promethazine may cause adverse effects. Therefore, it use in clinical practice. was concluded that the use of promethazine as Financial support / Acknowledgments: Universidade premedication has caused this syndrome, which Feevale; Capes. normalized as the treatment ended. Financial support / Acknowledgments: Programa de Bolsa de Extensão da Universidade Estadual da CL 17 - ANTICHOLINERGIC SYNDROME DUE Paraíba (PROBEX/UEPB). TO PROMETHAZINE USED AS PREMEDICATION IN RESPONSE TO A BOTHROPIC ACCIDENT: A CASE REPORT CL 18 - EXPOSURE TO CIGARETTE SMOKE TOXICANTS IN SMOKERS WHO SWITCH TO BRAGA, M.L.F.M. 1; BARROS, A.M.G. 2; USING THE GLOTM TOBACCO HEATING CORDEIRO, G.B.C. 3; SILVA, L.M.R. 4; PRODUCT FERNANDES, W.O. 5; FOOK, S.M.L. 6. Gale, N. 1; Fearon, I.M. 1; McEwan, M. 1; Murphy, J. 1. Faculdade de Ciências Médicas de Campina Grande 1; Proctor, C. 1; Saraiva, A. 1. (FACISA); 1,2,3,4,5,6. Centro de Informação e Assistência Toxicológica de Campina Grande 1. British American Tobacco, R&D Centre, (CEATOX-CG); 2,3,4,5,6. Universidade Estadual da Southampton SO15 8TL, United Kingdom. Paraíba (UEPB). Intro: Tobacco heating products (THPs) represent a Background: Promethazine is a H1 receptor-blocking subset of the next-generation nicotine and tobacco agent, which is used in the premedication for the product category, in which tobacco is typically heated administration of antivenom in accidents caused by at temperatures of 250-350°C instead of burning venomous animals in Campina Grande, Brazil. The use (900°C) and has the potential to significantly reduce of promethazine might cause symptoms of the majority of cigarette smoke toxicants. The use of anticholinergic syndrome at high dosages. However, THPs holds great potential for reducing the harm there is no evidence in the literature of the occurrence associated with tobacco use but this needs to be ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 55 scientifically proven. Obj.:To examine changes in enzyme. The identification of patients with reduced biomarkers of exposure (BoE) to cigarette smoke DPD activity is mostly based on the measurement of toxicants when smokers switch to using the gloTM the concentrations of the endogenous compound uracil THP for 5 days, compared with those seen in smokers (U) and its DPD metabolic product, 5,6-dihydrouracil who remain smoking combustible tobacco cigarettes. (UH2) in plasma samples. Recently, saliva has been Methods: We performed a two-centre, in-clinic suggested as an alternative sample for DPD evaluation, (confinement), forced-switching, randomised potentially presenting higher stability. Objective: To controlled clinical study in Fukuoka-Japan evaluate two DPD phenotyping strategies, using (UMIN000024988, ISRCTN14301360). Baseline endogenous UH2/U ratios in both plasma and saliva, levels of selected BoE to cigarette smoke toxicants in together with 5-fluorouracil area under the curve the exhaled breath and urine of 180 smokers were (AUC) as predictive tests for screening for severe compared to those seen when the smokers either chemotherapy related toxicity. Materials and Methods: remained smoking combustible cigarettes or switched Paired plasma and saliva samples were obtained from to using a mentholated or non-mentholated THP for 5 13 patients scheduled for 5-FU infusion treatment for days. The study was approved by a local IRB and was gastrointestinal cancer, before starting of treatment. U run in accordance with ICH-GCP. Subjects provided and UH2 concentrations were measured in both written informed consent prior to study participation matrices by LC-MS/MS and 5-FU by HPLC-DAD. and were deemed healthy following medical The 5-FU AUC was calculated by multiplying the examination and clinical laboratory screening. plasma concentration with the infusion duration. Smoking status was verified by exhaled CO and Adverse effects after the first chemotherapy cycle were urinary cotinine measurements. Results: 180 subjects classified according to NCI-CTCAE Results: The completed the study. Subjects who switched from [UH2]/[U] metabolic ratios were highly different smoking to exclusive use of the glo™ device (n=30) among patients, from 0.62 to 9.19 in saliva and 0.82 to for 5 days showed reductions in levels of exhaled 34.4 in plasma. 5-FU AUC values ranged from 11.8 to carbon monoxide and a range of urinary BoEs, 37.3 mg.h/L and were under the therapeutic range of compared to their levels at baseline. In contrast, the 20-25 mg.h/L in 84% of the patients. The saliva continued smoking group (n=30) showed little change demonstrated higher correlation to 5-FU AUC in in BoE levels throughout the study. Results & comparison to plasma (rs=-0.827, P<0.05 vs rs=-0.198 Conclusion: In this human study, we observed P<0.05). Only one patient developed severe toxicity reductions in BoE in subjects who switched to using (grade 3 neutropenia), this patient had UH2/U ratio in the gloTM THP. These findings are in accordance with saliva bellow the cut-off for prediction of toxicity the lower levels of toxicants measured in the machine previously proposed (1.16). Conclusions: Most of the emissions from the gloTM THP. Future studies will patients had 5-FU systemic exposure bellow the examine whether these reductions in exposure are suggested therapeutic range, leading to low incidence preserved for prolonged, ambulatory periods and also of severe toxicity in this small group of patients. The whether these lead to any changes in health effects evaluation of the metabolic ratio [UH2]/[U] is a indicate. promising functional test of DPD, presenting a high correlation to 5-FU AUC. Financial support / Acknowledgments: FAPERGS CL 19 - PRELIMINARY STUDY ON THE EVALUATION OF ENDOGENOUS PLASMA AND SALIVARY URACIL TO CL 20 - EVALUATION OF THE IN VITRO DIHYDROURACIL METABOLIC RATIOS AS EFFICIENCY CAPTURE OF LIPOSSOMES PREDICTIVE TESTS TO 5-FLUOROURACIL WITH TRANSMEMBRANE PH GRADIENTES AUC AND THE OCCURRENCE OF SEVERE IN THE TREATMENT OF COCAINE CHEMOTHERAPY RELATED TOXICITY OVERDOSE

NETO, O. J. V. 1; RAYMUNDO, S.1;TEGNER, M. 1; FERNANDES, S. R 1; CINTRA, E. R 1; MARTINS, FRANZOI, M. A. 2; SCHWARTSMANN, G. 2; E. L. 1. LINDEN, R. 1; ANTUNES, M. V. 1. 1. Universidade Federal de Goiás, Goiânia – GO. 1 Universidade FEEVALE, Novo Hamburgo – RS, Introduction: According to the World Health Brasil ; 2 Hospital de Clínicas de Porto Alegre, Porto Organization the use of illicit drug is prevalent Alegre – RS, Brasil. worldwide and it is one of the 20 largest risk factors. Cocaine (benzoylmethylecgonine) is an alkaloid Introduction: Fluoropyrimidine toxicity has been extracted from coca leaves (Erythroxylon coca). Even related to a deficiency on its metabolic clearance known the physiological effects of cocaine, there is no mediated by dihydropyrimidine dehydrogenase (DPD) effective pharmacotherapy to treat addiction or ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 56 overdose. Looking for new treatments, the liposomes Introduction: Brazil leads the world ranking as the with transmembrane pH gradients known as "Drugs country that imports most pesticides in the world. scavengers" can be a potential strategy. It behaves Acute intoxications are the most visible face of the sequestering the drug into the bloodstream, making it impact of these products on health. Hence, the need for less available to peripheral tissues, effectively tools and instruments that can corroborate for a global reversing the effects of overdose. Aim: to develop, view of the impact of pesticides from the public health characterize and evaluate the in vitro performance of point of view. There are models in a health information liposomes with a transmembrane pH gradient for the system such as the Notification of Injury Information treatment of cocaine overdose cases. Material and System - SINAN and the National Information System methods: liposomes composed of different lipids (egg on Sanitary Surveillance - SINAVISA. Objective: To phosphatidylcholine, hydrogenated soy carry out a comparative study between the information phosphatidylcholine and DSPC) were prepared by the systems for notification of aggravated intoxication by lipid film hydration / extrusion method, such hydration agricultural pesticide between SINAN and SINAVISA being carried out with citrate buffer and sodium sulfate in the period from 2014 to 2016. Material and Method: (250 mM; pH 3.0). The pH gradient was obtained A documentary, descriptive and retrospective study replacing the external buffer for isotonic buffered was carried out. Data were collected from reports of saline solution (HBS) at pH 7.4 using dialysis the Toxicological Information Center of Goiás and the technique. Liposomes were characterized according to Reference Center on Workers' Health. Median and the size and zeta potential. In vitro tests to evaluate interquartile range or absolute and relative frequencies liposomal capture capacity (CC) were performed using were used. A chi-square test was performed with Yates lidocaine as proof of concept. The liposomes were correction for comparison of proportions and the Mann incubated at 37°C under horizontal shaking in HBS Whitney test for mediansa comparison, both with solution for 1 hour in the presence of lidocaine (1 mM) significance level of 5%. The STATA® software with a drug / lipid molar ratio of 0.3. At the end of the version 12.0 was used in this analysis. Results: In the test the free drug was separated from the liposomes by years 2014-2016, 826 notifications were made by ultracentrifugation and quantified by HPLC. Results: SINAVISA and 709 by SINAN. There were reports of Liposomes were obtained with 120 to 180 nm and zeta 407 tokens. The total number of cases in the evaluated potential of -3.0 mV. The CC was shown to be years should be 1128 cases. There was no statistical dependent on the lipid and the hydration medium used. difference between SINAN and SINAVISA (p> 0.05) The formulation composed by DSPC and sodium in relation to the year of notification, age and gender. citrate buffer presented better results for CC about 0.18 However, significant differences (p <0.05) were ± 0.01 (64.6%) after 1 hour of incubation. Conclusion: evidenced for most of the analyzed variables, as type initial results demonstrated that liposomes with a of herbicide pesticide. Conclusion: In the present transmembrane pH gradient showed good in vitro CC study, the most effective system was SINAVISA, due and are considered to be promising agents in the to the greater registration of notifications, more data treatment of drug overdose. quantities more specific from the toxicological point of Financial support / Acknowledgments: FAPEG; view. However, none were considered gold standard. CNPQ. Therefore, we consider the need for a more adequate tool, in addition to greater cooperation on the part of the professionals responsible for the notifications, in CL 21 - A COMPARATIVE STUDY BETWEEN the public scenario of health surveillance. THE NOTIFICATION OF INJURY Financial support / Acknowledgments: Secretaria INFORMATION SYSTEM - SINAN AND Estadual de Saúde de Goiás. NATIONAL SANITARY SURVEILLANCE SYSTEM - SINAVISA AT THE HEALTH SURVEILLANCE SUPERINTENDENCE OF THE STATE OF GOIÁS - SUVISA. RETROSPECTIVE STUDY FOR THE PERIOD FROM 2014 TO 2016

NAKANO, S.M. S.1; DINIZ, D.D.1; CARDOSO, H.A.1; LIRA, A.D.1; ROCHA, H.C.1; FREITAS, J.V.1; BRASIL, M.F.S.1; ARAÚJO, A.G.1; MORAES, M.V.1, DAMANDO, A.G1; SILVA, F.C.M.O.1

1. Secretaria Estadual de Saúde de Goiás

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CL 22 - AN EPIDEMIOLOGICAL STUDY OF Financial support / Acknowledgments: PESTICIDE POISONING NOTIFIED BY Superintendência de Vigilância em Saúde /Secretaria CENTER OF INFORMATION Estadual de Saúde de Goiás. TOXICOLOGICAL OF GOIAS IN THE PERIOD 2013- 2016 CL 23 - FIRST DESCRIPTION AND NAKANO, S.M.S.1; DINIZ, D.D.1; BRASIL, EVALUATION OF SNPS IN THE ADH AND M.F.S.1; FREITAS, J.V.1; ARAÚJO,A.G.1; ALDH GENES IN A POPULATION SAMPLE OF MORAES, M.V.1, DAMANDO, A.G.1; CARDOSO, ALCOHOLICS IN THE CENTRAL-WEST H.A.1; LIRA, A.D.1; ROCHA, H.C.1; SILVA, REGION OF BRAZIL F.C.M.O.1 TEIXEIRA, T. M.; GOVEIA R M¹; DELLEON H S¹; 1. Secretaria Estadual de Saúde de Goiás LIMA A P¹; FARIA R S¹; SILVEIRA-LACERDA EP¹; RIBOLLA P E M²; ALONSO D P²; SILVA Introduction: Brazil is the largest consumer of MELO D³; ALVES A A³. pesticides in the world. Acute intoxication are the most visible face of the impact of these products on health. 1- Universidade Federal de Goiás – GO, Brasil; 2 – A lot of farmers are estimated to be intoxicated Universidade Federal de Goiás, GO – Brasil; 3 – annually worldwide, mostly in developing countries, Universidade Estadual Paulista de Botucatu, SP, where the highest number of deaths from human Brasil. exposure to these agents is observed. Objective: The aim of this study were to carry out an epidemiological Alcoholism is a disease defined as a physical and / or survey of pesticide poisoning registered in the psychic state where the individual's need to ingest the Toxicological Information of Goias, Brazil (CIT/GO), drug constantly is characterized. Consequently, with clinical and epidemiological evaluation from alcoholism appears to be a serious public health 2014-2016. Material and Method: A cross-sectional problem. Some individuals metabolize alcohol better descriptive study using human poising by pesticides than others. The elimination of ethanol occurs by data provided by CIT/GO. There was analyzed oxidation of the same in acetaldehyde and thereafter in descriptive of the data in 826 files of the acetic acid and water. The reactions are catalyzed by notification/investigation and it used median and enzymes Alcohol dehydrogenase (ADH) and aldehyde interquartil interval or absolutes and relatives dehydrogenase (ALDH). The objective of this study is frequencies. The software STATA® version 12.0 was to characterize polymorphisms in alcohol users in the used in this study. Results: The most of pesticide’s region of Goias to establish a correlation with notifications poisoning for the study was by CIT/GO in alcoholism. The genotyping of the student population the 2014 year was 343 notifications. Most of them of the Psychosocial and Drug Attention System (n=78) were adults (83.41%), in men, who live in rural areas and Alcohol Recovery Center (n=21) was carried out and employees registered or not (94.42%). The most by the TaqMAn® SNP Genotyping Assays system and frequently reported conditions were occupational a statistical analysis performed through the GENEPOP attempt (48.45%) and suicides (31.96%). The main software. The results of the analyzes show that the chemical group (83.82%), were herbicide (48.36%) e ADH4 * 2 polymorphic variant is associated with the insecticide (36.6%). There was clinical manifestation development of alcoholism in the patients, in contrast, in 95.16%, and the most frequently was: vomit as polymorphic variants ADH1B * 2, ADH1C * 2 and (39.59%), headache (36.36%), nausea (35.23%) e ALDH2 * 2 are associated with the protection of the unwell (27.27%). In the most of the cases it was individual against alcoholism. In addition, linkage considered moderate intensity (19.01%) e severe disequilibrium analysis between ADH1B * 2 and (5.93%), and in the most of them occurred clinical ADH1C * 2 was performed, in which four experienced diagnostic (98.55%). In the follow-up, evolution more haplotypes (CA, CC, TC and TA) were performed. In common was the cure without sequel (94.19%), with the Odds Ratio analysis, the most frequent genotypes mortality in 2.66% of the cases. Conclusion: The most (found in the control population) were CT / TT, so that notifications of pesticide poisoning for the study was patients with any of the six haplotypes presented a occupational and with low intensity in acute lower susceptibility to the development of alcohol intoxication. However, these results indicate that the dependence. population in question presents vulnerabilities. We Financial support / Acknowledgments: FAPEG consider the need the better control the use of the pesticides and diagnosis of poisoning by pesticides, with aim the degree the risk of this great grievance the relevance in the public health.

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CL 24 - CONTROL OF WATER QUALITY: DILIGENCE IN THE HEALTH OF THE DIALYTIC PATIENT

BORGES, L. A. G. 1; CURI, M. 1; COUTO, M. B. 1; NOVAES, V. R. 1; SOUZA FILHO, F. R. 1.

1. Universidade de Rio Verde, Rio Verde - GO, Brasil. Introduction: The most used treatment of Chronic kidney disease is hemodialysis, that consists in an extracorporeal circulation of blood into tubes, where the residues and the excess of liquids are filtered, and it’s constantly bathed by an appropriate electrolyte solution. Even with the increasing sophistication of this procedure, pyrogenic reactions still occur due to contamination of the water used. Objectives: Analyze the scientific production about the quality of water used to treat patients who undergo hemodialysis. Materials and methods: Twenty articles selected from the Virtual Health Library. The selection criteria: to be easily accessible, to be published until March 2014, in Portuguese, English and Spanish. Results: Pyrogenic reactions during dialysis are less frequent with modern water treatment systems. When they occur, contamination of the water is suggested, which is higher in the high flow dialyses, in which the retrograde filtration of the dialysate takes place. The recommendations about water for dialysate or for reuse are: a monthly bacterial count of less than 200 colony forming units (CFU)/ml and the level of action Is 50 CFU/ml. The endotoxins are restricted to 0.25 endotoxin units/ml. When a patient presents pyrogenic reactions during hemodialysis, samples should be immediately collected and sent for analysis. In 2011, a study in Brazil revealed that the most common bacteria in dialysis water are Pseudomonas aeruginosa, Klebsiella spp., Staphylococcus aureus. And the most prevalent fungi are: Penicillium spp., Aspergillus spp., Trichoderma spp.. Bacterial infections are the most common and fungi are relatively rare, but both are related to high morbidity and mortality. The gold disinfection standard is hot water, exceeding 80 degrees Celsius. It is an excellent procedure for eliminating bacterial contamination without the side effects of chemical germicides. Conclusion: Because of the contamination of the water used in hemodialysis and because these pacients are more susceptible to infections, we reinforce the importance of rigor about the use of protocols in the analysis of the water and the processes of desinfection and care general. Financial support / Acknowledgments: Liga Acadêmica de Nefrologia da Faculdade de Medicina de Rio Verde.

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EN 01 - EFFECTS OF PICOXYSTROBIN AND biomass and genotoxicity by means of micronuclei and CIPROCONAZOLE ON BULLFROG nuclear abnormalities. TADPOLES (LITHOBATES CATESBEIANUS) Financial support / Acknowledgments: FAPESP BY MELANOMAGNETIC ANALYSIS Processo 2016/20575-6

ADRIANA SACIOTO MARCANTONIO; CLÁUDIA MARIS FERREIRA; SÉRGIO HENRIQUE EN 02 - RACTOPAMINE EXPOSURE CAUSES CANELLO SCHALCH; FERNANDA MENEZES OXIDATIVE STRESS IN ZEBRAFISH (DANIO FRANÇA RERIO)

1. APTA - Agência Paulista de Tecnologia dos SACHETT, A. 1; BEVILAQUA, F. 1; Agronegócios - Polo Regional do Vale do Paraíba, CHITOLINA,R. 1; GARBINATO, C.L. 1; Pindamonhangaba - SP; 2. - Instituto de Pesca, São GASPARETTO, H.M. 1; CONTERATO, G.M.M 2; Paulo - SP; 3. - APTA - Agência Paulista de SIEBEL, A.M. 1. Tecnologia dos Agronegócios - Polo Regional do Vale do Paraíba, Pindamonhangaba - SP; 4. APTA - 1.Universidade Comunitária da Região de Chapecó, Agência Paulista de Tecnologia dos Agronegócios - Chapecó – SC, Brasil; 2. Universidade Federal de Polo Regional do Vale do Paraíba, Pindamonhangaba - Santa Catarina, Campos Novos - SC, Brasil. SP. Introduction: The presence of veterinary drugs in the Introduction The use of biological indicators has environment, especially in water bodies is a current increased in recent years in order to investigate reality. Consequently, compounds that produce environmental pollution, and the aquatic environment adverse effects may reach nontarget organisms. is highly vulnerable, with agriculture being a major Ractopamine is a β2-agonists used as feed additive to human activity contributing to increased pollution, promote growth rates, increase muscle mass, and Due to the use of pesticides. These substances, among reduce fat deposition and feed consumption. This environmental pollutants, have received recent veterinary drug has been detected in wastewater, attention for their potential in changing populations groundwater agricultural and in effluent-dominated and the dynamism between communities. Biomarkers streams, but the effect of this on exposed organisms is can help assess the health status of amphibian unknown. Objective: We evaluated the effects of populations, acting as final and sublethal parameters of environmentally relevant concentrations of intoxication. Due to the need to establish biomarkers ractopamine on oxidative stress parameters in with greater sensitivity and precision the analysis of zebrafish (Danio rerio) brain. Materials and Methods: the extracutaneous pigmentary system and specifically Adult wild-type zebrafish at both sex were exposed for in hematopoietic organs such as spleen and liver, a period of 7 days to ractopamine hydrochloride in where the melanomacrophages (MMCs) will reside, concentrations at 0.85, 8.5 and 85 µg/L. The control will be tested. Goal To evaluate the effect of group was maintained under the same conditions of the APROACH ® PRIMA fungicide on cutaneous and experimental group. Subsequently, animals were hepatic pigmentation, as well as classical sublethal and euthanized and brains were collected to assess the lethal effects on tadpoles of L. catesbeianus. Material oxidative stress parameters. The data were analyzed and methods The commercial product Aproach® using one-way ANOVA followed by Tukey's post hoc Prima, which, in its toxicological classification, test. Results: Fish exposed to ractopamine at 0.85, 8.5 belongs to class III, being moderately toxic and in and 85 µg/L showed a significant decrease in non- relation to the classification of the environmental protein reduced sulfhydryl (NPSH) levels (p<0.05) hazard potential, belongs to class II, which is very when compared to the control group. Ractopamine also dangerous to the environment, according to the caused a significant decrease in superoxide dismutase product. This product was chosen because of its use in activity (p<0.05) at the highest concentration (85 µg/L) the culture of flooded rice, which is the main crop of and an increase in catalase activity (p<0.05) at the Paraíba Valley region and has a great interface concentration of 0.85 µg/L in comparison with control with the aquatic environment. The test organisms will group. The NPSH reduction and alterations in the be bullfrog tadpoles, divided into 6 treatments and 3 activity of antioxidant enzymes suggest that replicates, for initial determination of LC50-96h and ractopamine may alter the antioxidant status of later micronucleus test and extracutaneous zebrafish brain, even though there was no alteration in pigmentation. Results To relate the possible effects on thiobarbituric acid reactive substances (TBARS) melanomacro- gages with the classical toxicological levels. Conclusions: In summary, our results suggest and lethal toxic effects of APROACH® PRIMA that ractopamine at concentrations already found in exposure, such as mortality, behavior, growth, surface water may change the antioxidant status of development and presence of abnormalities, tadpoles zebrafish brain. In addition, NPSH reduction is likely ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 61 to be an early event of environmental exposure to subgroup were rs1800668 CC and rs1050450 CC ractopamine in zebrafish. Further studies are needed to (45.3% and 54.6%, respectively). A statistically elucidate the biological meaning of these effects as significant association was found between well as the underlying mechanisms. polymorphism rs1800668 and an increase in Hg levels Financial support / Acknowledgments: CAPES - ( 2 = 4.11; p = 0.043). Conclusion: The results FAPESC / UNOCHAPECO showed that polymorphism rs1800668 is associated with an increase in Hg levels, but more studies are needed to explain the association found in this EN 03 - EFFECT OF GPX1 GENE ON population environmentally exposed to Hg. MERCURY LEVELS IN A LOW AMAZON Financial support / Acknowledgments: Fapespa, POPULATION Capes, Cnpq

NEVES A.C 2,8; MENESES H.N.M.1,2; MENESES A.A.M 3,4,5; SAMPAIO DA SILVA D.6,7; EN 04 - STANDARTIZATION OF IN VIVO AND RODRIGUES L.R.R 2,3,4. IN VITRO ASSAYS FOR AROMATASE INHIBITORS – PRELIMINARY RESULTS 1 Instituto de Saúde Coletiva, Universidade Federal do Oeste do Pará, Santarém –PA Brasil; 2 Laboratório de BOARETO, A. C. 1; MULLER, J. C. 2; GOMES, C. Genética e Biodiversidade, Universidade Federal do 3; SILVA, J. G. 4; GRECHI, N. 1; ANDREOTTI, B. Oeste do Pará, Santarém - PA Brasil; 3 Programa de 1; DALSENTER, P. D. 1. Pós Graduação em Recursos Naturais da Amazônia, Universidade Federal do Oeste do Pará, Santarém –PA 1. Universidade Federal do Paraná, Curitiba - PR, Brasil ; 4 Programa de Pós Graduação Sociedade Brasil; 2. Faculdades Pequeno Príncipe, Curitiba - PR, Natureza e Desenvolvimento, Universidade Federal do Brasil; 3. BASF, Ludwigshafen - Alemanha; 4. Oeste do Pará, Santarém –PA Brasil ; 5 Laboratório de Universidade Tecnológica Federal do Paraná, Curitiba Inteligência Computacional, Universidade Federal do - PR, Brasil. Oeste do Pará, Santarém –PA Brasil ; 6 Centro de Formação Interdisciplinar, Universidade Federal do Introduction: The aromatase enzyme has an important Oeste do Pará, Santarém –PA Brasil ; 7 Programa de role in reproductive processes and it is involved in the Pós Graduação Sociedade, Ambiente e Qualidade de development of diseases with high levels of incidence vida, Universidade Federal do Oeste do Pará, Santarém and metastatic potential. EPA recommend an in vitro –PA Brasil ; 8 Instituto de Biodiversidade e Florestas, test for screening of aromatase inhibitors however, Universidade Federal do Oeste do Pará, Santarém- PA there is no specific in vivo test to evaluate changes in Brasil. the aromatase. Objective: This project aims to standardize an in vivo screening test capable of Introduction: In the Amazon region, mercury (Hg) is identifying substances that may inhibit the aromatase abundant and can be considered a xenobiotic in the enzyme. Materials and Methods: Immature female human body. Its methylated form (MeHg) is Wistar rats (21 days old) were treated daily, during bioaccumulative, causing a decrease in the activity of five days. Nine experimental groups, with 12 animals the enzymes of the antioxidant system. The enzyme per group, were used: vehicle control (canola oil); Glutathione Peroxidase (GPx1) protects cells from estradiol 10 µg/mL (positive control for estrogenicity); oxidative damage by neutralizing free radicals. testosterone 3.5 mg/kg + flutamide 5 mg/kg (aromatase Objective: Verify the frequency of polymorphisms in activity control in doses with uterotrophic effect by the the GPx1 gene and its relation with Hg levels in a aromatase conversion of testosterone into estrogen); Lower Amazon population. Metodology: The Hg three different doses of exemestane 0.36/3,6/36 mg/kg levels of 144 individuals were quantified. Genotyping and anastrozol 0.015/0.15/1.5 mg/kg (known of the GPx1 gene rs1800668 C> T and rs1050450 C> aromatase inhibitors) associated with testosterone + T polymorphisms was performed by the High flutamide. On the last day of treatment, the animals Resolution Melting (HRM) technique. The Kruskal- were weighed and euthanized. Uteri were removed and Wallis test was applied to verify the effect of GPX1 full and empty uterine weights were measured. All gene polymorphisms on Hg levels. Results Of the 144 experiments were approved by the Ethics Committee individuals analyzed, genotype rs1800668 CT and on Animal Experimentation of Federal University of rs1050450 CC were the most frequent (38.6% and Parana (no 798). Results: The uteri weight from the 51.1%, respectively). Regarding the level of Hg, the rats treated with anastrozol+testosterone+flutamide highest levels were found among individuals with and with exemestane 36 genotypes rs1800668 CT and rs1050450 CC (44.3% mg/kg+testosterone+flutamide were not different from and 51.1%, respectively). Regarding fish consumption, the vehicle control group, and were different from the the most frequent genotypes in the high frequency uteri weight of the testosterone+flutamide group. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 62

These results show that the inhibition of the aromatase <5 µg/dL, CDC´s attention level. Children’s IQ mean was enough to prevent the conversion of testosterone was 76 (43–116), with 58% below 80 points, clinical into estradiol. Conclusion: This study showed that is cut-off for challenged children. Similarly, the maternal possible the development of an in vivo screening test mean IQ was 68 (43– 108), with 77% below 80. to evaluate aromatase inhibitors using immatures Despite this low exposure level, a very significant female rats. Further studies, as the determination of the negative association was observed between LogPbB aromatase activity and expression, and the levels of and children´s IQ (β-coefficient -8.58; 95%CI -14.2 to estrogen and testosterone, are necessary to validate the -2.9; p=0.003) independent of age and maternal IQ. No test. sex difference was observed. Conclusions: Our data Financial support / Acknowledgments: Fundação shows that very low Pb exposure level may induce Araucária. CAPES. negative effects on children’s intellectual performance. Financial support / Acknowledgments: Conselho Nacional de Desenvolvimento Científico e EN 05 - SCHOOL-AGED CHILDREN Tecnológico (CNPq); Fundação de Apoio a Pesquisa EXPOSURE TO LOW LEAD LEVELS AND THE do Estado da Bahia(FAPESB). ASSOCIATION WITH INTELLECTUAL FUNCTION EN 06 - TOXICOLOGICAL AND QUALITATIVE ANJOS, A. L. S. 1; RODRIGUES, J. L. G. 1; ANALYSIS OF THE PRESENCE OF HEAVY BANDEIRA, M. J. 1; SANTOS, N. R. 1; SANTOS, S. METALS IN THE ATMOSPHERIC C. 1; CARVALHO, C.F. 2; LIMA, C.S. 2; BONFIM, PARTICULAR MATERIAL (MP10) OF THE R.S. 2; ARAÚJO, C.F.S. 3. MENEZES-FILHO, J.A. MUNICIPALITY OF JI-PARANÁ (RO) 2. OLIVEIRA, C. E. F. 1; SANTOS, C. M. R. 1; 1. Laboratório de Toxicologia, Faculdade de farmácia, AZEVEDO, M. S. 2; SOUTO, L. F.. L. 1; VALE. L. Universidade Federal da Bahia, Salvador - BA, Brasil; A. S. 1; SOUZA, A. O. 1. 2. Instituto de Psicologia, Universidade Federal da Bahia, Salvador - BA, Brasil; 3. Escola Nacional de 1 Instituto Federal de Rondônia, Ji-Paraná - Rondônia; Saúde Pública, Fundação Osvaldo Cruz, Rio de Universidade Federal de Rondônia, Porto Velho - Janeiro-RJ, Brasil. Rondônia.

Introduction: Lead (Pb) is a toxic heavy metal Introduction - Air pollution is an environment problem ubiquitous in the environment. It has several effects on in different countries around the world including the human organism, especially to the nervous system Amazon region due increases of anthropogenic (NS) of developing children. Studies have been shown activities, outbreaks of fires and the local vehicular that Pb can affect children’s NS, causing intellectual fleet however, studies related to the atmospheric deficit, hyperactivity and attention problems. Several quality of the northern Brazilian region are still not studies have demonstrated that for urban children, dust widespread. One of pollutants of air is the particulate may represent the principal exposure source, especially matter which can be composed by dust, organic for young toddlers due to the mouthing habit. compounds and heavy metals which shows high levels Objective: To evaluate de association between blood of toxicity to human health, already being related to lead levels (PbB) and intellectual deficit in school- the development of cardiovascular diseases and cancer. aged children from Simões Filho, Bahia. Materials and Objective - Therefore, considering the high levels of Methods: School-aged children (7-12 years), 110 boys particulate matter in the northern region, the natural and 109 girls, studying in four different schools were presence of different toxic pollutants to health and the evaluated. Pb exposure was measured in school settled absence of studies on the quality of the atmospheric dust and in children´s blood by graphite furnace atomic environment of several cities of north region such as absorption spectrometry. Children´s IQ were estimated Ji-Paraná-RO, this study aimed to investigated the by Wechsler Abbreviated Scale for Intelligence mutagenic potential and the presence of heavy metals (WASI) using the subtests of vocabulary and reasoning in the particulate matter in Ji-Paraná-RO by analyzes matrix. Maternal or main caregiver’s IQ was also of the increase in the incidence of micronucleus of evaluated using the adult version of WASI. Data were Allium cepa after exposure to particulate matter and analyzed using SPSS version 22, with significance heavy metal qualitative analysis by X-Ray level of p<0.05. Results: Pd in settled dust in interior Fluorescence (FRX). Material and Methods - The environments varied 44.5 to 73.4 µg/m2/30 days lower samples are collected on polycarbonate membrane by than the USEPA’s floor dust standard (131.2 µg/m2). for seven days in three main avenues of the cities by Children´s PbB median (P25-P75) was 1.2 (0.2 – 15.6) natural depositing through the air current. To µg/dL. From the 219 children, 98.2% had PbB levels mutagenic analyses, samples were extracted in water ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 63 using ultrasound for 15 minutes and submitted to presence of different metals in the sample may have Allium cepa methodology and to heavy metal contributed to the results obtained. The vinasse–soil qualitative analyses were development FRX Analysis. interaction is also a factor that influences toxicity. Results - Our results showing increases of mutagenic According to the results, the combined analysis of evidences (increases of micronucleus) at the main different parameters of the A. cepa test system showed intersections of the avenues and it was also noted that high dilutions of orange vinasse exhibited several metals (Cu, Fe, Co and Zn) of toxicological phytotoxicity and genotoxicity for this organism when profile already reported in the literature and they are applied in soil., so caution is suggested in the disposal similar to other in urban centers. Conclusion - These of this effluent into the environment. Key words: citrus air pollution evidences may be related to rates of stillage, fertirrigation, phytotoxicity, chromosomal respiratory diseases in the Rondonia-Brazil in the last aberration, micronucleus, mutagenic potential. years and shows the current dangers of atmospheric Financial support / Acknowledgments: pollution that already affect the Brazilian Amazon. FAPESP, process no. 2014/17998-7 and 2012/50197-2 Financial support / Acknowledgments: CNPq

EN 08 - EVALUATION OF THE TOXIC EN 07 - GENOTOXIC AND PHYTOTOXIC POTENTIAL OF PHYTOREMEDIATED EVALUATION OF ORANGE VINASSE VINASSE USING THE RHINOCRICUS APPLIED IN SOIL PADBERGI (DIPLOPODA) FAT BODY

GARCIA, C. F. H. 1; SOUZA, R. B. 1; DE SOUZA, GARCIA, C. F. H. 1; SOUZA, R. B. 1; DE SOUZA, C. P. 1; ANGELIS, D. F. 1; LEITE, D. A. N. O. 1; C. P. 1; MARCATO, A. C. C. 1; FONTANETTI, C. S. FONTANETTI, C. S. 1. 1.

1. UNESP – São Paulo State University, Biology 1. UNESP – São Paulo State University, Biology Department, Biosciences Institute, Rio Claro, SP, Department, Biosciences Institute, Rio Claro, SP, Brazil Brazil.

Constant and inappropriate use of pesticides, Along with the high production of ethanol, the residues agricultural fertilizers and low-quality irrigation water, generated by the agro-ethanol industry, such as as well as the indiscriminate disposal of industrial or vinasse, also increased. As an alternative to its domestic waste can cause the accumulation of harmful reutilization, the fertigation in sugar cane crops is the substances in soil. These substances can be toxic to most used. However, its use as fertilizer has generated plants and animals, including humans. Vinasse is one concern due to adverse effects in the environment. So, of several residues that have been widely released in many technologies, such as wetland systems, have soil and, for this reason, have gained attention from the emerged with the aim of treating vinasse and reducing scientific community. This effluent is a by-product of its toxic potential. This study aimed to evaluate the alcohol production obtained from several raw vinasse toxicity after phytoremediation by water materials, such as orange, corn, sugar cane and beet, hyacinth in a wetland system. Histological tests were among others. Sugar cane vinasse has been widely performed in order to analyze semi-quantitatively used in fertirrigation and several studies demonstrated alterations in the perivisceral fat body, organ with its toxicity to soil and water organisms. Orange vinasse important metabolic function, of diplopods presents similar characteristics to sugar cane vinasse, (Rhinocricus padbergi) exposed to soil with the so it may also be used as a potential fertilizer in the phytoremediated vinasse. For the preparation of the agriculture activity. The objective of this study was to bioassays, two treatments were set up: one control and evaluate the toxicity of orange vinasse, when applied one with soil containing 100% of the phytoremediated to soil using the Allium cepa test system. The vinasse dosage according to CETESB standards. evaluation was performed by means of germination, Fifteen animals were randomly assigned to each root growth, genotoxic and mutagenic parameters. The terrarium, where they were exposed for periods of 21 LD50 and ½ LD50, defined in the germination test, and 42 days. The experiment was carried out in were used for genotoxicity tests. According to the triplicate. After each exposure period, three individuals regression curve based on the mean values for root from each terrarium were anesthetized and dissected growth, the estimated LD50 was 47.43%. for the removal of the perivisceral fat body. Slides Phytotoxicity was observed in dilutions above 40%, by were prepared and stained with hematoxylin-eosin. means of germination and root growth tests. The control in both periods of exposure showed the Significant chromosomal alterations were observed at histological pattern described for the species. After 21 47.43% (LD50) dilution and mutagenicity was not days of exposure to phytoremediate vinasse, a small observed. Low pH, high BOD, high COD and the increase in the quantity of spherocrystal, trophocytes ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 64 and oenocytes was observed, but without alteration in Financial support / Acknowledgments: CNPq - their morphology. The animals exposed to 42 days also Conselho Nacional de Desenvolvimento Científico e presented an increase in quantity of spherocrystals and Tecnológico. oenocytes, however frequent regions with cytoplasmic disorganization and loss of cellular limit were observed. According to the results, although the EN 10 - RISK ASSESSMENT OF EMERGING vinasse treatment reduced some of the physico- CONTAMINANTS IN MAIN WATER INTAKES chemical parameters, a chronic exposure can alter USED IN FEDERAL DISTRICT, BRAZIL some characteristics related to the toxicity in the diplopods fat body. Thus further studies are needed to CAVALCANTI, C. M. P. 1; SODRÉ, F. F. 2 better understand the effect of the phytoremediated vinasse on organisms. 1. Universidade de Brasília, Brasília - DF, Brasil, Financial support / Acknowledgments: FAPESP, CAESB - Companhia de Saneamento Ambiental do process nº. 2016/19817-5 Distrito Federal, Brasília - DF, Brasil;2. Universidade de Brasília, Brasília - DF, Brasil.

EN 09 - PRIORITIZATION OF CONTAMINANTS IN DRINKING WATER Introduction: Most of the environmental contaminants BASED ON TOXICITY AND ENVIRONMENTAL of emerging concern are not regulated, but need to be FATE ASSESSMENT BY IN SILICO MODELS: monitored to support safe use of water. This study AN INTEGRATED AND TRANSPARENT aimed to assess environmental and human health risks RANKING arising from the presence of selected emerging contaminants in the main sources of water from the SANTOS, C. E. M.1; NARDOCCI, A. C.1 Brazilian Federal District. Material and Methods: Measured environmental concentrations (MEC) of 1. Faculdade de Saúde Pública, Universidade de São atrazine (ATZ), caffeine (CAF), carbamazepine (CBZ) Paulo, São Paulo-SP, Brasil. and atenolol (ATN) were obtained by solid phase extraction followed by liquid chromatography-mass Introduction: Occurrence of micropollutants in water spectrometry. Environmental risks were estimated worries the scientific community, regulators and through predicted no-effect concentrations (PNEC). population. Many recent studies have suggested Water quality criteria (WQC) for human consumption different criteria for the definition of priority were estimated according WHO guidelines. Both molecules, . Methods: In this work an assessment MEC/PNEC and MEC/WQC (considering no removal environmental fate and the potential of toxicity of in water treatment) rations were compared. Values active pharmaceutical ingredients (APIs) was provided above 1.0 indicate potential risk, whereas values below by in silico models and data review. A visual and 0.1 indicate no risk. Intermediate values indicate transparent Toxicological Prioritization Index (ToxPi) possible risks as well as the need for further is proposed based on in silico, in chemico, in vitro and ecotoxicological data. Results: No risks were predicted in vivo data. APIs considered relevant were selected for Santa Maria and Descoberto lakes, where according to specific inclusion criteria. Predictions maximum MEC for ATZ - CAF were 3.4 - 10.4 ng/L about environmental fate and toxicity were made and 5.5 - 31.8 ng/, respectively. Corumbá Lake through in silico models of EPISuiteTM, PBT presented concentrations of 9.3, 149, and 8.5 ng/L for Profiler©, TEST© and QSAR Toolbox©. The ATZ, CAF and CBZ, respectively, as well as possible prioritization method of software ToxPi GUI was used environmental risk for carbamazepine. Maximum for a ranking based in environmental fate and human MEC in the water supplies at Paranoá Lake were 11.4 toxicity. The fate and hazard-based assessment ng/L (ATN), 10.3 ng/L (ATZ), 180 ng/L (CAF) and suggested higher concern APIs were metronidazole 20.8 ng/L (CBZ). In this lake, environmental risk was (ToxPi: 8.49), ethinylestradiol (ToxPi: 7.71) and evidenced for carbamazepine and possible risk for norethisterone enanthate (ToxPi: 6.28); and the lower caffeine. No human health risk was predicted in this concern were ibuprofen (ToxPi: 2.02) and work. Conclusion: Results show the need for further acetylsalicylic acid (ToxPi: 2.98). Different APIs may investigation regarding ecotoxicological effects for have high potential for environmental exposure and/or contaminants of emerging concern such as toxicity despite the lower volume of use or sale. pharmaceuticals, hormones and personal care products. Computational tools may be an important tool for Also, demonstrated the need for water quality environmental fate and toxicity assessments in order to standards to protect aquatic life. identify priority agents by transparent visual rankings Financial support / Acknowledgments: The authors and enable decision-making for these criteria. are grateful to CAESB for support and data provided and to FAPDF for financial support.

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EN 11 - ANEUGENICITY AND DNA EN 12 - STUDY OF THE VULNERABILITY OF FRAGMENTATION IN Allium cepa INDUCED RESIDENTS OF AN AREA UNDER INFLUENCE BY A COMMERCIAL FORMULATION OF 2,4-D OF THE INDUSTRIAL POLE OF CAMPOS HERBICIDE ELÍSEOS, BAIXADA FLUMINENSE, TO ENVIRONMENTAL CHEMICAL SOUZA, C. P. 1; SOUZA, R. B. 1; PALMIERI, M. J. CONTAMINANTS 2; ANDRADE-VIERA, L. F. 2; FONTANETTI, C. S. 1. BARATA-SILVA, C. 1; MOTA, C.L. 1; BRANCO, A.1; PAVESI, T. 1; MITRI, S. 1; MOREIRA, J.C. 1 1 Universidade Estadual Paulista (Unesp), Instituto de Biociências, Departamento de Biologia, Rio Claro, SP, 1.Fundação Oswaldo Cruz-RJ/RJ Brasil; 2 Universidade Federal de Lavras (UFLA), Departamento de Biologia, Lavras, MG, Brasil. INTRODUCTION: Campos Elíseos, a Duque de Caxias district in Rio de Janeiro, houses the largest The use of plant bioindicators to assess herbicide industrial complex in Latin America, including the genotoxicity is recommended in several studies. In this second largest petrochemical company in the country study the cytotoxicity and genotoxicity of a (REDUC. The main industrial segments in this region commercial formulation of the herbicide 2,4- are the chemical, petrochemical, metallurgical, gas and dichlorophenoxyacetic acid (2,4-D) in Allium cepa plastic sectors, indicating the great diversity of was evaluated through germination, root growth, products and inputs produced/consumed locally. chromosomal aberrations tests, TUNEL (TdT- Although Duque de Caxias had the 6th highest GDP in mediated deoxy-uracil nick end labeling) assay and the national ranking and the 2nd highest in the state, it DNA fragmentation by electrophoresis. Seeds of A. has an HDI of 0.753, the 52nd in the State and the 3rd cepa were germinated at the field concentration (0,548 worst among the neighbouring municipalities. μL /mL), half and a quarter of the herbicide field OBJECTIVE: The objective of the present study was concentration. The negative control (NC) used was to evaluate the vulnerability to chemical contaminant distilled water. The statistical analysis was performed environmental exposure of Campos Elíseos residents. by comparing the results of the treatments with the METHODOLOGY: The study was of the negative control; according to data distribution for observational descriptive exploratory type, carried out normality was performed the parametric test or the at CESTEH/FIOCRUZ. The study group consisted of nonparametric test. Most of the roots of all treatments adults over 18 years old, of both sexes and different presented a swelling just above the meristematic ages and races, who lived in Campos Elíseos for at region. According to the ANOVA/Tukey test (p≤0.05), least 3 months. The residents first answered a semi- all concentrations reduced germination and root structured questionnaire and a 4 mL aliquot of blood growth significantly when compared to NC. The was collected for hemogram. RESULTS: After an Kruskal Wallis/ Dunn test (p≤0.05) showed that explanation of the study, 190 residents volunteered to mitotic index also decreased significantly in the three participate, predominantly women (72%) with mean concentrations in relation to the NC; the two lowest age of 55 years old. All participants stated a lack of concentrations significantly increased morphological treated water and basic sanitation in the region. characteristics of the cell death process and the number Regarding pollution perception, about 98% of of cells with nuclear abnormalities, such as binucleated residents consider the region heavily polluted and 79% cells. The two lowest concentrations induced DNA reported some discomfort related to an unpleasant, gas- fragments around 300 base pairs. All concentrations like, odour. Regarding the blood analysis, about 33% showed TUNEL positive nuclei. Thus, the of the participants presented alterations characteristic concentrations tested of commercial 2,4-D herbicide of certain pathologies. The most frequent was anemia, were cytotoxic and aneugenic to the test organism followed by thrombocytopenia. The majority of used. hematological alterations were observed in participants Financial support / Acknowledgments: CAPES and residing in a region surrounded by industrial FAPESP proc. 2012/50197-2 complexes, indicating chemical contamination as a possible cause for these health outcomes. CONCLUSION: Although Duque de Caxias is a municipality with high tax revenues, basic investments in Campos Elíseos are not implemented, exposing the resident population to environmental contamination and resulting health problems. Financial support / Acknowledgments: FIOCRUZ/ CAPES

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EN 13 - APPLICABILITY OF MONOCYTE be used as an indicator of indoor air quality (IAQ) as a ACTIVATION TEST (MAT) AS AN complement to fungi count. TOXICOLOGICAL IN VITRO METHOD TO Financial support / Acknowledgments: ASSESS THE INDOOR AIR QUALITY (IAQ) IN CNPq/FIOCRUZ LABORATORY AREAS

SILVA, C. C.; CALDEIRA, C.1; VIEIRA, D.H. 1; EN 14 - USING QUANTITATIVE OLIVEIRA, C. B. N. 1, DELGADO, I. F. 3; WASTEWATER ANALYSIS IN A PRISION PRESGRAVE, O. A. F. 1; MORAES, A. M. L. 2. EFFLUENT TO MEASURE DAILY USAGE OF COCAINE 1. Department of Pharmacology and Toxicology, National Institute of Quality Control in Health PETEFFI, G. P. 1; BARBOSA, E. 1; HAHN, R. Z. 1; (INCQS), Oswaldo Cruz Foundation, Rio de Janeiro, LINDEN, R. 1 Brazil; 2. Laboratory of Taxonomy, Biochemistry and Bioprospection of Fungi, Oswaldo Cruz Institute 1 Laboratório de Análises Toxicológicas, Universidade (IOC) Oswaldo Cruz Foundation, Rio de Janeiro, Feevale, Novo Hamburgo, Rio Grande do Sul, Brasil. Brazil; 3. Vice Presidency of Education, Information and Communication Oswaldo Cruz Foundation, Rio de INTRODUCTION: Wastewater analysis is an Janeiro, Brazil. alternative method for monitoring population drug use. Additionally, ecotoxicological risks can be associated Introduction: The presence of bioaresols represent an to the presence of drugs and metabolites in the important risk factor to human health and contribute effluent, which is released in superficial waters. for airway diseases and health effects. The Act nº OBJECTIVE: To develop a HPLC-FL assay to 09/2003 drafted by National Health Surveillance measure cocaine (COC) and benzoylecgonine (BZE) Agency (Anvisa) which establishes technical levels in a local correctional facility effluent, guidelines for the "References standards of interior air estimating COC use and evaluating the associated quality in artificially conditioned environments for ecotoxicological risk. MATERIAL AND METHODS: public and collective use", recommend the fungi count Ecotoxicological risk was estimated as risk quotient as an indicator for microbiological contamination. (RQ) and classified as low (RQ< 0.1), medium However, this method only identifies live (0,1

OLIVEIRA-FILHO, E. C. 1,2; TALLARICO, L. F. 3,4 EN 15 - EFFECTS OF SILVER NANOPARTICLES ON THE AQUATIC SNAIL 1. Embrapa Cerrados, Planaltina - DF, Brasil; 2. BIOMPHALARIA GLABRATA UniCEUB, Brasília, DF, Brasil; 3. Instituto Butantan, São Paulo, SP, Brasil; 4. 4. Faculdade Nossa Cidade, OLIVEIRA-FILHO, E. C.1,2; CARVALHO, E. L.2 Estácio, Carapicuíba, SP, Brasil.

1. Embrapa Cerrados, Planaltina - DF, Brasil; 2 - Studies of the adverse effects of chemical compounds UniCEUB, Brasília, DF, Brasil. using gastropods have been fostered in several countries, mainly due to the importance of endemic The nanoscale of some materials, due to technological species with ecological significance, as these are development, has been applied in several areas of sensitive to a different range of pollutants, as well as science. Examples include silver nanoparticles the importance of establishing assays with organisms (AgNPs) that are part of textiles, materials to eliminate of benthic habit. The most common assays used in bacteria and smells from clothing, food packaging and worldwide ecotoxicological protocols are focused on other items where antimicrobial properties are acute and chronic toxicity, but the development of desirable. Due to their biocidal effect, it is necessary to techniques that assess the sublethal risks of pollutants evaluate the possible adverse effects of AgNPs in order in low concentrations, analysis of the effects on to know the limitations of the activities of these gametes, fertilization, reproduction and embryonic compounds, evaluating the hazards they may present to development are relevant and encouraged by human health and the environment. The fact that the environmental monitoring programs. In Brazil, besides aquatic snails are organisms that move on the substrate the more traditional test organisms in ecotoxicological makes them an interesting alternative to evaluate evaluations, such as microcrustaceans and fish, the particles of some materials in water, especially those most representative species of freshwater gastropods, that are poorly soluble and tend to precipitate in the with different approaches already established, are substrate, as well as several nanomaterials. Thus, this those from the Biomphalaria genus. This work work intends to investigate the effects of silver describes the main standardized tests and prospects for nanoparticles on the survival and reproduction of the regulatory processes with environmental agencies for benthic aquatic snail Biomphalaria glabrata. the evaluation of chemical substances and water Preliminary assays showed a 10-day LC50 of 18.57 samples using freshwater snails, which are important mg L-1. From this value, 40 adult individuals of B. tools to contribute to the conservation and monitoring glabrata with shell diameter between 12 and 15 mm of water pollution. were selected, and separated into 4 groups (10 Financial support / Acknowledgments: CNPq individuals per group), for individual exposure for 4 Processo: 478673-2012-8 weeks at dilutions of 1.0, 2.5 and 5 mg L-1AgNPs, in addition to the control group, without the presence of AgNPs. The results showed that there was significant inhibition in the number of eggs per individual in 2.5 mg L-1 in relation to the control. In the case of spawning per individual, there was significant inhibition at all concentrations. These findings show that silver nanoparticles present a NOEC < 1 mg L-1 for aquatic snails of B. glabrata species. Financial support / Acknowledgments: CNPq Processo: 552113/2011-5

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EN 17 - RUTHENIUM COMPLEXE (LGMC–69): EN 18 - EVALUATION OF TOXICITY AND A PROMISING LARVICIDAL AGENT AGAINST PHYSICOCHEMICAL PARAMETERS OF A AEDES AEGYPTI WATER BODY, PHARMACEUTICAL EFFLUENT RECEIVER NUNES, E. B.; ROMANO C.A.1; SILVA, I.G.1; SILVA, H.H.G.1; CARDOZO, S.P.1, BATISTA, A. CHEQUER, F.M.D. 1,2; SILVA, V.K.C.1; A. 2; SILVEIRA-LACERDA, E.P. 1. MOREIRA, T.F.M.1; ANASTÁCIO, L.B.1; SOUZA, M.M.A. 2. 1. Universidade Federal de Goiás, Goiânia – GO, Brasil ; 2. Universidade Federal de São Carlos, São 1. Universidade de Itaúna, Itaúna - MG, Brasil; 2. Carlos - SP, Brasil. Universidade Federal de Minas Gerais, Belo Horizonte - MG, Brasil. Introduction: The hematophagous mosquitoes Aedes aegypti is the main transmitter of viral diseases of Introduction: Some factors like advances in great public health impact, such as Dengue, Zika, urbanization, technology and industrial processes, Chickungunya and urban Yellow Fever. In the absence imply an increased pollution of aquatic ecosystems. of specific resources such as vaccines and effective It’s known that residues of pharmaceuticals are drugs, the only strategy available to stop transmission ubiquitous in water at low concentrations, but it’s not is vector control, either through monitoring and so clear about their potential acute and chronic toxicity elimination of breeding grounds or through chemical to living beings who live in this habitat. Thus there is control. However, the appearance of Aedes Aegypti need for greater understanding of the ecotoxicity of resistant to current insecticides, organophosphates and current drugs in aquatic systems Objectives: This pyrethroids, has increased the demand for new research aimed to evaluated the physical and chemical chemical agents to replace them. Objective: the parameters of the waters of a subbasin located in the objective of this work was to evaluate the larvicidal Central-West of the state of Minas Gerais, at two activity of Ruthenium Complex LGMC-69 against points: an upstream point and a downstream point of a Aedes aegypti. Material and Methods: The larvicidal pharmaceutical industry effluent launch. Materials and assay with LGMC–69 was performed according to a methods: The following parameters were evaluated methodology recommended by the World Health according to Rice et al. (2012)1: pH, dissolved oxygen, Organization, where for each test 20 larvae in the third temperature, chemical oxygen demand (COD), stage were exposed for 24h at different concentrations biochemical oxygen demand (BOD), dissolved solids, (in triplicate) of LGMC-69. The complex presence of suspended solids, oils and greases, color, concentration of 1 to 100 ppm was solubilized in 0.2% turbidity, sulfate, total sulfide, total iron and dimethylsulfoxide (DMSO) in water to be tested. The phosphorus. In addition, the acute toxicity was negative control used was 0.2% DMSO and the evaluated using the test with Daphnia similis. Results positive control was Temefós at 0.060 mg mL-1.(60 and discussion: The results were compared with the ppm) The mortality larvae data was used for maximum allowed value in the Brazilian legislation determination of the lethal concentration (CL) using (CONAMA Resolution 357/2005). The parameters the Probit method with 95% confidence intervals. BOD, total sulfide, color and total iron quantification Results and Conclusion: For LGMC-69 complexe, were above the levels allowed by CONAMA. The CL50, CL90 and CL99 were 3.18 ppm; 5.56 ppm and value COD found downstream (48.4 mgO2/L) was 6.1 ppm, respectively. The complex at CL99 higher than upstream (42,8 mgO2/L), demonstrating concentration was further tested in the environment that the release of the industrial effluent contributed to (non-laboratory conditions) and the results proved its increase the amount of oxygen required for chemical effectiveness as larvicidal agent with an important oxidation. The others analyzed parameters presented residual effect,when compared to the positive control. results consistent with the values allowed by the Thus, the tests indicate that LGMC-69 is an interesting current legislation. Both collection points proved new larvicide to be explored in the fight against Aedes nontoxic front of the acute ecotoxicity test. aegypti, especially in countries where the incidence of Conclusion: Some physical chemical parameters the diseases transmitted by it has reached alarming studied were above the established limits, therefore the levels, such as Brazil. release of industrial effluents must be constantly Financial support / Acknowledgments: CAPES monitored. More studies need to be developed to evaluate more precisely the interference and toxicity of pharmaceutical residues in the aquatic environments. 1Rice EW et al. Standard Methods for the examination of water and wastewater 22ed., 2012.

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Financial support / Acknowledgments: Fundação de SNP rs1695. No statistically significant association Amparo à Pesquisa do Estado de Minas Gerais was found between the polymorphisms and Hg levels (FAPEMIG) e Universidade de Itaúna (UIT). in the high frequency subgroup: GSTP1 ( 2 = 1.38, p = 0.24), GSTM1 ( 2= 0.149, p = 0.70) and GSTT1 ( 2 = 1.67, p = 0.20. Conclusion: The results did not show a positive association between the analyzed EN 19 - LEVELS OF MERCURY AND polymorphisms and high levels of Hg, however, it was POLYMORPHISM OF GLUTATHIONE S- demonstrated that the population is exposed to Hg due TRANSFERASE, IN A RIVERINE POPULATION to its fish-rich diet, thus requiring more studies to OF THE LOWER AMAZON. verify this association. Financial support / Acknowledgments: FAPESPA COSTA, F.A.A.1,2; MENESES, H.N.M.2,3; MENESES, A.A.M.1,4,5;SAMPAIO DA SILVA, D.6,7; RODRIGUES, L.R.R.1,2,4 EN 20 - FIPRONIL IN THE WATER ALTERS BEHAVIOR IN ZEBRAFISH (DANIO RERIO) 1 Programa de Pós Graduação em Recursos Naturais da Amazônia (PPGRNA/UFOPA) Santarém - PA, BEVILAQUA, F. 1; SACHETT, A.1; CHITOLINA,R. Brasil; 2 Laboratório de Genética e Biodiversidade 1; GARBINATO, C.L.1; GASPARETO, H.M. 1; (UFOPA) Santarém - PA, Brasil; 3 Instituto de Saúde MOCELIN, R.N. 2; MARCON, M.F. 2; PIATO, A.L. Coletiva (ISCO/UFOPA) Santarém - PA, Brasil; 4 2; SIEBEL, A.M. 1. Programa de Pós Graduação Sociedade Natureza e Desenvolvimento (PPGSND/UFOPA) Santarém - PA, 1.Universidade Comunitária da Região de Chapecó, Brasil; 5 Laboratório de Inteligência Computacional Chapecó – SC, Brasil; 2. Universidade Federal do Rio (UFOPA) Santarém - PA, Brasil; 6 Centro de Grande do Sul, Porto Alegre – RS, Brasil. Formação Interdisciplinar (CFI/UFOPA) Santarém - PA, Brasil; 7 Programa de Pós Graduação Sociedade, Introduction: The presence of the pesticides in surface Ambiente e Qualidade de vida (PPGSAQ/UFOPA) water is a current reality. The increase in agricultural Santarém - PA, Brasil. activity has led to the growing use these compounds, substances with great toxicological potential. Fipronil Introduction: Mercury (Hg) is one of the most is a broad spectrum phenylpyrazole insecticide. This dangerous heavy metals due to its bioaccumulative insecticide interferes with both the insect and properties, toxicity and genotoxic potential. Currently mammalian central nervous system by selectively the mercurial exposure of the Amazonian environment binding to gamma-aminobutyric acid (GABA)-gated is mainly due to deforestation related to the use of the chloride channels and antagonizing the action of land in this region. Previous works has shown a GABA. GABA receptor inhibition disrupts the reduction in glutathione levels (GSH) in individuals chloride ion control of neuronal signaling, causing environmentally exposed to Hg. Objective: To central nervous system toxicity. Thus, the neurotoxic evaluate the association between Hg levels and effects of fipronil in vertebrates are mediated, at least polymorphisms in the Gluationa S-Transferase (GST) partly, by GABAA receptors. Objectives: Here we gene in a population of the Lower Amazon. Materials investigated the behavioral alterations induced by and Methods: Hg levels were quantified in 67 fipronil in adult zebrafish (Danio rerio) exposed to individuals from a riverine community in the city of concentrations detected in surface water. Materials and Santarém (PA). Three polymorphisms were genotyped Methods: Adult wild-type zebrafish of both sexes were in the GST gene: GSTM1 (deletion), GSTT1 (deletion) exposed for a period of 7 days to fipronil in several and GSTP1 (SNP rs1695 A>G). The Kruskal-Wallis concentrations (0.5, 5.0, 50, 100 or 150 µg/L, n=10). test was applied to evaluate the relationship between The treatment was added to the tank and changed polymorphism and Hg levels in the high frequency every day until the end of the experiment. Behavioral subgroup of fish consumption. Results: The 67 analyses were done 24h after the last day of the individuals analyzed, 10.4% (n = 7) had GSTT1 exposition. The animals were individually submitted to deletion, 38.8% (n = 26) deletion in the GSTM1 gene the novel tank test for 6 minutes. The behavioral and 23.9% (n = 16) the GG genotype of the SNP parameters were quantified with Any-Maze®. The rs1695. Regarding Hg levels, 97% (n = 65) presented statistical analyses were done with one-way levels above 10μg / L, among these 61.5% (n = 40) ANOVA/Dunnett. Results: Fish exposed to fipronil presented the normal GSTM1 gene genotype and (50 µg/L) showed a significant decrease in total 38.5% (n = 25) deletion; 89.2% (n = 58) with normal distance traveled (p<0.05 x control). At 5 µg/L GSTT1 genotype and 10.8% (n = 7) deletion; 52,3% increased the number of crossings (p<0.01) while at (N = 34) with genotype AG, 24.6% (n = 16) with 150 µg/L, there was a significant decrease in the total genotype AA and 23.1% (n = 15) with genotype GG of distance traveled (p<0.0001) and in the number of ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 70 crossings (p<0.005) as well as an increase in the time chronic non-communicable disease represent higher spent in the upper zone (p<0.0001). Conclusions: probability to had symptoms of Hg intoxication. Lack These results show that fipronil alters zebrafish of access to health services, low education level and behavior, decreasing the total distance traveled, as well income evidence the susceptibility of this community as increasing the time spent in the upper zone. More to diseases and injuries. Populations such as this study studies are necessary to determine the effects of should be a priority in public health and environmental fipronil exposition on biochemical and molecular health policies. parameters in zebrafish. Financial support / Acknowledgments: FURG and Financial support / Acknowledgments: CAPES- UFPA FAPESC / UNOCHAPECO

EN 22 - DEVELOPMENTAL, CYTOTOXIC AND EN 21 - VULNERABILITY ASSOCIATED TO GENOTOXIC EFFECTS OF GLYPHOSATE ‘SYMPTOMS SIMILAR TO THOSE OF AND IMAZETHAPYR: AN INTEGRATED MERCURY POISONING’ IN COMMUNITIES APPROACH TO THE EVALUATION OF THE OF XINGU RIVER, AMAZON BASIN ENVIRONMENTAL IMPACT OF HERBICIDE FORMULATIONS DA SILVA JÚNIOR, F.M.R. 1; OLEINSKI, R. M.1; AZEVEDO, A.E.S.2, MONROE, K. 2; DOS COSTA, G. G. 1; BRITO, L. B. 1; SANTOS, T. R. M. SANTOS, M. 1; DA SILVEIRA, T. B.1; DE 1; RODRIGUES, L. B. 1; VALADARES M. C. 1; OLIVEIRA, A.M.N. 1; SOARES, M.C.F.1; OLIVEIRA, G. A. R. 1. PEREIRA, T.S 2. 1 Faculty of Pharmacy, Federal University of Goiás 1 Universidade Federal do Rio Grande - FURG, Rio (UFG), Goiânia, Goiás, Brazil. Grande - RS, Brasil; 2 Universidade Federal do Pará - Campus Altamira, Altamira - PA, Brasil. The planting of transgenic soybeans tolerant to glyphosate has greatly enhanced the consumption of The Brazilian Amazon is known to be a region with glyphosate based-formulations in Brazilian crops. high levels of mercury (Hg) in the environment and Imazethapyr is an imidazolinone herbicide which has studies point to an association between high levels of been associated with glyphosate-based formulations to natural mercury in the mother rock and the vast control glyphosate-resistant weeds. It is widely used number of clandestine gold mines. Other studies worldwide due to its high activity at low application already report the contamination of fish in this region, rates and wide spectrum of weed control. However, it as well as high levels of Hg in biological material from has been estimated that less than 0.1% of herbicides environmentally exposed populations. On the other applied to crops reach their specific targets, leaving hand, this is one of the least developed regions of the large amounts of toxic residue free to move into planet and it is necessary to understand the different environmental compartments, including water vulnerability factors in these populations that may be resources. Therefore, we investigated the acute toxicity intoxicated by this element. The purpose of the present of individual and joint formulations of Glyphosate study was to investigate the vulnerability factors in Atanor 48 (GLY) and Imazethapyr Plus Nortox communities from Xingu River – Amazon Basin (IMZT) on zebrafish early-life stages at several levels probably exposed to Hg. It was conducted a cross- of biological organization through developmental, selection study in two cities localized in Xingu River cytotoxicity and genotoxicity markers. For that, fish and the sample contained was 268 individuals. It was embryo-larval toxicity test was performed according to collated sociodemographic questions, lifestyle, diet OECD 236 followed by the Hoechst and comet assays. habits and health conditions. The majority of the GLY and IMZT formulations (tested in the sample was female, between 30 to 59 years old, had concentration range from 5 mg/L to 100 mg/L) showed less than three years of educational level and lived in similar lethal effects with LC50-96h 72.01 mg/L and the local of study more than 240 months. There was LC50-96h 73.47 mg/L, respectively. GLY induced regular fish consumption (95.9%), principally malformations such as yolk sac and pericardial carnivorous species (80.5%). The visual problem has a oedemas and tail deformities with EC50-72h > 55 highest prevalence (43.3%) between the health mg/L. Both individual formulations caused non- problems and about the symptoms of Hg intoxication, inflated swim bladder but GLY was more toxic (EC50- memory loss (42.9%), weakness (35.1%), fatigue 120 h 23.35 mg/L) than IMZT (EC50-120 h 67.56 (34.3%), mood changes (28.7%) and concentration mg/L). Individual GLY and IMZT at concentrations difficulties (27.2%) was most reported. The female below LC50-96h (0.5, 5.0 and 50 mg/L) induced sex, age over 60, educational level below 3 years of nuclear fragmentation in a concentration-dependent study, did not had flush toilet, smoke and least one way. The mixture of formulations at 3.54 mg/L GLY + ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 71

0.0065 mg/L IMZT, considering the environmental induced no significant toxicity to A. salina and was concentration (the worst scenario) had no significant classified in Category III according to the Globally developmental effect for zebrafish model; however, it Harmonized System of Classification (GHS) criteria. was observed nuclear fragmentation in the cell based For the toxicity test with of zebrafish early-life stage, whole-organism that may be associated with cell death. AMPA proved to be slightly toxic to endpoint GLY + IMZT induced no DNA damage in zebrafish mortality (LC50-96h>100mg/L) and had no sublethal larvae cells. In conclusion, an integrated approach to effects for the organism tested. In summary, the main developmental, cytotoxicity and genotoxicity metabolite of glyphosate is not toxic for the non-target screening using zebrafish model could be used to organisms in the conditions tested. assess the impact of herbicides for human and Financial support / Acknowledgments: FAPEG, environment health. CAPES and CNPq Financial support / Acknowledgments: FINEP, FAPEG, CAPES e CNPq.

EN 24 - ANALYSIS OF NITRITE CONTENT IN EN 23 - ECOTOXICOLOGICAL EVALUATION HUMAN CONSUMPTION WATER IN THE OF AMINOMETHYLPHOSPHONIC ACID RESIDENCES OF ACADEMICS OF AN (AMPA) INSTITUTION OF HIGHER EDUCATION OF SANTARÉM-PA COSTA, G. G. 1; RODRIGUES, L. B. 1; BRITO, L. B. 1; ARAÚJO, A. R. S. 1; VALADARES M. C. 1; SANTOS, G. N. V. DOS 1; SANTOS, C. C. DOS 2; OLIVEIRA, G. A. R. 1. GUEDES, F. I. A. 3; AGUIAR, R. B. 4; SANTOS, G. N. V. DOS 5; REGO, I. C. V 6; LOPES, R. B. 7. 1. Universidade Federal de Goiás, Goiânia - GO, Brasil 1 Universidade Federal do Oeste do Pará, Santarém- PA, Brasil; 2 Instituto Esperança de Ensino Superior, The presence of glyphosate and its main metabolite Santarém- PA, Brasil; 3. Instituto Esperança de Ensino AMPA (aminomethylphosphonic acid) are generally Superior, Santarém- PA, Brasil; 4. Instituto Esperança detected together and more frequently in soil and de Ensino Superior, Santarém- PA, Brasil; 5. sediments, ditches and sewers, rivers and streams. The Universidade Estadual do Amazonas, Manaus- AM, herbicide based on glyphosate is widely used in Brasil; 6- Universidade do Oeste do Pará, Santarém- agriculture due to its effective control of weeds. It has PA, Brasil; 7-Universidade do Oeste do Pará, been estimated that less than 0.1% of herbicides Santarém- PA, Brasil. applied to crops reach their specific targets, leaving large amounts of toxic residue free to move into INTRODUCTION Nitrogen in the form of nitrite is different environmental compartments. In soil, considered a nutrient, but when a low concentration of glyphosate may undergo degradation processes and/or oxygen occurs in the water, this can cause partial adsorption. It decomposes in soil mainly by microbial nitrate reduction (denitrification), thus leading to an action. The main metabolite of glyphosate is the increase in nitrite concentrations, which may mean a aminomethylphosphonic acid (AMPA) with a half-life large Bacterial activity in the area. OBJECTIVE To ranging from 23 to 958 days in soil. The objective of verify the presence of nitrite content in water, for this study was to evaluate the acute toxicity of the human consumption, in the residences of academics of AMPA for the different non-target organisms: seeds of a Higher Education Institution in Santarém-Pa. cucumber (Cucumis sativus) and lettuce (Lactuca MATERIALS AND METHODS This is an sativa), microcrustacean Artemia salina and zebrafish experimental quantitative research. The research was (Danio rerio) early-life stages. The phytotoxicity test carried out at the Esperança Higher Education with cucumber and lettuce seeds was performed Institute, in which samples of drinking water (faucet / according to USEPA (1996) at concentrations of 0; artesian well) were collected from the residences of the 0.10; 1.0; 10; 100 and 1000 mg/L and the analyzed analysis in the chemistry laboratory of the educational parameters were the relative seed germination rate and establishment. The nitrite analysis was performed from relative root growth rate. Brine shrimp bioassay was the coloration developed in the water samples, performed according to Meyer et al. (1982) and OECD assuming that: red coloring means accentuated 202 (2004) using the concentrations of 0, 5, 10, 25, 50 presence of nitrite (from 1.0 to 2.0 mg / L - qualitative and 100 mg/L. Fish embryo–larval toxicity test was and quantitative test); pink means presence of (0.25 to carried out in accordance with OECD 236 (2013) at 0.50 mg / L) and slightly pink some traces of nitrite concentrations of 0; 5; 23; 50; and 100 mg/L. AMPA (0.05 to 0.1 mg / mL) and colorless means absence of was not phytotoxic for the tested seeds with EC50 > nitrite. In this study, total anonymity was ensured, their 1000 mg/L for both analyzed parameters. AMPA names coded sequentially, personal data was kept ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 72 confidential and results were only used for research , dieldrin and endrin3. The following organochlorine purposes. RESULTS The following research had the pesticides (α-HCH, lindane, aldrin, HCB, pp'-DDE, participation of 50 academics, who according to the op'-DDD, pp'-DDD) were found in raw and methodology explained, answered the questionnaire. pasteurized bovine milk and ultra high temperature The majority of scholars reside in the urban districts of UHT And op'-DDT) and polychlorinated biphenyls Santarém-Pa. It was observed that the type of water (PCBs). The analysis of organic raw milk and raw milk source that is most used in the residence of EIHE from São Paulo found the presence of scholars, for daily consumption, comes from artesian organophosphorus pesticides 2. Presence of pesticide wells (60%). After the chemical analysis of the 50 residues in bovine milk was evidenced in all samples water samples, it was observed that all the samples of the studies analyzed, being the organochlorines and obtained a negative result, and the nitrite content was the organophosphates the most found. The presence of not significant, obtaining the colorless complection at organochlorine pesticide residue in bovine milk the moment of the experiment. CONCLUSION With samples may indicate the persistence of these the result of the chemical experiment, it can be chemicals in the environment due to their previous use concluded that the waters used for consumption are in in agriculture or their illegal use. good condition, this refers to the absence of nitrogen Financial support / Acknowledgments: Universidade compounds in water, especially nitrite, indicating the Federal do Oeste do Pará – UFOPA non-microbial activity in this substance. Financial support / Acknowledgments: Universidade do Oeste do Pará (UFOPA) EN 26 - IMPACT OF A DRUG DISPOSAL PROGRAM IN THE PREVENTION OF POSSIBLE ENVIRONMENTAL DAMAGE EN 25 - PESTICIDE RESIDUE IN BOVINE MILK FROM CERTAIN REGIONS OF BRAZIL: A MOREIRA, J. L. 1; DUARTE, H. M. P. 1; REVIEW MAGALHÃES, G. F. 1; VIANA, G. F. S. 1

REGO, I. C. V. 1; DIAS, P. 2; SANTOS, G. N. V. 3. 1 Faculdade Guanambi, Guanambi - BA, Brasil.

1 Universidade Federal do Oeste do Pará, Santarém - Introduction: Several factors can contribute to the PA, Brasil ; 2 Universidade Federal do Oeste do Pará, overuse of medications, such as irrational use and Santarém - PA, Brasil, 3 Universidade do Estado do dispensing beyond the exact amount for treatment. Amazonas, Manaus - AM, Brasil. These leftovers are often discarded incorrectly and may cause environmental contamination. Objective: Pesticide residue and their milk processing products The objective of this study was to evaluate the may pose a risk to the health of consumers. For this knowledge of the population of a semi-arid city in reason, bovine milk has been adopted in some Bahia about the discarding of the overdue or no longer countries as an indicator to measure the persistence of being used medicines, as well as collecting medicines chemical substances in agriculture and environmental with these characteristics to give the correct pollutants 6. The objective of the research was to destination. Materials and Methods: Were included analyze, through scientific studies of different regions adults of both sexes living in the city of Guanambi- of Brazil, contamination by pesticides in bovine milk. BA, who used drugs and at the time of the interview The methodology used is a review of the literature of donated drugs that had expired or are not being used. publications regarding the chosen theme. Searches After collection, the drugs were cataloged and were carried out in the SciELO, PubMed quantified. Data were tabulated and processed using “bvsalud.org.br”electronic databases. The results the SPSS v.20 software to perform descriptive, showed that in the States of Rio Grande do Sul, São analysis of variance and chi-square statistical analysis, Paulo and Distrito Federal, the analysis of pasteurized, being accepted the level of significance of p0.05). Chi- organic and conventional milk, found the pesticides, square analysis revealed a significant difference in the clomazone, chlorpyrifos and chloromiphos and knowledge of the population about the correct way of monocrotophos (organophosphates) 4. In Mato Grosso discarding drugs according to schooling (X2=4.208, do Sul, the following organochlorine pesticides were p=0.040, n=36), demonstrating that the lower the found in raw bovine milk: aldrin, DDT, mirex, schooling, the lower the knowledge about the disposal endosulfan, chlordane, dicofol, heptachlor and of drugs. Conclusion: It can be concluded that most of dieldrin1. In Piracicaba - São Paulo, analysis of the the participants discard drugs in inappropriate places, whole milk powder identified residues of the which can lead to environmental contamination by organochlorines hexachlorobenzene (HCB), lindane, these pharmaceutical products. It is believed that drug heptachlor, aldrin, heptachlor epoxide, o, p'DDE, α- disposal programs can assist in reducing this endosulfan, endosulfan sulfate, methoxychlor, p, DDE ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 73 environmental impact by providing the correct CHED protects against the damage promoted by Al in destination for overdue or unused drugs. neurons of CA1 hippocampus region. The results Financial support / Acknowledgments: Faculdade showed the CHED has protective effect against brain Guanambi toxicity of Al. Financial support / Acknowledgments: CAPES, CNPq, FAPEG EN 27 - EUGENIA DYSENTERICA DC PROMOTES PROTECTIVE EFFECT AGAINST ALUMINIUM TOXICITY IN MICE EN 28 - BLOOD LEAD LEVELS AND CHILDREN'S INTELLECTUAL FUNCTION IN NERI, H. F. S. 1; PEIXOTO, L. F.; OLIVEIRA, T. S. LEAD GLAZED CERAMIC PRODUCTION 1; COSTA E. A. 1;DOS SANTOS, F. C. A. 1; COMMUNITY OF MARAGOGIPINHO, BAHIA: GHEDINI,P. C. 1. PRELIMINARY RESULTS

1. Universidade Federal de Goiás, Goiânia - GO, BAH, H. A. F.; GOMES JUNIOR, E. A. 1; Brasil. BANDEIRA, M. J. 1; SANTOS, N. R. 1; RODRIGUES, C. F. 1; CARVALHO, C. F. 2; Aluminium (Al) is a metal of the earth's surface and RODRIGUES, Y. J. M. 1; SANTOS, S. C. 1; ANJOS, human are naturally exposed to relatively large A. L. S. 1; ARAÚJO, C. F. 3; MENEZES FILHO, J. amounts of aluminum from food, water and air. A. 1. Exposure to high levels of Al leads to neurodegenerative disorders. Studies aiming to prevent 1. Laboratório de Toxicologia, Faculdade de Farmácia, or treat aluminum’s toxic effects showed that green tea Universidade Federal da Bahia, Salvador-BA, Brasil; extract reduces aluminum-induced brain toxicity 2. Instituto de Psicologia, Universidade Federal da through its antioxidant effects. In this way, this study Bahia, Salvador-BA, Brasil; 3. Escola Nacional de evaluated the neuroprotective effect of crude Saúde Pública, Fundação Oswaldo Cruz, Rio de hydroalchoolic extract of E. dystenterica leaves Janeiro-RJ, Brasil. (CHED), a plant from Brazilian Cerrado, in a experimental model of neurotoxicity induced by Al. Introduction: Lead (Pb) is a ubiquitous environmental Male Swiss mice (30-40 g) were randomized into six contaminant known to affect children’s intellectual groups, according treatment used (p.o.): I, (distilled development. In Maragogipinho, a famous village in water 10 ml/kg); II (AlCl3 100 mg/kg); III (quercetin the Reconcavo Baiano for its artisanal ceramic 30 mg/kg); IV, V, and VI (CHED 10, 100, and 300 production, metallic Pb is used to produce Pb oxides to mg/kg, respectively). During all time of the treatment be applied as a glazer to ceramic utensils. Objectives: (90 days), the animals received distilled water (Group Evaluate the association between Pb exposure and I) or AlCl3 100mg/Kg (Groups II, III, IV, V, VI). effect on children's cognition. Materials and methods: After the treatments, mice were euthanized, the So far, 22 school-aged children (5-13 years) from an hippocampus isolated for evaluation of catalase elementary school in the entrance of the community (CAT), superoxide dismutase (SOD) and were evaluated. Pb in settled dust was collected with acetylcholinesterase (AChE) activities, Petri dishes and blood lead (PbB) levels collected in malondialdehyde (MDA) levels and histopathological cubital vein were analyzed by electrothermal changes. The results showed that group II increased absorption spectrometry (EAS). Hemoglobin and MDA levels (nmol/mg proteins) when compared with activity of the δ-ALA dehydratase enzyme were group I (0.6±0.05; 0.3±0.02, p<0.001), and groups III, measured. Children’s intellectual function was IV, V and VI, reduced MDA when compared with II evaluated using the Progressive Matrix of Raven for (0.2±0.01; 0.2±0.04; 0.2±0.01; 0.3±0.02, p<0.001). children. Data are described as mean or median, The same result was observed for II and I groups in minimum and maximum and stratified by sex, using AChE activity (nmoles/min/mg proteins) (980.3±37.8; the statistical software SPSS version 22. Results: Pb 741.8±70.3, p<0.05), and CHED decreased AChE dust loading rates at the school (mean and range) were activity (699.1±41.12; 594.7±35.0; 587.1±24.6, 462 (168 -1766) µg/m2/30 days. These levels are p<0.05). The group II reduced the SOD activity when approximately 3-folds higher than the reference value compared with I (155±21.5; 346±14.3, p<0.05), and for Pb in dust loading set by US EPA (131 µg/m2). CHED increased the activity in all doses (358±2.1; Median (range) PbB levels for boys (2.05, 0.9-5.9 445±55.4; 375±54.8, p<0.05). CAT (nmol/mg µg/dL) and girls (1.85, 1.0-3.2 µg/dL), were not proteins) showed similar results (0.015±0.001 – I; significantly different. Mean and range of Hemoglobin 0.007±0.001 – II, p<0.001) and groups V and VI (Hb) and ALA-RE were respectively 12.8 g/dL (11.0- recovery the enzyme activity (0.011±0.001; 14.9) and 15.6% (6.4–28.2). Mean (range) Raven raw 0.14±0.0008, p<0.05). Hystological analysis showed scores were 17.8 (12-28) for girls and 12.1 (5-28), ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 74 difference was borderline (p=0.07). Conclusion: Up to Results: Tetracycline concentration before enzymatic now, no significant correlation was observed between treatment was 10 μg.mL-1 and, at the end of 24 hours, children´s Raven score and PbB levels, even though a the concentration had dropped to 2.91 μg.mL-1, negative association is suggested, possibly due to the representing 71% of drug removal. Conclusion: The low sample size. This study expects to evaluate 100 enzymatic treatment showed high efficiency, because children exposed in the Maragogipinho village and there was a 71% removal of Tetracycline. Cupuaçu can compare with about 50 non exposed children from the be added to culture medium of P. sanguineus to act as urban area of Aratuipe, Bahia. laccase inducer, allowing to achieve a high laccase Financial support / Acknowledgments: Conselho activity. Nacional de Pesquisa (CNPq); Faculdade de Farmácia, Financial support / Acknowledgments: CNPq Universidade Federal da Bahia (UFBA).

