IJUP08 First Meeting of Young Researchers of U.Porto 20•22 Feb 2008

Faculdade de Arquitectura da Universidade do Porto

RESUMOS / ABSTRACTS

Nota Introdutória

A Universidade do Porto inicia em 2008, sob o tema "Investigação Jovem na Universidade do Porto (IJUP)", uma série de encontros científicos destinados aos seus alunos (nacionais ou de Eramus) dos 1º e 2º ciclos. O IJUP é um encontro científico, de cariz anual, onde os alunos da U. Porto são convidados a apresentar trabalhos ou estudos por eles desenvolvidos ou em vias de serem desenvolvidos, enquanto estudantes do 1º ou 2º ciclos. São objectivos principais do IJUP fomentar a participação dos alunos da U. Porto em eventos de índole científica, permitir que os seus trabalhos sejam divulgados junto da comunidade científica da U. Porto e dos restantes alunos desta Universidade e incentivar a discussão interdisciplinar entre as diferentes vertentes de investigação que são desenvolvidas na U. Porto. A comissão organizadora do IJUP é composta por elementos de todas as unidades orgânicas da U. Porto e tem a coordenação do Vice-Reitor da U. Porto para ID&I. No IJUP08, a vertente interdisciplinar será reforçada e francamente enriquecida pela participação de várias instituições internacionais convidadas, como são o caso das universidades brasileiras: Universidade de São Paulo (USP) e Universidade Estadual Paulista (UNESP) e norte americanas: The University of Texas at Austin e da Rutgers, The State University of New Jersey. Contando com uma enorme diversidade de áreas do conhecimento, o IJUP08 tem prevista a realização de duas sessões plenárias de comunicações orais interdisciplinares que têm como objectivos promover o contacto e a comunicação entre áreas tão diferentes como as ciências da saúde, a arquitectura, a psicologia, a engenharia, a economia, entre outras. Fazem também parte do programa do IJUP 08 várias sessões de comunicações orais que ocorrerão em paralelo e que estão agrupadas consoante a área científica em que se inserem, e ainda sessões de apresentação de posters em que se prevê, mais uma vez, a discussão interdisciplinar promovida por pequenas comissões de moderadores de diferentes áreas científicas. Os objectivos científicos, pedagógicos e de divulgação interdisciplinar, do IJUP08 serão consolidados pela realização de um workshop que pretende a divulgação de diversas oportunidades de financiamento disponibilizadas em Portugal e de um outro que permitirá o contacto dos jovens investigadores com investigadores seniores da U.Porto com carreira reconhecida, de diferentes áreas científicas e diferentes gerações. Pela afluência, diversidade e qualidade das participações, esperamos que o IJUP08 cumpra com sucesso todos os seus objectivos, motivando assim a comunidade académica à continuação e incentivo da Investigação Jovem.

A Comissão Organizadora

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Comissão Organizadora / Comité Científico

Adrián Silva Dinis Cayolla Ribeiro Elisa Keating Filipe Abrantes Georgina Correia da Silva Gonçalo Furtado Helena Pereira Madureira Hugo Valente Jorge Gonçalves Jorge Teixeira José Rodrigues Lima Laura Oliveira Lígia Afonso Luís Miguel Palma Madeira Mª Paula Santos Manuela Aguiar Marcela Alves Segundo Maria José Oliveira Pedro Sousa Gomes Sofia Rodrigues

Secretariado: Helena Oliveira Maria João Amaral

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Índice

NOTA INTRODUTÓRIA ______I

COMISSÃO ORGANIZADORA / COMITÉ CIENTÍFICO______II

PROGRAMA GERAL______1

INTERDISCIPLINARY ORAL SESSIONS - IOS1 ______4

WHAT CAN A SIMULATION STUDY TELL US ABOUT THE RELEVANCE OF UTILITY AND WEALTH INDICATORS FOR DECISION MAKING CONCERNING PRODUCTION AND MARKET BEHAVIOUR______5