EN 30 - OXYTRACYCLINE ANTIBIOTIC EN 29 - TETRACYCLINE BIODEGRADATION REMEDIATION USING LACCASE ENZYME USING FUNGAL LACCASE ENZYME PRODUCED BY PYCNOPORUS SANGUINEUS INDUCED BY CUPUAÇU AGROINDUSTRIAL AND INDUCED BY LIGNOCELLULOSIC WASTE WASTE

GOLVEIA, J. C. S.; AVELAR, E.C.M.1.; GOLVEIA, NUNES, E. S.; AVELAR, E.C.M. 1; AMORIM, I.F. 1; S.C.J.1; SOUZA, M. K. 2; HONÓRIO, D. C. T. 1; GOLVEIA, S.C.J.1; SANTIAGO, M.F. 1. SANTIAGO, M.F. 1,2. 1 Laboratório de Enzimologia e Materiais Bioativos, 1 Faculdade de Farmácia, Universidade Federal de Faculdade de Farmácia, Universidade Federal de Goiás; 2 Programa de Pós Graduação em Tecnologias Goiás, Goiânia, Goiás, Brasil. de Processos Sustentáveis, Instituto Federal de Goiás, Goiânia, Goiás, Brasil. Introduction: Since the 1970s, effluents from sewage treatment plants (ETEs) and surface waters have been Introduction: The occurrence of antibiotics in aquatic detected with concentrations in the μg.L-1 and ng.L-1 environments has become a constant concern due to range. Antibiotics are the major threat to the the potential to cause changes in ecosystems, environment due to the risk of selection of resistant negatively impacting aquatic and terrestrial organisms, strains. Studies demonstrate the efficiency of the use of as well as exercising a probable influence on the ligninolytic enzymes produced by white rot fungi in increase of microbial resistance to antibiotics. It is the removal of drugs. In industrial and environmental known that conventional effluent treatment processes applications of laccase, a great amount of enzyme is have low drug removal efficiency, leading to surface normally required, but at low cost. Agroinstustrial water contamination. Thus, bioremediation is a viable wastes containing lignocellulosic fibers can be used as solution to this type of environmental problem, since it natural inducers of laccase. Objective: This work aims explores the metabolic diversity of microorganisms to evaluate the capacity of guariroba straw to induce using enzymes that degrades complexes chemical the production of laccase by the fungus Pycnoporus compounds to simple substances. To obtain a high sanguineus, applying that enzyme on antibiotic amount of laccase enzyme, it can be used some Oxytetracycline remediation Methods: Bioremediation lignocellulosic materials, such as that materials wasted was performed in erlenmeyers containing 10 mL of by agro-industries, rich in lignin and phenolic oxytetracycline solution (10 μg.L-1), 10 mL of sodium compounds that act as laccase inducer. Objective: This acetate buffer (50 mMol.L-1, pH 5.0) and 0.524 mL of work aims the degradation of Tetracycline using the enzyme broth (200 U). The erlenmeyers were enzyme laccase extracted from the fungus Pleurotus sp incubated at 28 ° C, rotated at 120 rpm for 48 hours and induced by Cupuaçu agroindustrial waste. and sampled throughout the treatment for further Methods: The laccase production was using the fungi quantification of the antibiotic by High Performance in the presence of agroindustrial waste (1% w/v) Liquid Cromatograph (HPLC). Results: In the absence growing in potato dextrose liquid medium (0,5% of guariroba straw, the enzymatic activity of laccase (w/v). The bioremediation was performed in reached 30.55 U.mL-1, whereas in the presence of erlenmeyers containing 10 mL of Oxytetracycline straw, this value increased to 377.23 U.mL-1, solution (10 μg.L-1), 10 mL of sodium acetate buffer representing a 12-fold increase in laccase production. (10 mmol.L-1, pH5) and enzymatic extract. The The oxytetracycline concentration before the erlenmeyers were incubated at 28 ° C with a rotation of enzymatic treatment was 10 μg.mL-1 and, at the end of 120 rpm for 24 hours and sampled throughout the 48 hours, the concentration had decreased to 1.94 treatment for further quantification of the antibiotic by μg.mL-1. Conclusion: Based on these results, it can be High Performance Liquid Cromatograph (HPLC). concluded that the guariroba straw is efficient in ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 75 inducing laccase production by the fungus Pycnoporus EN 32 - CHLORPYRIFOS DURING sanguineus. Furthermore, laccase has a high potential GESTATION: A RISK TO THE OFFSPRING? for bioremediation of the antibiotic Oxytetracycline, since it was obtained 81% of antibiotic removal after GOLART, J. S. 1; GRECHI, N. 2; BOARETO, A. C. 48 hours of enzymatic treatment. 2; ANDREOTTI, B. 2; VERGARA, F. 2; Financial support / Acknowledgments: CNPq, DALSENTER, P. R. 2. FAPEG, CAPES 1. Universidade Tecnológica Federal do Paraná - PR, Brasil; 2. Universidade Federal do Paraná - PR, Brasil. EN 31 - BIOTHERAPEUTICS IN DETOXIFICATION Introduction: Chlorpyrifos is a pesticide, member of the organophosphate class, widely used in several MATTOSO, J. L. C. countries to manage insect pests on many agricultural crops. Currently, chlorpyrifos health risks are being SUMMARY Introduction: After the Second World reevaluated due to possible adverse effects, especially War, modern man has been contaminating air, water on the central nervous system. Objectives: The aim of and soil. According to the World Health Organization this study was to investigate the possible action of this (WHO), 92% of all people on the planet live in places pesticide on the behaviors related to anxiety and where air quality is out of standards and with this 6.5 depression of offspring rats exposed during pregnancy. million die from air pollution. Environmental Material and methods: Wistar rats were treated orally pollution, coupled with lack of sanitation, inadequate with chlorpyrifos (0.01, 0.1, 1 and 10 mg/kg/day) on hygiene and infections take the lives of 1.7 million gestational days 14-20. Male offspring behavior was children under 5 years of age worldwide. In 2009, evaluated on post-natal days 21 and 70 by the elevated Brazil reached the first place in the world ranking of plus-maze test, open field test and forced swimming consumption of agrochemicals. Objectives: To search test. Results: The results demonstrated that exposure to for scientific papers that examine the efficacy of the 0.1, 1 or 10 mg/kg/day of chlorpyrifos could induce use of biotherapy, the hetero-isotherapeutic anxiogenic-like, but not depressive-like behavior at bioterapeutics, as an option to aid in the detoxification post-natal day 21, without causing fetal toxicity. This of different toxic substances in humans and animals. effect was reversed on post-natal day 70. Discussion: Method: National and international bibliographical Our data show that the Chlorpyrifos could induce an research, showing scientific works that prove the anxiogenic-like behavior at doses of 0.1, 1 and 10 effectiveness of different biotherapeutics in mg/kg/day at the end of lactation (PND21) only with detoxification. Results: The reported work showed maternal exposure (GD14-20) – when the animals positive protection and detoxification results. It is spent a shorter period of time in the open arms when important that projects with scientific methodology be compared to the control group. Interestingly, this carried out mainly in universities, due to the various anxiogenic-like behavior on PND21 in the CPF 0.1, 1 advantages presented by this type of drug in and 10 groups were not evidenced on PND70, detoxification. Conclusion: We present papers that suggesting that neuroplasticity throughout the period showed the positive action of detoxification of heavy of life can positively influence the interference caused metals such as mercury, cadmium and arsenic using by exposure to CPF during the gestational phase. Even biotherapeutic hetero-isotherapeutic drugs, raising the more, the occurrence of anxiety-like behavior in the possibility of using the same idea in the detoxification initial stage of neuronal maturation (late lactation) can of the inhabitants of large urban centers against serve as a window for the consolidation of traumatic environmental pollution. The same idea in serving the events in the future. Conclusion: We showed that dependents of licit and illicit substances. It is exposure to low doses of chlorpyrifos during the recommended that research be carried out in poison gestational period (GD14-20) can trigger a condition treatment centers with scientific methodology to of anxiety-like behavior in male Wistar rats at the end publicize this possibility with the credibility related to of the lactation period. Thus, our data suggest that the university treatment and research centers. exposure to chlorpyrifos during pregnancy can promote neurological changes in circuit required for the appropriate perceptual effects in the neurodevelopment of the male offspring. Financial support / Acknowledgments: CAPES/CNPQ

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 76

EN 33 - SIMULATION OF THE SUGARCANE EN 34 - AIRBORNE MANGANESE EXPOSURE VINASSE IMPACT AS FERTILIZER OVER THE AND ASSOCIATIONS WITH YEARS BY MATHEMATICAL MODELING NEUROBEHAVIORAL EFFECTS IN SCHOOL- AND ITS TOXIC POTENTIAL AGED CHILDREN IN SIMOES FILHO, BAHIA

CORREIA, J. E. 1; CHRISTOFOLETTI, C. A. 2; RODRIGUES, J. L. G. 1; SANTOS, N. R. 1; SOTO, M. A. 1; KIANG, C. H. 1; FONTANETTI, C. BANDEIRA, M. J. 1; ANJOS, L. S. 1; CAMPOS, L. A. 1. 1; ARAUJO, C. 2; CARVALHO, C. 3; LIMA, C. S. 3; ABREU, N. 3; MERGLER, D. 4; MENEZES-FILHO, 1. UNESP Universidade Estadual Paulista - Campus J. A. 1. Rio Claro, Rio Claro- SP, Brasil ; 2. Fundação Hermínio Ometto UNIARARAS, Araras - SP, Brasil. 1. Laboratory of Toxicology, College of Pharmacy, Federal University of Bahia Salvador, Bahia, Brazil 2. Sugarcane vinasse is a residue from the fermentation Public and Environmental Health Program, National and distillation of sugarcane juice. Because it has high School of Public Health, Oswaldo Cruz Foundation, concentrations of organic matter and potassium, this Brazil 3. Institute of Psychology, Federal University of residue has been used as fertilizer for the cultivation of Bahia, Brazil 4. Centre de recherche interdisciplinaires sugarcane for more than 30 years. However,vinasse urlebien-être, lasanté, La société et l’environnement has the highest pollutant load due to high levels of (CINBIOSE), Université du Québec à Montreal, biochemical oxygen demand (BOD), chemical oxygen Canada. demand (COD) and acid pH. The high amount of vinasse generated, 15 times greater than ethanol,can Introduction: Excessive exposure to manganese (Mn), promote physical changes in the soil favoring the mainly by inhalation, can lead to its accumulation in percolation/leaching of vinasse components with toxic the brain and induce neurotoxic effects. potential. Keeping this in mind, this study simulated Epidemiological studies have been showing that the application of intermittent vinasse for three months neuropsychological domains like cognition, motor a year over 30 years in a soil with the physical and function, memory and attention may be affected by chemical characteristics of the Rio Claro formation elevated levels of Mn in children. Other effects such as using HYDRUS 2D/3D software. The possible hyperactivity and increased aggressiveness have also concentrations that would reach the ground water at been associated with excessive Mn exposure. 20m and the water resource at a distance of Objectives: To investigate the association of Mn approximately 400m were calculated based on the biomarkers levels, measured in occipital hair, toenail modeling results. Fish were exposed to these and blood, with behavioral changes in school-aged concentrations for 96 hours. Blood was removed by children living near a ferro-manganese alloy plant. cardiac puncture for micronucleus test; 3000 Material and Methods: Samples of occipital hair, erithrocytes per fish was analysed. Gills was also toenails and blood were collected from 225 children removed and fixed to morphological analysis by (7-12 years old) enrolled in four elementary schools Marinho et al. (2014) protocol. It was observed that the with different levels of exposure to Mn, based on dust percolation of the vinasse reaches the ground water Mn deposition rates. Mn in hair (MnH), toenails level in less than 3 years, reaching a maximum (MnT), blood (MnB) and also blood lead (PbB) were concentration of approximately 42% in 30 years. The determined by graphite furnace atomic absorption leachate of the vinasse begins to reach the water spectrometry. The evaluation of child’s behavior was resource located at 400m in 16 years, reaching a performed using the Child Behavior Check List concentration of 4.8% in 30 years. The results of the (CBCL) applied to the parents. Results: Median levels modeling bring a great alert to the impact that of MnH, MnT and MnB were 0.73 µg/g (0.16–8.79), sugarcane vinasse can cause in the mid-long term, 0.84 µg/g (0.15–9.29) and 8.98 μg/L (1.51–40.43), since concentrations of 2.5%, 5% caused a statistically respectively. PbB levels were 1.2 µg/dL (0.2–15.6). significant increase in micronucleus rates and nuclei MnH and MnB were not associated with any scale of abnormalities in erythrocytes, morphological changes behavior. However, significant associations between in the gills and liver of Oreochromis niloticus fish logMnT and total CBCL raw score (β=10.17, p=0.034) exposed. The 10% dilution was lethal for all exposed were observed when adjusted for sex, age, maternal IQ fish. Although the use of vinasse as a fertilizer is and logPbB. Using the Generalized Additives Models, positive for the crop in terms of productivity, its non-linear associations between MnT levels and intensive use can generate a high impact on the externalizing behavior (p=0.035) were identified, as environment and can lead to a soil salinization process. well as with the related subscales: aggressive Financial support / Acknowledgments: FAPESP (p=0.045) and rule-breaking behavior (p=0.024). In processo 2015/26157-9 addition, associations between logMnT and scores of thought (p=0.031) and social problems (p=0.027) were ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 77 also observed. Conclusion: These results suggest that in dust. PbB levels in the majority of the children these children are under the effect of Mn chronic evaluated were lower than the attention value of 5 exposure due to the emissions from the metallurgical μg/dL, and the highest levels were observed in children plant. MnT are associated with behavioral changes, from school C which is situated in a downtown area, corroborating with growing evidence that excessive where high traffic density is observed. Conclusions: Mn exposure is related to an increase in externalizing School aged children’s Mn exposure is correlated with behaviors. Mn dust levels at their schools, which in turn depends Financial support / Acknowledgments: CAPES; on the distance from the Mn ferroalloy plant. This FAPESB work is part of a wider study (PIECES Project) that aims to investigate the association between Mn exposure from industrial source and effects on EN 35 - SCHOOL-AGED CHILDREN children’s neuropsychological functions. EXPOSURE TO AIRBORNE MANGANESE Financial support / Acknowledgments: Fundação de FROM A FERROALLOY PLANT IN SIMÕES Amparo à Pesquisa do Estado da Bahia - FAPESB/ FILHO, BRAZIL Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – CAPES. RODRIGUES, J. L. G. 1; BANDEIRA, M. J. 1; ARAÚJO, C. F. S. 2; SANTOS, N. R. 1; ANJOS, A. L. S. 1; KOIN, N. L. 1; PEREIRA, L. C. 1; EN 36 - ISOLATION AND IDENTIFICATION OF OLIVEIRA, S. S. P. 1; MERGLER, D. 3; MENEZES- CYANOPEPTIDES PRODUCED BY FILHO, J. A. 1. CYANOBACTERIA (MICROCYSTIS AERUGINOSA) 1. Universidade Federal da Bahia - BA, Brasil; 2. Escola Nacional de Saúde Pública, Fundação Osvaldo KINOSHITA, K.1; RÓLDAN, M. S.1; PINTO, E.1 Cruz - RJ, Brasil; 3. Université du Québec à Montreal, Canada. 1. Universidade de São Paulo, São Paulo - SP, Brasil.

Introduction: Although manganese (Mn) is an essential Cyanobacteria are able to produce a wide variety of element, its absorption at high rates thru other routes bioactive peptides and most of them still unknown, than GI, may induce neurotoxic effects, especially which makes cyanobacteria a highly promising group neuropsychological outcomes in children. Atmospheric in the search for molecules with pharmacological emissions from a Mn transformation plant have been properties. Microginins are non-ribosomal linear impacting the municipality of Simões Filho, Bahia for peptides containing 4 to 6 amino acids. The N-terminal decades. Recently, we reported high levels of Mn in position of these compounds is always occupied by the dust fall accumulation in several elementary schools of decanoic acid derivative 3-amino-2-hydroxy-decanoic this town. Objective: To evaluate the Mn acid (Ahda), often reported as chlorinated or environmental exposure in school-aged children from methylated. This family of peptides is described as Simões Filho, Bahia, Brazil. Material and Methods: inhibitors of angiotensin converting enzyme (ECA), Four elementary schools located at different distances turning microginins into interest candidates for the from the plant were selected: coded A, B, C and D, treatment of cardiovascular diseases. The aim of this based on their Mn levels in settled dust (1797, 5152, study was to isolate and identify microginins and an 9700 and 21038 µg Mn/m²/30 days, respectively). unknown compound of m/z 583 produced by the Occipital scalp hair, toenails and blood samples were LTPNA08 strain of Microcystin aeruginosa. collected from 225 children (7 to 12 years) enrolled at Methanolic extracts of LTPNA 08 were submitted to these schools. Mn in hair (MnH), toenails (MnT) and solid phase extraction (SPE) to obtain fractions of blood (MnB), as well as blood Pb (PbB), were different polarities. The fractions obtained were quantified by graphite furnace atomic absorption analysed by liquid chromatography/electrospray spectrometry (GF-AAS). Results: Median levels and ionization coupled to high resolution quadrupole time range of MnH, MnT and MnB were, respectively, 0.73 of flight mass tandem spectrometry µg/g (0.16–8.79), 0.84 µg/g (0.15–9.29) and 8.98 μg/L (LC/ESI/QTOF/MS/MS) monitoring the characteristic (1.51–40.43). Blood lead levels were 1.2 µg/dL (0.2– fragment ions related to the Ahda residue (m/z 128, 15.6). MnH and MnT levels varied according to 142, 162 and 196). The fraction which contains the environmental exposure reflected by the Mn dust peptides of interest was submitted to preparative loading rates at each school. A positive correlation was HPLC for further purification. The separation was observed between MnH and MnT (Spearman rho, ρ= carried out in Luna C18 (2), 250 x 10 mm, 5 μm 0.395, p<0.001) and both with Mn dust loading rate reverse phase semi-preparative column using as mobile (ρ= 0.503, p<0.001). MnB, however, was not phases: (A) water with 0.1% formic acid (B) correlated with any of other biomarkers, nor with Mn acetonitrile with 0.1% formic acid. The gradient was ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 78 linearly increased from 30 to 60% B over 20 minutes EN 38 - USE OF BIOMARKERS FOR at a flow rate of 3.5 mL/min. LC conditions were ECOLOGICAL RISK ASSESSMENT BY optimized and after re-isolation, pure compounds of XENOBIOTICS ASSOCIATED WITH m/z 756, 770 and 583 were obtained. Accuracy masses AGRICULTURAL ACTIVITIES and isotopic patterns for both precursor and product ions were used to comprehensively postulate the VIEIRA, L. P. L.; LIMA, L. B. D. 1; MORAIS, P. B. structures of the observed peptides. The provision of 2; ANDRADE, R. L. T. 3; MATOS, L. V. 3; purified materials for a complete structure LACERDA, L. P. 1; MORON, S. E. 1 characterization that further confirm the proposed structures and proceeds to assess their activities is 1 UNIVERSIDADE FEDERAL DO TOCANTINS, essential to this stage. ARAGUAÍNA - TO, BRASIL; 2 UNIVERSIDADE Financial support / Acknowledgments: FAPESP, FEDERAL DO TOCANTINS, PALMAS - TO, FUSP, CAPES BRASIL; 3 Universidade Federal do Mato Grosso, SINOP - MT, BRASIL.

EN 37 - EVALUATION OF SUBCHRONIC Fishes have often been used as bioindicators for EFFECTS OF GLYPHOSATE AT environmental impacts caused by different REGULATORY LEVELS ON ZEBRAFISH anthropogenic activities, such as agricultural activities. EARLY-LIFE STAGE The alterations in biological responses are used as biomarkers in the evaluation of animal welfare. The RODRIGUES, L. B.; COSTA, G. G. 1; objective of this study was to evaluate the ecological VALADARES, M. C. 1; OLIVEIRA, G. A. R. 1. risk of xenobiotics associated with agricultural activities using tucunaré (Cichla sp.) as a bioindicator. 1 Faculty of Pharmacy – Federal University of Goias The work was carried out in the southwest region of (UFG), Goiânia, Brazil. the state of Tocantins, in the cities of Lagoa da Confusão and Pium.Water samples and specimens of The glyphosate-based formulation Roundup® is the Cichla sp. were collected in the Javaés and Formoso most used herbicide in Brazil and worldwide. In Rivers at three points (A, B and C). The concentration agricultural fields, glyphosate is sprayed on plant of the Cd, Pb, Cu, Cr, Mn, Ni and Zn metals were foliage; however, some of the chemical could be analyzed in the water and liver of the animals. deposited directly on the soil surface or carried by the Histological biomarkers (gills and liver) were wind to neighboring soils or leach after rainfall to evaluated.The average for the Cr and Mn neighboring water bodies, leading to exposure of non- concentrations in the water at the three collection target terrestrial, aquatic organisms and humans. points exceeded the values for Class 1 rivers based on Glyphosate concentration in water can reach 1.7 mg/L the resolution of the National Council for the after direct application of the herbicide formulations. Environment – CONAMA357.The hepatic level was The National Environmental Council Resolution No. significantly higher at point C for Pb and Ni than for 357/2005 establishes the limit of 65 µg/L of Point A.In the gills, the most frequent glyphosate for surface water and the Regulation No. histopathological changes were Stage I and II (mild to 2914/2011 from the Ministry of Health establishes the moderate): epithelial detachment, hyperplasia and limit of 500 µg/L of glyphosate for potable water. This lamellar epithelium hypertrophy, congestion, partial study investigated the subchronic effects of Roundup® and total lamellar fusion, edema, inflammatory on zebrafish early-life stage at legal levels allowed in infiltrate, capillary dilatation and less frequent water. For that, fish embryo-larval toxicity test was aneurysm (Stage II) and focal necrosis (Stage III), only performed according to OECD TG 236 with extended at point C.The most frequent histological changes exposure time for 168 h. Twenty embryos were observed in the liver were Stage I and II, such as individually distributed in a 24‐ well plate filled with 2 cytoplasmic vacuolation, nuclear hypertrophy, mL of Roundup® solutions at 65, 155, 280, 500 and sinusoidal dilatation, vessel congestion, inflammatory 900 μg/L. Lethal and sublethal effects were analyzed infiltrate and pycnotic nuclei. Focal necrosis was for 168 h. Genotoxic effect was evaluated by the comet observed in animals from agricultural areas.The assay. Roundup® formulation induced no significant evaluation of these biomarkers can be inferred that mortality, malformations or DNA damage for animals from areas with intense agricultural activities zebrafish larvae under the conditions tested. Our are presenting adaptive responses. This is probably due findings indicated that subchronic exposures to to the availability and bioaccumulation of some regulatory limits of glyphosate in surface and drinking xenobiotics present in the environment, such as the water is not toxic to zebrafish early-life stage. chromium that is part of the composition of fertilizers Financial support / Acknowledgments: CAPES, and agrochemicals, and it is necessary to monitor the CNPq, FAPEG quality of the aquatic ecosystem in the region. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 79

Financial support / Acknowledgments: Conselho EN 40 - ZEBRAFISH EARLY-LIFE STAGES: A Nacional de Desenvolvimento Científico e USEFUL MODEL TO INVESTIGATE THE Tecnológico - CNPq / UNIVERSIDADE FEDERAL ENVIRONMENTAL IMPACT OF DO TOCANTINS – UFT AMOXICILLIN AND ITS OXIDIZED PRODUCTS

EN 39 - TOXICITY OF THE HERBICIDE BRITO, L.B.1; OLIVEIRA, R.2; GARCIA, L. F1; PARAQUAT IN COLOSSAMA MACROPOMUM LOBÓN, G.S.1; GIL, S.G.1; RODRIGUES L.B.1; LISBOA, C.2; FERRAZ, I.B.M.2; NUNES, N.A.2; VIEIRA, L. P. L.; LACERDA, L. P. 1; MORON, S. E. GRISOLIA, C.K.2; VALADARES, M.C.1; 1. OLIVEIRA, G.A.R.1.

1 Universidade Federal do Tocantins, Araguaína – TO, 1 Faculty of Pharmacy, Federal University of Goiás Brasil. (UFG), Brazil; 2 Biological Sciences Institute - University of Brasília (UnB), Brasília, Brazil. The herbicide Paraquat (PQ) is used for post- emergency application of weeds in different crops, as Incomplete removal of amoxicillin (AMX) during example: pineapple, cotton, rice, banana, coffee. Due wastewater treatment contributes to its presence in to its high solubility, the PQ can reach ground waters environmental compartments and consequently, through leaching of soil and water, as a result it drinking water. Efforts have been made to develop provokes adverse effects on non-target organisms, novel remediation strategies to overcome the problem including fish. Colossama macropomum (tambaqui), is of antibiotic-resistant bacteria and potential toxicity to a native fish from the Amazon basin and it has a higher non-target organisms. Therefore, the objective of this commercial value. The objective of this study was to work was to evaluate subchronic toxicity of AMX and determine the average lethal concentration CL50-96H its products of oxidation for zebrafish early-life stage of PQ in tambaquis and the genotoxicity (in sublethal through developmental and biochemical markers. The concentration) using micronucleus test (MN) and oxidation products were obtained by electrochemical nuclear anomalies (NA) and subsequent recovery for 7 oxidation (EO) was carried on tap water or Na2SO4 and 15 days. Tambaqui juveniles (WT: 20, 09 ± 3, 25 solutions. Fish embryo-larval toxicity test was g; Lt: 10, 9 ± 1, 25 cm) were sampled in aquariums of performed according to OECD 236 with 168 h (non- seventy liter (3 replicates, n = 15). For the treated AMX) and 96 h (treated AMX) of exposure. determination of CL50-96H, the following Catalase (CAT), glutathione-S-transferase (GST) and concentrations were used: control 10, 15, 20, 25 and 30 lactate dehydrogenase (LDH) activities were mg.L-1. For the evaluation of genotoxicity, sublethal determined in zebrafish larvae exposed to non-treated concentration was used: control group x 10 mg.L-1 PQ AMX after 168 h. No significant subchronic effect of for 96 hours; control group x 10mg.L-1 PQ with 96 non-treated AMX on zebrafish model was observed; hours exposition and recovery for 7 and 15 days. however, significant increase in the larvae body length Subsequently to each experiment, the blood was was observed at concentrations from 6.25 to 25 mg/L. collected through the puncture of the caudal vein, with AMX at 12.5 mg/L caused a significant inhibition of syringes containing EDTA for the preparation of blood the CAT activity while GST decreased from 6.25 mg/L extensions, they were stained with Giemsa 5% for 7 after 168 h of exposure and LDH activity was minutes and, 2000 erythrocytes per animal were significantly inhibited from 1.5 to 12.5 mg/ of AMX. analyzed for quantification of MN and NA (cells with By-products from EO consisted of tap water induced “blebbed”, “lobed”, “notched” and binucleate nucleus). no significant effect on larval development while EO The statistical analyses were performed using the using 0.1 M Na2SO4 decreased significantly survival analysis of variance followed by the Tukey test and the rate in 96 h of exposure with 78% of zebrafish larvae statistical method of Trimmed-Searman-Karber. The mortality. Besides that, after 48 h of exposure a higher CL50-96H for tambaqui was 26, 66 mg.L-1. incidence of malformations was observed in treatment Frequencies of MN and NA were observed in the using 0.1 M Na2SO4 such as cardiac edema and animals exposed during 96 hours to the PQ, there was delayed yolk sac absorption. In conclusion, AMX a significant statistical difference for MN and lobed induces effects on larval development and biochemical anomaly. After 96 hours of exposure to PQ, followed changes in some concentrations and EO using tap by 7 days of recovery, the animals continued to present water could be efficient in the remediation of these greater frequency of MN, but during the 15 recovery antibiotic because it does not generate toxic by- days there was not observed significant difference of products. MN when comparing with the control groups. Financial support / Acknowledgments: Financial support / Acknowledgments: CAPES / FAPEG, CAPES and CNPq. UNIVERSIDADE FEDERAL DO TOCANTINS ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 80

EN 41 - APPLICATION OF BIOASSAYS FOR EN 42 - OXIDATIVE STRESS LEVELS THE ECOTOXICITY ASSESSMENT OF INDUCED BY MERCURY EXPOSURE IN TEXTILE DYES MADEIRA RIVERINE JUVENILE POPULATIONS (RO/BRAZIL) OLIVEIRA, G.A.R.1; LEME, D.M.2; DE LAPUENTE, J.3; DISNER, G.R.2.; BRITO, L.B.1; CARVALHO, L. V. B.; OLIVEIRA, B. F. A. 2; CESTARI, M.M.2; OLIVEIRA, D.P.4. VEGA, C. M. R. 2; LARENTIS, A. L. 1; VIEIRA, J. A. 1; MATTOS, R. C. O. C. 1; SANTOS, D.V.S 1; 1.Faculty of Pharmacy (FF) – Federal University of COSTA-AMARAL, I. C. 1; MOURÃO, D. S. 2; Goiás (UFG), Goiânia-GO, Brazil; 2. Departament of SILVA, G. P. 2; HACON, S. S. 2. Genetics – Federal University of Paraná (UFPR), Curitiba-PR, Brazil; 3. Unit of Experimental 1. Centro de Estudos da Saúde do Trabalhador e Toxicology and Ecotoxicology (UTOX) – Barcelona Ecologia Humana (CESTEH)/ Escola Nacional de Science Park (PCB), Barcelona, Spain; 4Faculty of Saúde Pública Sergio Arouca (ENSP)/ Fundação Pharmaceutical Sciences of Ribeirão Preto (FCFRP) – Oswaldo Cruz (Fiocruz), Rio de Janeiro - RJ,Brasil; 2. University of São Paulo (USP), Ribeirão Preto-SP, Departamento de Endemias Samuel Pessoa (DENSP)/ Brazil. Escola Nacional de Saúde Pública Sérgio Arouca Textile dyeing industry is one of the main sectors that (ENSP)/ Fundação Oswaldo Cruz (Fiocruz), Rio de contributes to environmental pollution, due to the Janeiro – RJ – Brasil. generation of large amounts of wastewater loaded with dyes. Approximately 2-50% of the initial dye load Introduction: The production of reactive species is an remains in the dye bath during the fixation step of dye attribute of the human metabolism and has important on fibers and it is released in the textile effluent with biological functions, but imbalances are associated serious consequences for human health and ecological with both acute and chronic diseases, as well as the balance. In this context, this study aimed to evaluate normal aging process. This imbalance, known as acute toxicity and genotoxicity of the textile dyes Oxidative Stress, can be induced by mercury exposure Direct Black 38 (DB38), an azo dye, and Reactive (Hg), leading to health risks. Objetive: This study Blue 15 (RB15), which has copper phthalocyanine evaluated the association of oxidative stress group as chromophore, on different organisms: biomarkers with environmental Hg exposure levels in cucumber (Cucumis sativus), lettuce (Lactuca sativa), children and adolescents riverine of the Madeira River and tomato (Lycopersicon esculentum) seeds, (RO/Brazil). Materials and Methods: Blood was earthworm Eisenia foetida, microcrustacean Daphnia sampled from children and adolescents from three magna and fish [rainbow trout gonad-2 cell line (RTG- riverine local communities. Hg was determined in 2)]. The phytotoxicity test with cucumber and lettuce blood and Thiols, GST and MDA in serum, using seeds was performed according to USEPA (1996). The Inductively Coupled Plasma-Mass Spectrometry (ICP- acute toxicity test with earthworm E. foetida was MS) and spectrophotometry UV-Vis, respectively. conducted by the filter paper method. D. magna acute Results: The juvenile riverine population presented the immobilization test was carried out based on OECD highest levels of oxidative stress and Hg levels in 202. DNA damage in D. magna and RTG-2 cells was blood was from the Cuniã Extractive Reserve (GST = analyzed by Comet assay. DB38 and RB15 was not 27.2 ± 4.93 mmol/L, MDA = 1.69 ± 0.27 µmol/L, Hg- phytotoxic for the tested seeds with EC50 > 1000 B = 20.6 ± 18.0 µg/L). This population also presented mg/L for both analyzed parameters. Both dyes showed the highest frequency of fish consumption. At Cuniã, no significant effect on survival of E. foetida. DB38 the mean blood Hg levels (20.6 ± 18.0 μg/L) were 2.5 and RB15 exhibited low toxicity to D. magna and were times higher than the reference value of 8 μg/L, classified as relatively non-toxic (EC50>100 mg/L). recommended by the World Health Organization DB38 induced significant DNA damage to D. magna, (WHO) for non- exposed populations.The positive while RB15 had hormesis effect on this organism. relation between Hg-Blood and GST and MDA, With respect to RTG-2 cells, RB15 did not induce adjusted by individuals characteristics, suggest an significant genotoxicity, whereas DB38 caused direct oxidative effect. Only Thiol levels showed no (alkaline Comet assay) and oxidative (hOGG1- difference between the studied communities, indicating modified alkaline Comet assay) damages in a hormetic this is a non-sensitive biomarker in the context of this concentration responses. Therefore, our results study. This is the first study performed with emphasizes the need for biomonitoring dye levels in biomarkers of oxidative stress and exposure to Hg, via waterbodies and the use of organisms from different fish intake in the Madeira River basin. Conclusion: trophic levels for predicting the textile dye toxicity, This study demonstrates the importance of oxidative due to differences in sensitivity among species. stress biomarkers in the evaluation of environmental Financial support / Acknowledgments: FAPEG, Hg exposure, since they demonstrate initial and CNPq, Fundação Araucária reversible metabolic changes, enriching health risk ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 81 assessments. The sensitivity of these biomarkers to low reproduction, although these effects were more exposure levels allows to alert to be made before a pronounced in the exposure to MC-LR, than to lysate. disease scenario develops, with more serious effects on This was denoted, for example, by decrease in growth human health, facilitating actions to mitigate, and in all the tested concentrations of 0.1, 1 and 10 μg.L-1 minimizing the health risks of exposed populations. for MC-LR, but only at 10 μg.L-1 [D-Leu1] MC-LR Financial support / Acknowledgments: The authors for lysate. In addition, the exposure to 1 μg.L-1 MC- would like to thank all volunteers, parents, children LR caused down regulation of genes related to growth and adolescents from the Cunia, Nacional and Belmont (daf-16), fertility (spe-10) and MC detoxification (gst- communities for their collaboration in this study. 2) for MC-LR, but not for lysate. The effects observed Thanks are also due to the Laboratory of Atomic for lysate and MC-LR in the contractions of the Absorption staff at the Pontifical Catholic University pharynx could suggest shared toxic mechanisms that of Rio de Janeiro (PUC-RIO/Brazil), for performing are related to PP inhibition. The causes of higher MC- the Hg blood analyses. LR toxicity compared to the lysate could be related to a higher uptake by OATP transporters or lower degradation of pure MC-LR. EN 43 - EFFECTS CAUSED BY MICROCYSTIN- Financial support / Acknowledgments: CNPq and LR AND A CYANOBACTERIAL LYSATE CAPES. CONTAINING [D-LEU1] MC-LR ON NEMATODE CAENORHABDITIS ELEGANS EN 44 - VENLAFAXINE AND HALOPERIDOL SOPEZKI, M. 1, ESTRELLA, M. J. 1, CRUZ, L. C. 1; DO NOT INDUCE TOXIC EFFECTS ON YUNES, J. S. 2, MONSERRAT, J. M. 1, LIMA, J. V. ZEBRAFISH EARLY LIFE STAGES AT 1; ZANETTE, J. 1. ENVIRONMENTAL CONCENTRATIONS

1. Programa de Pós-Graduação em Ciências PRADO, M. R. S. 1; BRIGANTE, T. A. V. 1; Fisiológicas, Instituto de Ciências Biológicas, OLIVEIRA, D. P. 1. Universidade Federal do Rio Grande – RS, Brazil; 2. Laboratório de Cianobactérias e Ficotoxinas, Instituto 1. Universidade de São Paulo, Ribeirão Preto - SP, de Oceanografia, Universidade Federal do Rio Grande Brasil - RS, Brazil. Introduction: The use of psychotropic drugs is Microcystins (MC) are potent toxins that could be becoming more frequent every year around the world. produced by Microcystis aeruginosa cells. The MC-LR Considering the recent presence of these compounds in is the most common MC variant and its toxicity is the environment and the possible toxic effects to target often associated to its capacity to inhibit protein and non-target species, they could be classified as phosphatases (PP). The objective of this study was to emerging water contaminants. The main sources of evaluated biological effects caused by MC-LR and a these compounds are the domestic and hospital treated lysate of M. aeruginosa (RST 9501 strain) originated effluents, considering that the conventional treatment from the Patos Lagoon Estuary (RS, Brazil), in the used by sewage treatment plant is not efficient to nematode Caenorhabditis elegans. Growth, fertility remove completely most of drugs, and consequently and reproduction was evaluated in a 24-h bioassay these pollutants reach the water bodies. Venlafaxine using 24-well plates, by exposing C. elegans Bristol and haloperidol are two of this drugs that have been N2 wild type to 0.1, 1 and 10 µg.L-1 of MC-LR or found in environment, such as in surface water, dilutions of a concentrated RST 9501 lysate containing wastewater effluent and even in drinking water. 0.1, 1 and 10 µg.L-1 [D-Leu1] MC-LR in M9-buffer. Objectives: It is important to understand some possible At 24-h, animals from the group exposed to 1 μg.L-1 damages to humans and ecosystems after exposure to toxin (MC-LR and lysate) were separated for analysis these drugs through contaminated water. In that way, of pharyngeal pumping or RT-qPCR. The 24-h the main goal of this research was the evaluation of exposure to 1 μg.L-1 MC-LR, or to a lysate containing acute effects of psychotropic drugs venlafaxine and 1 μg.L-1 [D-Leu1] MC-LR, was sufficient to cause a haloperidol on the early stages of zebrafish significant decreased in the worm rhythmic development. Material and methods: The test was contractions of the pharynx. This could represent an based on OECD guideline Fish Embryo Acute important adverse effect for the organism since these Toxicity (FET) Test (OECD 236). Five doses of each movements are important for feeding. It is important to drug were tested based on environmental point out that this concentration is the highest concentrations according to the literature, varying concentration recommended by the World Health between 50 ng/L to 1000 ng/L and 0,3 ng/L to 3000 Organization (WHO) for drinking water. Negative ng/L, for venlafaxine and haloperidol, respectively. effects were also observed for growth, fertility and Eggs recently fertilized were exposed to treatments for ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 82

144 hours, then the embryos were analyzed every 24h AChE activity exposed to concentrations tested. On the and endpoints of lethality and sub-lethality were exposures of 10mg/L, 15mg/L, 20mg/L, 30mg/L and evaluated. Results: Both venlafaxine and haloperidol 40mg/L there were highly significant decrease induced to coagulation, absence of inflated swim (approximately of 28,93%, 22,47%, 44,20%, 28,72% bladder, delay in hatching and skeletal malformation, and 34,13% respectively). The PVP exposure did not but the observed results were not statistically relevant show significant difference. CONCLUSIONS: O. when compared with control. Conclusion: Considering niloticus exposed to different Ag-NPs concentrations our results, we can conclude that no venlafaxine nor shown a significant decrease on the brain haloperidol induce to toxic effects on zebrafish acetylcholinesterase activity, indicating an inhibitory development at environmental concentrations. effect of Ag-NPs towards brain acetylcholinesterase of Financial support / Acknowledgments: FAPESP this species. (2016/08042-2) Financial support / Acknowledgments: CAPES

EN 45 - EFFECT ON BRAIN EN 46 - PRELIMINARY STUDIES OF THE ACETYLCHOLINESTERASE OF SILVER NANOPARTICLE EXPOSURE ON OREOCHROMIS NILOTICUS EXPOSED TO OREOCHROMIS NILOTICUS AND DANIO SILVER NANOPARTICLES RERIO USING BRAIN ACETYLCHOLINESTERASE AS BIOMARKER MATIAS, M.V.F. 1; MARINHO, C.S. 1; LIMA, M. R.1; SOARES, E. C. S 1; MACHADO, S. S. 1; MARINHO, C. S. 1; MATIAS, M. V. F. 1; SILVA, F. ZANTA, C.L.P.S.1. F. 1; SILVA, J. M. 1; SANTOS, E. L. 2, MACHADO, S. S. 1; ZANTA, C.L.P.S.1. 1 Universidade Federal de Alagoas, Maceió - AL, Brasil. 1 Institute of Chemistry and Biotechnology, UFAL, AL, Brazil; 2 Agricultural Science Center, UFAL, AL, INTRODUCTION: Nanoparticles (NPs) exhibit Brazil. unique physicochemical properties due to their nanosize. Despite a wide usage of NPs in various INTRODUCTION: Nanoparticles (NPs) have unique applications, few studies were done on their properties that are useful in a diverse range of environmental impact and possible release to aquatic applications. However, the increase in metallic NP environment. Studies have shown that the toxicity of utilization has raised concerns about their release into silver nanoparticles (Ag-NPs) is related to its the aquatic environment and their possible interactions transformation in biological and environmental media, with the biotic and abiotic components, which are not including release of silver ions, and interaction with fully understood. Aquatic biomonitoring studies have biological macromolecules. Acetylcholinesterase been using molecular bioindicators as brain (AChE, EC 3.1.1.7) main function is terminate the Acetylcholinesterase (AChE, EC 3.1.1.7) of fish. cholinergic synapses by the hydrolysis of acetylcholine AChE can hydrolyze the neurotransmitter (ACh). AChE is used as efficient biomarker to identify acetylcholine (ACh) in cholinergic synapses. Inhibition perturbations of physiological and biochemical of AChE causes the accumulation of ACh, interfering parameters caused by pollutants. The inhibitory effect with the function of the nervous system. OBJECTIVE: AChE enzyme in the brain cause disruption of nerve The goal of this study was to evaluate the effect of function and excessive ACh accumulation. silver nanoparticle in brain extracts of fish species Nile OBJECTIVE: The goal of this study was to evaluate Tilapia (Oreochromis niloticus) and Zebrafish (Danio the effect of silver nanoparticle in brain extracts from rerio). MATERIALS AND METHODS: Silver O. niloticus (Nile tilapia). MATERIALS AND nanoparticle (AgNP) was synthesized and METHODS: The Ag-NPs (cat. no 576832, Sigma characterized according to Melo et al. (2012) Aldrich) were dispersed in water according Afifi et al., employing polyvinylpolyrridone (PVP) as stabilizer. 2016. Acute toxicity study was performed with Nine specimens of each fish species, adult zebrafish exposure of Nile tilapia fingerlings to concentrations (D.rerio) and nile tilapia fingerlings (O. niloticus) were of Ag-NPs (0, 5, 10, 15, 20, 30 and 40mg/L) and exposed to AgNP wet concentration (0, 0.1mg/mL) polyvinylpyrrolidone (PVP, 5,0mg/L), with ten fish and 0.1mg/mL PVP control. After 96 hours of per aquarium of 20L in duplicate. After 96h, fish were exposure, fish were sacrificed, brain was dissected, sacrificed and their brain tissues were collected, frozen immersed in liquid nitrogen kept at -20°C for further in liquid nitrogen and kept at -20°C for further AChE activity measurements according to Ellman et analyses. AChE activity was measured according to al. (1961). RESULTS: The formation of AgNPs was Ellman et al., 1961. RESULTS: Compared to control, confirmed spectrophotometrically at 398nm and size there were significant decrease (p < 0,05) in brain distribution ranging from 46.5 to 70 nm. Compared to ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 83 control, O. niloticus and D.rerio exposed to AgNP exposure and risk factors according the questionnaire showed a decrease in brain AChE activity around of applied. Levels of ß-HCH and pp'DDE were 15.2 and 48.6%, respectively. PVP control showed no significantly higher for individuals that lived in significant difference. CONCLUSIONS: These agricultural region and pp'DDE in individuals who preliminary results showed that brain AChE can be have worked with manipulation of pesticides, used as a biomarker for fish exposure to AgNPs. indicating a remote exposure to the DDT and HCH. In Financial support / Acknowledgments: CAPES - Brazil there are no reference values for organochlorine Coordenação de Aperfeiçoamento de Pessoal de Nível pesticides in human serum. The results provide actions Superior to prevent health risks. This work is part of the Subproject Pilot of the First National Survey of Populations Exposed to Chemical Substances EN 47 - EXPOSURE TO PERSISTENT (CGVAM/SVS/MS), USP e IAL. ORGANOCHLORINE PESTICIDES IN THE Financial support / Acknowledgments: Ministério da POPULATION OF THE METROPOLITAN Saúde (CGVAM/SVS/MS), USP e IAL. REGION OF SÃO PAULO – BRAZIL

LEMES, V. R. R. 1; KUSSUMI, T. A. 1; NAKANO, EN 48 - LEVELS OF BTEX AND METALS IN V. E. 1 ; ROCHA, S. B. 2; CARDEAL DE ATMOSPHERIC AIR AND HOUSEHOLD DUST OLIVEIRA, M. C. 1; KIMURA, I. A. 1 ; KUNO, R. 2 FROM A STEEL SLAG DEPOSIT IN VOLTA ; GOUVEIA, N. 2. REDONDA, RJ

1 Núcleo de Contaminantes Orgânicos, Instituto MOREIRA, M. F. R.; BORGES, R.M.; AZEVEDO, Adolfo Lutz, São Paulo -SP, Brasil; 2 Faculdade de S.V.; CARVALHO, L.V.B.; MOREIRA, F.R. Medicina, Universidade de São Paulo, São Paulo -SP, Brasil. Fundação Oswaldo Cruz, Rio de Janeiro – RJ, Brasil

The growths of agricultural production, Introduction: Benzene, toluene, ethylbenzene, and industrialization and urbanization in Brazil have led to xylene (BTEX) may coexist in hazardous waste human exposure to multiple chemicals. In view of deposits and this has been the most common disposal these facts, it becomes important to identify practice worldwide. Toxic effects may range from populations at risk of exposure to contaminants such as damage to renal and respiratory functions to persistent organochlorine pesticides because of its hematological effects such as leukemia and aplastic severe toxic effects for human health and the risks they anemia. All metals can be toxic and are found in air, represent to public health. São Paulo Metropolitam water, and soil. Disposal of industrial waste contribute Region (SPMR) presents various contaminated sites to increasing their levels in the environment. and has one of the highest urban concentrations in the Carcinogenicity, neurotoxicity, nephrotoxicity, world. Blood donors constitute a group of healthy damages to endocrine, reproductive, and respiratory individuals and studies have shown that adequately systems are among those major toxic effects. represent the population. The study had as objective to Objective: To determine the concentration of BTEX evaluate exposure to persistent organochlorinated and metals in the atmospheric air and household dust residues in the adult population from the SPMR. The of a condominium located next to an industrial waste biological monitoring was conducted in blood serum deposit in Volta Redonda, RJ. Materials and Methods: (N = 547) for the determination of organochlorine The atmospheric air was collected during 3 hours using pesticides of blood donors of Beneficent Association a high volume sampler with a flow rate equal to 500 L of Blood Collection (Colsan) of both sexes, between min -1. A portable air sampling pump collected the the ages of 18 and 65 years and living in SPMR. All household dust, with the flow rate calibrated to 2.0 L procedures complied with the ethical guidelines. The min -1 for metals and another with 0.2 L min -1 for identification and quantification were by gas BTEX. Air sampling was carried out in 18 points chromatography coupled to micro-electron capture during two visits, assuming the industrial waste deposit detector CG/μECD for organochlorine pesticides and as a stationary source. Results: Averages found for metabolites, totalling 8205 assays. The SPSS and Stata metals in the atmospheric air were 0.005 µg m-3 for programs were used for statistical analysis. All median Cr, 0.083 µg m-3 for Mn, 0.005 µg m-3 for Ni, 0.006 values presented were lower than limit of µg m-3 for As, 0.014µg m-3 for Cd and 0.049 µg m-3 quantification (LOQ), from 0.02 to 0.16μg/dL. The for Pb. Related to household dust, the results ranged metabolites ß-HCH and pp'DDE were most frequently from 4.97 to 38.6 µg m-3 for Cr with a median of found and with maximum levels of 0.45μg/dL and 13.68 µg m-3, 0.7 to 668.0 µg m-3 for Mn, median of 1.17μg/dL, respectively. It was investigated the 17.66 µg m-3, 0.28 to 216.3 µg m-3 for Ni, median of correlation of the variables with relation to the 5.78 µg m-3, 0.62 to 2.15 µg m-3 for As, median of ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 84

1.60 µg m-3, 0.01 to 14.23 µg m-3 for Cd, median of asmajor diseases.The mean obtained for Pb-B was 2.13 0.20 µg m-3, and 0.17 a 275.5 µg m-3 for Pb, median ± 0.80 μg dL-1, while 2.80 ± 2.61 μg L-1 and 4.35 ± of 2.94 µg m-3. As to organic compounds, only 4.41μg g-1creatinine were found for Pb-P and Pb-U, benzene and toluene were detected in air, with average respectively, as well as 24,45±8,28 UL-1for the ALAD concentrations of 3.68 µg m-3 for benzene and 3.67 µg activity. Levels of Pb-B and Pb-U were within the m-3 for toluene. Conclusion: Levels found for metals range reported as reference in the literature. However, are compatible with those found in urban area. concentrations of lead in plasma were higher than However, the accumulation of these metals in domestic reference values.Eleven individuals with inhibition of dust was high, representing a health risk to the the ALAD enzyme and two individuals with ALAD 1- population. For benzene and toluene, the levels found 2 genotypes containing the polymorphic allele were are similar to those of industrial areas. detected.Associations between Pb-B, Pb-P, Pb-U and Financial support / Acknowledgments: Fundação ALAD activity were not significant. However, a Oswaldo Cruz - PAPES significant positive correlation was found between age and Pb-B levels (r = 0.5384 and p = 0.0037). Conclusion:The evaluated population presented EN 49 - BIOMARKERS OF EXPOSURE, suggestive biomarkers profile of environmental EFFECT AND SUSCEPTIBILITY FOR LEAD exposure to lead. ININDIVIDUALS ENVIRONMENTALLY Financial support / Acknowledgments: Fundação EXPOSED TO INDUSTRIAL WASTE IN VOLTA Oswaldo Cruz, Programa de Apoio à Pesquisa GRANDE IV CONDOMINIUM, VOLTA Estratégica em Saúde (PAPES). REDONDA CITY, RJ

MOREIRA, M. F. R.; COUTINHO, F.L. 1; EN 50 - EVALUATION OF MERCURIAL AZEVEDO, S.V. 1; CARVALHO, L.V.B. 1; EXPOSURE IN CAPILLARY TISSUE OF BERGAMINI, F.P.B. 1; GOMES, R.A. 1; PERINI, J. CHILDREN AND ADOLESCENTS OF TWO 2. MUNICIPALITIES OF THE AMAZON INFLUENCED BY INDUSTRIAL ACTIVITIES 1. Fundação Oswaldo Cruz, Rio de Janeiro – RJ, Brasil; 2. Centro Universitário Estadual da Zona Oeste, JESUS, M. I.; OLIVEIRA, D. C. 1; FURTADO, E. C. Rio de Janeiro – RJ, Brasil. M. 1; JESUS, I. M. 1; LIMA, M. O. 1; SERRA,E. M. F. 1; SAGICA, F. E. S. 1; CUNHA, A. J. L. A. 2. Introduction: Contamination of air, water and soil by metals from industrial processes also affects the 1 Instituto Evandro Chagas, Ananindeua-PA; 2 general population. Lead (Pb) is a cumulative metal, Universidade Federal do Rio de Janeiro, Rio de and without any physiological function. Its effects Janeiro-RJ. mainly affect the hematopoietic, nervous, cardiovascular, renal, endocrine and immune systems. The dissemination of pollutants from waste near Objective:To assess the biomarkers of exposure, effect, industrial areas can reach varied distances through air, and susceptibility for lead in a population exposed to water, soil and sediment and reach biota, exposing industrial residues in a residential condominium of communities to contaminants. Some groups are more Volta Redonda. Materials and Methods:The study vulnerable, considering different levels of population consisted of 27 adults(16 womenand 11 susceptibility. In the vicinity of the industrial complex men). A questionnaire has requested information about of Barcarena, children and adolescents are part of these socioeconomic data and investigated potential groups, being in an age group marked by biological confounders as well as signs and symptoms consistent transformations which can be altered by external and with lead contamination. Electrothermal atomic internal risk factors, leading to changes in homeostasis, absorption spectrometry was used for the and may suffer various types of diseases, Including determination of Pb in blood (Pb-B), plasma (Pb-P) e those resulting from exposure to trace metals such as urine (Pb-U) samples.The activity of the enzyme δ- mercury (Hg), considered one of the most harmful to aminolevulinic acid dehydratase (ALAD) was human and environmental health. This study is part of measured by spectrophotometry (Shimadzu 1601-A an Environmental Monitoring Program developed by UV-Visible) and polymorphism genotyping was the Evandro Chagas Institute / SVS-MS, with Human performed in real-time PCR (Applied Biosystems). Health Assessment, aiming the evaluation the exposure Results: The population consisted of women (59.26%), to Hg based on capillary tissue. We evaluated a total of Caucasian (88.89%), age between 40-50 years 527 individuals aged between 6 and 19 years of age (29.63%), residence time in the condominium from 11 from communities of Abaetetuba (Vila do Maranhão n to 20 years (74.08%), presenting sinusitis (22.2%), = 139, Vila de Beja n = 93) and Barcarena (Industrial anaemia (14.81%) and hypothyroidism (14.81%) District n = 132, Vila do Conde n = 163), both in the ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 85

State of Pará. An epidemiological inquire was applied. PER. The initial tail length induced by 1.3 J/m2 UVC The concentration of HgTotal in capillary tissue was was 56.5 ± 8.3 µm for NER and 57.1 ± 0.9 µm for determined by cold vapor atomic absorption PER, and did not differed among the experiments spectrometry (CVAAS). The following mean values of (p=0.45). The time to repair 50% of the initial DNA mercury were found: 2.1 μg / g, 1.1 μg / g, 0.7 μg / g damage (RT50) was 37.69 h in the dark and 1.57 h and 0.6 μg / g in Vila Maranhão, Vila de Beja, Vila under visible light. Under our conditions, PER repaired Industrial and Vila do Conde respectively. The level of 24 times faster than NER. Since we observed exposure in Vila Maranhão was statistically higher differences in RT50 for base excision repair system than in other locations (p <0.05). Hg levels were between ZFL and primary hepatocytes, the kinetics for statistically higher when associated with fish NER and PER systems in primary hepatocytes will be consumption (p <0.01). The mercury values obtained also evaluated. Our results also suggest the RT50 for in this study are in accordance with the Limit of NER and PER is a promising parameter to evaluate the Normality levels (LN = 2μg / g) established by the effect of contaminants on the DNA repair capacity. World Health Organization (WHO). However, Financial support / Acknowledgments: The authors although Vila Maranhão is considered a control area, thank to the Coordenação de Aperfeiçoamento de the average levels of Hg found are above the reference Pessoal de Nível Superior (CAPES) and the Brazilian values, corroborating previous studies in areas of Conselho Nacional de Desenvolvimento Científico e mining in the Amazon, and even in areas without Tecnológico (CNPq). anthropogenic action. Financial support / Acknowledgments: The authors gratefully acknowledge the financial support of the EN 52 - ANTIBIOTIC REMEDIATION OF Instituto Evandro Chagas. OXITETRACYCLIN BY LACCASE FROM PLEUROTUS OSTREATUS INDUCED BY AGROINDUSTRIAL WASTE AND DRIED BY EN 51 - EVALUATION OF THE DNA REPAIR SPRAY DRYER ACTIVITY IN ZEBRAFISH HEPATOCYTES MEDRADO, L.C.L1,2; SANTOS, E.C.1; COSTA, S. R. 1; HOFF, M. L. M. 1; SOUZA, M. M. HONÓRIO,D.C.T.1; GOLVEIA, J.C.S.; 1,2; SANDRINI, J. Z. 1,2. BEZERRA,G.D.3; PAULA, M.A. J.3; SANTIAGO, M.F.1,2. It is known that DNA is the target of several environmental contaminants. Therefore, many studies 1 Faculdade de Farmácia, Universidade Federal de seek to assess DNA damage as an attempt to develop Goiás, Goiânia, Goiás, Brasil; 2 Programa de Pós- environmental biomarkers. The DNA damage can be, Graduação em Tecnologias de Processos Sustentáveis, however, repaired by mechanisms, such as the Instituto Federal de Goiás, Goiânia, Goiás, Brasil; 3 nucleotide excision repair (NER). In the other hand, Campus de Ciências Exatas e Tecnológicas, the repair systems may be itself affected by Universidade Estadual de Goiás, Brasil. contaminants, which also compromise the DNA stability. In addition to NER, bulky lesions in the INTRODUCTION: The antibiotics are drugs used to DNA, such as thymine dimers caused by exposure to combat and prevent diseases caused by bacteria in both ultraviolet radiation, can be repaired by human and veterinary medicine and, consequently, the photoreactivation repair (PER). This repair system has environmental contamination by these compounds has not been reported in mammals yet and is present in fish gained relevance. These drugs have accumulated in the and other organisms. For this reason, fish is an environment, causing the death of microorganisms and interesting model to study the DNA repair systems. So the formation of other super-resistant strains. The lack far, most studies have only involved quantifying the of specific treatments for the removal of these expression of DNA repair genes. Understanding the contaminants led to the development of methodologies DNA repair activity in fish cellular models is crucial to degradation of these compounds. OBJECTIVE:This for the application of these models in ecotoxicological work aims to evaluate the degradation capacity of the and clinical studies. Thus, the aim of the study was to antibiotic oxytetracycline by the laccase enzyme analyze the kinetics of NER and PER in zebrafish extract produced by the fungus Pleurotus ostreatus hepatocytes. ZebraFish-Liver (ZF-L) cell line was using liquid extract and dried by Spray Dryer. exposed to 1.3 J/m2 UVC and the levels of DNA METHODS: Fungus cultivation was in potato broth damage were monitored over time. The DNA damage Dextrose medium added with 1% of plant material. was assessed using the comet assay and expressed as The enzymatic activity was evaluated prior to drying comet tail length (in µm). After UVC exposure, cells by spray dryer with the addition of adjuvants and were either maintained in the dark, to evaluate NER, or without the use of adjuvants for an evaluation of under a visible light source (1.5 J/s/m2), to evaluate adjuvant interference in the final result of the activity. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 86

The remediation assays were performed in 50 mL and in the second and third weeks the standard chow Erlenmeyer flasks, containing 10 mL oxytetracycline intake was smaller. In the first week all animals solution (10 μg.L-1), 10mL sodium acetate buffer consumed higher amounts of food. However, the (50mmol.L-1, pH 5.0) and a volume of crude extract amount of kilocalories derived from chocolate was corresponding to 200U of enzymatic activity. The higher in the ROFA + chocolate group than chocolate erlenmeyers were incubated at 28 ° C, rotated at 150 group in all weeks. Total caloric intake and body mass rpm for 24 hours and sampled periodically throughout gain were not different among groups. Abdominal fat the treatment for subsequent quantification of the deposition was lower in ROFA group when compared antibiotic concentration by High Performance Liquid to groups that received chocolate. Triglycerides, total Cromatograph. RESULTS: Açaí agroindustrial waste cholesterol and HDL were higher in chocolate exposed reached an activity of 3184.62 U.mL-1 on the seventh groups. There were no significant differences in day of cultivation. The Spray dryer process was better glucose and LDL levels. TBARS levels were higher in using maltodextrin as coadjuvant. In the ROFA + chocolate group and there were no significant oxytetracycline remediation, at the end of 24h, the differences in SOD and CAT activities in concentration of the antibiotic had fallen from 10,584 hypothalamus. Conclusion: These findings indicate μg.L-1 to 0.036 μg.L-1, representing a removal of that subchronic ROFA exposure increases the 99.6%. The antibiotic removal with the enzyme dried consumption and food preference for highly palatable using adjuvant maltodextrin was 70.4%. food, such as chocolate, and it may contributes to CONCLUSION: Both extracts of the enzyme laccase metabolic diseases. (liquid and dried) have great potential to be used in Financial support / Acknowledgments: UFCSPA, oxytetracycline remediation. CAPES. Financial support / Acknowledgments: CNPq.

EN 54 - ENVIRONMENTAL CONTAMINATION EN 53 - DOES AIR POLLUTION EXPOSURE BY LEAD FROM ARTISANAL POTTERY INCREASE PALATABLE FOOD INTAKE? GLAZING FACILITIES IN BAHIA, BRAZIL

DA SILVEIRA, C. G.1; DI DOMENICO, M. 2; BANDEIRA, M. J.; BAH, H. A. F. 1; COSTA, R. G. SALDIVA, P. H. N.2; RHODEN, C.R1. 1; GOMES-JUNIOR, E. A. 1; OLIVEIRA, S. S. P. 1; MENEZES-FILHO, J. A. 1. 1 Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre - RS, Brasil ; 2. Universidade de 1. Faculdade de Farmácia, Universidade Federal da São Paulo, São Paulo - SP, Brasil. Bahia, Salvador – BA.

Introduction: Residual oil fly ash (ROFA) results from Backgound: In the Recôncavo Region of the State of an incomplete oxidation of carbonaceous materials Bahia, Brazil lie along the estuary borders several common in areas where there is oil burning and it is a communities of ancient traditions, among them useful surrogate for ambient air particle matter. Air artisanal pottery facilities (olarias). Unfortunately, pollution exposure is associated to highly palatable most of them ignore the risk of the lead oxide glazing food intake and may induce health disturbs. Aim: To process used to provide shine and to waterproof the investigate the effects of subchronic ROFA exposure ceramic utensils. Objectives: To build evidence of lead on highly palatable food consumption and its effects (Pb) environmental contamination and possible on lipid profile and oxidative stress parameters in sources of Pb exposure that may impact the public hypothalamus. Material and Methods: Forty four male health of local communities. Material and Methods: Wistar rats were distributed into four groups according One of the main protein sources and income, the to exposure status and dietary intake: Control, ROFA, mangrove root crab, was used to biomonitor the tidal Chocolate, ROFA + Chocolate. The animals were Jaguaripe river. Six specimens were collected in three exposed to ROFA (50μg/μl) and saline by intranasal sampling points, along with the sediment in three instillation during 18 weeks concomitantly to palatable opportunities during the trimesters of 2016. Also, food and/or standard lab chow during the last 30 settled dust from the four elementary schools of the consecutive days. The animals were weighed once a main producer Village was sampled with disposable week. The parameters measured were food Petri dishes for approximately 30 days. Superficial soil consumption and preference, caloric intake and caloric samples were collected in three spots with varying efficiency, gain of body mass, abdominal fat distance from the ‘olarias’. Lead levels were deposition, as well as glucose and plasma lipids. determined by graphite furnace atomic absorption Oxidative stress parameters were assessed in spectrometry. Results: Mean and SD Pb levels in hypothalamus. Results: In the first two weeks, rats muscles, hepatopancreas and gills of the mangrove exposed to ROFA presented greater chocolate intake root crab were 0.012±0.007 µg.g-1, 0.066±0.106 µg.g- ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 87

1 and 0.522±0.523 µg.g-1, respectively. The average AChE as the most sensitive with all concentrations test in the river sediment was 13.84±1.25 mg.Kg-1. No inhibited (~ 25% of maximum inhibition, LOEC ≤ 0.6 difference was observed along the sampling site or µg/L). Behavioural analysis showed carbamazepine period. Pb levels in settled dust of interior effects on zebrafish total swimming time and distance environments were twice or three times higher in (LOEC ≤ 0.6 µg/L). The molecular analysis revealed a schools located nearer to the facilities (750 vs 210 dose-dependent effect of CBZ on gene expression. µg/m²/30 days). Likewise, Pb levels in soil sample in Although only 25 genes were deregulated in both one spot in the central area of the ‘olarias’ (1993 treatments compared to controls, we found that as both mg.Kg-1) were about 700 times higher than the two 0.6 and 2812 µg/L impaired expression of other sites further from the emission sources (2.6 development and neuronal-related genes, and activated mg.Kg-1) and 136 times higher than the reference genes associated with behavioural response. value for Pb in soil (14.62 mg.Kg-1). Conclusion: Our Altogether, our results indicate that environmental data shows that lead glazing process used in some the relevant concentrations of CBZ might affect fish pottery facilities are contributing enormously to Pb biochemical, genetic and behavioural traits. Thus, build-up in the environment, posing serious risk of Pb environmental risk of CBZ cannot be neglected toxicity to local villagers, especially children, and specially in a realistic scenario of constant input of consumers. domestic effluents in aquatic systems. Financial support / Acknowledgments: Fundação de Financial support / Acknowledgments: Brazilian Amparo à Pesquisa do Estado da Bahia; MCTI/CNPQ Ministry of Education and Ministry of Science and (Conselho Nacional de Desenvolvimento Científico e Technology of Brazil for the scholarship provided to Tecnológico). RO (CNPq BJT-A) - program Science without Borders; DSM for CAPES PhD scholarship. CKG for CNPq research grant 305741/2015-2. TSA for FAPDF EN 55 - LINKING THE CARBAMAZEPINE research grant. EFFECTS FROM CELLULAR TO POPULATION LEVEL: AN INTEGRATIVE APPROACH USING THE ZEBRAFISH EARLY EN 56 - MANGANESE EXPOSURE AND THE LIFE STAGES ONSET OF EARLY PUBERTY IN SCHOOL- AGED CHILDREN LIVING NEAR A FERRO- MURIEL LOPES DA SILVA1; ROLANDO ANDRÉ MANGANESE ALLOY PLANT: PRELIMINARY RIOS VILLACIS1; DIEGO SOUSA-MOURA1; RESULTS THAYRES SOUSA ANDRADE1; INÊS DOMIGUES2; CAROLINA ALMEIDA LISBOA1; SANTOS, N. R.; RODRIGUES, J. L. G. 1,2; NÍCHOLLAS SERAFIM CAMARGO3; LUÍS BANDEIRA, M. J. 1,2; ANJOS, A. L. S. 1, BAH, H. ALEXANDRE MUEHLMANN3; ALINE PIC A. F. 1,2; OLIVEIRA, S. S. P. 1; ARAÚJO, C. F. 3; TAYLOR4; RHAUL OLIVEIRA1 AND CESAR MENEZES-FILHO, J. A. 1,2. KOPPE GRISOLIA1. 1 Laboratório de Toxicologia, Faculdade de Farmácia, Carbamazepine is the most commonly used drug for Universidade Federal da Bahia, Salvador, Bahia, epilepsy treatment, regularly consumed by patients and Brazil; 2 Programa de Pós-Graduação em Farmácia, in constant daily doses. Due to physical chemical Faculdade de Farmácia, Universidade Federal da proprieties such high dissipation time in water, low Bahia, Salvador, Bahia, Brazil; 3 Escola Nacional de degradation rate and moderate affinity to the sediment Saúde Pública, Fundação Oswaldo Cruz, Rio de the CBZ removing rate in STP is usually low (below Janeiro, Rio de Janeiro, Brazil. 10%). Thus, CBZ and its metabolites are commonly detected and persistent pharmaceutical in the Background: Manganese (Mn) is an essential element environment. The present study aimed to investigate important for physiological processes but its the effects of CBZ on fish early life stages by means of overexposure may impact central nervous and an integrative approach linking endpoints from reproduction systems. It has been suggested that Mn molecular to population level. Validated methods were may induce precocious arising of sexual characteristics used including: (i) development; (ii) locomotor and elevation of puberty-related hormones. However, activity; (iii) biochemical markers (lactate there is a lack of evidence demonstrating it in children dehydrogenase, glutathione-S-transferase, environmentally exposed to Mn. Objective: To acetylcholinesterase and catalase) and (iv) investigate the possible association between Mn transcriptome analysis using microarray. Mortality and overexposure and onset of early puberty in school- developmental abnormalities were only found in aged children living in Simões Filho, Bahia. Methods: concentrations > 100 mg/L of CBZ. A general Dust Mn loading rates were assessed and four schools decrease in biochemical markers was observed with with very distance from the industrial plant were ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 88 selected: coded A, B, C and D with 1787, 5152, 9700 chemicals consumption and process time. Objective: and 21038 µg Mn/m2/30days, respectively. Children The purpose of this study was to evaluate and optimize aged 7 to 12 years were included. Levels of Mn in the degradation of DR343 dye by Fenton and Fenton- blood (MnB), hair (MnH) and toenails (MnTn) and Like AOP, evaluating the toxicity of the treated blood lead (PbB) were determined by graphite furnace samples. Material and Methods: BBD was applied to atomic absorption spectrometry. Tanner sexual staging Fenton and Fenton-like AOP aiming DR343 and anthropometric measurements were carried out. degradation. Iron and H2O2 concentrations, pH, and Results: A total of 225 children were evaluated, 114 process time were used as independent factors in the (50.7%) girls and 111 boys. Median and range of BBD, while dye removal and residual H2O2 were used MnH, MnTn and MnB were 0.74 µg/g (0.16–8.79), as dependent factors. The original dye ecotoxicity was 1.43 µg/g (0.15–13.30) and 9.16 µg/L (1.51–40.43), evaluated and statistically modeled on different respectively. PbB median was 1.15 µg/dL (0.3–15.6). organisms (lettuce seed, Artemia salina, and zebrafish Few children presented low stature (5%), 2.6% early-life stages). All the treated samples were thinness, 11.5% obesity and 12% overweight. Breast, submitted to the same protocols. Results: The dye genitalia and pubic hair development were positively removal was in the range from 5.8% to 100%. This correlated both with weight/age and body mass response was only affected by iron concentration in index/age z-scores. In girls younger than 8 years, 6.6% both Fenton and Fenton-like processes. The lowest presented telarche (breast development) and 4.1% levels of the H2O2 concentration and process time pubic hair while in boys younger than 9 years, 13% were effective in promotion of dye degradation. No presented pubic hair and 24.6% genital development. significant phytotoxic effect was observed in original Majority of girls were classified at M1 stage of breast dye solutions, neither after the treatments in both development and at P2 stage of pubic hair, and boys, at Fenton systems. In turn, the process time had G1 stage of genital growth and at P1 stage of pubic significant effect on A. salina mortality for the samples hair. Conclusion: No significant correlation was submitted to the Fenton process, which suggests the observed between Mn biomarkers and the age of formation of toxic oxidation by-products at the sexual secondary characteristics appearance. Puberty- beginning of the process followed by total related hormones (Prolactin, Luteinizing Hormone, mineralization of the samples. The samples treated Estradiol, Testosterone) are now being evaluated to according to desirability function did not induce lethal investigate the possible influence of Mn overexposure or sublethal effects to zebrafish early-life stage. on their serum levels. Conclusion: The studied processes can be successfully Financial support / Acknowledgments: Fundação de used to degradate DR343 dye from water, but the Amparo à Pesquisa do Estado da Bahia; MCTI/CNPQ effects of the different process factors on toxicity (Conselho Nacional de Desenvolvimento Científico e should be analyzed and optimized. Tecnológico). Financial support / Acknowledgments: Acknowledgment to IFG for granting leave for qualification EN 57 - OPTIMIZATION OF AZO DYE DEGRADATION USING FENTON AND FENTON-LIKE PROCESSES AND EN 58 - OVARIAN TOXICITY IN ROUNDUP® ECOTOXICITY EVALUATION HERBICIDE EXPOSURE IN FEMALE RATS

FERNANDES, N. C. 1; BRITO, L. B. 1; COSTA, G. CHITOLINA, R. 1; NICOLA, P. 1; SACHETT, A. 1; G. 1; OLIVEIRA, G. A. R. 1; TAVEIRA, S. F. 1; BEVILAQUA, F. 1; CUNICO, L. 1; REGINATTO, A. MARRETO, R. N. 1. 1; BERTONCELLO, K. 1; MARINS, K. 1; ZANATTA, A. P. 1; CONTERATO, G. M. M. 2; 1 Faculty of Pharmacy, Federal University of Goiás, ZANATTA, L. 1. Goiânia -GO, Brazil. 1 Universidade Comunitária da Região de Chapecó – Introduction: Dye treatment technologies as advanced Unochapecó, Chapecó - SC, Brasil. 2 – Universidade oxidation processes (AOP) have been implemented in Federal de Santa Catarina – UFSC, campus order to degrade or mineralize substances of difficult Curitibanos, Curitibanos - SC, Brasil. treatment, as azo dye Disperse Red 343 (DR343), reducing the toxicity and risks inherent to the release Introduction: Glyphosate is a non-selective herbicide of pollutants into the environment. Box-Behnken that inhibits plant growth by interfering with the Design (BBD), statistical tool used on optimization of production of essential aromatic amino acids. This processes involving experimental factors, associated to pathway is plant-specific, therefore glyphosate has the desirability function, can be used in AOP with a been considered safe for non-target organisms and view of increasing mineralization rates, reducing widely used worldwide. Although studies indicate that ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 89 glyphosate induce endocrine disruption and may of intoxication involving cattle and humans. In the adversely affect the male reproductive system, Caruaru city, Brazil, in 1996, a contamination of the evidence for its effects on ovarian function is poorly water supply led to the death of 50 hemodialysis understood so far. Objective: We evaluated the effect patients, due to CYN in the resin of the clinic's internal of exposure to glyphosate-based formulation filters. Cylindrospermopsin is a cyanotoxin produced Roundup® on plasma estradiol and progesterone levels by some cyanobacteria genus, highly soluble in water and oxidative stress parameters in rat ovaries. and bioaccumulable. Studies with tilapia (Oreochromis Materials and Methods: 40 female adult Wistar rats niloticus) have demonstrated increased oxidative stress (180-200g), aged approximately 90 days were divided related to levels of lipid peroxidation, protein and into four groups that received by oral gavage (1 mL/kg DNA oxidation as well as changes in enzymatic bw), ultrapure water (control group) or Roundup® at activity(glutathione-S-transferase, glutathione three different doses corresponding to 1.05, 10.5 and peroxidase, superoxide dismutase, catalase).The aim of 105 μg/kg of glyphosate consumed, respectively for 28 the present study was to investigate the activity of the days. Twenty-four hours later, animals were glutathione-S-transferase and catalase enzymes of the anaesthetized (10mg/kg xylazine plus 75 mg/kg biomonitor P. reticulata after acute exposure (96 hrs) ketamine, i.p.) and blood was collected. Subsequently, to CYN.Ten animals were exposed at three they were euthanized and ovarians were collected for concentrations of CYN (0.5 μg / L, 1.0 μg / L and 1.5 oxidative stress assessment. Data were analyzed by μg / L) considering concentrations from the one-way ANOVA/Tukey. Results: The highest dose of environmentally relevant established at (1.0 μg / L) glyphosate (105 μg/kg) increased plasma progesterone and a control negative group. After the exposure time, and decreased estradiol levels, which resulted in a the liver was weighed and macerated with potassium lower estradiol/progesterone ratio in comparison to phosphate buffer (0.1 M, pH 7.0) containing 0.25 M control. Additionally, 105 μg/kg of glyphosate sucrose. Protein quantification were performed by the increased the lipid peroxidation and showed a Bradford method. The activity of glutathione-s- tendency to decrease superoxide dismutase (SOD) transferase (GST) was determined by the activity in ovarian tissue, even though this decrease quantification of the reducted glutathione conjugate was not statistically significant when compared to with 1-chloro-2,4-dinitrobenzene and catalase activity control group. Conversely, the lowest doses of the was estimated by the hydrogen peroxide herbicide (1.05 and 10.5 μg/kg) did not induce changes decomposition. The results showed that the dose of 1.5 in both hormonal and oxidative parameters (lipid μg / L was able to increase the activity of GST and peroxidation, non-protein thiols and SOD activity). catalase in P. reticulata. There are reports of authors Conclusions: The results suggest that exposure to who detected an increase in enzyme activities due to Roundup® at highest concentration alters the oxidative stress. In tilapia, oxidative stress was production of reproductive steroid hormones and reported in different type of exposure, determining induces oxidative damage. Further studies should be GST activity. Thus, we conclude that acute sub-lethal performed to clarify the mechanisms involved in such exposure to CYN can promote the increased activity of effects and its implications for mammalian ovarian GST and catalase in the liver of P. reticulata. function. Financial support / Acknowledgments: Coordenação Financial support / Acknowledgments: CAPES - de Aperfeiçoamento de Pessoal de Nível Superior FAPESC, Unochapecó (PIBIC/FAPE and Artigo 170). (Capes); Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

EN 59 - ACUTE SUB-LETHAL EXPOSURE OF GUPPY (POECILIA RETICULATA) TO EN 60 - EXPOSURE TO BUPROPION AFFECTS CYLINDROSPERMOPSIN PROMOTE THE DEVELOPMENT AND BEHAVIOR OF CHANGES IN ENZYMATIC ACTIVITY OF ZEBRAFISH EMBRYOS CATALASE AND GLUTATHIONE-S- TRANSFERASE FRANCO-BERNARDES, M. F. 1; DE OLIVEIRA, R. 2; BRYAN, I. 1; LOPES, M. 1; SOUSA-MOURA1, D. SILVA, R. A. 1; RABELO, J. C.S. 1; HANUSCH, A. 1; ANDRADE, T. S. 1, GRISOLIA, C. K. 1. L. 1; SALVINO, L. G. 1; MESQUITA, L. A. 1; SABÓIA-MORAIS, S. M. T. 1. 1. Universidade de Brasília, Brasília - DF, Brasil; 2. Universidade de Campinas, Limeira - SP, Brasil. 1 Universidade Federal de Goiás, Goiânia - Goiás, Brasil. Introduction: Pharmaceuticals are the most relevant emerging contaminants that affect the aquatic Cryospermopsin (CYN) researchs have increased over ecosystems due to their universal use and to time because of their relationship to various outbreaks incomplete elimination after depuration of wastewater. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 90

Bupropion (BPP) is an antidepressant that acts via EN 61 - MERCURY LEVELS AND ITS inhibition of norepinephrine and dopamine reuptake. It RELATIONSHIP WITH GENE SOD1 IN has been found in various water resources in RESIDENTS OF THE URBAN AREA AND concentrations up to 4300 ng/L, and thus its effects on VÁRZEA OF A MUNICIPALITY OF THE LOW aquatic organisms must be studied. Objective: The AMAZONAS present study aimed to evaluate the acute toxic effects of BPP on zebrafish (Danio rerio) early life stages PEREIRA, R. J. B.; MENESES H.N.M.1,2; comprising different levels of organization including MENESES A.A.M 3,4,5; SAMPAIO DA SILVA developmental, biochemical and behavior. Material D.6,7; RODRIGUES L.R.R2,3,4. and Methods: To assess lethal and sublethal effects of BPP on zebrafish embryos, organisms were exposed 1 Instituto de Saúde Coletiva (ISCO/UFOPA); during 168 h to concentrations ranging from 0 to 150 2Laboratório de Genética e Biodiversidade, mg/L following the OECD testing guideline n. 236. Universidade Federal do Oeste do Pará, Pará - PA, The developmental endpoints evaluated included Brasil; 3Programa de Pós Graduação em Recursos hatching, edemas, malformations and loss of Naturais da Amazônia (PPGRNA/UFOPA); equilibrium. A sub-teratogenic range of concentrations 4Programa de Pós Graduação Sociedade Natureza e (from 0 to 50000 ug/L) was used to assess effects at Desenvolvimento (PPGSND/UFOPA); 5Laboratório biochemical level by measuring the activity of de Inteligência Computacional (UFOPA); 6Centro de acetylcholinesterase and behavior by analyzing Formação Interdisciplinar (CFI/UFOPA); 7Programa embryos locomotor activity. Embryonic movement de Pós Graduação Sociedade, Ambiente e Qualidade was evaluated using the automated video tracking de vida (PPGSAQ/UFOPA). system Zebrabox. Results: BPP had low acute toxicity to zebrafish embryos with a calculated168 h-LC50 of Introduction: Mercury (Hg) is a ubiquitously heavy 111.8 mg/L. However, concentrations equal or above metal present in the biosphere. Methylmercury 22.4 mg/L elicited several developmental effects such (MeHg) is the most dangerous form capable of causing as hatching delay, loss of equilibrium and tail oxidative stress in the body through free radicals such deformities. Acetylcholinesterase activity seems to do as the superoxide anion. The enzyme superoxide not be affected by the tested BPP concentrations. dismutase (SOD) is the first antioxidant defense Behavioral analysis showed that BPP affects zebrafish system to eliminate gents such as Hg. Objective: To locomotor behavior by increasing activity at low evaluate the relationship between the SOD2 gene concentrations and decreasing at higher concentrations. rs4880 polymorphism and Hg level of the urban and Organisms exposed to 8.8 and 158 μg/L showed a riverine population in Santarém, Pará. Methodology. It significant increase in locomotor activity. On the other was studied 145 individuals (73 urban area and 72 hand, embryos exposed to 50000 ug/L presented a riverine area). Hg was quantified using the atomic decrease in the total swimming distance. Conclusion: absorption spectrophotometry technique and the The locomotor assay showed to be extremely sensitive, genotyping of the rs4880 C> T polymorphism was detecting effects at environmental relevant performed by High Resolution Melting technique concentrations of BPP (8.8 μg/L). Thus, our results (HRM). Kruskal-Wallis test was applied to verify the indicate a potential risk of BPP to non-target effect of SOD2 gene polymorphism in relation to Hg organisms. level. Results. Of the 145 individuals analyzed, 42.3% Financial support / Acknowledgments: Brazilian presented the wild CC genotype, 37.7% the CT Ministry of Education and Ministry of Science and genotype and 20% the mutant TT genotype. Regarding Technology of Brazil for the scholarship provided to Hg level, CC genotype was the most frequent (19%) Rhaul de Oliveira (CNPq BJT-A/PNPD CAPES); among individuals with levels up to 10μg/L, and the Diego Sousa-Moura for CAPES PhD scholarship. genotype CT (26.3%) among individuals with levels Cesar Koppe Grisolia for CNPq research grant above 10μg /L. Regarding fish consumption, the CT 305741/2015-2; Thayres S Andrade (1250/2016) for genotype (32.1%) was the most common in the high FAPDF research grant. frequency subgroup and in the low frequency subgroup was the CC genotype (14.6%). There was no statistically significant association between increased Hg levels and rs4880 polymorphism in the population studied ( 2 = 0.10, p = 0.75). Conclusion. It was not possible to associate Hg level with the rs4880 polymorphism and more studies are needed to determine which genes are influencing the high levels of Hg found in this population. Financial support / Acknowledgments: FAPESPA

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EN 62 - QUALITY AND WATER TOXICITY IN EN 63 - GENOTOXIC AND CYTOTOXIC A SECTION OF RIVER BOIS, GOIÁS EVALUATION OF THE HYDROALCOHOLIC EXTRACT OF PORTULACA OLERACEA SOARES, S. S.; SCALIZE, P. S. 1; OLIVEIRA, G. A. R. 1; COSTA, G. G. 1; BRITO, L. 1; ARRUDA, P. N. RODRIGUES, T. N. R. 1; ANDRADE, M. F. 1; 1. PEREIRA, A. C. F. 1; VIANA, G. F. S. 1.

1 Universidade Federal de Goiás, Goiânia – GO, Brasil 1 Faculdade Guanambi, Guanambi – BA, Brasil.

The integrated management of the river basin together Introduction: Portulaca oleracea is a plant widely used with the use of bioindicators, are characterized as in the world for therapeutic and food purposes, but efficient methods in the search for the preservation of studies evaluating its safety are still scarce. Objective: aquatic ecosystems and water quality. Thus, the The present work aimed to evaluate the genotoxic and objective of the present study is to evaluate the water cytotoxic potential of the hydroalcoholic extract of P. quality in a section of the Bois River using physical- oleracea. Materials and Methods: Dehydrated and chemical and microbiological analyzes together with pulverized P. oleracea samples were submitted to a acute toxicity tests. A collection was carried out in decoction procedure for the production of the February of 2017 in 5 points distributed in a stretch of hydroalcoholic extract. The extract was rotoevaporated 49.3 km of extension, among the 10 existing and oven dried to remove all solvent. For analysis of monitoring points. The samples were collected and genotoxicity and cytotoxicity, the Allium cepa assay preserved according to the National Guide of was used. The roots of this plant were exposed to Collection and Preservation of Specimens of CETESB. concentrations of 100 and 150mg/L of the P. oleracea The analysis of water quality parameters were extract for 72h, using copper sulfate and distilled water performed according to Standard Methods (APHA; as positive (PC) and negative (NC) controls AWWA; WEF, 2012). In addition to the respectively. At the end of this period, the roots were physicochemical and microbiological analyzes, metal stained with acetic orcein and fixed on microscope analysis was performed by the Atomic Flame slide to analyze the genotoxic and cytotoxic potential Absorption Spectrometry method following USEPA by determining the mitotic index and genetic Method 3005A. The toxicity bioassay was performed abnormalities (micronucleus, binucleate cells and using the embryo-larval stage of zebrafish (Danio segmented nucleus) respectively. The results were rerio) according to OECD 236 (2013). The analysis of submitted to descriptive statistical analysis and the results did not show significant lethal effects, but ANOVA with Tukey test, being accepted as some subliminal effects that were observed after 72h, significance the value of p<0.05. Results: The mitotic such as delayed yolk sac absorption, tail deformity, index in the 150mg/L extract group (0.40 ± 0.072) was absence of pigmentation and cardiac edema. Point 9 significantly higher than NC (0.16 ± 0.018), while showed a greater effect in 72h (mean of 80%) when lower values were observed in the 100mg/L extract compared to points 2 (mean of 36.6%) and 6 (mean of group (0.24 ± 0.012) when compared to PC (0.54 ± 65%) in the same exposure period. These effects were 0.056). Higher frequency of segmented nuclei was reversed in 96h. The malformations presented with observed in A. cepa roots cells exposed to extract of greater effects in points 6 and 9 can be explained by 150mg/L (255.7 ± 26.76) when compared to NC (45.3 the high concentrations of metals such as iron, lead and ± 8.02) and 100mg/L extract group (70.0 ± 44.40). copper, which were outside the standards allowed by Significant lower difference was observed in the the current legislation. Regarding the physical, frequency of segmented nuclei in 100mg/L extract chemical and microbiological characteristics, although group when compared to PC (172.7 ± 69.64). The they did not meet the maximum allowed values of total micronucleus analysis showed a significant difference phosphorus and thermotolerant coliforms, they were only between PC and 100mg/L extract group (9.7 ± not enough to explain the effects caused in the 4.16 and 2.3 ± 0.58, respectively). No significant development of zebrafish embryos. Toxicity tests may differences were observed among groups when be representative in the assessment of public supply assessing the frequency of binucleate cells. sources, especially when releases of industrial and Conclusion: These results suggest absence of domestic effluents that degrade water quality are genotoxic and cytotoxic potential of the hydroalcoholic identified. extract of P. oleracea when evaluated by A. cepa assay. Financial support / Acknowledgments: Universidade Financial support / Acknowledgments: Faculdade Federal de Goiás. Guanambi

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EN 64 - EVALUATION OF Financial support / Acknowledgments: Doles ACETYLCHOLINESTERASE INHIBITION Indústria Farmacêutica. LEVELS IN ACEPHATE SAMPLES TREATED BY LACCASE

JARDIM, V. L. 1; SANTIAGO, M. F. 2; VALADARES, M. C. 3.

1 Faculdade de Engenharia Civil, Universidade Estadual de Campinas - UNICAMP; 2 Laboratório de Enzimologia e Materiais Bioativos - Faculdade de Farmácia, Universidade Federal de Goiás - UFG; 3 Laboratório de Farmacologia e Toxicologia Celular - Faculdade de Farmácia, Universidade Federal de Goiás - UFG.

Acephate is one of the most commonly organophosphates used in Brazil agriculture. It’s having been registered for use in 12 plant cultures. Organophosphate toxicity causes neurological damages, with several signs of cholinergic syndrome. Laccase is a multicooper enzyme produced by rot- white fungi extracellularly, that has oxidative capacity of several chemical compounds, including toxic to the environmental. The present study aims to analyze and evaluate inhibition Cholinesterase levels by toxicity of acephate achieved after treatment with laccase obtained from two strains. Pycnoporus sanguineus and Trametes villosa were grown in standard culture medium supplemented with acephate at 0,10 and 0,50 g.L-1 concentrations for 25 days. Laccase was measured every five days. The mensuration of acephate toxicity was developed from cholinesterase biochemical test by pharmaceutical industry Doles ®. The components of the analysis samples were: women blood plasma with no history of organophosphate contamination and acephate solution submitted and not to the laccase treatment. Samples were prepared by serial dilution (12.5 to 75%) and the analyzes performed in triplicate under normal environmental conditions. The acetylcholinesterase inhibition assay showed acephate toxicity reduction in all samples treated by laccase from both strains. Cholinesterase activity increased up to 91.7% compared to not treated samples. Results reached the method reference range, showing that the organophosphate acephate toxicity was within acceptable values. The assay further suggests that there was no formation of the methamidophos metabolite. The treatment of the organophosphate acephate by laccase from Pycnoporus sanguineus and Trametes villosa was effective in reducing the inhibitory capacity of the enzyme acetylcholinesterase. Thus, the use of cholinesterase activity to evaluate the microbiological treatment of acephate was shown to be a promising analytical tool not only for this specific purpose but also for uses in environmental monitoring.

EXPERIMENTAL TOXICOLOGY ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 94

EX 01 - FLORAL RESIN FROM CLUSIA ROSEA EX 02 - DEVELOPMENTAL TOXICITY OF THE INDUCED CELL DEATH IN VITRO AND DRY LEAF EXTRACT OF MOMORDICA INCREASED SURVIVAL IN BEARING TUMOR CHARANTIA LINN (CUCURBITACEAE) IN MICE RATS

SANTOS, A. P. 1; CORTEZ, A. P. 2; OLIVEIRA, C. TRAUTENMULLER, A. L.; SOARES, J. A. 1; M. A. 3; VALADARES, M. C. 4. PAULA, J. A. M. 2,3; AMARAL, V. C. S. 1,3

1 Universidade Federal de Goiás, Goiânia - GO, 1. Laboratory of Pharmacology and Toxicology of Brasil; 2 Universidade Federal de Goiás, Goiânia - Natural and Synthetic Products, Exact and GO, Brasil; 3 Universidade Federal de Goiás, Goiânia Technological Sciences Campus, State University of - GO, Brasil; 4 Universidade Federal de Goiás, Goias, Anapolis - GO, Brazil; 2. Laboratory of Goiânia - GO, Brasil. Botanical, Chemical and Biological Studies of Medicinal Plants, Exact and Technological Sciences Chronic myelogenous leukemia (CML) is a result of Campus, State University of Goias, Anapolis - GO, the constitutively activativation of the BCR-ABL Brazil; 3. State University of Goias, Graduate Program oncoprotein followed by multiple downstream in Applied Sciences to Health Products, Anapolis - signaling pathways, in special in the tyrosine-kinase GO, Brazil. pathway. A turning point for the advance in the treatment of CML was the discovery of the first Introduction: The embryonic development is very tyrosine-kinase inhibitor imatinib mesylate, sensitive to various types of chemical substances, undoubtedly revolutionised the management and especially during organogenesis. Some chemical outcome of CML patients. However, the development compounds found in plant extracts may compromise of imatinib resistance for chronic myeloid patients has the maintenance of pregnancy. Momordica charantia become a new challenge for CML treatment. Natural L. is an important medicinal plant, widely used in folk products are recognized as potential sources of medicine as anti-diabetic, anti-cancer, anti- antitumor agents. Studies show that since 1980 most of inflammatory, anti-bacterial, antifungal, anti-viral, the antitumor molecules are of natural origin. Clusia anti-helminthic, anti-oxidant, and anti-hyperlipidemic species, comprise about 250 species distributed from agent. Despite of the popular and widespread use of southern Florida to southern Brazil, and produce a this species, there is a lack of scientific knowledge large amount of polyprenilated benzophenones. The regarding its toxicity during pregnancy. Objectives: To latex and floral resin extracted from Clusia has been evaluate the possible effects of the treatment with the popularly indicated as antirheumatic agents, dry leaf extract of M. charantia L., during purgatives, digestive problems and antitumoral organogenesis, on the skeletal development of rats. activities were also observed. The purpose of this work Material and Methods: Pregnant Wistar rats were was evaluate the antileukemic potential in vitro and in randomly assigned to four groups: three treatment vivo antitumor floral resin obtained from the flowers groups and a control group. From day 6 to 15 of of Clusia rosea and elucidate the possible mechanisms pregnancy, the animals received the vehicle (control of action of this product on cell death and toxicity.The group) or the dry leaf extract at the doses of 500, 1.000 cytotoxic effect of resin of Clusia rosea was evaluated and 2.000 mg/kg. On the 21st day of gestation, in K562, HL-60 cells in vitro (MTT and Neutral Red) cesarean sections were conducted. Fetuses were and the antitumor effect in vivo in Swiss mice. Flow removed from the uterus, and examined for any cytometry, polymerase chain reaction assays and morphological abnormalities. Then, the fetuses were morphological changes using optical and fluorescence diaphanized with potassium hydroxide and stained microscopy were used to examine their ability to with Alizarin Red. The skeleton was analyzed using a induce apoptosis. C. rosea was cytotoxic to human stereomicroscope and variations and malformations leukemia cells, promoting cell growth inhibition and were recorded. Results: There were no visible gross- inducing death via apoptosis by an increase in ROS structural abnormalities and malformations in any generation, alterations in mitochondrial functions, PS fetuses from experimental groups. In addition, no externalization and activation of caspase -3, -8 ,-9 and significant differences between the groups were bax, DNA fragmentation, reducing the volume of detected in the skeletal analysis. Conclusion: The ascites tumor in animals and increased survival. Our results showed that the dry leaf extract of M. charantia results indicate that resin Clusia rosea presentes L. at the doses of 500, 1.000 and 2.000 mg/kg is not interesting cytotoxic activity against tumor cell lines teratogenic to Wistar rats. Further, the minor skeletal and antitumoral activity in ascitic ehrlich model. alterations observed in the fetuses are mostly classified Financial support / Acknowledgments: as a variation and they are reversible in the postnatal CNPQ/CAPES period.

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Financial support / Acknowledgments: PIBIC/CNPq EX 04 - ANTINEOPLASTIC PROTOTYPE DRUG (Process nº 154075/2015-9) and PBIC/UEG. LQFM018: STUDY OF THE PHARMACOKINETIC AND ALLOMETRIC PROFILE BY LC-MS/MS EX 03 - EFFECTS OF PRENATAL EXPOSURE TO THE DRY LEAF EXTRACT OF RODRIGUES, A. R.1, RODRIGUES, C. R.1, MOMORDICA CHARANTIA L. IN WISTAR GOMES, S. A.1, ZOGHAIB, A. F.1, ZOGHAIB, I. RATS V.1, OLIVEIRA, F. M.1, HONÓRIO, T. C. D.1, CUNHA, L. C.1 TRAUTENMULLER, A. L.; SOARES, J. A. 1; PAULA, J. A. M. 2,3; AMARAL, V. C. S. 1,3. 1. Núcleo de Estudos e Pesquisas Tóxico- Farmacológicas – NEPET, Faculdade de Farmácia, 1. Laboratory of Pharmacology and Toxicology of Universidade Federal de Goiás – UFG, Goiania-GO, Natural and Synthetic Products, Exact and Brasil. Technological Sciences Campus, State University of Goias, Anapolis - GO, Brazil; 2. Laboratory of Introduction: LQFM018 is a candidate to prototype Botanical, Chemical and Biological Studies of antitumor drug obtained by molecular simplification Medicinal Plants, Exact and Technological Sciences from anticancer compounds called nutlins, inhibitors Campus, State University of Goias, Anapolis - GO, of the p53-MDM2 interaction. Goal: This study aimed Brazil; 3. State University of Goias, Graduate Program to develop and validate a bioanalytical method for in Applied Sciences to Health Products, Anapolis - quantification of LQFM018 in plasma and study its GO, Brazil. pharmacokinetic. Materials and Methods: Validation contemplated the following tests: linearity, precision, Introduction: Traditionally, pregnant woman resort to accuracy, selectivity, recovery, matrix effect, residual the use of medicinal plants for believing in their safety effect and stability. The validated bioanalytical method against synthetic drugs. In this context, Momordica used the following analytical parameters: ACE® C18 charantia L. is a very popular medicinal plant typically column (100 mm × 4.6 mm, 5 µm); mobile phase: used as anti-diabetic, anti-cancer, anti-inflammatory, buffer 2 mM ammonium acetate with 0.025% formic anti-viral and anti-hyperlipidemic agent. Despite the acid and methanol (50%:50% v/v); flow: 1.2 mL/min; widespread use of this species, there is a lack of internal standard (IS): domperidone; liquid-liquid information about the risks of its use during extraction with methyl tert-butyl ether (MTBE) and pregnancy. Objectives: The aim of this study was to injection volume of 3.0 µL. LQFM018 was investigate the reproductive toxicity of the dry leaf administered to 3 females Wistar rats at 100 mg/kg, extract of M. charantia L. in Wistar rats. Material and i.p. After administration, 0.5 mL samples of blood Methods: The dry extract, at the doses of 500, 1.000 were collected by cannulation of the left jugular vein and 2.000 mg/kg, or the vehicle was given orally to with heparinized syringe. Results and Discussion: The Wistar rats from day 6 to 15 of pregnancy. Body method was linear from 10 to 15,000 ng/mL, intrarun weight gain, food and water intake were recorded daily precision was ranged from 0.6% to 5.5%, interrun throughout gestation. Cesarean sections were carried from 1.8% to 6.7%, accuracy from 99.0% to 107.0% out on the 21st day of gestation, and the total number and recovery of 74.1% ± 4.9%. Retention times were of fetuses, number of live and dead fetuses, individual 3.16 min (LQFM018) and 1.81 min (IS). fetal weight, reabsorption and implantation sites, Pharmacokinetic parameters (mean ± SD) were: t½ = placenta weight and number of corpora lutea were 2.89 h ± 2.0; ClT/F = 22.01 mL/min/kg ± 13.5; Vd/F = recorded. Results and Discussion: During 5.48 L/kg ± 3.6. To Vd value, applying the allometric organogenesis, there was a reduction in the mean scaling for an adult weighing 70 kg, a value of 1954.3 maternal body weight gain and in water intake in all L/kg is found, which suggests extravascular treatment groups when compared with the control accumulation of the compound. In allometric scaling, group (p0.05). Conclusion: The treatment with the dry the value of half-life corresponds to 12.6 h. Total leaf extract of M. charantia L. at the doses of 500, clearance value found resulting in a value of 1800.4 1.000 and 2.000 mg/kg induced maternal toxicity by mL/min for a person weighing 70 kg, indicating decreasing the maternal body weight gain and reducing greater participation of hepatic elimination the food and water intake during organogenesis. On mechanisms once the fact that the fat-soluble molecule the other hand, embryotoxicity was not observed in all of the compound requires a prior biotransformation to treated groups as well as any reproductive alterations increase its polarity so that it can be eliminated. in the dams. Conclusion: Results, extrapolated to humans, revealed Financial support / Acknowledgments: PIBIC/CNPq large half-life, high Vd and high CLT, allowing the (Process nº 154075/2015-9) and PBIC/UEG comprehension that the prototype showed good tissue distribution profile and was extensively eliminated. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 96

Financial support / Acknowledgments: CAPES; difference in the absolute and relative organ weights NEPET UFG; INSTITUTO DE CIÊNCIAS among the groups. Under light microscopy, all groups FARMACÊUTICAS-ICF. exposed to diuron presented simple hyperplasia in the kidney. Significant urinary bladder cytotoxicity was observed only under SEM in the FP groups. EX 05 - DAMAGE BY TECHNICAL OR Conclusions: TP and FP diuron are harmful to the FORMULATED DIURON ON THE urothelium and to the reproductive parameters of male UROTHELIUM AND REPRODUCTIVE Wistar rats, being the FP particularly cytotoxic to the PARAMETERS OF MALE WISTAR RATS urinary bladder urothelium. Financial support / Acknowledgments: CAPES and SALES, B. C. P. 1; FAVA, R. M. 1; NASCIMENTO TOXICAM. E PONTES, M. G. 1; SANTOS, I. P. 1; GARCIA, P. C. 2; DE CAMARGO, J. L. V. 1; OLIVEIRA, M. L. C. S. 1 EX 06 - CARBENDAZIM FORMULATIONS: THE SAME ACTIVE COMPOUND, SAME 1. Universidade Estadual Paulista (UNESP) Faculdade CONCENTRATION AND DIFFERENT de Medicina, Departamento de Patologia, Núcleo de TOXICOLOGICAL CATEGORIZATION AT Avaliação do Impacto Ambiental sobre a Saúde ANVISA Humana (TOXICAM), Botucatu - Sp, Brasil ; 2. Universidade Estadual Paulista (UNESP) Faculdade de TOLLSTADIUS, B. F. 1; VALADARES, M. C. 1. Medicina, Hemocentro de Botucatu, Botucatu - SP, Brasil. 1. FACULDADE DE FARMÁCIA – UNIVERSIDADE FEDERAL DE GOIÁS. Introduction: Wistar rats exposed during two years to 2500ppm dietary technical diuron (3-(3,4- Introduction: Carbendazim is a benzimidazole dichlorophenyl)-1,1-dimethylurea) developed high fungicide widely used in Brazil. The US incidence of urothelial tumors, being classified by the Environmental Protection Agency have classified U.S. Environmental Protection Agency as a Carbendazim as possible carcinogen in human (Class ‘‘known/likely human carcinogen’’. During the last C) with toxicological categorization of class III. It has decades our laboratory conducted studies on the no register for food uses in the US, and also is not urothelial toxicity of the diuron technical product (TP) approved in several countries on Europe. However, it provided through diet to male Wistar rats. The dietary is largely used in Brazil to plague control on cultures influence of the diuron formulated product (FP), such as: cotton, beans, citrus, soy, and others. There composed by 50% TP and 50% inert ingredients, has are a range of commercial products that has not been evaluated in rats. FP diuron was harmful to Carbendazim as active ingredient, however these reproductive system of male P. sicula lizards exposed different products formulations have different to 1100ppm during 3 weeks; the toxic effects were toxicological categorization at ANVISA. Objective: more prominent in the groups receiving FP diuron in Discuss about different formulations of Carbendazim drinking water. Objective: This study aimed to with same concentration, have shown different compare the potential toxicity of diuron TP or FP toxicological classification. Material and Methods: 1250ppm offered through diet or drinking water on the Data were obtained from AGROFIT (Brazilian urothelium and reproductive parameters of rats. database of pesticides – Sistema de Agrotóxicos Material and methods: Fifty six-week old Wistar male Fitossanitários). Results: There are 25 registered rats were allocated to five groups (control, FP diet, FP commercial products on AGROFIT that have drinking water, TP diet, TP drinking water) and Carbendazim as the active ingredient. Besides the exposed to 1250ppm diuron for 13 weeks. At the end, Carbendazim concentration be the same in all sperm parameters, and body, kidney, testis and formulations, their toxicological categorization ranges epididymis weights, and kidney and testis from I – Extremely toxic, to IV – low toxicity. The histopathology were evaluated. Urinary bladders were product labels do not show what are the inert sectioned in half, and processed for histology and ingredients, neither their concentrations. Conclusion: Scanning Electron Microscopy (SEM). Results: Diuron The composition or concentration of inert ingredients, induced changes in sperm morphology, decreased in or even the interaction of them with Carbendazim can motility, viability and in sperm concentration. All be the reason of these discrepancies of toxicity, groups exposed to diuron decreased body weights. whereas Carbendazim toxicological classification is Food consumption was lower only in the groups III, and its products have classifications from I to IV. exposed to diuron through diet. Water consumption The companies are not obligated to describe all the was lower in all diuron groups, mainly in those constituents of their products, what makes harder to exposed through drinking water. There was no know what has been used in that formulation and to ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 97 study the interaction between compounds in the than the European legislation recommendation for “mixtures of real life”. cosmetic use. Financial support / Acknowledgments: CNPq, Financial support / Acknowledgments: Processo FUNAPE. Fapesp 2016/15660-4

EX 07 - BASIC RED 51 HAIR DYE INDUCE TO EX 08 - ROLE OF iNOS IN ETHANOL-INDUCED DNA DAMAGE IN IMMORTALIZED HUMAN RENAL TOXICITY KERATINOCYTES CELLS AT LOW CONCENTARTION SILVA, C. B. P. 1; TIRAPELLI, C. R. 2

MINI, C. A. 1; OLIVEIRA, D. P. 1 1 Programa de Pós-graduação em Toxicologia, Faculdade de Ciências Farmacêuticas de Ribeirão 1. Universidade de São Paulo, Ribeirão Preto-SP, Preto, Universidade de São Paulo, Ribeirão Preto - SP, Brasil. Brasil; 2 Laboratório de Farmacologia, Escola de Enfermagem de Ribeirão Preto, Universidade de São Procedures for modify hair color are common practices Paulo, Ribeirão Preto - SP, Brasil. through the centuries. In Roman Empire, a mixture of lead with vinegar was used to disguise the gray hair. Introduction:Ethanol consumption is described to However, when L’oreal manufactured the first hair induce the expression of the inducible isoform of the coloring products, the use of hair dyes for aesthetics enzyme nitric oxide was greatly increased. Nowadays, approximately a synthase(iNOS).Objective:Evaluate the role of iNOS quarter of Brazilian population uses hair dyes. in the renal toxicity induced by chronic ethanol However, International Agency Research on Cancer consumption.Materials and Methods:Male C57BL/6 (IARC) classified some substances present in these wild type(WT) and iNOS knockout(iNOS-/-) mice(50- products as toxic, such as aromatic amines and azo 80 days old/20-25g) were randomized into 4 group. Additionally, some studies showed the groups:Control(C); Ethanol(E); iNOS(iNOS-/-) and carcinogenic potential and risk of cancer related to hair Ethanol iNOS(E-iNOS-/-) (Protocol CEUA dye exposure. Basic Red 51 is used in semi permanent 14.1.847.53.0). The animals were treated with water or and permanent hair dyes, and this compound have ethanol 20%(v/v) for 10 weeks. Urea, creatinine, K+ aromatic amines and azo group in chemical structure. and Na+ serum levels were determined to evaluate Because this, the objective was evaluate the Basic Red renal function. The renal cortex was collected for the 51 genotoxic effects in HaCaT (Keratinocytes) cells. analysis of superoxide anion(O2˙-), hydrogen For that we used HaCaT cell in monolayer culture with peroxide(H2O2), nitrate/nitrite(NOx), 80% confluence in flask. The cells were plated (2x lipoperoxidation(TBARS), catalase(CAT) and 105) and treatment time was 4 hours, they were superoxide dismutase(SOD) activities, besides Nox1, collected and homogenized in low melting point Nox2, Nox4 and SOD2 protein expression. Data were agarose at 37ºC, placed in a pregelatinized sheet and expressed as the means± standard error of the mean. covered with laminula. The cells were lysed for 24 Statistical analysis was performed by two-way hours, submitted to electrophoresis for 20 minutes at ANOVA followed by Bonferroni 25V and 300mA, neutralized with HCl and fixed with (p<0.05).Results:Long-term ethanol consumption ethanol 95º. We evaluate 100 nucleotides per increased creatinine serum levels in WT and iNOS-/- concentration using fluorescence microscope with mice (mg/dL;C:0.14±0.01; E:0.23±0.03; iNOS-/- green filter and 3.5x Gel red solution. Dunnet’s test :0.16±0.01; E-iNOS-/-:0.23±0.02;n=5-7). Ethanol- was used for statistical evaluation. The tests treated animals showed increased levels of O2˙- concentration were: 5 µg/mL; 15 µg/mL; 30 µg/mL; (RLU/mg protein;C:728.2±57.7; E:1483.7±140.1; 40 µg/mL; 50 µg/mL PBS and methyl iNOS-/-:565.1±62.3; E-iNOS-/-:1018.8±141.9 n=7-8) methanesulfonate were used as control. We detected and TBARS(nmol/mg protein;C:6.2±0.6; E:9.1±0.7; genotoxic effects significant (p<0.01) different from iNOS-/-:4.5±0.5; E-iNOS-/-:5.8±0.5;n=7-9), however the negative control at 30 µg/mL of Basic Red this increase was lower in E-iNOS-/-, when compared 51concentration. However, after 40 µg/mL of cell to WT animals. NOx levels(nmol/mg exposure, the effects seem to decrease (p<0.05). But protein;C:9.1±0.9; E:7.2±0.8; iNOS-/-:2.0±0.2; E- we attributed these results to cytotoxic effects. At 50 iNOS-/-:2.6±0.3;n=7-19) were not altered by ethanol, µg/mL, we observed an increase on cytotoxic effects but the renal cortex from iNOS-/- mice showed lower intensity and no DNA damage was not observed. levels than WT. Increased Nox4 expression was Therefore, we concluded the use of Basic Red 51 for detected in WT mice treated with ethanol. Treatment hair coloring should be carefully evaluated considering with ethanol did not affect CAT and SOD activities, that we observed DNA damage at concentrations lower Nox1, Nox2 and SOD2 protein expression or the levels ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 98 of H2O2, urea, K+ and Na+.Conclusion:Chronic Financial support / Acknowledgments: CAPES ethanol consumption decreases glomerular filtration and increases O2˙- and TBARS levels as well as Nox4 expression in cortex renal, and these responses are mediated by iNOS. These results suggest a role for EX 10 - EXPOSURE TO VALERIANA iNOS in renal toxicity induced by ethanol OFFICINALIS DURING LACTATION DID NOT consumption. CAUSE EFFECTS ON MATERNAL BEHAVIOR Financial support / Acknowledgments: FAPESP / AND CLINICAL TOXICITY CAPES CARVALHO, D. P. 1; PETERS, V. M. 1; GUERRA, M. O. 1. EX 09 - EFFECTS OF MATERNAL EXPOSURE TO ELECTROMAGNETIC RADIATION 1 Centro de Biologia da Reprodução da Universidade EMITTED BY MOBILE PHONE ON Federal de Juiz de Fora – UFJF – Juiz de Fora (MG), ENDOCRINE SYSTEM OF ADULT OFFSPRING Brasil 1. IN RATS Introduction: Psychiatric illness, as postpartum SANTOS, T. R.1; CARVALHO, D. P.2; REZENDE, depression and anxiety, are common on the perinatal C. O.2; GARCIA, R. M. G.2. period. These conditions can impair the level of functioning and ability for the woman who is lactating 1. Universidade Estadual do Rio de Janeiro, Rio de to care for herself and fetus. The treatments in these Janeiro - RJ, Brasil; 2. Universidade Federal de Juiz de conditions are very important, but the risks of Fora, Juiz de Fora - MG, Brasil. antipsychotics for mothers are still unclear due to the lack of safety data. Valeriana officinalis is used in Introduction: The number of mobile phone users is herbal medicine of many cultures as mild sedatives and growing rapidly. This increased use has caused some tranquilizers. It is believed that these effects derive concern about potential adverse effects of from interactions with GABA receptors and electromagnetic fields in the general population. Our metabolism. Clinical studies have been demonstrating group have been developing studies that showed good results on the treatment with Valerian. However, effects on reproductive parameters and a programming we did not find studies of the treatment with this drug effect on the content of adrenal catecholamine’s and in the period of lactation. Aims: Analyze the effects of serum glucose in adult offspring due the exposure to treatment with valerian during lactation on maternal mobile phones during the gestational period, which behavior and clinical toxicity. Methods: Approved by could influence the energetic metabolism of these CEUA/UFJF (Protocol number 014/2015). After the animals. Little is known about the programming effect birth of its pups Wistar rats were randomized in 5 of this type of radiation on energetic metabolism. groups (n=15) that received treatment from 2º to the Evidence has been shown that radiation is capable of 10º days of lactation by oral administration: control affecting the thyroid cells increasing apoptosis and (1ml distilled water), vehicle (1 ml distilled water, leading to hypothyroidism, which also could influence 20% glycerin) and three treated (500 mg/Kg/day, 1000 the metabolism. Aims: Our aim was to determine the mg/Kg/day, 2000 mg/Kg/day of valerian, 20% potential effects of mobile phone radiation (1800 glycerin). Considering the possibility of general toxic MHz) on the metabolism of adult offspring exposed effects or maternal behavior alterations due the drug during the intrauterine period. Methods: Approval by treatment, during all treatment period, mothers were the CEEA/UFJF (Protocol number 98/2012). Wistar daily observed regarding the following variables: female rats in the 1st day after intercourse were Clinical toxicity: Piloerection, diarrhea, randomized into two groups: exposed to radiation chromodacriorrhea, loss or weight gain, feed and water (n=10) and non-exposed to radiation (n=10). The intake; Maternal Behavior: Posture of breastfeeding, exposure took place from 1st to 20th day of pregnancy organization and maintenance of the nest, collecting through phone calls of 25 seconds every 2 minutes for and licking the offspring. Results: The indications of 12h. At birth, litters were weighed and standardized. clinical toxicity evaluated were not observed during Euthanasia proceeded on the 90th day of life. In serum the treatment period as the variations on body weight, were measured hormones related to metabolism as: feed and water intake, that were not significantly TSH, T3, T4, insulin, leptin and adiponectin. Statistics: different between the experimental groups. The Student’s t test (α=0.05). Results: No significant breastfeeding rats also did not demonstrated alterations differences were observed on hormones evaluated in its maternal behavior, presenting standard behavior either in male and female offspring. Conclusion: In the of lactating mother. Conclusion: The treatment with model studied the radiation does not alter the Valerian during lactation did not cause maternal hormones analyzed related to energetic metabolism. clinical toxicity and alterations on maternal behavior ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 99

Financial support / Acknowledgments: Universidade associated with tumor predisposition. The count of Federal de Juiz de Fora; CNPQ; Rede Mineira de cells, at optic microscopy, usually erythroblasts, is Bioterismo; Rede Mineira de Ensaios Toxicológicos e very time consuming. Therefore, automation is being Farmacológicos. searched and flow cytometry is one of the possibilities to optimize this analysis. In Brazil, there are no commercial available kits to detect micronucleus by EX 11 - GENDER DIFFERENCES IN flow cytometry and the antibodies and markers LOCOMOTOR ACTIVITY AND ANXIETY recommended are too expensive. Thus, the objective of PARAMETERS IN AN ADULT RAT MODEL OF this study was to validate the use of flow cytometry to MANGANESE SUBACUTE INTOXICATION score micronucleus in peripheral blood of mice using available reagents. Health young Swiss mice were PERINETTO, D.; EICHWALD, T. 1; SCHIMITZ, treated orally with cyclophosphamide 50 mg/kg bw C.R.R. 1; FLORES, M.V.B. 2; REMOR, A.P. 1; and demineralized water, at 24 hour intervals. The CARVALHO, D.1. blood was collected 42h after the last treatment of anesthetized animals. The erythrocytes were purified Universidade do Oeste de Santa Catarina, UNOESC- by density gradient with Percoll and the samples were Campus Joaçaba-SC, Brasil. fixed with paraformaldehyde. DNA of peripheral erythrocytes was stained using propidium iodide. tistical differences between groups in latency to fall in Reticulocytes were distinguished from accelerating Rotarod test. In EPM, ANOVA revealed a normochromatic erythrocytes by thiazole orange stain. significant reducing of percentage of time spent within The erythrocyte cell population was visualized on side open arms in male group that received 15mg/Mn/kg scatter area in logarithmic scale (SSC-A-log) versus compared to male control group (p<0.05). Differently, forward scatter area in linear scale (FSC-A-lin) there were a tendency (p=0.06) of increasing of (Guava, Merck). The gated cells (R1, R2 and R3) were percentage of time spent within the open arms for analyzed according to the intensity of fluorescence in 15mg/Mn/kg female group compared to female the FITC channel and in the propidium iodide channel. control. Differences between genders were found in A quadrant gate separated the four cell populations: 6mg/Mn/kg and 15mg/Mn/kg doses between males normochromatic erythrocytes, reticulocytes, and females for the same EPM parameter. There were micronucleated reticulocytes and micronucleated not statistical differences in other EPM parameters, normochromatic erythrocytes. Since the frequency of that include entries in closed arms (which evaluates micronuclei in negative control (demineralized water) locomotor activity) and time spent in central square was 0.01% and in positive control (cyclophosphamide) (which evaluates a decision-making in the EPM). was 0.03%, the comparison between controls Conclusion: The subacute model of Mn intoxication confirmed that the assay was successful. did not promote impairments in motor coordination, Financial support / Acknowledgments: Tecam balance and locomotor activity. However, it promotes Laboratórios - Tecnologia Ambiental e Merck S.A. dose-dependent alterations in anxiety levels of rats. Furthermore, this Mn intoxication model affects differently both genders, increasing anxiety levels in EX 13 - VALIDATION OF HEPATOTOXICITY males and decreasing these levels in females. INDUCTION MODEL BY CARBON TETRACHLORIDE: MALONDIALDEHYDE AND TRANSAMINASES AS BIOMARKERS EX 12 - FLOW CYTOMETRIC ANALYSIS OF IN VIVO MICRONUCLEUS ASSAY IN MELO, D. F. A. 1.; SILVA, M. A. C. 1; CUNHA, L. PERIPHERAL BLOOD: A NEW PROTOCOL C. C. 1, JERÔNIMO RAIMUNDO DE OLIVEIRA NETO.1. VIEIRA, C.O.1; ALVES, D.Z.1; SALVESTRO, T.Z.1; COSTA, M.C.2; ARAÚJO, I.F.1; PESTANA, C.B.1. 1 Universidade Federal de Goiás, Goiânia - GO, Brasil. 1 Tecam Laboratórios - Tecnologia Ambiental; 2 Merck S.A. Introduction: Carbon tetrachloride (CCl4) is capable of producing oxidative stress and cell lysis. This Micronuclei are small structures formed by intoxication mechanism has been used in pre-clinic chromosomes or fragments of chromosomes that have assays like liver damage induction model with high failed to be incorporated into the nucleus during the reproducibility and uniformity. The induction of liver cellular division. The mammalian in vivo damages becomes essential to investigate livers micronucleus test has been used to evaluate diseases, as well as to test new candidates for genotoxicity and high micronuclei levels have been hepatoprotective drugs. However, the period of ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 100 intoxication to developed cirrhosis may be so secondary metabolite with anti-inflammatory, extensive that it prolongs the suffering of experimental astringent, and antioxidant activities. Aimed: Identify animals. Objective: To develop a protocol of acute the potential of the dry standardized extract of A. intoxication by CCl4 that lasts less than 24 h, in vivo. tibourbou leaves (DSEAT) to protect the liver against Methods: 35 Wistar rats were distributed randomly in CCl4-induced hepatotoxicity in mice. Materials and 7 groups (5 animals each group). The group 1 Methods: The animals were treated with DSEAT (negative control) received olive oil, at a dose of 0.2 previously for 7 days, at doses of 25, 50, 100, 200, and mL / animal intraperitoneally (IP). The groups 2, 3 and 400 mg/kg and 18 mg/kg of rosmarinic acid; the liver 4 were intoxicated with carbon tetrachloride diluted in damage was induced by administering CCl4 olive oil (1:1 v/v), administered intraperitoneally, at intraperitoneally (i.p.) at days 3 and 7, 1 h before doses 0.5 mL / kg, 1.0 mL / kg e 2.0 mL / kg. The treating with DSEAT. The hepatoprotective activity groups 5 and 6 received 2.0 mL / kg of CCl4 diluted in was assessed using various biochemical assays such as olive oil (1:1 v/v), administered intraperitoneally, and alanine transaminase (ALT), aspartate transaminase the group 7 received no treatment. The groups 2, 3 and (AST), alkaline phosphatase (ALP), gamma- 4 were anesthetized and submitted to euthanasia after glutamyltransferase (GGT), malondialdehyde (MDA) 24 h and the groups 5 and 6 after 6 h and 12 h and histopatological studies. Approved by the Animal respectively. The blood was collected and 0,5 g of Research Ethics Committee of UFG no. 177/2011). liver excised. The hepatic intoxication was evaluated Results: DSEAT doses of 400, 200, and 100 mg/kg by measurement of malondialdehyde (MDA) levels in were not capable of protecting the liver against CCl4. plasma and liver homogenate by HPLC-PDA and the However, the dose of 50 mg/kg reduced AST by transaminases (ALT and AST) levels were measured 31.50% the dose of 25 mg/kg reduced GGT by 57.18% in plasma by spectrophotometry. The study was compared to the CCl4 (p < 0.05). In the liver, DSEAT approved by the Comissão de Ética para Uso de dose of 50 mg/kg and rosmarinic acid reduced MDA Animais/CEUA under No. 128/2014. Results: Among by 27.45% and 63.61%, respectively, whereas in the doses of CCl4 assessed after 24 hours of acute plasma, MDA was reduced in all the groups treated intoxication, the dose 2.0 mL / kg IP, increased the with DSEAT and rosmarinic acid. Conclusion: MDA and transaminases levels in plasma, but the DSEAT exhibits hepatoprotective effect only at low MDA did not increase in the liver. Otherwise, after doses and in vivo antioxidant activity after peroral 12h of acute intoxication, with the same dose of CCl4, administration. the levels of all the biomarkers increased, including Financial support / Acknowledgments: Conselho liver MDA. Conclusion: The CCl4 produces Nacional de Desenvolvimento Científico e intoxication, in vivo, in the period of 24h, being 12 h Tecnológico (CNPq), Fundação de Amparo à Pesquisa the best interval of time to quantify the biomarkers do Estado de Goiás (FAPEG), Fundação de Apoio à such as MDA in liver and plasma and to evaluate Pesquisa (FUNAPE) e Biotério Central da UFG. plasma activity of transaminases. Financial support / Acknowledgments: CNPq, FAPEG e UFG. EX 15 - A STUDY ON THE INFLUENCE OF MOCLOBEMIDE ON BASAL BLOOD PRESSURE AND ON TYRAMINE EX 14 - HEPATOPROTECTIVE AND ANTIOXIDANT ACTIVITIES OF DRY SARAIVA, E. R. 1; BORGES, D. 2; QUEIROZ, R. H. STANDARDIZED EXTRACT OF APEIBA C. 3. TIBOURBOU AUBL, IN MICE 1 Universidade de Rio Verde, Rio Verde - GO, Brasil; MELO, D. F. A. 1.; SILVA, M. A. C. 1; CUNHA, L. 2 Universidade de Rio Verde, Rio Verde - GO; 3 C. C. 1; MELO, D. S. 1; PARENTE, L. M. L. 1; Universidade de São Paulo, Ribeirão Preto - SP, CONCEIÇÃO, E. C. 2; LINO JÚNIOR, R. S. 3. Brasil.

1 Universidade Federal de Goiás, Goiânia - GO, Introduced on the market many years ago, the Brasil. moclobemide is a drug which has anti depressant properties due to selective and reversible inhibition of Introduction: Apeiba tibourbou, popularly known as type A monoamine oxidase ( iMAO–A ). This enzyme monkey comb, is found in Central America and in the is located in the mitochondrial membranes found in the Brazilian Cerrado region. The ethnopharmacological nervous terminals, the liver and other organs. Its uses of this species include treatment of rheumatism, activity is very important, regulating the metabolic spasms, and respiratory secretions. Moreover, the degradation of the catecholamines and, serotonin in the leaves are rich in antioxidant compounds, such as nervous tissue. Moclobemide is a benzamide-type flavonoids, tannins, and mainly rosmarinic acid, a iMAO-A which, regarding adverse reactions when ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 101 compared to clomipramine and imipramine, tricyclic on the cochlear function of Wistar rats. Material and antidepressants, causes fewer anti cholinergic Methods: 48 male Wistar rats, aged 60 days, were effects,such as orthostatic hypotension and vertigo. randomly assigned to 4 groups: Control (C - 12 rats The use of moclobemide, however, is still clinically exposed to water); Positive Control for auditory lesion restricted, mainly for the treatment of elderly patients, (PC - 12 rats treated with 24mg/kg intraperitoneal because of risks of hypertension crises when cisplatin); Dichlorvos (DV - 12 rats exposed to associated with dietary tyramine, even though it is a dichlorvos – 0.0015mg/mL); Cypermethrin (CY-12 type -A iMAO selective inhibitor. In order to rats exposed to cypermethrin – 1.667mg/mL). investigate possible blood pressure alterations caused Exposure to dichlorvos, cypermethrin and water by intravenous ministration of moclobemide, as well as occurred for 4 hours, 5 times/week, for 6 weeks. The increased tyramine´s pressor effects, the drug was amplitude of distortion product otoacoustic emissions given to normotensive rats, Wistar rats and before and after the exposure period was evaluated in spontaneously hypertensive rats, sorted into 10 groups 4,6,8,10 and 12 kHz. Body mass, clinical signs and ( n= 7 per group ). Also, a methodology of analysis plasma cholinesterase were also analyzed. Data were through liquid chromatography – which is highly presented as median and interquartile range or mean efficient for measuring the dosage of this drug in and standard error, and the groups compared by Mann Wistar rats and S.H.R. was adopted, so that the Whitney, Kruskal-Wallis or ANOVA. Results: There association between plasma concentration and the was no significant (p>0.05, ANOVA) change in effects of moclobemide were established. After relative body weight and plasma cholinesterase, injecting tyramine and tyramine associated with however, the animals showed transient clinical signs moclobemide, blood pressure was checked through a during exposure to DV (piloerection, dyspnea and cannula surgically placed in the carotid artery and the depression) and CY (piloerection, dyspnea, tremors alterations were recorded by a polygraph. The results and excitation). The frequencies of 8 and 10kHz were of this study show that moclobemide, in doses used in significantly affected bilaterally in the both pesticide the study, does not alone alter basal blood pressure, in groups (p < 0.05; Mann Whitney) and in CY group the Wistar rats and S.H.R., although when associated with frequency 12 kHz was affected bilaterally and also, the tyramine it does increase its pressor effect in a dose- frequencies 4 and 6 kHz on the right ear. These dependent manner and in S.H.R. the potentiation this alterations were more frequently manifested in effect was more intense. pesticide groups than in PC (12 kHz on right and 4 Financial support / Acknowledgments: FCFRP - kHz on left). Conclusions: Subchronic inhalation USP. exposure to dichlorvos and cypermethrin induced ototoxicity in rats without relevant systemic toxicity. Financial support / Acknowledgments: CAPES e EX 16 - OTOTOXICITY OF PESTICIDES IN FAPERGS WISTAR RATS

DALLEGRAVE, ELIANE; REIS, A.1; CUNHA, EX 17 - TOXIC EFFECTS OF SUNSCREEN E.O.1; DALMOLIN, S.P.2; BASSANI, D.S.2; OCTYL METHOXYCINNAMATE (OMC) MACEDO, M.B.1; SAUL, D.A.1; MACHADO, ADMINISTRATION DURING DEVELOPMENT M.S.1. OF RODENTS IN LACTATIONAL PERIOD

1 Universidade Federal de Ciências da Saúde de Porto GARCIA, E. B; BARBOSA, E. 1; FERRARIS, F. K. Alegre (UFCSPA) - Porto Alegre, Rio Grande do Sul, 1; SAVIGNON, T. 1; CHAVES, A. S. 1; Brasil; 2 Pontifícia Universidade Católica do Rio MUYLAERT, F. F. 1; DORO, L. H. 1; BRITO, T. M. Grande do Sul (PUC RS) - Porto Alegre, Rio Grande 1; MONSORES, N. F. 1; CARVALHO, V. F. 2, do Sul, Brasil. AMENDOEIRA F. C. 1.

Introduction: Pesticides are known for their impact on 1 Fundação Oswaldo Cruz, Instituto Nacional de health and the environment, representing a public Controle de Qualidade em Saúde, Rio de Janeiro - RJ, health problem. There are few studies researching Brasil; 2 Fundação Oswaldo Cruz, Instituto Oswaldo associations between hearing damage and pesticides, Cruz, Rio de Janeiro - RJ, Brasil. and Brazilian legislation does not regulate audiometric control of workers exposed to those substances, Introduction: The manganese (Mn) is an essential therefore it is necessary to execute a study in model metal, which plays important roles in different animal to verify the effects of pesticides in rat hearing. homeostastic processes. However, in cumulative Objective: To evaluate the effects of inhaled exposition it could lead to a Parkinson-like subchronic exposure to the organophosphorus neurological dysfunction, called as manganism. dichlorvos pesticide and the pyrethroid cypermethrin Objective: This work aimed to evaluate anxiety levels ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 102 and locomotor activity in adult males and females EX 18 - EVALUATION OF CRACK COCAINE Wistar rats that were submitted to a subacute model of SMOKE ON PREGNANT RATS: A METERNAL Mn intoxication. Methods: 19 male and 20 female EVALUATION AND TERATOGENIC adult Wistar rats were intraperitoneally (IP) injected 5 POTENTIAL times a week during 4 consecutive weeks with Mn chloride solution (MnCl2.4H2O) in 6mg/Mn/kg and SILVA, E.M.S. 1; ALVES R.B. 1; MENDES, P. F. 2; 15mg/Mn/kg doses. Control group received NaCl 0.9% SIMON, K. A. 1; HUEZA, I. M. 1,2. solution IP for the same period. After, the animals were tested in Elevated Plus Maze (EPM), in order to 1. Universidade Federal de São Paulo (UNIFESP), evaluate anxiety levels, and in accelerating Rotarod to Diadema - SP, Brasil; 2. Faculdade de Medicina evaluate locomotor activity and coordination. Results: Veterinária e Zootecnia da Universidade de São Paulo There were not found significant (FMVZ-USP), São Paulo - SP, Brasil. staINTRODUCTION: The UV radiation can lead to deleterious effects on human health therefore it’s Although there are several studies in animal models important to use sunscreen. OMC is an organic UV with the use of cocaine hydrochloride during filter used in cosmetics; and several studies have pregnancy, there are no reports in the literature about shown it as a potential endocrine disruptor of the the effect of crack smoke on fetal development. Thus, thyroid axis. The Central Nervous System is affected the present study aimed to evaluate maternal and fetal by the thyroid axis. Thyroid hormones have an parameters of the crack exposure before and during the important role in neuronal migration and maturation, entire gestation. It is known that it is common for the plus affect body structure development. OBJECTIVE: user to place cigarette ashes into the pipe to avoid the To evaluate whether OMC affects the thyroid axis and loss of the drug by melting it; thus, we used 30 female induces changes in neonatal development and immune rats that were divided into 3 equal groups: one exposed system in rodents during lactation period. METHODS: to 250mg of crack burned on a bed of cigarrete ashes Three groups of Wistar rats’ pups were exposed to (Ck group); one group exposed only to the burning of either corn oil, propylthiouracil (PTU) 4mg/kg/day or the ashes (Cc group) and a group that was only OMC 500mg/Kg/day, p.o. from PN5 to PN22. At this inserted in the burning apparatus just to mimicking the period, these animals were weighted, and development exposure procedure (Co group). All animals were parameters were observed. In PN23 some of the submitted to this experimental design 5-7days before animal’s blood were collected to T4 quantification; matching and during all the pregnancy twice a day. biochemical and hematological analysis. The Feed intake and body weight gain were performed remaining animals were tested for behavioral during all the experimental period. On the 21st day of alterations. To estimate OMC influence on the immune gestation, the females were anesthetized with xylazine system; lactating Swiss mice received either corn oil, and ketamine and submitted to cesarean section for PTU or OMC (250, 500 or 1000mg/kg/day), p.o. from exposure of the uterus and analysis of maternal PN1 to PN22. In PN23 we measured and quantified parameters and fetuses’ evaluation that were T4. Thymus and spleen were weighed and leukocytes performed according to classic teratogenicity were counted. RESULTS: OMC reduced weight gain protocols. Despite differences observed in maternal (p value < 0.001) and delayed eye opening of rats’ evaluation among the groups Co and Ck, as reduction pups (p value < 0.05) and of mice pups (OMC 1000 in the food intake (p<0.05, Co vs Ck), and diminished mg/kg/day; p value < 0.001 for weight and p value < non-gravid uterus (p<0.01, Co vs Ck), no statistical 0.05 for eye). In PN23, rats exposed to OMC had a changes were observed in the fetal parameters and reduction in total T4 levels (p=0.002). OMC also skeletal analysis among the groups evaluated. It is well reduced anxiety in female rats in EPM tests. Thymus known that crack consumption occurs compulsively, weight of the mice exposed to the OMC was reduced around 13 to 23 rocks by day, thus a possible lack of (p value < 0.001), but there was no interference in fetal results could be due to the low frequency of cellularity. However, the animals presented an increase exposure; however, it is important to highlight that in neutrophil counts (p <0.05). CONCLUSION: OMC classical protocols of teratogenicity: visceral and induces alterations in total T4 levels, reduces weight skeletal analysis could be not enough to better gain and delays eye opening. A milestone in characterize functional changes. Thus, it will be neurodevelopment, which may be correlated with the performed in our lab, a new experimental design to reduction of anxiety in EPM tests. However, OMC better evaluate postnatal changes promoted by high induces alterations in thymus weight and in levels of crack cocaine exposure than that here neutrophils. It suggests a deleterious influence on the employed. immune system. More studies are necessary to unravel Financial support / Acknowledgments: the OMC mechanism of action. FAPESP, grant number 2016/17050-9. Financial support / Acknowledgments: FIOCRUZ, INCQS, CAPES, PAPES/CNPq ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 103

EX 19 - EFFECTS OF REPEATED EXPOSURE possibly through its antioxidant action. Additionally, TO THE HERBICIDE ATRAZINE ON THE dopaminergic, serotoninergic and cholinergic systems NEUROBEHAVIORAL ACTIVITY OF RATS do not seem to be linked to the neurobehavioral AND ITS RELATION TO OXIDATIVE STRESS changes studied. AND NEUROTRANSMITTERS SYSTEMS Financial support / Acknowledgments: CAPES

ANSELMO, F.1; HORTA, D.F.1; BARRETTO, M.L.M.2; CALEFFI, J.T.1; BARBOSA, M.O.R.1; EX 20 - SUB-CHRONIC EMAMECTIN FERNANDES, A.A.H.3; GODINHO, A.F.1. BENZOATE EXPOSURE DISTURBS ANXIETY, MOTOR COORDINATION, AND BRAIN 1 São Paulo State University (UNESP), Institute of OXIDATIVE STRESS IN RATS Biosciences, Laboratory of Neurobehavioral Research, Botucatu, São Paulo, Brazil; 2 Federal Rural HORTA, D.F. 1; FRAIA, D. 1; ANSELMO, F. 1; University of Pernambuco (UFRPE), Recife, KAGA, A.K. 2; SILVA, C.S. 2; FERNANDES, Pernambuco, Brazil; 3 São Paulo State University A.A.H. 2; GODINHO, A.F. 1. (UNESP), Institute of Biosciences, Laboratory of Neurobehavioral Research, Botucatu, São Paulo, 1 São Paulo State University (UNESP), Institute of Brazil. Biosciences, Laboratory of Neurobehavioral Research, Botucatu, São Paulo, Brazil ; 2 São Paulo State Epidemiological studies have linked the pesticide University (UNESP), Institute of Biosciences, exposure with increase of several neurological Department of Chemistry and Biochemistry, Botucatu, disorders such as, anxiety, and also incidence of São Paulo, Brazil. neurodegenerative diseases like Parkinson's disease. Few and inconclusive studies address effects of the Emamectin benzoate, a pesticide of the avermectins atrazine (ATZ) on behavior. ATZ is used in many class, was initially introduced as a veterinary drug for cultures to increase productivity. The objective of this internal and external parasites control, and has being study was to evaluate the neurobehavioral activity of utilized in crops production. The action mechanism of anxiety, motor coordination and locomotor activity of avermectins class compounds is through interaction exposed animals, and the relationship with oxidative with GABA-gated chloride channels which is present stress, and dopaminergic, serotoninergic, and in vertebrates and mammals. Considering the potential cholinergic systems. Male adult Wistar rats received of avermectins class compounds for produce central during 28 days: no treatment (control-Ct), ATZ nervous system effects and the lack of information's 100mg/Kg (gavage), ATZ+Vitamin E (VE-200mg/kg, about EB toxicity to mammals present in open gavage), ATZ+levodopa (LEV-20mg/kg, gavage) and literature the aim of present work was evaluate effects ATZ+fluoxetine (FLU-10mg/kg, i.p.). At the end of of sub-chronic EB exposure on specific the treatments, the animals were tested in elevate plus- neurobehavioral parameters and brain oxidative stress maze (EPM-number of entries in the closed arms of in rats. Male Wistar rats received control (distilled the apparatus), in hole-board (HB-motor coordination water) or emamectin benzoate (0.5, 1, and 2 mg/kg) through of slip of paw inside of a hole), and in open treatments, via gavage, during 28 days. Behavior was field (OF-locomotor activity through of the number of assessed using open field, elevated plus maze, and square travelled). After the euthanasia, the activity of hole-board apparatus. Oxidative stress assay was brain cholinesterase was evaluated. In EPM, the performed assessing brain hydroperoxide lipid levels number of entries in the closed arms significantly and the antioxidant enzymes superoxide dismutase, increased in ATZ (7.68±0.49) in relation to the Ct catalase, and glutathione peroxidase. Overall, animals (5.08±0.46), which reflects increased anxiety. In HB, receiving emamectin benzoate presented anxyolitic- there was a significant increase in the number of slip of like behaviour and reduced motor coordination, paws in ATZ (4.20±0.39) in relation to Ct (2.08±0.35), accompanied by high hydroperoxide lipid levels and which reflects decreased motor coordination. The decreased antioxidant enzymes activity. The present group ATZ+VE had a significant decrease in the work demonstrated clearly that sub-chronic emamectin number of slip of paws (2.00±0.30) in relation to ATZ, benzoate exposure to rats alters anxiety and motor staying with average very close to the Ct. In OF, there coordination related to an increased oxidative stress were no significant alterations in locomotion. The status in brain tissue. These findings could strengthen assessment of brain cholinesterase activity, not shows the suggested relationship between pesticides exposure significant differences between the groups. The results and neurodegenerative diseases. Future directions for show that exposure to the ATZ increased anxiety, understand specific mechanism of action causing the impaired motor coordination and did not alter the effects here observed are necessary, including use of locomotor activity of animals. In the case of motor pharmacological strategies and research on coordination, VE reverted the injurious effects of ATZ, ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 104 neurotransmitters involved in the behavioral Financial support / Acknowledgments: Coordenação modulation. de Aperfeiçoamento Pessoal de Nível Superior Financial support / Acknowledgments: (CAPES) CAPES

EX 22 - RADIOPROTECTIVE EFFECT OF EX 21 - NEUROPROTECTIVE EFFECT OF THE SODIUM ALIZARINSULFONATE IN AN PHYTOCHEMICAL CARVACROL AGAINST EXPERIMENTAL MODEL OF ACUTE ACROLEIN-INDUCED TOXICITY RADIATION SYNDROME

SISTI, F. M. 1; SANTOS, N. A. 1; PETRI, C. 1; MACIEL, M.E.1,2, COSTANTINI, M.H.1,2, FERREIRA, R. 1; AMARAL, L. 1; SANTOS, A. 1. FORMOSA LEMOINE, F.1, LÓPEZ, G.D.3, MONTALTO, M.1, DÍAZ GÓMEZ, M.I.1, CASTRO, 1. Universidade de São Paulo, Ribeirão Preto - SP, J.A.1, CASTRO, G.D.1,2. Brasil. 1 UNIDEF, Centro de Investigaciones Toxicológicas Among recent findings about Alzheimer’s disease, (CEITOX), MINDEF-CONICET. CITEDEF. Juan B. acrolein has emerged as a new suspect. It is a highly de La Salle 4397. B1603ALO Villa Martelli, reactive aldehyde produced by cell metabolism and Argentina. E-mail: [email protected] 2 Instituto oxidative stress but also a ubiquitous environmental de Investigación e Ingeniería Ambiental, Universidad pollutant and drink/food component. An emerging Nacional de San Martín (UNSAM). Avenida 25 de consensus about this disease is the necessity to focus in Mayo y Francia. 1650 San Martín. Argentina. 3 early stages, as neuropathological changes were shown Laboratorio de Ensayos No Destructivos, División to occur many years before cognitive decline, memory Ensayos y Evaluación, Departamento Cabezas de loss and diseases diagnosis. In the earliest pathological Combate. CITEDEF. Juan B. de La Salle 4397. changes, neurite dystrophy and loss occurs as B1603ALO Villa Martelli, Argentina. cytoskeletal abnormalities, resulting in synaptic failure. Acrolein toxic effect as neurite outgrowth Increased intracellular calcium is a factor known to be impairment is still unknown. Regeneration and involved in the process leading to cell death. Changes outgrowth of neurites may recover brain in calcium homeostasis are relevant to the late stages communication and cognitive functions, therefore, of cell injury. Sodium alizarinsulfonate (ASR) has a compounds able to induce neuronal projections might potent action as calcium chelator, thus interfering with have beneficial effects in neurodegeneration. Carvacrol the entrance and action of this element in the cell. It is is a phytochemical widely used as food additive whose our interest to develop less toxic radioprotectors, either neuroprotective potential was never explored. Thereby, by themselves or as adjuvants to drugs already the aim of this study was to evaluate the effect and approved for their use in humans but having toxic side mechanisms of acrolein and carvacrol on effects. In this work, using an experimental model of neuritogenesis process. For this, PC-12 cells were Sprague-Dawley rats (both sexes) exposed to X treated with acrolein, carvacrol, nerve growth factor radiation (2 Gy, whole body), the radioprotective (positive control), tropomyosin receptor kinase A effect of ASR was evaluated. Groups of 8 rats were (trkA) inhibitor (K252a) and phosphatidylinositol 3- exposed at a dose of 2 Gy. At 48 hours post exposure kinase (PI3K) inhibitor (LY294002). Neurite blood samples were obtained by tail puncturing, outgrowth was assessed by inverted-contrast-phase followed by sampling at 7, 14, 21, 28 and 60 days. The microscopy and the Image J open source software. Our erythrocyte, leukocyte and leukocyte formula were results show that acrolein affects neurite outgrowth counted. Survival curves up to 60 days were also reducing the percentage of differentiated cells. On the performed. Genotoxic effects in leukocytes were other hand, carvacrol stimulated neuritogenesis, effect assessed by the Comet assay (one hour post suppressed by the inhibitors. The neurite outgrowth irradiation). The effects of ASR were tested following induced by carvacrol, involving the activation of its administration as a single dose of 100 mg/kg (i.p.), trkA/PI3-K pathway, suggests that it is capable of one hour before irradiation. In the irradiated animals mimic neurotrophic factors, thus controlling erythrocytes were depleted (females, p<0.01), and the neurogenesis, neuronal plasticity and survival. white blood cell count was drastically reduced with Furthermore, carvacrol was able to prevent the respect to the control (both sexes, p<0.01), also reduction of differentiated cell’s percentage caused by presenting an altered formula. Genetic damage acrolein. In conclusion, our results suggest that revealed by the Comet assay was significantly reduced acrolein can be involved in the earliest alterations of by treatment with ASR (p<0.01). The effect of ASR on Alzheimer’s disease and that carvacrol may be a new blood parameters tested was protective in the recovery therapeutic agent to be explored. of erythrocytes in females (p<0.01). No statistically ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 105 significant protection was observed in the recovery of uterus from rats treated with standard Lieber & De the leukocyte level or the leukocyte formula (both Carli liquid diet for 28 days revealed extensive sexes). However, a significant protective effect was vacuolization in cytoplasm and loss of cell content. In observed for survival (p<0.05). These findings suggest addition, we observed the promotion of oxidative that ASR, either by its known action as calcium stress as evidenced by increased response in the t- chelator or by scavenging free radicals sparked by butylhydroperoxide induced chemiluminiscence and ionizing radiation, has a good potential as the depletion of the protein sulfhydryl content. Minor radioprotective agent by itself or as adjuvant to but significant depletion were observed in the GSH amifostine (WR-2721). This drug is an effective content, accompanied by a tendency to decrease in the radioprotector but has important toxic effects per se GSH/GSSG ratio. A highly significant finding was the that prevent its repeated use. diminished activity content of glutathione peroxidase. Financial support / Acknowledgments: PIDDEF Results suggest that in the rat uterus, metabolism of 11/12 (MINDEF, Argentina). ethanol to acetaldehyde and oxidative stress may play a role in alcohol effects on female reproductive function. EX 23 - OXIDATION OF ETHANOL TO Financial support / Acknowledgments: PIP ACETALDEHYDE AND OXIDATIVE STRESS 11220150100067CO (CONICET, Argentina). COULD BE INVOLVED IN THE ALCOHOL DRINKING PROMOTED CELL INJURY OF THE RAT UTERUS EX 24 - EVALUATION OF PSITTACANTHUS CORDATUS (HOFFMANNS.) G. DON MACIEL, M.E.1,2, COSTANTINI, M.H.1,2, TOXICITY FORMOSA LEMOINE, F.1, CASTRO, G.D.1,2. SILVA, H. V. B. C. 1; CÔRTES, A. L. A. 1; VIANA, 1 UNIDEF, Centro de Investigaciones Toxicológicas G. F. S. 1. (CEITOX), MINDEF-CONICET. CITEDEF. Juan B. de La Salle 4397. B1603ALO Villa Martelli, 1 Faculdade Guanambi, Guanambi - BA, Brasil. Argentina. E-mail: [email protected] 2 Instituto de Investigación e Ingeniería Ambiental, Universidad Introduction: Psittacanthus cordatus (Hoffmanns.) G. Nacional de San Martín (UNSAM). Avenida 25 de Don is a parasitic plant used empirically to treat Mayo y Francia. 1650 San Martín. Argentina. pharyngitis, but lacking scientific evidence to prove its safety. Objective: The present study aims to evaluate It is known that alcohol drinking can lead to the toxicity of the hydroalcoholic extract of P. impairment in reproductive function in women. cordatus. Materials and Methods: Dehydrated and Controversial studies from others suggested that pulverized P. cordatus samples were submitted to a alcohol intake could be associated with some maceration procedure for the production of the deleterious effects in the uterus. Not all the effects of hydroalcoholic extract. The extract was rotoevaporated alcohol drinking on female reproductive organs can be and oven dried to remove all solvent. Toxicity explained in terms of endocrine disturbances. assessment was performed by the Artemia salina assay Deleterious effect of alcohol or its metabolites in situ using P. cordatus sample from a Bahia semi-arid city. could also play a role. In this study we analyze the Briefly, ten nauplii of A. salina were placed in contact possibility that part of these effects were mediated with different concentrations (3000, 750, 375 and through alterations of uterus function related to ethanol 187.5 mg/L) of the solubilized extract in synthetic oxidation to acetaldehyde occurring in that tissue. We seawater. Potassium dichromate and synthetic seawater found a metabolism of alcohol to acetaldehyde in the were used as positive and negative controls, rat uterus tissue cytosolic fraction mediated by respectively. The absence of nauplii mobility was xanthine oxidoreductase, requiring a purine cosubstrate evaluated in 24h and 48h. All procedures were and inhibited by allopurinol. This activity was detected performed in triplicate. The number of dead nauplii by histochemistry in the epithelium and aldehyde was quantified and expressed in mean among dehydrogenase activity was detected in the muscular triplicates. Descriptive statistical analyzes were layer and in the serosa. There was also a microsomal performed and the mean lethal concentrations (LC50) process, not requiring NADPH and of enzymatic were calculated according to the exposure time. nature, oxygen-dependent and inhibited by Results: The results showed an increasing linear diethyldithiocarbamate and diphenyleneiodonium. relationship between extract concentration and mean After alcohol exposure through a standard Lieber and number of dead nauplii for 24h (y=0.0019 + 1.9211; De Carli diet for 28 days, a severe atrophy in the rat R2=0.9567) and 48h (y=0.0022 + 2.7098; R2=0.9620) uterus was observed, accompanied by significant of exposure. The CL50 for 24h and 48h were, alterations in its epithelial cells. Ultrastructure of respectively, 1620.5 and 1041.0 mg/L, showing ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 106 absence of toxicity by the parameters validated for the of the city, the LC50 in 24h and 48h were, test. Conclusion: These results suggest absence of respectively, 5407 mg/L and 1077.6 mg/L, toxic potential of the hydroalcoholic extract of P. demonstrating absence of toxicity. Conclusion:These cordatus when evaluated by A. salina assay. However, results suggest absence of toxic potential of the since the A. salina assay is considered a preliminary hydroalcoholic extract of P. oleracea when evaluated toxicity study, further analysis is needed to corroborate by A.salina assay. these findings. Financial support / Acknowledgments: Faculdade Financial support / Acknowledgments: Faculdade Guanambi Guanambi

EX 26 - EFFECTS OF AIR POLLUTION OVER EX 25 - EVALUATION OF THE CYTOTOXIC LIVER, KIDNEY AND LUNG OF RATS POTENTIAL OF THE HYDROALCOHOLIC EXTRACT OF PORTULACA OLERACEA RIBEIRO, J.P.; KALB, A.C. 1; MAYA, S.B. 1; TESTED BY ARTEMIA SALINA ASSAY GIODA, A. 2; MARTINEZ, P.E. 1; MONSERRAT, J.M. 1; GIODA, C.R. 1. SILVA, J. B. D.; FERREIRA, A.S.1; JUNIOR, R.C.M. 1; VIANA, G.F.S.1. 1. Universidade Federal do Rio Grande, FURG, Rio Grande, RS, Brazil; 2. Pontifícia Universidade 1. Faculdade Ganambi, Guanambi - BA, Brasil. Católica do Rio de Janeiro (PUC-Rio), Rio de Janeiro, RJ, Brazil. Introduction: Portulaca oleracea is a plant that develops easily in different soils, climates and places. Particulate matter (PM) presents in its constitution It is used in several regions with food and therapeutic different chemical substances that have been purpose, although its safety has not yet been evaluated. associated to health problems and increased risk of Objective: This research aimed evaluate the cytotoxic mortalities due to its toxicity. In this study, fine potential of the hydroalcoholic extract of Portulaca particulate matter (PM2.5) samples were collected in a oleracea. Materials and Methods: Dehydrated and rural region (RR) and in an industrial region (IR) and pulverized P. oleracea samples were submitted to a rats were exposed to the PM2.5 extracts at different decoction procedure for the production of the concentrations (concentrated (CS), 5 times diluted (5x) hydroalcoholic extract. The extract was rotoevaporated and 10 times diluted (10x)) for 25 days. The objective and oven dried to remove all solvent.The evaluation of of this experiment was to verify its potential toxicity in the cytotoxic potential was performed by Artemia liver, kidney (cortex and medulla) and lung and salina assay with samples of P. oleracea from three whether there was difference in parameters analyzed cities in the semi-arid region of Bahia (Brumado, between regions with different pollution indexes. Caetité and Guanambi). Briefly, tennauplii of A.salina Biochemical analysis as total antioxidant capacity were placed in contact with different concentrations of (ACAP), lipid peroxidation (LPO), activity of the solubilized extract in synthetic seawater. Potassium glutamate cysteine ligase (GCL), reduced glutathione dichromate and synthetic seawater were used as (GSH) levels and activity of glutathione S-transferase positive and negative controls, respectively. The (GST) were investigated and the experiment was absence of nauplii mobility was evaluated in 24h and approved by the Ethics Committee of the University 48h. The number of dead naupliiwas quantified and (CEUA-FURG) (protocol n. P075/2014). Overall, the expressed in mean among triplicates. The mean lethal results showed that liver, kidney and lung have concentrations (LC50) were calculated according to undergone changes in ACAP, increased of LPO and the city of origin of the plant. The comparison of the decreased in GSH levels in different concentrations of mean number of dead nauplii among groups was PM2.5 extracts. In the liver was observed a significant performed by ANOVA and Tukey post hoc test, being increase in GCL (IR_5x, and CS groups) and GST accepted as significance the value of p<0.05. Results:It activities in RR_5x and CS in both regions when was observed, especially after 24 hours of exposure compared to controls. This study evidences, for the and regardless of the place of origin of the plant, low first time, the different responses of particulate matter mortality of nauplii even at the highest concentrations in renal cortex and medulla. In renal cortex, GCL of the extract. For the highest concentration evaluated activity was decreased in most of the groups tested and 48h of exposure, the number of dead nauplii was while GST activity was increased only in 5x groups in significantly lower for Caetité when compared to both regions. On the other hand, in renal medulla, GCL Brumado and Guanambi. The LC50 in 24h and 48h activity decreased for RR_10x and IR_ CS but were, respectively, 5111 mg/L and606.5 mg/L for increased in RR_5x and GST activity was increased in Brumado, 2833 mg/L and 2833mg/L for Caetité and RR_10x, IR_5x and IR_CS groups. Lung glutamate 444 mg/L and1143.5 mg/L for Guanambi. Regardless cysteine ligase (GCL) increased for concentrated ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 107 extracts, in both regions. Moreover, this tissue also texture of the organs analyzed. Conclusion: This study showed an increase in the GST activity when higher indicates that the dry leaf extract does not cause acute metal concentrations were present (5x and CS). Our oral toxicity effects at the doses tested. According to data suggest that soluble portion PM2.5 could change the classification of acute toxicity recommended by redox status and to leave tissues injuries of organ OECD, this extract can be categorized into class 5 considered target and not target in rats from areas (LD50> 5.000 mg/kg). different pollution index. Financial support / Acknowledgments: PBIC/UEG Financial support / Acknowledgments: CNPq, and PIBIC /CNPq (Process # 154075/2015-9). CAPES, and FAPERJ.

EX 28 - ACUTE ORAL TOXICITY OF THE DRY EX 27 - ACUTE ORAL TOXICITY OF THE DRY EXTRACT OF JUSTICIA PECTORALIS JACQ. LEAF EXTRACT OF MOMORDICA (ACANTHACEAE) CHARANTIA LINN (CUCURBITACEAE) IN WISTAR RATS. LIMA, A.R.S. 1,4; COSTA, T.L.C. 2,4; SOARES, J. A. 2,3; TRAUTENMULLER, A.L. 2,3; LÔBO, L.A.C. SOARES. J. A. 1; TRAUTENMULLER, L.A. 1; 2,4; AMARAL, V.C.S. 1,3; PAULA, J.A.M 1,4. GODOI, M.M. 1; PAULA, J.A.M. 1; AMARAL, V.C.S. 1. 1. Post-Graduate Program in Applied Sciences to Health Products, Campus of Exact and Technological 1 Universidade Estadual de Goiás, Anápolis - GO, Sciences, State University of Goiás (UEG), Anápolis - Brasil. GO, Brazil; 2 Pharmacy Course, Campus of Exact and Technological Sciences, UEG, Anápolis - GO, Brazil. Introduction: Momordica charantia L., popularly 3 Laboratory of Pharmacology and Toxicology of known as bitter melon or “melão-de-são-caetano”, is a Natural and Synthetic Products, Campus of Exact and perennial climber belonging to the family Technological Sciences, UEG, Anápolis - GO, Brazil; Cucurbitaceae. This plant is widely used due to its 4 Laboratory of Botanical, Chemical and Biological biological properties, including anti-diabetic, anti- Studies of Medicinal Plants, Campus of Exact and cancer, anti-inflammatory, anti-bacterial, antifungal, Technological Sciences, Anápolis - GO, UEG Brazil. anti-viral, anti-helminthic, anti-oxidant, and anti- hyperlipidemic activities. The fruit, leaves, seeds and Introduction: J. pectoralis is an herb of medicinal roots of this species has long been consumed as food interest employed in folk medicine in Latin American and medicine in many countries, including Brazil. The countries as an analgesic, anti-inflammatory and therapeutic potential of this plant has been studied sedative. It is known for properties of its coumarins extensively, focusing on its fruit and seeds. In this 1,2-benzopyrone and umbelliferone. This species is on context, studies have demonstrated relatively low the list of plants of interest to the Brazilian Health toxicity of the M. charantia L. fruit when orally used. System. Although there is broad utilization of this However, studies have reported that seeds are toxic medicinal plant, there is insufficient data about its and can induce infertility in rats. Despite the extensive potential toxicity. The toxicity data are considered literature on the pharmaceutical potential of this mandatory by the regulatory agencies. Objective: The medicinal plant, there are few studies regarding the aim of the study was to evaluate the acute oral toxicity toxicity of leaves of this species. Objective: This study of the dry extract (DE) of aerial parts of J. pectoralis in aimed to evaluate the acute oral toxicity of the dry leaf Wistar rats. Materials and Methods: The aerial parts of extract of M. charantia L. in rats. Material and J. pectoralis were collected in the Garden of Medicinal Methods: The experimental design was developed Plants of the Goiânia Botanic Garden, dried (40 °C) based on the Organization for Economic Cooperation and pulverized. The liquid extract was prepared by and Development (OECD), protocol 423. Female percolation in ethanol 20% (w/w) and spray dried. The Wistar rats (n=6/groups) were orally treated with the acute oral toxicity testing was carried out under the vehicle or the dry leaf extract of M. charantia L. at Organization for Economic Cooperation and dose levels of 300 or 2.000 mg/kg. The animals were Development, guideline no 423. Female Wistar rats monitored daily for mortality, behavior and appearance (n=6 per group) were orally treated with the vehicle or within the first 8 hours and for 14 days after the the dry aerial parts extract of J. pectoralis (DE), administration of the extract. At the end of the study, standardized on coumarins, at dose levels of 300 and the animals were necropsied for analysis of spleen, 2000 mg/kg. Behavioral responses were observed liver and kidneys. Results: There were no clinical signs within the first 8 hours and for 14 days after the of toxicity and no deaths in any group throughout the administration of DE. Body weight gain, food and experimental period. Moreover, macroscopic water consumption were recorded daily for 14 days. examination revealed no abnormalities in the color or On the 14th day the animals were necropsied for ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 108 analysis of liver and kidneys and the weights were development and fertility were evaluated. The testes expressed as relative mass. Results: During the were made by ANOVA, with p<0.05 as a level of experimental period there was no death of any animals. significance. Results: Silymarin treatment did not The clinical signs were absent for all doses. The affect the frequency of estrous cycle among the kidneys and liver are responsible for the metabolism groups: diestrus (p = 0.162); estrus: (p = 0.100); and and excretion of xenobiotics, but the autopsy did not proestrus: (p = 0.671). Weight gain during the non- show significant macroscopic and weight changes of pregnancy phase: (p <0.001); in pregnancy: (p = the organs of the test group compared to the control 0.012) and lactation: (p = 1,000) were did not different group (p>0.05), suggesting low toxicity of the test between the groups. Fertility analysis did not show species. Conclusion: This study shows the low acute difference among the groups, total pups (p = 0.495); oral toxicity of DE J. pectoralis, with LD50 estimated number of females (p = 0.789); number of males (p = between 2000 and 5000 mg/kg. 0.457). Conclusion: Silymarin treatment did not Financial support / Acknowledgments: To the interfere in the estrous cycle and did not produce toxic Garden of Medicinal Plants of the Goiânia Botanic effects for the gestational development at the doses Garden for plant supplies, Fundação de Amparo a proposed by this study. Pesquisa do Estado de Goiás (FAPEG) and Financial support / Acknowledgments: CNPq, Universidade Estadual de Goiás (UEG) for financial Capes, Fapeg support.

EX 30 - SOLANUM DIPLOCONOS EX 29 - EVALUATION OF GESTATIONAL (SOLANACEAE) FRUITS: IN VITRO AND IN SECURITY OF SILYMARIN IN THE PRE- VIVO TOXICOLOGICAL EVALUATION GESTATIONAL PHASE UNTIL LACTATION VENTURI, I. 1; PAVAN, S.E. 1; FRATONI, E. 1; SANTOS, J.A. 1; BARBOSA, C.C. 1; SOUZA, M.R. THIESEN, L.C. 1; SOUZA, M.M. 1; MALHEIROS, 1; NUNES, N.J.N. 1; NISHIMURA, A. 1; ARAUJO, A. 1; SANTIN, J.R. 1. A.A. 1; MORAIS, L.C.A. 1; COSTA, S.J.1; COSTA, R.M. 1. 1. Programa de Pós-graduação em Ciências Farmacêuticas - Universidade do Vale do Itajaí, SC, 1 Universidade Federal de Goiás, Goiânia – GO. Brazil.

Introduction: Silymarin or Silybum marianum is an Introduction: Solanum diploconos (Solanaceae) fruits herbal medicine used in treatment of liver diseases, are consumes, but there are no reports about their property already proven in clinical studies. In the toxicity in the literature. Objective: In this way, the Silymarin extract are present at least seven flavonoids present study demonstrates for the first time the in vivo with antioxidant, anti-inflammatory, antifibrotic, and in vitro toxicological effects of fruits. Materials detoxifying and regenerative activities. Non-alcoholic and Methods: In vitro toxicological evaluation was fatty liver disease, characterized by fatty infiltration in performed by hemolytic activity assay, cell viability the liver with high evolutionary potential, is common and micronucleus test. In vivo acute (2000 mg/kg) and today. This disease has a prevalence in both sexes, and sub-acute (30, 100, 300 mg/kg) toxicological tests (28 has as one of the risk factors the polycystic ovary days) were performed. In these tests hematological and syndrome. Thus, pregnant women may be susceptible biochemical parameters were also evaluated besides of liver disease, there is an estimative that 3% are the histopathological analysis of the kidney and liver. affect by these pathologies, and when severe, are The locomotor activity was evaluated through the associated with the mortality and morbidity of the Open field and Rota rod tests. Results: The results mother and fetus. The liver disease in pregnancy may found in the acute and sub-acute toxicology trials be a new reaction of maternal organism or related to a demonstrate that the fruits extract did not promote pre-existing disease, and even with the diagnosis and behavior alterations, did not alter the water and feed therapy advancements, there are still difficulties in consumption and animal weight. No changes were discover and treating. Objective: evaluate the effects of observed in the hematological and biochemical Silymarin exposure during pre-gestational, gestational parameters. The extract did not promote changes in and lactation periods. For these, sixty female Swiss motor activity and liver or kidney histopathological mice were used, distributed in five groups: Testimony, changes, not show hemolysis of the erythrocytes and Control, Sil1, Sil2 and Sil3 (all with n=12). not induce mutagenicity or cellular cytotoxicity. Methodology: Control animals received, o.p., Saline. Conclusions: Together the findings provide valuable (5 ml/kg) and the animals treated with Silymarin data on the acute, sub-acute and mutagenic oral received the extract, o.p., (100 mg/kg, 200 mg/kg, 300 toxicological profile of extract obtained from S. mg/kg). Estrous cycle, body mass gain, gestational ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 109 diploconos fruits, which shown to be safe for acute and does not promote acute toxicological alterations. sub-acute administration. Conclusions: Data obtained point out that the Financial support / Acknowledgments: The authors hydroethanolic extract of L. chinensis leaves presents thank the Conselho Nacional de Desenvolvimento hepatoprotective activity, corroborating with the Científico e Tecnológico (CNPq), Fundação de Apoio ethnopharmacological use, and does not present acute à Pesquisa Científica e Tecnológica do Estado de Santa toxicological effects. Catarina (FAPESC) and the University of Vale do Itajaí (UNIVALI) for their financial support. Financial support / Acknowledgments: The authors are thankful to Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, PVE, grant EX 31 - ACUTE TOXICITY AND 88887.122964/2016-00), Conselho Nacional de HEPATOPROTECTIVE EVALUATION OF Pesquisa e Tecnologia (CNPq, grant n. 444682/2014-7 LITCHI CHINENSIS LEAVES EXTRACT and 483577/2013-8), and FAPESC (Fundação de Amparo à Pesquisa e Inovação do Estado de Santa SANTIN, J. R.; THIESEN, L.C. 1; NUNES, M.L.O. 2; Catarina) (Edital PRONEM, grant 2708/ 2012) by MEYRE-SILVA, C. 2; PASTOR, V.D. 3; FALONI, financial support and CAPES for providing financial S.F. 1; COUTO, A.G. 1; SILVA, L.M. 1; BRESOLIN, support in the form of a doctoral's degree scholarship T.B. 1. to Thiesen, L.C.

1 Postgraduation Program in Pharmaceutical Sciences, Itajaí, SC, Brazil; 2 Núcleo de Investigações Químico- EX 32 - TESTOSTERONE INDUCES Farmacêuticas (NIQFAR), Itajaí, SC, Brazil; 3 UTEROTROPHIC EFFECT IN IMMATURE Pharmacy Course, Universidade do Vale de Itajaí RATS BY AROMATASE CONVERSION INTO (UNIVALI), Itajaí, SC, Brazil. ESTROGEN

Introduction: L. chinensis is popularly known as MÜLLER, J. C. 1; SARGAÇO, R. M. 2; GRECHI, N. cherry, lichee, litchi, lychee, mountain lychee, water 2; BARBOZA, L. N. 3; DALSENTER, P. R. 2; lychee and in Brazil as Lichia. The infusion of its BOARETO, A. C. 2. leaves is used in the Chinese folk medicine as a liver tonic. Objective: This study was designed to evaluate 1. Faculdades Pequeno Príncipe, Curitiba - PR, Brasil; the hepatoprotective and acute toxicological effects of 2. Universidade Federal do Paraná, Curitiba - PR, hydroethanolic L. chinensis leaves extract in HepG2 Brasil; 3. Universidade Federal da Grande Dourados, cells and in mice. Material and methods: The Dourados - MS, Brasil. cytotoxicity and hepatoprotective activity of L. chinensis leaves extract were evaluated in HepG2 cells Introduction: Considering the key role of aromatase in and MTT assay, hepatic injury markers aspartate reproductive processes and its involvement in the aminotransferase (AST), alanine aminotransaminase development of diseases with high levels of incidence (ALT), lactate dehydrogenase (LDH) leakage were and metastatic potential, it is clear the importance of performed. In vivo, hepatoprotective activity of L. developing screening tests for detect substances able of chinensis against carbon tetrachloride (CCl4)-induced altering this enzyme. The human (recombinant) acute liver injury was evaluated in mice. The acute microsomal aromatase assay detects substances that toxicity was also evaluated in mice. Results: The L. can inhibit aromatase activity, however, it does not chinensis extract was able to restore the viability in detect substances that induce aromatase activity and cells treated with CCl4 in all concentrations evaluated. substances that increase or suppress the protein levels In addition, the extract reestablished the AST levels. of this enzyme. Besides that, there is no specific in The administration of CCl4 in mice promotes, cell vivo test to evaluate changes in the aromatase enzyme, swelling in the liver tissue, loss of sinusoid capillary which would also involve the pharmacokinetics and spaces and structural disarrangement of the hepatic dynamics of the endocrine system. Matherial and lobe, apoptosis and leukocyte infiltration. The CCl4 Methods: For standardization of an animal model for promotes an increment in the activity of superoxide the evaluation of interferences on aromatase activity, is dismutase (SOD), catalase (CAT) and glutathione require a model with no hormonal interference by the redutase. In addition, the CCl4 evokes an increase in hypothalamic-pituitary-ovarian axis cyclicity, but at the alanine aminotransferase (ALT) in the serum, the same time already expresses aromatase in target Tumor Necrosis Factor (TNF) and Interleukin-6 (IL-6) tissues. Thus, the animal model consisted of immature levels in the hepatic tissue. The extract does not female rats, which are not yet cycling, but already modify SOD, CAT, GSH and GST profile. However, it expressed the aromatase enzyme in the large antral reduces the inflammatory parameters TNF and IL-6, follicles. Thenceforth, the first step in the and reestablishes the morphology of hepatic tissue, and standardization of this model was to evaluate whether ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 110 administration of testosterone plus flutamide could be renovation was 100%/day by dripping regulated with converted into estradiol by aromatase, in sufficient equipment for physiological serum. In addition, an concentration to produce uterotrophic effect, in a test aerator bubbled air constantly maintained the like the uterotrophic assay. In addition, uterine oxygenation of the water/solution in the experimental histomorphometry was performed to evaluate whether aquariums. The cholinesterase activity dosage was the trophic effect would be related to morphological made using a specific kit (Doles). The sample changes estradiol like those produced by estradiol. consisted in the supernatant of the body of each fish Results and Conclusion: The uterotrophic and without the liver and gill, homogenized with saline morphological effects observed by treatment with 0.9%. RESULTS: Cholinesterase activity in fish testosterone plus flutamide indicates that part of treated with methamidophos, glyphosate and testosterone was converted into estradiol by aromatase, mancozeb did not change. The values of enzyme and the estradiol produced such effects by acting on activity (UI/mL) for the different groups were: Control uterine estrogen receptors. These preliminary results 7.08 ± 0.63; Metamidophos 10.99 ± 0.55; Glyphosate showed that there is sufficient aromatase on the ovaries 10.66 ± 1.93 and Mancozeb of 10.99 ± 3.46. of immature rats to form a basis on which to develop CONCLUSION: The maximum value determined as an in vivo test to identify aromatase inhibitors based safe by Brazil Health Ministry for pesticide water on the uterotrophic assay, and relate it to in vivo and in contamination does not promote changes in the vitro tests well standardized. cholinesterase activity of fishes exposed during 30 Financial support / Acknowledgments: Fundação days to methamidophos, glyphosate and mancozeb. Araucária; CAPES Financial support / Acknowledgments: Universidade Estadual de Goiás Edital Interno CCB 001/2016 - PVIC-UEG. EX 33 - EFFECTS OF THE INTOXICATION WITH XENOBIOTICS IN THE ACTIVITY OF CHOLINESTERASE IN POECILIA EX 34 - REFLEXOLOGICAL AND PHYSICAL RETICULATA DEVELOPMENT OF WISTAR RAT OFFSPRING EXPOSED TO PSYCHOTRIA COLORATA BORGES, K. R. S. B.; REZENDE, L. P.; ALKALOIDIC EXTRACT DURING VERDIONO, W. L.; SILVA, N. O. D.; SANTOS, D. PREGNANCY AND LACTATION R.; MONTEIRO, J. A.; VIEIRA, D. R.; DIAS JUNIOR, W. JAVAÉ, N. R. K. 1; MOURA, L. T. S. 1; VASCONCELOS, I. C. 1; MARIANO-SOUZA, D.P. Universidade Estadual de Goiás 2; PAIVA, J. P. 3; MARUO, V. M. 1.

INTRODUCTION: The aquatic environment is often 1. School of Veterinary Medicine and Animal affected for contamination of substances that can bring Sciences, Federal University of Tocantins, Araguaína - problems to the aquatic animals and consequently to TO, Brazil; 2. Faculty of Biology, Federal University the humans beings. The xenobiotics are essential for of Tocantins, Araguaína - TO, Brazil; 3. Faculty of the agriculture, but the inappropriate manipulation can Chemistry, Federal University of Tocantins, Araguaína take it to streams, springs and rivers. The washing of - TO, Brazil. equipment used in the pulverization and derivation for the wind during the application by air are the main Introduction: Psychotria colorata, plant of the means of transport to the water, beyond leaching. Rubiaceae family, is commonly known as Perpétua do OBJECTIVE: To evaluate the effect of xenobiotics in mato, and used as analgesic among Amazonian cholinesterase activity of Poecilia reticulata treated for caboclos. Pharmacological analysis of P. colorata 30 consecutive days. MATERIALS AND METHODS: leaves extracts revealed opioid-like analgesic activity The fishes used in this work were from the aquatic attributed to pyrrolidinoindoline alkaloids. Objective: animal husbandry of Physiological and Biochemical Investigate the effects of administration of P. colorata Toxicology Laboratory of Goiás State University, leaves alkaloidic extract (EAL) on reflexological and Campus Ceres. The animals were separated into 4 physical development of Wistar rat offspring. Material groups: Control, Methamidophos (12μg/L), and methods: The leaves of the plant were dried at Glyphosate (500μg/L) and Mancozeb (180μg/L). The 55°C, moistened with NH4OH 6N, extracted with doses were obtained from the Allowed Maximum ethanol in a Soxhlet apparatus and concentrated in Value described in Portaria n.2914 (12/12/2011) from rotary evaporator at 45°C to obtain EAL. Female rats Brazil Health Ministry. The density of animals in were divided in two groups (n = 20), an experimental aquaria was 3fishes/2liters, totaling 24 fishes, group treated with EAL 238 mg.kg-1 body weight distributed equally in males and females, diluted in 5% Tween 80, and control group that 6fishes/intoxication group. The water/solution aquaria received only the vehicle. Treatments were ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 111 administrated by gavage from 6th day of gestation Introduction: Psychotria colorata (Rubiaceae) is a plant (GD6) up to the post natal day 21 (PND 21), in used as analgesic in popular medicine. Cattle ranchers agreement with the protocols of EPA 19961 and from the state of Tocantins reported that the plant is OECD 4262. The postnatal evaluation of rat offspring abortifacient. Objective: Investigate the effects of the was assessed in one male and female pups from each administration of the P. colorata ethyl acetate extract litter observed for the following parameters of physical on the reproduction of female Wistar rats. Material and development: hair growth, pinna detachment, eruption methods: The leaves of P. colorata were collected in of incisor teeth, eye opening and auditive channel the north of Tocantins, dried into a ventilated oven at opening, and the following reflexology parameters 55ºC, percolated in ethanol and rotary evaporated to were assessed: palmar grasp reflex, auditory startle obtain the crude extract. The extract was partitioned reflex, adult gait, surface righting reflex, and negative with hexane and ethyl acetate to obtain the ethyl geotaxis. Results. There was a greater latency for the acetate extract (EAC). The animals were divided in opening of the eyes in the males and in the females, the two groups (n=16), an experimental group that eruption of the incisor teeth occurred earlier when received EAC in a solution of polysorbate 80 (Tween) compared to the control group. However, the results at 5%, and control group that received only the were within the expected period of development. Also, vehicle. The dose administered of the EAC was 721 male pups presented higher latency in the surface mg.Kg-1 body weight, with an average exposure to righting reflex on postnatal days (PND) 3, 6, 7 and 8.55 mg.kg-1 of flavonoids, to simulate the natural females in PND 5. The other parameters did not differ conditions of cattle intoxication. Female rats were significantly between control and experimental groups. treated for 30 days prior to mating, during the The results could be attributed to the alkaloids cohabitation period and during the gestational period quinoline of P. colorata leaves, for its inhibitory action (GD20), in agreement with the protocols of EPA on GABA3, which is important for cerebral 19961. During the experimental period daily food and maturation. Conclusion: Consumption of EAL during water consumption and body weight were monitored. gestation and lactation promoted a delay of motor On GD21 the animals were euthanized and for each development in the offspring of rats. fetus, the placenta weight was recorded and histomorphometric examination was performed. Financial support / Acknowledgments: References: Results: Phytochemical analysis of the EAC revealed 1. U.S. Environ. Protec. Ag. (1996) Health Effects Test the presence of flavonoids and the quantification of Guidelines OPPTS 870.3800: Reproduc. and Fertility total flavonoids was 4.53%. The experimental group Effects (Draft). Fed. Reg. 61(212):56274-56322. 2. showed an increase in feed intake before and during Organiz. for Econ. Coop. and Develop. (2007) First pregnancy, and an increase in placental weight and in addendum to OECD guideline 426 for testing of giant trophoblastic and spongiotrophoblastic cells in chemicals, “Developmental Neurotoxicity Study.” the basal zone and in the trophoblasts in the labyrinth OECD, Paris, pp. 1-26. 3. Chebib, M.; Duke, R. K.; zone related to control group. It is known that some Duke, C. C.; Konnor, M.; Mewett, K. N.; Johnston, G. flavonoids can induce estrogenic or antiestrogenic A. R. Convulsant actions of calycanthine. Toxicology effects in E2-target cells2. Thus, the active compounds and Applied Pharmacology, 190, p. 58-64, 2003. of P. colorata may have promoted an antiestrogenic Acknowledgments: Work supported with grants from activity in the experimental rats. Conclusion: The CAPES (Process: 23038.007219/2011-79 PNPD results demonstrate the presence of flavonoids in the 2011). ethyl acetate extract which may have caused the toxicity in female rats and the deleterious effects on the placenta. EX 35 - PSYCHOTRIA COLORATA ETHYL ACETATE EXTRACT PRODUCES PLACENTAL Financial support / Acknowledgments: References: ALTERATION 1. U.S. Environ. Protec. Ag. (1996) Health Effects Test Guidelines OPPTS 870.3800: Reproduc. and Fertility JAVAÉ, N. R. K. 1; MOURA, L. T. S. 1; Effects (Draft). Fed. Reg. 61(212):56274-56322. 2. VASCONCELOS, I. C. 1; MARIANO-SOUZA, D.P. Feng, Y.; Zhang, P.; Zhang, Z.; Shi, J.; Jiao, Z.; Shao, 2; PAIVA, J. P. 3; MARUO, V. M. 1. B. Endocrine disrupting effects of triclosan on the placenta in pregnant rats. Plos One, v. 11, (5), 2016. 1. School of Veterinary Medicine and Animal Acknowledgments: Work supported with grants from Sciences, Federal University of Tocantins, Araguaína - CAPES (Process: 23038.007219/2011-79 PNPD TO, Brazil; 2. Faculty of Biology, Federal University 2011). of Tocantins, Araguaína - TO, Brazil; 3. Faculty of Chemistry, Federal University of Tocantins, Araguaína - TO, Brazil.