CONSTRUCTION OF A LEARNING UNIT IN A MOODLE PLATFORM: STUDYING MITOSIS USING FRUIT FLY AS A MODEL ______6

THE KARIMOJONG PEOPLE FROM UGANDA: A HISTORY ASSESSED THROUGH PATTERNS OF MITOCHONDRIAL DNA ______7

ANGIOGENESIS MODULATION BY CATECHIN ______8

NOVEL FUNCTIONALISED TEXTILES THROUGH NANOSTRUCUTURED CARBON AND CLAY BASED MATERIALS (NANOTEXTUP) ______9

ALFONSO X’S GENERAL ESTORIA AND THE PORTUGUESE CULTURE: AN EXAMPLE OF CONTINUITY BETWEEN MIDDLE AGES AND MODERNITY ______10

INTERDISCIPLINARY ORAL SESSIONS – IOS2______11

ALTERNATIVE USER INTERFACES FOR GAME CONTROL______12

TRADITIONAL TIMBER CONSTRUCTION BUILDINGS: THE EXAMPLE OF THE PORTUGUESE CENTRAL LITTORAL PALHEIROS ______13

TREATMENT OF MYASTHENIA GRAVIS WITH CHOLINESTERASE INHIBITORS: ON THE ROLE OF NICOTINIC AUTORECEPTORS CONTAINING Α3Β2 SUBUNITS ____ 14

MUSIC AND PHONOLOGICAL AWARENESS______15

AUTOMATIC EMOTION ASSESSMENT THROUGH BIOMETRIC INFORMATION ___ 16

DEVELOPMENT OF MOLECULAR BIOLOGY TECHNIQUES FOR THE DETECTION OF GENETICALLY MODIFIED ORGANISMS IN MAIZE FOOD PRODUCTS ______17

PERMEABILITY CHARACTERISTICS OF HIGH PERFORMANCE CONCRETE (HPC) CONTAINING VARYING CONCENTRATIONS OF HYCRETE TESTED AND COMPARED BY METHOD OF RAPID CHLORIDE PERMEABILITY TEST AND THE NON-STEADY-STATE MIGRATION TEST (NORDTEST METHOD NT BUILD 492)___ 18

MARIO PEDROSA E A SUA ATUAÇÃO CRÍTICA NA PRESIDÊNCIA DA ABCA _____ 19

PARALLEL ORAL SESSIONS I – POS.I.1 ______20

YEAST AS A NEW APPROACH TO STUDY THE REGULATION OF P53-DEPENDENT APOPTOSIS BY PROTEIN KINASE C DELTA______21

ANTITUMOR AND ANTIFUNGAL ACTIVITIES OF XANTHONIC DERIVATIVES, POTENTIAL INHIBITORS OF STEROID SULFATASE ______22

E-CADHERIN SILENCING: MORE THAN INACTIVATING MUTATIONS ______23

SYNTHESIS AND DETERMINATION OF PHYSICOCHEMICAL PARAMETERS OF XANTHONES ______24

DISSEMINATION OF BLATEM-52 AMONG ESCHERICHIA COLI AND KLEBSIELLA PNEUMONIAE STRAINS IS CAUSED BY SPREAD OF PLASMIDS IN PORTUGAL __ 25

SYNERGISM BETWEEN ADENOSINE A2A AND TACHYKININ NK1 RECEPTORS FACILITATE [3H]-ACH RELEASE FROM MYENTERIC NEURONS ______26

PARALLEL ORAL SESSIONS I – POS.I.2 ______27

THE KNOWLEDGE-BASED ECONOMY AS A HEGEMONIC META-OBJECT OF URBAN CENTRE RENEWAL POLICIES: THE CASE OF PORTO ______28

“MASSARELOS NETWORK”: SPACES AND TIMES OF INTER-INSTITUTIONAL ARTICULATION, IN THE PERSPECTIVE OF AN “EDUCATING PARISH” ______29

THE SPACE OF INTERVALS. THE INNER SPACES OF URBAN BLOCKS: SPACE, LANDSCAPE, CITY______30

FORMS OF HOUSING AND WAYS OF LIVING IN PORTO’S CONTEMPORARY URBAN TERRITORY ______31

INTERACTIVE APPLICATION FOR SPATIAL COMMUNICATION AND REPRESENTATION ______32

ECONOMICAL AND SOCIAL BENEFITS OF ASTRONOMY ______33

PARALLEL ORAL SESSIONS I – POS.I.3 ______34

ESTIMATION OF POSTMORTEM INTERVAL BASED ON THE DETERMINATION OF POTASSIUM IN VITREOUS HUMOR BY AN AUTOMATIC SYSTEM ______35