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EX 36 - EVALUATION OF IN VITRO TOXICITY Financial support / Acknowledgments: FAPEMIG, OF STANDARDIZED FRACTION OF COSTUS CNPq, CAPES and UFMG. SPIRALIS AND ITS POTENTIAL OF REDUCING NEPHROTOXICITY INDUCED BY CISPLATIN IN RATS EX 37 - ELABORATION OF A COMPARATIVE EMBRYOLOGY ATLAS AS A TOOL FOR BOTH SOUZA, L. C. R. 1; AMORIM, J.M. 2; FREIRE, G. P. TEACHING AND RESEARCH IN 1; SOUZA, R. A. L. 2; CASTILHO, R.O. 2; DEVELOPMENTAL BIOLOGY AND TAGLIATI, C.A. 1. TOXICOLOGY

1 LabTox, Universidade Federal de Minas Gerais, VALENCISE, L. 1; DEUS JUNIOR, R.S 1,2; Belo Horizonte – MG, Brasil ; 2 Laboratório de KEMPINAS, W.G. 1. Fitoquímica, Universidade Federal de Minas Gerais, Belo Horizonte – MG, Brasil. 1. Department of Morphology, Institute of Biosciences of Botucatu, UNESP. Botucatu - SP, Brazil. 2. Institute Introduction: Costus spiralis is traditionally used in of Biology, University of Campinas. Campinas - SP. Brazil for the treatment of inflammatory and urinary disorders, particularly for expelling kidney stones, but Introduction and objective: The study of animal there are no studies on its toxicity and prenatal development is fundamental, especially in nephroprotective potential. Renal stone is a common teaching and in research applied to Developmental disease occurring in 8% of the population and is Biology and Toxicology. To fully understand possible associated with the high mortality rate. In addition the deleterious effects caused by toxicants in embryos, it is incidence of acute renal failure (ARF) has been necessary to know what is considered normal prenatal increased in recent years. Considering its potential and development and the implications of such alterations in the need for new drugs for the treatment of ARF, a the individual’s adult and reproductive life. standardized fraction (EHE80) was developed and a Furthermore, knowledge about comparative model for nephrotoxicity assessment was designed. embryology is required to interpret and correlate Objective: Evaluation of the in vitro toxicity and results obtained from experimental models to human potential of reducing nephrotoxic effects of cisplatin in development and fertility. Thus, the purpose of this rats of EHE80. Material and Methods: Cytotoxicity work was to gather information regarding prenatal was assessed using Hek-293 (Human Embryonic development in the form of a Comparative Kidney) and HepG2 (human hepatocellular carcinoma) Embryology Atlas to be used by students and cells, with 24h MTT exposure. The concentrations researchers that study Developmental Biology and used were 0, 15, 20, 30, 100, 150, 200, 300 μg/mL Toxicology. Methods: Embryos and fetuses used in (Hek-293) and 0, 50, 100, 150, 200, 300, 400, 600 previous research projects conducted at the Laboratory μg/mL (HepG2). The absorbance of each well was of Reproductive and Developmental Biology and measured at 570 nm in the ELISA reader. In the in Toxicology were colected and slides were made from vivo experiment Wistar rats were used in pre, post and them. In addition, these slides and also the slides pre+post cisplatin injury (7,5mg/kg) in three dosages collection of the Institute of Biosciences of Botucatu (5, 15, and 30 mg/kg) to evaluate the dose-response (IBB) were scanned and analyzed with Pannoramic dependence. The animals received EHE80 by gavage Viewer software. The images gathered from IBB’s once a day, according to the established model, with slides collection were used to form part of the the maximum time of 10 days. Urine was collected Comparative Embryology Atlas, as guides to evaluate daily and, at the end of the experiment, blood, liver normal prenatal development and also in Embryology and kidneys were collected for further analysis. classes. Results: Based on the information collected to Results: In the cytotoxicity assay, IC50 of 100 μg/ml form the Comparative Embryology Atlas, it was (HepG2) and 150 μg/ml (Hek-293) were found. In the possible to recognize malformations in some of the in vivo model, EHE80 showed a reducing effect of fetuses acquired from previous research conducted at nephrotoxicity. Our results suggest post-treatment the Laboratory of Reproductive and Developmental could be an option to treat nephrotoxicity caused by Biology and Toxicology. In addition, the material cisplatin. All groups presented plasma creatinine produced was used in Embryology teaching by values without difference of the negative control IBB/UNESP students, who approved the content. group, indicating that there was recovery of renal Conclusions: The Atlas of Comparative Embryology damage to a condition close to basal. Conclusion: may be a useful tool for evaluating disturbs in prenatal EHE80 proved to be a good candidate for a new drug. development caused by chance or by the effect of However, further studies are needed with other toxicants. Moreover, it is possible to glimpse a experimental models. Keywords: Costus spiralis, translational application, as well as in teaching and Nephrotoxicity, in vivo, in vitro. research of Developmental Biology and Toxicology. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 113

Financial support / Acknowledgments: FAPESP accumulation of reactive species, culminating in 2015/26395-7 apoptotic cell death, demonstrated by the maintenance of phosphatidylserine on the external face of the membrane cellular, by condensation and nuclear EX 38 - MITOCHONDRIAL DAMAGE PLAYS A fragmentation, add to presence of pro-apoptotic factors KEY ROLE IN HEPATOTOXICITY MEDIATED in cytosol, such as cytochrome-C and AIF besides the BY APOPTOSIS IN HEPATOBLASTOMA activation of caspases 3 and 9. Ours results together CELLS EXPOSED TO FLAME RETARDANTS, demonstrate that BDE-153 lead to cell death by BDE-153 apoptosis with mitochondrial pathways, reinforcing the idea that this class of compound represents a risk to the PEREIRA, L. C.; MIRANDA, L.F.C3; FRANCO- exposed population. Key Words: Mitochondrial swell; BERNARDES, M.F1; TASSO, M.J.3;DUARTE, HepG2 cell; Cell death; hexaBDE-153. F.V4; VARELA, A.T.I4; ROLO, A.P4; PALMEIRA, Financial support / Acknowledgments: São Paulo C.M.M4; DORTA, D.J3. Research Foundation – FAPESP (Proc. 2015/15742-8), the National Council for Technological and Scientific 1 Department of Clinical, Toxicological and Development (CNPq), and Coordination for the Bromatological Analysis, Faculty of Pharmaceutical Improvement of Higher Education Personnel (CAPES) Sciences of Ribeirão Preto, University of São Paulo, for financial support (process number PVE- Ribeirão Preto, Brazil. 2. Department of Bioprocesses A018/2012). and Biotechnology, Faculty of Agronomic Sciences of Botucatu, São Paulo State University, Botucatu, SP. 3Departamento de Química, Faculdade de Filosofia, EX 39 - MITOCHONDRIAL ELECTRON Ciências e Letras de Ribeirão Preto, Universidade de TRANSPORT CHAIN IS A TARGET TO São Paulo, Brasil. 4Department of Life Sciences, TRICLOPYR EXPOSURE Faculty of Sciences and Technology, University of Coimbra, Coimbra, Portugal. PEREIRA, L. C.; BRANDÃO, T.A1; DA SILVA, A.C.G1; ALVES, A.A1; DE OLIVEIRA, B2; Polybrominated diphenyl ethers (PBDEs) are used as DORTA, D.J3 flame retardants in polymer-containing products and, more precisely the BDE-153 congener is one of the 1 Biomedical Sciences Graduate Program, Centro most present in human samples and animal wildlife. Universitário Hermínio Ometto, UNIARARAS, This fact had raised concerns about their toxicity. Araras, SP. 2Department of Clinical, Toxicological However, the mechanism of action of this particular and Bromatological Analysis, Faculty of compound is not well described. This study Pharmaceutical Sciences of Ribeirão Preto, University characterizes the in vitro toxicity the BDE-153, in an of São Paulo, Ribeirão Preto, Brazil. 3Department of attempt to elucidate their mechanism based on Chemistry, Faculty of Philosophy, Sciences and mitochondrial dysfunction and cellular injury using Letters of Ribeirão Preto, University of São Paulo, isolated liver mitochondria and HepG2 cells exposed Ribeirão Preto, SP. 4Department of Bioprocesses and to BDE-153 (0,1 – 50 µM). Mitochondrial parameters, Biotechnology, Faculty of Agronomic Sciences of such as respiration; mitochondrial membrane potential Botucatu, São Paulo State University, Botucatu, SP. (Δψ); swelling; interaction with membrane; Ca+2 release, ATP levels and also redox state were Emerging Contaminants include a long list of routinely evaluated after exposure. Furthermore, the effects of employed products such as pesticides, with reported BDE-153 in HepG2 cells were evaluated by different presence in environment and that presenting a real or endpoints such as, proliferation and 3-(4,5- potential risk to human health or the environment and dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium lack of data on their toxicity and lack of legislation to bromide (MTT) reduction capacity; exposure of control their use. In this context, the herbicide triclopyr phosphatidylserine to the outer cell membrane, nuclear has recently been classified as an emerging condensation and fragmentation, and quantification of contaminant. Data in the literature describe it with pro-apoptotic factors in mitochondrial and cytosolic moderate toxicity in rats and high in humans and fractions after 24 and 48 hours to exposure. Significant aquatic organisms. However, its mechanism of toxicity results demonstrate the ability of BDE-153 to cause is not well described. This study characterizes the in mitochondrial dysfunction, by cyclosporin-sensitive vitro mitochondrial toxicity the triclopyr, in an attempt mitochondrial swelling. In addition to the ability to to elucidate their mechanism using isolated liver decrease the reduction of MTT, proliferation and cell mitochondria exposed to triclopyr (1 – 100 µM). cycle interferences. These cytotoxicity of BDE-153 are Briefly, the parameters of mitochondrial respiration related to a mitochondrial dysfunction, since BDE-153 were assessed polarographically according Chance & generates dissipation of Δψ, formation and Willians (1956). Mitochondrial membrane potential ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 114

(Δψ) was analyzed using safranine-ο as a probe. The evaluate the effects of honey and its major phenolic ability of the compound to interfere with mitochondrial compounds on changes related to complex phenotypes permeability was monitored by the swelling assay and (erected wing posture) and mitochondrial aspects redox state was evaluated by oxidation of SH proteins induced by hypoxia/reperfusion in flies. Flies were after triclopyr exposure. Significant results pre-treated for 3 days with 10% honey and different demonstrate the ability of triclopyr to cause concentrations of caffeic acid and p-coumaric acid. mitochondrial dysfunction, by interference of the Then, submitted to hypoxia for 3 hours followed by herbicide on bioenergetic parameters, causing reperfusion. As results, some flies exposed to inhibition of oxidative phosphorylation, observed by hypoxia/reperfusion acquired a permanent erected decrease in RCR with dose-dependent and increase in wings posture and this aspect was closely correlated the ADP/O rate at the highest concentration assessed with mortality. Differently from honey, caffeic and p- (100 µM). Although the exposure to triclopyr has not coumaric acids displayed no protection against erected been able to cause swelling in organelles and also did wings and locomotor damage caused by not induce the oxidation of the mitochondrial hypoxia/reperfusion. Honey treatment decreased the membrane thiol groups, ours results demonstrate that frequency of flies with this complex phenotype. This the presence of triclopyr was able to cause a small phenomenon is being reported for the first time, and decrease in the Δψ, also at the higher concentration could be associated with down regulation of the Erect (100 μM). Our results demonstrate the toxic potential Wing (EWG) gene found in flies exposed to hypoxic of triclopyr herbicide through mitochondrial conditions. EWG is a transcription factor and in flies dysfunction. Knowing the importance of mitochondria has an important function in the neuronal and muscular to cells, the observed effects can culminate in cell development. In humans, the EWG homologue (NRF- death, reinforcing the idea that this class of compound 1) regulates the expression of genes necessary for represents a risk to the exposed population. Key mitochondrial respiratory function and mitochondrial Words: Mitochondrial damage; Emergent biogenesis. In addition, we results showed that Contaminants; Triclopyr. hypoxia/reperfusion drastically reduced the O2 fluxes in flies mitochondria and honey protected against Financial support / Acknowledgments: Centro decrease of the Opa1 gene expression that is required Universitário Hermínio Ometto, UNIARARAS, for mitochondrial fusion. Our data contributes in a Araras, SP. pioneering way on change in the flies’ wings posture phenotype when exposed to hypoxia/reperfusion situation. As well as, we have shown a potential EX 40 - HONEY PROTECTS AGAINST WINGS positive effect of a powerful antioxidant food against POSTURE ERROR AND MOLECULAR these changes. CHANGES RELATED TO MITOCHONDRIAL PATHWAYS INDUCED BY Financial support / Acknowledgments: CNPq, HYPOXIA/REPERFUSION IN ADULT FAPERGS and CAPES DROSOPHILA MELANOGASTER

CRUZ, L. C. 1, 2, ECKER, A. 1, RODRIDRES, N. 2, EX 41 - CELLULAR EFFECTS OF THIRDHAND MARTINS, I. K. 2, POSSER, T. 2, BARBOSA, N. V. TOBACCO SMOKE FROM SMOKERS' HOMES 1, FRANCO, J. L. 2 FIGUEIRÓ, L. R. 1; LINDEN, R. 2; ZIULKOSKI, A. 1. Department of Biochemistry and Molecular L. 2; DANTAS, D. C. M. 1. Biology, Universidade Federal de Santa Maria-RS, Brazil; 2. Interdisciplinary Center for Biotechnology 1 Federal University of Health Sciences of Porto Research, CIPBIOTEC, Universidade Federal do Alegre (UFCSPA), Porto Alegre, RS, Brazil; 2 Feevale Pampa, Campus São Gabriel- RS, Brazil. University, Novo Hamburgo, RS, Brazil.

Honey is a complex product produced by honeybees Introduction: After the cigarette is extinguished, many from nectar, being widely used for its sweetening toxic compounds remain in the environment and properties, as well as for its benefits to human health. accumulate in the air or on surfaces. This exposure is Hypoxia followed by reperfusion is a condition termed Thirdhand Smoke (THS) and its risks are triggered in several pathologies and can culminate with poorly known. Aim: To evaluate the cellular effects of oxidative tissue damage. In a previous study, we have THS from smokers’ homes. Materials and Methods: demonstrated the protective effect of Brazilian Pampa Cellulose papers (25cm2) were placed in nine biome honey against pro-oxidative effects of smoker’s home and three nonsmoker’s homes. An area hypoxia/reperfusion in adult Drosophila melanogaster. equivalent to the paper size was cleaned with cotton In this study, we conducted an investigation to wipe. These samples were immersed in Dulbecco's ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 115

Modified Eagle's medium (DMEM+THS) at a ratio of The product is considered toxic when it kills in 50 mg material to 2 mL culture medium. A549, Hep-2 concentrations ≤ 30 μg/mL. For acute toxicity the and 3T3 cells were exposed to DMEM+THS for 24 extract (250 mg/kg) was administered by gavage to 6 hours and cellular functions were assessed by males and 6 females per group while the control group tetrazolium dye reduction (MTT), Neutral Red received vehicle alone. The behavioral changes and reuptake (NR) and Trypan blue exclusion assays. signs of toxicity and/or death were observed Results: Smokers' homes showed to be a potential continuously for 1 h after the administration, reservoir of THS pollutants. Concentration of nicotine intermittently for 4 h and over a period of 24 h. The in these homes ranged from bellow the limit of mice were further observed once a day, to 14 days. The detection (0.5) to 759.9 ng/cm2. Cellular proliferation consumption of water and feed as well as body weight was similar in almost all samples after THS exposure. were measured. On day 14, peripheral blood samples Changes in the cellular functions were observed, from the controls and treated mice were collected and mainly higher mitochondrial activity, in paper samples. the levels of urea, creatinine, alanine aminotransferase Moreover, there was positive correlation between and aspartate aminotransferase were determined. The nicotine concentration and mitochondrial viability of hemoglobin level, erythrocyte count, hematocrit, red Hep-2 cells to both paper (p=0.006) and cotton cell index such as mean corpuscular volum, mean samples (p<0.001). In 3T3 cells, there was a negative corpuscular hemoglobin and mean corpuscular correlation between nicotine concentration and hemoglobin concentration were determinated. The lysosomal viability to paper samples (p<0.001) and a animals were euthanized the kidneys and liver were positive correlation between nicotine concentration excised for determination organ weight/body weight. and mitochondrial viability to cotton samples Histopathological studies were carried out to detect (p=0.048). Conclusion: Even showing no cytotoxic possible signs of toxicity. Results: Propolis at effect, the THS was able to cause changes in important concentrations up to 500 μg/mL did not induce death functions for suitable cell survival, for instance, in A. salina. Hyperactivity was observed shortly after increased mitochondrial activity. The results administration of propolis in mice, but no death demonstrated MTT, NR and Trypan assays may be occurred. There were no changes in water and feed performed in A549, Hep-2 and 3T3 cells to evaluate intake, body weight or biochemical and hematological biological effects at the cellular level of THS exposure. parameters. A mild parenchymal inflammatory This is the first study to evaluate the cellular effects of infiltrate has been observed in the kidneys that may THS samples collected from smokers’ homes. indicate mild to moderate dose-dependent toxicity. Conclusion: Red propolis presented low toxicity to A. Financial support / Acknowledgments: Feevale, salina and slight acute toxicity for M. musculus. UFCSPA Financial support / Acknowledgments: CAPES / CNPq EX 42 - INVESTIGATION OF THE TOXICITY OF THE RED PROPOLIS EXTRACT FROM PARAÍBA, BRAZIL EX 43 - EVALUATION OF THE OXIDANT, ANTIOXIDANT AND CYTOTOXIC ACTIVITY PONTES, M. L. C. 1; LIRA, A. B. 1; OLIVEIRA, K. OF THE HYDROALCOLIC EXTRACT OF RED M. O. 1; PACHECO FILHO, E. F. 1; FARIAS, I. A. P. PROPOLIS OF PARAÍBA, BRAZIL 1; SAMPAIO, F. C. 1; PAZ, A. R. 1; LIMA, C. M. B. L. 1; DINIZ, M. F. F. M. 1; PESSÔA, H. L. F. 1. PONTES, M. L. C. 1; GONÇALVES, G. F. 1; LIRA, A. B. 1; PEDROSA, Z. V. 1; DINIZ, M. F. F. M. 1; 1. Universidade Federal da Paraíba, João Pessoa - PB, PESSÔA, H. L. F. 1. Brasil. 1. Universidade Federal da Paraíba, João Pessoa - PB, Introduction: Red propolis is produced in the Brasil mangrove region of northeastern Brazil and has antimicrobial, anti-inflammatory and antifungal Introduction: Red propolis is produced by bees from effects, but little is known about its toxicological the resinous exudate of Dalbergia ecastophyllum (L) effects. Objective: To investigate the toxicity of red Taub. Several biological activities have already been propolis to Artemia salina (A. salina) and acute described for red propolis but little is known about its toxicity in Mus musculus (Swiss). Materials and toxicity. Objective: The present study aimed to Methods: A. salina eggs were hatched in artificial sea evaluate the oxidant, antioxidant and cytotoxic activity water with aeration at 28 ° C for 48 hours. Then the of red propolis. Materials and Methods: Samples of red nauplii were exposed to different concentrations of propolis were obtained from colmeias located on the propolis (125, 250, 500, 1000 μg/mL) for 24 hours. coast of Paraiba (Brazil) and solubilized in ethanol ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 116

70% (10 mg/mL). Samples of 1.5 mL of whole blood Examine the in vivo estrogenic and antiestrogenic was mixed with 4.5 mL of NaCl 0.9%, centrifuged at effects of dipyrone in the uterotrophic assay. Methods: 3.000 rpm for 5 min and the supernatant was removed Immature (22-days-old) rats were treated for three by gentle aspiration. This process was repeated two days with different doses of dipyrone (50, 100, 200 more times. For evaluation of oxidant activity the mg/kg/day) combined with oily vehicle (canola oil) or erythrocytes were resuspended in PBS supplemented estradiol (1 and 10 μg/kg/day). Dipyrone was with glucose (200 mg/dL) to a final hematocrit of 35%. dissolved in distilled water and estradiol in canola oil. The erythrocytes were incubated with propolis (10, 50, All treatments were performed daily by oral gavage. 100, 250, 500 e 1000 μg/mL) at 100 rpm, 22±2°C for Following 24 h of the last treatment, uteri were 60 min. The erythrocyte suspension was the negative carefully removed and dissected, and the full and control and the erythrocyte suspension plus empty uterine weights were recorded. Results: phenylhydrazine (1 mmol/L) was the positive control. Animals that received estradiol at 1 and 10 μg/kg/day Afterwards the samples were aerated and exposed to showed an increase in absolute uterine weight when phenylhydrazine (1 mmol/L) for a further 20 min compared to vehicle-only rats. Dipyrone administered under the same conditions. The methaemoglobin (630 to oily vehicle-treated rats did not increase uterine nm) concentration as a percentage of the total weight at any dose tested, indicating absence of hemoglobin (540 nm) was measured to evaluate the estrogenic activity. However, coadministration of oxidant and antioxidant effect. For evaluation of dipyrone and estradiol (1 and 10 μg/kg/day) cytotoxicity the same samples of propolis was added to significantly decreased uterine weight at all three the erythrocytes suspension (0,5%) in NaCl 0.9%, dipyrone doses in comparison to the group treated with incubated at 100 rpm at 22±2°C for 60 min and estradiol and water. The results were similar for full centrifuged at 3,000 rpm for 5 min. The absorbance of and empty uteri and after correction for body weight. the supernatants was determined (540 nm) to measure Conclusion: These results suggest that dipyrone the extent of erythrocyte lysis. A suspension of produces antiestrogenic effects trough blockage of erythrocytes was used as a negative control (0% estradiol activity on uterine weight in vivo. Additional hemolysis), and the erythrocyte suspension plus 1% studies are needed to confirm these data and elucidate Triton X-100 (100% hemolysis) was the positive the underlying mechanisms. Also, future studies control. All the experiments were performed in should focus on early and late consequences of triplicate. Results: Propolis did not promote developmental dipyrone exposures, with particular hemoglobin oxidation as well as did not inhibit emphasis on endocrine and reproductive sensitive phenylhydrazine-induced oxidation. Propolis induced endpoints. weak hemolysis (< 45%) to 500 μg/mL in all erythrocytes. Conclusion: Red propolis of Paraiba does Financial support / Acknowledgments: Coordenação not induce oxidative stress nor cytotoxicity. de Aperfeiçoamento de Pessoal de Nível Superior (CAPES – Brasil); Conselho Nacional de Financial support / Acknowledgments: CAPES / Desenvolvimento Científico e Tecnológico (CNPq – CNPq Brasil).

EX 44 - SCREENING FOR IN VIVO (ANTI) EX 45 - ASSESSMENT OF THE ANALGESIC ESTROGENIC ACTIVITY OF DIPYRONE DIPYRONE AS A POSSIBLE (ANTI) ANDROGENIC ENDOCRINE DISRUPTOR SILVA, M. T. P. P.; PASSONI, M. T. 1; PALU, G. 1; AMARAL B. A. S. 1; GRECHI N. 1; ARAÚJO G. 1; SILVA, M. T. P. P.; PASSONI, M. T. 1; MARTINS L. F. 1; BOARETO A. C. 1; GOLART J. KRISTENSEN M. N. 2; WOITKOWIAK C. 3; 1; DALSENTER P. R. 1; MORAIS R. N. 1; MORAIS R. N. 1; BOARETO A. C. 1; AMARAL B. MARTINO-ANDRADE A. J. 1 A. S. 1; GRECHI N. 1; MARTINS L. F. 1; DALSENTER P. R. 1; MUNKBOEL C. H. 2; 1 Universidade Federal do Paraná, Curitiba - PR, STYRISHAVE B. 2; KRISTENSEN D. M. 2; GOMES Brasil C. 3; VAN RAVENZWAAY B. 3; MARTINO- ANDRADE A. J. 1. Introduction: The analgesic dipyrone is widely used in many countries and particularly in Brazil, where it is 1 Universidade Federal do Paraná, Curitiba - PR, ranked as the number one analgesic drug in sales. Brasil; 2 Universidade de Copenhagen, Copenhagen, Despite recent reports on the endocrine disrupting Dinamarca; 3 BASF, Ludwigshafen, Alemanha. activities of several analgesics, like paracetamol, ibuprofen and acetylsalicylic acid, there are no data on Introduction: Mild analgesics are among the most used the potential endocrine effects of dipyrone. Objectives: drugs worldwide and have been recently associated ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 117 with antiandrogenic effects. Regardless of the high EX 46 - EFFECT OF THE MYRCIARIA prevalence of dipyrone use in many countries, its CAULIFLORA PEEL RESIDUE THE potential antiandrogenic effects have not yet been CATALASE ACTIVITY OF RATS TREATED investigated. Objectives: Examine the effects of WITH HYPERLIPIDIC DIET dipyrone on steroidogenesis and receptor-mediated androgenic and antiandrogenic activities in vitro and in LIMA JÚNIOR, C.Q.1; SANTIAGO, G. M. 1; vivo. Methods: For assessment of dipyrone effects on SILVA, M.A.C. 2; SALAZAR, V.C.R. 1. steroidogenesis we measured the production of steroid hormones in human H295R cells after exposure to 1. Pontifícia Universidade Católica de Goiás, Goiânia - dipyrone and two metabolites, 4- GO, Brasil; Universidade Federal de Goiás, Goiânia – Methylaminoantipyrine (MAA) and 4- GO, Brasil. Aminoantipyrine (AA), as well as fetal testicular testosterone production in rats following maternal Introduction: Nowadays, obesity represents a major dipyrone exposure. Androgen agonistic/antagonistic public health problem. It is known that there is a effects were examined in vitro for dipyrone and its two relationship between obesity and increased risk for metabolites in a yeast-based transcriptional activation several related diseases, such as hypertension, reporter gene assay (Yeast Androgen Screen – YAS), cardiovascular problems, type 2 diabetes mellitus, and in vivo for dipyrone through the Hershberger dyslipidemias, immunological changes, and oxidative assay, a screening test that uses androgen-dependent stress (OE). OE may cause lipoperoxidation of cell tissues as biomarkers of androgenic action in castrated membrane, aggressions to tissue proteins, enzymes and rats. In vitro we tested a wide range of concentrations DNA. Antioxidant enzymes and antioxidant of dipyrone, MAA, and AA (0.1-1000 M) while in compounds present in several plant extracts may the in vivo assays we used three human relevant control those damages Plants such as Myrciaria dipyrone doses (50, 100, 200 mg/kg/day). Results: In cauliflora has important antioxidant compounds that the H295R steroidogenesis assay, dipyrone, MAA and may assist antioxidant enzymes activity. Objective: To AA reduced the production of androgens and evaluate the effect of the peel residue extract of corticosteroids. For MAA and AA, the lowest Myrciaria cauliflora bark on catalase enzyme of rats concentration to reduce testosterone levels (314 M) treated with hyperlipidic diet. Methods: There were was approximately 4 and 13 times higher than the used 46 Wistar rats. They were divided into two maximum plasma concentrations reported in humans, experiments with different treatment times, 8 and 12 respectively. However, no effects were observed in the weeks. For each experiment rats were subdivided into fetal testicular testosterone production assay at any 4 groups that were treated with different diets: Diet dose. In the YAS and Hershberger assays no androgen with standard ration (Ct), standard ration plus extract agonistic/antagonistic activities were observed. of Myrciaria cauliflora (Ct + Ex), Hyperlipid diet (Hp) Conclusion: In the tested experimental conditions, and Hyperlipid diet plus extract (Hp + Ex). After that, dipyrone and its metabolites can inhibit blood was collected from them and the activity of the steroidogenesis in vitro, but appear not to do so in enzyme catalase was evaluated. Results: There was no vivo. Receptor-mediated androgenic/antiandrogenic statistically significant difference in the activity of the effects were not observed in vivo or in vitro. Thus, enzyme catalase between the groups of animals treated dipyrone has the potential to inhibit steroidogenesis, for 8 weeks, but there was a trend of increased activity however only at concentrations that are not relevant of the enzyme in the Ct + Ex and Hp + Ex groups (p = under normal medical use. 0.0535) and between the group Ct and Hp (p = 0.0568). Regarding the second experiment, a Financial support / Acknowledgments: Coordenação statistically significant difference was observed in the de Aperfeiçoamento de Pessoal de Nível Superior activity of the catalase enzyme between the groups of (CAPES – Brasil); Conselho Nacional de animals treated with the control diet Ct and the group Desenvolvimento Científico e Tecnológico (CNPq – treated with Hp diet (p ˂0.01). In addition to Brasil). increasing trend between Hp + Ex and Hp (p = 0.0512) was observed. Conclusion: Obesity induced by hyperlipid diet altered the activity of catalase and although extract of peel residue of Myrciaria cauliflora did not show statistically significant results under the activity of the enzyme, there was a tendency to try to reverse the changes caused by the hyperlipid diet.

Financial support / Acknowledgments: Pontifícia Universidade Católica de Goiás

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EX 47 - HEMATOLOGICAL AND EX 48 - EFFECTS OF PRE AND POSTNATAL BIOCHEMICAL REFERENCE PARAMETERS EXPOSURE TO AIR POLLUTION ON OF WISTAR RATS FROM THE BIOTERIUM OF BEHAVIORAL AND INFLAMMATION IN MICE THE PUC-GOIÁS BRAIN

GONÇALVES, F.C. 2; LIMA JÚNIOR, C.Q. 1; DI DOMENICO, M. 1; BENEVENUTO, S. G. M. 1; GOMES, C.M. 2; BLANCH, G.T. 2; COSTA, S.H.N. VERAS, M. M. 1. 2; SILVA, M.A.C. 2; SALAZAR, V.C.R. 2; PENNA, K.G.B. 2. 1 Universidade de São Paulo, São Paulo - SP, Brasil.

1. Pontifícia Universidade Católica de Goiás, Goiânia - Introduction: Air pollution is associated with negative GO, Brasil; 2. Universidade Federal de Goiás, Goiânia effects on health, including neurological and – GO, Brasil. behavioral disorders. Studies suggest that gestational and/or postnatal exposure to air pollution would affect INTRODUCTION: Countries with a long tradition in the development and would induce brain injury. the maintenance of bioterium have determined the Damages in brain have been relating to reference values of their animals in order to evaluate neurodevelopmental disorders. Aim: To investigate if the homeostasis and alterations caused by pathological pre and postnatal exposure to air pollution would processes. These values are now used by bioterium induce behavioral toxicity mediated by changes in worldwide, however biochemical and hematological brain glial activation on murine brain. Material and parameters can vary from one animal to another, so it methods: BALB/c mice were daily exposed to current is necessary for each bioterium to obtain its own levels of PM2,5 from São Paulo city during pre and reference values, since intraspecies variations may postnatal period using the Concentrated Ambient exist. OBJECTIVE: The objective of this study was to Particles (CPA). The animals were distributed determine reference values for Wistar rats from the according to pre/postnatal exposure period in four PUC-Goiás Bioterium. METHODS: This study was treatment groups: control (AF/AF), postnatal exposed approved by CEUA-PUC/GO under number 0017-1. (AF/CPA), gestational exposed (CPA/AF) and There were used 17 Wistar rats with a mean weight of gestational and postnatal exposed (CPA/CPA). In the 344 g, obtained from the laboratory of PUC-Goiás, 90 postnatal day, mice performed behavioral test to blood was collected by cardiac puncture. Blood with assess the impact of gestational and/or postnatal PM2,5 EDTA as anticoagulant was used for the hematological exposure on locomotor activity (open field) and analysis and serum for the biochemical analysis. For memory (novel object recognition). In addition, marker each parameter evaluated, the values were expressed as of glial activation (IBA-1) was assessed in cerebral mean, standard error (SD) or standard deviation cortex. Results: Mice from AF/CPA presented higher (SD)and for the purpose of comparison, some results locomotor activity when compared with AF/AF obtained in the literature were also presented. (P=0.002) and CPA/AF (P=0.014) groups. Also the RESULTS: In the present study, values were found CPA/CPA group presented higher locomotor activity that deviate from the range of values found in other than AF/AF (P<0.001) and CPA/AF (P=0.004) groups. studies when the averages were observed, among Besides this, animals exposed to PM2,5 in postnatal which we have the number of platelets (767,65± period (AF/CPA and CPA/CPA) spent more time 105,8510³/µL), lymphocytes (93,85 ± 1,84%), exploring the objects than mice does not exposed in neutrophils(4,81 ± 1,64%), eosinophils (0,04 ± 0,05%), this period (AF/AF and CPA/AF). Regarding to glial Aspartate Aminotransferase (126,29 ± 4,99 U/L) and activation, the IBA-1 levels were higher in AF/CPA alkaline phosphatase (257,65 ± 11,40 U/L). group compared to AF/AF (P=0.018) and CPA/AF CONCLUSION: Studies in experimental toxicology (P=0.043) groups and it also it was higher in use reference values to prove the intoxication, and CPA/CPA group compared to AF/AF and CPA/AF some parameters evaluated in this study were (P<0.001, respectively) groups. Conclusion: Although discrepant in relation to other studies, highlighting the the mechanisms of these effects need to be elucidated, need for each bioterium to determine its own reference these findings suggest that exposure to air pollution values for homeostasis evaluation, disease mainly in the postnatal period may represent a development and the process treatment and/or significant risk factor for central nervous system prevention in intoxication models during scientific diseases / disorders. experiments. Financial support / Acknowledgments: CAPES. Financial support / Acknowledgments: Pontifícia Universidade Católica de Goiás

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EX 49 - ACUTE ORAL TOXICOLOGICAL EX 50 - REMOVAL OF BOTHROPIC VENOM EVALUATION OF ANTICANCER FROM RAT CIRCULATION: A COMPARATIVE PROTOTYPES DIHYDROPYRIMIDINONES STUDY USING ANTIVENOM AND DERIVATIVES 4P AND 4BC HEMOPERFUSION

MELLO, M. H.; MATOS, L.H.S.1; MAIA, T.F.1; DINI, M. M. J.; YOSHIDA, E. H.1; HANAI, V. M.1; FERREIRA, C.L.; NÓBREGA, Y.K.M.1; CASTRO, GROTTO, D.1; OSHIMA-FRANCO, Y.1; M.B.1; KUCKELHAU, S.A.S.1; CORRÊA, J.R.1. CAVALCANTE, R. L.1; CAMPANHOLI, J.1; OLIVEIRA, M. E.1; MORENO, F. S. 1;ARANHA, E. 1 Universidade de Brasília, Brasília - DF, Brasil. F. C.2;HISLOP, S. 3; DIAS, L. 3; COGO, J. C. 4.

Cancer is a complex and multifactorial disease that 1 Universidade de Sorocaba, São Paulo - SP, Brasil; 2 present a high incidence and mortality worldwide. Universidade de São Paulo - SP, Brasil; 3 Drugs used in conventional therapy have several Universidade de Campinas, São Paulo - SP, Brasil; 4 adverse effects and are associated to resistance Universidade do Vale do Paraíba, São José dos development. There is a need for the development of Campos - SP, Brasil. new, safer and more effective antineoplastic agents. Kinesin Eg5 inhibitor Monastrol, a Introduction: Snakebite envenoming is a neglected dihydropyrimidinone (DHPM) obtained synthetically tropical disease since the accidents result in important by Biginelli’s reaction, presents high antitumoral sequels and deaths, worldwide. In Brazil, Bothrops activity, and led the search for other DHPM genus were responsible by 86.2% of snake accidents derivatives with interesting pharmacological profiles. (2009-2013). The specific treatment against ophidian Compounds 4p and 4bc showed activity against lethality is the serum therapy, but any effort searching adenocarcinoma cell lines and selective cytotoxicity to minimize the toxic systemic and local effects for tumor cells in previous studies. The present study resulting from snakebites is very important. Objective: aims at the acute oral toxicological evaluation. We compared hemoperfusion to serum therapy against Toxicity was evaluated after oral administration of Bothrops jararacussu venom (Bjssu) injected in rats, both compounds using OECD 423 guide as protocol. under parameters of hematological profile and survival Based on this study for oral exposure the compounds for 24 h. Materials and methods: Wistar rats were were classified as category 5 (LD50 > 2000mg/kg). No divided in 3 groups (n=3, each): I) Hemoperfusion: clinically relevant changes in feed intake, water intake, animals were submitted to a Charcoal/Tannic Acid weight gain and organ index were observed. (CTA) cartridges, made in house, with no venom Histological and anatomopathological studies revealed inoculation. II) Bjssu & Commercial Bothropic normality in all groups. Hematological and Antivenom (CBA): animals envenomed with Bjssu biochemical evaluation showed no difference between (12.5 mg/kg) via intramuscular (i.m.) received CBA control and treated groups. Long-term studies are (via intraperitoneal, i.p.) after 10 min; III) Bjssu & needed to examine whether the low toxicity results will CTA: animals canulated for hemoperfusion were remain low after the continued exposition to these envenomed as Group II, and after 10 min were compounds. The pharmacokinetic profile of the hemoperfused using CTA. Blood samples were compounds and the development of pharmacotechnical collected after 24 h. The study was approved under strategies to improve the pharmaceutical delivery of institutional protocol no. 031/2014. Results and the compounds are the next steps in the development conclusion: The hemoperfusion procedure was of these substances. successful and after 24 h animals had a normal Financial support / Acknowledgments: Fundação de behavior (Group I). The CBA was effective against the Apoio à Pesquisa do Distrito Federal - Processo lethality, but not against the local necrotizing reaction 0193.000961/2015. (Group II). Two animals died during and the third, at the end of hemoperfusion (Group III); at necropsy, animals had hemorrhagic lungs, heart, abdominal cavity and at local inoculation. Hematological results showed a decrease in platelet count (103/µL): 459- 537compared to literature 786-1015, in Group I, with no change on leukocytes or erythrocytes cells. No hematological change was seen in Group II, while no results were obtained in Group III due the deaths of animals. The platelets decrease in healthy animals is considered a secondary effect face to benefits that it could bring in the venom removal. Concluding, strategies to improve the CTA cartridges to avoid the ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 120 effects of envenoming need to be reviewed. The choice control. However, there was a significant increase in of CBA as a positive control was successful. the CAT activity in the CA group in relation to the Financial support / Acknowledgments: Fapesp control. In the kidney, there was a strong trend towards a lower concentration of ROS than in the control. The present study showed that although the water of the EX 51 - EVALUATION OF THE Doce river and the water treated for public supply have BIOACCUMULATION OF METALS AND low concentrations of metals, they are capable of OXIDATIVE STRESS IN RATS TREATED causing changes in the body through the CHRONICALLY WITH WATER FROM THE bioaccumulation and disturbance in the homeostasis of DOCE RIVER AFTER THE FUNDÃO IRON the antioxidant system in the analyzed tissues, MINING DAM confirming that oxidative stress is a common mechanism of metal blending. GUIMARÃES, N. C. 1; NÓBREGA, A. C. F. 2; Financial support / Acknowledgments: PCDF GONÇALVES, M. R. 1; SPIM, S. R. V. 3; GOMES, B. R. B. 4; FERREIRA, E. A. 1; SOUZA, P. E. N. 5 SOUZA, F. H. V. 4; GROTTO, D. 3; SANTOS, V. S. EX 52 - EFFECTS OF IVERMECTIN ON 2. REPRODUCTIVE PARAMETERS IN RABBITS (ORYCTOLAGUS CUNICULUS) 1. Faculdade de Ciências da Saúde, Universidade de Brasília, Brasília-DF, Brasil; 2. Faculdade de MOREIRA, N. 1; TORRES, M. A. 2; GOTARDO, A. Ceilândia, Universidade de Brasília, Brasília-DF, T. 3; ANDRADE, A. F. C. 2; SPINOSA, H. S. 1. Brasil; 3. Laboratório de Pesquisa em Toxicologia, Universidade de Sorocaba, Sorocaba-SP, Brasil; 4. 1. Department of Pathology, School of Veterinary Laboratório de Bioquímica e Química de Proteínas, Medicine and Animal Science, University of São Instituto de Biologia, Universidade de Brasília, Paulo, Brazil; 2. Department of Animal Reproduction, Brasília-DF, Brasil. 5. Laboratório de Ressonância School of Veterinary Medicine and Animal Science, Paramagnética Eletrônica, Núcleo de Física Aplicada, University of São Paulo, São Paulo, Brazil; 3. Center Universidade de Brasília, Brasília-DF, Brasil. for Research in Veterinary Toxicology, Department of Pathology, School of Veterinary Medicine and Animal On November 5th, 2015 the Fundão iron mining dam, Science, University of São Paulo, Brazil. located in Mariana, MG State, collapsed. The objective was to evaluate the toxicity induced by the water of the Introduction and aim: Ivermectin (IVM) is a Doce river after disaster. Rats were chronically macrocyclic lactone used for the treatment of parasitic exposed to the water collected from two points of the infections and widely used in veterinary medicine as river (study I): Naque and Ipatinga, MG (NI) and an endectocide. In mammals, evidence indicates that Regência, ES (RE) and from two cities supplied with IVM interacts with chloride channels mediated by γ- water from the Doce river and with public treatment aminobutyric acid (GABA). The GABAergic system is (study II): Governador Valadares, MG (GV) and involved in the manifestation of sexual behavior. Colatina, ES (CA). In the study I, the blood samples of Previously, we observed that IVM in male rats the NI group showed significant decrease of the GPx decreased testosterone levels without interfering with activity in relation to the Group C. There was an sexual motivation and penile erection of male rats. increase of the CAT of the RE group in relation to the Following these studies and aiming to understand the NI group. The cortex exhibited significant decrease in pharmacological mechanisms involved in the effects of GPx activity in the NI group and RE in relation to the ivermectin in the reproductive sphere, this work control. Although the liver of the RE group had purposes evaluate sperm motility and concentration in significantly lower concentrations of Al and Mn, there rabbits 24 and 48 hours after administration of 0.2 and was a significant increase of the CAT in relation to the 1.0 mg/kg IVM subcutaneously. Material and methods: control group. There was greater bioaccumulation of Male New Zealand rabbits were divided into three Pb and no alteration in the antioxidant system, both in equal groups (N=6). The experimental groups received relation to the control group in the kidney. In study II, 0.2 (therapeutic dose) or 1.0 mg/kg IVM. The control there was a significant increase of GSH in the blood of group received 1.0 ml/kg of saline (0.9% NaCl). It was the CA group in relation to the control. The evaluated the sperm motility and concentration in bioaccumulation of Fe in the cortex the CA group was rabbits 24 and 48 hours after administration of IVM or lower than the Group C. In this tissue, a significant saline. Results and Discussion: No significant effects decrease in GPx activity in the CA group and a strong were observed in sperm motility and concentration at tendency of the GV group were observed, both in 24 and 48 hours after IVM administration. Conclusion: relation to the control. In the liver, the CA group had In conclusion, IVM administration in New Zealand lower concentrations of Fe and Mn in relation to the rabbits did not impair sperm quality, so it was not able ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 121 to modify sperm motility and concentration Intake (ADI) determined by ANVISA is safe regarding parameters. Financial support: FAPESP (Process nº the sexual behavior of male mice treated for a long 2015/03131-4). period.

Financial support / Acknowledgments: FAPESP Financial support / Acknowledgments: (Process nº 2015/03131-4). PIBIC/UEG

EX 53 - EFFECT OF CHRONIC INTOXICATION EX 54 - METABOLIC EFFECTS OF WITH MANCOZEBE ON SEXUAL BEHAVIOR MANGANESE IN THE NEMATODE OF MALE SWISS MICE CAENORHABDITIS ELEGANS THROUGH DAERGIC PATHWAY AND TRANSCRIPTION SILVA, N. O. D. 1; GUIMARÃES, A. J. D. 1; FACTORS ACTIVATION BORGES, K. R. S. 1; COSTA, H. R. F. 1; VERDIONO, W. L. 1; DIAS- JÚNIOR, W. 1; GUBERT, P. 1, 2; PUNTEL, B. 2; LEHMEN, L. 2; MAZARO E COSTA, R. 2. FESSEL, J.P. 3; CHENG, P. 2; BORNHORST, J. 4; AVILA, D.S. 5; ASCHNER, M. 6; SOARES, F. A. A. 1 Universidade Estadual de Goiás, Ceres - GO, Brasil; 2. 2 Universidade Federal de Goiás, Goiânia - GO, Brasil. 1. Universidade Federal do Oeste da Bahia, UFOB, Barreiras-BA, Brazil; 2. Universidade Federal de Santa Introduction: In Brazil, pesticides are accessible, and Maria, UFSM, Santa Maria - RS, Brazil; 3. Vanderbilt their use is expanding, reaching the fourth position in University Medical Center, Nashville - TN, USA; 4. the ranking of countries that use pesticides. Mancozebe University of Potsdam - Nuthetal, DE; 5. Universidade is a widely used fungicide, because it has several Federal do Pampa, UNIPAMPA, Uruguaiana - RS, agricultural and industrial applications, including the Brazil; 6. Albert Einstein College of Medicine, Bronx - control and elimination of about 400 different NY, USA. pathogens, found in vegetables and cereals. Inappropriate use leads to contamination of the Manganese (Mn) is a crucial metal since it acts as a environment and food that reaches the consumer. cofactor for many enzymes, including glutamine Long-term intoxication by Mancozeb can cause synthetase, calmoduline-dependent phosphatase, and endocrine, teratogenic, mutagenic and carcinogenic mitochondrial superoxide dismutase. Nevertheless, the risks. Further studies are needed to clarify the effect on overexposure to Mn has been associated with a male sexual potential, motivation and performance, debilitating neurological syndrome called manganism, aiming to controlling the use of this fungicide. resembling Parkinson’s disease. Furthermore, Mn has Objective: Evaluate the effect of daily and long term been reported to affect lipid metabolism by exposure of Mancozeb on the sexual behavior of male mechanisms which have yet to be established. swiss mice. Methods: Sixty Swiss mice were used, 30 Regarding that, we used the nematode Caenorhabditis males and 30 females, weighing approximately 50g elegans to examine Mn’s effects on the dopaminergic and 50 days old, sexually mature, from the UFG, (DAergic) system and determine which transcription divided into 3 groups: White (no intervention); factors that regulate with lipid metabolism are affected Control: water v.o (gavage); Mancozeb: 0.37 mg / by it. Worms were exposed to Mn for four hours in the KgPC v.o (gavage), treated for 60 days. The dose of presence of bacteria E. coli OP50 and in a liquid the fungicide was calculated based on the acceptable medium (85 mM NaCl). Mn increased fat storage as daily intake (ADI) defined by ANVISA and corrected evidenced both by Oil Red O lipid assays, by the body surface area for mice (Reagan-Shaw et al., concomitantly increasing triglyceride levels. In 2008). After 60 days of treatment, each male was addition, metabolic activity was reduced as a reflection placed with a nulliparous female, in estrus, mature, and of decreased oxygen consumption caused by this sexual behavior recorded on a video camera for 30 metal. Mn also affected feeding behavior as evidenced minutes. Subsequently, video analysis was performed by decreased pharyngeal pumping rate. DAergic to evaluate the male reproductive parameters: neurons viability were not altered by Mn, however the frequency and latency of mounts, intrusions and dopamine levels were significantly reduced following ejaculations, total mounts, copulatory efficiency and Mn exposure. Furthermore, the expression of sbp-1 number of mounts using Test t, with 5% significance. transcription factor and let-363 protein kinase Results: Chronic exposure of mice to mancozeb responsible for lipid accumulation control was fungicide (0.37 mg / KgPC) during 60 days did not increased and decreased, respectively, by Mn. promote changes in any of the parameters of sexual Altogether, our data suggest that Mn increases the fat behavior studied. Conclusion: The Acceptable Daily storage in C. elegans, secondary to DAergic system ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 122 alterations, under the control of SBP-1 and LET-363 diuron group, the 15-week LI was less than the 07-day proteins. LI, despite the occurrence of hyperplastic reaction, suggesting an adaptive process after prolonged Financial support / Acknowledgments: CAPES exposure. Conclusion: Increased cell proliferation in (Coordenação de Aperfeiçoamento de Pessoal de Nível NaS-fed or in diuron-fed animals has been related to Superior), CNPq (Conselho Nacional de abrasion or to cytotoxicity, respectively. The present Desenvolvimento Científico e Tecnológico), study indicates that the urothelium of the kidney pelvis FAPERGS (Fundação de Amparo à Pesquisa do is as susceptible as the urinary bladder to chemical Estado do Rio Grande do Sul), NIH-USA (National agents that induce cell proliferation through different Institutes of Health). modes of action.

Financial support / Acknowledgments: CNPq - EX 55 - CELL PROLIFERATION IN THE PIBIC- 129475/2015-7, TOXICAM. KIDNEY PELVIS OF RATS EXPOSED TO DIURON OR SODIUM SACCHARIN EX 56 - DIURON CARCINOGENICITY STUDY: BENEDITO, O. L. 1; FAVA, R. M. 1; MORPHOLOGICAL CHANGES INDUCED IN NASCIMENTO E PONTES, M. G. 1; DE THE UROTHELIUM OF RATS CAMARGO J. L. V. 1; DE OLIVEIRA, M. L. C. S. 1. FAVA, R. M. 1; SALES, B. C. P. 1; NASCIMENTO 1 Núcleo de Avaliação do Impacto do Meio Ambiente E PONTES, M. G. 1; BENEDITO, O. L. 1; SOUZA, Sobre a Saúde Humana (TOXICAM), Departamento N. P. 1; SANTOS, I. P. 1; VACCARI, C. 1; de Patologia, Faculdade de Medicina de Botucatu - IHLASEH, S. M. 2; DE CAMARGO, J. L. V. 1; DE FMB- UNESP, São Paulo -SP, Brasil. OLIVEIRA, M. L. C. S. 1.

Introduction: Evaluation of cell proliferation is an 1 Núcleo de Avaliação do Impacto do Meio Ambiente important tool for detection of urothelial cytotoxicity Sobre a Saúde Humana (TOXICAM), Departamento induced by chemicals. The urothelial cell carcinoma de Patologia, Faculdade de Medicina de Botucatu - (UCC) is a highly incidental neoplasia that may occur FMB- UNESP, São Paulo -SP, Brasil; 2 BASF, São in different parts of the urinary system. Occupational Paulo - SP, Brasil. and environmental exposures to chemicals are important risk factors related to this disease. High Introduction: The herbicide diuron (3-(3,4- dietary concentrations of sodium saccharin (NaS), an dichlorophenyl)-1,1-dimethylurea) induced urothelial organic non-caloric sweetener, or of diuron, a tumors in the urinary bladder of Wistar rats fed the substituted urea herbicide, induce urinary bladder dietary concentration of 2500ppm in a long-term study. carcinoma in rats exposed for extended periods of The diuron mode of action involves metabolic time. Our group has shown induction of proliferative activation, cytotoxicity, cell degeneration, necrosis, changes in the urinary bladder urothelium of rats exfoliation, regenerative cell proliferation and, exposed to these compounds for seven days, or 15 and eventually, tumors. Although the role of high dietary 20 weeks. However, proliferative changes induced by concentrations (1250ppm, 2500ppm) of diuron in these compounds in the kidney pelvis urothelium have inducing urothelial lesions is well established, not been systematically described. Objective: To successive studies from our laboratory suggest that the evaluate urothelial morphology and the cell lower concentration of 500ppm may also be cytotoxic proliferation rate in the kidney pelvis of male Wistar and potentialy carcinogenic to the rat urothelium. rats fed 710ppm NaS or diuron 2500ppm for short or Evidence of urothelial cytotoxicity for this relatively long-term exposures. Material and Methods: concentration level may directly impact the risk Tissues analyzed in this study were obtained from a assessment of this substance, since it is five folds previous experiment (Fava et al., 2015) in which forty- lower than that described in the literature as two male Wistar rats were allocated to three groups of carcinogenic (2500ppm) and very close to the 14 animals each that were fed control diet or diet with currently accepted NOAEL of 125ppm. Objective: To NaS 710ppm or diuron 2500ppm. After seven days or verify if long-term dietary 500ppm diuron induces 15 weeks the animals were euthanized; the kidneys urothelial lesions. Material and Methods: One hundred were processed for histological and and thirty two male Wistar rats were allocated to non- immunohistochemical analyses, the PCNA labeling exposed control or to 500ppm diuron-fed groups and index (LI) was used for cell proliferation assessment. evaluated after 20 or 84 weeks. Urinary bladders and Results: Although not significantly, diuron or NaS kidneys were processed for histological analyses. both increased the incidences of simple hyperplasia Results: Diuron-fed animals had survival rates similar (HS) and the LI when compared to the control. In the to the controls, i.e., 66.6% and 69.6%, respectively. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 123

After both experimental periods, no differences were were analyzed by fluorescence microscopy. Statistical observed in water consumption, body weight gain and analysis was performed using ANOVA and Dunnett absolute and relative liver and kidney weights. post-hoc test. RESULTS There were no significant However, the diuron group presented increased results for SRB and MTT assays on 24h of exposure. relative spleen weights and splenomegaly, events After 48h, BDE-209 (25μM) and all BDE-209 + RAP already documented in diuron-fed rats. Food concentrations showed significant decrease for MTT consumption was decreased in the diuron group at the assay. And just BDE-209 + RAP showed significant 20-week time point only, probably due to palatability results for SRB assay. BDE-209 (25μM) was also able adjustment. After 84 weeks, the incidences of simple to stop the cell cycle at G2/M phase in the presence or hyperplasia were significantly increased in the urinary absence of RAP after 48h of exposure. Nuclear bladder (10/18) and kidney pelvis (23/25) of the diuron fragmentation increased for BDE-209 25μM + RAP, group, when compared to the control (2/22 and 9/30). and the number of apoptotic cells increased in Conclusion: Occurrence of urothelial hyperplasia in exposures to both, 10μM and 25μM, with and without rats exposed to 500ppm diuron for 84 weeks suggests RAP. CONCLUSIONS BDE–209 showed cytotoxicity that this concentration is cytotoxic and potentially for HepG2 after 48h of exposure and the induction of carcinogenic to the rat urothelium. autophagy with rapamycin do not prevent the apoptotic Financial support / Acknowledgments: FAPESP - cell death induced by this congener. Proc. 2015/11628-6, CNPq Proc. 140306/2014-5, Financial support / Acknowledgments: CNPq, TOXICAM. CAPES, FAPESP.

EX 57 - THE INDUCTION OF THE EX 58 - NEUROTOXIC EFFECTS OF AUTOPHAGIC PROCESS DO NOT PREVENT AYAHUASCA EXTRACT (BANISTERIOPSIS BDE–209 CYTOTOXICITY TO HepG2 CELLS CAAPI AND PSYCHOTRIA VIRIDIS) IN PREGNANT WISTAR RATS AFTER CHRONIC MIRANDA, R. G. 1; FERRARI, R. S. 1; DORTA, D. EXPOSURE J. 1. JULIANA ALVES DE MORAIS1, LUCIANA 1. Faculdade de Filosofia, Ciências e Letras de GUEIROS MOTTA2, RHAUL OLIVEIRA1, Ribeirão Preto - Universidade de São Paulo, Ribeirão LEONORA MACIEL SOUSA VIANNA3, MARCIA Preto - SP, Brasil. RENATA MORTARI4, RIVADÁVIO FERNANDES BATISTA AMORIM3, ELOISA DUTRA CALDAS2, Polybromamined diphenyl ethers (PBDEs) are ALINE PIC-TAYLOR1. environmental contaminants widely used for fire prevention. Studies showed that PBDEs are found on 1-Laboratory of Embryology and Developmental environmental samples and biological fluids. Biology, Department of Genetics and Morphology, Moreover, PBDE has hepatotoxic potential. Due to Institute of Biological Sciences, University of Brasilia, these facts, there is a big concern with their toxicity. Brasilia-DF, Brazil 2- Laboratory of Toxicology, Since congener BDE-209 has the highest production Faculty of Health Sciences, University of Brasilia, worldwide, the focus of this work was on it. Brasilia-DF, Brazil 3- Department of Pathology, OBJETICVE Evaluate effect of the induced School of Medicine, University of Brasilia, Brasilia- autophagic process on the toxic mechanism of action DF, Brazil 4- Laboratory of Neuropharmacology, of BDE-209 in HepG2 cells. METHODOLOGY Department of Physiological Sciences, Institute of HepG2 was maintained in Minimum Essential Medium Biological Sciences, University of Brasilia, Brasilia- supplemented with 10% of FSB and antibiotic- DF, Brazil. antimycotic 1.2g/L in 5% CO2 atmosphere at 37ºC and treated with BDE–209 (1, 5, 10 and 25μM), with and Ayahuasca is a psychoactive concoction ancestrally without rapamycin 100nM (RAP) for 24 and 48 hours used by indigenous Amazonian populations, and more of exposure. After that, cells were washed and recently consumed in various Christian rituals in Brazil different assays were performed. SRB and MTT assays and other countries. This beverage has shown followed Skehan’s et al. (1990) and Denizot & Lang’s therapeutic potential and requires preclinical studies in (1986) protocols, respectively, and cells were analyzed order to evaluate its safety. Furthermore, non-ritualistic with a spectrophotometry. Cell cycle and Annexin V- use of ayahuasca has increased over the last decade, FITC/PI followed Braylan’s et al. (1982) and thus prompting concern about any potential Zhivotovsky’s et al. (1999) protocols, respectively, and detrimental health effects in this context. This study 10000 events were analyzed by flow cytometry. evaluated the toxicity of ayahuasca in pregnant female Nuclear condensation and fragmentation assay used Wistar rats treated daily from the 6th to 20th Pollack’s et al (1991) protocol and 300 cells per slides gestational days at 1X, 2X, 4X and 8X the ritualistic ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 124 dose (25 animals/dose). A total of twenty-four animals, decrease in viability of 3T3 cells following treatment 44% from the 4X and 52% from the 8X groups, died with EAP (625-2,4 µg/mL) in a concentration- during the experiment after showing clinical signs of dependent manner. The IC50 value obtained was 29,3 toxicity, including: tremors, piloerection, hind limb μg/mL and in vitro estimated DL50 was 363 mg/mL, abduction, cyanoses and convulsion. There was a suggesting that Oral Acute Toxicity test should be statistically significant dose-related neuronal loss, initiated with 2000 mg/kg (Category 5 of toxicity). evaluated by Nissl staining, in CA1, CA2 and CA3 Conclusion: Therefore, it can be concluded that EAP hippocampal regions, together with raphe nuclei, in induced concentration-dependent cytotoxicity in 3T3 comparison to the control group in animals from the cells and in vitro DL50 estimative is useful to reduce two highest doses. Significant macroscopic changes use of animals in toxicological evaluation of this were observed in both the stomach and intestine of the material. surviving animals from all study groups, in addition to Financial support / Acknowledgments: FAPEG - significant histological alterations in the kidneys of Fundação de Amparo a pesquisa do estado de Goiás. animals treated at the two highest doses. These are important preclinical results when considering pharmacological use of ayahuasca and an alert to EX 60 - TOXICOPHORIC AND METABOLIC IN potential abusive use of this beverage. SILICO EVALUATION OF HIBALACTONE

Financial support / Acknowledgments: CAPES; SORAIA FERREIRA BEZERRA; MATHEUS Cnpq, FAPDF. GABRIEL DE OLIVEIRA²; LILIANE DE SOUSA SILVA ² JOSÉ REALINO DE PAULA²; VINICIUS BARRETO DA SILVA³. EX 59 - NEUTRAL RED UPTAKE ASSAY AS A TOOL FOR TOXICOLOGICAL ASSESSMENT ¹Laboratório de Farmacologia e Toxicologia Celular – OF EXTRACT Lafoensia pacari A. ST.-HIL. LFTC, Faculdade de Farmácia, Universidade Estadual (LYTHRACEAE) de Goiás, Goiânia-GO, Brasil; ²Laboratório de Pesquisa de Produtos Naturais - LPPN, Faculdade de GARCIA, S. A. S.; SOUSA, S. A. 1; ANDRADE, W. Farmácia, Universidade Federal de Goiás, Goiânia - M. 2; VALADARES, M. C. 2; BARA, M. T. F. 1. GO, Brasil; ³Escola de Ciências Médicas, Farmacêuticas e Biomédicas, Pontifícia Universidade 1. Laboratório de Pesquisa de Produtos Naturais - Católica de Goiás, Goiânia - Goiás – Brasil. LPPN, Faculdade de Farmácia, Universidade Federal de Goiás, Goiânia - GO, Brasil; 2. Laboratório de Introduction: The physicochemical properties, Farmacologia e Toxicologia Celular - FarmaTec, metabolism and toxicity are relevant parameters in the Faculdade de Farmácia, Universidade Federal de search for new compounds with pharmacological Goiás, Goiânia - GO, Brasil. properties, since they are important features responsible for most drug candidate failures. The Introduction: Lafoensia pacari is a native plant from natural lignan hibalactone have been described with a brazilian cerrado, popularly known as “pacari”, large variety of biological activities, such as “dedaleiro” ou “magava-brava” and used in folk antitumoral, antimicrobian and antioxidant, however medicine as an healing, anti-inflammatory and against there is a lack of studies regarding its metabolism, as gastritis and ulcer. Previous studies have revealed the well as its physicochemical and toxicophoric presence of phenolic compounds, represented by properties. Objective: To investigate the elagitannins in this species. As this species is a physicochemical and toxicity properties of the promising candidate to the development of a novel hibalactone metabolites using computational herbal medicine, toxicological studies are necessary to approaches. Materials and Methods: MetaPrint2D- attest the safety of its use. Objetive: Evaluate the in React online tool was used to predict the metabolic vitro cytotoxic activity of acetone extract obtained pathways of hibalactone with simulations in phase I from L. pacari leaves in basal fibroblasts to estimate and II of potential human biotransformations. The the starting dose for in vivo Oral Acute Toxicity test. analysis of physicochemical properties was performed Materials and Methods: The cytotoxicity assay were with Molsoft DrugLikeness tool and its toxicological performed by Neutral Red Uptake assay in 3T3 cells potentials were investigated using Lazar, Pred-hERG line, according Interagency Coordenation Committee and Pred-Skin tools. Results: The biotransformation on the Validation of Alternative Methods (ICCVAM, analysis with hibalactone revealed at least five possible 2006) protocol. Cells were exposed for 48 hours to metabolites for its molecular structure, concerning decreasing concentrations (625-2,4 µg/mL) of reduction reaction for positions 7 and 8 (metabolite 1), lyophilized acetone extract of L. pacari leaves (EAP) dealkylation reactions for positions 3 and 4 (metabolite before cytotoxicity assessment. Results: There was a 2) and 3' and 4' (metabolite 3), and hydroxylation ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 125 reaction for position 10 and 10' (metabolites 4 and 5, potentials were investigated using Lazar, Pred-hERG respectively). In relation to physicochemical and Pred-Skin tools. Results: The biotransformation properties, the metabolites did not violate the analysis with hibalactone revealed at least five possible Lipinski’s rule of five. The metabolites were not metabolites for its molecular structure, concerning predicted as toxic regarding the Ames toxicity and reduction reaction for positions 7 and 8 (metabolite 1), hERG inhibition. However, a predicted risk of dealkylation reactions for positions 3 and 4 (metabolite carcinogenicity was detected for metabolites 1, 4 and 2) and 3' and 4' (metabolite 3), and hydroxylation 5, and skin sensitization for metabolites 1, 2 and 3. reaction for position 10 and 10' (metabolites 4 and 5, Conclusion: It was possible to propose metabolites for respectively). In relation to pharmacokinetics hibalactone based on its biotransformation reactions, properties, the metabolites did not violate the which were found to have druglike physicochemical Lipinski’s rule of five. The metabolites were not profile consistent with orally active drugs in humans. predicted as toxic regarding the Ames toxicity and Moreover, the predicted toxicities for hibalactone hERG inhibition. However, a predicted risk of metabolites may guide further investigation of their carcinogenicity was detected for metabolites 1, 4 and toxicological mechanisms in biological systems. 5, and skin sensitization for metabolites 1, 2 and 3. Conclusion: It was possible to propose metabolites for Financial support / Acknowledgments: CAPES hibalactone based on its biotransformation reactions, which were found to have druglike pharmacokinetics profile consistent with orally active drugs in humans. EX 61 - EVALUATION OF Moreover, the predicted toxicities for hibalactone PHARMACOKINETICS AND metabolites may guide further investigation of their TOXICOLOGICAL PROPERTIES OF toxicological mechanisms in biological systems. HIBALACTONE METABOLITES BY IN SILICO APPROACHES Financial support / Acknowledgments: CAPES

SORAIA FERREIRA BEZERRA, MATHEUS GABRIEL DE OLIVEIRA²; LILIANE DE SOUSA EX 62 - LOCARSERIN AS A SAFE SILVA ² JOSÉ REALINO DE PAULA²; VINICIUS ALTERNATIVE FOR OBESITY TREATMENT: BARRETO DA SILVA³. DECREASE IN WEIGHT GAIN AND NO DAMAGE TO THE MALE RAT ¹Laboratório de Farmacologia e Toxicologia Celular – REPRODUCTIVE SYSTEM LFTC, Faculdade de Farmácia, Universidade Estadual de Goiás, Goiânia-GO, Brasil; ²Laboratório de PACHECO, T. L. 1; FIGUEIREDO, T. M. 2; Pesquisa de Produtos Naturais - LPPN, Faculdade de JÚNIOR, R. S. D. 1; BORGES, C. S. 2; ROSA, J. L. 2; Farmácia, Universidade Federal de Goiás, Goiânia - KEMPINAS, W. G. 2. GO, Brasil; ³Escola de Ciências Médicas, Farmacêuticas e Biomédicas, Pontifícia Universidade 1 Universidade Estadual de Campinas, Campinas - SP, Católica de Goiás, Goiânia - Goiás – Brasil. Brasil; 2 Universidade Estadual Paulista, Botucatu - SP, Brasil. Introduction: The pharmacokinetics properties, metabolism and toxicity are relevant parameters in the Introduction: Obesity is a serious health problem that search for new compounds with pharmacological has become a global epidemy. Among the indicated properties, since they are important features treatments, pharmacotherapy stands out. Several drugs responsible for most drug candidate failures. The have been approved in the last decades for this natural lignan hibalactone have been described with a purpose, however, some of them presented serious large variety of biological activities, such as risks, including for the reproductive system, and were antitumoral, antimicrobian and antioxidant, however withdrawn from the market. Locarserin (Belviq®) is a there is a lack of studies regarding its metabolism, as highly selective serotonin (5-HT) 2C receptor agonist well as its pharmacokinetics and toxicophoric drug that has been released recently for the long-term properties. Objective: To investigate the treatment of obesity with the promise of effectiveness pharmacokinetics and toxicity properties of the and low damage. The literature does not present any hibalactone metabolites using computational study of the impact of the drug on the reproductive approaches. Materials and Methods: MetaPrint2D- system. Objective: To investigate the impact of the React online tool was used to predict the metabolic treatment with locarserin on male rat reproductive pathways of hibalactone with simulations in phase I system. Materials and Methods: For a pilot study, adult and II of potential human biotransformations. The male Wistar rats were allocated (n=6/group) in control analysis of pharmacokinetics properties was performed group and treated with 0.5, 1 and 10 mg/Kg of with Molsoft DrugLikeness tool and its toxicological locarserin, in dark or light cycle. After 14 days of ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 126 treatment, the animals were evaluated for body and hours post-fertilization (hpf) using an inverted organ weights, food intake and sperm morphology and microscope. Lethality, sub-lethality and teratogenicity motility. Statistics: ANOVA or Kruskall Wallis indicators were used to assess the toxicity of CBD. The followed by Tuckey or Dunn’s test, p<0.05. Results: test was performed in triplicate. The results were The treatment with locarserin in the dark cycle did not statistically analyzed by ANOVA and Dunnett's test (α cause systemic toxicity as there was no change in body = 0.05). RESULTS: At tested conditions, no weight. In addition, the drug did not affect the malformations were observed at tested conditions until reproductive system in both kinds of exposure, since 144 hpf. CONCLUSION: This result suggests that the reproductive organ weights and sperm motility and CBD hasn’t teratogenic potential in the tested morphology were not affected. On the other hand, the concentrations. The tested concentrations were chosen treatment with the highest dose in the light cycle by weight proportionality between zebrafish embryos decreased the weight gain. Conclusions: These and human fetuses, and the lowest concentration tested preliminary results suggest that locarserin is effective is around 100 times higher than the proportional to lose weight, while do no damage to the male rat concentration. Larger range of concentration will be reproductive system. studied in the future steps of this study. Financial support / Acknowledgments: CAPES for Financial support / Acknowledgments: CAPES the financial support

EX 63 - CANNABIDIOL DOES NOT INDUCE EX 64 - DIISOPENTYL PHTHALATE (DIPEP) IS TOXIC EFFECT ON THE DEVELOPMENT OF A POTENT INHIBITOR OF FETAL ZEBRAFISH TESTICULAR TESTOSTERONE PRODUCTION IN RATS BRIGANTE, T.A. V.1.; ZUARDI, A.W.2; HALLAK, J.2; CRIPPA, J.A.2; OLIVEIRA, D.P.1. PASSONI, M. T. 1; CURI, T. Z. 1; ARAÚJO, G. 2; PALÚ, G. 2; CASTILHOS, B. S. 2; ZENDIM, L. A. 2; 1 School of Pharmaceutical Sciences of Ribeirão Preto MEYER, K. B. 2; VENTURELLI, A. C. 2; – University of São Paulo, Ribeirão Preto, SP - Brazil; DALSENTER, P. R. 1; MORAIS, R. N. 2; 2 Ribeirão Preto Medical School – University of São MARTINO-ANDRADE, A. J. 1,2. Paulo, Ribeirão Preto, SP – Brazil. 1. Departmento de Farmacologia, Laboratório de INTRODUCTION: Cannabidiol is a non-psychoactive Toxicologia Reprodutiva, Universidade Federal do compound of the Cannabis sativa that is being used for Paraná (UFPR), Curitiba - PR, Brasil; 2 Departamento the treatment of severe epilepsy and also studied for de Fisiologia, Laboratório de Fisiologia Endócrina e the treatment of several diseases of nervous system. Reprodutiva Animal, Universidade Federal do Paraná Since 2014, the prescription and importation of CBD is (UFPR), Curitiba - PR, Brasil. allowed in Brazil, and in January of 2017, ANVISA approved the registration of the first medicine based on Phthalates are high volume industrial chemicals found CBD. Toxicity studies of CBD have great relevance to in a variety of consumer products, including polyvinyl guarantee the safety of the population. In this context, chloride plastics, cosmetics, personal care products, we can highlight assays using zebrafish embryos food packaging, paints, toys, solvents, and considering that this organism has been widely used in formulations of pesticides and medicines. Many several toxicity studies, mainly teratogenicity and phthalates are known to be toxic to male reproductive neurotoxic studies due to its anatomical and genetic system, especially during critical prenatal windows of similarities with other vertebrates, including humans. androgen-dependent sexual differentiation. In a pilot OBJECTIVE: In the present work we evaluate the pregnancy cohort study conducted by our laboratory in potential of CDB to induce malformations on zebrafish Curitiba, we observed high exposure of Brazilian larvae exposed to doses similar to that expected for pregnant women to several phthalates, and in particular humans. MATERIAL AND METHODS: The Early to diisopentyl phthalate (DiPeP), which is in total Life Stage (ELS) Toxicity Test with zebrafish, based contrast to any other study population worldwide. The on OECD No. 236, was used to test the potential occurrence of DiPeP is a unique feature of Brazil, as it toxicity of CBD at concentrations of 5, 20, 70, 150 and is synthesized from isoamyl alcohol, a side product of 300 ng/mL. The fertilized eggs collected in the blastula sugarcane alcohol production. However, there is stage were individually arranged in 24-well plates. limited published data on the reproductive and Cannabidiol solutions, negative control and positive endocrine toxicity of DiPeP. In the present study, we control were prepared in standard water ISO-7346/3 investigated the effects of DiPeP on the ex vivo fetal with 0.01% of DMSO. Embryo and larvae testicular testosterone production in rats. Pregnant development were evaluated every 24 hours until 144 Wistar rats were exposed to vehicle (canola oil) and ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 127 three doses (125, 250 and 500 mg/kg/day) of DiPeP or testosterone. After five days of treatment, uteri were di-n-butyl phthalate (DnBP; reference) between days removed, dissected and weighted full and empty. 14 and 18 gestation, the masculinization programming Results: The results found in this study showed that the window in rats. At gestational day 18, fetal testes were vehicle group had no increase in absolute weight when collected and incubated in 0.5 mL of M-199 medium compared to the group treated with exemestane. for three hours at 37ºC and testosterone concentrations However, the group treated only with testosterone and measured by enzyme immunoassay in the medium. flutamide had a significant increase in uterine weight DiPeP and DnBP reduced testosterone production in a when compared to the vehicle group. After the dose-responsive manner, with significant reductions at statistical analysis of the groups treated with all three dose levels tested for DiPeP and at 250 and artesunate, none of them showed differences between 500 mg/kg/day for DnBP. The effective doses that the estrogenic control group. The analysis showed that reduced testosterone production to 50% of controls groups used as control had the expected results and (95% confidence interval) for DiPeP and DnBP were could be compared to the artesunate groups. The 93.1 mg/kg/day (62.9-139.3 mg/kg/day) and 225.5 estrogenic control presented an increase in uterine mg/kg/day (179.2-283.8 mg/kg/day), respectively. No weight with no statistical differences between groups signs of maternal or fetal toxicity were observed at any treated with artesunate, suggesting that artesunate does treatment group. Our results indicate that DiPeP, a not affect the enzyme. Conclusion: The results suggest phthalate that is relevant to the exposure scenario in that artesunate has no interference on aromatase Brazil, is a potent antiandrogen. enzyme activity.

Financial support / Acknowledgments: CAPES; Financial support / Acknowledgments: CAPES; CNPQ CNPQ

EX 65 - THE EFFECTS OF ARTESUNATE EX 66 - TOXICO-PHARMACOLOGIAL ACTIVITY OVER THE AROMATASE ENZYME EVALUATIONS OF THE SMALL-MOLECULE LQFM166: APOPTOSIS INDUCTOR AND GRECHI, N. 1; DA SILVA, G. N. 1; BOARETO, A. MDM2 ANTAGONIST C. 1; PASSONI, M. T. 1; ANDREOTTI, B. 1; CURI, T. Z. 1; MENEGATI, S. E. L. T. 1; DALSENTER, P. SANTOS, T. R. M.; SILVA, A. C. G 1; CARVALHO, R. 1. F. S. 2; MENEGATTI, R. 2; VALADARES, M. C. 1.

1 Universidade Federal do Paraná (UFPR), Curitiba - 1 Universidade Federal de Goiás; 2 Universidade PR, Brasil. Federal de Goiás.

Introduction: Artesunate is a medicine derived from Introduction: The antagonist of MDM2, the main artemisinin used for the treatment of malaria endogenous p53 inhibitor, by antitumoral compounds falciparum. It is used as therapy in pregnant women on has been emerged as a promising therapeutic target for the second and third trimester of pregnancy but not on the development of more effective agents. Objective: the first trimester due to teratogenic effects on the In this study, we designed, synthetized and embryo. Aromatase is an enzyme responsible for characterized the compound LQFM166 and convertion of androgen to estrogen. Objectives: This investigated the mechanisms of cytotoxicity and the study aimed to perform a screening of a possible acute oral toxicity. Methodology: Cytotoxicity profile artesunate activity over the aromatase enzyme. of LQFM166 was determined in K-562 cells, p53-null, Material and Methods: In this study, we examined the after 24, 48 and 72 hours of treatment using MTT in vivo effects of artesunate over the aromatase reduction assay. For mechanistic evaluation, leukemia enzyme using an adaption of the uterotrophic assay. cells were treated with 25% inhibitory concentration Immature rats were treated with three different doses (IC25) for 24 and 48 hours and assessed through flow of artesunate (6.25, 15 and 45 mg/Kg) associated with cytometry. The investigation of complexation of testosterone (3.5 mg/Kg) and flutamide (5 mg/Kg). p53/MDM2 was carried out in vitro by ELISA assay The groups used as controls were treated with: distilled using 3T3 cell lysates after treatment of 24 hours with water, testosterone plus flutamide plus exemestane (36 the compound. According the GD 129 the cytotoxicity mg/Kg) and testosterone plus flutamide (estrogenic in basal fibroblasts 3T3 was determined to estimate control). The group treated with exemestane was used DL50. Results: Outcomes showed that LQFM166 was as control due to its already known inhibition of cytotoxic for leukemia cells in a concentration-time- aromatase enzyme. Also, flutamide was administered dependent manner. Cell death mechanisms of to block testosterone receptors and to ensure that the LQFM166 in K-562 cells, revealed that the compound increased weight of the uteri was not related to induced cell cycle arrest and G1/G0 and G2/M phases, ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 128 promoting a significate up-regulation in p21, p27, chemically treated sham-operated controls presented cytochrome-c and Bax, while Bcl-2 and cyclin-B1 histological scores that varied between 1.02-1.03. were down-regulated. The p53-independent activity of Experimental groups presented significantly decreased the compound with an apoptotic profile was confirmed testicular weights and increased histological scores: by phosphatidylserine externalization and Caspase 3, AA/CPT-R groups - the score was 2.55 in the 16th and 7, 8 and 9 activation. In vitro studies showed that this 31st weeks, progressing to 3.48 at the 58th week; compound inhibits the p53/MDM2 interaction after DBP/CPT-R groups - 2.54 at the 16th week, 3.54 at the treatment. Furthermore, LQFM166 affect 3T3 cells 31st week and 3.63 at the 58th week. In control groups viability with the IC50 of 185,3 µM and estimated c-kit immunolabeling occurred only in basal LD50 of 706,7 mg/Kg (category 4 of GHS). spermatogonias. The experimental groups AA/CPT-R Conclusion: The rationally designed prototype and DBP/CPT-R presented increased c-Kit LQFM166 induced cell death in leukemic cells by immunoreactivity in all moments of study but this was activation of several pathways related to MDM2 significant only for AA/CPT-R animals at the 31st antagonism, independently of p53 function. (12.5%) and 58th (32.1%) weeks. Conclusion: The score system adopted for histological evaluation Financial support / Acknowledgments: CNPq allowed assessment of testicular damage by chemicals in cryptorchid rats. The c-Kit protein is a surface membrane tyrosine kinase receptor required in various EX 67 - TESTICULAR HISTOLOGICAL stages of normal spermatogenesis. In the present model ALTERATIONS AND c-Kit the spermatogenesis process was severely disturbed; IMMUNOEXPRESSION IN CRYPTORCHID the increased number of tubules with c-Kit positive RATS EXPOSED IN UTERO AND germ cells was predominant in AA/CPT-R animals. POSTNATALLY TO ACRYLAMIDE OR DI-N- BUTYL-PHTHALATE Financial support / Acknowledgments: FAPESP (Process No. 2012/09873-4), CAPES and TOXICAM. LIMA, T. R. R.; MIOT, H.A.2; SOUZA, N.P.1; CARDOSO, A.P.F.1; GOMIDE, L.M.M.1; DE CAMARGO, J.L.V.1; NASCIMENTO, M.G.1. EX 68 - CREATINE KINASE ACTIVITY AND NPSH LEVELS ARE MODIFIED IN C6 CELLS 1 Universidade Estadual Paulista (UNESP), Faculdade EXPOSED TO GLYPHOSATE-BASED de Medicina, Departamento de Patologia, Núcleo de HERBICIDE Avaliação do Impacto Ambiental sobre a Saúde Humana (TOXICAM), Botucatu - SP, Brasil; UEDA, C.N.1; CAPPELLARO, L.G.1; RODRIGUES, 2Universidade Estadual Paulista (UNESP), Faculdade L.1; REMOR, A.P.2; LATINI, A.3; GLASER, V.1. de Medicina, Departamento de Dermatologia e Radioterapia, Botucatu - SP, Brasil. 1 Coordenadoria Especial de Ciências Biológicas e Agronômicas, Centro de Ciências Rurais, Introduction and objective: Cryptorchidism and in Universidade Federal de Santa Catarina, Curitibanos- utero exposure to xenobiotics are risk factors for male SC. 2 Programa de Pós-Graduação em Biociências e reproductive tract disorders. This experimental study Saúde, Universidade do Oeste de Santa Catarina, aimed to evaluate the testes of rats exposed in Joaçaba-SC. 3 Departamento de Bioquímica, Centro de utero/postnatally to acrylamide (AA) or to di-n-butyl- Ciências Biológicas, Universidade Federal de Santa phthalate (DBP) and also submitted to Catarina, Florianópolis-SC. cryptorchidism/orchiopexy. Material and Methods: Dams were exposed by gavage to AA 10 mg/kg/day or Introduction: Glyphosate and paraquat are part of DBP 500 mg/kg/day during gestation and by diet many commercial herbicides. Some studies have during lactation. At the 21st postnatal day, male pups demonstrated that these agrochemicals present were submitted to surgical cryptorchidism (CPT) and cytotoxicity, such as the exacerbated production of after 3 weeks to orchiopexy (CPT-R), but continued reactive oxygen species (ROS). ROS can interact with under exposure to xenobiotics by diet until they were proteins and change their activity and, in this context, 16, 31 or 58 week-old. The testes were weighed and it is known that creatine kinase (CK), an enzyme submitted to a histological score from minimal (Class involved in energy metabolism, is highly susceptible to 1) to severe alterations (Class 4). c-Kit ROS because it has a thiol group in its active site. immunoexpression was qualitative and quantitatively Objectives: The aim of the present work was to assessed by the location of positive germ cells within analyze CK activity, non-protein thiol group (NPSH) the seminiferous epithelium and the number of positive levels and ROS generation in astroglioma cells tubules/total seminiferous tubules. The level of exposed to glyphosate or paraquat based herbicides. significance was considered as p<0.05. Results: Non- Material and methods: Rat astroglioma cells (C6 cells) ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 129 were exposed for 24 hours to glyphosate-based or coagulation, eye and body pigmentation, somite paraquat-based herbicides (0-80 μM). After the formation, tail circulation, hatching, among others. incubation period, CK activity and NPSH levels were Subsequent to the hatching there were observed and measured. In addition, intracellular ROS generation reported, mortality, edema, posture, spine deformities was analyzed using 2,7’-dichlorofluorescein-diacetate and undersized embryos. In mice, the substances did (DCF-DA) and superoxide generation was measured not cause mortality and did not cause changes in using dihydroethidium (DHE). Results: CK activity behavior or signs of toxicity, such as diarrhea, ptosis, has decreased in cells exposed to 80 μM of glyphosate- piloerection, alertness. Macroscopic analysis of organs based herbicide, but not in paraquat-based herbicide. of Swiss mice showed no changes in their color, ROS generation was not modified by the treatments. morphology and no hemorrhagic points. To confirm However, NPSH levels have increased in C6 cells these results, histological analysis will be realized. The exposed to glyphosate-based herbicide. Conclusion: next step will contemplate the dose of 2000 CK can be a glyphosate-based herbicides target and the mg/kg/animal. In zebrafish embryos, even high increasing of NPSH levels can be a protective cellular concentrations of both ruthenium complexes did not mechanism to avoid ROS production and oxidative cause toxicity in their development and did not cause stress. mortality. These data suggest that these substances have low toxicity, and may be promising prototypes of Financial support / Acknowledgments: CNPq drugs against breast cancer. Financial support / Acknowledgments: CAPES

EX 69 - SECURITY ANALYSIS OF TWO NEW CANDIDATES OF CHEMOTHERAPEUTIC EX 70 - HISTOMORPHOMETRIC ANALYSIS BASED ON RUTHENIUM COMPLEXES FOR OF THE CORNEAS IS AN ADDITIONAL THE BREAST CANCER TREATMENT PARAMETER TO REFINEMENT OF BCOP TEST VELOZO-SÁ, V. S.; MELLO-ANDRADE, F. 1; LIMA, A.P. 1; GOVEIA, R.M. 1; PEREIRA, L.R. 2; ANDRADE W.M.; ANDRADE, WA1; SILVA, BATISTA, A.A. 2; SILVEIRA-LACERDA, E.P. 1. ACG1; VALADARES, MC1.

1 - Universidade Federal de Goiás, Goiânia-GO, 1 Universidade Federal de Goiás, Goiânia – GO, Brasil; 2 - Universidade Federal de São Carlos, São Brasil. Paulo, Brasil. Introduction: The Bovine Corneal Opacity and LGMC-68 and LGMC-69 are ruthenium complexes Permeability (BCOP) assay is an alternative method that have been active against breast cancer cells in widely used to assess the ocular irritation/corrosion vitro. It was proposed in this study to evaluate toxicity potential of chemicals and mixtures. The standard and safe of these substances, through the acute oral BCOP test provides information about permeability toxicity in Swiss mice (OECD 423) and and opacity, however, histomorphometric analysis of embryotoxicity in zebrafish model (OECD 236). In the the cornea have been recommended as an additional acute oral toxicity, fasting animals were weighed and parameter to better categorize eye irritant products, in the test substances administered orally in a single special to differentiate eye irritants of Category 1 from initial dose (300 mg/kg/animal) by gavage. These Category 2A and 2B. Though such analysis associating animals were observed individually every 30 min the depth of corneal damage induced by test substances during first 4h and every two days until the 14th day. and severity of damage, was not scientifically The parameters observed were: death, sedation, demonstrated as an additional parameter for refinement alertness, diarrhea, convulsions, spontaneous motor of the BCOP test. Objective: The aim of this study was activity, among others. On day 14th, the animals were to measure the depth of injury to the bovine cornea euthanized and the liver, kidneys, spleen, lungs and induced by eye irritant substances from all Globally heart were removed, washed with PBS, their relative Harmonized System of Classification and Labelling of weights were determined, and fixed in 10% (v/v) Chemicals (UN GHS) categories and associate it with buffered formalin. In zebrafish test, eggs were severity of the damage. Material and Methods: For the collected within 30 min after natural mating. Test proficiency, 12 substances belonging to UN GHS started with newly fertilized eggs, exposed to categories (Category 1, 2A and 2B) were selected. concentrations 0.5, 1.2, 2.9, 7.1, 17.1, 41.4 and 100 mg BCOP assay was performed according to OECD Guide L-1 of LGMC-68 and LGMC-69, and vehicle (DMSO, 437 of 2013 for in vitro irritancy score (IVIS) 0.1%). Twenty eggs per treatment were distributed in determination. Additionally, after the assay of 24-wells microplates in triplicate. In the embryo phase, permeability with fluorescein, the corneas used were the following parameters were evaluated: egg included in Tissue-Tek O.C.T compound at -20 ° C, ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 130 sections of 5 μm thickness were obtained in the cryostat, which were stained with the nuclear dye Hoechst 33342 at 0.5 μm/ml. Each section was analyzed by fluorescence microscopy and 5 measurements of permeation depth of fluorescein were obtained relative to the total thickness of the epithelium. Results: The results showed that the fluorescein permeation depth was directly proportional to the degree of irritation of the substances tested. Severe irritants, Cat 1, promoted the highest rates of permeation followed by moderate and mild irritants. Non-irritant substances, in turn, showed similar permeation indexes to negative control.Conclusion: The refinement by histomorphometric analysis of the epithelial permeation of fluorescein could be considered a useful quantitative endpoint to better categorize eye irritants substances.

Financial support / Acknowledgments: CNPQ CAPES / FACULTY OF PHARMACY - UFG

FOOD TOXICOLOGY ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 132

FO 01 - COMPARATIVE MICROBIOLOGICAL microbiological quality control before the arrival to ANALYSIS BETWEEN MINERAL AND consumption of students and workers. DRINKING WATER FOUNTAIN CONSUMED Financial support / Acknowledgments: Laboratório AT THE FEDERAL UNIVERSITY OF GOIÁS de Controle de Qualidade Microbiológico de Medicamentos, Faculdade de Farmácia - Universidade VINHAL, D.C. 1,2; PEREIRA, C.S.P. 1; BRASIL, Federal De Goiás (UFG); Faculdade De Inhumas A.M.P. 2; MOURA, D.B.C. 1. (FACMAIS).

1. Laboratório de Controle de Qualidade Microbiológico de Medicamentos, Faculdade de FO 02 - EPIDEMIOLOGICAL OUTLINE OF Farmácia – Universidade Federal de Goiás (UFG) – EXOGENOUS INTOXICATION ASSOCIATED Av. Universitária, St. Leste Universitário, CEP 74605- WITH AGROTOXICS IN GOIÁS, BETWEEN 170. 2. Faculdade de Inhumas (FACMAIS) – R. 2011 AND 2016 Sebastião Mota, n. 253, Inhumas – GO, CEP 75400- 000. NASCIMENTO, H. A. 1; MAYA, A. C. M. 1; MORAIS, L. T. DE 1; SILVA, L. A. 1; MACHADO, Introduction Water often use in Brazil for human H. C. P. 1; PRADOS, V. O. 1; SILVA, L. O. 1; consumption is generally obtained from different MARTINS, I. L. 1; LIMA, G. C. DE 1; OLIVEIRA, R. underground sources. The good visual appearance of R. 1. this gives consumers a sense of purity, but potability is not always perceptible to vision or smell, and a 1. Unievangélica laboratory analysis is necessary to detect possible microbial contaminations. Objective This study has the Introduction: Brazil is a featured country in the world objective to evaluate and compare the microbiological as the biggest consumers of agrotoxics, responsible for quality of water by monitoring a commercial bottle of 86% of total use in Latin America. Intoxication caused mineral water and a public water source located and by agrotoxics come from a complex interaction consumed at the Federal University of Goiás, Goiânia. between the product attributes and the farmer Material and Methods Twenty-two samples were exposure, or lately, ordinary consumer. The most collected from the commercial bottle of mineral water important absorption way is by the skin, and vast and the public water fountain, in the period between majority of cases result in emergency treatment, April and May 2017 every other day. Were analyzed especially in the public health system. Objective: The according to what is recommended in the concierge n. objective was trace a risk and exposure outline of 2.914 of December 12, 2011. Plate count agar was people affected by agrotoxics in the state of Goiás, used for the counting of total bacteria and specific agar between 2011 and 2016. Materials and methods: This for the research of pathogenic microorganisms is a retrospective, qualitative and quantitative paper, represented by Pseudomonas aeruginosa, Escherichia based on active search of data in multiple databases coli, total coliforms and thermotolerant. Results The and a literary review about the subject. Were used analyzed waters passed through an on-going process of major criteria for mortality and intoxication – with reading and strains, so for all mineral water sample registered cause by agrotoxic – since 1996, up to build was obtained score higher than 500 UFC/ml and the a demographic evolution in both state and country. microorganism found was gram negative rods and Results: It was revealed that Goiás is responsible for indole negative not characteristic of E. coli. Specific almost 52% of total cases, occurring especially in research was done for all samples of mineral water urban zones, by different reasons, being the principal through enrichment in tryptone soya broth followed by suicide attempt and labor exposure. The most affected the strain in agar cetrimide. So, it was obtained age is between 20 and 39 y.o., representing 25% of greenish colonies, presenting gram negative rods and cases, mostly in males.Conclusion: Even dealing with positive oxidase, which was confirmed through the the lack of registers, it was possible to delimitate a biochemical test (BD BBL) the presence of P. map that exposes the number and quality of cases aeruginosa in mineral water. However, the legislation occurred in each state sector. recommended results below of 500 UCF/mL and Financial support / Acknowledgments: absence in 100 mL. Conclusion From the results Unievangélica found, we concluded that the mineral water consumed in the UFG showed microorganism counts higher than those recommended by the legislation and pathogenic microorganisms characterized by the presence of P. aeruginosa indicating the low microbiological quality of the water samples, showing the need of a strict

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 133

FO 03 - EPIDEMIOLOGICAL OUTLINE OF FO 04 - NITRATE IN LETTUCE PURCHASED EXOGENOUS INTOXICATION ASSOCIATED AT SUPERMARKETS IN THE CENTRAL WITH FOOD AND DRINKS IN GOIÁS, SEMIARID REGION OF CEARÁ BETWEEN 2011 AND 2016 ARAUJO, D.D.C. 1; CUNHA, L.S. 1; SILVA, E. 1; NASCIMENTO, H. A. 1; MAYA, A. C. M. 1; FILHO, D.M.B. 1; FREITAS, Y.P.F 1; MORAIS. MORAIS, L. T. DE 1; SILVA, L. A. 1; MACHADO, I.C.O. 1. H. C. P. 1; PRADOS, V. O. 1; SILVA, L. O. 1; MARTINS, I. L. 1; LIMA, G. C. DE 1; OLIVEIRA, R. 1. Centro Universitário Católica de Quixada- R. 1. UNICATOLICA, Quixada-CE, Brasil.

1. Unievangélica Introduction Vegetables constitute a vital part of the human diet, being the main source of minerals, Introduction: Food poisoning or intoxication is defined vitamins, dietary fiber and phytochemicals. Green as a pathology caused by direct consume of toxic leafy vegetables are foods that contain considerable agents in food or water contaminated by bacteria, amounts of nitrate, which can have both positive and fungus, virus and other microorganisms or ingestion of negative effects on the human body. Nitrate levels in their respective toxin, as metals and chemicals present vegetables depend on the type of plant, temperature, directly in food. Additionally, infection associated with and moisture and nitrogen fertilization. Depending on intoxication are serious grievances caused by the amount ingested, nitrates can promote the infectious agents, like parasites an some bacteria able methaehemoglobinaemia in children, an illness that to secrete their toxins after inoculation in the human decreases the capacity of the blood to transport organism or spending part of their life cycle in it. oxygen. Nitrite can react with the amines present in the Objective: The objective was to reproduce an food forming nitrosamines, one of the most potent epidemiological outline of food poisoning cases in carcinogenic discovered until now. The objective is to Goiás. Materials and Methods: It’s a descriptive and determine and to compare the nitrate content in lettuce retrospective paper, based on active search of acquired at different supermarkets in the central epidemiologic data of intoxications in the state, from semiarid region of Ceará, from organic and 2011 to 2016, in DATASUS and Sinitox databases, conventional farming, using the method of Cataldo et apart literary review about the subject. Results: al. (salicylic acid). The measurements were done Between 2011 and 2016 were registered 1128 cases of treating the plant extract with active carbon and using food poisoning in Goiás, whit 13% of all cases in the the untreated samples for comparison purposes. A country coming from Goiás, with an incidence of calibration curve was linear (20-300 μg). Results: The around 220 cases per year. The lethality observed is samples with the highest concentrations of low and predominant in children and old age. Most of conventional lettuce presented concentrations of nitrate cases and exposition occurred on men in economically ranging between 16.190,3 and 14.381,2 ppm. Organic active age (20 to 49 y.o.). Predisposing factors for lettuce presented concentrations between 1.923,6 and major exposure and harm were minor scholarship (up 1.488,5 ppm. The levels of nitrates were higher in to 7 years resulting in 50% of registers), urban conventional farming compared to the organic one. residence and old age. Conclusions: The obtained Conclusion: The results indicate that the analyzed results allowed delimitating a risk outline to state lettuce from conventional farming contains significant population, calculate the exposure in each sector, amounts of nitrate in their composition, but it is still an chances of mortality and delimitate a representative acceptable intake into the human body. The daily adult map of the number of cases for each state sector. consumption of 100 g amounts of the studied lettuce Financial support / Acknowledgments: was found not exceeding the Acceptable Daily Intake Unievangélica for nitrates. Financial support / Acknowledgments: Centro Universitário Católica de Quixada- UNICATOLICA

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FO 05 - METHODOLOGY FOR THE FO 06 - TOXICOLOGICAL TESTING ESTIMATION OF PESTICIDE RESIDUES IN RATIONALE FOR ACID AND/OR ESTER ANIMAL COMMODITIES FOR THE FORMS OF AN AGROCHEMICAL: ARYLEX™ BRAZILIAN SCENARIO - ESTABLISHMENT AS A CASE STUDY OF MAXIMUM RESIDUE LIMITS AND CONDUCTION OF DIETARY RISK MANOJ AGGARWAL1, CLAIRE TERRY2, ASSESSMENTS JYOTIGNA MEHTA1, REZA RASOULPOUR2 AND DALAND JUBERG2. VALERIO, L. B.; BRISIGHELO, L.V 1; NASCIMENTO, E. S. 2. 1. Dow AgroSciences Ltd, Oxfordshire, UK; 2. Dow AgroSciences LLC, Indianapolis, USA. 1. Universidade de São Paulo, São Paulo – SP, Brasil; 2. Universidade de São Paulo, São Paulo. There are many agrochemicals where the active substance exists in multiple forms of esters and/or salts INTRODUCTION: In Brazil the Maximum Residue such as 2,4-D, triclopyr, fluroxypyr, aminopyralid and Limit (MRL) for pesticides is established by the others. In such situations, a scientific rationale for competent authorities only for plant commodities and toxicological testing which follows the principles of it is published in the ANVISA’s active ingredients 3Rs (Replacement, Reduction and Refinement) in monographs. MRLs on commodities of animal origin animal research has been applied. A common scenario are not established, and therefore are not considered in is that the ester/salt form of a chemical is hydrolysed in the calculation of exposure. Consequently, the actual mammals into the acid form so that systemic (post- values of pesticide residues in animal commodities are hepatic) exposure is to the acid i.e., ester/salt form is not known, becoming a limitation for the conduction bio-equivalent to acid form. In such cases, the acid of a more accurate exposure assessment of the form can be used as the main test material for the Brazilian population to pesticides. The MRLs entire toxicological package, while the ester and/or salt established by the JMPR (Joint Meetings on Pesticide can be tested in select studies to determine Residues) and OECD (Organization for Economic Co- toxicological equivalence (e.g., genotoxicity and sub- operation and Development) for animal commodities acute study). As a result, each ester or salt does not cannot be used in the first instance as being need animal-intensive like carcinogenicity or representative for Brazil, since a thorough analysis is reproductive toxicity studies. Reference Doses (RfDs) needed in order to identify if they reflect the Brazilian can be determined using the acid studies, which are scenario. OBJECTIVE: Propose a methodology to be applicable to ester and salt for human health risk used for the Brazilian scenario to estimate pesticide assessments. Halauxifen-methyl (Arylex™) is a new residues levels in animal commodities,creating Excel® herbicide which is rapidly and completely metabolised calculators to aid in estimating the animal dietary into halauxifen acid in mammals. Halauxifen acid only burden for each animal and estimating the residues in was found in blood of rats orally given halauxifen- each animal commodity. METHODS: i) Obtaining methyl (i.e., acid and ester forms were bio-equivalent). information about Brazilian animal feeding practices: Therefore, halauxifen acid was tested for the entire through research in national references and battery of toxicological studies as required by global consultation with experts. The information needed to regulations. Halauxifen acid mainly caused kidney create the animal feed table will follow the OECD toxicity at relatively high doses. When halauxifen- Guidance Document on Residues in Livestock (2013); methyl was tested for acute-, geno- and ii) Construction of the calculators to estimate the developmental-toxicity studies, it was found to be animal dietary burden and the pesticide residue level in equivalent to halauxifen acid. However, in a sub-acute animal commodities: will be done through Excel® and toxicity study halauxifen-methyl caused liver toxicity, will follow the instructions in the same OECD i.e., toxicologically not equivalent for target organ guidance (2013), OECD guideline 505 and FAO toxicity. For these types of chemicals, comprehensive Manual (2009); iii) Validation of the calculators will Mode of Action (MoA) studies were performed to be done using the examples contained in the OECD elucidate liver toxicity caused by halauxifen-methyl. It guidance (2013) and FAO Manual (2009). RESULTS was found that halauxifen-methyl at high doses AND DISCUSSION: The results obtained so far were: induces nuclear receptor activation in the liver but the construction of the Brazilian animal feed tables, halauxifen acid does not. A weight-of-evidence derived from crops, for different types of animals; the approach was taken to show that MoA-based RfDs for identification of the predominant production system in halauxifen-methyl were acceptable for human health the country - which is the extensive system; and the risk as. construction of the animal dietary burden calculator. Financial support / Acknowledgments: Dow CONCLUSION: work in progress. Agrosciences Financial support / Acknowledgments: Syngenta ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 135

FO 07 - MODE OF ACTION AND IPCS HUMAN Financial support / Acknowledgments: Dow RELEVANCE FRAMEWORK FOR Agrosciences. HALAUXIFEN-METHYL (ARYLEX™) INDUCED RODENT LIVER EFFECTS FO 08 - DATABASE CREATION OF RAW FOOD CONSUMPTION OF THE BRAZILIAN MANOJ AGGARWAL1, CLAIRE TERRY2, LYNEA POPULATION THROUGH THE RESEARCH OF MURPHY3, MATTHEW LEBARON3 JYOTIGNA FAMILY BUDGETS MEHTA1, REZA RASOULPOUR2 AND DALAND JUBERG2. PAIS, M.C.N 1; BRISIGHELO, L.V 2.

1. Dow AgroSciences Ltd, UK; 2. Dow AgroSciences 1. Ferst Consulting; 2. University of São Paulo. LLC, Indianapolis; 3. The Dow Chemical Company, Michigan. INTRODUCTION: The Brazilian Institute of Geography and Statistics (IBGE) is the body Halauxifen-methyl (Arylex™), a novel herbicide, responsible for the development of Family Budget induced rodent liver enlargement, hypertrophy, and Surveys (POF), which allows evaluating, among other hyperplasia via an Nuclear Receptor (NR) mediated data, the trend of food consumption of the Brazilian Mode of Action (MoA) with the following WHO population. However, the way in which th data was International Programme on Chemical Safety Human published by the IBGE does not allow its immediate Relevance Framework (IPCS-HRF) key events: #1) use to carry out human risk assessments through diet, pre-systemic liver exposure to halauxifen-methyl, #2) since the foods were shown in their final form of NR activation, following binding of halauxifen-methyl consumption. OBJECTIVE: Creating a database by with the receptor, with subsequent liver weight compiling data from the POF-2008/2009, to obtain the increase and hepatocyte hypertrophy, and #3) individual average daily consumption of raw hepatocellular proliferation, i.e., hyperplasia. From the commodities of the Brazilian population to be used in 90-day integrated MoA and toxicokinetic rat study it the dietary risk assessment process. METHODS: 1) was found that, for key event #1, low levels of Dismembering Recipes Spreadsheet: the food halauxifen-methyl were present in liver after dietary ingredients included in the consumed recipes by exposure; there was almost no post-hepatic (i.e., Brazilian population were classified and dismembered systemic) exposure to the halauxifen-ester as it was as follows: i. Raw food; ii. Prepared foods; iii. rapidly metabolized in liver to halauxifen acid, which Processed foods. 2) Food consumption Spreadsheet: was not an NR activator and was excreted in urine. For Personal food consumption data from POF were key event #2, NR activation, measured by Cyp1a1 collected from all residents (≥ 10 years old) for two induction, and correlated with liver hypertrophy nonconsecutive days, in order to obtain the average occurred at ≥ 50 mg/kg/day halauxifen-methyl. For daily food consumption per capita for the major key event #3, hepatocellular proliferation was regions of Brazil. 3) Correlating the spreadsheets and observed with exposure to 261 mg/kg/day halauxifen- Creating a Database: the average per capita food methyl. A threshold for NR activation and liver effects consumption spreadsheet for each of the major regions occurred at 10 mg/kg/day halauxifen-methyl (NOAEL was cross-checked with the recipes spreadsheet. from rat 90-day study). Collectively, the data provided Afterwards, a program was developed with the Delphi a high level of confidence that halauxifen-methyl 2010 platform. RESULTS: based on this approach, a induced rodent liver effects through an NR-mediated program was created: “Analysis of the average per MoA at doses >10 mg/kg/day. This NR pathway is capita food consumption of the Brazilian population”, conserved across species and prototypical NR ligands in which it is possible to obtain information regarding (i.e., metabolically stable, resistant to degradation, the daily consumption of prepared food and of each high binding affinity for the receptor) are activators of raw food that compose them. CONCLUSION: through both human and rodent NR; however, halauxifen- this tool, it is possible to establish real values of methyl is not a prototypical receptor ligand as it was average daily consumption per capita for all foods, rapidly metabolized (metabolically labile) to considering their consumption raw and the halauxifen acid (not a ligand for NR) and as compared contribution of this food in the composition of the to rodents, humans were found to be less sensitive to consumed recipes which is of great relevance for NR activations and associated effects. A weight-of- toxicological interest, when assessing the dietary risk evidence approach was taken to show that MoA-based assessment. ACKNOWLEDGMENTS: ILSI BR (GT- RfDs for halauxifen-methyl were acceptable for human POF); Carlos Ledo (Embrapa); Paulo Yatsuzuka. health risk assessments. Importantly, this approach has Financial support / Acknowledgments: ILSI Brasil been ratified by regulatory agencies from US, Canada, (GT-POF); Carlos Ledo (Embrapa); Paulo Yatsuzuka; EU, Australia and New Zealand. Ferst Consulting.

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FO 09 - ACUTE TOXICOLOGICAL TESTING FO 10 - CHEMICAL COMPOSITION OF OF BIOMASS OF RUBRIVIVAX CURCUMA LONGA L. ESSENTIAL OIL GELATINOUSUS EXTRACTED FROM DIFFERENT HARVESTS AND ITS ANTIFUNGAL ACTIVITY AGAINST PEREIRA, R. C.; PIRES, H.1; PONSANO, E.H.G1.; FUSARIUM GRAMINEARUM MINGATTO, F.E2.; ROZZA, D.B.1; PERRI, S. H. V.1. HOELTGEBAUM, D. 1; ROMOLI, J. C. Z. 1; BREDA, J. 1; NIEVIEROWSKI, T. H. 1; PANTE, G. 1. Universidade Estadual Paulista (Unesp), Faculdade C. 1; FERREIRA-ROMANICHEIN, F. M. D. 1; LINI, de Medicina Veterinária, Araçatuba; 2. Universidade R. S. 1; GLORIA, E. M. 2; MOSSINI, S. A. G. 1; Estadual Paulista (Unesp), Faculdade de Ciências MACHINSKI, JUNIOR M. 1. Agrárias e Tecnológicas, Dracena. 1. Department of Health Basic Sciences, State The slaughter of animals for food processing stands University of Maringá, Maringá, Paraná, Brazil; 2. out by the abundant use of water. Brazil has great Department of Agroindustrial Science and renown in the scenario of animal protein production, Technology, University of São Paulo, College of therefore, worrying amounts of effluents are produced. Agriculture Luiz de Queiroz, Piracicaba, São Paulo, Biotechnological processes, such as the use of Brazil. photosynthetic bacteria, are widely studied to help solve this problem in a sustainable way. The Introduction: Essential oils (EOs) have become a great degradation of the organic effluent load by the alternative for synthetic fungicides because they are microorganism Rubrivivax gelatinousus results in the natural, biodegradable and present low toxicity. Most production of a biomass of technological value, since it EOs extracted from aromatic plants have antifungal has carotenoids. With the future interest of using this and anti-mycotoxigenic activity. The variability in biomass as a food additive, the aim of this work was to chemical composition of EOs can include soil evaluate the in vivo toxicity of the biomass. Female variations, harvest year, climatic conditions, plant part Wistar rats weighing between 180 and 200 g were extracted and extraction techniques. Aims: Compare randomly divided into: control group (corn oil), B1 the chemical composition of Curcuma longa (turmeric) (300 mg biomass/kg body weight), B2 (1000 mg EOs (TEOs), harvest in 2015 and 2016 (TEO15 and biomass/kg body weight) and B3 (2000 mg biomass/ TEO16), and its antifungal activity against Fusarium kg body weight) and received single dosing via graminearum, a toxigenic strain. Methods: The gavage. Doses were determined according to the Acute chemical composition of TEO15 and TEO16 extracted Toxic Class Method for acute toxic dose testing by hydrodistillation, (purchased from Cooperativa de (Guideline 423). The statistical significance of the Açafrão de Mara Rosa, Goiás, Brazil), was identified parameters of the animals (weight, relative body by gas chromatography coupled with mass spectra. weights, feed intake, water consumption, excreta and The minimum inhibitory concentration (MIC) against temperature) was evaluated by ANOVA followed by F. graminearum was determined for both oils. Results: the Dunnet test for comparisons of the treated groups The main components of TEO15 and TEO16 were β- in relation to the control, and results with P < 0.05 turmerone 25,90% e 28,48%; α-turmerone 18,16% e were considered statistically significant. The results of 18,79% e ar-turmerone 23,75% e 21,94%, the Hippocratic screening were submitted to the respectively. The TEOs presented the same MIC Kruskal-Wallis test to compare the mean scores against F. graminearum at concentration of 1250 obtained. Significant difference (P < 0.05) was µg/mL. This result was probably due to the similar observed only in the increase of the production of percentages of the main components of TEO15 and urine of group B2. Observational analysis did not show TEO16. Conclusions: The TEOs from different harvest any behavioral changes (P > 0.05), that is, there was no showed the same main components in similar visible impairment of the respiratory and nervous percentages. The antifungal activity of TEOs was systems of the animals. The kidneys showed no proved against Fusarium graminearum possibly a microscopic changes, while degeneration and necrosis result of the synergistic action of its chemical of hepatocytes were observed microscopically in the components. animals in the higher dose group (B3). There was no Financial support / Acknowledgments: Coordenação death of any animal during the treatment, and it was de Aperfeiçoamento de Pessoal de Nível Superior not possible to establish the LD50 of the biomass. Our (CAPES). results indicate that biomass presents low toxicity since there were no recorded deaths and few alterations were observed in treated animals.

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FO 11 - BRAZILIAN CHRONIC RAW FOOD CONSUMPTION: A COMPARATIVE ANALYSIS FROM 1995 TO 2009

LUPIANHEZ, V.; PAIS, M. 1; VALÉRIO, L. 2; GUIMARÃES, S. 3.

1. Ferst Consultoria; 2. Syngenta; 3. BASF.

INTRODUCTION: The Family Budget Surveys (POF), carried out by the Brazilian Institute of Geography and Statistics (IBGE), collect household food acquisition and/or consumption data since the 70s. There are differences among the methodologies. The 1995-96 POF obtained a per capita household food consumption, by product’s average price. Surveys of 2002-03 and 2008-09 registered a daily food acquisition by residence. The last one published for the first time the individual consumption. In 2015, a Working Group from ILSI (International Life Science Institute) – Brazil developed a database of raw food consumption, from 2008-09 POF, dismembered in percentile of ingredients. OBJECTIVE: Conducting a comparative analysis of chronic consumption of raw commodities from the Brazilian population, through data from the last three POFs, and from ILSI’s program to assure the reliability of the available exposure data when assessing dietary risk. METHODS: i)Data analysis of the last three POFs to identify differences between the surveys methodologies; ii)Data consumption extraction from IBGE and ILSI’s program; iii)Clustering the crops according to Joint Normative Instruction (INC01) to compare the crop consumptions; iv)Comparing the two ILSI’s methodologies to IBGE publication identifying the critical scenario; v)performing the sum of crop group consumptions to compare Brazilian chronic food consumption data from 1995 to 2009. RESULTS AND DISCUSSION: The unrefined data from 2008-09 POF reflect lower food consumption and between the two ILSI’s methodologies there were no significant differences. The report of non-consecutive two-day consumption provides a more appropriate analysis. Among the crop groups, there is trend of consumption over the years, and food consumption of the Brazilian population has been increasing, with some exceptions. CONCLUSION: It was possible to identify a trend in food consumption of the Brazilian population. It was also possible to verify the accuracy of the refined data from ILSI program on chronic food consumption, which may guarantee the use of these data with greater confidence in dietary risk assessments, being of great toxicological relevance. Financial support / Acknowledgments: Ferst Consultoria

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FO 01 - ADVANTAGES OF THE ALTERNATIVE Financial support / Acknowledgments: BIOLOGICAL MATRIX IN FORENSIC Departamento de Patologia - UFSC; Instituto Geral de TOXICOLOGY ANALYZES: COCAINE Perícias de Joinville - IGP/SC. DETECTION IN BLOOD, URINE, AND HAIR FO 02 - ALCOHOL AND DRUGS IN SUICIDAL MAIA, A. H. A. M.; CRUZ, A. C. H. 1; ESPÍNDOLA, ETIOLOGY AUTOPSIES PERFORMED IN THE B. 1; OLIVEIRA, E. C. 1; PERIN, M. 1; PERICOLO, ATACAMA REGION - CHILE, 2007 TO 2016 S. 2. CRUZ, C.E.; ESPINOZA C.M. 1, ARANEDA E.C. 1, 1. Universidade Federal de Santa Catarina, SILVA C.A. 2, NOVAKOVIC I.M. 2, FLORES Florianópolis - SC, Brasil; 2. Instituto Geral de D.R.1. Perícias de Joinville, Joinville - SC, Brasil. 1. Laboratory of Forensic Medical Service of Copiapó, Introduction: The use of recreational cocaine (COC) is Chile. 2. Unit of Thanatology of Forensic Medical related to violence. Thus, it is an important analyte for Service of Copiapó, Chile. forensic toxicology. In this context, the most common samples are blood and urine. In bodies in advanced Alcohol and drugs are closely linked to suicides, and decomposition stage, sometimes they are not available this cause of death is a public health problem for analysis. In this case, alternative matrices, such as worldwide. The aim of the present study was to hair, are very useful. A slow decomposition compared describe the toxicology in autopsies of suicidal to other tissues is also an advantage. Objectives: To etiology, in the Atacama Region - Chile, between 2007 quantify COC in hair samples and compare these and 2016. This descriptive-retrospective study of the results with those of blood and urine. Materials and last 10 years yielded the following results: from 1637 methods: 22 cases of violent death from General autopsies studied, the 20,1% correspond to deaths of Institute of Skills of Santa Catarina (IGP/SC) were suicidal etiology (n=330), the average age was 39 evaluated. Hair samples were washed with a tween, years, 83% were men. Alcohol was detected dichloromethane, and water. Fragmentation was (≥0.20g/L) in 45.8% of autopsies due to suicide of performed followed by pulverization. For digestion death, with an average of 1.64 g/L ethanol, the average was used methanol and 0.1M hydrochloric acid at being in men 1.66g/L and in women 1.50g/L. Of the 45°C overnight. A Solid Phase Extraction was carried total positive cases for ethanol, 84% were in the out. The analysis was performed by High-Performance drunkenness state (≥0.80g/L according to the Chilean Liquid Chromatography. The mobile phase was legislation). In 25% of the cases the presence of drugs acetonitrile:ammonium acetate. Blood and urine was detected and the average of these was 1.78 samples were screened with Chemiluminescence drugs/case, whereas in 16.1% of the suicides detected Immunoassay. Confirmatory assay by the presence of alcohol and drugs together. The most Chromatography tandem Mass Spectrometry. All frequent groups of drugs that were detected were results were plotted in tables and presented in cocaine, benzodiazepines and marijuana. percentage and absolute values. Results: 100% of cases had hair samples for analysis, approximately 91% had blood and only 59% of them had urine. For the blood FO 03 - CHARACTERIZATION OF THE samples, in only 35% of cases COC was detected. In SUICIDE POPULATION IN AUTOPSIES urine samples this value decreases, being detected in PERFORMED IN THE REGION OF ATACAMA - only 23% of them. These percentage increased in hair CHILE, 2007 TO 2016 to 68% of the analyzed samples. The concentration ranges of 2.17 to 127.05 ng/mg of hair. There were 2 Cruz, C. E.; Espinoza C.M. 1, Araneda E.C. 1, Silva cases that don’t have neither blood nor urine collected. C.A. 2, Novakovic I.M. 2, Flores D.R.1. In these cases, the hair analysis result shows COC detection. Besides, there were 3 cases that were 1. Laboratory of Forensic Medical Service of Copiapó, undetected for COC on blood and urine but detected in Chile. 2. Unit of Thanatology of Forensic Medical hair. Conclusion: The hair is a good option as an Service of Copiapó, Chile. alternative sample, considering that it was available in 100% of cases. It was the sample that presented the The World Health Organization in the last report, highest percentages of COC detection. It also had a informed that the death rate for suicides in the world larger detection window than blood and urine. Finally, was 10.7 per 100 000 population by 2015 and a 1.7:1 the use of this alternative sample is important for the male:female relation; according to the same source, in forensic toxicology. Chile this rate was 9.9 and a relation of 4.7:1 male:female that same year. The objective of the present study was to characterize the suicidal ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 140 population according to the autopsies performed in the material on doping, alcohol use and illegal drugs, Atacama region - Chile, between 2007 and 2016. A informing the public about the social and health descriptive-retrospective study of the last 10 years was implications of use of drugs, explain the chemical carried out for the cases of death with etiology suicide. properties and forensic aspects. Materials and The variables analyzed were age, sex, marital status, methods: Bibliographic review; Preparation of method of suicide, place of occurrence, toxicology and presentations in slides containing the main subjects to other. Of 1637 autopsies studied, 20.1% corresponded be approached, will be chosen according to the domain to suicide deaths (n=330), resulting in an average of the subject by the target audience; Preparation of regional rate of 11.1 per 100 000 population; it questionnaires that will be applied before and after highlights the year 2011, with a rate of 16.2. Related to each presentation; Presentation of the presentations gender, the male: female relation was 4.9:1 and the according to the availability and interest of the hanging method was used in 87% of the cases. The community. Result and conclusion: In collaboration presence of alcohol was detected in 45.8% of the with the Extension Program " Desvendando as autopsies of suicidal etiology and drugs of abuse in Ciências Forenses", the presentations were made to 25%, with preponderance of Cocaine. Based on these elementary and middle school students from Manaus. results, the present study concluded that the average The experience was positive, the studantes showed regional suicide rate is higher at the national and world great interest in the subject addressed, 92% of students rates, and the male:female relation is similar to the consider it important to learn about forensic national rate and almost three times higher than the toxicology, and 96% felt that speeches helped in world rate. understanding the issues. Financial support / Acknowledgments: PROEXT - UFAM, Universidade Federal do Amazonas, Programa FO 04 - FORENSICS TOXICOLOGY: DOPING, de Extensão Desvendando Ciências Forenses. ALCOHOL, ILLEGAL DRUGS

SILVA, E. S. 1; NÓBREGA, L. C. 2; HORA, L. F. 3; FO 05 - CASE STUDY: FATAL INTOXICATION BENTES, K. R. S. 4; LUCAS, A.C.S. 5. WITH POISONED SYRUP

1 Universidade Federal do Amazonas, Manaus - AM, LEITE, F. P. 1; GOLVEIA, J. C. S. 1; COLLETES, T. Brasil; 2. Universidade Federal do Amazonas, Manaus 2; LEITE, B. H. M. 1; RIBEIRO, R. 1; - AM, Brasil; 3. Universidade Federal do Amazonas, ZANCANARO, I. 1; BRAGA, P. C. C. S. 1; VAZ, B. Manaus - AM, Brasil; 4. Universidade Federal do G. 2; LOBO, S. W. Amazonas, Manaus - AM, Brasil; 5. Universidade Federal do Amazonas, Manaus - AM, Brasil. 1 Polícia Técnico Científica de Goiás, Goiânia - GO; 2 Universidade Federal de Goiás, Goiânia – GO. Abstract: The forensics toxicology is a multidisciplinary science that acts directly in law, Introduction: On 06/18/2017, the Child and Adolescent International and Brazilian legislation is full of Protection Police Unit (DPCA/GO) assigned the regulations about some chemical substances that can Technical-Scientific Police Superintendence of Goiás be analyzed based on analytical toxicology, helping to State (SPTC/GO) to analyze a Brondilat® syrup flask clarify the presence in biological matrices and (Acebrophilin 50 mg/5 mL, 120 mL, Aché laboratory) possibles toxic effects. In sports, a list of banned related to a two-year-old female. The case history was: substances among athletes, since they could benefit the "on 06/17/2017, after the ingestion of a dose of the performance artificially, which constituted ‘‘doping’’. medicine, the minor felt sick, underwent cardiac arrest The psychoactive drugs, in Brazilian legislation and is currently hospitalized in a grave condition at defines those that are legal and illegal drugs, in the HUGOL". In addition, biological specimens analysis country the drug abuse of alcohol, cocaine, marijuana was requested. Promptly, Forensic Experts of the remains a problem of public health, social, economy Toxicology Unit (SETOX) of SPTC/GO attended the and legal. For alcohol, there are restrictions of use as Intensive Care Unit of the Emergency Hospital between drivers, due to the serious potential Governor Otávio Lage de Siqueira (HUGOL) in consequences for user and for public security. With Goiânia/GO, for the victim’s blood and urine regard to illegal drugs, use is not allowed and there is collection. The genitor authorized the procedure extensive legislation regarding the penalties for the use throughout a Collection Term. In addition, the forensic and trafficking of these substances. Thus, with the experts applied a questionnaire aiming the collection realization of the project, it is expected to make the of useful data to the analysis. Objective: The work population aware of the effects of these substances on aimed the toxicological analyses on the seized syrup the life and health of each individual, serving as and the victim’s specimens and the comparison of their prevention of their use. Objective: Elaborate didactic contents. Material and Methods: In order to perform ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 141 the toxicological analyses, part of the exams was ammonium formate 2 mM and 0,1% of formic acid in proceeded at SETOX, using a gas chromatograph (A) water (B) methanol in a 16 minute run. Linearity coupled to mass spectrometer (GC-MS). Another part was obtained ranging from 25 (LOQ) to 1,000 ng/mL was at the Laboratory of Chromatography and Mass for the four compounds (r > 0.99, 1/x weighted linear Spectrometry (LaCEM) located at the Institute of regression). Validation for this method is following the Chemistry, Federal University of Goiás (UFG), using a recommendations from Scientific Working Group for mass spectrometer by direct infusion in paper spray Forensic Toxicology (SWGTOX) for quantitative ionization - with ion trap analyzer (PSI / ion trap) and analysis. Precision (CV (%)) and bias (%) were high resolution mass spectrometry by direct infusion evaluated within and between runs at 3 different levels with ion trap-orbitrap analyzer. Results and (75, 400 and 800 ng/mL), never exceeding 20% at Conclusion: GC-MS analysis of the questioned flask each concentration. Matrix effect and dilution integrity sample and blood and urine samples indicated the precision and bias criteria were also tested and met. presence of methomyl (C5H10O2N2S) - insecticide Stability is being study for a period of 0,7,15,30,60 and and acaricide of specific agricultural pertaining to the 90 days. Since this method is so simple and the use of class of carbamates, classified as extremely toxic dried sample has so many advantages, it is expected (ANVISA, Brazil). The result was confirmed using the that in the future it can be used in forensic toxicology PSI/ion trap and by high-resolution mass spectrometry routine. analysis. Therefore, the content in the flask was either Financial support / Acknowledgments: CAPES adulterated or replaced by the pesticide aforementioned, resulting in the victim’s death. Financial support / Acknowledgments: FAPEG FO 07 - VITREOUS FLUID: ANALYSIS OF OPIATES USING EXACTIVE FTMS

FO 06 - LIQUID CHROMATOGRAPHY– SANTOS-JÚNIOR, J. C.; MESQUITA S. 3; MOLLO TANDEM MASS SPECTROMETRY APPLIED FILHO, P. C. 4; TARTARELLA M. A. 4; TO THE STUDY OF COCAINE AND VENDRAMINI P. H. 1; EBERLIN, M. N. 1; HÖEHR, METABOLITES IN DRIED URINE SPOTS (DUS) N. F 2.

BARBOSA, I. L.; COSTA, J.L 1; EBERLIN, M. N. 2. 1 Thomson Mass Spectrometry Laboratory. Institute of Chemistry, UNICAMP, Brazil; 2 Department of Universidade Estadual de Campinas. Clinical Pathology, School of Medical Sciences, UNICAMP, Brazil; 3 Clinical Biochemistry Dried blood spots (DBS) provides several advantages Laboratory, School of Medical Sciences, UNICAMP, over conventional sample storage methods, mainly in Brazil; 4 Team of Forensic Medicine West. Medico- what it concerns transport and stability of the analytes Legal Institute, Police Technical Scientific and it has been used for clinical and forensic Superintendence — SPTC, Brazil. applications. Similarly, it is possible to perform toxicological analysis of drugs of abuse in samples of Introduction: The term “opiates” refers to substances dried urine spots (DUS) and it should be of interest derived from the opium poppy, such as morphine and supposing that shares the same advantages as DBS. In also includes some semi-synthetic substances like this work, a very simple analytical method for DUS heroin.[1] Vitreous fluid is composed of 99% water, and LC-MS/MS was developed for the screening and with the remaining 1% made up of sugar, salts and confirmation of cocaine and its metabolites: proteins. It is also subjected to less contamination and benzoylecgonine (BZE), ecgonine methyl ester (EME) bacterial degradation due to the protected environment and cocaethylene (EC). To prepare DUS cards, spiked inside the eye, which makes it available in cases in urine samples (20 µL) were spotted onto Whatman 903 which blood samples have already been degraded.[2] Protein Saver cards followed by 2 hours air-drying Exactive ESI FTMS technique, provides a one-to-one prior to extraction. The extraction was performed using molecular formula m/z relationship due to its ultra high 300 µL of methanol:water (60:40) with 0,1% formic resolution and mass accuracy.[3] Integrating acid, containing the internal standard (cocaine-d3, 10 toxicological scopes, the aim of this work was ppb) followed by 15 minutes of vortex and 5 minutes therefore to develop a method based on Exactive of centrifugation at 10.000 rpm. After this process, 200 Fourier Transform Mass Spectrometry (Exactive µL of supernatant was transferred to a vial and 15 μL FTMS) for analysis of opiates and their metabolites in injected on a UPLC-MS/MS system (8040 Shimadzu, vitreous fluid non-decomposing bodies. Method: The Japan). Separation was performed in a C18 column samples were obtained from 10 bodies attended by (Atlantis T3, 150 x 3 mm, 3 μm, Waters®) with a team of forensic medicine from the West zone from gradient elution with a flow rate of 0.40 mL/min. the Medico-Legal Institute of Police Technical Chromatography separation was carried out using Scientific Superintendence of São Paulo, Brazil. This ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 142 study was approved by the Research Ethics identifying users profile in the city of Brasília through Committees (Ethics Protocol Approval numbers sewage analysis employing solid phase extraction- 1270/2010, 09/11, 687/2012 and 736/2012). The liquid chromatography-tandem quadrupole-time of samples were composed of 100 µL of vitreous fluid flight mass spectrometry (SPE-LC-QTOF).Material (diluted 100x) and added 50 µL of 150 ng/ml opiates and Methods: Composite sewage samples were (heroin and morphine) and their metabolites solutions collected in two treatment plants, namely north-wing both diluted in 1 mL of acetonitrile with 100 mM (NW-STP) and south-wing (SW-STP). Aliquots of 50 Ammonium Acetate. The mass/charge (m/z) of opiates of mL (pH 2) were passed through SPE cartridges were monitored in ESI positive mode by a Q Exactive (StrataX) conditioned with MeOH:ACN (60:40) and hybrid quadrupole-Orbitrap mass spectrometer water (pH 2). Cartridges were frozen and taken to (Thermo Fisher Scientific). These drugs and their Spain where the analytes were recovered with MeOH. metabolites were identified by comparison with the Extracts were spiked with deuterated standards and mass spectra of compounds standard. Results and concentrated to 0.2 mL in a TurboVap® II Evaporator. conclusion: In the present work, opiates and their Analyses were carried out using an Agilent 6520 metabolites were detected in post-mortem vitreous equipped with a dual electrospray ionization. Results fluid analyzed. Thus, for its analytical stability and and Discussion: The area covered by the NW-STP sterility after death, vitreous fluid is a good sample for presented average cocaine consumption of 8,01 and toxicological analyzes. Finally, the proposed method 4,01 doses inhab-1 y-1 in weekends and non-weekend based on analysis of opiates by Exactive FTMS proved days, respectively. Analogously, analysis of the SW- to be satisfactory, constitutes an important tool for STP samples revealed a consumption of 4,13 and 1,33 "fingerprinting" in the post-mortem toxicological doses inhab-1 y-1. The ratio CE/BE was higher on analysis, which is essential to the forensic toxicology Saturdays, confirming that cocaine is commonly routine. consumed with alcohol during weekends. Conclusion: Financial support / Acknowledgments: The authors The method presented here was accurate for the are grateful to the Conselho Nacional de analysis of cocaine and metabolites in sewage samples. Desenvolvimento Científico e Tecnológico (CNPq) for The QTOF system allows the unambiguous the scholarship (J. C. S. J.) and fellowship (M. N. E) identification of substances based on accurate mass and to the Fundação de Amparo a Pesquisa do Estado measurements and matched isotope patterns. Our de São Paulo (FAPESP) for sponsoring this research. results revealed that intermittent cocaine users may be an important group to consider in further prevention and educational actions. FO 08 - QUANTITATIVE DETERMINATION OF Financial support / Acknowledgments: COCAETHYLENE, COCAINE AND FAPDF BENZOYLECGONINE BY LC-MS/MS TO ESTIMATE CONSUMPTION AND INDENTIFY USER'S PROFILE FO 09 - DEVELOPMENT AND VALIDATION OF THE FIRST METHOD TO ANALYSIS OF SILVA, K.M. 1; QUINTANA,J.B 2; GONZALEZ,I.M NBOMES IN DRIED BLOOD SPOT (DBS) 2; RODIL, R.2; BATISTA, L.F.3; SODRÉ, F.F.1. CUNHA, K. F. 1; COSTA, J. L. 1,2. 1 University of Brasília, Chemistry Institute; 2 University of Santiago de Compostela, Institute of 1. Centro de Informação e Assistência Toxicológica de Food Analysis and Research; 3 Companhia de Águas e Campinas, Faculdade de Ciências Médicas, Esgotos de Brasília (CAESB). Universidade Estadual de Campinas, Campinas - SP, Brasil; 2. Faculdade de Ciências Farmacêuticas, Introduction: The monitoring of drug use by sewage Universidade Estadual de Campinas, Campinas - SP, analysis is a topic of emerging concern. Previous Brasil. results by our group show an increase in cocaine use at weekends. However, one can consider the existence of Introduction and objective: NBOMes, N- two distinct groups of users, e.g., heavy chemical benzylmethoxy derivates of the 2C hallucinogens dependents and intermittent users, who make social family, have been used as a legal alternative to lysergic use of a drug. Consumption estimates based on the acid diethylamide (LSD). A liquid chromatography- levels of the major cocaine metabolite tandem mass spectrometry (LC-MS/MS) method has (benzoylecgonine-BE) are not able to differentiate the been developed and validated for the quantitative user profile. However, some strategies involve the co- analysis of seven NBOMes (25C, 25H, 25I, 25B, 25G, quantification of cocaethylene-CE, a product of the 25D and 25E-NBOMe), in human dried blood spot metabolism of cocaine with ethanol. Objective: The (DBS) samples. Method: For method development and goal of this work was to estimate drug consumption validation, spiked whole blood (15 μL) was applied in ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 143

6 mm disks, previous picked from Whatman 903 Protein Saver cards. DBS disks were dried for 3 hours Introduction: use of cocaine and other drugs is at room temperature and extracted with 300 μL of associated with trauma, representing a problem in methanol-acetonitrile mixture (75:25, v/v), containing emergency care. Toxicological analyzes (TA) for the internal standard (LSD-d3, 0.5 ng/mL). This detection of drug use are fundamental in these mixture was vortexed for 10 min and centrifuged at services. TAs include screening by rapid kits and 10.000 rpm for 5 min, and 10 μL injected on LC- confirmation by gas chromatography-mass MS/MS (Agilent 1260 Infinity coupled to ABSciex spectrometry (GC-MS). Aims: to validate an analytical 5500QTRAP). Separation was performed in C18 method for identification of cocaine and its column (Atlantis T3, 150 x 3 mm, 3 μm, Waters®) and biotransformation products in urine by GC-MS. gradient elution (acetonitrile-water with 0.1% formic Methods: analytical validation process followed acid) with a flow rate of 0.5 mL/min. Mass Resolution No. 899, May 29, 2003, ANVISA. spectrometer was set operate in multiple reaction Negative samples, obtained from volunteers, were monitoring mode (positive electrospray ionization). enriched with cocaine (COC) and benzoylecgonine Results: During the method development, we decide to (BZE) and internal standards COC-deuterated and use fixed blood volume and whole spot analysis, in BZE-deuterated. Urine samples were extracted by order to minimize the hematocrit effect. Method limits mixed-phase extraction cartridges, Agilent Bond of detection (LOD) and quantitation (LOQ) were 0.05 ElutCertify 130mg/3mL. Extracts were resuspended in and 0.1 ng/mL, respectively. Linearity was obtained 25μl of N, O-Bis (trimethylsilyl) trifluoroacetamide from 0.1 to 10 ng/mL for all NBOMes (r > 0.99, 1/x2 (BSTFA) (1% trichloromethylsilane) and 25μl ethyl weighted linear regression). The precision (within-run acetate, derivatized at 90°C/15min. in heating plate and between-run, % RSD) and bias (%) were evaluated and analyzed by GC-MS Thermo Fischer Scientific, at 0.3, 2 and 8 ng/mL, and weren’t higher than 18%. CG Focus and EM DSQ II, equipped with No interference was observed when LOQ was InjectorTriplus AS, HP-5MS silica capillary column analyzed in presence of 500 ng/mL of other drugs of (30m x 0.25μm x 0.1μm).The method has 17 min, abuse and pharmaceuticals. The matrix effect varied starting at 160°C and holding for 2 min, with a heating between 50.5% for low QC and 64.6% for high QC. ramp up to 300°C at 20°C min-1 and Helium flux of The stability was studied at -20ºC, 4ºC and room 0.6 mL.min-1. Injection was done in splitless mode. temperature during 180 days; the decrease of the The mass spectrometer operated at 250°C in the concentration wasn’t higher than ±20% during this electron impact mode with energy of 70 eV. The period. Conclusion: This is the first validated method quantification was performed in selective ion to quantify NBOMes in DBS. Even small quantity of monitoring mode. Results: COC and BZE were sample was enough to reach a high sensitivity, which confirmed by identification of their reference mass is required for these analytes. spectrum. Method was linear over the specified range Financial support / Acknowledgments: FAPESP, 15 to 2000ng/mL; r2 0.9985, for COC and 20 to FAEPEX, CNPq 3000ng/mL; r2: 0.9999 for BZE. The method presented precision ranging from 5.4-14.6% and 7.8- 12.3%, for COC and BZE, respectively. Recovery of FO 10 - VALIDATION OF AN ANALYTICAL the analytes was in the range 93.5-102.1% and 97.5- PROCEDURE FOR DETERMINATION OF 104.8% for COC and BZE, respectively. The limits of COCAINE AND BIOTRANSFORMATION detection and quantification were 15 and 10ng/mL, 20 PRODUCTS AS DIAGNOSTIC SUPPORT IN and 15ng/mL, respectively for COC and BZE. THE CONFIRMATION OF USE OF DRUGS IN Conclusion:the validated intra-laboratory method is TOXICOLOGICAL EMERGENCY CARE simple and showed good precision and accuracy. The procedure proved to be effective, safe and applicable OLIVEIRA, K. O.; MENOTTI, V.S.1; SCANFERLA, in Emergency Care to evaluate the use of cocaine in D.T.P.2; FRATUCCI, G.F.2; ALVES, G.S.2; urine. BANDO, E.2; NERILO, S.B.2,3; MACHINSKI, Financial support / Acknowledgments: M.J.2; OLIVEIRA, M.L.F4; MOSSINI, S.A.G1. Decit/SCTIE/MS; CNPq; Araucaria Foundation and SESA-PR for their support. 1.Postgraduate Program on Bioscience and Physiopathology, State University of Maringá, Maringá – PR, Brazil; 2. Laboratory of Toxicology, Department of Basic Health Sciences, State University of Maringá, Maringá – PR, Brazil; 3. University Center Ingá, Maringá – PR, Brazil; 4.Department of Nursing, Postgraduate Program in Nursing, State University of Maringá, Maringá – PR, Brazil. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 144

FO 11 - POSITIVITY FOR DRUG USE IN alcohol. The positive samples were confirmed by GC- PHYSICAL TRAUMA - COMPARISON OF MS. Between the negative samples with positive blood RESULTS BY SCREENING AND alcohol, 5 samples were positive for COC and/or BZE CONFIRMATORY TESTS by GC-EM, with values higher than 150ng/mL, evidencing the efficiency and importance of the OLIVEIRA, K.O.1; SILVA, M.2; MENOTTI, V.S.1; confirmatory test. Conclusion: Results evidenced the KIMOTO, K.Y.3; OLIVEIRA, N.G.3; SIQUEIRA, effectiveness of the confirmatory test and the A.P.C.S.3; ALTAFINI, D.D.4; BANDO, E.3; importance of TAs as diagnostic support in Emergency MACHINSKI, M.J.3; OLIVEIRA, M.L.F.2,5; Care. MOSSINI, S.A.G.1,3. Financial support / Acknowledgments: Decit/SCTIE/MS; CNPq; Araucaria Foundation and 1.Postgraduate Program on Bioscience and SESA-PR for their support. Physiopathology, State University of Maringá, Maringá – PR, Brazil; 2.Postgraduate Program in Nursing, State University of Maringá, Maringá – PR, FO 12 - FLUNITRAZEPAM: THE USE OF THE Brazil; 3.Laboratory of Toxicology, Department of DRUG AS RAPE DRUG Basic Health Sciences, State University of Maringá, Maringá – PR, Brazil; 4.Laboratory of Clinical VICTÓRIO, P. C. 1; MARCELLINO, A. E.O. 1; Analysis, Regional University Hospital of Maringá, FIGUEREDO, R. M. 1. Maringá-PR, Brazil; 5.Intoxication Control Center, Regional University Hospital of Maringá; Maringá – 1 Universidade Católica de Santos, Santos-SP, Brasil. PR, Brazil. INTRODUCTION: In the year 1975, flunitrazepam Introduction: Toxicological analyzes (TA) for was released by the pharmaceutical industry. This drug detection of drugs and their biotransformation products belongs to the class of benzodiazepines, drugs are fundamental in Emergency Care. Use of cocaine depressants of the central nervous system, which are and other drugs are associated with physical trauma, used as hypnotics, anxiolytics, anticonvulsants and representing a problem in these services. TAs for use muscle relaxants. However were reported cases of its detection include screening by rapid kits and improper use as rape drug, because when added to confirmation by Gas Chromatography-Mass drinks and because of does not have color, smell or Spectrometry (GC/MS), a high sensitivity and taste, is imperceptible to the victim. It is common than specificity technique. Cut off values of 300 ng/mL and the drug can be added on alcoholic drinks worsening 150 ng/mlL, for detection of benzoylecgonine (BZE), the condition of the victim, because this drug has its the main biotransformation product of cocaine in urine, effect potentiated by interaction with other drugs are recommended by international guidelines for depressing, especially the alcohol. Due to abusive and screening and confirmatory test, respectively. Aims: indiscriminate use, in 1983, the drug was added to the To compare results by toxicological screening with group of controlled substances in the United States of confirmatory test in samples from victims of trauma America and its sale was banned. In France, the coup, associated with drug use treated in an Emergency better known as "chemical submission", also has done Care. Methods: The study was carried out in an many victims and is difficulty for official records.For Emergency Care in the Northwest of Paraná. the hospital authorities, there are case of repeat Population study was selected by epidemiological offenders, some of whom have recorded two or three surveillance of sentinel events, identified by diagnosis overdoses and most of them with "euphoric amnesia". of physical trauma associated with drug use from 2015 OBJECTIVE: The objective was to analyze the data and 2016. Biological samples obtained from the notified of rape within the national and international individuals under study were used to TAs. Positive levels in order to try to differentiate the cases on which samples in screening by rapid kits and negative ones, it is possible to identify that the drug had been used. but positive for blood alcohol content, were analyzed MATERIAL AND METHODS: The research method by GC-EM (Thermo Fischer Scientific), equipped with was based on scientific articles, data of Public Safety GC gas chromatograph, DSQ II Mass Spectrometer, and the World Health Organization. RESULTS: Injector Triplus AS, HP-5MS silica capillary column According with UN, in the world, 12 million women (30m long, 0.25μm diameter and 0.1μm film). The are raped each year. Data from Department of Public research project was approved by Committee of Ethics Security of São Paulo show that, during the period in Research Involving Human Beings of the State from January to September/2016, in São Paulo, there University of Maringá, opinion 458.185. Results: were 1668 cases of rape. Whereas, less than 1% are There were carried out screening by rapid test for 205 associated with Flunitrazepam and only 10% of all samples from victims of trauma, 38 were positive for crimes of rape are reported to police. CONCLUSION: cocaine and 16 negative, but with positive blood It is imperative to have the empowerment of victims to ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 145 denounce, greater rigor in the protocol of sexual abuse presented values from 97.6 to 114.0%. The proposed and mandatory biological research of Flunitrazepam as method is a promising alternative to fast extraction and a facilitator to rape. analysis for avoiding laborious process, and for Financial support / Acknowledgments: Course of showing precision and accuracy according to the Pharmacy and Institute of Research Science and values proposed by ANVISA. Technological of Universidade Católica de Santos and Financial support / Acknowledgments: CAPES, Military Police of São Paulo. FAPEG

FO 13 - A NEW METHOD FOR DIRECT DRUG ANALYSIS BASED ON PROBE ELECTROSPRAY IONIZATION MASS SPECTROMETRY

BERNARDO, R. A. 1; COLLETES, T. C. 1; VAZ, B. G. 1; CHAVES, A. R. 1.

1 - Universidade Federal de Goiás, Goiânia - GO, Brasil.

The Probe Electrospray Ionization Mass Spectrometry (PESI-MS), first reported in 2007, consists of a metallic probe is previously inserted in a sample solution and then directed coupled to mass spectrometer for desorption and analysis. In this study, a solid phase microextraction (SPME) based on conductive polymer (polypirrole) was developed and applied as PESI-MS method for drugs (Cocaine, Lisergic Acid Diethylamide (LSD), Methamphetamine (MA) and [3,4 – Methylenedioxymethamphetamine] (MDMA)) in biological fluids analysis. A platinium wire was used as work electrode for electrodeposition of Polypyrrole (PPy) films by cyclic voltammetry technique. The coating was electrodeposited by cyclic voltammetry using potential range from 0 to 1.2 V at scan rate of 50 mV s-1. The developed probe was inserted in urine and oral fluid samples spiked with standard solutions of cocaine, LSD, MA and MDMA in a concentration range from 1 to 500 ppb. The extraction variables such as: time, ionic strength and pH was evaluated. For desorption process, the probe was positioned at 5-7mm from the mass spectrometer inlet. A high voltage (3 kV) was applied to generate the PESI (+) mass spectra. The electrodeposition parameters were optimized and a better extraction extraction was obtained with 5 voltammetric cycles. The drug quantification in oral fluid and urine samples was performed by MS tandem experiments, where the fragment ion intensity was used to build the analytical curves. According to the results of analytical essays, the coated PESI-MS method presented linearity in a concentration range from the 2 (limit of quantification, LOQ) to 500 ng.L-1 (ppb). The LOQ values ranged from 2 ppb to 10 ppb. The interday precision presented values from 0.8 to 9.6% with accuracy ranging from - 0.6 to 13.7%. The intraday precision values varied from 1.7 to 10.8% with accuracy ranging from -2.4 to 14.0%. The absolute recoveries were evaluated and

GENOMICS, PROTEOMICS AND METABOLOMICS ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 147

OM 01 - A METABOLOMICS STUDY OF OM 02 - MAOA VNTR POLYMORPHISM AND GESTATIONAL EXPOSURE TO SMOKE OF AGE AT SMOKING INITIATION IN Cannabis sativa IN MICE BRAZILIAN MALES

SOUZA, A.R.L.1; FERNANDES, A.P.G.1; SILVA, MOTA, C.L. 1; MITRI, S. 1; BARATA-SILVA, C. 1; N.S. 1; TORRES, J.I. 1; DÖRR, F. 1; PINTO, E. 1; PAVESI, T. 1; MOREIRA, J.C. 1 VERAS, M.M. 1; TAVARES, M.F.M. 1 1 Escola Nacional de Saúde Pública Sergio Arouca 1 Universidade de São Paulo, São Paulo-SP, Brasil. Introduction: Smoking is a public health problem and Introduction: Metabolomics has been applied in is considered the leading cause of preventable death in Toxicology to reveal the mechanisms of action of the world. Over 5300 compounds have been identified xenobiotics and in the discovery of biomarkers. in tobacco smoke, and over 70 carcinogens with Cannabis sativa L. is one of the most commonly sufficient evidence for carcinogenicity. The nicotine is consumed illicit drugs among pregnant women the major psychoactive substance in tobacco, and resulting in teratogenic and neuro-behavioral effects. stimulates the release of dopamine, the key After drug smoking, delta-9-tetrahydrocannabinol (Δ9- neurotransmitter in nicotine dependence, leading to THC), the major cannabinoid of Cannabis, may feelings of pleasure and reward. The addictive nature overcome placental and blood-brain barriers. The of nicotine is multifactorial, involving both substance is metabolized in the liver, producing 11- environmental and genetic factors. Birth decade, low nor-9-carboxy-tetrahydrocannabinol (THC-COOH). socio-economic status, low education level, alcohol Objectives: To evaluate the impact of gestational consumption, and beginning smoking during exposure to Cannabis sativa smoke in the brain of adolescence have been found to be environmental risk offspring mice as well as to determine THC-COOH in factors for tobacco smoking. Genetic factors also affect the urine of exposed animals. Material and Methods: smoking behavior. In fact, genetic variation may Pregnant females (2) were exposed to daily doses of influence specific elements of smoking status. Cannabis smoke inhaled for 5 min (200 mg drug with Estimates indicate that genetic variation may account 0.3% Δ9-THC) from day 5th to 17.5th of gestation. for 44–56% of smoking initiation. Thus, genes that After birth and weaning, a group of offsprings (10) regulate the concentration of dopamine can influence was re-exposed for another 60 days. The animals were the smoking behavior, as the gene encoding the MAO divided into exposed to Cannabis sativa smoke (test enzyme responsible for the dopamine degradation. In group, n=5) and exposed to filtered air (control group, the present study, we investigated the relationship n=5). Urine was collected after 24h exposure. Alkaline between a polymorphism in the promoter region of the hydrolysis followed by liquid-liquid extraction and MAOA gene and age at smoking initiation in Brazilian derivatization (o-methoxyamine in pyridine for males. Methods: This is a cross-sectional study with methoximation and BSTFA + 1% TMCS for the participation of 121 men. The polymorphism was silylation) were performed for gas chromatography- determined in the sample population by the technique mass spectrometry (GC-MS) analysis of THC-COOH of Polymerase Chain Reaction (PCR). Results and in urine. Brain samples were extracted with 1:1 Conclusions: In this population study, the age of methanol:water and derivatized prior to GC-MS smoking initiation was 16 years old, considered the metabolomics analysis. For data processing, XCMS, period of greatest risk for the beginning of smoking AMDIS and SIMCA P+ softwares were used. Results: habit. The main motivation for the initiation of Concentrations of THC-COOH in the mice urine smoking was influenced by friends. Statistical analysis ranged from 4.4 to 30 ng/mL confirming gestational revealed no significant association between the exposure. In the metabolomics study, the discriminant polymorphism MAOA-u VNTR and the age at metabolites, glycolic acid, palmitoleic acid and urea smoking initiation. Given the importance of the issue, were decreased in the test samples whereas caprolic studies with larger sample sizes will be needed to acid, D-malic acid, dehydroascorbic acid, pyruvic acid, contribute to the elucidation of the genetic mechanisms D-lyxose, glycerol 1-phosphate were increased. and tobacco susceptibility. Conclusion: The results suggested that gestational Financial support/Acknowledgments: Capes, exposure to Cannabis sativa has a measurable impact Fiocruz. on the metabolism of Δ9-THC in the offsprings brain and may lead to a better understanding of chronic exposure and fetal development. Financial support/Acknowledgments: CAPES/CNPq

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OM 03 - RNA-SEQ-BASED TOXICOGENOMICS Introduction: Quercetin is a flavonoid available in COMPARISON OF AIRWAY CELLS EXPOSED different plants. Studies have shown the generation of TO VypeTM ePEN VAPOUR AND CIGARETTE metabolites of this compound, among them SMOKE glycosylated derivatives that present considerable cytotoxicity. Biotransformation reactions are used in MINET, E. 1; HASWELL, L. 1; BAXTER, A. 1; the structural modification of a compound to a specific BANERJEE, A. 1; MUSHONGANONO, J. 1; product by biological catalysts such as filamentous ADAMSON, J. 1; THORNE, D. 1; GACA, M. 1. fungi. Understanding the metabolic pathway used by the microorganism for structural modification of 1. British American Tobacco, R&D Centre, substrate presents as an important tool that can aid in Southampton SO15 8TL, United Kingdon. the mechanisms of control for its induction or inhibition. Aim: The aim was to know the pathways of Introduction: VypeTM ePen can be used by cigarette reduction and glycosylation of quercetin using smokers as a substitute of smoking. There is increasing descriptive proteomics analyzes. Methods: B. bassiana evidence from in vitro testing that such products cause was incubated in 100 ml of Czapek. The spores of minimal damage to cell systems, however there are fungus were added, kept at 28 °C and 200 rpm. After few studies reporting cytotoxicity and inflammatory 65 h of growth, was added quercetin (25 mg/ml) until responses. These discrepancies arise from the use of 120 hours. After, the fungal pellets were separated and different cell models, products, exposure matrices, washed with 200 μM ammonium bicarbonate and times and doses. Ideally, a comprehensive in vitro extractions process of proteins was followed by assessment of vape products should use a relevant cell maceration with liquid nitrogen. Enzymatic digestion model and well-considered exposure strategy. of the proteins was performed with trypsin and the Objective: We compared the transcriptional response tryptic peptides, separated by high performance liquid of a primary 3D airway model (MucilAirTM) exposed chromatography at nanoscale using the ACQUITY for one hour to VypeTM ePen vapour and smoke from UPLC® M-Class system and processed in the a reference combustible cigarette (3R4F). The ProteinLynx Global Server 3.0.2 program. Results: originality of this work is a careful consideration for The production of enzymes involved in reduction and the dose where instead of matching the exposure glycosylation reactions was observed. The reduction between different products based on puff number, we reaction of the carbonyl of quercetin was catalyzed by used nicotine as a surrogate for dose matching since an alcohol dehydrogenase. The hydroxyl resulting smokers titrate for nicotine. Methods & Results: The from this reaction may have acted as an acceptor group average nicotine delivered to the cells from 3R4F on the glycosylation reaction. Sucrose from the culture smoke at a 1:30 dilution was matched at a dilution of medium was hydrolyzed by α-glycosidase and then 1:7 for the e-cigarette. One additional e-cigarette undergoing adenosine kinase action, forming a dilution of 1:3 was used also tested. RNA was glucose-6-phosphate, or phosphotransferases, forming extracted for RNA-seq from the tissues at 24hrs and a glucose-1-phosphate. A 1,3β-glucanosyltransferase 48hrs post exposure. 873 and 205 RNA features were probably performed the catalysis of transglycosylation differentially expressed for 3R4F smoke at 24hrs and reactions between quercetin and sugar. Another 48hrs post exposure using a pFDR2 threshold, probable route is by glucan 1,3-beta-glucosidase. respectively. Differentially expressed RNA from e- Conclusion: The proteomic analyzes allowed the cigarettes (49 RNA at 1:7 dilution and 113 RNA at 1:3 knowledge of the likely ways of reduction and dilution) could only be identified using a looser glycosylation of quercetin, assisting in studies to threshold of pFDR. prevent the formation of these metabolites. Financial support/Acknowledgments: FAPEG.

OM 04 - PROTEOMICS REVEALS A PATHWAY FROM ENDOGENOUS PRODUCTIONS OF OM 05 - IDENTIFICATION OF DELETIONS CITOTOXIC QUERCETIN METABOLITE AND DUPLICATIONS IN THE CYP2C9 GENE IN A POPULATION IN GOIÂNIA – GO BY ARRUDA, E.L. 1; JAPIASSU K. B. 1; SOUZA, P. L. MLPA. M. 1; CARVALHO, P. H. O. 1; BAEZA, L. C.2; SOARES, C. M. A.2; OLIVEIRA, V. 1 TEIXEIRA, T.M.¹; PEREIRA L C G¹; BERGAMO N A¹; SILVEIRA-LACERDA E P¹ 1. Laboratório de Bioconversão, Universidade Federal de Goiás,Goiânia -GO, Brasil; 2. Laboratório de 1- UNIVERSIDADE FEDERAL DE GOIAS, GO, Biologia Molecular, Universidade Federal de BRASIL Goiás,Goiânia -GO, Brasil ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 149

Metabolic human genes of the Cytochrome P450 toxic elements promote alterations in protein synthesis (CYPS) family are important in physiology, drug that may be linked to the mechanisms of toxicity. metabolism, and pathogenesis of certain diseases, and Then, the present study aimed to investigate there is a strong need to explore the genetic variations modifications in the plasma proteome of subjects and regulatory mechanisms of these enzymes. occupationally exposed to Pb. Materials and Methods: Mutations in CYP genes leading to deficiency in the Blood and plasma samples were obtained from: i) male enzymes result in a wide spectrum of human diseases. battery workers (n=5) and ii) a group with no history Cytochrome P450 2C9 (CYP2C9) is an important of occupational exposure to Pb (n=5), in Paraná State, enzyme in the metabolism of many drugs, including NSAIDs, antidepressants and anticoagulants. The aim Brazil. All study participants answered a questionnaire of the present study was to investigate, the importance to provide information on life-style, exposure time, of CYP2C9 genetic polymorphisms in the health and health histories. Blood lead levels (BLL) were population in Goiânia. The study was performed determined by inductively coupled plasma mass acording approved by the ethics committee of Federal spectrometry (ICP-MS). Depletion of the high- University of Goias (approval CEP: 895.552). In the abundance plasma proteins was performed through current survey, 100 patients (35M/65F) of both antibody-based affinity columns. Peptide analyses genders and aged 18-90 (M: 30.2) years old were were conducted using a nanoACQUITY UPLC System enrolled after receiving their informed consent. The coupled to a SYNAPT® G2-Si tandem mass Multiplex ligation-dependent probe amplification spectrometer. Protein quantitation was performed by (MLPA) was performed using the P128 CYP450 label-free and proteins with fold-change ≥ 1.5 were MLPA kit (Version C1; MRC-Holland, Amsterdam, considered with differences in abundance. In addition, The Netherlands) according to the manufacturer’s instructions. This gene is located on chromossome identified proteins were ranked through classification 10q23.33 and has 10 exons, but only 4 (1, 7, 8 and 9) algorithms by data mining (DM) analyses. Results: were analyzed. In exon 1 no alteration was found, in High exposure to Pb was observed in workers when the exon 7 was found 2 deletions in homozygosis in compared with non-occupational Pb exposed group men, exon 8 (1 deletion in homozygosis in woman) (mean BLL: 51 and 0.39 µg/dL respectively). The and the exon 9 (1 deletion in heterozygosis in woman). obtained plasmatic proteomic profile showed both This study provides results for CYP2C9 in a plasminogen (PLMN) and α-II antiplasmin (A2AP) population in Goiânia –GO. Hence, it opens up new proteins presented in different abundances in exposed avenues for further investigations by epidemiologists individuals compared to non-exposed group, although in determining inter individual variation in genetic statistics analyses has confirmed these findings only to susceptibility to various diseases caused due to gene- PLMN (p<0.01). Interestingly, a DM demonstrated environment interaction. Is the first time this that PLMN and A2AP were ranked among the most methodology is used, and we believe it is a great tool important proteins to differentiate both groups. for future studies with other genes. Financial support/Acknowledgments: FAPEG. Conclusions: To the best of our Introduction and objective: Lead (Pb) exposure has been associated to neurological, skeletal, hematopoietic and OM 06 - AN EVIDENCE OF PLASMINOGEN- cardiovascular disorders. However, toxic mechanisms PLASMIN SYSTEM LINKED TO LEAD associated to Pb exposure are still poorly understood. TOXICITY BASED ON A HUMAN PROTEOMIC On the other hand, exposure to knowledge, this study STUDY is the first to evaluate effectively the proteome profile in plasma samples of high occupational Pb exposed 1 2 SOUZA, V.C.O. , BARCELOS, G.R.M. , ROSA, subjects. Furthermore, our results allow us to suggest a J.C.3, MAIONE, C.4; BARBOSA, R. M.4; CASSOLI, 5 5 mechanism of Pb toxicity which is for the first time J.S. , MARTINS-DE-SOUZA, D. , BARBOSA proposed. JÚNIOR, F.1 Financial support/Acknowledgments: CNPq,

CAPES e FAPESP 1. School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, USP, Ribeirão Preto- SP, Brazil; 2. Institute of Health and Society, Federal University of São Paulo, UNIFESP, Santos-SP, Brazil; 3. Ribeirão Preto Medical School, University of São Paulo, USP, Ribeirão Preto-SP, Brazil; 4. Institute of Informatics, Federal University of Goiás, UFG, Goiás- GO, Brazil; 5. Institute of Biology, University of Campinas, UNICAMP, Campinas-SP, Brazil.

IMMUNOTOXICOLOGY ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 151

IM 01 - PEROXISOME PROLIFERATOR- reverted when BV2 cells are treated with exogenous ACTIVATED RECEPTOR γ EXPRESSION IS ANXA1. Conclusions: ANXA1 modulates PPARγ MODULATED BY ANNEXIN A1 IN BV2 CELLS expression in BV2 cells and PPARγ is related with phagocytosis mediated by ANXA1. ROCHA, G.H.O. 1; PANTALEÃO, L.N. 1; Financial support/Acknowledgments: Conselho OLIVEIRA, T.2; FESTUCCIA, W.2; Nacional de Pesquisa e Desenvolvimento (CNPq); REUTELINGSPERGER, C.3; SOLITO, E. 4; Fundação de Amparo à Pesquisa do Estado de São FARSKY, S.H.P.1 Paulo (FAPESP, nº 2014/07328-4).

1 Faculty of Pharmaceutical Sciences, Department of Clinical and Toxicological Analysis, University of São IM 02 - EFFECTS OF THE EXPOSURE OF Paulo; São Paulo - SP, Brazil; 2 Department of HYDROQUINONE ON THE CELLULAR AND Physiology, Biological Sciences Institute, University HUMORAL RESPONSE INDUCED BY of São Paulo, São Paulo - SP, Brazil; 3 Department of IMMUNIZATION AGAINST COMMON COLD Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, Maastricht, The FABRIS, A. L. 1; SILVEIRA, E. L.V. 1; FARSKY, S. Netherlands; 4 William Harvey Research Institute, H. P. 1 Barts and the London School of Medicine and Dentistry, Queen Mary University, London, United 1 Universidade de São Paulo, São Paulo - SP, Brasil. Kingdom Introduction: The common cold is a problem of public Introduction: Peroxisome Proliferator-Activated health in Brazil as it affects a large portion of the Receptor γ (PPARγ) is a nuclear receptor expressed in population, especially children and the elderly. Since immune cells, including those in the central nervous 1999, yearly immunization against the Influenza virus system (CNS). Annexin A1 (ANXA1) is a protein was implemented for this specific population. Studies modulated by glucocorticoids that is also expressed in have shown that the exposure to environmental the CNS. Both proteins are known to act as anti- xenobiotics, such as cigarette smoke, can compromise inflammatory factors, but to which extent these factors the efficacy of active immunization. Our research might interact with one another during inflammation is group has studied the toxic mechanisms of not fully understood. Better comprehension of hydroquinone, an immunotoxic agent which can be inflammation expands upon pharmacological options found in high concentrations in cigarette smoke, and is that might be used on dealing with inflammation- also a toxic metabolite of benzene, another inducing toxic agents. Aims: Investigate whether environmental pollutant which can be found in PPARγ expression and its anti-inflammatory effects cigarette smoke as well. Aims: To evaluate if the can be modulated by ANXA1 in a microglial cell line. exposure of male C57BL/6 mice to hydroquinone Methods: BV2 cells (murine microglia) were alters the immune response to the vaccine against the transfected with plasmids aimed to down-regulate Influenza virus. Methods: Mice were exposed to ANXA1 expression. BV2 cells were treated with hydroquinone daily, for eight weeks. They were ANXA1, GW9662 (PPARγ antagonist) and WRW4 vaccinated in the sixth and eighth week through and Boc2 (formyl peptide receptor 1 and 2 antagonists, intramuscular route against Influenza virus. Animals respectively). PPARγ expression was assessed by were euthanized at day 7 and 35 after the last Western Blotting (WB) and Real Time Polymerase immunization and total and specific IgG against Chain Reaction (RT-PCR). BV2 cells were co-cultured vaccine antigen as well as IgG avidity were determined with apoptotic PC12 cells (murine neuron-like cell in serum; quantification of total and specific IgG line); phagocytosis and expression of CD36 (PPARγ secretory cells against vaccine antigen were modulated protein linked with phagocytosis) were determined in spleen, bone marrow and lymph nodes. assessed by flow cytometry and PCR. Expression and Results: Results showed no significant alterations in phosphorylation of STAT6 (transcription factor that IgG avidity and number of specific IgG secreting cells, might link PPARγ and ANXA1) were investigated by although there was increased IgG titer in hydroquinone WB. Results: Results showed that ANXA1 increases exposed group. Conclusions: While hydroquinone protein and gene expression of PPARγ in BV2 cells, exposure increases IgG production, further studies are while lack of ANXA1 decreases such expression. still required in order to assess whether it might Blocking of PPARγ prevents ANXA1 of exerting its influence the immune response, as there was no pro-phagocytosis effects. CD36 surface expression is alteration of IgG avidity after hydroquinone exposure. not modulated by exogenous ANXA1, but gene Financial support/Acknowledgments: Fundação de expression is modulated in a FPR-dependent manner. Amparo à Pesquisa do Estado de São Paulo (FAPESP, Lack of endogenous ANXA1 prevents STAT6 n° 2016/24134-4) expression and phosphorylation, but such effect is ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 152

IM 03 - RHEUMATOID ARTHRITIS IS Financial support/Acknowledgments: CNPQ; AGGRAVATED BY HYDROQUINONE FAPESP (2014/07328-4). EXPOSURE IN THE SENSITIZATION PHASE OF THE DISEASE IM 04 - IMMUNOTOXICITY EVALUATION OF HELUANY, C. S. 1; KUPA, L. V. K. 1; VIANA, M. N. RATS TREATED DURING 28 DAYS WITH 2; FERNANDES, C. M. 2; SILVEIRA, E. L. V. 1; KETAMINE AND/OR ETHANOL FARSKY, S. H. P. 1 ASSOCIATION

1Faculdade de Ciências Farmacêuticas, Universidade MENDES, P. F. 1; SIMON, K. A. 2; HUEZA, I. M. 1,2. de São Paulo, São Paulo - SP, Brasil; 2 Laboratório de Farmacologia, Instituto Butantan, São Paulo - SP, 1. Faculdade de Medicina Veterinária e Zootecnia da Brasil. Universidade de São Paulo (FMVZ-USP), São Paulo - SP, Brasil; 2. Universidade Federal de São Paulo Introduction: Rheumatoid arthritis (RA) is a chronic (UNIFESP), Diadema - SP, Brasil. autoimmune and inflammatory disease and collagen- induced arthritis (CIA) is accepted as an animal model The abuse of substances with psychoactive properties for mimicking RA. Studies indicate an association of for recreational purposes is a public health concern, smoking with disease outcome in humans, where especially when associated with other licit or illicit smoking is the trigger factor in development or as a drugs: the club drugs, characterizing the concept of worsen agent in pre-existing RA. Hydroquinone (HQ) polydrug abuse. In this context, ketamine, an is a phenolic compound, found in high concentrations anesthetic drug used in veterinary medicine, has been in cigarette (where is the major oxidative component), used as a recreational drug, usually associated with and is a benzene metabolite responsible for ethanol. Studies have shown that ketamine or ethanol hematotoxicity of the organic solvent. Objective: has immunomodulatory activities; however, few Investigate the role of HQ exposure on CIA studies were performed with the association of both development in rats and the involved mechanisms. treatments. Thus, considering a polydrug abuse Materials and Methods: Animals were immunized at scenario, we evaluated the immunotoxic effects of this the tail base with 200 µl of bovine type-II collagen association. Wistar rats were distributed into four emulsified 1:1 with complete Freund's adjuvant. A groups (n=8 animals/group), each receiving one of the booster was administered 7 days later. Rats were following treatments for 28 days: K group (30mg/kg of exposed to saline or vehicle (saline-ethanol 20:1) or ketamine, intraperitoneally); E group (10% of ethanol HQ 25ppm, 1 hour/day, using a nebulization chamber in drinking water); KE group, receiving both during 14 days, which comprehended 1 week before treatments; and Control (Co) group, which received collagen injection and 7 days until booster injection. only the vehicles of these drugs. On day 29, animals On 35th day, animals were euthanized and submitted were euthanized for immune evaluation. The to clinical evaluation (score scale of 0-4: 0 = no experimental conditions elicit significant changes on arthritis; 1-2 = weak arthritis; 3 = medium arthritis; 4 = immune parameters regarding E and KE groups when strong arthritis, with all inflamed digits and paws), compared with Co group evidenced by increased in hematological parameters, histological and bone marrow cellularity (**p<0.01 Co vs E and immunofluorescence (IF) analysis of the synovia, *p<0.05 Co vs KE) and lymphocytes count from white quantification of cytokines levels and citrullinated blood cells analyses (**p<0.01 Co vs E and Co vs proteins. Data were obtained in 4 animals per group. KE), as well as a reduction in neutrophils count Results: Data showed that HQ exposure caused (*p<0.05 Co vs E and Co vs KE). No alterations were elevated scores (3-4 scores) of CIA, increased observed in spleen cellularity or relative weights of neutrophils influx into the synovia and levels of IL-6 spleen and thymus. The enhanced cellularity of the and IL-1β in synovial fluid. Also, HQ exposure caused bone marrow can be due to a compensative effect of pannus formation and hyperplasia of synoviocytes, this hematopoietic tissue in replace the reduction of the altered the collagen fiber arrangement on the synovia, circulating neutrophils. Thus, we suggest that in but did not alter the numbers of circulating cells and attempt to suppress the depletion of these neutrophils, citrullinated peptides. In addition, by the IF analysis, an enhancement in bone marrow cellularity was HQ exposure increased levels of AhR receptor, IL-17, occurred. It is know that ethanol can impair CD90 and Ly6G in the synovia in comparison to saline polymorphonuclear cells, interfering in their function or vehicle. Conclusions: our data show that HQ and integrity. Conversely, the increased lymphocyte exposure in the sensitization phase aggravates the count is not caused by any of these treatments, but as a development of RA, suggesting that HQ may be a consequence of the proportionality of the leukogram determinant component of cigarette on RA count. Future experiments will be performed in order development. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 153 to better understand these immunotoxic effects of ethanol associated or not with ketamine. Financial support/Acknowledgments: This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq, Brazil [grant number 140019/2015-4] which provided financial support for the conduct of this research.

MEDICINES AND COSMETICS TOXICOLOGY ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 155

MD 01 - PSILOCYBIN, AYAHUASCA, AND LSD MD 02 - EVALUATION OF TWO RISK IN THE TREATMENT OF ANXIETY, PREDICTION TOOLS OF HEPATOTOXICITY DEPRESSION AND DRUG ABUSE: A SCOPING IN ACETAMINOPHEN POISONING REVIEW NASCIMENTO, A.L.1; SILVA, C.I.1; PACHECO, MAIA, A. H. A; OLIVEIRA, A. F. 1. L.K.1; DOS SANTOS, C.R.1,2.

1. Universidade Federal de Santa Catarina, 1. Setor de Toxicologia, Divisão de Análises Clínicas, Florianópolis - SC, Brasil. Hospital Universitário Professor Polydoro Ernani de São Thiago/UFSC; 2. Departamento de Patologia, Introduction: Anxiety, depression and drug abuse Centro de Ciências da Saúde, Universidade Federal de affect a great part of world population. Treatments in Santa Catarina. this scenario depend on several psychological and pharmacological management that may not achieve Introduction: Hepatotoxicity is a complication of success according to individual susceptibility. Studies acetaminophen toxicity. N-Acetylcysteine (NAC) is with psychedelics have proposed they may be useful used as an antidote, considering the Rhumack- therapeutic tools for treatment of these disorders. Matthew Nomogram risk classification (NRM). Objective: To elaborate a review of studies that However, there are limitations to its application. It has explore the therapeutic potential of psilocybin, been internationally proposed, as an alternative, the ayahuasca, and LSD (lysergic acid diethylamide) by use of the multiplication product of serum conducting psychedelic psychotherapy in patients concentrations of acetaminophen and alanine diagnosed with anxiety disorders, depression, and drug aminotransferase (ALT) dosed at the same time abuse. Materials and Methods: A scoping review was (APAPxALT) as a possible risk predictor. Objective: conducted using proper keywords to search the studies. Evaluate the risk classification of hepatotoxicity in PubMed, SciELO and SpringerLink databases were patients suspected of acetaminophen poisonings, consulted in February 2017. Clinical and observational according NRM and APAPxALT classifications. studies published since 1988 that comprised the Material and methods: Retrospective analysis of the purpose of this review was included after application serum levels of acetaminophen by spectrophotometry, of inclusion and exclusion criteria. Results: From 1013 from September 2014 to June 2017, at the Toxicology titles, ten articles published between 2006 and 2016 Laboratory of the Clinical Analysis Division at Federal were extracted for this study. Participants of the University of Santa Catarina. According to NRM, were studies were oriented on the research and prepared included 35 patients classified with possible and high with psychotherapy. Posteriorly, the evaluated risk of hepatotoxicity. All patients received the NAC. compound was administered at one or more sessions For these patients APAPxALT was calculated. Results: monitored by a research member or during a There were 27 women and 8 men in the group of ceremony, followed by more psychotherapy sessions. patients. The average age was 25 ± 11 years. Follow-up ranged from 24 hours to 12 months after Evaluating the APAPxALT, the following distribution dosing. No serious adverse events were reported was observed: 14 cases (40%) as low risk (10,000 neither intervention was required during any of the mg.L-1 ). Evaluation was not possible to be done in 3 sessions. Validated questionnaires were applied for cases. It was registered 1 case (3%) of death (11,124 evaluation of disorders symptoms, disorders severity mg.L-1) and the other patients were discharged. The or frequency of drug abuse. All studies have shown a average length of hospitalization was 2.5 ± 2.2 days to reduction of the evaluated parameters right after the the ones classified as low risk, 2.6 ± 2.3 days to the administration of the drug with the maintenance of the ones considered possible risk and 4 ± 3,0 to the high results for different periods of follow up according to risk ones. Conclusion: It is possible to observe the each study. Conclusions: Psychedelics seem to present relationship between the APAPxALT values and the potential as an alternative therapeutic tool for disorders time of hospitalization. Even so, 18 patients (51%) in patients without any contraindication and that do not were classified as high risk, hence the indication of respond to conventional treatments, provided that they using the antidote, being possible the improvement of are administered under the supervision of a trained NAC usage. A prospective study should be developed professional. comparing both methodologies, to ensure, in a way, Financial support / Acknowledgments: the use of this new tool in evaluating acetaminophen Departamento de Patologia – UFSC. toxicity. Financial support / Acknowledgments: Centro de Informações Toxicológicas de Santa Catarina e Hospital Universitário Polydoro Ernani de São Thiago, HU-UFSC.

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MD 03 - TOXICOLOGICAL ASSESSMENT OF MD 04 - SYSTEMATIC TOXICOLOGICAL DEGRADATION PRODUCTS IN ANALYSIS FOR IDENTIFICATION OF DESLORATADINE TABLETS: A COMBINED DONATED DRUGS IN BLOOD BANK APPROACH OF COMPUTATIONAL AND EXPERIMENTAL TOOLS REIS B.C.M.1; EFFTING C.2; CUNHA L.C.1.

OLIVEIRA, A. A. F.; ULTRAMARI, M. A. 1. Núcleo de Estudos e Pesquisas Tóxico- Farmacológicas (NEPET), Faculdade de Farmácia, Aché Laboratórios Farmacêuticos, Guarulhos - SP, Universidade Federal de Goiás. 2. Hospital das Brasil Clínicas (HC), Universidade Federal de Goiás.