THE DYNAMICAL STATE OF GALAXIES: FROM 2D TO 6D ______36

DEVELOPMENT OF AN AUTOMATIC MULTI-PUMPING FLOW SYSTEM FOR THE SPECTROPHOTOMETRIC DETERMINATION OF TRIMIPRAMINE______37

ANTI-DE SITTER SPACE AND THE STABILITY OF SCALAR FIELD ______38

ACCESSIBILITY AND SCIENCE COMMUNICATION – NEW DIRECTIONS AND GUIDELINES FOR MOODLE “ESTALEIRO DA CIÊNCIA”______39

QUANTUM VACUUM ENERGY IN GENERAL RELATIVITY (THE CASIMIR EFFECT) 40

PARALLEL ORAL SESSIONS II – POS.II.1 ______41

MICROPLATE HIGH-THROUGHPUT METHODOLOGY FOR DETERMINATION OF FOLIN-CIOCALTEU INDEX ______42

DETECTION OF AUTHORIZED GENETICALLY MODIFIED MAIZE EVENTS: PARTICIPATION IN AN INTER-LABORATORIAL STUDY______43

DETECTION OF GENETICALLY MODIFIED SOYBEAN IN FOODSTUFFS: PARTICIPATION IN AN INTER-LABORATORIAL STUDY______44

HPLC SEPARATION AND QUANTIFICATION OF CATECHINS IN GREEN TEA HOME PREPARED: COMPARISON WITH COMMERCIAL SOFT DRINKS ______45

PRODUCT ENGINEERING – APPLICATION OF VANILLIN IN ALIMENTARY FOOD PASTE INDUSTRY ______46

DEVELOPMENT OF A COATING BARRIER FOR CORK STOPPERS FOR THEIR USE IN SPIRITUOUS DRINKS ______47

PARALLEL ORAL SESSIONS II – POS.II.2 ______48

THE ARCHITECT’S HOUSE ______49

ORGANIC LIVING | WEEKEND HOUSE IN CASTELO DE PAIVA ______50

FROM RCR EXPERIENCE TO A PROJECT OF A SMALL HOUSE IN THE MOUNTAIN ______51

CASA COMUM [COMMON/COLLECTIVE/COMMUNITARIAN HOUSE/HOME] ARCHITECTURES FOR AN INTERMEDIARY SPACE ______52

THE COURTYARD AND SIZA’S HOUSES______53

THE SILENCE AND THE SKY: SECLUSION AND ARCHITECTURE. FOUR COURTYARD-HOUSES ______54

PARALLEL ORAL SESSIONS II – POS.II.3 ______55

EFFECT OF METALS ON THE ATPREP1 PEPTIDASOME ACTIVITY ______56

LIPOSOME-INCORPORATED ZANTHOXYLUM TINGOASSUIBA ESSENTIAL OIL: PREPARATION AND CHARACTERIZATION______57

ANCHORING OF MACROCYCLE COMPOUNDS ONTO NANOSTRUCTURED CARBON MATERIALS FOR CATALYTIC APPLICATIONS ______58

CHARACTERIZING THE INTERMETALLIC FORMED DURING BALL ATTACH PROCESS ______59

FUNCTIONAL INSULIN QUANTIFICATION UPON ITS NANOENCAPSULATION ____ 60

TEMPERATURE AND COMPOSITION EFFECTS ON THE MICELLE FORMATION BY A CATANIONIC SURFACTANT: A SURFACE TENSION AND CONDUCTIVITY STUDY 61

PARALLEL ORAL SESSIONS III – POS.III.1 ______62

PHENOLIC COMPOUNDS, ORGANIC ACIDS AND ANTIOXIDANT PROPERTIES OF RUMEX INDURATUS ______63

MUC1 OVEREXPRESSION IS ASSOCIATED WITH DISTANT METASTASES DEVELOPMENT IN CANINE MAMMARY CARCINOMAS ______64

ANALYSIS OF THE EFFICIENCY OF THE MITOTIC CHECKPOINTS IN GLIOBLASTOMA CELL LINES ______65

PRENYLATED AND BROMOALKYLATED XANTHONES AS POTENTIAL ANTITUMOR AGENTS: SYNTHESIS AND BIOLOGICAL ACTIVITIES ______66

P-CADHERIN: ROLE IN BREAST CANCER CELL MIGRATION AND INVASION ____ 67

IMPAIRMENT OF RETROGRADE SIGNALLING VIA ADENOSINE IN TOXIN-INDUCED MYASTHENIA GRAVIS: CROSS-TALK WITH MUSCARINIC AUTORECEPTORS ___ 68