Introduction: The processes of drug synthesis, Introduction: The consumption of blood and blood manufacturing and storage can undesirably result in products has increased in recent years and it is the formation of impurities and degradation products estimated that in 2010 3,465,172 transfusions were which may impact drug efficacy and safety. performed. According to the PORTARIA Nº 158, of Computational models represent a strong tool to 2016, blood transfusion must be judiciously used due address the toxicity of such compounds considering to the risks to recipients. Systematic Toxicological the limitations of adequate samples to test them Analysis (ATS) has been extensively used for the experimentally and the scarce information in the purpose of identifying and quantifying drugs and other literature, since most of them are still poorly studied. substances in biological fluids. Objective: To estimate Objective: We assessed the toxicity of two degradation the prevalence of drugs in samples of donated plasma products, impurities RRT 2.2 and RRT 3.7, of in blood banks (BS). Materials and Methods: desloratadine tablets to figure out if they represent any Analytical and prospective study, which included 100 risk to patients’ safety. Methods: The impurities were plasma samples, purchased from the HC-UFG BS found above the qualification threshold after 24 (CEP-UFG protocol No. 1,341,581). ATS technique, months in long-term stability study (30ºC, 75% RU). with HPLC-PDA, was used in chromatographic Once the chemical structures were identified, we used conditions validated by Pragst et al., 2001. The mobile an expert rule-based system, Derek Nexus, to carry out phase is composed of potassium phosphate monobasic computational predictions for 59 endpoints in bacteria and acetonitrile. Prior to each analysis, 10 μL of and mammals. We also used a statistic-based system, standard were injected to verify calibration. Three Sarah Nexus, to address specifically mutagenicity per extracts were prepared: acid (hydrochloric acid and ICH M7 guideline. In silico predictions of genotoxicity dichloromethane), basic (Tris buffer and were then checked by Ames test and in vivo dichloromethane) and neutral (acetonitrile), totaling micronucleus assay, following OECD guidelines. 300 samples. After the extractions, 10 μL of each was Results and Conclusions: Two endpoints fired alerts at injected into the HPLC. The detection, differentiation the selected reasoning level. The alert 646 (HERG and identification of the analyte was performed by the channel inhibition) was fired for impurity RRT 2.2, equipment software in the range of 200 to 380 nm whilst the alert 487 (phospholipidosis), was fired for using the UVTOX database, which considers the impurity RRT 3.7. However, in both cases the same relative retention time and UV spectrum of standard alerts were observed in the desloratadine structure, substances in the database. During the analysis of the which means they do not represent additional hazard. results only the compounds with degree of similarity> As to mutagenicity, both impurities were predicted as 0.995 (identical and similar) were considered relevant inactive in bacteria in the two systems, with 44% and for the screening. The data was recorded using the 33% of confidence in Sarah Nexus model for impurity Microsoft Excel® 2007 program and analyzed by RRT 2.2 and 3.7, respectively. This was confirmed SPSS® for Windows®, version 16.0. Partial Results: experimentally in the Ames test performed with the To date, 60 samples of acid extraction were analyzed, product containing both impurities. Additionally, no which identified 35 identical substances (caffeine) and differences were observed in the frequency of micro 68 similar substances; And 75 samples of the nucleated cells in none of the treated groups (500, 800 precipitation extraction (neutral), 6 identical and 1250 mg/kg) in comparison to control. Altogether compounds (caffeine and ethofiline) and 29 similar these data may be useful to support the safety of such compounds. Partial Conclusion: Most samples showed impurities in desloratadine tablets, which are widely some compound, which may interfere in the quality used for the population. and safety of donated blood. Financial support / Acknowledgments: Aché Financial support / Acknowledgments: NEPET, Laboratórios Farmacêuticos HC-UFG Blood Bank and FAPEG

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MD 05 - EFFECTS OF DEET AND ITS MD 06 - EFFECT OF STANDARDIZED FORMULATIONS ON ZEBRAFISH EARLY- EXTRACT OF PROPOLIS (EPP-AF®) ON LIFE STAGE CYTOCHROME P450 (CYP) AND P- GLYCOPROTEIN ACTIVITY IN HEALTHY SOUZA, B. S. 1; BRITO, L. B. 2; OLIVEIRA, G. A. SUBJECTS USING THE COCKTAIL R. 2; MARRETO, R. N. 1; FLEURY, S. R. 1. APPROACH

1. Laboratory of Nanosytems and drug delivery CUSINATO, D. A. C.; ROCHA, A. 1; FILGUEIRA, devices (NanoSYS), Faculdade de Farmácia, G.C.O 1; BERRETTA, A. A 2; LANCHOTE, V.L 1; Universidade Federal de Goiás, Goiânia - GO, Brasil; COELHO, E.B 1. 2. Laboratório de Pesquisa em Métodos Alternativos (LPMALT), Faculdade de Farmácia, Universidade 1 Universidade de São Paulo, Ribeirão Preto, SP; 2 Federal de Goiás, Goiânia - GO, Brasil. APIS FLORA INDL. COML. LTDA.

Topical formulations containing N,N-diethyl-m- Introduction: EPP-AF® is a standardized extract of toluamide (DEET), a mosquito repellent, are related to propolis with established pre-clinical efficacy and neurotoxicity, mainly due its significant absorption safety. Objective: Evaluate the effect of EPP-AF® on through human skin. In addition, the formulation in P-gp and CYP activity, using the cocktail approach. which the active ingredient is dispersed, exerts a Methods: Sixteen healthy adult were investigated remarkably influence in skin permeation, i.e. in DEET before and after exposure to 375 mg/day/p.o of EPP- toxicity. Though, to determine the influence of AF® for 15 days. Blood samples were collected up to formulation in DEET toxicity, zebrafish early-life 12 h after the use of midazolam (0.2 mg), caffeine (10 stage could be used. Thus, the aim of this work was to mg), omeprazole (2 mg), metoprolol (10 mg), losartan evaluate DEET polymer formulations in zebrafish (2 mg) and fexofenadine (10 mg) and LC-MS/MS was model. Formulations containing DEET and polyvinyl used to quantify plasma concentrations of drug probes. caprolactam–polyvinyl acetate–polyethylene glycol Results: Neither matrix effect nor carryover effect (PEG) were prepared by simple dispersion with water. were observed. The methods were linear for the The Fish Embryo Toxicity Test was carried out analytes in the ranges of 0.05 - 20 ng/mL according to OECD TG 236. Groups of 20 embryos (fexofenadine); 0.03 - 5 ng/ml (losartan and E-3174); were exposed in triplicate to negative control (saline 0.1 - 50 ng/mL (omeprazole), 0.3 - 50 ng/mL (5-OH- water) and different concentrations of DEET (0.5 to omeprazole), 0.01 - 10 ng/mL (midazolam), 0.05 - 50 500 µg/mL) dissolved in hydroalcoholic solution ng/mL (metoprolol and α-OH-metoprolol) and 5 - (MeOH 1% v/v) or dispersed in polymer formulation, 1000 ng/mL (caffeine). The pharmacokinetic with PEG concentration varying from 0.65 to 650 parameters of the compounds were calculated using µg/mL. The test was performed in climate chambers at Phoenix® WinNonlin® software. AUC0-t and Cmax 26 ± 1.0°C with a 12:12 hours, light:dark cycle. ratios after and before the EPP-AF® exposure, Parameters such as mortality and developmental presented as geometric mean (CI 90%) were 0.74 (0.62 abnormalities were evaluated for 96 h. No embryotoxic - 0.89) and 0.90 (0.76 - 1.07) for fexofenadine, 0.88 effect was observed in zebrafish exposed to 6.5 and (0.80 - 0.97) and 0.86 (0.76 - 0.98) for losartan, 0.96 650 µg/mL of polymer during 96 h. DEET (0.83 - 1.11) and 0.91 (0.79 - 1.04) for E-3174, 1.18 hydroalcoholic solution showed LC50-96 h of 261 (0.91 - 1.54) and 1.21 (0.87 - 1.70) for omeprazole; µg/mL and no significant developmental abnormalities 1.12 (0.95 – 1.31) and 1.22 (0.95 – 1.67) for 5-OH- were observed in zebrafish early-life stage exposed to omeprazole, 1.14 (1.03 - 1.28) and 1.21 (1.00 - 1.46) DEET concentrations ≤ 50 µg/mL. Formulations for midazolam, 1.04 (0.92 - 1.18) and 0.94 (0.80 - containing up to 93 µg/mL of DEET and 65 µg/mL of 1.12) for metoprolol, 1.05 (0.99 - 1.12) and 0.99 (0.88 PEG did not present any lethal or sublethal effects - 1.12) for α-OH-metoprolol, 0.97 (0.77 - 1.21) and within 96 h. It is possible to conclude that the 0.87 (0.69 - 1.11) for caffeine. AUC0-t metabolic formulation containing a non-embryotoxic polymer ratios of E3174/losartan, 5-OH- and DEET at a concentration 3 times lower than LC50 omeprazole/omeprazole and α-OH- induced no toxicity to zebrafish early-life stage in the metoprolol/metoprolol we found to be, respectively, conditions tested. 1.11 (0.98 - 1.25), 0.94 (0.81 - 1.10 ) and 1.01 (0.88 - Financial support / Acknowledgments: CAPES; 1.16). Conclusion: With the exception of CYP2D6, the CNPq; FAPEG. administration of EPP-AF® under the conditions studied shows CYP2C19 and CYP3A4 inhibition and CYP1A2, CYP2C9 and P-gp induction, although their magnitudes are below the limits defined by the regulatory agencies and therefore exhibit no clinical relevance. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 158

Financial support / Acknowledgments: FAPESP; changes provoked by statin. Moreover, AA FINEP; CAPES; FAEPA; APIS FLORA administered to the fathers was able to restore female reproductive damage at adulthood. Financial support / Acknowledgments: São Paulo MD 07 - REPRODUCTIVE OUTCOMES IN RAT Research Foundation (FAPESP) (Grant #013/22495- FEMALE OFFSPRING FROM MALE RATS CO- 1). EXPOSED TO ROSUVASTATIN AND ASCORBIC ACID DURING PREPUBERTY MD 08 - LONG-TERM TOXICITY OF LEITE, G. A. A.¹; FIGUEIREDO, T. M.¹; PACHECO, VARENICLINE IN ADULT RATS: A T. L.¹; GUERRA, M. T.¹; ANSELMO-FRANCI, J. PHYSIOLOGICAL APPROACH A.²; KEMPINAS, W. G.¹ ZACCARELLI-MAGALHÃES, J. 1; MOREIRA, N. 1. Department of Morphology, Institute of 1; SANDINI, T.M. 2; ABREU, G.R. 3; SÁNCHEZ- Biosciences, Botucatu, São Paulo, Brazil; 2. SARMIENTO, A.M. 1; RICCI, E.L. 3; FUKUSHIMA, Department of Morphology, Stomatology and A.R 4; SPINOSA, H.S. 1. Physiology, School of Dentistry, USP – University of São Paulo, Ribeirão Preto, São Paulo, Brazil. 1. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo - SP, Brasil; 2. Introduction: Dyslipidemias are occurring earlier in the Faculdade de Ciências Farmacêuticas, Universidade de population due to increase of obesity and sedentarism. São Paulo, São Paulo - SP, Brasil; 3. Centro de Rosuvastatin decreases serum cholesterol, however Ciências Biológicas e da Saúde, Universidade several studies have associated statin exposure and Presbiteriana Mackenzie, São Paulo - SP, Brasil; 4 male reproduction impairment. Ascorbic acid (AA) Universidade São Judas Tadeu, São Paulo - SP, Brasil. plays a protective role on male reproductive system. There is a lack of information about the reproductive Introduction: Varenicline is a synthetic chemical effects on rat female offspring of rosuvastatin-exposed substance produced from the alkaloid cytisine, used for animals. Objective: To evaluate if juvenile exposure to smoking treatment, which acts as a partial agonist for rosuvastatin may cause reproductive damage on female α4β2 and α3β4 nicotinic cholinergic receptors and as a offspring and if paternal co-exposure with AA may total agonist for α7 receptor. There are studies ameliorate these possible effects. Material and regarding varenicline’s non-smoking related effects, as Methods: Male rats were randomly divided into six in treatment for the dependence on drugs. However, experimental groups (n=10), which received saline there are no studies in the literature evaluating the solution 0.9%, 3 or 10 mg/Kg/day of rosuvastatin, 150 long-term toxicity of varenicline through a mg/day of AA or 3 or 10 mg/Kg/day of rosuvastatin physiological approach. Objective: The aim of this associated with 150 mg/day of AA from post-natal day study was to evaluate the possible toxicity through (PND) 23 until PND 53. On PND 100, males were hematological, biochemical and anatomopathological mated with female rats to obtain the female pups. The parameters of prolonged exposure (30 days) to day of vaginal opening and first estrus were obtained varenicline in rats. Material and Methods: Forty male and females were euthanized on the first estrus after Wistar rats were treated with varenicline or tap water PND 42 and PND 75 to obtain body weight, organ by gavage for thirty days. They were divided in 4 weights and morphological evaluation of reproductive groups: 3 experimental groups that received different organs. Estrous cyclicity was assessed from PND 60 doses of varenicline (0.03, 0.1 or 0.3 mg/kg) and a until PND 75 and one female per litter were used for control group that received water (n=10 sexual behavior and fertility test. Results: The age of animals/group). On the 30th day of treatment, the vaginal opening and first estrus, body weight and blood and different organs were collected for organ weights at puberty were similar among the hematological, biochemical and anatomopathological groups. Female offspring whose fathers were exposed evaluations. Results: The results show decrease in or co-exposed to the higher dose of statin showed alanine aminotransferase (p<0.05), alkaline lower number of corpora lutea. At adulthood, the phosphatase (p<0.01), total protein (p<0.05) and urea group whose fathers were co-exposed to rosuvastatin (p<0.05) levels in rats that received 0.1 mg/kg and a at the dose of 10 mg showed increased ovarian weight. decrease in alkaline phosphatase (p<0.05) and urea The group whose fathers were treated with 3 mg (p<0.05) levels of rats that received 0.3 mg/kg of showed lower uterine luminal epithelium area. Estrous varenicline when compared to the control group, cyclicity, sexual behavior and fertility showed no however the values are within the normal range of the differences among the groups. Conclusion: Paternal species and the reduction in the levels of these exposure to rosuvastatin at prepuberty delays ovarian parameters lacks biological significance. There were development and diminishes uterine luminal no changes in the other evaluations performed. epithelium in female offspring, suggesting epigenetic ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 159

Conclusion: These data together indicate that Conclusions: The present study’s results suggest that prolonged exposure of rats to different doses of these genes may be used as biomarkers in vitro varenicline was not able to alter hematological, nephrotoxicity, requiring further studies for validation. biochemical and anatomopathological parameters. In addition, our results show toxigenomic is an Financial support / Acknowledgments: FAPESP, important tool to develop new drugs. CAPES e CNPq. Financial support / Acknowledgments: Capes, Fapemig, CNPq

MD 09 - IN VITRO STUDY OF POTENTIAL NEPHROTOXICITY BIOMARKERS THROUGH MD 10 - EVALUATION OF GENOTOXICITY GENE EXPRESSION USING CISPLATIN ASSOCIATED WITH HYDROXYUREA THERAPY IN PATIENTS WITH SICKLE CELL SOARES, S. 1; GROSSI, M. F. 1; NUNES, M. S. 1; DISEASE SOUZA, L. C. R. 1; TAGLIATI, C. A. 1. OLIVEIRA, E. A. M. 1; MACHADO, C. S 1; BOY, 1 Toxicology in vitro Laboratory, Department of K. A. 1; TOGNON-RIBEIRO, R. 1; KELMANN, R. Clinical and Toxicological Analysis, School of G. 1; SANTOS, A. P. P. 3; VELLOSO-RODRIGUES, Pharmacy, Federal University of Minas Gerais, Belo C. 1; GERHEIM, P. S. A. S. 2; MENDONÇA, L. M. Horizonte, Brazil. 1.

Introduction: Toxicological tests are required to 1. Institute of Life Science, Federal University of Juiz develop new drugs. However, the lack of predictability de Fora, Governador Valadares-MG, Brazil; 2. of in vivo models has increased the failure rate of drug Institute of Biological Sciences, Federal University of candidates, increasing time and cost for developing a Juiz de Fora, Juiz de Fora-MG, Brazil; 3. Hemominas new medicine. In vitro alternative methods may lead to Foundation, Governador Valadares-MG, Brazil. the development of safer drugs, reduce cost, and can be used in preclinical trials of new molecules. The lack Introduction: Sickle cell disease (SCD) is a of sensitivity of current markers for nephrotoxicity hemoglobinopathy characterized by structural changes makes its effects are discovered later. Therefore, it is in the hemoglobin molecule. Hydroxyurea (HU) is crucial to search for better early biomarkers. Renal used to stimulate fetal hemoglobin production in system receives large blood flow and therefore it is patients with SCD to improve the hematological important target to many drugs. Whereas the molecular parameters and diminish the pain crises. However, and cellular damage precede the histopathological related potential genotoxic and carcinogenic effects, in changes, the study of gene expression induced by animal experiments and in children, have been drugs has becoming important. Once the expression of identified in previous studies. Thus, currently, a set of genes is consistently changed after the insufficient evidence exists on the long-term risks exposure to xenobiotics, these genes can be used as associated with the use of HU. Objective: The study biomarkers. Hence, nephrotoxic drugs have been used objective was to evaluate genotoxicity in the peripheral such as biomarker studies to identify these genes. blood mononuclear cells (PBMCs) of patients with Cisplatin is a chemotherapeutic agent widely used in SCD treated with HU, using a cytokinesis-block medical practice and has in nephrotoxicity its greatest micronucleus cytome (CBMN-Cyt) assay. Material limitation of use. Objective: Identify possible early and methods: The research subjects were patients with biomarkers of nephrotoxicity and mechanisms SCD attended in outpatient care unit of the underlying the toxic action of cisplatin on renal cell Hemominas Foundation in Governador Valadares-MG, line LLC-PK1. Materials and Methods: LLC-PK1 cell Brazil. We evaluated 14 patients (9 males and 5 line were exposed to cisplatin for 48 hours to 8 females, aged 6–59 years) with SCD treated with HU different concentrations and viability were evaluated (SCDHU group), receiving HU orally at 7.5–19.5 with MTT assay. The expression of nephrotoxicity mg/kg/day (a mean of 12.8 mg/kg/day) for 12–82 related genes was analyzed by quantitative PCR in months (a mean of 44 months). The control group cells exposed at concentrations of 1 uM, 6 uM and 15 comprised 20 patients (12 males and eight females) uM. Results: Decrease in cell viability observed at aged 4–52 years with SCD who did not receive HU concentrations of 1 uM and 6 uM and was not treatment (SCD control group). The blood samples accompanied by changes in gene expression studies were submitted for PBMC isolation, for culture and and require further research for understanding such as evaluation by CBMN-Cyt assay and the determination cisplatin affect cell growth. The HAVCR1 genes of chromosomal damage (micronuclei [MN], (KIM-1), BAX, CASP9, CASP3 and ICAM-1 have nucleoplasmic bridges [NPBs] and nuclear buds increased expression after exposure to 15 mM of [Nbuds ]) in the binucleated cells. Results: The cisplatin when compared to unexposed cells. CBMN-Cyt assay findings for groups SCDHU and ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 160

SCD control for MN, NPBs, and Nbuds, respectively, accessing medications by storing them in places were 10.7±8.4 versus 10.9±8.1; 1.1±0.6 versus beyond their visual range would prevent accidents 1.0±0.8; and 0.3±1.4 versus 1.0±0.9. Thus, the from happening so often. Conclusion: It is evident that frequency of markers of chromosomal damage in the young adults are the most affected by drug groups was similar and the difference was not found to intoxications in the State of Goiás, followed by be statistically significant (p>0.050). Conclusion: accidental intoxication in children. In addition, it is Evidence of genotoxicity in patients receiving clear how much it is valid to invest in measures that hydroxyurea treatment was not found in this study. prevent recurrence, as well as to ensure that more There is less need for major concern about the serious serious outcomes do not occur. long-term consequences of treating patients with SCD Financial support / Acknowledgments: Centro with hydroxyurea, based on the current findings and Universitário de Anápolis the most recent literature. Financial support / Acknowledgments: FAPEMIG (Processes APQ-02608-14, APQ-03560-13), PROPP- MD 12 - AMYLIN: BIODISTRIBUTION, UFJF and Hemominas Foundation. ACTION IN GLYCEMIA AND TOXIC EFFECTS

JOTHA-MATTOS, L; LIMA, L.M. MD 11 - EPIDEMIOLOGICAL PROFILE OS DRUG INTOXICATIONS REPORTED IN THE Federal University of Rio de Janeiro. STATE OF GOIÁS BETWEEN 2011 AND 2015 Amylin is a hormone co-secreted with insulin and as SILVA, L. O.; LIMA, G. C. 1; MARTINS, I. L. O. 1; such has a pharmacological analogue, pramlitide, for OLIVEIRA, R. R. 1; VASCONCELOS, F. G. 1. the treatment of diabetes and other conditions. However, the toxic action of amylin - and its analogue 1 Centro Universitário de Anápolis of commercial use - are still not well described. The objective of this work was to determine the Introduction: The term toxicology is defined as the biodistribution of amylin, acting in glycemia and its study concerning the understanding of toxicants, as possible toxic effects. In order to evaluate the well as their occurrence, behaviors and mechanisms of biodistribution of the exogenous amylin, mice were action. Poisoning consists of a pathological process injected intraperitoneally (ip) with amylin labeled with triggered by endogenous or exogenous substances, iodine 125. The animals were submitted to euthanasia which may even come from commonly prescribed up to 120 minutes after administration. Blood and substances. Objective: Toanalyze data on drug tissues were collected, and radioactivity was measured intoxications in the State of Goiás. Methods: by liquid scintigraphy. To evaluate the action of Descriptive, quantitative, retrospective study of data amylin under glycemia, mice were injected ip with from the Department of Informatics of SUS. Among vehicle or another with amylin. The glycemia was the variables studied are age, municipality of registry determined by glycosimeter using whole blood from and circumstance. Results: Goiânia presented the the tail tip of fasting mouse. To analyze toxicological highest number of occurrences, with 22.51% of parameters, mice were divided into two groups: control records. Then there are Anápolis (14.04%) and Jataí and amylin. The administrations occurred on (8.39%). Regarding the profile of drug intoxications, alternative days during one week, via ip. On the sixth the majority is female (69.97%). Still on the profiles day the animals were submitted to behavioral tests in under analysis, it is noticed that the most affected age Open Field. On the seventh day they were euthanized group comprised individuals between 20 to 39 years and blood and pancreas were collected. All animal (39.03%), followed by children with less than 1 to 9 experiments had protocols approved by the Faculty of years (25.63%). The suicide attempt reaches 56% of Pharmacy Ethics Committee on Animal Use. The notifications. The following are the accidents with biodistribution of amylin decreased with the time and medicines (21.81%), and children have a higher its half-life was determined in 13.58 minutes. From percent age of records in this circumstance (85.46%). organs analyzed, the ones that accumulated most of the The age range of 20 to 39 years is part of a group with molecule were fats (white and brown), muscle, liver high rates of prevalence of depression, a predisposing and pancreas. The pharmacological action over factor to the act of suicide. Reducing the toxicity of glycemia was started from 30 minutes after some drugs, protection mechanisms in drug bottles and administration until 120 (last sampling time). In the marketing control for drugs with toxicity would be toxicological test, animals injected with amylin effective in preventing such events. Accidental showed memory deficit and anxious behavior. poisoning is associated with the second most prevalent Postmortem analyzes suggested subclinical pancreatitis age group and the victim of drug poisoning, children in animals injected with amylin because of changes in less than 1 to 9 years of age. Preventing children from serum lipase and fibrosis in the pancreas. In ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 161 conclusion, the exogenous amylin had its half-life MD 14 - CHARACTERIZATION AND determined and confirmed pharmacological action. COMPOSITION OF GREEN CLAY FOR However, it has also generated toxic effects: COSMETIC USE AND ITS EFFECTS ON behavioral and memory changes and subclinical EMBRYOS AND POST-LARVES OF pancreatitis in mice. ZEBRAFISH (DANIO RERIO) Financial support / Acknowledgments: CAPES MACHADO, M. R. 1; BALDUINO, A. P. Z. 1; RODRIGO PRADO, R. P. 1; MACHADO, M. R. F. 1; MD 13 - STRENGTH TRAINING EFFECT PETRUCELLI, G.C. 1. ASSOCIATED WITH NANDROLONE DECANOATE AND LEPIDIUM MEYENII IN 1. Universidade Federal de Goiás, Regional Jataí BEHAVIORAL HAEMATOLOGICAL PARAMETERS IN WISTAR RATS Introduction: Clays have in their composition metals that provide some sought after aesthetics, standing out MASCARENHAS, M.; FERRÃO, S.K.; in the cosmetic industry. The cosmetic clays used in RODRIGUES, J.L.; BLEMBEEL, A.S.; facial masks must follow some important requirements RUSSOWSKY, A.; RODRIGUES, I.A.; CARDOSO, such as chemical and microbiological safety, V.V. granulometric control and controlled content of heavy metals. It must be in agreement with the National Programa de Pós Graduação em Biociências e Agency of Sanitary Surveillance (ANVISA), which is Reabilitação. Centro Universitário Metodista-IPA the body responsible for the control and inspection of cosmetic products, so they establish a negative list of The spread of strength training and the search for the prohibited substances. Objective: To analyze which perfect body increases the demand for anabolic elements are present in a sample of commercially resources. The use of anabolic androgenic steroids available cosmetic green clay, and to evaluate its occurs often in combination with other substances, chemical and structural composition. And still observe which may increase the side effects. This study aims to its toxicity using embryos and zebrafish larvae. evaluate behavioral parameters in 60 Wistar rats under Materials and methods: chemical and structural treatment with nandrolone decanoate (18mg/kg/week- composition was performed using conventional high- 1, IM), Lepidium meyenni (450mg/kg/week-1, orally) resolution X-ray diffraction (XRD) techniques, Fourier and strength training was realized with 80% of 1MR. transform infrared spectroscopy (FTIR), and X-ray Sixty animals were divided into five groups: sedentary fluorescence with dispersive energy (EDXRF) with the control (SC), strength training (ST), nandrolone use of the fundamental parameters method and the test decanoate (ND), Lepidium meyenii (LM) and strength of release of metals by acid treatment to verify the training, nandrolone decanoate and Lepidium meyenii metals that become bioavailable. To evaluate the (STNL). After 5 weeks of trial, analysis of behavioral toxicity, lethal concentration tests 50 (LC50) were tests (Elevated plus maze, Resident-intruder and object performed using the zebrafish as the experimental recognition) were realized. This tests demonstrated model. Results: The XRD analysis did not indicate the changes in anxiety and stress increase in ST (2.75 ± presence of characteristic peaks of mineral impurities. 0.67) and STNL (2.33 ± 0.33) groups; decreased According to the FTIR analysis, the characteristic band aggressiveness in LM (6.01 ± 1.67) and memory of carbonate and sulfate impurities was confirmed. prejudice in ND (- 0.35 ± 0.16), LM (-0.23 ± 0.16) and And for the analysis of EDXRF was found zirconium STNL(- 0.23 ± 0.21) group. The STNL group STNL metal, listed by ANVISA as prohibited. In showed a significant increase in the number of toxicological tests, pure green clay (50 to 0.025 mg / segmented neutrophils (48,00 ± 4,24) and eosinophils mL) showed an increase in the mortality rate of (4,00 ± 0,01) and decrease in the lymphocytes (32,50 ± zebrafish embryos and post-larvae in long periods of 0,70), bringing injury to the immune system. This exposure and at higher doses, being this time-dose- association needs to be further studied, and in the dependent. Conclusion: It is concluded that a greater meantime, based on these results, is not recommended rigor is required in the supervision of these cosmetics, the use of anabolic steroids or associated with alerting the population about the risks that can be Lepidium meyenii. Cause health. Financial support / Acknowledgments: CAPES, Financial support / Acknowledgments: Apoio FAPERGS, IPA. Financeiro: FAPEG. Agradecimentos: ao Programa de Pós-Graduação de Ciências Aplicadas a Saúde, ao Laboratório Labfish, ao Laboratório de Química.

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MD 15 - FOURIER TRANSFORM INFRARED MD 16 - DETERMINATION OF GENOTOXIC SPECTROSCOPY USING ATTENUATED IMPURITIES BY UHPLC-MS/MS AND TOTAL REFLECTANCE (FTIR-ATR) AS A ANALYSIS OF DISSOLUTION PROFILE IN TOOL FOR CUTANEOUS PERMEATION OF IMATINIB MESYLATE TABLETS COSMETIC AND PHARMACEUTICALS PRODUCTS WOLLF, F. C. 1; DILLENBURG, T. L. 1; ANTUNES, M. V. 1; LINDEN, R. 1; VERZA, S. G. 1. SILVA, MS1; FACCHINI, G1; PINHEIRO, ALTA1; PINHEIRO, AS1; EBERLIN, S2; PICON, FC1; 1. Universidade Feevale, Novo Hamburgo - RS, Brasil EBERLIN, S1. Introduction: Imatinib mesylate (IM) is used for the 1. Kosmoscience Group Campinas/SP, Brazil; 2Sante treatment of gastrointestinal stromal tumors and D’or Institute, Sumaré/SP, Brazil chronic myeloid leukemia since 2001. During the synthesis of IM genotoxic impurities (GTIs), that have Introduction: Actives used in the treatment of potential to cause cancer in humans, can be generated dermatoses act on deeper tissues of the skin, being and represent a toxicological risk even at low levels. necessary to permeate the stratum corneum. On the Furthermore, have been reported sub therapeutic other hand, skin permeation is undesirable in some concentrations of drug which be related to the products, such as sunprotectors and repellents, technological process of drug formulation and can be constituting an important parameter indicative of evaluated by dissolution tests. Objectives: The product safety. FTIR technique is based on the objectives of the present study were to determine GTIs principle of IR absorption by the active, resulting in using UHPLC-MS/MS in IM products and compare characteristic vibrational movements in the molecule. dissolution profile of these products. Methodology: In this sense, the use of FTIR is a useful tool in dermal The content of GTIs (N-(2-methyl-5-aminophenyl)-4- safety assessments, where the objective is that the (3-pyridyl)-2-pyrimidine amine (Imp. 1) and N-[4- active does not permeate the stratum corneum (SC), methyl-3-(4-methyl-3-yl-pyrimidin-2-ylamino)- and in evaluations of topical efficacy with the phenyl]-4- chloromethyl benzamide (Imp. 2) was objective of skin permeation. Objective: Evaluate the analyzed by UHPLC-MS/MS using an Acquity BEH skin permeation of topical products through FTIR- C18 (150 x 2,1 mm, 1,7 um) column, maintained at ATR using ex vivo skin fragments from elective 40°C. The mobile phase consisted of ammonium plastic surgery. Methods: Icaridina and caffeine formate 0.063% (phase A) and acetonitrile plus 0.05% spectrum were measured using the IR formic acid (phase B) in gradient elution at a flow rate spectrophotometer with ATR cell and ZnSe crystal. of 0.2 mL/min. The detection was performed using The baseline spectrum of skin fragments was obtained electrospray ionization in positive ion mode. and then formulations containing each active were Dissolution tests were made in compliance with FDA applied to the skin (2 mg/cm2). Spectrum (Food and Drug Administration) recommendations for measurements were obtained 15 (t15) and 60 (t60) IM tablets. Results and Discussion: Only one minutes after application. In t60, 50 tape-stripping was formulation (D1) failed to meet the requirements and performed for removal SC and new spectral reading. showed 77.22 % of drug dissolved in 30 minutes. Results: The spectrum obtained demonstrate the According to FDA and International Conferences on presence of active on the surface of skin in t15 and t60. Harmonization (ICH) guidelines, GTIs contents should After the 50 tape-stripping process we observed the be limited to a daily dose of 1.0 to 1.5 µg/day. Overall, absence of monitoring bands for Icaridina, which is an the formulations analyzed in this work showed low active used in repellents, confirming the absence of levels of Imp. 1 (≤132.89 ng/tablet) and Imp. 2 (≤ permeability in SC. For assessments of caffeine, an 26.18 ng/tablet). However, formulation D1 contained active ingredient commonly used in cosmetic products very high levels of Imp. 1 (4.26 µg/tablet). Conclusion: that accelerate the lipolysis process, we observed the In order to ensure quality and safety of medicines the presence of the monitoring bands even after removal of quantitative evaluation of GTIs is very important and SC by tape-stripping technique, confirming its skin need to become common during routine analysis of permeation ability. Conclusion: According to the medicines. Periodic verification testing to demonstrate results obtained, we can conclude that the use of the that the products meets specifications is very important FTIR-ATR method together with the tape-stripping to assure that pharmaceutical products not pose technique using human skin fourier transform infrared significant risk to human health. spectroscopy using attenuated total reflectance (FTIR- Financial support / Acknowledgments: PROPPEX - ATR) as a tool for permeation of cosmetic and Universidade Feevale. pharmaceuticals products.

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MD 17 - SYNTHETIC ADULTERANTS MD 18 - DRUG INTERACTIONS IN HOSPITAL PRESENCE, LABELING AND AVERAGE PATIENTS WEIGHT CONFORMITY OF PHYTOTHERAPIC MEDICINE FURQUIM, F.C.1; OLIVEIRA, H.C.2; OKUYAMA, C.E. 1; DINIZ, S.N.1. BORTOLI, S.; FRANÇÓIA, P. C. O.; SORGATTO, P. O.; PEREIRA, A. V. 1. Universidade Anhanguera de São Paulo, Sao Paulo - SP, Brasil; 2 - Hospital Universitario Julio Muller, Universidade Estadual de Ponta Grossa, Ponta GRossa Cuiabá - MT, Brasil - PR, Brasil. Introduction: Drug interactions are considered a public Phytotherapic medicines are an interesting alternative health problem. It can cause potentially harmful effects for minor disorders treatments. The increasing use of to patients and may result in permanent damage, these products is noted with concern, due the fact that clinical worsening, increased hospital admissions and many people use them mistakenly understanding that time of hospitalization. Most studies of drug they do not represent any harm to health and can be interactions only evaluate potential drug interactions use in an uncontrolled manner or without guidance. In and not considering whether or not if the same have addition, several studies have shown the presence of occurred in the patient. Objective: Analyze the synthetic substances in phytotherapic formulations, incidence of actual drug interactions in patients this demonstrates the need for more intense hospitalized in a local hospital of low complexity, in supervision on the production and commercialization Mato Grosso, MT, Brazil. Methods: It was used of these products. The aim of this work was to MICROMEDEX ® Healthcare Series, which classifies investigate the presence of synthetic drugs, as possible the interactions according to the severity and adulterants in herbal products; to evaluate the label and documentary evidence. After this first step, there was a capsule weight variation of phytotherapic medicines daily pharmacotherapeutic monitoring through and their accordance to current legislation. Twenty pharmaceutical anamnesis, analysis diagnostic, and herbal medicinal products were acquired in regular laboratory tests in order to verify that potential market. Synthetic drugs presence as adulterants, as interactions indeed occurred in hospitalized patients. caffeine, chlordiazepoxide, diazepam, fluoxetine and Results: Were identified 2,937 potential drug sibutramine were analyzed by thin layer interactions and found that 28.42% (n = 759) of these chromatography. The label evaluation accordance interactions actually occurred in 87 patients (29%) followed the Brazilian RDC nº 26/2014 requirements with a mean of 8.72 interactions / patient. Among the and the capsule weight variation procedure as patients who had interactions, 62.07% (n = 54) had preconized in Brazilian pharmacopoeia. All of the more than 60 years. In this age group it was also analyzed labels showed some nonconformity. identified 61.27% of all actual interactions observed in According the current legislation about average the study. Also, 70% of patients who had more than 11 weight, 19 of the 20 analyzed samples were in prescription drugs had at least one drug interaction. disagreement with the legislation. 18 had an average Thus, a positive association was observed between age weight below that mentioned on the label and one (p <0.0001) and number of prescription drugs and the sample did not have a weight description. The search occurrence of drug interactions. Conclusion: This of synthetic adulterants, 7 samples showed study demonstrates the need for health staff pay chromatographic profile similar to some of the attention to the fact that drug interactions are investigated synthetic drugs, however in none of them paramount importance for the pharmacological was possible to indubitably confirm the presence of treatment of quality inpatient care, which most often adulterants. This work highlight the need for a more are exposed to polypharmacy, and also shows the need intense inspection on the production and for strengthening of Clinical Pharmacy in Brazil. commercialization of traditional herbal products, since Financial support / Acknowledgments: Anhanguera the label and the average weight of these products Educacional. were in disagreement with the current legislation. In addition, the investigation of possible presence of adulterants in these products is extremely important. Several studies report the use of potentially dangerous drugs as adulterants in phythoterapic medicines, making it a health risk and a public health problem.

MUTAGENESIS AND CARCINOGENESIS

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MC 01 - ASSOCIATION BETWEEN NULL the GSTT1 gene, the same relation was not established GENETIC POLYMORPHISM IN GENES OF (OR=0.742; IC95%=0.492-1.119; p=0.155). XENOBIOTIC METABOLISM (GSTM1 AND Conclusion: This study showed that the presence of GSTT1) AND UTERINE CERVIX CANCER: GSTM1 is a protective factor against uterine cervix META-ANALYSIS cancer. Financial support/Acknowledgments: Fundação de TACCA, A. L. M. 1; SADDI, V. A. 2; VILANOVA- Amparo à Pesquisa em Goiás (FAPEG); Associação de COSTA, C. A. S. T. 3; SILVA, A. M. T. C. 4 Combate ao Câncer em Goiás (ACCG); Hospital Araújo Jorge, Goiânia - GO; Programa de Mestrado 1. Mestre em Ciências Ambientais e Saúde pela em Ciências Ambientais e Saúde (MCAS - PUC/GO). Pontifícia Universidade Católica de Goiás, Goiânia - GO, Brasil; 2. Doutora em Fisiologia, Programa de Mestrado em Ciências Ambientais e Saúde, Pontifícia MC 02 - EVALUATION OF ANTISOLAR AND Universidade Católica de Goiás, Goiânia - GO, Brasil; DNA PHOTO-PROTECTIVE ACTIVITIES OF 3. Doutor em Biologia, Hospital Araújo Jorge, PARTITION FRACTIONS FROM THE Associação de Combate ao Câncer em Goiás, Goiânia ANTARCTIC MOSS Sanionia uncinate - GO, Brasil; 4. Doutor em Biologia Celular e Molecular, Departamento de Medicina, Programa de FERNANDES, A.S.1; MAZZEI, J.L2; FERRAZ, Mestrado em Ciências Ambientais e Saúde, Pontifícia E.R.A3; EVANGELISTA, H.1; FELZENSZWALB, Universidade Católica de Goiás, Goiânia - GO, Brasil. I.1

Introduction: Cancer is a disease of multifactorial 1Laboratory of Environmental Mutagenesis, origin and the significance of genetic and Department of Biophysics and Biometry, University of environmental factors in carcinogenesis is well known. the State of Rio de Janeiro, Rio de Janeiro, RJ; Human papillomavirus (HPV) infection is the main 2Laboratory of Natural Products, Institute of Drug risk factor for uterine cervix cancer. Even so, this virus Technology, Oswaldo Cruz Foundation, Rio de by itself is not enough to lead to cancer. In this Janeiro, RJ; 3Laboratory of Toxicology, Department of context, other factors are considered, such as smoking Pharmacy and Pharmaceutical Administration, and genetic susceptibility. Tobacco and its chemical Pharmacy College, Fluminense Federal University, additivescan be found in the cervical mucus, exposing Niteroi, RJ. the cell’s DNA directly to damage. The enzymes of the Glutathione-S-Transferase (GST) family are Introduction: Moss Sanionia uncinata seems to have responsible for the metabolism of xenobiotics, such as developed protective strategies against UV damage carcinogens present in tobacco, but the genes which through biochemical pathways of secondary encode these enzymes are highly polymorphic and metabolites. This is an important subject for searching have been studied and associated with the risk of natural protective substances that can reduce harmful development of several types of cancer. Objective: To effects of UV exposure as skin cancer. Objective: To evaluate the association of the null polymorphism of investigate the potential of photoprotection, GSTM1 and GSTT1 and the risk of the development cytotoxicity in monolayers and alginate-based three of uterine cervix cancer. Methods: A systematic dimensions (3D) in HaCaT cells, and phototoxicity in literature review and meta-analysis were performed. BALB/c 3T3, by partition fractions (Hexane [Hf], Scientific articles were accessed in the PubMed Dichloromethane [Df], ethyl acetate [Ef], n-butanol database, using the terms "GSTM1"; "GSTT1" and [Bf]) and residual aqueous [Rf] of the hydroalcoholic "cervical cancer". The search covered 10 years of extract of Antarctic moss S. uncinata. Material and publication and 23 articles were selected. From all Methods: For in vitro Sun Protection Factor the these articles, 18 were read in full and, finally, 10 met fractions (1 mg/mL) and/or commercial UV-filters the inclusion criteria to compose the systematic review benzophenone-3 (BP-3, 50 µg/mL), 3-(4- and meta-analysis. Results: The total population methylbenzylidene)-camphor (4MBC, 10 µg/mL), evaluated was 2.247 women, divided into 965 (42.9%) octyl-methoxycinnamate (OMC, 0.01 µL/mL) and cases of cervical cancer and 1.282 (57.1%) healthy octocrylene (OCT, 0.08 µL/mL) were mixed and the women from the control group. The frequencies of the absorbances were measured. Water-soluble tetrazolium null polymorphism in the genes GSTM1 and GSTT1 in salt (WST-1) and lactate dehydrogenase (LDH) assays the group of patients with cervical cancer ranged from were carried out using HaCaT cells. WST dye or 14.3% to 68.8% for GSTM1 and from 15.5% to 59.4% reaction mixture of LDH was added after 24, 48 and for GSTT1. The meta-analysis of the selected studies 72-h exposures of Df, Ef and Bf (0.0004-1 mg/mL) found that the presence of the GSTM1 gene is a and the absorbances were measured. Cell death assays protective factor against cervical cancer (OR=0.757; of HaCaT also were performed with Df, Ef and Bf IC95%=0.628-0.913; p=0.004). On the other hand, for (0.0004-10 mg/mL) in 3D cultures using 1.2% ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 166 alginate. Each fraction and its association with UV- Bhas promotion assay at concentrations ≥ 6 µg/mL, filters were performed by 3T3 neutral red uptake test. while VP did not have any in vitro promoter activity. Results: SPF values of Hf, Df, Ef, Bf and Rf were Cytotoxicity, oxidative stress and endothelial cell 4.5±0.4, 16.1±0.2, 8.0±0.4, 18.7± 1.0 and 2.5±0.3, migration were significantly reduced following VP respectively. The association of Ef, Bf or Rf with BP-3 exposure, compared to 3R4F. The in vitro techniques induced positive synergistic effect. The cell viability of employed were able to distinguish responses between Df, Ef and Bf, evaluated by WST-1 and LDH assays, the VP and 3R4F. The data generated suggests that was above 70% until 0.04 and 1 mg/mL for such products may provide a safer alternative to monolayers or 3D cell culture, respectively. Df, Ef and traditional cigarettes. Further studies investigating the Bf demonstrated photo irritation factor values < 2 (no longer terms effects of VP on consumers is required to phototoxicity), including in association with UV- substantiate disease relevant risk reduction. filters. Conclusion: Df, Ef and Bf are good candidates for searching substances for cosmetic applications mainly in association with UV filters. MC 04 - THE IN VIVO MICRONUCLEUS TEST Financial support/Acknowledgments: FAPERJ. IN MOUSE BONE MARROW AND CHALCONE (E)-3-(3,4-DIMETOXIFENIL)-1-FENIL-2- PROPEN-1-ONA (DMF): MUTAGENICITY AND MC 03 - THE PRE-CLINICAL ASSESSMENT OF ANTIMUTAGENICITY EVALUATION VypeTM ePen, A PRODUCT FOR VAPING NUNES, E.B.1; NODA-PÉREZ, C. 1; CHEN L. C.1; BREHENY, D. 1; GACA, M. 1; THORNE, D. 1; NUNES, W.B. 1; CARDOZO, S.P.1; BERNARDES, MARGHAM, J. 1; PROCTOR, C. 1; SARAIVA, A. 1. A.1; SILVEIRA-LACERDA, E.P.1

1. British American Tobacco, R&D Centre, 1 Universidade Federal de Goiás, Goiânia – GO, Southampton SO15 8TL, United Kingdom. Brasil.

Introduction: VypeTM ePen is a vaping product (VP) Introduction: Chalcones are compounds that have which can be used as a substitute for the experience of attracted great interest in the scientific community due smoking holding a great potential for reducing the to the patchwork of biological activities related. They harm associated with tobacco use but has to be have structures that act on specific targets, such as scientifically proven. Objectives: To characterise the enzymes and chemical mediators, and they are aerosol emissions and assess the biological impact of a interesting candidates for new drugs. Objective: This commercially available VP comparing the results to a paper presents the evaluation of the mutagenicity and reference 3R4F cigarette. Materials & Methods: We antimutagenicity of a chalcone (E)-3-(3,4- assessed over 100 harmful and potential harmful dimetoxifenil)-1-fenil-2-propen-1-ona (DMF) in vivo constituents from the aerosol and compared to 3R4F micronucleus test in the mouse bone marrow. Material aerosol emissions. In vitro toxicological assays were and Methods: For the evaluation of mutagenic activity, used to assess VP´s particulate extracts and total groups of 5 animals were treated, by intraperitoneal aerosols including Ames to determine mutagenic injection, with different doses of chalcone DMF (25, potential; Neutral Red Uptake to measure cytotoxicity, 50 and 100 mg.kg-1 b.w.) and for the evaluation of the the quantification of DNA damage in human lung cells antimutagenic activity were administered the same by assessing phosphorylation of H2AX and the Bhas doses concomitantly with a dose of 4 mg.kg-1 of promotion assay as an in vitro surrogate for detecting mitomycin C, both in 24 and 48 hours. After these tumour promoters. Oxidative stress in lung epithelial times the bone marrow are collected, prepared, stained cells and endothelial cell migration were assessed as and analyzed After these times the bone marrow are key events related to DPOC and cardiovascular collected, prepared, stained and analyzed by optical disease, respectively. Results & Conclusions: microscopy. The genotoxicity was evaluated by the Chemical analysis demonstrated substantial reduction frequency of micronucleated polychromatic in constituents in the Vype ePen aerosol relative to erythrocytes (EPCMN) and cytotoxicity was analyzed 3R4F smoke. Furthermore, data from in vitro assays by the polychromatic erythrocytes and indicated that the biological response to VP aerosol normochromatic ratio (EPC / ENC). This study as was significantly lower relative to 3R4F smoke. VP approved by the animal experimentation review board was non-mutagenic in the Ames assay using strains of our institution. Results and Conclusion: The results TA98 and TA 100 with metabolic activation (S9), of the assessment of the mutagenicity showed that all however 3R4F was mutagenic. Increasing doses tested of chalcone DMF, significantly increased concentrations of 3R4F smoke demonstrated a dose the number of EPCMN (p< 0.05) compared with the response in DNA damage-induction, in contrast, VP negative controls. Concerning the antimutagenicity did not induce DNA damage. 3R4F was positive in the capacity/ effect, the doses of 25 and 50 mg.kg-1 b.w. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 167 after 24 hours significantly reduced the frequency of 0.45 µm membranes filtration. The test was performed EPCMN, for the positive control group (p 0.05), while in dose-response experiments according to ISO the dose of 100 mg.kg-1 b.w. co-administered MMC 16240:2005, in absence and presence of metabolic enhanced the mutagenic action. The cytotoxicity was activation system (-/+ S9). The active compounds observed only for dose 50 mg.kg-1 b.w. co- N,N-dimethyltryptamine, harmaline, harmine, and administered MMC in 48 hours. The potential of DMF tetrahydroharmine were quantified, by HPLC-DAD, in in genetic material are still poorly understood. all the beverage samples. Both ayahuasca samples Therefore, other studies should be performed to define were mutagenic for the strains TA98 and TA100 in the their mechanisms of action and properties. presence and absence of S9 and for TA102 only +S9. Financial support/Acknowledgments: CNPq. The beverage prepared from B. caapi had the same ayahuasca mutagenicity profile, and the beverage prepared from P. viridis had no mutagenic activity. MC 05 - MUTAGENICITY OF AYAHUASCA Our results demonstrate that ayahuasca has mutagenic BEVERAGE TO THE SALMONELLA activity and that this activity is related to B. caapi. The MICROSOME ASSAY USING THE OECD negative results obtained with line YG1041 indicate RECOMMENDED STRAINS that the mutagens present in the samples do not belong to the class of nitro-compounds and aromatic amines. KUMMROW F 1; MASELL; BS 2; LANARO R 3; Our results also indicate the need for a detailed COSTA JL 4; UMBUZEIRO GA 5; A LINARDI A 6. toxicological evaluation of this beverage. Financial support/Acknowledgments: The authors 1Institute of Environmental, Chemical and acknowledge FAP of Santa Casa de São Paulo School Pharmaceutical Sciences, Federal University of São of Medical Sciences by the financial support. Bianca S. Paulo (Unifesp), Diadema-SP, Brazil. 2Pharmaceutical Maselli acknowledges CAPES by the PhD scholarship. Sciences Faculty, University of São Paulo (USP), São Paulo-SP, Brazil. 3Poison Control Center, Faculty of Medical Sciences, State University of Campinas MC 06 - IN VITRO INVESTIGATION OF THE (Unicamp), Campinas-SP, Brazil. 4Faculty of CYTOTOXIC POTENTIAL OF THE RED Pharmaceutical Sciences, State University of PROPOLIS OF THE STATE OF PERNAMBUCO Campinas (Unicamp), Campinas-SP, Brazil. 5School of Technology, State University of Campinas BARROS, J. V. 1; ARAÚJO, J. R. S. 1; (Unicamp), Limeira-SP, Brazil. 6Department of BRASILEIRO-VIDAL, A. C. 1; ARAÚJO S. S. 2; Physiological Sciences, Santa Casa de São Paulo SANTOS, N. 1. School of Medical Sciences, São Paulo-SP, Brazil. Ayahuasca is a beverage traditionally used in the 1. Universidade Federal de Pernambuco, Recife - PE, religious ceremonies of some Amazonian peoples. Brasil; 2. Universidade Federal Rural de Pernambuco, However, its use has recently spread around the world. Recife - PE, Brasil. The ayahuasca is obtained from decoctions of the Banisteriopsis caapi that contains β-carbolines Propolis is a substance composed of resin, balsam, (harmine, harmaline and tetrahydroharmine) plus wax, essential and aromatic oils, pollen and impurities Psychotria viridis that contains N,N- being harvested by honey bees from different exudates dimethyltryptamine, a potent hallucinogen. The of plants and used as protection for the hive, to seal the therapeutic use of ayahuasca has been proposed, walls and embalm Dead invaders. Propolis is widely however the safety of its use must be proven. The marketed as an alternative medicine worldwide and is genotoxicity evaluation is mandatory to assess its available in the form of capsules, extract, oral safety. The Salmonella/microsome assay is antiseptic, throat lozenges, creams and powders. Thus, recommended by the Organisation for Economic Co- the therapeutic effect of propolis on some diseases operation and Development (OECD - Guide 471), both claimed by folk medicine has increased interest in for evaluation of pure compounds and complex understanding the biological properties of this mixtures. Our objective was to evaluate the substance making it necessary to investigate its ability mutagenicity of two independent ayahuasca samples, to cause possible changes in cell structure and viability and also of beverages prepared with each of the plants after exposure. Therefore, this study aimed to evaluate separately. For this purpose, the five Salmonella the cytotoxic potential of Pernambuco's red propolis strains recommended by the OECD (TA97a, TA98, through the MTT test ([3-(4,5-dimethylthiazol-2yl)- TA100, TA102 and TA1535) plus the YG1041, which 2,5-diphenyl tetrazolium bromide]). Peripheral selectively detect the mutagenicity of nitro-compounds mononuclear cells were isolated from twelve mL and aromatic amines, were used. All the beverage human blood, which were seeded in 96-well samples were prepared as they are used in religious microplates and treated with varying concentrations of ceremonies and before the tests were sterilized using the red propolis extract (0.39, 0.78, 1.56, 3.12, 6.25, ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 168

12.5, 20, 30, 40, 50, 100, 150, 200, 250 and 300 positive control were used RPMI 1640 and Fetal μg/mL). As negative, positive and solvent control, Bovine Serum (4:1; v:v) and 1% Triton X-100, RPMI 1640 medium, 1% Triton X-100 and 1% DMSO respectively. As solvent control, methanol was used at (dimethylsulfoxide), respectively, were used. After 24 2%. While the mutagenicity was evaluated by the h of exposure to the treatments, the cells were micronucleus (MN) test at 400, 200 and 100 μg/mL of incubated with 20 μl of MTT at 5 mg/mL for 3 h, after the EtOAc fraction. RPMI 1640 and Fetal Bovine which 100 μL of DMSO was added to each well. Serum (4:1; v:v) were used as negative control and Subsequently, the plate was subjected to the methyl methanesulfonate (4x10-4) were used as spectrophotometer reader using the 570 nm filter. The positive control. In relation to cytotoxicity, the EtOAc absorbance values of each concentration tested were and MeOH fractions showed cytotoxicity for compared to the negative control and showed that the concentrations over 1000 μg/mL and 400 μg/mL, red propolis extract was only considered cytotoxic respectively. The micronucleus test revealed that none from the concentration of 150 μg/mL, which presented of the EtOAc fraction concentrations induced 72% cell viability. The performance of genotoxic and significant changes comparing to the negative control. mutagenic studies is necessary as a way to complement Thus, the observed results suggest the safe doses of A. the MTT test and establish a concentration of use that cearensis EtOAc and MeOH fractions for use in does not cause damage to human health. It was also therapeutic processes and in the formulation of future observed a possible protective effect in the medicinal products. concentrations of 12.5, 20, 30 and 40 μg/mL, since Financial support/Acknowledgments: CNPq. cellular viability was superior to the negative control, but antigenotoxicity studies are necessary to confirm this hypothesis. MC 08 - MINIATURIZATION OF THE Financial support/Acknowledgments: CNPq. MICROSUSPENSION SALMONELLA / MICROSOME ASSAY IN AGAR MICROPLATES – MPA MC 07 - CYTOTOXICITY AND GENOTOXICITY EVALUATION OF LEAF ZWARG, J.R.R.M.; MORALES, D. A. 1; ETHANOLIC EXTRACT FRACTIONS FROM UMBUZEIRO, G. A. 1. AMBURANA CEARENSIS (ALLEMAO) A.C.SM. 1. School of Technology, UNICAMP, Limeira – SP, ARAUJO, J. R. S. 1; BARROS, J. V. 1; ALMEIDA P. Brazil. M. 2; SANTOS, N. 1; LIMA, C. S. A. 1; BRASILEIRO-VIDAL A. C. 1. Amburana cearensis (Allemao) A.C.Sm. (Fabaceae) is widely used in folk medicine for asthma, cough and 1. Universidade Federal de Pernambuco, Recife - PE, bronchitis treatment. In addition, its leaves have Brasil; 2. Universidade Estadual do Piauí, Teresina - antioxidant and antimicrobial properties. However, PI, Brasil. studies on cell viability and damage to genetic material have not yet been performed. Thus, the aim of the Amburana cearensis (Allemao) A.C.Sm. (Fabaceae) is present study was evaluated the cytotoxic and widely used in folk medicine for asthma, cough and genotoxic effect of the fractions obtained from A. bronchitis treatment. In addition, its leaves have cearensis leaf crude ethanolic extract on peripheral antioxidant and antimicrobial properties. However, blood mononuclear cells from human blood. Leaves studies on cell viability and damage to genetic material were extracted in ethanol (96%) and concentrated in a have not yet been performed. Thus, the aim of the rotary vacuum evaporator (≤ 45ºC) for crude ethanolic present study was evaluated the cytotoxic and extract (BSE) production. The hexane (Hex), genotoxic effect of the fractions obtained from A. dichloromethane (Dec), ethyl acetate (EtOAc) and cearensis leaf crude ethanolic extract on peripheral methanol (MeOH) fractions were obtained from the blood mononuclear cells from human blood. Leaves solubilization of BSE every 12 h in respective solvents were extracted in ethanol (96%) and concentrated in a and concentrated in a rotary vacuum evaporator. rotary vacuum evaporator (≤ 45ºC) for crude ethanolic Cytotoxicity was evaluated by the MTT assay (5x105 extract (BSE) production. The hexane (Hex), cells/well) at 50 to 2000 μg/mL concentrations of the dichloromethane (Dec), ethyl acetate (EtOAc) and EtOAc and MeOH fractions. For the negative and methanol (MeOH) fractions were obtained from the positive control were used RPMI 1640 and Fetal solubilization of BSE every 12 h in respective solvents Bovine Serum (4:1; v:v) and 1% Triton X-100, and concentrated in a rotary vacuum evaporator. respectively. As solvent control, methanol was used at Cytotoxicity was evaluated by the MTT assay (5x105 2%. While the mutagenicity was evaluated by the cells/well) at 50 to 2000 μg/mL concentrations of the micronucleus (MN) test at 400, 200 and 100 μg/mL of EtOAc and MeOH fractions. For the negative and the EtOAc fraction. RPMI 1640 and Fetal Bovine ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 169

Serum (4:1; v:v) were used as negative control and MC 10 - PATTERN OF EXPRESSION AND methyl methanesulfonate (4x10-4) were used as METHYLATION IN MALE BREAST CANCER positive control. In relation to cytotoxicity, the EtOAc and MeOH fractions showed cytotoxicity for SILVA,L.A. 1; PASSOS,X.S. 1; concentrations over 1000 μg/mL and 400 μg/mL, CORDEIRO,A.M.T.S. 1 respectively. The micronucleus test revealed that none of the EtOAc fraction concentrations induced 1. Universidade Paulista de Goiânia. significant changes comparing to the negative control. Thus, the observed results suggest the safe doses of A. Male breast cancer is a rare disease that accounts for cearensis EtOAc and MeOH fractions for use in 1% of all breast tumors. Breast neoplasm in men is therapeutic processes and in the formulation of future related to klinefelter syndrome, obesity, gynecomastia medicinal products. and the prostate tumor. This work aims to understand Financial support/Acknowledgments: The authors the specific expression and methylation patterns in thank FAPESP Project 2013/16956-6. José Ricardo R. male breast cancer; Confirm the influence of the M. Zwarg thanks FAPESP Project 2015/11399-7 for methylase on the PIK3CA gene and characterize the the IC scholarship. Daniel A. Morales thanks CAPES most frequent mutations associated with this neoplasm. for the PhD scholarship. The SOLUTIONS project has This study is a narrative review that includes articles received funding from the European Unions Seventh published in PubMed and oncology journals from 2012 Framework Programme for research, technological to 2017. The results demonstrate that 97% of the development and demonstration under grant agreement lesions are ductal carcinoma in situ. Of the neoplasms no. 603437. studied, 80% had a function gain mutation in BRCA2. Only 10% of the tumors evaluated are HER2 positive. Tumors expressing Ki67 had a worse prognosis. MC 09 - METILATION STANDARD GENE Patients demonstrated better response to tamoxifen EXPRESSION IN MEN'S BREAST CANCER treatment. Two significantly associated tumor polymorphisms rs13387042 and Rs3803882 may aid in SILVA, L.A. 1; PASSOS, X.S. 1; SILVA, A.M.T.C.1. the understanding of mutagenesis and carcinogenesis of breast neoplasm in men. We conclude that there is 1. Universidade Paulista de Goiânia. no congruence between male and female breast cancer; In situ carcinoma patients classified as luminal 2 who Male breast cancer is rare, about 1% of all cases of expressed HER2 are the patients who presented the breast tumors. This study analyzed the pattern of best prognosis and RASSF1A may be the key to methylation and expression in tumor suppressor genes understanding the regulation of miRNAs. and oncogenes. The studies evaluated hypermethylated genomic regions and aims to understand the methylation of DNA influencing gene expression. It is MC 11 - A STEP CLOSER TO NAMI-A: NOVEL a systematic review that used articles indexed in COMPLEX BASED ON RUTHENIUM PubMed from 2012 to 2017. Analyzing the PYRAZOL-PYRIDINE methylation of gene promoters may help to understand the biology of the tumor and contribute to a REZENDE, R.M.R. 1; LIMA, A.P. 1; TRAVASSOS, standardization of more frequent mutations in the I.O. 1; NORONHA, L.F. 1; DE SOUZA, A.R. 1; already diagnosed tumors. The BRCA2 gene was the SOUZA, A.R. 1; MENEGATTI, R. 1; MOURA, most cited gene and the point mutations at specific R.D.P. 1; CARVALHO, F.S. 1; LIÃO, L.M. 1; sites of the gene are associated with the onset of the SILVEIRA-LACERDA, E.P. 1. tumor. The objective of this study is to improve the prognosis and diagnosis of these patients with the 1 Universidade Federal de Goiás, Goiânia-GO, Brasil. standardization of the most frequent mutations. It is concluded that the method was able to evaluate the NAMI-A was the first ruthenium interference of methylation in intergenic regions in metallopharmaceutical to enter clinical trials, showing hypoexpressed genes and their ability to inhibit the interesting antimetastatic properties. This transcription of oncogenes. antimetastatic activity, along with its relatively mild toxicity in animal models, were very promising results. However, the phase II clinical studies were disappointing as they were classified so insufficient, since the efficacy of the treatment was lower than expected. From these results, it was necessary to take a step forward to design new complexes that incorporate the positive characteristics of NAMI-A, so the present ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 170 study aimed to evaluate a new complex of Ruthenium, male rats randomly assigned to the study, divided into analogous to NAMI-A, in order to antitumor screening seven groups, each with 6 animals. The animals were and antimetastatic activity. In the cytotoxicity assay, kept in a controlled temperature room under light- ± 12 the LFN12 complex and NAMI-A were evaluated hours and with free access to adequate water and feed against three tumoral strains: murine ascitic tumor throughout the study period. The animals were treated (Ehrlich), human alveolar lung carcinoma (A549) and daily with Cissus quadrangularis orally (gavage) for 4 murine melanoma (B16-F10) and a normal line: lung days at doses of 500,1000,1500, 2000 mg / kg and on fibroblast (L929) using the cell viability assay the last day cyclophosphamide at a dose of 50 mg / kg technique (MTT reduction method). Low cytotoxicity was administered intraperitoneally. As euthanasia the was observed for most of the cell models tested, with decapitation method was used, peripheral blood and IC50 (Inhibitory Concentration) values (> 200μM); It bone marrow were collected to be used in mutagenic was not possible to calculate the IS (Selectivity Index), and antimutagenic analyzes with the micronucleus test. since the IC50 of the compounds were high. The Data were analyzed using the Kolmogorov-Smirnov Scratch Test was also performed, which consists of a test, ANOVA and the Tukey test as post hoc. The method to investigate cell migration. The ruthenium results were verified by mean, frequency (%) and complexes against the B16F10 line were tested at standard deviation (SD), using SPSS and considering a concentrations 1, 10 and 100 μM for 0, 24 and 48h. In value of p < 0.05. The results demonstrated were 24h, 100 μM of the LFN12 complex was shown 80.2% significant when compared to the control groups, ± 7.2 of open area and for NAMI-A 83.7% ± 5.2. In 48 demonstrating that as the dose increased to one, the h an open area for LFN12 was presented in the amount of micronuclei increased in both the peripheral concentration of 100 μM 51.2% ± 10.5 and for the blood and the marrow. For decades plants are being NAMI-A 53.9% ± 10.9. This decrease in the cellular used as a medicine and believed to be safe but these expansion found can be correlated with inhibition of plants still require scientific information with further metastases in vivo. Thus, the LFN12 analogue study. analogous to NAMI-A may be a probable drug against Financialsupport/Acknowledgments: IPA; CAPES; the investigated tumor models, with possible anti- FAPERGS. metastatic activity. Financial support/Acknowledgments: CNPq. MC 13 - EVALUATION OF THE CYTOTOXIC EFFECT OF THE AQUEOUS EXTRACT OF MC 12 - MUTAGENIC EVALUATION OF LIBIDIBIA FERREA ON PERIPHERAL EXTRACT OF Cissus quadrangularis (Vitaceae) MONONUCLEAR CELLS (PBMCS) USING THE IN WISTAR RATS SUBMITTED TO MTT TEST SUBCHRONIC TREATMENT COSTA, R. F. 1; ARAUJO, J. R. S. 1; SPINDOLA, D. MASCARENHAS, M.A.; SANTOS, K.; C. 1; BARROS, J. V. 1; MELO, M. S. 1; SILVA, M. RODRIGUES, I.; PANAZOLO, M.; ROXO, L.; V. 1; BRASILEIRO-VIDAL, A. C. 1. JUSTO, J.; BLEMBEEL, A.; GERSON, S.;L GAUDENZ, R.; CARDOSO, V.V. 1. Universidade Federal de Pernambuco, Recife - PE, Brasil. Programa de Pós-Graduação em Biociências e Reabilitação. Centro Universitário Metodista-IPA Libidibia ferrea (Mart. Ex Tul) L. P. Queiroz (Fabaceae), known as pau-ferro-verdadeiro, showing Medicinal plants represent a factor of great importance great medicinal importance, due to the antimicrobial, for the maintenance of the health conditions of the anti-inflammatory and antitumor activities of its people. Besides proving the therapeutic action of secondary metabolites. However, there is lack of several popular plants, phytotherapy represents an information regarding the cytotoxic activity of its important part of a people's culture. Cissus extracts. Thus, the present work aimed the evaluation quadrangularis is a vine of the family Vitaceae, its of the cytotoxic potential of the aqueous extract of the extracts and powders have been used to promote the L. ferrea fruit on mononuclear cells of peripheral treatment of bone and tissue diseases, besides human blood (PBMCs) using MTT [3-(4,5- presenting several actions as analgesic, anabolic and Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium weight loss promotion, its chemical constituents show Bromide] cell viability test. Peripheral blood efficient effects Antioxidants. The objective of this mononuclear cells, separated for density gradient study was to evaluate the mutagenic effect in different (Histopaque-1077®), were added in a 96 well plate (5 concentrations of Cissus quandrangularis extract with x105 cells per well) with different concentrations of subchronic exposure in rats. The experimental study the extract of the L. ferrea fruits: 50, 45, 40, 35, 30, 25, was performed with quantitative data, using 42 Wistar 20, 15, 10, 5, 2.5 and 0.5 mg/mL, diluted in RPMI ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 171

1640: Fetal Bovine Serum (4:1; v:v). For the negative 3.12 and 6.24 mg/mL) of the cyclohexanic fraction of and positive controls, RPMI 1640: Fetal Bovine Serum the A. colubrina’s foliar extract and incubated in a (4:1; v:v) and 1% Triton X-100 were used, humid atmosphere (37ºC and 5% CO2) for 24 h. respectively. They were incubated at 37°C at 5% CO2 DMEM and RPMI culture mediums were used as for 24h. Then, MTT was added to a 5 mg/mL final negative control and Doxorubicin and Triton X-100 concentration and incubated again for 3h. were used as positive control. As solvent control, ethyl Subsequently, the medium was discarded, 100 μL of acetate was used. After exposure, 20 μL of the MTT DMSO were added to the wells and the measurement dye were added to the wells for 3 h and the microplate was done in spectrophotometer (570 nm). was centrifuged. Then, 100 μL of DMSO was also Concentrations up to 20 mg/mL had no cytotoxic added and the microplate was subjected to effect on PBMC cells, since cell viability remained spectrophotometer reading using a 570 nm filter. The above 80% (ranging from 91.2 to 100%). On the other absorbance values of each well were compared with hand, concentrations higher than 20 mg/mL reduced the negative controls. The cyclohexanic foliar extract the viability in an inverse proportion to the extract of A. colubrina had a cytotoxic effect in HL-60 cells at concentration, ranging from 75.8% for 25 mg/mL to 3.12 and 6.24 mg/mL, with cell viability of 63% and 57.8% for 50 mg/mL. Therefore, caution is 15%, respectively. On the other hand, macrophages recommended for medicinal purpose, since high and PBMCs showed no cytotoxicity when they were dosages of L. ferrea fruits could display cytotoxic exposed to the different concentration tested. Thus, it effects. In addition, additional tests are required to is concluded that the cyclohexane extract has check for genotoxic effects of the dosage tested. antitumor potential. Additional studies with Financial support/Acknowledgments: CNPq. micronucleus tests using PBMC are being performed to verify possible genotoxic effect of the extract on normal human cells. MC 14 - EVALUATION OF THE CYTOTOXIC Financial support/Acknowledgments: CNPq. EFFECT OF THE AQUEOUS EXTRACT OF LIBIDIBIA FERREA ON PERIPHERAL MONONUCLEAR CELLS (PBMCS) USING THE MC 15 - RUTHENIUM II PROTOTYPE ACTS MTT TEST ON TUMOR CELLS WITH CELL CYCLE PAUSE IN G0 / G1 AND INDUCES APOPTOSIS WITHOUT DEMONSTRATING CYTOTOXIC CORREIA, D. S. 1; COSTA R. F. 1; ARAÚJO, J. R. OR GENOTOXIC EFFECTS IN HEALTHY S. 1; BARROS, J. V. 1; ARAÚJO S. S. 2; SILVA, C. CELLS M. A. 2; SILVA, M. V. 1; BRASILEIRO-VIDAL, A. C. 1. GOVEIA, R. M. 1; PIRES, W. C. 1; ALMEIDA, V. P. 1; SILVEIRA-LACERDA, E. P. 1 1. Universidade Federal de Pernambuco, Recife - PE, Brasil; 2. Universidade Federal Rural de Pernambuco, 1. Universidade Federal de Goiás, Goiânia – GO, Recife - PE, Brasil. Brasil.

Anadenanthera colubrina (Fabaceae family), also Cancer is the second largest death cause in the world. known as angico, is one of the typical woody species Preclinical studies with ruthenium complexes have of the Caatinga biome that shows recognized shown promising results in the treatment of cancer antimicrobial activity. In popular medicine, the aerial with less toxicity than the usual platinum complexes. parts of A. colubrina are used in the treatment of The objective of this study was to evaluate a new anemia, cancer and inflammations in general. ruthenium II complex, called LGMC-53, observing its However, the possible adverse or beneficial effects that cytotoxic potential by MTT, genotoxic by the alkaline this species can cause to human health are not known. comet method and the mechanism of action through Thus, the present study aimed to evaluate the cytotoxic the cell cycle, annexin-V and gene expression tests potential of different concentrations of A. colubrina’s .The MTT test showed an IC 50 of 14.94 μM for S-180 leaf extract using the cell viability test (MTT) in tumor cells and 16.44 for L-929 normal cells. In the human cells (normal and tumorcultures), in order to cell cycle assay using sarcoma 180 (S-180) thegrowth establish concentrations of safe use with possible was observed in the G0 / G1 phase at concentrations of anticancer effect. For the experiment, tumor cells 14 μM and 28 μM (53% and 72%) higher than the derived from promyelocytic leukemia (HL-60) and control (22%) in 24 hours of exposure. In the control, macrophages (RAW 264.7) as well as peripheral 58% of the cells were in synthesis, at concentrations of mononuclear cells (PBMC) from human blood were 14 and 28 μM, 34% and 14% of cells were obtained at seeded in 96-well microplates. The cell lines were this stage. In 48 hours the same behavior was exposed to different concentrations (0.39, 0.78, 1.56, maintained. In the annexin-V test using s-180, the ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 172 negative control had 94.27% of viable cells, 1.88% in for 28 days. The micronucleus test was performed in initial apoptosis and 1.94% in late apoptosis. There bone marrow and the cells were analyzed by light was a significant difference when compared to LGMC- microscope. This test was approved by the Ethics 53 in the concentration of 20 μM, where 70.91% of Committee in Animal Experimentation from the viable cells, 12.26% of initial apoptosis and 11.44% of Federal University of Grande Dourados (protocol: late apoptosis were observed. The results of qPCR, 29/2015). Results: The negative control groups using S-180, show a significant increase in the presented an average of 2.2 ± 0.37 and 3.0 ± 0.31 of expression of the studied genes (Caspase 3, Caspase 8, micronucleated polychromatic erythrocytes (MNPCEs) Caspase 9 and Tp53), with the exception of Bax at the for female and male rats, respectively. The positive concentration of 14 μM. All genes that experienced control groups, as expected, presented an average of significant increase in expression were expressed late. 11.60 ± 0.81 and 12.0 ± 0.83 of MNPCEs for female Caspase 9 was the gene with the greatest increase in and male rats, respectively. All groups treated with the expression, appearing 4.57 times extended if compared doses of AEAE were statistically similar to the to control despite the fact that Bax had its expression negative control group. The positive control group, decreased by 0.41 times compared to control. With the again as expected, decreased the production of new healthy cell comet assay L929, after 24 hours of erythrocytes. The estimated ratio of PCE/NCE in bone exposure, it was observed that there was no significant marrow preparations showed no statistical changes in genotoxicity at any of the treated concentrations (14 erythropoiesis after the administration of the AEAE. μM, 24 μM). It can be concluded that the LGMC-53 Therefore, groups treated with the AEAE were complex induce to cell death by apoptosis with cell statistically similar to the negative control groups. cycle arrest in G0 / G1 and causing fewer damages Conclusion: Therefore, the AEAE presented low against healthy cells. cytotoxicity, and mutagenicity at the doses tested in Financial support/Acknowledgments: CAPES. rats. Financial support/Acknowledgments: CAPES and CNPq. MC 16 - EVALUATION OF THE MUTAGENICITY OF ALIBERTIA EDULIS EXTRACT IN RATS MC 17 - GENOTOXIC EVALUATION OF ALIBERTIA EDULIS AQUEOUS EXTRACT MENEGATI, S.E.L.T.1; LIMA, F. F. 2; TRAESEL, THROUGH THE COMET ASSAY G. K. 2; ARAÚJO, F. H. S. 3; MELLO, R. M. 4; GASPAROTTO JÚNIOR, A. 2; DALSENTER, P. R. MENEGATI, S.E.L.T.1.; LIMA, F. F. 2; TRAESEL, 1; OESTERREICH, S. A. 2 G. K. 2; ARAÚJO, F. H. S. 3; MELLO, R. M. 4; GASPAROTTO JÚNIOR, A. 2; DALSENTER, P. R. 1 Universidade Federal do Paraná, Curitiba - PR; 2 1; OESTERREICH, S. A. 2; VIEIRA, M. C. 2 Universidade Federal da Grande Dourados, Dourados - MS; 3 Universidade Federal do Mato Grosso do Sul, 1 Universidade Federal do Paraná, Curitiba - PR; 2 Campo Grande - MS; 4 Universidade Tecnológica Universidade Federal da Grande Dourados, Dourados - Federal do Paraná, Pato Branco – PR MS; 3 Universidade Federal do Mato Grosso do Sul, Campo Grande - MS; 4 Universidade Tecnológica Introduction: As known, medicinal plants are Federal do Paraná, Pato Branco – PR important source of drugs and some of them can cause harm, including DNA damage. Infusions from leaves Introduction: Medicinal plants, as other conventional of Alibertia edulis are popularly used in folk medicine drugs, have the same potential to cause harm, as it presents hypoglycemic, antihypertensive and including DNA damage. Infusions from leaves of diuretic effects. However, studies on the genotoxicity Alibertia edulis are popularly used in traditional of this plant are not available in the literature. medicine as it presents hypoglycemic, antihypertensive Objective: Investigate the possible mutagenic effect of and diuretic effects. However, studies on the the aqueous extract of Alibertia edulis (AEAE) in rats. genotoxicity of this plant are not available. Objective: Material and Methods: Sixty male and female Wistar Therefore, the purpose of this study was to evaluate the rats (8-12 weeks) were divided into six experimental frequency of damage and the damage index caused to groups (n=10). C+, as the positive control group, the DNA after 28 consecutive administrations of the received an intraperitoneal injection of aqueous extract of Alibertia edulis (AEAE) in rats. cyclophosphamide monohydrate (20 mg/kg i.p.) 48 Material and Methods: Sixty male and female Wistar hours before the euthanasia process. C-, as the negative rats (8-12 weeks) were divided into six experimental control group, received saline by oral gavage. Groups groups (n=10). C+, established as the positive control I, II, III and IV received four different doses of the group, received an intraperitoneal injection of AEAE (125, 250, 500 and 1000 mg/kg) by oral gavage cyclophosphamide monohydrate (20 mg/kg i.p.) 48 ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 173 hours before the euthanasia process. C-, as the negative micronucleus (CBMN) test in human blood cells. control group, received saline by oral gavage. Groups Materials and Methods: The in vitro CBMN assay in I, II, III and IV received four different doses of the human blood cells was performed according to the AEAE (125, 250, 500 and 1000 mg/kg p.o.) by oral OECD TG487 guideline. After 44 h of incubation, the gavage. Blood samples were collected by caudal treatment was carried out at 37ºC, with solvent-control puncture from each rat with a needle soaked in heparin (5% DMSO) and compounds tested in concentration and material was analyzed by fluorescence range from 150 to 10,000μM in 5% DMSO for 4 h, microscopy. This test was performed under the with positive controls mitomycin C 0.5μg/mL and approval of the Ethics Committee in Animal 1.0μg/mL for 2 h, and Colcemid 0.02μg/mL for 28 h. Experimentation from the Federal University of The analysis was performed under a fluorescence Grande Dourados (protocol: 29/2015). Results: The microscope (400X magnification) for the presence of negative control groups presented low frequency as micronucleus per 2,000 binucleate cells selected at well as low index of DNA damage. On the other hand, random by culture or experimental group. Results and cyclophosphamide monohydrate significantly Discussion: MGZ showed clastogenic and/or increased the frequency as well as the index of DNA aneugenic highly significant effect (p < 0.01) at damage to the tested cells when compared to the concentrations 1,600 and 4,000 µM. From the 7 negative control group. All doses of the AEAE were compounds analogous to MGZ studied, 5 showed statistically similar to the negative control group since negative results. Conclusions: Substitution of the they did not induce any significant increase of the thiadiazolyl ring of MGZ by the 1,2,3-triazole ring overall number of damaged cells nor the severity of the with cyclopropyl linked to triazole C-4 and the change cell damage. Conclusion: Data show that the AEAE of nitro group at 5-position to 4-position of the presented low genotoxicity at the doses tested in rats. imidazole ring abolished the mutagenic effect of MGZ. Financial support/Acknowledgments: CAPES and Compound 8 was the most promising of the analogous CNPq. compounds to MGZ studied because besides the absence of clastogenic and/or aneugenic effect, it showed high trypanocidal activity, being two times MC 18 - IN VITRO MUTAGENICITY TEST OF more potent than benznidazole and MGZ. NITROIMIDAZOLES AS A TOOL IN THE Financial support/Acknowledgments: Coordenação SEARCH FOR NEW TRYPANOCIDAL AGENTS de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); Fundação Oswaldo Cruz (FIOCRUZ). TROMPOWSKY, A.C.M. 1; CONDE, T.R. 1; CARVALHO, A.S. 2; QUARESMA, B.M.C.S. 2; BOECHAT, N. 2; ZAMITH, H.P.S. 1 MC 19 - ASSESSMENT OF DNA DAMAGE IN AGRICULTURAL WORKERS IN SOUTHERN 1. Department of Pharmacology and Toxicology, BRAZIL National Institute of Quality Control in Health (INCQS), Oswaldo Cruz Foundation (Fiocruz), Rio de BERNIERI, T. 1; MORAES, M. F. 1; ARDENGHI, P. Janeiro RJ, Brazil; 2. Department of Synthesis of G. 1; SILVA, L. B. 1. Drugs, Farmanguinhos, Fiocruz, Rio de Janeiro, RJ, Brazil. 1. Universidade Feevale, Novo Hamburgo-RS, Brasil.

Introduction: In terms of public health, Chagas disease INTRODUCTION: Agricultural workers represent a and sleeping sickness are considered important but population that is highly vulnerable to the toxic effects neglected human disorders, that is, those which have a of the pesticide exposure. The ability of pesticides to restricted therapeutic arsenal and most times with induce DNA damage has been reported in a number of undesirable high toxicity, including mutagenic studies and the findings suggested that exposure to potential. The Institute of Drug Technology pesticides can be genotoxic and carcinogenic to (Farmanguinhos), located at Oswaldo Cruz humans. OBJECTIVE: The aim of this study was to Foundation, has been developing projects of new evaluate the genotoxic effect of pesticides in molecules with possible trypanocidal activity, by agricultural workers, who live in the northwest of Rio synthesizing nitroimidazole compounds analogous to Grande do Sul, southern Brazil. METHODOLOGY: megazol (MGZ) using bioisosterism, in an attempt to All participants signed an informed consent document obtain drugs that cause lower incidence of side effects and completed a questionnaire in which they informed and no mutagenicity. The MGZ has potent their age, gender, occupation, contact with pesticides, trypanocidal activity but cannot be used clinically due smoking and alcohol habits. The exposed group to its mutagenicity. Objective: Evaluate if mutagenic consisted of soybean growers. The control group effect was abolished in 7 nitroimidazole compounds consisted of individuals employed in the fields of analogous to MGZ using in vitro cytokinesis-block administration, commerce, and education. For the ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 174 comet assay, a sample of whole blood was collected from each individual by a qualified professional. Samples were kept at 4 °C in the ice box and transported to the laboratory within 24 hours. For each subject, 100 cells were analyzed under light microscopy classifying them visually into five damage classes (0, I, II, III, and IV). The damage index was calculated by the sum of the number of nuclei multiplied by the value of the respective class, generating an index that ranges from 0 to 400. RESULTS: The exposed and control groups were composed of 12 subjects, all males. The average pesticide exposure time for farmers was 30.8 ± 15.2 years. The groups did not differ in terms of age (farmers 45.6 ± 15.4 years and control 42.3 ± 9.0 years; p = 0.98). The farmers have showed a significantly higher DNA damage index than the control group (118.4 ± 69.1 vs. 1.3 ± 1.7, p < 0.0001). CONCLUSION: The results indicate that farmers are exposed to mixtures of pesticides with genotoxic potential. Financial support/Acknowledgments: Feevale.

NANOTOXICOLOGY

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NT 01 - ARE BISMUTH NANOPARTICLES NT 02 - ENCAPSULATION OF ARTEMETHER SAFE TO US? CHARACTERIZATION AND IN PCL NANOCAPSULES: EFFICACY AND CYTOTOXICITY OF BISMUTH CARDIOTOXICITY EVALUATION NANOPARTICLES AND THEIR EFFECTS ON MAMMALIAN CELLS SOUZA, A.C.M.1; MOSQUEIRA, V.C.F.1; GARCIA, G.M.1; SILVEIRA, A.P.A.1; ANTUNES, L.R.1; REUS, TL1,3, MACHADO, TN2, PASCHOAL, GUIMARÃES, H.N.3; RICHARD, S.2; GRABE- ACC1, ABUD, APR1,3, KULIGOVSKI, C1,4, GUIMARAES, A.1. BEZERRA-JR, AG2, AGUIAR, AM1,3, DALLAGIOVANNA, B1,3 1 Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal de Ouro Preto, 1 Instituto Carlos Chagas – Fiocruz Paraná 2 Ouro Preto - MG, Brasil; 2 2Physiologie et Médecine Universidade Tecnológica Federal do Paraná 3 Expérimentale du Cœur et des Muscles - Universidade Federal do Paraná 4 Instituto de Biologia PHYMEDEXP, Inserm U1046, CNRS UMR 9214, Molecular do Paraná. Université de Montpellier - Montpellier, França; 3 Departamento de Engenharia Elétrica, Universidade Introduction: Nanoparticles (NPs) have a great Federal de Minas Gerais, Belo Horizonte - MG, Brasil. potential for technological and medical purposes. Among all types of NPs, bismuth NPs (BiNPs) are Introduction: Artemether (ATM) is an antimalarial very promising ones. BiNPs have bactericidal and drug. Its adverse effects and short half-life are a fungicidal properties, and they have also been limiting factor for malaria therapy. Aim: the objective evaluated for their application in imaging procedures. was to evaluate reduction of ATM loaded in poly-ε- However, so far, too little is known about their caprolactone nanocapsules (NC) cardiotoxic effects toxicity. Thus, a better comprehension about BiNPs and its efficacy against Plasmodium berghei. toxicity and also their effects on biological systems is Methodology: NC-ATM was obtained by interfacial required before they are used for biomedical purposes. polymer deposition. Male black c57BL6 healthy or Objective: Therefore, the main objective of this work infected with P. berghei received oral treatment with in to establishment of the toxic doses of BiNPs and vehicle; blank NC; NC-ATM and free ATM at 40, 80 also to understand how cell interacts with BiNPs. and 120 mg/kg, given 4 days, twice daily. Antimalarial Material and methods: BiNPs were synthesized by activity and mice survival were observed. laser ablation, stabilized with proteins from medium Electrocardiogram (ECG) lead II was monitored before (BiNPmedium) and bovine serum albumin (BiNPBSA) and after treatment. Results: The NC-ATM was shown and characterized by Dynamic Light Scattering, to be effective and safe. NC-ATM (120 mg/kg) Transmission Electron Microscopy (TEM) and Zeta reduced parasitemia and increased survival about potential analyzer. In order to assess the toxicity, 100%. The NC-ATM was also able to reduce the QT BALB/c 3T3 cells were used. Cell viability was and QTc interval prolongation about 46% and 38%, evaluated by neutral red uptake and IC20, IC50 and respectively, in healthy mice, at 24 hours after last IC80 values were obtained. Cell death was evaluated dose, compared to free ATM. In infected mice NC- by TUNEL assay. Interaction between cells-BiNPs ATM (120 mg/kg) was also able to reduce the was assessed by TEM. Results: BiNPs have an average cardiotoxic effect, reducing QT and QTc intervals size of 52 nm (BiNPmedium) and 49 nm (BiNPBSA). prolongation about 52% and 52%, respectively, They are also heterogeneous in size and rounded compared to free ATM. Conclusion: Free ATM was shaped. Their charge is -20,2 mV (BiNPmedium) and - effective but it was cardiotoxic to mice. ATM loaded 23,8 mV (BiNPBSA). For BiNPmedium, IC20, IC50 e in NC was more effective as antimalarial than free IC80 values were respectively: 78,32±18,90, form and the most important result is that NC-ATM 28,51±9,96 and 9,18±6,51 and for BiNPBSA, they was not cardiotoxic. were: 71,82±16,67, 25,54±8,37 e 8,60±3,35. Cell death Financial support/Acknowledgments: FAPEMIG occurred through apoptosis (TUNEL) and once BiNPs (Rede NanobioMG, PPM V 1227); CNPq; UFOP; were internalized, they led to cell damage and cell CAPES/COFECUB (738/13), CAPES, INSERM death (TEM). U1046. Financial support/Acknowledgments: CNPq, Fundação Araucária, CAPES, FIOCRUZ, Plataformas tecnológicas da FIOCRUZ.

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NT 03 - NANOTOXICITY OF SILVER NT 04 - CELL VIABILITY AND STABILITY OF NANOPARTICLES FROM BIOLOGICAL A NANOEMULSION ANTI-AGING SYNTESIS FORMULATION

PASCHOAL, A.C.C.1*; KULIGOVSKI, C. 1,4; REIS, TOSTA, F.V.1; NASCIMENTO, T.L.1; MOREIRA, G. 2; NAKAZATO, G. 2; PANAGIO, L.2; REUS, L.C.1; VALADARES, M.C.1; LIMA, E.M.1. T.L.3; DALLAGIOVANNA, B.1; ABUD, A.P.R.1; AGUIAR, A.M.1 Post-Graduation Program in Health Sciences 1FarmaTec - Center for Research, Development and 1 Laboratório de Biologia Básica de Células-Tronco, Technological Innovation in Drugs, Medicines and Instituto Carlos Chagas, Fiocruz, Curitiba, PR. Brazil; Cosmetics - Faculty of Pharmacy - Federal University 2 Universidade Estadual de Londrina, PR. Brazil; 3 of Goiás, Goiânia, Brazil. Universidade Federal do Paraná, PR. Brazil; 4 Instituto de Biologia Molecular do Paraná – IBMP, PR. Brazil Introduction: Nanoemulsions are extremely promising colloidal systems that can be used for several Introduction: Silver nanoparticles (AgNPs) are the applications. They used to be biocompatible and nanomaterials with the highest number of products on biodegradable systems in which the size, constituents the market, especially for their germicidal activity. Its and membrane surface can influence its effects, in synthesis occurs mainly in physicochemical form, particular cell viability. Objective: To develop and being economically and environmentally unfavorable. characterize a nanoemulsion formulation containing a As alternatives to renewable sources, fungal lipophilic cosmetic ingredient; test its stability and bioproduction has significant practical advantages. evaluate cell viability. Method: The nanoemulsion, However, little is known about the safety and toxicity consisting of fatty acids, phospholipid, nonionic of AgNPs on human and animal health, especially of surfactant, preservative, chelating agent, water and the the AgNPs from biological synthesis, making it lipophilic cosmetic agent, was obtained by the high necessary to search for effective methodologies to pressure emulsification and homogenization method. evaluate their toxicity. Objectives: The aim of this The formulation was characterized using mean work is to assess cytotoxic effect of biological AgNP- diameter, polydispersity index (PdI), zeta potential bio and chemical nanoparticles (AgNP-Che), to delimit (ZP), pH, cosmetic content and stability evaluation for safe concentrations and to determine if the toxic effect 30 days upon storage at 25 °C. Regarding cell varies as a function of the synthesis process. viability, 8 concentrations of the nanoemulsion (6x10- Methodology: The AgNP-Bio were biosynthesized by 4 to 10%) were prepared in DMEM medium the fungus Fusarium oxysporum in collaboration with supplemented with 2% FBS according to the ICCVAM the State University of Londrina (Patent, 2006, recommended protocol for the BALB/c 3T3 Neutral PI0605681). AgNP-Che were obtained commercially Red Uptake (NRU) cytotoxicity test. Results: Particles (SIGMA-ALDRICH® Saint Louis, USA). AgNPs stored at 25 °C had a mean diameter of 124 nm, PdI were characterized by UV/VIS spectrometry, Dynamic 0.1, ZP ‒50 mV, pH 8.3 and a cosmetic content of light scattering (DLS) and transmission electron 1325.21 µg/mL, not showing any change in these microscopy (TEM). To evaluate the cytotoxicity, the parameters during the study period. It was observed method of neutral red uptake on Balb 3T3 cells clone that, from the 4.10-2% concentration of formulation A31 was used (OECD 129). Results: TEM showed that the cell viability decreased to values below 75% after AgNP-Che is larger and more concentrated than 48h of treatment, and when incubated with the three AgNP-Bio. Both AgNPs have a spherical morphology highest concentrations of the formulation, cell viability and heterogeneous size. The DLS data corroborate the was close to 0%. The values for IC50 were 0.12% for microscopic observation, showing the heterogeneity blank nanoemulsion and 0.11% for the nanoemulsion and reporting an average size for the 97nm AgNP-Che containing the cosmetic ingredient. Conclusion: The and 56nm AgNP-Bio. Through the cytotoxicity test, nanoemulsion formulation showed stability for as a AgNP-Che presented IC50 values of cosmetic ingredient. The IC50 value will serve as the 76.43±13.24μg/mL and AgNP-Bio presented IC50 limiting concentration for efficacy trials. values of 1.41±0.21μg/mL. Conclusion: The data Financial support/Acknowledgments: CAPES and presented indicate a difference in toxicity between CNPq. AgNP-Che and AgNP-Bio, the latter being apparently more toxic, indicating a possible toxicological profile varying as a function of the synthesis. New trials will be carried out with new batches of AgNP-Bio besides the evaluation of the mechanisms of action involved. Financial support/Acknowledgments: CAPES, Fundação Araucária, CNPq, Fiocruz. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 178

NT 05 - CYTOTOXICITY EVALUATION OF with improve the methods of dispersion of these FULLERENE AND TITANIUM OXIDE nanoparticulate materials. NANOPARTICLES Financial support/Acknowledgments: CNPq / CAPES / FINEP. BRITO, H. R. G. 1; BERNARDES, A. P. 2; BEZERRA, S. F. 1; LIMA, E. M. 2; SILVA, A. L. P. 1; VALADARES, M. C. 1 NT 06 - THE USE OF OECD TG 432 TO EVALUATE DE PHOTOTOXICITY OF NON- 1. Laboratório de Farmacologia e Toxicologia Celular NANOMETRIC AND NANOMETRIC – LFTC, Faculdade de Farmácia, Universidade Federal PARTICLES OF ZINC OXIDE de Goiás, Goiânia-GO, Brasil; Laboratório Nanotecnologia Farmacêutica e Sistema de Liberação SILVA, J. V. M. A. 1; OLIVEIRA, J. F. 1; SOUZA, Controlada de Fármacos - FARMATEC, Faculdade de V. 1; BALOTTIN, L. B. L. 1 Farmácia, Universidade Federal de Goiás, Goiânia - GO, Brasil. 1. Instituto Nacional de Metrologia, Qualidade e Tecnologia, Duque de Caxias -RJ, Brasil. Introduction: Fullerene C60 and titanium oxide nanoparticles (TiONPs) have been widely used in Sunscreens produced with nanoparticles of ZnO have technological applications, including electronic superior UV reflection and absorption capacity and engineering, pharmaceuticals, cosmetics, industrial, transparency in visible light, preventing the formation consumer products due to their mechanical, electrical, of undesirable white film on the skin. However, there optical, magnetic, biological properties and to their is no consensus in scientific community on the strong catalytic activity. Objective: Investigate the nanomaterials toxicity, mainly due to the use of cytotoxic effects of C60 fullerene and TiONPs on methodologies that do not allow comparability. So, the fibroblasts cells of Balb / c 3T3 mice, as well as to aim of this study is to use the internationally perform the characterization of these substances. harmonized methodologies to evaluate using in vitro Methods: Evaluation of the cytotoxicity was performed strategy the impact the size distribution of ZnO to by BALB/c 3T3 Neutral Red Uptake Cytotoxicity phototoxicity potential. The non-nanometric and Assay (3T3 NRU), (OECD 129). The cells were nanometric ZnO was disperse by ultrasonication and cultivated according to the protocol and exposed to size distribution evaluate by Dynamic Light Scattering, different concentrations of the substances test for 48 following ISO 22412:2008 and NIST guidelines, and hours. Size and zeta potential of the nanoparticles were phototoxicity evaluated according the OECD Test determined by Dynamic Light Scattering (DLS) and Guideline nº 432. This OECD test compare the electrophoretic mobility, respectively. C60 fullerenes reduction of cell viability of Balb 3T3 cells, measured were treated with Tetrahydrofuran (THF) under by neutral red uptake, in presence and absence of solar stirring for 12h. Equal volume of water was added and simulated light (1,7 mW/cm² for 50 min). Through PIF THF was removed under vacuum. TiONPs were or MPE score, the test substance can be classified in dispersed in 95% ethanol overnight, and the same phototoxic, probable phototoxic and no phototoxic. In- protocol for the removal of the organic solvent was house proficiency of OECD TG nº 432 was achieved used, followed by 10 minutes of Ti probe sonication. using two positive, chlorpromazine and anthracene, Nanoparticle dispersions were measured for size and and two negative controls, sodium lauryl sulfate and size distribution in a ZetaSizer NanoS from Malvern hexachlorophene. By different conditions of Instruments and for zeta potential in a Zetaplus® from ultrasonication dispersion, we established three Brookhaven Results: For both types of nanoparticles, independent populations of ZnO in DLS (n=6): Z1ave agglomerates were observed even after sonication, = 421,9 nm ± 13,6 nm and PDI1 = 0,346 ± 0,035; highlighting the need to further reduce the surface Z2ave = 204,8 nm ± 3,6 nm and PDI2 = 0,326 ± 0,018; tension of these nanoparticles in order to examine their Z3ave = 101,0 nm ± 0,65 nm and PDI3 = 0,171 ± individual behavior. Zeta potential measurements 0,019. For Z1, PIF = 2,579 ± 0,073 and MPE = 0,090 ± evidenced instability of the dispersions. In these 0,009; the population Z2, PIF = 2,711 ± 0,450 and samples, no cytotoxic effects were observed in all MPE = 0,110 ± 0,005; and Z3, PIF = 3,005 ± 0,156 concentrations investigated. The presence of these and MPE = 0,182 ± 0,013. No significant difference agglomerates in the biocompatible vehicle could be (p=0,2376) when evaluated the PIF score in three influenced the results of the cytotoxicity assay. populations, classifying as probable phototoxic. Conclusion: The substances evaluated were not Though, when considered the MPE score, the p < cytotoxicity. Further testing at higher concentrations 0,0001 show significant difference and, as far as the and/or with repeated exposure time is required to size decrease, the classification pass to no phototoxic verify the toxic effects of these nanomaterials along to phototoxic. The results suggest that the only international validated method to evaluated the ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 179 phototoxicity is not enough to classified the different of nitrite by peritoneal macrophages; however, no sizes of ZnO, so the use other non-validated test is differences were observed when the cells were necessary. stimulated. Conclusions: Our results demonstrate that Financial support / Acknowledgments: successful translation of nanomaterials to medicine Inmetro/Renama/CNPq. depends on its toxicological profile. In addition, an environment of stimulated cells and heterogeneous phenotypes must be considered for the immunotoxicity NT 07 - INVESTIGATION OF POTENTIAL assessment during nanomaterials development. CLINICAL IMPLICATIONS OF A HYBRID Financial support/Acknowledgments: CNPq GRAPHENE-SILVER NANOMATERIAL: A 140560/2014-9, CNPq/MCTIC NanoBioss, FAPESP SAFE-BY-DESIGN APPROACH 15/23767-0.

DE LUNA, L. A. V. 1.2; ZORGI, N. E. 2; DE MORAES, A. C. M. 1; GIORGIO, S. 2; ALVES, O. L. NT 08 - ACUTE TOXICITY EVALUATION IN 1 VIVO AFTER EXPOSURE TO COPPER OXIDE NANOPARTICLES (1) Laboratory of Solid State Chemistry (LQES), Institute of Chemistry (IQ), University of Campinas MACIEL MAGALHÃES, M. 1; SAVIGNON, T. 1; (UNICAMP), Campinas, Brazil. (2) Laboratory of MEDEIROS, R. J. 1; SU, T. C. S. 1; AMENDOEIRA, Leishmaniasis and Cell Culture (Lableish), Institute of F. C. 1 Biology, University of Campinas, Campinas, Brazil. 1. Instituto Nacional de Controle de Qualidade em Introduction: Graphene has been the most studied Saúde - FIOCRUZ - Rio de Janeiro – RJ. nanomaterial since its discovery by Geim and Novoselov in 2004. Graphene consists of Introduction: Copper oxide nanoparticles (CuO-NPs) bidimensional sheets of carbon atoms arranged in have been increasingly used in products of human hexagonal rings. Its highly oxidized form is the so- interest, such as antimicrobial coatings, lubricants and called graphene oxide (GO). Our group developed a cosmetics. Studies of CuO-NPs toxicity in in vitro hybrid nanomaterial based on graphene oxide and models are found in literature, as in a 2014 study, silver nanoparticles (GOAg) with high microbicide which shows that they have a higher toxic capacity activity and potential use in nanomedicine. Objective: than micro particles of same material to human To study the immunotoxicity elicited by this hybrid mesenchymal stem cell cultures. Since these can be nanomaterial, we performed in vitro and ex vivo used as adjuvants in many products, including analysis to survey several endpoints. Material and medicines and cosmetics, toxicity study of these methods: The cells were macrophages obtained from nanoparticles is relevant to determine safe levels of use BALB/c mice peritoneum and lineage J774. Ranges of in products intended for the population. Objectives: concentrations from 1 to 25 µg mL-1 were tested. The Evaluate the acute toxicity of CuO-NPs; Verify apoptosis and the mitochondrial depolarization were hematological and biochemical parameters changes; evaluated by flow cytometry. The lipid peroxidation Analyze histopathologically the following animal’s was evaluated in a High-Content Imaging system. For organs: brain, spleen, liver and kidneys. Materials and the assessment of GOAg inflammatory potential, methods: Wistar female rats were used, with 8 to 12 peritoneal macrophages were stimulated with IFN-γ weeks of life, nulliparous and not pregnant at the time and production of IL-1β, TNF-α and IL-10 cytokines of administration (Oswaldo Cruz Foundation Ethics was determined by the Cytometric Bead Array method. Committee register number 11/2017-1). CuO-NPs Nitrite production was determined by the Griess were suspended in corn oil at three different doses - reagent after peritoneal macrophages stimulation with 50, 300 and 2000 mg.kg-1 -, and administered orally IFN-γ and LPS (bacterial lipopolysaccharide), through a gavage procedure using an appropriate followed by exposure to GOAg. Results: 5 µg mL-1 cannula for animals’ sizing. Control animals were induced 16 % of apoptosis and 32 % of late treated only with corn oil. This procedure was based apoptosis/necrosis after 24 h of exposure. The on OECD 420 guideline. After administration, mitochondrial depolarization was dose-dependent and parameters such as amount of food consumed and ranged from 8 to 22 %. The lipid peroxidation was weight gain were obtained daily, always at the same time and dose-dependent, and reached 781 % when time of day. At the end of 14 days of observation, the compared to non-treated cells. The GOAg did not animals were euthanized and had blood and organs induce peritoneal macrophages to produce the collected for hematological, biochemical and investigated cytokines after 48 h, however when the histopathological analyzes. Preliminary results: cells were stimulated with IFN-γ, there was significant Among all evaluated parameters, food intake in the production of IL-1β. GOAg increased the production first 3 days after CuO-NPs administration was the only ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 180 one that presented statistically significant variation, up and i.p) did not modify the body and organs weight, to the present moment. Conclusion: With present biochemical parameters and oxidative markers when results is possible to speculate that CuO-NPs may be compared to control groups. The micronucleus test did relatively safe, in cases of single exposure, for use in not identify differences between the groups. However, products intended for the population. However, other the higher dose (10 mL/kg) by intraperitoneal pathway toxicological tests, such as those for chronic and causes changes in hematological parameters, with a reproductive toxicity, are still needed to determine the significant increase in plasma proteins, platelets, total limits of its use. leukocytes, band cells and lymphocytes. In summary, Financial support / Acknowledgments: we can suggest that the oral route is safe at the doses INCQS/FIOCRUZ; INMETRO administered, since no signs of toxicity were observed. The changes in some hematological parameters observed with the higher doses of NE can indicate its NT 09 - IN VIVO TOXICITY EVALUATION OF potential toxicity. This study contributes to the LIPID NANOCARRIERS FOR DRUG knowledge on the safety and toxicity of nanocarriers DELIVERY with pharmaceutical application. Financial support/Acknowledgments: CNPq, HORT, M.A.1,2; ALVES, B.S. 1,2; RAMIRES- CAPES. JÚNIOR, O.V. 1,2; FALKEMBACH, M.C.1; ARAÚJO, G.M.S.1,2; FERNANDES, C.L.F1,2; BIDONE, J.3; DORA, C.L.1,2; DA SILVA-JÚNIOR, F.M.R. 1,2

1 Instituto de Ciências Biológicas, Universidade Federal do Rio Grande, Rio Grande, RS, Brazil 2 Programa de Pós-graduação em Ciências da Saúde, Faculdade de Medicina, Universidade Federal do Rio Grande, Rio Grande, RS, Brazil 3 Departamento de Farmácia, Centro de Ciências Químicas, Farmacêuticas e de Alimentos, Universidade Federal de Pelotas, Pelotas, RS, Brazil.

Lipid nanoparticles are an emerging tool for drug delivery, due their ability to incorporate lipophilic drugs, protect the drug from degradation, improve bioavailability and control release. The pharmaceutical potential of lipid nanocarriers has been extensively studied; however the possible toxic effects of these systems have not been sufficiently investigated thus far. Thus, the aim of this study was to evaluate the possible toxic effects of nanoemulsions (NE) in vivo. NE were prepared with medium-chain triglyceride, lecithin E80, solutol and PBS by hot emulsification followed by high pressure homogenization. Male Wistar rats (2 months - 250 g) were treated once daily for 21 days with NE by oral (o.p.) or intraperitoneal pathway (i.p.) at the doses of 10, 5 or 2.5 mL per kg (n = 6/group) or physiological solution (control groups). At the end of the experiment rats were killed, their blood was collected to biochemical and hematological analysis, and selected organs were removed and weighted. Oxidative stress markers (lipoperoxidation and non-protein thiols) were measured in the liver. Mutagenicity was evaluated via the micronucleus test in cells from the bone marrow. Body weight was evaluated every two days and at the end of treatment. The experimental protocol was approved by the Institutional Ethics Committee (23116.008166/2015- 67). Our results show that the treatment with NE (o.p

OCCUPATIONAL TOXICOLOGY

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 182

OC 01 - METHOD VALIDATIONFOR concentrations of 1.5, 2 and 2.5 μg are adequate to QUANTIFICATION OF GLOBAL DNA evaluate global methylation levels. METHYLATION BY HPLC-DAD IN Financial support / Acknowledgments: Universidade DIFFERENT DNA CONCENTRATIONS Feevale; UFRGS.

SANTOS, A. L. A; BARBOSA, E. 1; ANTUNES, M. 1; ROVARIS, D. 2; BAU, C. 2; LINDEN, R. 1; OC 02 - TOXICOLOGICAL AND BEHAVIORAL CHARÃO, M. 1 PROFILE OF TRUCK DRIVERS ON A ROAD OF THE WEST-CENTER REGION OF BRAZIL 1 Laboratório de Análises Toxicológicas, Universidade Feevale, Novo Hamburgo - RS, Brasil; 2 Instituto de COSTA, C. D. D1; D.S. RODRIGUES 1, F.N. MEIRA Biociências, Departamento de Genética, Universidade 2, L.C. CUNHAA, S.H.N. COSTA1, V. RODRIGUEZ Federal do Rio Grande do Sul, Porto Alegre - RS, 3, K.C. ALCÂNTARA 1. Brasil. 1 Universidade Federal de Goiás, Goiânia, GO, Brasil. INTRODUCTION: Nowadays there is a great concern 2 Secretaria de Estado da Saúde do Distrito Federal, about the exposure to a different set of chemicals Brasília, DF, Brasil. 3 Pontifícia Universidade Católica which may lead to changes in the epigenome. An easy de Goiás, Goiânia, GO, Brasil. way to identify epigenomic changes is to quantify the global methylation of DNA using high pressure liquid Background: The cargoes transportation is a very chromatography (HPLC) to quantify 5-Methyl-2’- important activity to the Brazilian economy where deoxycytidine (5mdC) and 2'-deoxycytidine (dC) there are about 1.1 million truck drivers. A lot of them levels. OBJECTIVE: This study aims to develop a are related to accidents on the roads and to the use of reliable method to assess dC and 5mdC levels using psychoactive substances. Objective: The aim of this HPLC with diode array detector.MATERIAL AND study was to verify the toxicological and behavioral METHODS: The DNA was extracted from whole profile among truck drivers. Methods: A total of 697 blood. Concentrations of 1, 1.5, 2 and 2.5 μg of DNA driver’s trucks were interviewed at a gas station on were treated with Rnase A and T1. Samples were BR-153 located in the West Center region of Brazil, incubated with Dnase I and nuclease P1 at 37 °C. between February 2014 and February 2015. The Alkaline phosphatase was added to the samples which ASSIST questionnaire was applied to screening the use were incubated overnight.Samples were centrifugated of alcoholic beverages, tobacco and illicit drugs as at 14,000 rpm, 5 minutes, and the supernatant was cannabinoids, cocaine, amphetamine or ecstasy, injected on an HPLC-DAD. Separation was performed inhalants, hypnotics/sedatives, hallucinogens, opioids. in a C8 column (150x4.6 mm, 5 µm) at 20 °C. The Sociodemographic information (age, sexual nucleotides were eluted using a gradient mobile phase orientation, marital status, education, region of with 25 mM KH2PO4 and 0.1% acetic acid (A) and residence), professional characteristics (origin and methanol (B), at flow rate of 0.6 ml/min, and truck cargoes destination, time away from home) and monitored at 280 nm. The relative content of 5mdC information about previous sexual transmitted was expressed as a %5mdCin relation to the total infections were collected. Basics descriptive statistics amount of cytosine (dC + 5mdC).It was evaluated the were performed using the software Epi Info. The DNA amount required for the analyses of the ASSIST answers were done by adding methodology.RESULTS:The retention time for dC was scores of questions. To analyses the association 6 and for 5mdC was 9 minutes. The total analytical run between the variables was used multiple time was 30 minutes. The calibration data presented correspondence analysis and hierarchical cluster heteroscedasticity, Fexp = 751.10 a 6082.15 (Ftab (5; analysis. Results: All of the participants were male 5; 0,95) = 5,05). Among the evaluated weighting with a median age of 42 years old, most of them were factors, 1/x presented the smallest % relative error. married and have low scholarity. 47% related moderate Calibration curves range from 0.09 to 6.25 µg/mL or high risk of consumption of alcohol and tobacco, (r=0.99) for dC, and from 0.011 to 0.78 µg/mL (0.99) respectively and 21% used of cocaine, cannabinoids, for 5mdC. Precision assays presented CV% lower than amphetamine, hypnotics/sedatives, opioids, inhalants 5.2% for dC and 3.5% for 5mdC. Accuracy was 98.8– and needed to go through a brief intervention or are be 100.5% for dC and 92.4-101.2% for mdC. The advised to seek specialist treatment for these drugs, %5mdC levels were 3.77, 8.2, 7.7 and 7.9% for the respectively. Younger and single truck drivers showed DNA concentrations of 1 to 2.5 μg, high tobacco, cannabis and amphetamines respectively.CONCLUSION:A reliable method for the consumption and needed to receive brief intervention quantification of dC and 5mdC in whole blood samples for these drugs. Conclusions: Brazilian truck drivers was developed. The procedure has an adequate interviewed related high use of tobacco, alcohol, analytical performance. The results show that DNA cocaine and other ilícit drugs. Measures are needed to ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 183 improve quality of life to the drivers, as well as to There were 26 inhibitions for ACHE and 12 for prevent and treat individuals who use licit and illicit BCHE.The inhibitions averages were 3.93% ± 2.9 and drugs in order to reduce engagement with traffic 7.84% ± 2.1 for ACHE and BCHE, respectively. accidents. Inhibition >20% predominated for ACHE. One Financial support / Acknowledgments: hundred and seventy one EWs used just one insecticide Departamento de DST, Aids e Hepatites Virais da of which 65.5% reported Pyriproxifen; 20.5% Secretaria de Vigilância em Saúde Malathion and 14% Bendiocarb.The use of combined (DDAHV/SVS/MS), Escritório das Nações Unidas insecticides was lower (87 EWs) and 67.8% of the sobre Drogas e Crime (UNODC), Coordenação de EWs used up to 2 insecticides. Malathion and Aperfeiçoamento de Pessoal de Nível Superior Bendiocarb association was the most reported (74.7%). (CAPES). Conclusion: The analyzes were performed in all mesoregions of Santa Catarina.The VI had the most expressive participation. The majority of EWs OC 03 - BIOLOGICAL MONITORING OF performed baseline analysis.In most applications, a ENDEMIC CONTROL WORKERS OF THE single insecticide was used and Pyreproxyfen STATE OF SANTA CATARINA prevailed. A higher number of EWs exhibited inhibition for ACHE while BCHE was more SILVA, C.I.1; VANDRESEN, B.1; PACHECO, inhibited.The biological monitoring is important since L.K.1; FERREIRA, C.2; PRADO, M.M.T.3; DOS no EW showed inhibition higher than the maximum SANTOS, C.R. 1,4. biological index, despite observed inhibitions. Financial support / Acknowledgments: Secretaria de 1.Divisão de Análises Clínicas, Hospital Universitário Estado da Saúde, Universidade Federal de Santa Professor Polydoro Ernani de São Thiago, Catarina Universidade Federal de Santa Catarina, Florianópolis- SC, Brasil; 2. Laboratório Central de Saúde Pública de Santa Catarina, Florianópolis- SC, Brasil; 3. Diretoria OC 04 - A STUDY OF THE RELATIONSHIP de Vigilância Sanitária da Secretaria de Estado da BETWEEN OCCUPACIONAL EXPOSURE TO Saúde de Santa Catarina, Florianópolis- SC, Brasil; 4 FORMALDEHYDE AND THE FORMIC ACID Departamento de Patologia, Centro de Ciências da LEVELS IN URINE OF BEAUTY SALON Saúde, Universidade Federal de Santa Catarina, WORKERS Florianópolis- SC, Brasil. BARBOSA, E. 1; SANTOS. A. L. A. 1; PETEFFI, G. Introduction: Cholinesterase inhibitors are some of the P. 1; ANTUNES, M. V. 1; LINDEN, R. 1; CHARÃO, insecticides used by Endemic Control Workers (EW) M. F. 1. for the control of Aedes aegypti. Cholinesterase activity is the biological indicator recommended. 1 Laboratório de Análises Toxicológicas, Universidade Objective: To describe the results of biological Feevale, Novo Hamburgo, Rio Grande do Sul, Brasil. monitoring of EWs and the profile of pesticides used in mesoregions of Santa Catarina INTRODUCTION: Currently in Brazil there is a Methodology:Prospective descriptive study of widespread use of formaldehyde (FA) as a hair acetylcholinesterase (ACHE) and smoothing agent which is illegaly added to maximize butyrylcholinesterase (BCHE) activities of EWs the smothing effect of hair smoothing creams. analyzed by Toxicology Laboratory (TL) at the Consequently many beauty salon workers are University Hospital, in partnership with the Public frequently exposed to FA and its effects. OBJECTIVE: Health Central Laboratory between February/2016 and This study aims to assess and find the correlation July/2017.Results: The TL conducted 966 analyzes among formaldehyde levels in the air and smoothing related to 304 EWs from 34 counties. Vale do Itajaí products, and the levels of formic acid in urine of the mesoregion (VI) evaluated 152 EWs and 60.1% of the exposed workers and the control group. MATERIAL analyzes. The samples for ACHE and BCHE were AND METHODS: The exposed group was composed colleted at the same time and 73.5% of the EWs of 36 workers from 7 different beauty salons and the performed only baseline analysis. The mean values control group was composed of 30 volunteers. Seven were 483.2 ± 48.6 U.mmolHb-1 and 13342.2 ± 2895.3 different straightening product samples were analyzed U.L-1 for ACHE and BCHE, respectively. Seventy according to Wu et al. (2003) by high pressure liquid two EWs from 9 counties performed ≥2 collections. chromatography with diode array detector (HPLC- For the evaluation of the inhibition, 58 EWs were DAD) using a C18 octadecylsilane column and included, totaling 232 analyzes from 6 counties. The monitored in 345 nm. Umex-100 passive samplers mean time between collections was 3 months (± were used for 8 hours to assess the formaldehyde 2).Enzymatic inhibition was observed in 35 EWs. levels in the volunteers breathing zone and in the ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 184 salons. These passive samplers were analyzed by results. Mean activity of the erythrocyte cholinesterase HPLC according to Occupational Safety and Health enzyme in the rural workers presented a significant Agency method 1007 (OSHA, 2005) using a C18 decrease, indicative of inadequate exposure to column and monitored in 365 nm. Urinary formic acid cholinesterase inhibiting insecticides. Mean levels were assessed by gas chromatography with concentrations of the metals As, Cd, Mn, and Zn were flame ionization detector according to Peteffi et al. higher in rural workers compared to urban residents, (2015). RESULTS: The FA content in the product which suggests that inadequate exposure to fertilizers samples ranged from 3.35 to 6.45%. The passive and agricultural chemicals may result in their samplers used in the salons showed exposures ranging absorption. The blood counts did not present any from 0.028 to 0.24 ppm. Mean formate levels were changes, indicating that its use for monitoring 25.95 mg/L (SD=15.04) in the exposed group and occupational exposures to fertilizers and agricultural 17.64 mg/L (SD=8.81) in the control group (p < 0.05). chemicals is inadequate. We found strong correlations between the saloon FA Financial support / Acknowledgments: FCFRP - levels and the products FA content (r=0.868; p < USP. 0.001), but a weak correlation between individual FA exposure and urinary formic acid levels after exposure (r=0.388; p < 0.05). CONCLUSION: All the creams OC 06 - OCCUPATIONAL EXPOSURE TO had concentrations of FA higher than the 0.2% allowed CHROMIUM, NICKEL AND COBALT IN by ANVISA and the air levels of FA were higher than DENTAL PROSTHESIS LABORATORIES the 0.016 ppm exposure limit recommended by the National Institute for Occupational Safety and Health. ROCHA, G. H. O.; HOELTGEBAUM, D. 2; LINI, Therefore the salon workers were exposed to R.S. 2; BARBOSA, F. 3; SOUZA, V.C.O. 3; pottentialy harmful FA levels. Moreover, the use of BATISTA, B.L. 3,4; NERILO, S.B. 2; BANDO, E. 2; urinary formic acid levels to assess FA exposure NISHIYAMA, P. 2; MOSSINI, S.A.G. 2. remains unreliable. Financial support / Acknowledgments: 1 Faculty of Pharmaceutical Sciences, Department of FAPERGS/PPSUS. Clinical and Toxicological Analyses, University of São Paulo, São Paulo – SP, Brazil; 2 Laboratory of Toxicology, Department of Basic Health Sciences, OC 05 - HEMATOLOGICAL EVALUATION, State University of Maringá, Maringá – PR, Brazil; 3 ENZYME ACTIVITY AND LEVELS OF Laboratory of Metal Toxicology and Essentiality, METALS IN OCCUPATIONAL EXPOSURE TO Department of Clinical, Toxicological and DEFENSIVE AND AGRICULTURAL Bromatological Analyses, Faculty of Pharmaceutical FERTILIZERS Sciences of Ribeirão Preto, Ribeirão Preto - SP, Brazil; 4 Natural and Human Sciences Centre, Federal SARAIVA, E. R. 1; BORGES, D. 2; QUEIROZ, R. H. University of the ABC, Santo André – SP, Brazil. C. 3. Introduction: metallic alloys are among the most commonly used materials by dental prosthesis 1 Universidade de Rio Verde, Rio Verde - GO, Brasil; technicians. These are employed in the manufacture of 2 Universidade de Rio Verde, Rio Verde - GO; 3 dental restorations, removable prostheses, implants, Universidade de São Paulo, Ribeirão Preto - SP, orthodontic appliances, among others. Such alloys Brasil. consist, generally, of 35-65% cobalt, 20-30% chromium and 0-30% nickel. Dental prosthesis Advancements in the agrochemical industry associated technicians are very likely to be exposed to metal- with new seeding techniques make it possible for the containing dusts and vapors released during the country to reach annual crop records in production. production of dental prostheses, which have the However, the chemicals and fertilizers used for potential to cause adverse health effects. Robust increasing productions are by no means harmless to analytical techniques of great sensitivity and human health. Repeated and inadequate exposures to specificity, such as dynamic reaction cell inductively such chemicals may result in their absorption and coupled plasma mass spectrometry (DRC-ICP-MS), cause acute and chronic poisoning. In order to assess have been extensively used in recent years for analysis these exposures, we measured the activity of the of metals. Biomonitoring and continuous assessment acetylcholinesterase enzyme and evaluated blood of exposure to metals have been proven reliable counts. In addition, we measured blood levels of practices in the evaluation of occupational exposure of metals such as arsine (As), cadmium (Cd), lead (Pb), specific populations to such metals. Aims: to evaluate manganese (Mn), zinc (Zn), copper (Cu), and the occupational exposure to metals in dental prosthesis selenium in farm workers and urban residents in a laboratories workers by assessing chromium, nickel region of Rio Verde- GO- Brazil and compared the and cobalt in biological samples. Methods: a total of ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 185

68 workers from dental prosthesis laboratories in a city The average age was 51±14 to (1) and 48 ±15 years in southern Brazil were enrolled in the study. In serum old to (2). Regarding exposure, 53 farmers claimed and urine samples collected, levels of metals they apply pesticides (1A) and the other 25 farmers chromium, nickel and cobalt were assessed by DRC- work on the farm but do not apply (1B). The use of CI ICP-MS, which were compared to reference values was reported by 28% of the farmers corresponding to previously determined from 136 healthy, non-exposed 11% of the total pesticides used. The average of ACHE individuals. Results: chromium, nickel and cobalt were activity was for (1A), (1B) and (2), respectively, 498, found at higher levels in serum and urine of the 497 and 503 U.mmol Hb-1; while for BCHE the assessed workers when compared to reference values activity was 13,913, 14,856 e 15,473 U.L-1. Despite (p < 0.05). Gender and time of exposure did not not being statistically significant the results showed influence metal levels significantly. Conclusions: lower BCHE activity in 1A group. The activity was workers from dentistry laboratories are more evaluated by the use (3) or not use (4) CI. The results susceptible to exposure to metals due to use of metal- observed were for ACHE 503.26 (3) and 496.37 (4) containing substances in their workplace than the U/mmol.Hb-1 and for BCHE 13,386 (3) and 14,309 general population. Changes in work practice are U.L-1 (4). Conclusion: All the results were in the needed in order to prevent work related diseases and reference values. Although only one analysis was possible intoxications. made, it is possible to observe that the activity value of BCHE is likely to be reduced in the groups considered Financial support / Acknowledgments: We more exposed. Thus, new measurements in these acknowledge Fundação Araucaria for its financial groups should be analyzed in a way to confirm the support. tendency observed.

Financial support / Acknowledgments: OC 07 - EVALUATION OF FUNDAÇÃO DE AMPARO À PESQUISA E ACETYLCHOLINESTERASE AND INOVAÇÃO DO ESTADO DE SANTA CATARINA BUTIRYLCHOLINESTERASE ACTIVITY IN (FAPESC); MINISTÉRIO DA SAÚDE, MS; FARMERS FROM ANTÔNIO CARLOS – SC CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E SPEZIA, I.A. 1; SILVA, C.S. 2, PACHECO, K.L. 2, TECNOLÓGICO, CNPq; SECRETARIA DE FEIJÓ, A.J.L. 2, ZANNIN, M. 1,3, DOS SANTOS, ESTADO DA SAÚDE, SES-SC; MINISTÉRIO C.R. 2,3. PÚBLICO DO TRABALHO DE SANTA CATARINA (MPT-SC). 1 Centro de Informações Toxicológicas de Santa Catarina, Florianópolis - SC, Brasil; 2 Setor de Toxicologia, Divisão de Análises Clínicas, Hospital OC 08 – PATHOLOGIES AND USE OF Universitário Professor Polydoro Ernani de São MEDICATION IN FARMERS OF ANTÔNIO Thiago, Florianópolis - SC, Brasil; 3 Departamento de CARLOS – SANTA CATARINA Patologia, Centro de Ciências da Saúde, Universidade Federal de Santa Catarina, Florianópolis - SC, Brasil. Spezia, I. A. 1; SILVA, C.S. 2; PACHECO, K.L 2; FEIJÓ, A.J.L. 2; ZANNIN, M. 1,3; DOS SANTOS, Introduction: Acetylcholinesterase (ACHE) and C.R 2,3. butirylcholinesterase (BCHE) are biomarkers used to monitor the exposure to cholinesterase inhibitors (CI) 1 Centro de Informações Toxicológicas de Santa according the Brazilian Labor Ministry. Objective: Catarina, Florianópolis - SC, Brasil; 2 Setor de Evaluate the cholinestarase activity in farmers, under Toxicologia, Divisão de Análises Clínicas, Hospital different exposure conditions in the city of Antônio Universitário Professor Polydoro Ernani de São Carlos, in Santa Catarina. Material and methods: The Thiago, Florianópolis - SC, Brasil; 3 Departamento de prospective cohort study was conducted with Patologia, Centro de Ciências da Saúde, Universidade individuals questionnaires in their residences, followed Federal de Santa Catarina, Florianópolis - SC, Brasil. by blood collection. All analyses were performed in the Toxicology Laboratory at the Hospital of the Introduction: Many studies show that pesticides Federal University of Santa Catarina (UFSC), by using exposure has association with incidence presence of validated spectrofhotometrics methods. The study was some pathologies, even in occupational or approved by the Ethics Committee in Human Research enviromental exposure. Cancer, diabetes mellitus, of the UFSC. Results: 97 individuals were included, hypertension and mental disorders are examples of divided in two groups: (1) 78 farmers and (2) 19 non those problems. Objective: Evaluate the prevalence of farmers. Regarding gender, in (1) 42 and 52% and in pathologies and characterize the use of medication in (2) 16 and 84% were women and men, respectively. farmers at the city of Antônio Carlos, Santa Catarina. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 186

Methods: The prospective cohort study was conducted Introduction: Exposure to pesticides can be extremely through the application of questionnaires to individuals dangerous for humans and other living organisms. In in their residences. The study was approved by the the last decade, oxidative stress has been the focus of Ethics Committee in Human Research of the Federal several studies aimed at determining the association University of Santa Catarina. Data were evaluated with the toxic effects caused by these compounds. using Epidata 3.1 and SPSS 20.0. Results: Were Objective: The present study was conducted to included 97 individuals, divided in two groups: (1) 78 evaluate the oxidative stress among workers exposed farmers (42% of women and 58% men), and (2) 19 to fungicides. Materials and Methods: 58 rural workers non-farmers with 16 % women and 84% men). The who reported exposure to fungicides (last 30 days) and average age was 51.5±14 years old to (1) and 30 control subjects (not occupationally exposed to 48.1±14.9 years old to (2). Regarding exposure, 53 pesticides) were recruited. Oxidative stress biomarkers farmers affirmed the application of pesticides (1A) and were measured in plasma samples. Malondialdehyde the other 25 farmers work on the farm but do not apply (MDA) was analysed by high performance liquid pesticides (1B). The pesticides more cited were chromatography (HPLC) and oxidized proteins (OP) paraquat (11%), cholinesterase inhibitors (11%) and were determined spectrophotometrically. All results glyphosate (9.5%). The pathologies showed the were expressed in units per mg of total proteins, which following frequency: diabetes 9.5%, 8% and 10%, were measured using Bradford assay. According to cancer 6%, 8% e 10%, and thyroid disfuction 7%, 20% questionnaires, the fungicides used were triazoles and and 5%, to 1A, 1B and 2, respectively. Self-medication strobilurins. Cigarette smoking and ethanol use were was report by 43% (1A), 56% (1B) and 47% (2), and evaluated anamnestically. Statistical analysis was the analgesics were more cited, 84%, 94% and 100% performed by parametric (ANOVA) and non- to 1A, 1B and 2, respectively. For 1A, 45% reported parametric tests (U-Mann-Whitney test) and the the use of prescription drugs, of these critical level for rejection of the null hypothesis was antihypertensives (42%), hypolipidemics (14%) and taken to be a p value of 5%. Results: Our results proton pump inhibitors (11%). For 1B this frenquency revealed statistically significant elevations of the MDA was 56%, distribuited in antihypertensives (25%), and the OP concentrations in fungicides- exposed antidepressives (25%), hypolipidemics (11%) and workers versus controls (U-Mann-Whitney test and vitamins (11%). Among the non farmers the use was ANOVA, respectively; p < 0.05). There was no 21%, and the more frequent were antihypertensives significant difference between the exposed and (25%) and hypoglicemics agents (25%). Conclusion: controls comparing the MDA and OP concentrations Only thyroid disfuction was more reported among the and personal habits (smoking and drinking). farmers. The self-medication was similar between the Conclusions: The data obtained demonstrated that rural groups and the prescription drugs were more often workers exposed to fungicides presented excess of two reported among farmers, having a considered biomarkers, MDA and OP, which is important tools to percentual of antidepressive drugs. demonstrate body imbalance and, consequently, the installation of the oxidative stress process. Financial support / Acknowledgments: FUNDAÇÃO DE AMPARO À PESQUISA E Financial support / Acknowledgments: FAPEMIG INOVAÇÃO DO ESTADO DE SANTA CATARINA grant number [APQ-02281-16] (FAPESC); MINISTÉRIO DA SAÚDE, MS; CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E OC 10 - ENDPOINTS APPLIED TO A RISK TECNOLÓGICO, CNPq; SECRETARIA DE ASSESSMENT OF OCCUPATIONAL ESTADO DA SAÚDE, SES-SC; MINISTÉRIO PESTICIDE EXPOSURE PÚBLICO DO TRABALHO DE SANTA CATARINA (MPT-SC). MACHADO, S.C.1; SOUZA, B.M.1; PEREIRA, A.F.S.1; MARTINS, I.1.

OC 09 - OXIDATIVE STRESS AMONG Universidade Federal de Alfenas- MG. AGRICULTURAL WORKERS EXPOSED TO FUNGICIDES Introduction: Pesticides occupy a special position among various chemical agents that are present in the MACHADO, S.C.; MARCIANO, L.P.A.; ROSA, environment because of to their high toxicity, wide use D.R.S.; OLIVEIRA, C.S.; MORAES, G.O.I; and lack of action selectivity. Thus, it is essential that BRIGAGÃO, M.R.P.L; MARTINS, I. the risk assessment identifies safe levels of exposure. Universidade Federal de Alfenas-MG. This process is used to evaluate and organize data, information, estimates and uncertainties in a systematic fashion, with the purpose of understanding ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 187 and predicting the relationship between these exposure. To evaluate the human contamination by substances and their effects on the exposed non-target benzene, the measurement of urinary S- organisms. As a way of assessing adverse effects phenylmercapturic acid (SPMA) has been used. S- related to the use of pesticides, markers of exposure phenylmercapturic acid is a specific benzene called endpoints are used. Traditional endpoints often metabolite. Toxicokinetic studies are shown that only use highly predictive adverse effects or death; 0.11% of benzene entering the body is biotransformed however, there is a constant search for markers that in SPMA, with the elimination time equal to 9.6 hours. may determine the effects of exposure before the In this way, this metabolite has been indicated as a subjects suffer from them, and these markers are the specific biomarker of benzene exposure, even in long so-called surrogate endpoints. These surrogate working shifts. In order to evaluate the current endpoints are used to evaluate the incidence of situation of the petrol stations workers in Rio de genotoxic effects and can be used as biomarkers for Janeiro city, an integrative project is being developed assessing exposure to a particular pesticide or mixture. at the National School of Public Health Sergio Arouca Objective: This review discusses the importance of (ENSP). This project, entitled "Evaluation of these endpoints in risk assessments of occupational occupational exposure to benzene at gas stations in the pesticide rural exposure. Material and methods: city of Rio de Janeiro, consists in a cross-sectional Systematic review with qualitative data collection in study of the population of workers at gas station in several databases between November/2015 and comparison to an unexposed population. This project is December/2016. Results: Endpoints are still supported by the CESTEH / ENSP / FIOCRUZ, the established for a single pesticide. Thus, the use of Rio de Janeiro Municipal Health Secretariat (CEREST these is essential during all phases of risk analysis, and Environmental Surveillance) and from the first two stages when they are implemented SINDUSPETRO-RJ (Union of Employees in Fuel and until the risk characterization. At the end of the Oil Derivatives Services Offices of the State of Rio de process, the analysers must be able to estimate the Janeiro). Up to now, this study comprehends 51 risks due to exposure, indicate the general degree of occupationally exposed gas stations workers, mostly confidence in the estimated risks, cite the lines of those who directly handle the supply pumps, of 6 fuel evidence supporting the risk estimates, and interpret gas stations located of metropolitan region of Rio de the adverse effects. Conclusions: It is very important Janeiro and a control group (non-exposed workers), the knowledge and establishing endpoints to assess the consisting of 35 workers from the concierge and probability of an adverse effect occurring because of security service of the Oswaldo Cruz Foundation. The exposure to one or more pesticides aiming the validation of an HPLC analytical method is in course. healthcare of rural workers. The linearity and the matrix effect were already studied, and the obtained results showed no important Financial support / Acknowledgments: CAPES and interferences. FAPEMIG (APQ-02281-16) Financial support / Acknowledgments: CESTEH/ENSP/FIOCRUZ OC 11 - USE OF URINARY S- PHENYLMERCAPTURIC ACID IN THE EVALUATION OF BENZENE EXPOSURE TO OC 12 - OPEX CALCULATOR - SUPPORT FUEL STATIONS WORKERS IN RIO DE TOOL FOR OCCUPATIONAL RISK JANEIRO CITY ASSESSMENT

ROSA, J. C. S.; MOREIRA, J.C.1; FIGUEIREDO, AGUIRRE A.C; CAZARIN K.C.C; DIAS F.S; V.O.1; GONÇALVES, E.S.1; LARENTIS, A.L.1; GRIGOLI M.B; PALMA F.C; SARTI A. COSTA-AMARAL, I.C.1; CARVALHO, L.V.B.1; BORGES, R.M.1. Instituto ProHuma de Estudos Científicos

1 Centro de Estudos em Saúde do Trabalhador e Over the last 25 years, Regulatory Agencies have been Ecologia Humana/Escola Nacional de Saúde active on improving methodologies and data Pública/FIOCRUZ. generation to assess the exposure of rural workers to pesticides. Currently, the most used approach to The environmental and occupational exposure of evaluate their safety is the Occupational Risk workers at fuel stations to toxic chemical compounds Assessment. In Brazil, the risk assessment was cited in present in gasoline and other fuels are of public health Decree No. 4,074/2002, Art. 95, item III, which concern. Among those chemicals, benzene is the one regulates Law No. 7,802/1989. When considering the of greatest importance since it is a carcinogenic agricultural scenarios, it is known that workers can be substance. So, there is no safe limit to benzene exposed by the dermal and inhalation routes. In order ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 188 to evaluate the different agricultural scenarios, several adopted occupational safety and health (OSH) studies were carried out to monitor worker exposure practices and quantifying cholinesterase blood activity. and their results are used to comprise a generic A total of 65 employees was surveyed; most of them database. These data allow to extrapolate the exposure (78.5%) declared to perform a wide range of activities unit to other agrochemicals where the same activity is such as pruning, weeding, thinning and harvesting. performed (Popendorf et al., 1980 and 1982, Zweig et Despite that, they never received training for safe al 1984 & 1985). The aim of this work was to develop handling pesticides, nor had seen pesticide labels. a tool, called the OPEX Calculator - Support Tool for About 50% of workers did not use personal protective Occupational Risk Assessment that gathers equipment as recommended. Despite employees information of generic data of worker exposure when (63.1%) stated that restricted entry intervals were handling pesticides. This calculator will provide respected, such interval was associated with odor support for the evaluation of the occupational risk, as it dissipation. In general, a discrepancy between what the will allow estimating the exposure of individuals who workers said to do and to know and what they really handle agrochemicals on the field using different did and knew could be observed. A total of 50 workers application equipment”. The calculator provides have blood samples collected and analyzed. generic data - inhalation and dermal exposure unit - for Considering all subjects, significant activity inhibition different scenarios of occupational exposure that can was observed to acetylcholinesterase [3.64 U/ml (0.48) be assessed. It is a tool that allows the selection of vs. 3.38 U/ml (0.58); p=0.004], but not to specific data related to the conditions of use of the butyrylcholinesterase [1.62 U/ml (0.42) vs. 1.56 U/ml product and the scenario under evaluation, such as: the (0.42); p=0.194]. Also, significant acetylcholinesterase amount of active ingredient (AI) handled, the size of activity inhibition was observed for those handling the treated area, selection of the agricultural crop, flowers [3.62 U/ml (0.50) vs. 3.31 U/ml (0.59); specific toxicological data of the active p=0.003] but not for those responsible for pesticide ingredient/product under consideration as well as the spraying [3.70 U/ml (0.44) vs. 3.62 U/ml (0.50); personal protective equipment (PPE) that should be p=0.656]. These results suggest a lack of knowledge recommended to ensure the safety of the worker. With regarding OSH good practices and the adoption of a generic database and the insertion of specific usage practices that increase exposure. As a result, the parameters in the calculator it is possible to assess the potentially exposed population to anticholinesterase occupational risk for the selected scenario. The result pesticides is not exclusively those workers spraying is expressed as a percentage of the Acceptable pesticides, but all those workers continuously handling Operator Exposure Level (AOEL). the flowers.

Financial support / Acknowledgments: Instituto Financial support / Acknowledgments: Fundação ProHuma de Estudos Científicos. Jorge Duprat Figueiredo de Segurança e Medicina do Trabalho (Fundacentro)

OC 13 - OCCUPATIONAL EXPOSURE TO ANTICHOLINESTERASE PESTICIDES IN OC 14 - MAIN PESTICIDES USED BY WINE- FLOWER AND ORNAMENTAL PLANTS GROWERS IN SOUTHERN BRAZIL GREENHOUSES LINI, R.S.1; SILVA, L.F.F.2; AGUERA, R.G.3; NASSAR, P. P. M. 1; RIBEIRO, M. G. 1. OLIVERIA, N.G.1; NERY, B.G.1; SAKATA, A. S.1; SIQUEIRA, A.P.C.1; CAPELARI, S.4; 1 Fundação Jorge Duprat Figueiredo de Segurança e FROEMMING, E. O.1; NERILO, S.B.1,3; BANDO, Medicina do Trabalho (FUNDACENTRO), São Paulo E.1; OLIVEIRA, M.L.F.2; MOSSINI, S.A.G.1,5. - SP, Brasil. 1. Laboratory of Toxicology, Department of Basic Greenhouses are microcosms designed to optimize Health Sciences, State University of Maringá, Maringá plant growing and not to protect workers' health. – Paraná 2. Department of Nursing, Postgraduate Among rural workers, greenhouse floriculturists are Program in Nursing, State University of Maringá, suspected of being heavily exposed to agrochemicals Maringá – PR, Brazil 3. University Center Ingá, due to the enclosed environment and to the continuous Maringá – PR, Brazil 4. Paraná Institute of Technical contact with the plants. Cholinesterase inhibitors Assistance and Rural Extension – EMATER, agrochemicals are largely used by flower growers in Marialva-PR, Brazil 5. Postgraduate Program on Alto Tietê, an important region for the production of Bioscience and Physiopathology, State University of flowers and ornamental plants in greenhouses. This Maringá, Maringá – PR, Brazil. study aimed at investigating possible occupational exposure to anticholinesterase pesticides in flower greenhouse workers from such region, considering the ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 189

Introduction: The illness related to production methods 1; SOARES, M. C. F. 1; BAISCH, P. R. M. 1; among wine-growers in Brazil it´s still poorly MUCCILLO-BAISCH, A. L. 1; SILVA-JÚNIOR, F. documented in the literature. However, it is known that M. R. 1. Brazil is the largest consumer of pesticides in the world and Paraná is the third largest consumer in the 1 Universidade Federal do Rio Grande, Rio Grande - country. The massive use of pesticides brings serious RS, Brasil problems for the health of workers and population, as well as causing damage to nature by the degradation of The present study assessed the DNA damage in non-renewable natural resources, imbalance and environmentally exposed volunteers living in seven destruction of fauna and flora. Aims: Identify the main municipalities in an industrial coal region, through the classes of pesticides used by wine-growers in a city of use of the comet assay with blood cells and the Southern Brazil. Methods:A cross-sectional study was micronucleus test with buccal cells. The study aimed conducted in Marialva, Brazil with 190 wine-growers to conduct a cross-sectional study with men residing in exposed to pesticides. A Department of Health of the region of the coal industry to evaluate genotoxicity Paraná State guide named "Protocolo de Avaliação das and identify risk and protective factors associated with Intoxicações Crônicas por Agrotóxicos" was used for genotoxic outcomes. Blood and buccal smears were data collection. This study is approved by the Standing collected from 320 male volunteers living in seven Committee on Ethics in Research Involving Human cities inserted in a coal region. They were ages of 18 Subjects (COPEP) of the State University of and 50 years and also completed a questionnaire Maringa.Results: It was note a high use of pesticides in intended to identify factors associated with DNA viticulture, mostly fungicides it was verified the use of damage through a Poisson regression analysis. The 18 different substances belonging to the classes of comet assay detected significant differences in DNA fungicides, acaricides, bactericides and growth damage in volunteers from different municipalities, regulators. The most reported by the producers were and neighboring cities (Pedras Altas, Aceguá, and Dormex® and Cercobin® with 95.8%, followed by Hulha Negra) had a higher level of DNA damage in Cabrio top® (95.3%) and Score® (95.3%), being the relation to control city. Some of the risk factors fist a growth regulator classified as highly toxic, and associated with identified DNA lesions included the others fungicides of different toxicological classes. residence time and life habits. On the other hand, the The fungicides earn greater attention due to the high micronucleus test did not identify differences between prevalence and frequency of use, and the default of the cities studied, but the regression analysis identified legislation for this group. Fungicides dithiocarbamates risk factors such as age and life habits (consumption of as Cabrio top® provides severe headache, dizziness, mate tea and low carbohydrates diet). We conclude weakness, mental confusion, dyspnea, chest pain, that there are differences in the DNA damage of nausea, vomiting, abdominal pain, sweating, skin and volunteers from different cities of the carboniferous mucosal irritation. Despite the known clinical region, but the presence of micronuclei in the oral condition, in many cases the symptoms are trivialized mucosa does not differ between the same cities. by the workers, which make it difficult to diagnose Furthermore, we alert that some related factors may poisoning by pesticides. Conclusions: The results increase the risk of genotoxicity, such as residence indicate the need for intervention and treatment location and time, and living and food habits. Finally, strategies and disease prevention to the health of we suggest the need for continuous biomonitoring of workers and their families, as these products can cause the population, as well as for investing in health damage to workers' health when they are not used promotion in these vulnerable populations. safely. Financial support / Acknowledgments: CNPq Financial support / Acknowledgments: Acknowledgements: State Secretariat for Science, Technology and Higher Education –SETI; OC 16 - GENOTOXICITY IN BRAZILIAN COAL Management Unit of the Paraná Fund - SETI/UGF and MINERS AND ITS ASSOCIATED FACTORS Extension Program "University Without Borders" – USF, CNPq and Araucaria Foundation for their TAVELLA, R. A. 1; FERNANDES, C. L. F. 1; support. SOARES, M. C. F. 1; ALMEIDA, K. A. A. 1; GARCIA, E. M. 1; PINTO, E. A. S. 1; MUCCILLO- BAISCH, A. L. 1; SILVA-JÚNIOR, F. M. R. 1. OC 15 - GENOTOXICITY IN ADULT RESIDENTS IN MINERAL COAL REGION - A 1 Universidade Federal do Rio Grande, Rio Grande - CROSS-SECTIONAL STUDY RS, Brasil.

TAVELLA, R. A. 1; PINTO, E. A. S. 1; GARCIA, E. Recent studies have demonstrated the strong M. 1; ALMEIDA, K. A. A. 1; FERNANDES, C. L. F. association between miners’ workplace in relation to ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 190 time and mode of contact with coal, as well as the due to problems such as non-use of personal protective intensity with which these factors cause DNA damage. equipment (PPE) and scarcity of health surveillance The present study assessed the DNA damage in miners systems. OBJECTIVE: This study aimed to evaluate of an open-cast coal mine in the Southern Brazil the living habits and working conditions in a group of through the comet assay in blood cells and farmers who live in the northwest of Rio Grande do micronucleus test in peripheral lymphocytes and Sul, southern Brazil. METHODOLOGY: The exfoliated buccal mucosa cells. The present study aims individuals were invited to participate in the study by to evaluate the potential genotoxic and associated the researcher, who explained the purpose of the study. factors among coal miners, divided by degree of Individuals who agreed to participate signed a detailed exposure. Blood and buccal smears were collected consent form. The questionnaire was applied by the from 158 workers, who actively participate in different researcher in the house of each farmer, with an activities in coal mining, and 38 individuals living in interview time of approximately 40 minutes. The the same city but do not have participation in coal questionnaire used in this study is based on the model mining activities (control group). The workers were recommended by the International Commission of divided in 3 different groups, according to the level of Protection against Environmental Mutations and contact with coal extraction. The participants answered Carcinogens (ICPEMC), which is composed of a questionnaire intended to identify factors associated questions about life, health and work habits. This study with DNA damage through Poisson regression. The was approved by the Research Ethics Committee of results regarding oral mucosa micronucleus test Feevale University under number showed a significant difference (p < 0.001) of the 60093316.7.0000.5348. RESULTS: The study workers group 1 and 2 in relation to the control group, population was composed of 56 farmers, all males. The where the group 1 has a higher degree of exposure to mean age of the participants was 45 years. Regarding coal than group 2. For the lymphocyte micronucleus the educational level of the farmers, the majority have test and comet assay there were no significant completed high school (35.7%). The individuals difference between the exposed groups and control reported not using PPE. Some farmers reported using group. Significant results from the brute analysis masks only during the mixing of the pesticides, not showed an association between the outcome and the using any protective equipment during the spraying of fact of living in the municipality of the mining the product. None of the farmers use the complete PPE company and the exposure to radiation in the last 12 (mask, gloves, boots, apron, long pants and cap). The months. On the other hand, the multivariate analysis most reported symptom by farmers after contact with showed an association of the tail moment with pesticides was headache. When asked about chronic radiation exposure in the last 12 months. Thus, the diseases, hypertension and thyroid dysfunctions were findings of this study reveal genotoxicity in oral the most commonly reported diseases. mucosa cells of workers exposed to coal and that CONCLUSION: It was noticed that the population of workers with higher degree of contact with coal have a this study is aware of the toxic effects of exposure to more pronounced response. It is also noticeable that pesticides, but they reported not using PPE because of important factors like housing conditions and clinical the discomfort caused to them, especially when the exams should be always considered in a weather is very hot. genotoxicological evaluation. Financial support / Acknowledgments: Feevale Financial support / Acknowledgments: CNPq

OC 18 - NEW GUIDELINES FOR THE OC 17 - PESTICIDE EXPOSURE AND HEALTH CRITERIA FOR THE EVALUATION AND CONDITIONS AMONG FARMERS IN TOXICOLOGICAL CLASSIFICATION OF SOUTHERN BRAZIL AGROCHEMICALS

TANANDRA BERNIERI; DABIANA RODRIGUES; CRUZ, T. M.; DANZE, A.P. 1; PAULA, G. 1; ISADORA RANDON BARBOSA; PATRICIA LOPES, K.M.T. 1; PINHEIRO, F. 1. GROLLI ARDENGHI; LUCIANO BASSO DA SILVA. 1 INTERTOX Ltda. São Paulo - SP, Brasil

1. Universidade Feevale, Novo Hamburgo-RS, Brasil. Introduction: The Agência Nacional de Vigilância Sanitária – ANVISA submitted in 2016 the Public INTRODUCTION: Rural workers represent a Consultations (CP) n°260, 261, 262 which respectively population that is highly exposed to pesticides and thus presented draft resolutions on the criteria and are susceptible to large risks of intoxication. In requirements for toxicological evaluation, the developing countries, health problems in farmers, guidelines related to the toxicological information for associated with the exposure to pesticides, are higher labels and package insert and the criteria for ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 191 classification and comparison of the toxic effects of exogenous intoxication. Objective: To evaluate the agrochemicals, components, and related wood exogenous poisoning caused by exposure to toxic preservatives in order to improve the current agents during labor activity in the city of Primavera do legislation, harmonizing the toxicological evaluation Leste – MT. Material and Methods: This is a cross- procedures, classification and labelling of products sectional epidemiological study, based on the with the criteria established by the Globally secondary data provided in SINAN, between 2007 and Harmonized System of Classification and Labeling of 2015. The profile of the victims were evaluated (age, Chemicals – GHS adopted internationally and sex and schooling level) and the cases (toxic agents established by United Nations (UN). Objective: To involved, circumstances, type of exposure) registered introduce and discuss the proposed guidelines on in occupational intoxication. Results: There were 76 harmonization of toxicological assessment and cases of exogenous intoxication related to labor classification criteria, the use of hazard communication activity. These represented 32.9% of the total elements on agrochemical labels and package insert. poisoning that occurred during the study period. Methods: Regulations and standards in force in Brazil Regarding the profile of the victims, 88.2% were male, were consulted and evaluated critically and also the 93.4% were in the age group of 20 to 59 years and recent proposals for Resolutions submitted by 31.6% had only elementary education incomplete. ANVISA and the GHS criteria. Results and discussion: Agrochemicals (70.1%) of agricultural, domestic, CPs n°261 and 262 incorporate some of GHS guidance public health and rodenticides were the main toxic to old rules established by ANVISA, but not all health agents involved. There were also cases with chemical hazards established by GHS. The classification criteria, products (6.5%), toxic plants (5.2%), veterinary pictograms, hazard statements and precautionary products (1.3%), medicines (1.3%) and cosmetics statements incorporated into and proposed in the CPs (1.3%) and in 14.3% cases of the records did not present some divergences not being fully harmonized include the toxic agent involved. The most frequent with the GHS. Since the CP n°260 brings new conditions for the intoxications were the accidental toxicological evaluation by using alternative methods mode (60.5%), habitual use (28.9%), environmental to replace animal tests and prioritization of (5.26%), food intake (5.26%) and in one case toxicological analysis for products of low toxicity. The management error. Most of the poisonings were public consultations received some contributions that classified as acute single exposure (59.21%) followed were discussed at a public hearing, but the draft by acute repeated exposure (14.47%), the other ones regulations have not yet been finalized and approved (26.32%) did not present information on the type of by the Collegiate Board. Conclusion: The new exposure. Of the total of 76 cases only one was issued guidelines bring an advance to the legislation of CAT (Labor Accident Communication), and this agrochemicals that today is outdated, but they present poisoning occurred accidentally with chemical. some divergences that can impact the communication Conclusion: The predominance of cases of and classification of hazards. occupational intoxication in the male gender and by pesticides was evident. Their knowledge will assist in Financial support / Acknowledgments: INTERTOX the elaboration of preventive policies and educational Ltda. São Paulo - SP, Brasil actions with a focus on the victims and more frequent toxic agents.

OC 19 - EXOGENOUS POISONINGS DUE TO Financial support / Acknowledgments: Federal LABOR ACTIVITY IN THE OF PRIMAVERA University of Mato Grosso, Institute of Biological DO LESTE – MT, BRAZIL Sciences and Health.;UNIC - Educacional, Primavera SANTANA, V. T. P. 1; SUCHARA, E. A. 2; do Leste – MT. PEREIRA JUNIOR, E. C. 3.

1. Mestre e Professor nos cursos de Biologia, Farmácia e Agronomia na UNIC – Educacional, Primavera do Leste – MT, Brasil; 2. Doutora e Professora nos Cursos de Farmácia e Biomedicina na Universidade Federal de Mato Grosso, Instituto de Ciências Biológicas e da Saúde, Barra do Garças – MT, Brasil; 3. Farmacêutico no Centro de Saúde São José, Rondonópolis – MT, Brasil.

Introduction: The inadequate occupational exposure of man to chemical agents, physical and biological can result in pathologies, even leading to cases of

OTHERS ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 193

OT 01 - MORTALITY DUE TO OT 02 - STUDIES OF OPHIDIAN ACCIDENTS UNINTENTIONAL POISONING IN BRAZIL: OCCURRING IN QUERÊNCIA-MT DATA FROM INFORMATION SYSTEM FOR NOTIFIABLE DISEASES DESSBESSEL, A. 1; SUCHARA, E. A. 1.

OLIVEIRA COSTA, A.1; ALONZO, H.G.A.2 1 Universidade Federal de Mato Grosso, Campus Araguaia - MT, Brasil. 1 Universidade Estadual de Campinas, Campinas-SP, Brasil; 2 Universidade Estadual de Campinas, Among the accidents with venomous animals, snake Campinas-SP, Brasil. notifications occur in greater numbers, mainly in the north and center-west country regions. These accidents Introduction: Mortality due to unintentional poisoning are more serious and have a high mortality rate, has been a growing public health problem. In 2012, an therefore representing an aggravating factor for the estimated more than 190.000 deaths worldwide were population health. The present study objective was to caused by unintentional poisonings. As part of evaluate the Snakebite envenomation cases occurred in Sustainable Development Goal (SDG) 3 - Health, the Querência, Mato Grosso. A descriptive, non- United Nations (UN) members countries, including probabilistic and cross-sectional epidemiological Brazil, undertook the commitments to reduce survey was carried out between 2007 and 2016. In the substantially by 2030 deaths and illness from studied period, 77 accidents cases involving venomous chemicals and air, water and soil pollution and animals were registered. The accidents incidence rate contamination. Measuring the mortality from with venomous animals was 56.2 ± 28.1. There were unintentional poisonings may provide an indication of accidents with men, adults (67.6%), brown ethnicity effectiveness of Brazil’s healthy system in order to (45.5%) and low schooling (70.2%) predominance. achieving the SDG health target Objective: To The majority of accidents occurred in the rural area describe the mortality caused by unintentional (97.5%) and in the rainy season (71.5%). The ophidian poisonings in Brazil for a period of 6 years. Methods: accidents, 87 % were caused by the genus Bothrops, This is a retrospective and descriptive study related to 7.8% by the genus Lachesis and 5.1% of the cases by unintentional poisoning cases reported by the the genus Crotalus. Therefore, the accidents reported Information System for Notifiable Diseases (SINAN) for snakes in the studied period were characterized by during the period of 2010 to 2015. Data were collected accidents caused by the genus Bothrops, of mild and from SINAN, processed and evaluated by using moderate severity. They occurred in greater numbers Microsoft Excel®. The variables included were sex, in the September to May months, which correspond to age, toxic agent and main circumstance. Results: the rainy season. Men with low schooling and During the study period 448.499 poisoning cases were residents or farm workers accounted for most of the reported to SINAN, of which 30.6% cases and 0.17% victims. Thus, prevention programs against these deaths were classified as unintentional. Among 787 accidents and educating the population about snake deaths cases men were the most frequent (60.7%) aged genera are necessary. KEYWORDS: Snakes, 20 to 39 years old (38.4%). Pesticides were the most Bothrops, Bothrops. lethal agents involved in the poisoning cases among Financial support/Acknowledgments: men (15.7%) and medicines were most frequent among UNIVERSIDADE FEDERAL DE MATO GROSSO. women (33.9%). The main circumstances leading to poisoning were accidental (58.8%), of with 12.1% were classified as occupational circumstance. However, of total the poisonings reported to SINAN, intention was not classified in 17.461 (3.9%) cases, which suggests that deaths from unintentional poisoning may be underestimated. Conclusion: The mortality due to unintentional poisoning occurred predominantly in men, through accidental circumstances, by using pesticides. SINAN is important to estimate the epidemiology of poisonings, although underestimate number of cases. Financial support/Acknowledgments: MEC/CAPES.

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 194

OT 03 - BIOANALYTICAL METHOD OT 04 - EVALUATION OF THE HEMOLYTIC VALIDATION OF A NEW HETEROCYCLIC N- ACTIVITY OF GRAVIOLA EXTRACT PHENYLPIPERAZINE DERIVATIVE (LQFM 05) BY UHPLC-MS IN RAT LIVER SAMPLES FOR REOLON, A. 1 2; INDRAS, D. M. 1,2; CAZ, P. 1,2; PHARMACOKINETIC STUDIES KOERICH, S. 1,2; MAREK, C. B. 2; ITINOSE, A. M. 3. RAMOS, A. C. M. 1; MENEGATTI, R. 1; CHAVES, A. R. 1; REZENDE, K. R. 1. 1 Master's program in Pharmaceutical Sciences of University of Western Paraná, 85819110, Cascavel- 1 Universidade Federal de Goiás, Goiânia - GO, PR, Brazil; 2. Laboratory of Cellular Toxicology, State Brasil. University of Western Paraná, 85819110, Cascavel- PR, Brazil; 3. Assistence Center in Toxicology LQFM05 is a potential antipsychotic prototype drug (CEATOX), Hospital University of Western Paraná, under pharmacokinetic investigation. In order to 85806470, Cascavel-PR, Brazil. achieve this goal, a bioanalytical validation was performed on rat liver using an LC-MS / MS. Introduction Annona muricata L. is a fruitful plant Chromatographic separation was performed on Waters with pleasant flavor and aroma, that besides its feed X-Terra RP18 column (3.0 x 50.0 mm, 3.5 μm) at features it has several uses in popular medicine due its 40°C, with a gradient mobile phase of acetonitrile and antidiarrheal, antiparasitic, antirheumatic, anti-spasms 10 mM ammonium acetate at 0.5 mL/min and injection properties and slimming. Efficacy and safety of volume of 15 μL. The quantification was performed by popular preparations have not been scientifically MRM mode with m/z 349,20→157, 07 for LQFM05 proven, so some toxicity tests, as hemolysis test, can and m/z 285,08→257,08 for diazepam (IS) on a be use to evaluate its the toxic effects through the MicrOTOF-Q III mass spectrometry, with electrospray damage in erythrocyte cell membrane. Objective To ionization (ESI) source in positive mode. Their evaluate the toxic effect of graviola extract through in retention times (tR) were 5.6 min and 6.3 min, vitro evaluation of erythrocytes’ hemolytic activity. respectively. Sample clean up procedure for liver Material and methods In the study, human homogenate supernatant were set by protein erythrocytes, collected with EDTA of a healthy precipitation method in acetonitrile. No interferent individual, were used. In tubes containing 500 µl of peaks was seen at tR of compounds of interest. The human erythrocytes in 2%, 500 µl, 100 µl and 10 µl of average amount of LQFM05 and IS recovered from graviola extract were added, completing with liver samples were consistent at all QC levels showing phosphate buffered saline (PBS) for a final volume of values (mean ± RSD) of 67.2% ± 10.7% and 82.4% ± 1 ml. In the negative control, suspensions of 8.5%, respectively. Calibration curves were plotted by erythrocytes and PBS were used, and, for the negative peak height ratio of drug/IS vs nominal drug control, a lysis buffer (2% acetic acid). In the concentrations. It showed to be linear in the range of sequence, the suspensions of erythrocytes in 2% were 10.0 to 900.0 ng/mL during both days of method incubated in 37ºC for 1 hour. After the incubation, the validation (r = 0.99994, y = 0.00170 x + 0.00796 and r cells were deposited in 377 g for 10 minutes. The = 0.99951, y = 0.00168 x + 0.00409; F-test: 0.47 and supernatant was collected (200 µl) and placed in 0.32, respectively). The intra- (n=6) and inter-day ELISA plate and the absorbance determined in 450 (n=12) precision ranged from 1.2 to 3.6% while the nm. The test was executed in triplicate and the results accuracy were within -3.6 to 7.5% and – 3.6 to 8.0%. were expressed in percentage of hemolysis. Results The matrix effect values of LQFM05 at three levels of The cytotoxic effects of the extract in human QC samples were in the range of 98.1 – 103.9%, and erythrocytes revealed hemolytic activity in the the RSD was ≤11.5%; the mean matrix effects of IS concentrations of 200 mg.ml-1 (38,04%) and 20 were 101.5%, the RSD of 4.9%. Stability data for mg.ml-1 (39,27%), with higher percentage of LQFM05/IS in the biological matrix after three freeze- hemolysis in the concentration of 20 mg.ml-1. In the thaw cycles (RSD ≤3.0%), after samples storage at - negative control, they didn’t have hemolysis after the 80°C for 1 week. Additionally, short-term (35 h/15°C), exposure to human erythrocytes, indicating that post-processing (-2 to 8°C; 58 h) and autosampler erythrocytes resist to the experimental conditions of (15°C; 24h) stability samples were also evaluated the test, while in the positive control, they had showing %RSD values lower than 3,3%. Taken all significant lysis, validating the test. Conclusion The together, the bioanalytical LC-MS method could be hemolysis test showed cytotoxic effect in the extract of validated for analysis and quantification of LQFM05 human erythrocytes. The high hemolytic activity in rat liver samples according to ICH guidelines. presented by graviola in the concentration of 20 Financial support/Acknowledgments: CNPq, Fapeg, mg.ml-1 may be justified by the presence of saponins CAPES, INCT/INOFAR, Funape, IQ/UFG, CPMG - that interact with the components of the erythrocyte Waldemar Mundim. cell membrane. ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 195

Financial support/Acknowledgments: Fundação Conclusion The hemolysis test showed cytotoxic effect Araucária, CAPES e CEATOX. in extract human erythrocytes extract. The high hemolytic activity presented by pink Ipê at 20 mg.ml-1 may be justified by the presence of alkaloids, steroids OT 05 - MUTAGENICITY OF AYAHUASCA and flavonoids, hence these substances have cytotoxic BEVERAGE TO THE SALMONELLA power. MICROSOME ASSAY USING THE OECD Financial support/Acknowledgments: Fundação RECOMMENDED STRAINS Araucária, CAPES, UNIOESTE e CEATOX.

REOLON, A. 1 2; INDRAS, D. M. 1 2; CAZ, P. 1 2; KOERICH, S. 1 2; MAREK, C. B. 2; ITINOSE, A. M. OT 06 - DETERMINATION OF ABIRATERONE 3 IN HUMAN PLASMA BY HPLC-FL

1 Master's program in Pharmaceutical Sciences of FURTADO, C.M.; QUADROS, N. D. 1; VERZA, S. University of Western Paraná, 85819110, Cascavel- G. 1. PR, Brazil; 2. Laboratory of Cellular Toxicology, State University of Western Paraná, 85819110, Cascavel- 1 Universidade Feevale, Novo Hamburgo - RS, Brasil. PR, Brazil; 3. Assistence Center in Toxicology (CEATOX), Hospital University of Western Paraná, Introduction: Prostate adenocarcinoma is the second 85806470, Cascavel-PR, Brazil. most common type of cancer in Brazil, with high rates in the South and Southeast regions. Among the drugs Introduction Tabebuia avellanedae Lorentz ex Griseb, available for the treatment of the disease, abiraterone popularly known as pink ipê, is frequently used in acetate is a new therapeutic option for the treatment of popular medicine due its anti-infective, antifungal, patients diagnosed with castration-resistant metastatic diuretic and astringent properties and for the treatment prostate cancer. Few methods are available to quantify of lupus, psoriasis and allergies. Pink ipê presents on abiraterone in human plasma using LC-MS/MS, its phytochemical analysis, secondary metabolites however only one method uses HPLC-FL. Objectives: including saponins, flavonoids, coumarins, tannins and Considering this the objective of this study was to lapachol. The efficacy and safety of popular develop and validate an analytical method for preparations have not been scientifically proven, so determination of abiraterone in human plasma using some toxicity tests can be used such as the hemolysis HPLC-FL. Methodology: To extract abiraterone of test that evaluates the toxic effects of a substance human plasma, liquid-liquid extraction with a mixture through damage in erythrocyte cell membrane. of n-hexane:isopropanol (85:15 v/v) was used. Objective Evaluate the erythrocytes’ hemolytic activity Chromatographic analyses were performed with a of pink ipê extract, to analyze its possible toxic effects. Shimadzu Class VP high performance liquid Material and methods In the study, were used human chromatograph and a Thermo C18 (150 x 4,0 mm, 5,0 erythrocytes collected with EDTA. In tubes containing um) column, maintained at 25°C. The mobile phase 500 µl of 2% human erythrocytes, were added 500 µl, consisted of potassium phosphate buffer 25mM (pH 100 µl and 10 µl of pink ipê extract completing with 5.8) and acetonitrile (30:70 v/v) in isocratic mode. A phosphate buffered saline (PBS) for a final volume of variation in flow rate was applied: 0.1-7.0 min.: 1,0 1 ml. For negative control, suspensions of erythrocytes mL/min.; 7.5 min.-20 min.: 1,3 mL/min.; 20-23 min.: and PBS were used, and for the positive control lysis 1,0 mL/min. Chromatograms were acquired at an buffer (2% acetic acid) were used. In sequence, the excitation wavelength 255 nm and an emission suspensions of 2% erythrocytes were incubated in wavelength 340 nm. Detector sensitivity was set at 37ºC for 1 hour and after this, the cells were deposited high sensitivity and gain of 1. The method was in 377 g for 10 minutes. The supernatant was collected validated following FDA US guidelines for (200 µl) and placed in ELISA plate and the absorbance bioanalytical method validation. Results: Total determined in 450 nm. The test was made in triplicate chromatography running time was 22 min. Inter-day and the results were expressed in percentage of precision showed CV% was below 5,19 % and intra- hemolysis. Results The extract cytotoxic effects in day precision showed CV% ≤ 7.78 %. Accuracy was human erythrocytes revealed hemolytic activity at 200 between 97.49 – 99.53 %. The lower limit of mg.ml-1 (47,42%) and 20 mg.ml-1 (52,83%) quantification was 5 ng/mL. Mean extraction yield of concentrations, with higher hemolysis percentage at 20 abiraterone was 84.04%. During stability assays the mg.ml-1. The negative control didn’t showed abiraterone was stable in plasma at room temperature hemolysis after human erythrocytes exposition, for 12 h and it was also stable after three freeze-thaw indicating that erythrocytes are resistant to cycles at -20ºC. Conclusion: A sensitive method for experimental test conditions, while in the positive determination of abiraterone in human plasma was control, they had significant lysis, validating the test. developed and validated and can be applied to drug ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 196 therapeutic monitoring of patients in treatment with and to environmental exposures present in rural life abiraterone acetate. that may be related to PD. Financial support/Acknowledgments: CAPES/ Financial support/Acknowledgments: CNPq Universidade Feevale. (134411/2015-3); Núcleo de Avaliação do Impacto Ambiental sobre a Saúde Humana (TOXICAM).

OT 07 - PARAQUAT AND PARKINSON’S DISEASE: A SYSTEMATIC REVIEW AND OT 08 - PARTICULARITIES OF EXOGENOUS META-ANALYSIS OF OBSERVATIONAL INTOXICATION IN ADULTS AND ELDERLY STUDIES. IN JATAÍ, GOIÁS.

VACCARI, C. 1; EL DIB R. 2,3; DE CAMARGO, SANTOS, K. F. 1; OLIVEIRA, F. F. S. 1; SUCHARA, J.L.V. 1 E. A. 1.

1 Universidade Estadual Paulista (UNESP), Faculdade 1 Universidade Federal de Mato Grosso, Campus do de Medicina de Botucatu, Botucatu - SP, Brasil;2 Araguaia, Barra do Garças - MT, Brasil. Universidade Estadual Paulista (UNESP), Instituto de Ciências e Tecnologia, São José dos Campos - SP, Introduction: Investigations of particularities of Brasil; 3 Universidade McMaster, Instituto de intoxications in elderly people are necessary, regarding Urologia, Hamilton - ON, Canadá. that aging is a worldwide tendency and they face a natural physiologic decline. Objective: To characterize Introduction: Parkinson's disease (PD) is a progressive the epidemiology of exogenous intoxications among neurodegenerative condition that has aging, genetic adults and elderly population in Jataí, Goiás. Materials susceptibility and exposure to certain chemicals as risk and Methods: This is a descriptive cross-sectional factors. In the last decades, epidemiological and epidemiologic study referred from 2009 to 2014. Data experimental studies have suggested an association were processed using Microsoft Excel, and evaluated between the herbicide paraquat and the development of through the BIOESTAT statistical software. The age PD. Objectives: To verify an association between range was established according to the Brazilian paraquat and PD through systematic review of the Institute of Geography and Statistics, comprising literature and meta-analysis. Methods: Observational adults aged from 20 to 59 years old and elderly aged studies available in the PubMed, EMBASE, LILACS, 60 years and older. Results: In this period, it was TOXNET and Web of Science databases were selected observed 640 adult intoxications (93,2%), emphasizing without language or publication period restrictions. the feminine gender (54,5%) affected by drug The heterogeneity among studies was evaluated by the intoxication (43%), the main circumstance the suicide I² test. Dichotomous data were summarized as odds attempt (56,6%). There were 47 elderly intoxications ratios with their respective 95% confidence intervals (6,8%), predominated by masculine gender (51,1%), and risk of bias was assessed by modified tools from which the main toxic agent was pesticides (38,3%), the NewCastle Otawa Scale. Results: Fifteen studies accidentally occurred (42,5%) and presenting higher published in English in the period 1994-2014 were lethality (6,4%). The lethality was five times higher included, one being a cohort study, 13 case-control than in adults (1,2%). The mean rate of suicide attempt studies and one cross-sectional study. The main biases was 7 times higher in adults (134,3/100.000 hab.) than identified were related to exposure assessment, in the elderly (18,7/100.000 hab.). Conclusion: It is selection of controls and adjustment for confounding concluded that the studied groups have heterogeneous factors. A significant OR of 1.40 (95% CI: 1.16-1.70, intoxication profiles and then, the health organizations I² 15%) was obtained, but the majority of the studies have to propose educational campaigns and treatments had a very small number of participants exposed to specifically directed to each group. paraquat and were at high risk of bias. Applying Hill’s Financial support/Acknowledgments: Secretaria de “criteria” it was not possible to affirm that the Saúde e a Vigilância Epidemiológica do município de association between paraquat exposure and PD was of Jataí, GO. causality. Conclusion: In order to confirm the small positive association between paraquat and PD, the need remains for new prospective studies, with careful assessment of paraquat exposure, preferentially through determination of herbicide residues in urine or blood samples of potentially exposed subjects, and more participants exposed to the risk factor. It is also important to evaluate mutations in genes that increase susceptibility to PD, the coexposure to other pesticides ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 197

OT 09 - PROFILE OF PATIENTS OT 10 - QUALITY ASSESSMENT OF DATA INTOXICATED BY MEDICINES FROM POISONING WITH MEDICINES INVESTIGATED BY TIC-GO FROM 2007 TO REGISTERED IN SINAVISA 2012 OLIVEIRA, F. N. M. 1; CUNHA, L. C. C. 1; ZATTA, OLIVEIRA, F. N. M. 1; CUNHA, L. C. C. 1; ZATTA, D. T. 2; LINS, J. B. A. 2; BARBOSA, M. A. 1 D. T. 2; BARBOSA, M. A. 1. 1 Universidade Federal de Goiás - Goiânia-GO; 2 1 Universidade Federal de Goiás - Goiânia-GO; 2 Secretaria de Saúde do Estado de Goiás. Secretaria de Saúde do Estado de Goiás - Goiânia – GO. Background: The record of intoxication data is one of the responsibilities of the Toxicological Information Background: The large necessity to improve the study Centers (TICs).The criteria standardization that of public toxicology data in Brazil, especially involves the health problems classification favors the medicines. Methodology: It was performed a statistical registration of epidemiological data and, consequently, analysis of 5041 sheet of research from SINAVISA the possibility of comparing the records data between system of patients who were drug intoxicated through different regions as well as over time. Goals: Assess Multiple Correspondence Analysis and Hierarchical the quality of the data registration of variables: product Cluster Analysis in order to get the profile of medicine and "chemical group" in 5041 investigation files that intoxication cases of patients investigated by were registered from 2007 to 2012 in TIC from Goiás Toxicological Information Center of the State of Goiás state. Methodology: The nominal data referring to (TIC-GO) from 2007 to 2012 and verify the probable the108 "chemical groups" of 609 products registers in identity of the unknown products used by patients in SINAVISA were submitted to Textual Analysis intoxications. Results: 20.49% of information was through SPAD Statistical Program. Results: In the obtained in the first two axes of the data matrix textual analysis of variable: product and "chemical represented in a contingency table. The first Multiple group", it was observed the grouping of similar ratings, Correspondence factorial axis separated the anti- making it possible to verify incorrect classifications. depressant and anti-depressant products from anti- There were mistakes that were probably made by inflammatories, antihistamines and adrenergic professionals who are not from health field. Of the 609 agonists, while the second factorial axis separated products and 85 "chemical groups" 231 were patients under 14 from those older than 20 years. In the reclassified and 86 presented Portuguese errors. Hierarchical Cluster Analysis, the data were allocated Conclusion: The proposed method was able to in fifteen groups (clusters). Conclusion: It is concluded demonstrate the errors in the data collected. A that the profile of intoxicates are represented by correction list was generated for the classification of patients of the female gender and from 1 to 4 years and products and "chemical group" that currently the TIC- 20 to 29 years. Patients from 1 to 4 years intoxicate GO is following by default. with antibiotics, no steroidal anti-inflammatory drugs, Financial support/Acknowledgments: Universidade antipsychotics, adrenergic agonists, hormones and Federal de Goiás. antihistamines, and patients from 20 to 29 years old intoxicate with anxiolytics, antidepressants and anticonvulsant. The 30 ignored products in cluster IX OT 11 - ENDOCRINE DISRUPTORS AND may be anxiolytic or antibacterial, the 23 ignored BRAIN SEXUAL DIFFERENTIATION "chemical groups" in cluster XIII may be anxiolytics, anticonvulsants, antidepressants or antipsychotics, and DA SILVA, G. N. 1; DALSENTER, R. P. 1; the 300 ignored products in cluster XI are most likely MENEGATI, S. E. L. T. 1 antibacterial. Financial support/Acknowledgments: Universidade 1 Universidade Federal do Paraná, Curitiba - PR, Federal de Goiás e Secretaria da Saúde do Estado de Brasil. Goiás. Introduction: The differences between genders are quite clear, mostly in sexual behavior. This occurs due to sexually dimorphism in some brain areas, mainly the hypothalamus, which is responsible to regulate some behaviors. Furthermore it is known that these differences are determined by intrauterine hormonal environment which differs from male and female fetuses. The hormones act in brain development and masculinize or feminize areas that contain steroidal ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 198 receptors. Moreover, there are chemicals called which had the uterine horn exposed in vitro to endocrine disruptors (ED) that act like hormones. betamethasone on gestational day 19; BM (n=5), Objective: The aim of this study was to describe treated with 0.1mg/Kg/day of betamethasone; and sexually dimorphic brain regions and evaluate if Control (n=5), treated with saline; BM and Control intrauterine exposure to ED can affect dimorphic groups were treated with na intramuscular injection on structures of the brain. Material and Methods: gestational days 12, 13, 18 and 19. In BMiv group we Biomedical literatures and database were consulted to evaluated the contractility of the uterus associated with do this review using the key words gestation, lactation, acetylcholine. In BM and Control groups we evaluated endocrine disruptors, brain sexual differentiation. the effects of in utero exposure to betamethasone on Results: The brain sexual differences are well male offspring, by the body and organ weights, established in animal models. In humans, it does not anogenital distance (AGD) at postnatal day (PND) 1, occur due to the difficulty in evaluate the effect of and histomorphometric analysis of the testicles at PND different situations in the human brain. However, it is 7, 14 and 28. Results: In vitro exposure to known that the sexually dimorphic nucleus of pre optic betamethasone decreased the contractility of uterine area and amygdala are dimorphic and some studies tissue. In utero exposure to betamethasone decreased show the relationship between the changes in these the body weight and AGD of male offspring at PND 1. brains areas, relating them with the sexual orientation Furthermore, at PND 14 and 28, the animals of BM and gender identity. Besides that, some papers indicate group presented lower body weight. At PND 14, that ED can change sexually dimorphic brain areas of testicles and epididymides weights were also reduced. rodents. These changes lead to difference in sexual Histomophometric analisys revealed a decrease in differentiation and to a variation in sexual behaviors. Leydig cells nuclear volume at PND 7, 14 and 28, and In humans, intrauterine exposure to ED was related to lower number of Sertoli cells at PND 14 and 28, in BM cryptorchidism and hypospadias. Conclusion: This group. Conclusion: Our results suggest that gestational review showed that some dimorphic brain structures exposure to betamethasone potentially impairs are responsible for the sexual behavior and some ED pregnancy and postnatal reproductive development. could modify structures resulting in different sexual This data raise concerns about the use of behaviors. betamethasone on human antenatal therapy. Financial support/Acknowledgments: CAPES, Financialsupport/Acknowledgments: FAPESP grant CNPq. #2017/02764-9.