PARALLEL ORAL SESSIONS III – POS.III.2 ______69

CONTEMPORARY CONDITION: ‘NEW’ RELATIONSHIPS BETWEEN ARCHITECTURE AND CITY ______70

FORM AND MATTER IN THE WORK OF RAFAEL MONEO ______71

ARCHITECTURE AND CLIMATE. SITE GEOGRAPHY: LUANDA AND THE WORK OF VASCO VIEIRA DA COSTA ______72

CRISES E REFORMAS: O COLONIALISMO ILUSTRADO NA CAPITANIA DE SÃO PAULOE OS ESFORÇOS DE MODERNIZAÇÃO DO GOVERNO MORGADO DE MATEUS (1765-1755) ______73

INCORPORATION OF ORGANICS/INORGANICS NANOCOMPOSITES IN COATINGS OF HIGH ENVIRONMENTAL AND MECHANICAL RESISTANCE______74

ZENITHAL ILLUMINATION – FROM ALVAR AALTO TO ÁLVARO SIZA’S WORK ____ 75

PARALLEL ORAL SESSIONS III – POS.III.3 ______76

ARCHITECTURAL LANGUAGES FOR THE 21ST CENTURY ______77

EXHIBITING ARCHITECTURE IN PORTUGUESE. CURATORIAL ACTIVITY IN PORTUGAL PRE- TRIENAL DE ARQUITECTURA DE LISBOA ______78

THE “ECLIPSE” OF ARCHITECTURE ______79

PROMENADE TO CORBUSIER ______80

ARCHITECTURE: STUDIES AND POSSIBILITIES OF TRANSVERSALITIES. SPACE OF ENCOUNTER – HERZOG & DE MEURON, DILLER + SCOFIDIO, DAN GRAHAM, LACATON & VASSAL, SANCHO Y MADRIDEJOS, VITO ACCONCI [1] ______81

CONTACT POINTS_BETWEEN SWISS AND PORTUGUESE ARCHITECTURE_____ 82

PARALLEL ORAL SESSIONS IV – POS.IV.1 ______83

LEVELS OF BUTYLTINS IN SEDIMENT AND SUITABILITY OF SALT MARCH PLANTS FOR RHIZOREMEDIATION ______84

IMPLEMENTATION OF AUTOMATIC METHODOLOGIES FOR QUALITY CONTROL OF PHARMACEUTICAL FORMULATIONS CONTAINING ALLOPURINOL BASED ON BIOCATALYSTS ______85

AZO-DYE ORANGE II DEGRADATION BY FENTON-LIKE REACTION USING AS CATALYSTS TRANSITION METALS IN CARBON SUPPORTS ______86

MONITORING OF BIOLOGICAL REMEDIATION OF A SOIL POLLUTED WITH PETROLEUM HYDROCARBONS______87

ISOLATION AND CHARACTERISATION OF A NOVEL PLANCTOMYCETE BACTERIUM RELATED TO ISOSPHAERA GROUP FROM A FRESHWATER AQUARIUM ______88

INTRACELLULAR GENE REGULATION VIA ENGINEERED NANOLIPOBLOCKERS FOR INHIBITION OF CHOLESTEROL UPTAKE ______89

PARALLEL ORAL SESSIONS IV – POS.IV.2 ______90

THE RELATIVITY OF HAPPINESS: PORTUGAL AND THE EU ______91

NATURAL RESOURCES COURSE ______92

PROCESSES OF CORRUPTION IN PORTUGAL: RESULTS OF AN EXPLORATORY RESEARCH______93

DESISTÊNCIA DO TRATAMENTO DE ENURESE: AVALIAÇÃO DOS MOTIVOS ALEGADOS E DE VARIÁVEIS ASSOCIADAS ______94

POPULAR CULTURE AND THE FORMATION OF THE WORKING CLASS______95

A TROUBLED LEGACY. CHANGING STRATEGIES OF SOCIAL TRANSMISSION AND PATTERNS OF IDENTITY PRODUCTION AMONG INDUSTRIAL WORKERS ______96