OT 12 - GESTATIONAL EXPOSURE TO OT 13 - BDE-153 CAUSES TOXIC EFFECTS TO BETAMETHASONE IN RATS IMPAIRS HepaRG CELL LINE LEADING TO CONTRACTILITY OF GRAVIDIC UTERUS MITOCHONDRIAL AND CELLULAR AND POSTNATAL REPRODUCTIVE DAMAGE. DEVELOPMENT OF MALE OFFSPRING TASSO, M.J. 1; MIRANDA, R. G. 1; PEREIRA, L. C. BARROS, J.W.F. 1; BORGES, C.S. 1; FIGUEIREDO, 2; DORTA, D.J. 1 T.M. 1; PACHECO, T.L. 1; ROSA, J.L. 1; LOZANO, A.F.Q. 1; SILVA, K.P. 2; PUPO, A.S. 2; KEMPINAS, 1 Departamento de Química, Faculdade de Filosofia W.G. 1 Ciências e Letras de Ribeirão Preto, Universidade de São Paulo-USP, Ribeirão Preto - SP, Brasil; 2 São Paulo State University (UNESP), Institute of Departamento de Bioprocessos e Biotecnologia, Biosciences, Botucatu, São Paulo – Brazil, 1. Faculdade de Ciências Agronômicas de Botucatu, Department of Morphology; 2. Department of Universidade do Estado de São Paulo-UNESP, Pharmacology. Botucatu-SP, Brasil.

Introduction: Preterm birth risk require rapid several PBDEs (Polybrominated diphenyl ethers) belong to a maternal and fetal physiological modifications. class of flame retardants that is commonly used in Therapy with betamethasone, a synthetic manufactured products. The BDE-153 congener had glucocorticoid used in this case promotes fetal lung already shown evidences of toxicity, which combined maturation, which decreases neonatal mortality and with its high bioaccumulation capacity, and its morbidity. Objective: Thus, the present study aimed to increasing levels on biotics and abiotics systems, has evaluate the impact of betamethasone on gravidic brought growing concerns about its presence in the uterus contractility and postnatal reproductive environment and the consequences to the human development of male offspring exposed in utero to this health. Therefore, the aim of this work was to synthetic glucocorticoid. Methods: Pregnant Wistar investigate the toxic effects of BDE-153 using the rats were allocated into three groups: BMiv (n=5), HepaRG cells line (derived from a human ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 199 hepatoblastoma, and with a great similarity with the literature. Kaempferol and morin are two flavonoids metabolic system of a human healthy liver) as which possess structures very similar to each other, experimental model to clarify the risk of BDE-153 differing only in the presence of an extra 2'-hydroxyl exposure. Briefly, cytotoxic effect of BDE-153 was substituent (B-ring) in morin. This study was aimed to investigated at concentrations ranging from 0.1 to 25 assess the effects of kaempferol and morin, at µM, with 24h and 48h exposure. Cell viability by the concentrations ranging from 1 µM to 100 µM, in MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl HepG2 cells to check for induction of genotoxicity and tetrazolium bromide) assay and cell proliferation using cytotoxicity, and to verify if the structural difference the dye sulphorhodamine B were assessed. The BDE influences the toxic effects. The HepG2 cell line was influence on the generation and accumulation of exposed to each flavonoid singly and submitted to reactive oxygen species (ROS), and in the maintenance assays to examine proliferation, viability, of mitochondrial membrane potential was also mitochondrial membrane potential maintenance, measured using the fluorescent probes CM-H2DCFDA accumulation of reactive oxygen species, (5,6-chloromethyl-2′,7′-dichlorodihydrofluorescein phosphatidylserine exposure in the outer membrane, diacetate acetyl ester), and TMRM nuclear fragmentation, release of caspases, cell cycle, (tetramethylrhodamine methyl ester), respectively. We extracellular lactate dehydrogenase activity, and observed the ability of BDE-153 to inhibit cell genotoxic effects. Kaempferol, at 50 and 100 µM, proliferation and decrease cell viability in reduced cell proliferation and viability, interfered with concentrations of 10 μM and higher. BDE-153 had the mitochondrial electrochemical potential, did not also caused a decrease in mitochondrial membrane provoke a significant accumulation of reactive species, potential in concentrations of 10 and 25 μM already and demonstrated to cause cell death by apoptotic after 24 hours of treatment, thus demonstrating pathway, evidenced by the raise in exposure of evidences of damages on mitochondria functions that phosphatidylserine on the outer cell membrane, the can lead to cell death. In addition, it was observed a increased nuclear fragmentation, and ability to activate significant increase in reactive oxygen species caspases -3 and -9. Kaempferol, also caused cell cycle accumulation, which may be responsible for the arrest at the G2 phase, induced LDH release, and mitochondrial damages. Thus, our results suggest that displayed genotoxic effects. On the other hand, at BDE-153 causes cytotoxic effects in HepaRG cell, concentrations tested, morin did not affect cell perhaps leading to cell death related to a mitochondrial proliferation and viability, did not increase the amount dysfunction. of reactive species, nor showed evidences of apoptotic Financial support/Acknowledgments: FAPESP e process, causing only a decrease in mitochondrial CNPq. membrane potential, and inducing genotoxic damage in higher concentrations (50 e 100 µM). Thus, it was possible to verify that the structural differences OT 14 - AN EXTRA HYDROXYL-GROUP IN between the flavonoids tested were sufficient to enable THE B-RING OF FLAVONOID MORIN the occurrence of clearly distinct effects between this PROVIDES DISTINCT TOXIC EFFECTS IN two flavonoids (kaempferol and morin), which may be HEPG2 CELLS, COMPARED TO FLAVONOID attributed to the extra hydroxyl group in the morin KAEMPFEROL. structure. Financial support/Acknowledgments: CNPq e TASSO, M.J. 1; OLIVEIRA, A. S. 1; BERNARDES, FAPESP. M. F. F. 2; PEREIRA, L. C. 3; DORTA, D.J. 1.

1 Departamento de Química, Faculdade de Filosofia Ciências e Letras de Ribeirão Preto, Universidade de São Paulo-USP, Ribeirão Preto - SP, Brasil; 2 Departamento de Análises Clínicas Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo-USP, Ribeirão Preto - SP, Brasil; 3 Departamento de Bioprocessos e Biotecnologia, Faculdade de Ciências Agronômicas de Botucatu, Universidade do Estado de São Paulo-UNESP, Botucatu - SP, Brasil.

Flavonoids are polyphenolic compounds which have antioxidant properties, and whose mechanisms of action are influenced by their structural characteristics. However, toxic effects have also been shown in ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 200

OT 15 - ANTIMICROBIAL SUSCEPTIBILITY present at 200 mg.mL-1 A. muricata extract TEST WITH ESCHERICHIA COLI STRAINS IN concentration. This result is according the literature. ANNONA MURICATA EXTRACT Financial support/Acknowledgments: Unioeste; CEATOX; CAPES; Fundação Araucária. CAZ, P.1,2; REOLON, A.1,2; INDRAS, D. M.1,2; KOERICH, S. 1,2; MAREK, C. B2; ITINOSE, A. M.3. OT 16 - REGULATORY REQUIREMENTS FOR GENOTOXICITY ASSESSMENT 1.Master´s Program in Pharmaceutical Sciences of Western Paraná State University, Cascavel - PR, BOOTH, E.D 1; RAWLINSON, P. J. 1; FAGUNDES, Brazil. 2.Laboratory of Cellular Toxicology, Western P. M. 2; LEINER, K. A. 3 Paraná State University, Cascavel- PR, Brazil. 3.Toxicology Assistence Center (CEATOX), Hospital 1Department of Toxicology and Health Sciences, of Western Paraná State University, Cascavel-PR, Syngenta Ltd, Jealott’s Hill; 2Department of Product Brazil. Safety, Syngenta Protecao de Cultivos Ltda, Sao Paulo; 3Department of Toxicology and Health Introduction: The emergence of microbial resistance Sciences, Syngenta Crop Protection. tends to increase with the misuse of antibiotics, Escherichia coli is a micro-organism with a great Introduction: Plant protection products (PPP) have importance in epidemiology scenario. Annona been developed to reduce crop losses and maintain muricata, popularly known as graviola, is a plant that yield for farmers and growers. As a core element of has many popular uses: boiled leaves are used to their safe use, development, and governance, the combat diarrhea, spasm, as slimming and against human safety of the active ingredients is addressed cancer. Its shell has antidiabetic and spasmolytic across a range of toxicology end-points with an action. It presents metabolites with microbial and assessment of genotoxicity of the active ingredient modulatory activity such as tannins, flavonoids and being a key component. Objectives The objective of flavones. The presence of annonacina results in this is to review the current, and in some cases toxicity on neurons and suggests that this substance is proposed, genotoxicity data requirements in different the probable cause of the onset of parkinsonian global territories for chemical active ingredients as syndrome. In Brazil, popular preparations containing well as biologicals, microbial, ground water A. muricata are common but its efficacy and safety metabolites, metabolites, and impurities. Material and have not been scientifically proven, despite sheets, methods: The method employed included searching for they are considered potentially toxic to humans. requirements regarding genotoxicity data in the main Objective: Evaluate the antimicrobial activity of A. regulatory agencies, considering their different muricata extract at 2 mg.mL-1, 20 mg.mL-1 and legislations, as well as the test guidelines available for 200mg.mL-1 concentrations by E. coli strain. Material the standardized generation of genotoxicity test data. A and methods: The microbial inoculum was prepared total of 52 regulatory requirements documents were with E. coli ATCC 25922 standard strain (of 24 hours analysed. Results: There are no overarching global incubation) and suspended in sterile saline solution. agreements on which genotoxicity studies need to be The filter paper disks were impregnated with 20uL of conducted to satisfy the majority of regulatory 2 mg.mL-1, 20 mg.mL-1 and 200mg.mL-1 A. muricata authorities. The implementation of new OECD extract concentration. As negative control ethanol 4.50 guidelines for the in vitro micronucleus, transgenic mg.mL -1 and distilled water were used and 30 μg rodent somatic and germ cell gene mutation and in ceftazidime disk was used as positive control. The vivo comet assays, as well as the revision of some disks were kept in an oven for 10 minutes at 37°C and other OECD test guidelines has resulted in some then were distributed over the surface of the culture changes to data requirements. Discussion and medium, previously seeded with bacterial inoculum. conclusion: Based on the results, it is possible to The plate was incubated inverted at 37° C for 24 hours. conclude that regarding genotoxicity data requirements Was considered active the samples that presented between countries or regions, there are greater growth inhibition halo greater than or equal to 8 mm similarities than differences. Also, the tiered in vitro diameter. Results: The 200 mg.mL-1 sample showed and in vivo testing strategies have been shown to be active in front of the tested strain, presenting average protective of human health, which consequently leads of 12mm inhibition halos. 20 mg.mL-1 and 2 mg.mL-1 to a more reticent position on adopting new test showed no activity. The positive control had a mean of methods. However, current developments in this area 28mm halos and the negative controls presented 0mm show a keen awareness predicated on the in both samples. Conclusion: The antimicrobial demonstration of the safe use of PPPs that are highly activity front negative Gram bacterium E. coli was beneficial.

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OT 18 - EVALUATION OF THE TOXICITY OF OT 17 - EFFECT OF SOLUBILITY ENHANCERS UNHA DE GATO (Uncaria tomentosa) ON ON RAT INTESTINAL TISSUE VIABILITY AND Artemia salina AQUEOUS SOLUBILITY OF BROSIMUM GAUDICHAUDII TRÉCUL (MORACEAE) KOERICH, S.1,2; INDRAS, D. M.1,2; REOLON, EXTRACTS A.1,2; CAZ, P.1,2; MAREK, C. B.2; ITINOSE, A. M.3 MACHADO, R.D.1; CAPUZZO, I.M.C 1; COSTA, E. T. S. 1; REZENDE, K. R. 1 1Master's program in Pharmaceutical Sciences of University of Western Paraná, 85819110, Cascavel- 1 Universidade Federal de Goiás, Goiânia – GO, PR, Brazil; 2Laboratory of Cellular Toxicology, State Brasil. University of Western Paraná, 85819110, Cascavel- PR, Brazil; 3Assistance Center in Toxicology Introduction: Psoralen (Pn) and bergapten (Bn) are the (CEATOX), Hospital University of Western Paraná, major low solubility furanocoumarins compounds 85806470, Cascavel-PR, Brazil. from Brosimum gaudichaudii Trécul (EBGT). Solubility enhancers are often used to assess intestinal INTRODUCTION: Medicinal plants are largely used permeability of poor water-soluble drugs. Thus, in traditional practices and basic health care. Uncaria evaluation of its impact on reducing tight junctions tomentosa, popularly known as unha de gato, is a integrity and rat jejunal tissue viability by means of bindweed woody plant that is used as therapy in the transepithelial electrical resistance (TEER) is crucial. treatment of asthma, arthritis and rheumatism. Toxicity Thus, the aim of this work was to assess some tests are elaborated to assess and predict the substances solubility enhancer´s impact over TEER measurements toxic effects on biological systems. Bioassays tests are and also, the aqueous solubility of EBGT, expressed as used to evaluate plants toxicity, including Artemia Pn and Bn concentrations. Materials and methods: salina that is a saltwater microcustacean. Simplicity, Solubility enhancers were based on co-solventes (5 quickness and low cost favor its use in preliminary and 10% PEG 400; 20% PG; 5% Tween 80; and 0.1 analyses of general toxicity. OBJECTIVE: The present and 0.5% DMSO) and complexation agents (HP-β-CD study verified the mortality percentage of Artemia 5, 10 and 20% w/w) were evaluated according to salina in different concentrations of unha de gato shake-flask modified method. Markers content were extracts. MATERIAL AND METHODS: For the determined by HPLC-DAD using EBGT solutions bioassay test were used 10 exemplars of nauplius (in prepared at 3.2 mg mL-1 in triplicate. Jejunal rat tissue the microcrustacean’s larval phase) created in NaCl 30 segments were exposed to solubility enhancers g.L-1 solution. The experiment was executed in solution (HP-β-CD 10 and 20%) and monitored for triplicate using unha de gato extracts in 0,025 mg.mL- 120 min, when mounted into the MTS-Snapwell 1 ; 0,05 mg.mL-1 ; 0,1 mg.mL-1 ; 0,2 mg.mL-1; 1 system. TEER measurements were taken for exposed mg.mL-1; 2 mg.mL-1; 10 mg.mL-1; 20 mg.mL-1 and tissues and compared to physiological media (KBR 200 mg.mL-1 concentrations, diluted in saline control). Data were statistically analyzed by t-test, solution. The dead nauplius were counted after 24 ANOVA and Tukey HSD test. Results: The overall Pn hours of samples exposure. As negative control saline and Bn dissolved contents ranged from 3.7 ± 0.13 to solution was used and 1% sodium hydroxide was used 9.9 ± 0.15 µg/mL and 5.5 ± 0.17 to 17.7 ± 0.13 µg mL- as positive control. The test was considered effective 1, respectively. Remarkable aqueous solubility when negative control mortality didn’t surpass 50% improvement of Pn and Bn was significantly seen (p < and when positive control mortality presented values 0,05) for HP-β-CD 20% when compared to other higher than 80%. RESULTS: The negative control solubility enhancers. Furthermore, tissues exposed to presented low mortality level for larvae (47%) and HP-β-CD at 10 and 20% (31 ± 11 Ω.cm2, n = 26) positive control presented high mortality level (87%). showed no statistically difference to control group (29 In 2 mg.mL-1; 10 mg.mL-1; 20 mg.mL-1 and 200 ± 7 Ω.cm2, n = 16) showing no influence on jejunal mg.mL-1 concentrations, the microcrustaceans tissue viability (p > 0,05). Conclusion: The results mortality were 100%. In 0,025 mg.mL-1; 0,05 mg.mL- have provided important data of Pn and Bn solubility 1; 0,1 mg.mL-1; 0,2 mg.mL-1 and 1 mg.mL-1 and jejunal tissue viability in physiologically relevant concentrations, the microcrustaceans mortality were media for application in permeability studies. presented in lower percentage. CONCLUSIONS: The Financial support / Acknowledgments: CNPq, Artemia salina bioassay test presented sensibility in CAPES, FAPEG, FINEP. differents U. tomentosa extracts, proving its toxicity in these concentrations. Financial support/Acknowledgments: Higher Education Personnel Improvement Coordination (CAPES); Master's program in Pharmaceutical ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 202

Sciences of University of Western Paraná; Laboratory 100% for the microcrustaceans. In the other of Cellular Toxicology, State University of Western concentrations, mortality was also verified, though in Paraná. lower percentage. CONCLUSIONS:The bioassay of toxicity on Artemia salina presented sensibility of the microcrustacean to the different samples of sucupira, OT 19 - EVALUATION OF THE TOXICITY OF proving its toxicity in these concentrations. SUCUPIRA (PTERODON EMARGINATUS Financial support/Acknowledgments: Higher VOGEL) ON ARTEMIA SALINA Education Personnel Improvement Coordination (CAPES); Master's program in Pharmaceutical KOERICH, S.1,2; INDRAS, D. M.1,2; REOLON, Sciences of University of Western Paraná; Laboratory A.1,2; CAZ, P.1,2; MAREK, C. B.2; ITINOSE, A. of Cellular Toxicology, State University of Western M.3 Paraná.

1Master's program in Pharmaceutical Sciences of University of Western Paraná, 85819110, Cascavel- OT 20 - IN SILICO TOXICOPHORICAL PR, Brazil; 2Laboratory of Cellular Toxicology, State ANALYSIS OF MAJOR CHEMICAL University of Western Paraná, 85819110, Cascavel- COMPOUNDS FROM ESSENTIAL OIL OF PR, Brazil; 3Assistance Center in Toxicology HYDROCOTYLE UMBELLATA L. (CEATOX), Hospital University of Western Paraná, (ARALIACEAE) UNDERGROUND PARTS. 85806470, Cascavel-PR, Brazil. OLIVEIRA, T. L. S. 1,4; OLIVEIRA, M. G. 2; INTRODUCTION: Medicinal plants are largely used SILVA, V. B. 3; MORAIS, S. R. 1,4; COSTA, E. A. 2; to therapeutic purposes due to their low cost and easy PAULA, J. R. 5 access. The species Pterodon emarginatus Vogel, popularly known as sucupira, is an arboreal species 1 Faculdade de Farmácia da Universidade Federal de whose bark produces a volatile and aromatic oil, Goiás, Goiânia – GO, Brasil; 2. Instituto de Biologia efficient for rheumatism treatment. As the bioactive da Universidade Federal de Goiás, Goiânia – GO, compounds found in plant extracts may present toxic Brasil; 3. Escola de Ciências Médicas, Farmacêuticas e effects, the toxicity tests enable the evaluation of their Biomédicas da Pontifícia Universidade Católica de effects in biological systems. A manner to evaluate the Goiás, Goiânia – GO, Brasil; 4. Universidade Paulista toxicity in plant extracts is through bioassays. Among – UNIP, Goiânia – GO, Brasil. these, the toxicity test about Artemia salina is mentioned, a microcrustacean that lives in saltwater. Introduction: In silico toxicophorical analysis is one The simplicity, the quickness and the low cost favor its type of toxicity assessment that uses computational use in preliminary analyses of general resources instead of animal models as well as to guide toxicity.OBJECTIVE:The present study verified the experimental in vivo assays. Hydrocotyle umbellata L. mortality percentage of Artemia salina on different have been used in folk medicine, however concentrations of plant extracts from sucupira. toxicological studies in support of this medicinal use of MATERIAL AND METHODS: For the bioassay, 10 this species are not clarified. Objective: To investigate exemplars in the microcrustacean’s larval phase of the in silico toxicological potential of major nauplius, created in NaCl 30 g.L-1 solution, were used. compounds found in H. umbellate essential oil of in The experiment was executed in triplicate using plant order to screen potential hazards. Materials and samples of sucupira in the concentrations of 200 Methods: Essential oil was obtained from underground mg.mL-1; 20 mg.mL-1; 10 mg.mL-1; 2 mg.mL-1; 1 parts of H. umbellata submitted to hydrodistillation, mg.mL-1; 0,2 mg.mL-1; 0,1 mg.mL-1; 0,05 mg.mL-1 chemically characterized and quantified by gas and 0,025 mg.mL-1, diluted in saline solution. The chromatography coupled with mass spectrometry dead nauplii were counted after 24 hours of exposure (GC/MS). Retention indices were calculated and with to the samples. Negative control with saline solution the mass spectra, were compared with literature values and positive control with 1% sodium hydroxide were for identification of essential oil components. Toxicity used. The test was considered effective when the prediction of ethylvanillin, β-Selinene and β- mortality of negative control didn’t surpass 50% and Chamigrene 2D structures was performed with ACD/I- the positive one presented values higher than 80%. Lab, Lazar, Pred-hERG and Pred-Skin online tools, RESULTS:The tests showed that in the tubes which are based on chemical similarity criteria with containing saline (negative control), there was low known toxic compounds. Results: The major mortality level for larvae (30%), and the tubes constituents of H. umbellata essential oil were containing positive control presented high mortality ethylvanillin (21.59%), β-Selinene (13.36%) e β- level (97%). It was verified that in the 2 to 200 Chamigrene (12.19%). Prediction data revealed no mg.mL-1 concentrations there was a mortality level of toxicological potential regarding Ames toxicity, hERG ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 203 inhibition and binding to the estrogen receptor. In contrast, a predicted risk of carcinogenicity was detected for the three compounds and a risk of skin sensitisation was predicted for β-Selinene and β- Chamigrene. The analysis of adverse effects on a number of organs and their systems within the therapeutic dose range suggest that β-Selinene and β- Chamigrene present the higher probability to cause adverse effects on blood, cardiovascular system, gastrointestinal system, kidney, liver and lungs than ethylvanillin. Furthermore, the predicted oral lethal dose 50% (LD50) values revealed that β-Selinene and β-Chamigrene present lower LD50 when compared with ethylvanillin. Conclusion: This study contribute to knowledge of toxicological potential of the compounds identified in the essential oil of H. umbellata and could be studied in vivo by animal model. Financial support/Acknowledgments: CNPQ, CAPES, FAPEG

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SC 01 – PROGRAM FOR PREVENTION OF instrument in drug prevention, in reducing school DRUG ABUSE AND VIOLENCE "ANJOS DA violence due to drug abuse. GUARDA" 2016 Financial support / Acknowledgments: FUNDIBIO IBB UNESP FARIA, M.L.V.C1; SANTOS, B.B1; GOMES, A.C.C1; LIMA, C.S1; ANSELMO, F²; OLIVEIRA, G.P1; SENSULINI.L, A1; SILVA, D.P1; GODINHO, SC 02 - LEVELS OF REDUCED GLUTATHIONE A.F1; SANDRIM, V.C2; SILVA, E.J.R2; ALMEIDA, IN CRACK USERS DURING HOSPITAL A.A1. DETOXIFICATION

¹Centro de Assistência Toxicológica (CEATOX), TISSIANI, A, C. 1; FELIPPIN, T. 1; GELATTI, G. T. Instituto de Biociências de Botucatu – UNESP, 1; MAYER, M. S. 1; GOULART, J. S. 1; Botucatu SP. Brasil; ²Departamento de Farmacologia, HAMERSKI, H. M. 1; SALAZAR, R. S. F. 1; HORN, Instituto de Biociências de Botucatu – UNESP, R. C. 1. Botucatu SP. Brasil. 1. Universidade de Cruz Alta. Introduction: Numerous events related to frequent drug abuse and early violence among young people are Introduction: Psychoactive drugs such as crack, are reported. Considering the risk factors for drug use and known by causing the increase of reactive species and abuse, we highlight the lack of information on the change the behavioral functions of the organism. Thus, hazardous effects of drugs of abuse, such as marijuana antioxidants such as Reduced Glutathione (GSH) have and tobacco in the body. The Program for Prevention the ability to eliminate reactive species (ERs) that of Drug Abuse and Violence (PPDAV) “Anjos da cause oxidative stress when in excess. Objective: To Guarda”, developed by CEATOX-IBB/UNESP, verify the levels of GSH in users of crack in a hospital tackles this issue and facilitate spreading information detoxification. Material and Methods: The population about the risks of drug abuse to the population. of this study was composed of 40 crack users in Objective: here we show the results of the PPDAV in treatment for hospital detoxification. Two blood the year of 2016. Methods: we trained undergraduate samples were collected (at the beginning and at the end students in Biology, Biomedicine, Nutrition Sciences of the hospitalization period), with an average interval and Medical-Physics to participate as lecturers in of 20 days among them. The control group consisted of seminars and ludic activities about drugs of abuse to healthy non-drug users. The results were submitted to teachers, elementary, middle and high school students the one-way Analysis of Variance followed by Tukey and general population, with lectures on the harmful test, considering significantly different means with effects of drugs. In these anti-drug sensitization p≤0.05. Results: We observed an increase of 36.52% lectures we demonstrated the harmful effects of drugs GSH levels in crack users at the beginning of of abuse through anatomical-pathological pieces, hospitalization in relation to the control group. At the videos, and drug detection kits in the urine. The end of hospitalization, there was a reduction in GSH lectures for elementary and middle school students levels of 56.33% in relation to the initial period of were held for 60 min/week for each type of drug. hospitalization. Conclusion: The increase in the levels Results: students from the university units of the of GSH observed in the first collection, is probably due Botucatu Campus of UNESP were included. The main to the endogenous response to the aggression caused target audience for our activites were middle and high by crack consumption, thus it is increasing the school students (n = 740) from public (06) and private antioxidant defenses. On the other hand, as a result of (02) schools; Institutions such as SENAC (n=80), hospital detoxification, GSH levels decreased due to Churches (n=100), Narcotics and Alcoholics the use of this antioxidant in order to reduce the Anonymous (n = 15), Students of the EMBRAER generation of ERs produced by the use of Crack and/or College (n=600), young (n=18) of the Central of to reverse the oxidative damages caused by the action Measures and Alternative Feathers (CPMA)1º Vara of these species on lipids, proteins, and ADN. Fórum Botucatu SP, University students (n=) from , Financial support / Acknowledgments: UNICRUZ and attendees of the “Meet the IBB” event (n=800). Conclusion: In 2016, almost 1000 students and school teachers attended the events promoted by our drug- prevention program. These attendees could take the information they learn to their families, thus spreading important information that may help the reduction of drug-related violence events in their communities. We highlight the importance of the project as an essential

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SC 03 - ADHESION TO PHARMACOLOGICAL SC 04 - ASSESSMENT OF THE USE OF DRUGS TREATMENT OF MENTAL DISORDERS FROM AND OTHER SUBSTANCES TO IMPROVE ALCOHOL ABUSE IN PATIENTS OF COGNITIVE PERFORMANCE PSYCHOSOCIAL ATTENTION CENTER (CAPS) OF CRUZ DAS ALMAS – BA BAZARIN, J. 1; TOLEDO, O. R. 1; SUCHARA, E. A. 1. CRUZ, R. C. S. 1; TELES, A. L. B. 2. 1 Universidade Federal de Mato Grosso, Campus do 1. Faculdade Maria Milza, Governador Mangabeira - Araguaia, Barra do Garças - MT, Brasil. BA, Brasil; 2. Universidade do Estado da Bahia, Salvador - BA, Brasil. Introduction: Many substances have been used to improve the cognitive capacity, consequently INTRODUCTION: The development of mental and improving academic performance. Objective: The goal behavioral disorders arrising from alcohol abuse its a of this study was to assess the use of drugs and other very common health problem observed in 20% of the substances to improve cognitive performance in a Cruz das Almas city (Bahia, Brazil) Psychosocial sample of college students. Materials and Methods: Attention Center (CAPS) patients. In most cases, the This is an observational cross-sectional pharmacological treatment of these disorders are based epidemiological study conducted in public and private on antidepressants (AD), antipsychotics (AP) and colleges of Barra do Garças, State of Mato Grosso, mood stabilizers (MS). OBJECTIVE: The purpose of Brazil, from May to June 2015. Statistical analyzes this study (Approved by the Faculdade Maria Milza’s were performed using Epi Info version 3.5.2 ® Research Ethics Committee, protocol number: program. Results: A total of 1,165 questionnaires were 043/2012) was to evaluate the adherence to applied in public and private colleges. Most pharmacological treatment with AD, AP and MS drugs participants were female aged between 20 and 24 on mental disorders resulting from alcohol abuse, in years. It was observed that most of the students had patients from Cruz das Almas CAPS. MATERIAL already used substances to help them study, some of AND METHODS: A semi-structured interview based those being drugs, such as methylphenidate on the Morisky Medication Adherence Scale was hydrochloride, fluoxetine, and piracetam, as well as applied during September and October 2012 to patients other substances, such as caffeine, energy drinks and who used at least one of the refered drug classes for guarana powder (Paullinia cupana). The use of drugs the treatment of mental disorders due to alcohol abuse. was reported by students from the Biological Sciences The subjects of the study were classified as having and Health, Human and Social Sciences, and Exact high (HAd) or low degrees of adherence (LAd). Sciences and Earth fields. Most of the students that RESULTS: 65% of the 20 patients interviewed showed used other substances were from the Exact Sciences HAd. In the Morisky questions, forgetfulness was the and Earth field, followed by those from the Biological most frequent (70%) among LAd patients. In addition, Sciences and Health, and Human and Social Sciences some socio demographic factors appears to be fields. Only 21.4% of the students noticed differentiated between the two groups. HAd patients improvement in academic performance using other had 46% of subjects in the 50 to 59 years age range, substances, while 75.7% noticed improvement with the compared to 28% in the LAd group, among the latter use of drugs. Conclusion: It was concluded that college presenting 43% of patients in the 30 to 39 years age students used drugs and other substances to improve range. For the HAd patients, in turn, the 30 to 39 years their academic performance. This is a risk factor, age range represents only 15%. 57% of patients with because these drugs and substances have adverse LAd have already used illicit drugs, compared to 7% impacts or are consumed with no medical among patients with HAd, demonstrating a clear prescriptions. Therefore, there is an urgent need of contrast. There is a higher frequency of unemployed promoting effective actions to alert and raise patients (70%) and that complain of side effects (71%) awareness among college students about the among LAd patients with relation to the HAd group consequences of using these drugs and substances. (7% and 53% respectively). CONCLUSION: Some Financial support / Acknowledgments: Universidade possible factors of influence in the adherence to the Federal de Mato Grosso - Campus do Araguaia pharmacological treatment by the subjects of the research, as age and side effects, may contribute to guide actions performed by the CAPS of Cruz das Almas to increase the adherence of patients to the pharmacological treatment of their mental disorders. Financial support / Acknowledgments: UNEB

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SC 05 - EVALUATION OF THE SC – 06 AYAHUASCA: OBSERVATIONAL EPIDEMIOLOGICAL PROFILE OF DRUG STUDY OF THE EFFECTS PRESENTED IN INTOXICATION IN BRAZIL, FROM 1999 TO USERS IN THE MUNICIPALITY OF 2013 ABADIÂNIA-GO

CHEQUER, F.M.D. 1,2; ANASTÁCIO, L.B. 1; FERREIRA, F.S.; SOUZA, K.A.; VASCONCELOS, SOARES, L.S. 1; BALDONI, N.R. 3. F.G.

1. Universidade de Itaúna, Itaúna - MG, Brasil; 2. Centro Universitário de Anápolis - UniEvangélica, Universidade Federal de Minas Gerais, Belo Horizonte Anápolis - GO, Brasil. - MG, Brasil; 3. Universidade de São Paulo, Ribeirão Preto - SP, Brasil. Ayahuasca is a decoction made of two plants. A lily of the family Malpighiace, called Banisteriopsis caapi Introduction: The use of drugs of abuse is responsible and leaves of the family Rubiaceae being the plant for several hospitalizations in the national territory. Psychotria viridis. It’s popularly used in the Saint However, the epidemiological profile of these Daime’s religion, considered a psychoactive drink, intoxications is still nebulous, fomenting researches to inhibitor of the enzyme Monoamino Oxidase - MAO, outline the profile of such intoxications. Objectives: To thus enabling the action of tryptamine evaluate the epidemiological profile of drug Dimetiltryptamine - DMT, as a serotonergic agonist. intoxication in Brazil. Method: Research in the Due to its mechanism of action, it’s a potent National System of Toxic-Pharmacological hallucinogen, has physical effects such as vomiting, Information (SINITOX) on intoxication by drugs of nausea and diarrhea, and it also highlights as main abuse. The following parameters were collected: effects increased concentration, elevated self-esteem, circumstance, age, sex, zone, evolution of the great access to autobiographical memories, increased condition and death, during the period from 1999 to sensitivity and empathy. These effects being the reason 2013. Results: It was observed that intoxication by why it’s sought and used. The objective of this work is drugs of abuse is generally associated with accidental to conduct an observational study of effects caused by intoxication due to abuse (84.28%) and attempted self- use of ayahuasca tea, used in the Saint Daime’s extermination. In relation to the profile of the religion, taking into account effects already portrayed intoxicated, it is noticed that the intoxicated individual by literatures. This is a qualitative observational study, is young, predominantly with 20-29 years old carried out in the community of Saint Daime in (32,97%). In addition, intoxications occurred Abadiânia-GO. It was did an observation of the effects predominantly in males (74.40%) and in urban settings visibly presented after the use of ayahuasca by 45 (83.79%). Observing the evolution of the cases of users during a ceremony. As results it has been intoxication, it was noticed that most of the observed that users manifested several reactions. The intoxications evolved to cure (68.96%), and the most frequent were nausea followed by vomiting, number of patients that have sequels or have died after ecstasy and trance state, in addition to these intoxication (2.34%) less expressive. In terms of the manifestations, some users show a feeling of intense analysis of the progression of intoxicated patients, cold. In a more isolated way, some users also there was an expressive increase (from 1999 to 2013) presented other types of reactions like easy laughing in the incidence rates of intoxication by drugs of abuse without apparent reason, stereotyped movements, in the age groups from 14 to over 80 years, mainly pallor and red eye conjunctiva. Although ayahuasca is after 2009. In mortality rate, there was a low variation used with a religious doctrine purpose, the results between the years studied, with an increase in this rate showed that its use is capable of cause different in the year 2003 and its decrease after the year 2012. reactions in the user's body, reactions that are at risk of However, it is necessary to evaluate that most of the possible harm to these people. Taking into account statistics offered by SINITOX are derived from the what has been observed, these reactions are clearly Information and Assistance Centers (CIAT), which compatible with those attributed in other articles, thus have decreased their participation in these surveys. demonstrating their potential for action. Thus, it is Thus, the numbers of deaths can be more expressive, necessary to carry out other studies on toxicity, in fact. Hence, it was concluded that the profile of especially among users who use continuous, in order to intoxicated by drugs of abuse in Brazil is currently the have proof of their safety of the use of Ayahuasca. young man, who lives in urban areas, is accidentally intoxicated and is able to evolve the intoxication to the cure. Financial support / Acknowledgments: Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) and Universidade de Itaúna (UIT). ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 208

SC 07 - RELATIONSHIP BETWEEN LEVELS OF SC 08 - LEVELS OF MALONDIALDEHYD IN OXIDATIVE STRESS MARKERS IN CRACK CRACK USERS DURING HOSPITAL USERS WITH THE NUMBER OF STONES USED DETOXIFICATION¹ PER EPISODE GOULART, J. S.; HAMERSKI, H.M. 1, GELATTI, GELATTI, G. T. 1; FELIPPIN, T. 1; MAYER, M. S. G.T. 1, FELIPPIN, T. 1, TISSIANI, A.C. 1, MAYER, 1; TISSIANI, A. C. 1; GOULART, J. S. 1; M.S. 1, GARCES, N.B. 1 , SALAZAR, R.F.S.S. 1, HAMERSKI, H. M. 1; SALAZAR, R. F. S. 1; HORN, HORN, R.C. 1. R. C. 1. 1Laboratory of Medicinal Plants and Oxidative Stress 1 Laboratory of Medicinal Plants and Oxidative Stress (LAMOX), Research Group on Comprehensive Health (LAMOX), Research Group on Comprehensive Health Care (GPAIS), University of Cruz Alta (UNICRUZ) - Care (GPAIS), University of Cruz Alta (UNICRUZ). RS, Brasil.

Introduction: The abusive crack consumption is Introduction: Crack is the third most commonly illegal associated to physical, psychiatric and social problems. substance in Brazil and its high commercialization In addition, studies suggest that the administration of classifies it as a pandemic situation and it is considered crack increases the brain dopamine levels, it results in a public health problem. The withdrawal symptoms a high production of reactive oxygen species, which during hospital detox cause increased levels of may lead the user to the oxidative stress. Objective: To oxidative markers such as malondialdehyde (MDA), a correlate levels of oxidative stress markers in crack lipid peroxidation product, that may be provoked by users according to the number of “crack stones” used the exposure to psychoactive drugs. Objective: To per episode. Material and Methods: The sample was evaluate MDA levels in crack users during hospital consisted of 40 male crack users in a hospital detox. detox. Material and methods: The audience of this These users were divided into three groups according study was composed of 40 crack users in a hospital to the number of crack stones used per episode: 4-10 detox treatment. Two blood samples were collected, stones, 11-20 stones and more than 20 stones. Blood the first at the beginning and the second at the end of collection and questionnaire application were hospitalization period, with a mean interval between performed on the day of admission. Levels of them of 20 days. The control group consisted of Thiobarbituric Acid Reactive Substances (TBARS), healthy individuals and non-drug users. The results Carbonylated Proteins (CPs) and Reduced Glutathione were submitted to one-way Analysis of Variance (GSH) were measured in the plasma of these subjects. followed by Tukey's test, which was considering This study was approved by CEP of UNICRUZ, RS. significantly different means with a p. Statistical analysis was performed by using Pearson Financial support / Acknowledgments: CAPES – correlation test and one-way ANOVA followed by the CNPq. Tukey test. Results: When it was compared the levels of oxidative stress markers and the number of stones used per episode, we observed high levels of TBARS SC 09 - TOXIC EFFECTS OF HOUSEHOLD in the group of users who used more than 20 stones per CLEANERS episode (r=0.75) compared to other groups and was not correlation in CP and GSH levels. Conclusion: The PIERRO, J. R. 1; MACHADO, M.1; SOUSA, N.1; higher the number of crack stones consumed by PINHEIRO, F.1. episode, higher are the TBARS levels. The predominance of the action of crack on lipids can be 1 INTERTOX Ltda. São Paulo – SP, Brasil. justified by its high affinity for liposoluble compounds, which increases the tendency of this xenobiotic to Introduction: Household cleaners is a term used to bioaccumulate in tissues rich in fatty acids and, identify sanitizers intended for home use, being consequently, to induce the occurrence of oxidative substances or preparations for sanitizing, disinfecting stress in these tissues. or disinfestation at home. Studies have shown that Financial support / Acknowledgments: UNICRUZ these products are responsible for many cases of poisoning. In Brazil, the registration of household cleaning products is regulated by ANVISA (National Health Surveillance Agency), which assesses and manages product risk prior to consumer release. Objective: To present and discuss toxic effects to the health of household cleaning products, aiming to obtain a balance between the beneficial effects promoted by the use, and the measures of prevention ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 209 and control of the possible harmful effects of the sanitary products, that in their compositions may chemical substances. Materials and Methods: contain caustic and harmful substances to the health of Theoretical studies on the toxic effects of substances in the user. Objective: To discuss chemical intoxications household cleaning products were carried out based on by household products in Goiás (2011- 2015). scientific references from internationally recognized Methodology: This is a descriptive epidemiological databases. Based on this information, standards such as study, whose data were obtained by consulting the ABNT / NBR 14725 and the GHS (Globally SINAN (Notification of Injury Information System) Harmonized System) by UN (United Nations) were database available By the Department of Informatics consulted. Results: The analysis of the product showed of the Unified Health System (DATASUS), at the that some household cleaning substances such as electronic address (http://www.datasus.gov.br). We hydrogen peroxide, sodium hypochlorite, sodium used the free Geographic Information System (GIS) hydroxide, lilial and citral have the potential of causing QGIS and performed geoprocessing techniques to damage to health. These substances have been subject visualize, edit, create and analyze the reference map to the hazard classification criteria for mixtures data. The baseline and thematic maps were elaborated established by the GHS and have resulted in the with the symbologies of cartographic normalization of following hazard classifications of the finished the Technical Manual of Cartographic Conventions of products: corrosion and skin and eyes irritations, skin the Brazilian Army as a guide. RESULTS: It was sensitization, reproductive toxicity, respiratory tract observed a prevalence of intoxications by household irritation and damage neurological functions. products of 44.55% of total exogenous intoxications. Conclusion: Although the release of the sale of these Household products were the most prevalent in 2011, products and the non-mandatory reporting of these 2012, 2014 and 2015 with 43.63%, 52.41%, 47.10% toxic effects, this study showed that the products may and 49.66%, respectively. The counties that presented have potential to cause health damage depending on the most notifications were Goiânia, Formosa, the time of exposure and the inappropriate use that Anápolis, Rio Verde, Valparaiso de Goiás and Jataí. often occurs. Thus, it was concluded that despite the There was a higher prevalence in the female gender beneficial effects promoted by the use of these (53.23%). The highest indexes are in the age groups of products for hygiene and disinfection, these products 1-4 years (59.17%) and < 1year (58.21%), being deserve greater attention because they may present mostly accidental. Conclusion: The results obtained possible toxic effects. conclude that the population, for the most part, are Financial support / Acknowledgments: INTERTOX unaware of the toxic properties of the components in Ltda. São Paulo – SP, Brasil. the products they are using. In addition, the trivialization, as a consequence of the lack of information, of the consumption of these chemicals in SC 10 - EPIDEMIOLOGICAL PROFILE OF the domestic environment, as well as the continuous INTOXICATIONS CAUSED BY HOUSEHOLD and simultaneous use of several of them, mainly by PRODUCTS REPORTED IN THE STATE OF women, shows a high health risk and could lead to GOIÁS IN THE YEARS 211 TO 2015 intoxication. Financial support / Acknowledgments: Conselho SILVA, L. A. 1; PRADOS, V. O. 2; MACHADO, H. Nacional de Desenvolvimento Científico e C. P. 3; NASCIMENTO, H. A. 4; SILVA, L. O. 5; Tecnológico (CNPq). REZENDE, G. M. S. 6; OLIVEIRA, R. R. 7; MAYA, A. C. L. 8. SC 11 - EPIDEMIOLOGICAL PROFILE OF 1 Centro Universitário de Anápolis, Anápolis - GO, ABUSE DRUG INTOXICATIONS REPORTED IN Brasil; 2. Centro Universitário de Anápolis, Anápolis - THE STATE OF GOIÁS BETWEEN 2011 AND GO, Brasil; 3. Centro Universitário de Anápolis, 2015 Anápolis - GO, Brasil; 4. Centro Universitário de Anápolis, Anápolis - GO, Brasil; 5. Centro SILVA, L. O. 1; MARTINS, I. L. O. 1; LIMA, G. C. Universitário de Anápolis, Anápolis - GO, Brasil; 6. 1; VASCONCELOS, F. G. 1; OLIVEIRA, R. R. 1; Centro Universitário de Anápolis, Anápolis - GO, MORAIS, L. T. G. 1; SILVA, L. A. 1; Brasil; 7. Centro Universitário de Anápolis, Anápolis - NASCIMENTO, H. A. 1; REZENDE, G. M. S. 1. GO, Brasil; 8. Centro Universitário de Anápolis, Anápolis - GO, Brasil. 1 Centro Universitário de Anápolis

Introduction: Exogenous intoxications are a set of Introduction: The study of the epidemiological profile adverse effects produced by a chemical (or physical) of intoxication in a given region aims at obtaining na agent, due to its interaction with the biological system. full understanding of the potentially inductive scenario Among these intoxications are the ones by domestic of this state. Thus, the study of the factors that lead to ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 210 intoxication by drugs of abuse is necessary since it is a Introduction: Although the main victims of accidents pertinent issue of public health in Goiás. Objective: To with poisonous animals are men living in rural areas, investigate the data on intoxication by drugs of abuse about 16% of them are children. In these cases, in the State of Goiás between 2011 and 2015. children represent greater severity than adults and Methods: Descriptive, quantitative, retrospective study should receive special monitoring, considering the of data from the website of the Department of concentration of poison by body area, the child's low Informatics of SUS. Among the variables studied are immunological system and lower muscle mass. age, county of registry and circumstance. Results: The Objective: To characterize child accidents by county of Goiânia has 29.5% of the records. Then poisonous animals registered in an information and there are Valparaíso de Goiás (25.7%) and Jandaia toxicological assistance center of Northwest Paraná. (6.04%). The most affected age group was 20 to 39 Material and Methods: Descriptive and documentary years (49.18%), followed by 40 to 59 years (27.19%) study, based on consultation in the epidemiological and 10-19 years (15, 42%). The most prevalent records of notification and care of children from zero circumstances were drug abuse (57.47%), more to 12 years, from 2013 to 2015, registered at the prevalent in the 20-39 age group (48.57%), followed Poison Control Center of the University Regional by regular use (22.56%) also in the previously Hospital of Maringá. The data were transcribed to mentioned age group. The number of cases of drug worksheet in Microsoft Excel® 2013 and the results abuse in youth in Goiás reflects the public health were analyzed descriptively. Results:304 problem in Brazil in which many individuals, epidemiological notification records and assistance of especially Young people, become drug-dependent and children victims of accidents by poisonous animals suffer the consequent risks of these substances. Drug were found, with a predominance of accidents in males use usually begins in adolescence, with licit products (53.9%), age under five years (63.8%) and residents of such as alcohol. The progression to illicit drugs is due the urban area (78.3%). Accidents were mainly caused to peer pressure, especially in younger people. by unidentified insects (32.7%), spiders (22.8%) and Research indicates a significant relationship between scorpions (19.1%). Hospitalizations lasted, on average, drug abuse and violence, accidents and traumas, and two days and most cases were considered light consequently mortality. This behavior tends to increase accidents. The severity of scorpions, with a long period the possibilities of intoxication, also caused by of hospitalization, and the infectious complications stressful routine, peculiarities of the group of friends caused by "unidentified" insects, and seasonality were and the attendance of environments that induce the highlighted - more hospitalizations in the summer and onset of alcohol consumption. Conclusion: Based on spring time. Conclusion: The profile of accidents with this information, it is possible to infer that the State of poisonous animals in children showed a higher number Goiás faces a challenging reality for public agencies, of cases in children younger than five years of age and especially the health system, which must be trained in living in the urban area of the Northwestern Health order to meet the demand presented by the registries Macroregion of Paraná. The present study can since the service to the intoxicated Drugs is complex contribute to the improvement and development of and requires efficacy and expertise. systematization for assistance of children victims of Financial support / Acknowledgments: Centro poisonous animals, as well as conduct educational Universitário de Anápolis actions that reduce the incidence of these accidents and guide first aid to these victims.

SC 12 - ACCIDENTS BY POISONOUS ANIMALS IN CHILDREN REGISTERED IN AN SC 13 - HEALTH BELIEF MODEL AND INFORMATION AND TOXICOLOGICAL POSSIBILITIES OF CHANGES IN THE ASSISTANCECENTER ROUTINE OF FAMILIES OF INTOXICATED CHILDREN GUEDES MRJ1; SALES CCF2; TEIXEIRA RHA3; GONÇALVES KM4; KITAGAWA T5; OLIVEIRA GUEDES MRJ1; MARCON SS2; OLIVEIRA MLFO6. MLFO3.

1 Hospital Universitário Regional de Maringá, 1 Hospital Universitário Regional de Maringá, Maringá, Paraná, Brasil; 2 Hospital Universitário Maringá-Paraná, Brasil; 2 Universidade Estadual de Regional de Maringá, Maringá, Paraná, Brasil; 3 Maringá, Maringá- Paraná, Brasil; 3 Universidade Universidade Estadual de Maringá, Maringá, Paraná, Estadual de Maringá, Maringá- Paraná, Brasil. Brasil; 4 Universidade Estadual de Maringá, Maringá, Paraná, Brasil; 5 Hospital Universitário Regional de Introduction: Child intoxications are often accidental Maringá, Maringá, Paraná, Brasil; 6 Universidade ingestion of products found in the home, and occur Estadual de Maringá, Maringá, Paraná, Brasil. because children have characteristics that make them ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 211 more vulnerable to accidents. Objective: to identify withdrawal throughout the detoxification process may changes in the routine of intoxicated children attended lead the users to increased levels of markers of at an emergency unit, with a view to acquiring oxidative stress in their bodies. Proteins are among the preventive behaviors. Material and Method: an main targets of oxidants due to their high speed for observational study, in the light of the Rosenstock various reactions with reactive species and their Health Belief Model, carried out with family members abundance in biological systems. Objective: To of 11 intoxicated children attended in the stabilization evaluate the levels of protein carbonyls (PCs) in crack room and in the pediatric intensive care unit of a users in a hospital detox. Material and Methods: The teaching hospital in the Northwest of Paraná. Data target audience of this study was composed of 40 users collection took place from March to October 2015, of crack in treatment for hospital detox. Two blood through documental analysis, and observation with samples were collected, the first at the beginning of moderate participation and home interview. The data hospitalization and the second at the end, with a mean sources were the Toxicological Occurrence form of the break between them of 20 days. The control group was Poison Control Center. The interviews were recorded consisted of healthy individuals and non-drug users. and the statements were transcribed and analyzed The results were submitted to the one-way Analysis of through the analysis of thematic content, according to Variance followed by Tukey's test, which was the dimensions: perceived susceptibility and severity, considered significantly different means with a p. and benefits and barriers to prevention. Results: 11 Financial support / Acknowledgments: Universidade children were hospitalized, nine were male, up to two de Cruz Alta years old, with individual accident, occurred in the house and in the presence of the parents. Six cases were clinically classified as were users of the basic SC 15 - MORBIMORTALITY FOR SUICIDE health units as a single reference for health care. AND SUICIDE ATTEMPTS AND USE OF Conclusion: the analysis of the dimensions of the CHEMICAL AGENTS, PARANÁ-BRAZIL Health Belief Model indicated that the susceptibility to household intoxication was perceived, but the severity SUGUYAMA, P. 1; GUEDES, M. R. J. 2; SILVA, J. of the event was neglected by the family. Also, the S. 3; HEBERLE, J. T. 4; OLIVEIRA, M. L. F. 5. benefits of preventive behavior were not perceived and families reported barriers related to health services, 1 Universidade Estadual de Maringá - PR, Brasil; 2 especially access and bonding with health Hospital Universitário Regional de Maringá - PR, professionals. Household actions of primary care Brasil; 3 Hospital Universitário Regional de Maringá - professionals were not reported by any family, who PR, Brasil; 4 Hospital Universitário Regional de indicated social pressure from the school, and Maringá - PR, Brasil; 5 Universidade Estadual de criticisms of the health service and the family itself Maringá - PR, Brasil when intoxication occurred. The results point to the need for actions and home support, as it was verified Introduction: health information systems are important that intoxication did not act as a "turning point" for tools that help understanding the health problems of changes in the household and social environment of the population. Objective: to analyze the profile of the families after hospital discharge. suicide and suicide attempt – TS by chemical agents in the state of Paraná - Brazil. Methods: ecological study, with data from the Grievance Notification Information SC 14 - PROTEIN CARBONILLATION LEVELS System (SINAN) and from the Mortality Information IN CRACK USERS DURING HOSPITAL System (SIM), accessed in July 2017. Confirmed DETOXIFICATION intoxication cases from SINAN were considered; and in the SIM, the cases that presented as the basic cause MAYER. M. S.; MAYER, M. M. 1; FELIPPIN, T J. 1; of death ICD 10: X60 to X69. All notifications GELATTI, G. T. 1; TISSIANI, A. C. 1; GOULART, J. available for access in the period from 2010 to 2015 S. 1; HAMERSKI, H. M. 1; SALAZAR, R. F. S. 1; were included. Microsoft Excel® was used for data HORN, R. C. 1. tabulation and descriptive analysis. Results: 21,790 cases were reported in the SINAN and 521 in the SIM, 1 Laboratório de Plantas Medicinais e Estresse with an annual average of 3,632 TS and 87 suicides, Oxidativo (LAMOX), Grupo de Pesquisa em Atenção with an increasing trend in the analyzed period. In Integral a Saúde (GPAIS), Universidade de Cruz Alta SINAN there was predominance of women - 71.3%, (UNICRUZ). with 1.8% pregnant women. In SIM, the male gender was - 61.8%, and 49.6% were married, widowed or Introduction: The consumption of psychoactive divorced. In TS, the prevalence was between 20-59 substances such as crack is considered a public health years of age (71.9%) and in the suicide of 30-59 years problem. The abstinence symptoms generated by drug (64.7%). In relation to the toxic agent, TS was found to ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 212 use medication - 71.4%, and in the suicide the use of notifications by region, with predominance in the the pesticide - 44.7%. There was a heterogeneous Southeast region - 51.7% and fewer cases in the North distribution of notifications according to the region - 2%. Acute intoxications in 82.6% of cases and notification region of the TS, the 2nd Metropolitan acute-repeated 13%. There was cure without sequel in Health Regional presented 33.8% of the cases, 82.6%, but sequels and deaths represented 5,136 cases followed by the 15thHealth Regional Maringá - 9.8%; - 3.4%. Conclusions: there was an increase in reports in suicide at 2nd Metropolitan Health Regional of attempted suicide by chemical agents with the (26.7%) followed by 17thHealth Regional Londrina. advancement of the years, predominantly female, and Conclusions: female morbidity and mortality in males, acute type drug poisonings. The Southeast region occurrences in young adults and the use of medication registered the highest number of notifications. in TS and pesticides in suicide delineate the profile. The importance of health information systems in the profile dimensioning of suicide and TS cases and the SC 17 - COMPARISON OF ALCOHOL AND identification of needs for suicide prevention actions is OTHER DRUGS CONSUMPTION BETWEEN reaffirmed. TEENAGERS STUDENTS IN DIFFERENT SOCIO-OCCUPATIONAL AREAS OF A CITY IN SOUTHERN BRAZIL SC 16 - EPIDEMIOLOGICAL PROFILE OF SUICIDE ATTEMPTS BY INTOXICATION IN MOSSINI, S.A.G.1,2; MUNHOZ, G.R.1; PEREIRA, BRAZIL E.Q.3; APOLINÁRIO, A.P.A.1; MIRANDA, B.C.1; REIS, L.C.1; ZANIN, R.A.4; BANDO, E.1; SUGUYAMA, P. 1; ROSADO, P. V. V. 2; CHRISTOPHORO, R.5; NISHIYAMA, P.1; ANDUJAR, B. T. 3; GUEDES, M. R. J. 4; OLIVEIRA, M.L.F.5. OLIVEIRA, M. L. F. 5. 1.Laboratory of Toxicology, Department of Basic 1 Universidade Estadual de Maringá - PR, Brasil; 2 Health Sciences, State University of Maringá, Maringá Universidade Estadual de Maringá - PR, Brasil; 3 – PR, Brazil; 2.Postgraduate Program on Bioscience Universidade Estadual de Maringá - PR, Brasil; 4 and Physiopathology, State University of Maringá, Hospital Universitário Regional de Maringá - PR, Maringá – PR, Brazil; 3. Department of Statistical, Brasil; 5 Universidade Estadual de Maringá - PR, Postgraduate Program in Biostatistics, State University Brasil of Maringá, Maringá – PR, Brazil; 4. Psychosocial Assistance Center - Alcohol and Drugs (Caps AD), Introduction: The suicide attempt is estimated to be Maringá – PR, Brazil; 5. Department of Nursing, about twenty times more frequent than suicide, and Postgraduate Program in Nursing, State University of that for each recorded attempt there are at least four Maringá, Maringá – PR, Brazil unrecorded suicide attempts. Chemical agents are of great demand for health services. Objective: to analyze Introduction: The combination of genetic, the epidemiological profile of the suicide attempt by psychological, family, socioeconomic and cultural chemical agents in Brazil, from 2010 to 2015. factors influences drug use. Aims: To compare alcohol Methods: ecological study with data from the and other drugs consumption among students from Grievance Notification Information System, accessed different socio-occupational areas in a city in southern in June 2017. Cases of intoxication from all Brazilian Brazil. Methods: This was a descriptive epidemiologic federated units were included, with full years of study of transversal design, performed by random notification available for access. It was used Microsoft sampling stratified by school, with probability Excel® for data tabulation and descriptive analysis. proportional to the number of students allowed to Results: 152,135 cases of intoxication suicide were participate. DUSI (Drug Use Screening Inventory) was reported in the analyzed period, with an annual the tool chosen for data collection. Schools were average of 25,356 notifications, with an increasing distributed in 4 areas (01 - Popular, 02 - Higher trend observed in the period. There was female Middle, 03 - Popular Agrícola and 04 - Superior), predominance - 70.3%, of these women 2,479 - 1.6% according to socio-occupational typology of the were pregnant. As for the age group, the reports of Metropolitan Region of Maringá-PR, defined by young adults (20 to 39 years old) were predominant, Rodrigues and Accorsi within CNPq-INCT- with 70%; in terms of schooling, 45.6% were Observatory of the Metropolis (2009-2013). Statistical blank/ignored, primary education was 27.1%; middle analysis was performed through the R and SAS education 21.6% and higher education 3.9%. programs. Results: Thirty-one schools participated in Regarding to the means used, medication was the most the study and 697 forms were gathered, with age used - 66,6% and the use of pesticides - 21,6% of the ranging from 10 to 19 years old. There were found as cases. There was a heterogeneous distribution of the most consumed psychoactive substances in the 4 ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 213 areas analyzed, alcohol> analgesics> tobacco> collection. Odds ratio was used to analyze the marijuana. Alcohol consumption was higher at the demographic profile and to identify protection factors most socio-occupational privileged areas (2 and 4), to drug use. Statistical analysis was performed through associated with higher analgesics, marijuana, tobacco the programs R and SAS. Results: Thirty-one schools and inhalants consumption. The schools number was participated in the study and 697 forms were gathered, similar only in areas 2 (n = 16) and 4 (n = 11), both of with age ranging from 10 to 19 years old, of which which were hierarchically superior to areas 1 and 3. It 54.95% said already have used drugs. The first contact is important to highlight the higher teenager families has happened between 14 and 16 years old, except for power purchasing who were attending in schools alcohol, that has happened before 11 years old. located in areas 2 and 4, and the higher families Alcohol was the preferred drug (42.43%), followed by schooling which has suggesting that financial analgesics (26.00%), tobacco (13.50%), marijuana availability may exert significant influence over drug (10%) and inhalants (7.50%). The statistical variables, use. Conclusions: This study showed higher alcohol elementary/high school, shift and grade were use in the socio-occupational privileged areas, associated with drugs use, p value. associated with high analgesics, marijuana, tobacco Financial support / Acknowledgments: and inhalants consumption. In spite of the socio- Decit/SCTIE/MS; CNPq; Araucaria Foundation and occupational aspects have been considered as SESA-PR for their support. facilitators, the understanding about the dangers that psychoactive substances consumption can offer appears not to be reach by teenagers, who present a SC 19 - RELATIONSHIP BETWEEN greater number of stories use and experimentation, OXIDATIVE STRESS MARKERS AND when compared to less favored areas. FREQUENCY OF WEEKLY USE OF CRACK Financial support / Acknowledgments: Decit/SCTIE/MS; CNPq; Araucaria Foundation and FELIPPIN, T. 1; GELATTI, G. T. 1; MAYER, M. S. SESA-PR for their support. 1; TISSIANI, A. C. 1; GOULART, J. S. 1; HAMERSKI, H. M. 1; SALAZAR, R. F. S. 1; HORN, R. C. 1. SC 18 - EXPOSURE TO ALCOHOL AND OTHER DRUGS BETWEEN TEENAGERS STUDENTS: 1 Universidade de Cruz Alta, Cruz Alta - RS, Brasil. INVESTIGATION OF CONSUMPTION AND PROTECTIVE FACTORS Introduction: The crack consumption leads us to one of the most severe forms of dependence, due to its quick MOSSINI, S.A.G.1,3; PEREIRA, E.Q.2; access to the pulmonary alveoli, which are highly APOLINÁRIO, A.P.A.1; MUNHOZ, G.R.1; vascularized and propiciate the nervous system MIRANDA, B.C.1; REIS, L.C1; MENOTTI, V.S.1,3; performance in a few seconds. In addition, crack users BANDO, E.1; CHRISTOPHORO, R.4; NISHIYAMA, are susceptible to the oxidative stress condition, which P.1; OLIVEIRA, M.L.F.3 is related to the severity of withdrawal symptoms generated by drugs abscense throughout the detox 1.Laboratory of Toxicology, Department of Basic process. Objective: Match the levels of oxidative stress Health Sciences, State University of Maringá, Maringá markers in crack users according to their weekly use of – PR, Brazil; 2. Department of Statistical, Postgraduate this drug. Material and Methods: The sample consisted Program in Biostatistics, State University of Maringá, of 40 crack users and 20 non-crack users for reference Maringá – PR, Brazil; 3. Postgraduate Program on value fins (18.5 ηmol MDA / mL), all male users in Bioscience and Physiopathology, State University of hospital detoxification. These users were subdivided Maringá, Maringá – PR, Brazil; 4.Department of into three groups as the frequency of weekly crack use: Nursing, Postgraduate Program in Nursing, State 1-3 days, 4-5 days, 6-7 days. Blood collection and University of Maringá, Maringá – PR, Brazil. questionnaire application were performed on the day of admission. Levels of Thiobarbituric Acid Reactive Introduction: First contact with drugs happens in Substances (TBARS), Carbonylated Proteins (CPs) adolescence, period of greater psychological and social and Reduced Glutathione (GSH) were measured in the vulnerability. Aims: To evaluate alcohol and other plasma of these audience. This study was approved by drugs consumption among teenagers of public school CEP of UNICRUZ, RS. Statistical analysis was system of a city in southern Brazil and to identify performed by using Pearson correlation test and one- protective factors. Methods: Descriptive, way ANOVA followed by the Tukey test. Results: We epidemiologic, transversal study, performed by have found a reduction in TBARS levels in individuals random sampling stratified by school, with probability who used crack 4-5 and 6-7 times a week when proportional to the number of students allowed to compared to users who used crack 1-3 days a week participate. DUSI was the tool of choise for data (r=90). Regarding to the levels of PCs and GSH, there ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 214 was no change in the frequency of crack consumption. Conclusion: It is suggested that the behavior observed in individuals who used crack 1-3 days a week had higher levels of TBARS. It might be justified by the neuroadaptation effect of the organism that prevails during the withdrawal syndrome on the days when the drug was not consumed, moreover the physico- chemical properties of crack have shown a high affinity for lipids. Financial support / Acknowledgments: Universidade de Cruz Alta - UNICRUZ

TOXINOLOGY ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 216

TO 01 - GENOTOXICITY, CYTOTOXICITY BjussuLAAO-II induce cell death and DNA damage in AND OXIDATIVE STRESS INVESTIGATION HepG2 cells by the oxidative stress in monoculture and OF AN L-AMINOACID OXIDASE ISOLATED in co-culture. Therefore, BjussuLAAO-II represents a FROM BOTHROPS JARARACUSSU IN AN IN promising compound in the development of new VITRO MODEL USING A CO-CULTURE antitumor drugs. INCLUDING HUVEC AND HEPG2 HUMAN Financial support / Acknowledgments: CAPES, CELLS FAPESP (Proc. Nº 2011/23236-4).

MACHADO, A.R.T.1; AISSA, A.F.1; RIBEIRO, D.L.2; HERNANDES, L.C.1; SAMPAIO, S.V.1; TO 02 - IN VITRO NEPHROTOXICITY OF ANTUNES, L.M.G.1 LAAO ISOLATED FROM CROTALUS ATROX VENOM 1 Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto - SP, Morais, I.C.O. 1; Araújo, D.D.C. 1; Cunha, L.S. 1, da Brasil; 2 Faculdade de Medicina de Ribeirão Preto, Silva, E 1; Orzaez, M 2. Universidade de São Paulo, Ribeirão Preto - SP, Brasil. 1 Centro Universitário Católica de Quixada- UNICATOLICA, Quixada-CE, Brasil; 2. Centro de Hepatocellular carcinoma is one of the most common Investigacion Príncipe Felipe- CIPF, Valencia, types of cancers in adults whose rate increases Espanha. worldwide. Transplantation is the best choice of treatment, but many difficulties leads chemotherapy to Introduction: In Brazil, snake envenomation is a well- be widely used. Chemoresistance and many adverse known cause of death and morbidity. Crotalus venom effects can occur which improves the search for new consists mainly of proteins and peptides, such as L- therapeutic molecules. L-amino acid oxidases (LAAO) amino acid oxidase (LAAO), convulxine, gyroxine, isolated from snake venoms have demonstrated crotoxin and crotamine. Acute Renal Failure (ARF) is cytotoxicity in tumor cell lines. These results represent a common complication caused by Crotalus snakebite in vitro monoculture systems. Recent studies have with relevant morbidity and mortality. Pathogenesis of shown that the tumor microenvironment plays a ARF may be related to hypovolemia and significant role in chemotherapeutic resistance, hypoperfusion secondary to cardiovascular neoplastic transformation, tumor growth, and invasion. disturbances, deposit of fibrin in the glomerular A LAAO purified from Bothrops jararacussu venom capillaries leading thrombotic microangiopathy and (BjussuLAAO-II) was evaluated in hepatocellular high venom concentration at the renal tissue, direct carcinoma cells (HepG2) in monoculture and co- venom action on the tubular cells. Aim: To elucidate culture with human umbilical vein endothelial cells the direct nefrotoxicity of the L-aminoacid oxidase (HUVEC) to simulating the tumor microenvironment. isolated by Crotalus Atrox (LAAO-Atx) venom by Cytotoxic activity was assessed by the MTT and using in the human proximal tubule epithelial cell line clonogenic survival assays, genotoxic activity by (HK-2 cells). RESULTS: In these cells treated with comet assay, oxidative stress by fluorescence, and LAAO Atx, 0.78 – 100 µg/mL for 12 h, there was a chromosome damage by the micronucleus assay. The decrease in their viability in a concentration-dependent results obtained for HepG2 in monoculture showed manner. We next evaluated if necrosis was implicated cytotoxicity and increase in oxidative stress in all in the cellular viability decrease observed by analyzing concentrations tested (0.25 - 5.00 μg/mL). The lactate dehydrogenase (LDH) release. LDH release concentration of 5.00 μg/mL induced DNA, but not was observed after 12 h of LAAO Atx exposure at the chromosomal damage. In co-culture with HUVEC highest concentrations studied when compared with cells, 1.00 and 5.00 μg/mL of BjussuLAAO-II were untreated cells. Annexin V/PI staining was applied to cytotoxic and increased levels of reactive oxygen detect apoptotic/necrotic cells after LAAO Atx species in HepG2 cells; 5.00 μg/mL induced DNA treatment. In accordance with data obtained in the damage. The DNA damage can be correlated to the LDH-release assay, the Annexin-V-PI loading cell increase of intracellular reactive oxygen species. The analysis demonstrated an increase in necrotic (PI+ differences in results between mono and co-culture cells) and secondary necrotic cells (Annexin-V+, PI+) may vary due to the simulation of part of the tumor in a concentration-dependent manner. The ability of microenvironment. In HUVEC monoculture, all LAAO to induce apoptosis in Hk2 cells was further concentrations tested were cytotoxic and genotoxic at demonstrated using caspase-3/7 activity assays. concentrations of 1.00 and 5.00 μg/mL. No LAAO- Atx induced apoptosis at the highest concentrations tested could induce chromosomal concentration studied (50 µg/mL). CONCLUSIONS: damage. In conclusion, the tumor microenvironment Our results suggesting that LAAO from Crotalus Atrox influences the cellular response. Moreover, ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 217 causes nephrotoxicity by acting in multiple cell death underdeveloped countries, affecting remote rural areas pathways. with scarce and underestimated epidemiological data. Financial support / Acknowledgments: CAPES. Financial support / Acknowledgments: INSTITUTO DR. JOSÉ FROTA, UNIVERSIDADE FEDERAL DO CEARÁ, UNIVERSIDADE DE FORTALEZA. TO 03 - SNAKEBITE BY THE BUSHMASTER (Lachesis muta) IN PACOTI-CEARÁ: A CASE REPORT TO 04 - ACUTE PANCREATITIS DUE TO SCORPIONISM MAGALHÃES, K. N.; VERAS, M.S.B.; ALBUQUERQUE, P.L.M.M.; HOLANDA, F.M.T.; MAGALHÃES, K. N.; ALBUQUERQUE, P.L.M.; MOURA, C.T.M.; CUNHA, B.L.; SILVA, SILVA, MOURA, C.T.; PAIVA, J.H.H.G.L.; VERAS, M.S.B., J.F.; SALES, T. C.; GRANJEIRO, A.K.P. HOLANDA, F.M.T.; FREITAS, R.M.; BRIZENO, M.O.; LUSTOSA, F.M. INSTITUTO DR. JOSÉ FROTA INSITUTO DR. JOSÉ FROTA, UNIVERSIDADE INTRODUCTION: In Brazil, there are 70 species of ESTADUAL DO CEARÁ, UNIVERSIDADE DE venomous snakes, most of them genus Bothrops, FORTALEZA Crotalus, Micrurus and Lachesis, which account for about 22,000 snakebites per year. The genus Lachesis INTRODUCTION: Scorpion accidents are an accounts for only 1.4% of ophidian accidents in Brazil. important public health problem due to the high It is known that the L. muta venom has the following frequency of their occurrence and their potential biological activities: proteolytic, coagulant, severity. They usually present a benign course, hemorrhagic and neurotoxic (vagomimetic). although there are reports of serious complications. OBJECTIVE: To report a case of snakebite accident OBJECTIVE: The aim of this study is to describe a by L. muta. CASE REPORT: Patient male, 29, from severe clinical case due to scorpion sting. CASE Pacoti-Ceará, gives entry to the emergency with a REPORT: A 69-year old female, with diabetes mellitus history of stinging by "surucucu" on the left leg 10 and systemic arterial hypertension, was admitted to the hours ago. Patient evolved with intense vomiting, emergency room 7 hours after scorpion injury in right bleeding at the site of the bite, pain and edema that hand. The patient referred intense and diffuse quickly ascended through every affected limb. Initially abdominal pain, sweating and one episode of emesis. he presented bradycardia (HR:52 bpm), hypotension The physical examination presented a lower level of (PA:90x60 mmHg), lethargy, abdominal cramps and consciousness, normal cardiac and pulmonary diarrhea, sweating, blurred vision, prostration. At the auscultation, painful abdomen on superficial palpation, site of the bite there was important edema and systolic pressure of 150mmHg and diastolic pressure hematoma, in addition to bleeding at the venom of 110 mmHg. The admission exams revealed: glucose inoculation sites. Premedication with promethazine = 274mg/dl, amylase = 1914UI/l, lipase = 5442UI/l, 25mg (IM) and hydrocortisone 200mg (EV) was LDH = 474U/l, TGO = 85U/l, TGP = 93U/l, lactate = carried out followed by infusion of 15 ampoules of 2.5mmol/l and potassium = 3.1mEq/l. The antilaquético/antibotrope serum available at the center. electrocardiogram showed signs of hypokalemia, such Submitted 2 hours after the serotherapy, fasciotomy as prominent U waves after T waves. An urgency and tetanus prophylaxis. Sixteen days after the surgical ultrasonography was also performed, which presented procedure the lesion continues under plastic surgery moderate hepatic steatosis. Therefore, the hypothesis surveillance and specialized stomatherapy assistance of acute pancreatitis secondary to scorpianism was The normalization of the coagulogram in 24 hours much more probably. Thus, it was decided to prescribe after the serotherapy. Evolution failure (Urea 104 mg / vigorous intravenous hydration, zero diet, 10% dL and creatinine 2.5 mg / dL and was recovery of the potassium chloride in an attempt to control renal function in 5 days. DISCUSSION/ hypokalemia and anti-scorpion serum (ASS) - 6 CONCLUSION: Although the lachético venom ampoules. After using ASS, the patient reported presents biological activities similar to those of the improvement of abdominal pain, with amylase and genus Bothrops, the signs and symptoms are little lipase values in increase curve during hospitalization. known by the scarcity of cases reported in the However, the patient evolved with acute kidney injury Brazilian literature. The time between the sting and the (urea = 115 and creatinine = 2.4), being chosen beginning of the serotherapy may be determinant for conservative treatment, with a rapid improvement of the occurrence of complications such as compartment renal markers. After 11 days of hospitalization and syndrome, necrosis and acute renal failure. clinic stabilization, the patient was discharged from the Unfortunately, many ophidian accidents occur in hospital. Recently, the patient returned to the nephrology ambulatory, 4 months after of the accident ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 218 and presented complete recovery of kidney and pancreatic function (29/06/2017: Cr = 0,8mg/dl, Ur= 42mg/dl, amylase= 70U/l; lipase= 65mmol/l). CONCLUSION: Acute pancreatitis secondary to scorpionism is extremely rare. Financial support / Acknowledgments: PREFEITURA MUNICIPAL DE FORTALEZA.

VETERINARY TOXICOLOGY ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 220

VT 01 - ANIMAL INTOXICATION: ANALYSIS VT 02 - RESEARCH ON AGROCHEMICALS IN OF THE CASES REGISTERED IN THE ROAD-KILL GIANT ANTEATERS IN THE DATATOX SYSTEM OF ONE CIATOX STATE MATO GROSSO DO SUL

DELGOBO, C. L. 1,2; TAKAHASHI, C. Y. 1; YOGUI, D. R. 1,2; ALVES, M. H. 1,3; DESBIEZ, A. WEBER, J. F. 1; SEEMANN, T. M. S. 1; ZANNIN, J. L. 1,4 M. 1,3 1. Instituto de Conservação de Animais Silvestres, 1 Centro de Informações Toxicológicas de Santa Campo Grande - MS, Brasil; 2. Nashville Zoo, Catarina–CIT/SC/SUR/SES e HU/UFSC; 2 Depto de Nashville - TN, USA; 3. Houston Zoo, Houston - TX, Análises Clínicas - ACL/CCS/UFSC - USA; 4. The Royal Zoological Society of Scotland, Florianópolis/SC; 3 Depto de Patologia e Toxicologia– Edinburgh, U.K. PTL/CCS/UFSC–Florianópolis/SC. Introduction: The Giant anteater (Myrmecophaga Introduction: Poisoning in animals are increasingly tridactyla) is an iconic South American mammal that is frequent in routine service in veterinary medicine, with listed as “Vulnerable” by the IUCN. This animal is prevalence of accidental cases; however, the found in the Brazilian Cerrado which is considered as intentional poisoning by extermination attempts was one of the species strongholds. However, in the last 35 also registered. Objective: To describe the toxic animal years, more than half of Brazil’s Cerrado has been accidents registered in the CIATox of reference in the converted into pasture or agricultural lands for cash region. Material and methods: Retrospective study of crops that use agrochemicals to eliminate ants, termites toxic accident records on animals registered in the first and other pests to increase productivity. Furthermore, version of the DATATOX System of a CIATox in the there is an extensive road network that inflicts a period from 2014 to 2016. Results: In the studied significant mortality rate on local wildlife. Objective: period were recorded 360 cases of accidents in The goal of this study is to collect samples of road-kill animals. Among those, 301 (83.6%) were in canines, Giant anteaters, to identify and quantify the 32 (9%) in cats, 15 (4.2%) in cattle, six (1.7%) in agrochemicals that are absorbed by them and, if horses, three (0.84%) in birds, one (0.56%) in wild possible, relate these results with microscopic and animals, and two with indefinite family. The classes of macroscopic findings. Material and methods: A team agents involved were: in canines, pesticides, with of veterinarians will drive 1.337 km on the highways 34.4%, followed by venomous animals 22.3%, of the state Mato Grosso do Sul, every two weeks and rodenticides 8%, veterinary products 6%, plants 6%, at approximately 60 km/h, searching for Giant anteater industrial chemical products 6%, medicines 5.6%, casualties. The following biological material will be sanitizing 2.6%. In cats: pesticides, venomous animals collected: nostril, paw pad and bone, packed in zip- and veterinary products with 16% and medicines with closed plastic bags. If the carcass is fresh, a necropsy 10%. In equines: medicines, pesticides and venomous will be performed and material from liver, spleen, animals. In birds: venomous animals (bees) and kidney, fat and blood clot will be collected in Falcon veterinary products. In wild animals: pesticides. 360 tubes. All these samples will be stored in a freezer at – cases were analyzed, and among those, 39 cases 20 °C. Partnerships will be established with (10.8%) evolved to death, being 35 for intoxication. laboratories to analyze the samples, looking for Concerning the poisoning deaths, 23 were canines, organophosphates, organochlorines, pyrethroids, being 17 by venomous animals (coral, jararaca and carbamates and other substances of interest. Results: bee), 3 for pesticides (carbamate and phosphorous), From February to July 2017, 16.659 km were surveyed one by rat poison (coumaric), one for medicine and 143 road-kill Giant anteater have been counted. (biguanidas) and one by plant; 10 cattle (sulphur in the Necropsies of 20 Giant anteaters were performed and food); one cat (indeterminate) and one bird (antibiotic). the following biological material has been collected: Conclusion: the largest number of animal poisoning 31 samples of paw pads, 31 of nostril, 30 of bone, 8 of recorded in CIATox were in domestic animals: canines spleen, 7 of liver, 17 of kidney, 2 of fat and 5 of blood followed by cats. The class of agent mostly involved clot. In this phase of the study we are looking for were the pesticides and the largest number of deaths partner laboratories to analyze the samples. was in canines by venomous animals. Conclusion: After 6 months of monitoring, data demonstrate that Giant anteaters are the third most Financial support / Acknowledgments: Centro de killed animals on the surveyed highways. It is possible Informações Toxicológicas de Santa Catarina– to collect biological materials and the next step is find CIT/SC/SUR/SES e HU/UFSC. partnerships to analyze the samples.

ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 221

Financial support / Acknowledgments: Anteaters histological data are still under analysis. Therefore, the and Highways Project is a partnership between partial data allow to verify that the used NSAIDs Fondation Segre, Zoo Conservation Outreach Group, promoted a lower weight gain and a lower intake of The Royal Zoological Society of Scotland and Instituto food by the birds, however, could these changes be due de Conservação de Animais Silvestres, all project to the palatability of the salicylates added in the feed? donors are listed at Biochemistry and histology will greatly contribute to http://www.giantanteater.org/supporters.html. answering this question. Financial support / Acknowledgments: This work was supported by the Coordenação de VT 03 - COMPARATIVE STUDY OF Aperfeiçoamento de Pessoal de Nível Superior, ACETYLSALICYLIC ACID, SALICYLIC ACID CAPES, Brazil [grant number 1723701/2017]. AND SODIUM SALICYLATE IN BROILER CHICKENS VT 04 - ANALYTICAL DETECTION OF HEAVY ALMEIDA, E. R. M. 1; GÓRNIAK, S. L. 1; METALS AND PESTICIDES IN CARCASSES OF ANDRÉO-FILHO, N. 2; ARAUJO, C. S. S. 2; WILD LOWLAND TAPIRS (Tapirus terrestris) GOTARDO, A. T. 1; HUEZA, I. M. 1,2. FROM ROAD-KILL IN THE STATE OF MATO GROSSO DO SUL, BRAZIL 1. Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP), São Paulo - SANTOS, R. C. F. 1,2,3; TESTA, C. 1,2; MEDICI, E. SP, Brasil; 2. Universidade Federal de São Paulo P. 1,2,4 (UNIFESP), Diadema - SP, Brasil. 1 Iniciativa Nacional para a Conservação da Anta Salicylates are non-steroidal anti-inflammatory drugs Brasileira (INCAB), Instituto de Pesquisas Ecológicas (NSAIDs) with analgesic and antipyretic properties. (IPÊ), Nazaré Paulista - SP, Brasil; 2 International They are generally quite safe and commonly used in Union for Conservation of Nature (IUCN) Species veterinary medicine in most mammalian species and Survival Commission (SSC) Tapir Specialist Group promote adverse effects that can lead to the suspension (TSG), Campo Grande - MS, Brasil; 3 Instituto of their use and also therapy failure, depending on the Brasileiro para Medicina da Conservação (TRÍADE), dose and the period of administration. However, the Curitiba - PR, Brasil; 4 Escola Superior de knowledge about their effects in broilers chicken when Conservação Ambiental e Sustentabilidade given for a prolonged period using extrapolated (ESCAS/IPÊ), Nazaré Paulista - SP, Brasil. therapeutic doses from wild birds is still unknown. Therefore, the objective of this study was to compare There are very little information on wildlife the safety of the use of salicylates: acetylsalicylic acid ecotoxicology and on pesticide effects on the (ASA), salicylic acid (SA) and sodium salicylate (SS), physiology and ecology of wildlife species. The when administered in small doses daily, and their objective of this study was to investigate the presence impact on growth and feed consumption of broiler of heavy metals and pesticides in biological samples of chickens. A total of 96 male Cobb 500 chickens were wild lowland tapirs (Tapirus terrestris) from road-kill divided in four groups with two repetitions (one in the State of Mato Grosso do Sul, Brazil. The study control group – CO; and three treatments) into 1.0 m x was carried out in four highways (BR-262, BR-267, 1.0 m boxes with 12 birds each. Drugs were given at a MS-040, and MS-145) that cross areas of Cerrado dose of 5 mg/kg added to feed per day, during 21 days. fragments, pasture lands, agricultural crops Feed and water were supplied ad libidum. Weekly, the (particularly sugar cane, soybean and corn), and rural birds weight and food consumption were measured. On communities, between September 2015 and November experimental day 21 , blood was collected for 2016. Fresh carcasses of 25 wild lowland tapirs were hemogram hemogram and biochemistry analysis , as submitted to necropsy, and biological samples well as tissue for histology. As a partial results, it was collected were stored frozen until the analysis by the observed a significant increase in the number of Toxicological Assistance Center (CEATOX) of lymphocytes from the SA group and a decrease in the Botucatu, in São Paulo State. The laboratory number of eosinophils from SS group when compared evaluation included the following techniques: (a) with the others groups. Regarding to birds’ weight , it atomic absorption spectrophotometry (AAS) to detect was observed a decrease in the weight gain and the heavy metals; (b) gas chromatography - electron feed consumption of the birds of all the groups treated capture detector (GC-ECD) to detect organochlorines, with NSAIDs, from the second week of treatment organophosphates, and pyrethroids; and (c) high when compared with the CO group. In addition, the SS performance liquid chromatography - ultraviolet group obtained the lowest reduction in this parameter. (HPLC-UV) to detect carbamates. A total of 132 Until the present moment, the biochemical and biological samples were analyzed, including: bones ISSN: 2359-4721 Volume 2 / Suppl. 2 / 2017 222

(17), nail (22), footpad (24), proboscis (5), stomach content (25), intra-cardiac blood (14) and liver fragments (25). Twelve different compounds were detected in 67 percent of the samples evaluated in this study (88/132), including cadmium (0,30±0,23µg/g-1), lead (1,21±0,58µg/g-1), copper (3,69±12µg/g-1), manganese (1,82±1,37µg/g-1), diazinon (47,48±11,94ppb), mevinphos (185,90±164,05ppb), malathion (306,57±208,25ppb), dimethoate (26,70±14,47ppb), aldicarb (5,32±15,17µg/g), carbaryl (0,11±0µg/g), deltamethrin (0,47±0,36µg/g), and permethrin (0,43±0,24µg/g). Macroscopic alterations of the liver and/or kidneys were observed in 88 percent of the individuals (22/25), and the top differential is currently the chronic over-exposure to pesticides. This study is part of a complete health assessment of wild lowland tapirs in the Brazilian Cerrado, and investigating the effect of pesticides on tapir health is a core goal of this project.

Financial support / Acknowledgments: Tapir health assessments are a component of the long-term activities of the Lowland Tapir Conservation Initiative (LTCI) carried out by the Instituto de Pesquisas Ecológicas (IPÊ) in Brazil. The LTCI has the institutional support from the International Union for Conservation of Nature (IUCN) Species Survival Commission (SSC) Tapir Specialist Group (TSG); Association of Zoos and Aquariums (AZA) Tapir Taxon Advisory Group (TAG); European Association of Zoos and Aquariums (EAZA) Tapir Taxon Advisory Group (TAG); and several zoos and institutions worldwide. To this project, the authors would like to thank the Foundation Segré and the Whitley Fund for Nature for the financial support, and the Toxicological Assistance Center (CEATOX) of Botucatu for the partnership for the laboratory analyzes.