PARALLEL ORAL SESSIONS IV – POS.IV.3 ______97

SCREENING OF PHENOLIC COMPOUNDS WITH BIOACTIVITY IN PIERIS BRASSICAE L. REARED ON BRASSICA RAPA VAR RAPA L. USING HPLC-DAD- MS/MS-ESI ______98

SPATIO-TEMPORAL CHARACTERIZATION OF THE ENDOCANNABINOID SYSTEM ENZYMES-FAAH AND COX-2 IN THE FETOPLACENTAL UNIT ______99

DETERMINATION OF HYPOXANTHINE IN VITREOUS HUMOR BY A SEQUENTIAL INJECTION ANALYSIS SYSTEM AND ITS APPLICATION IN THE ESTIMATION OF POSTMORTEM INTERVAL ______100

A NEW METHOD TO EVALUATE SPONTANEOUS PAIN IN NEUROPATHIC PAIN MODELS: A STUDY IN RATS WITH SPARED NERVE INJURY ______101

COMPREHENSIBILITY OF PICTORIAL REPRESENTATIONS WITHIN HEALTH EDUCATIONAL AND INSTRUCTIONAL ARTIFACTS – A CASE STUDY ______102

CALCIUM UPTAKE BY RAT HIPPOCAMPAL SYNAPTOSOMES: REGULATION BY ENDOGENOUS ADENOSINE AND DIFFERENT DEPOLARIZING AGENTS ______103

NEW CONDITIONS FOR GREATER PURIFICATION YIELD OF PHOSPHORYLATED PERIOD PROTEIN IN DROSOPHILA______104

POSTERS ______105

HPLC-DAD-MS/MS ANALYSIS OF PHENOLICS IN VITRO SHOOTS OF BRASSICA OLERACEA L. VAR. COSTATA DC ______111

THERMODYNAMIC AND THEORETICAL STUDY OF DIPHENYLPYRIDINES _____ 112

FORM AND STRUCTURE ON EDUARDO SOUTO DE MOURA’S ARCHITECTURE 113

POSSIBLE CORRELATION BETWEEN MORBID OBESITY AND INFLAMMATION: A PERIPHERAL LEUKOCYTE STUDY ______114

IN VITRO AND IN VIVO STUDIES OF THE EXPRESSION OF CARBOHYDRATES IN A CANINE MAMMARY CARCINOMA CELL LINE ______115

DETERMINATION OF CADMIUM CONTENT IN SARDINE PILCHARDUS______116

OPTIMIZATION STUDY FOR ARSENIC QUANTIFICATION IN FISH USING MICROWAVE-ASSISTED DIGESTION AND GRAPHITE FURNACE ATOMIC ABSORPTION SPECTROMETRY ______117

OPTIMIZATION OF AN ENZYMATIC AND CHROMATOGRAPHIC METHOD FOR INORGANIC PYROPHOSPHATE BASED ON FIREFLY LUCIFERASE ______118

SURVEY OF THE EFFECTS OF THE ANTIBIOTIC MINOCYCLINE ON FRESHWATER CYANOBACTERIA ______119

LOW-COST MATERIALS AS ADSORBENTS FOR TEXTILE WASTEWATER TREATMENT______120

DETERMINATION OF PHYSICAL PARAMETERS IN PHARMACEUTICAL POWDERS BY NEAR INFRARED SPECTROSCOPY (NIR) ______121

AN EXPEDITIOUS METHOD FOR DETERMINATION OF RESIDUES OF ORGANOCHLORINE PESTICIDES IN SEDIMENTS AND ITS APPLICATION TO PORTUGUESE ESTUARINE AREAS______122

KINETICS OF INHIBITION OF FIREFLY LUCIFERASE BY OXYLUCIFERIN AND DEHYDROLUCIFERYL-ADENYLATE ______123

EFFECT OF PYROPHOSPHATE ON FIREFLY LUCIFERASE BIOLUMINESCENCE 124

MOLECULAR ENERGETICS OF HYDROXYBENZALDEHYDE ISOMERS______125

TOURISTIC TOPOGRAPHIES : TRANSVERSALITY AND AUTHENTICITY IN AN PORTUGUESE CONTEXT ______126

SENSOR FOR AMPEROMETRIC DETERMINATION OF AMMONIA IN SEAWATER 127

COPPER(II)/CIPROFLOXACIN COMPLEXES: SYNTHESIS AND SOLUTION STUDIES ______128

THERMOCHEMICAL STUDIES OF TWO CYCLIC UREAS: IMIDAZOLINONE AND N,N'- TRIMETHYLENEUREA ______129

FEELING ARCHITECTURE. THE IMPORTANCE OF ‘THE OTHER’ SENSES IN THE EXPERIENCE OF SPACE. ______130

DEEP EUTECTIC SOLVENTS: A NEW KIND OF IONIC LIQUIDS ______131

AUTHENTICITY EVALUATION AND MOLECULAR CHARACTERIZATION OF PORTUGUESE VARIETIES OF JUGLANS REGIA USING MICROSATELLITE MARKERS______132

UV PROTECTION OF GEOTEXTILES BY CHIMASSORB 944 ______133

COMPUTATIONAL ANALISYS OF CYNARA CARDUNCULUS L. CARDOSINS POMOTERS ______134

HYDROGEN PEROXIDE REVERSIBLE LUMINESCENCE SENSORS______135

CHROMIUM REMEDIATION USING ZERO VALENT IRON - BATCH STUDIES ____ 136

DEVELOPMENT OF A MULTICOMMUTATED FLOW SYSTEM FOR CHEMILUMINOMETRIC QUANTIFICATION OF GENTAMICIN ______137

POTENTIOMETRIC DETERMINATION OF GENTAMICIN IN PHARMACEUTICAL FORMULATIONS ______138

INFLUENCE OF ORGANIC POLLUTANTS ON PHYTOREMEDIATION OF COPPER BY A SALT MARSH PLANT ______139

DFT STUDY ON THE ABILITY OF CALIX[2]FURANO[2]PYRROLE TO FORM HOST- GUEST COMPLEXES WITH DIFFERENT IONS ______140

FIBER OPTIC PH SENSORS BASED ON CDTE QUANTUM DOTS ______141

STEADY STATE AND LIFETIME CHEMICAL SENSORS BASED ON RUTHENIUM COMPLEXES ______142

MOLECULAR CHARACTERIZATION OF THE PORTUGUESE PATIENTS WITH MUCOPOLYSACCHARIDOSIS IIIC: TWO NOVEL MUTATIONS IN THE HGSNAT GENE ______143

SCOPOLETIN, A NATURAL COUMARIN ISOLATED FROM AGROSTISTACHYS GAUDICHAUDII (EUPHORBIACEAE) ______144

5-(HYDROXYMETHYL)-2-FURFURAL, A NATURAL COMPOUND ISOLATED FROM STROPHIOBLACHIA FIMBRICALYX BOERL. (EUPHORBIACEAE) ______145

NEW APPROACHES FOR ESTIMATING THE POSTMORTEM INTERVAL BASED ON VITREOUS HUMOR TRACE ELEMENT LEVELS ______146

NEW APPROACHES TO THE DIAGNOSTIC OF DEATH BY DROWNING BASED ON TRACE ELEMENT LEVELS IN BLOOD OF THE CARDIAC CAVITIES ______147

FIRING DISTANCE ESTIMATION THROUGH THE ANALYSIS OF THE GUNSHOT RESIDUES DEPOSIT PATTERN AROUND DE BULLET ENTRANCE HOLE ______148

IN VITRO ASSESSMENT OF WHEY EDIBLE COATS WITH ANTIMICROBIAL FEATURES AGAINST DIFFERENTS MICROBIAL STRAINS ______149

SYNTHESIS, STRUCTURE ELUCIDATION AND EVALUATION OF XANTHONE DERIVATIVES FOR DUAL ACTIVITY: ANTITUMOR ACTIVITY AND P-GLYCOPROTEIN INHIBITION ______150

SYNTHESIS OF XANTHONE DERIVATES FOR IN VITRO AND IN VIVO BIOLOGICAL ACTIVITY STUDIES ______151

ULTRASTRUCTURAL CHARACTERISATION OF PIRELLULA SP. OJF20 ______152

MESOPOROUS CARBONS: SYNTHESIS AND FUNCTIONALIZATION______153

PHYSICAL EXERCISE AND STRESS HORMONES ______154

SINGLE-STANDARD CALIBRATION APPLIED TO SIA ANALYTICAL SYSTEM WITH POTENTIOMETRIC DETECTION______155

PHOTOCATALYTIC DEGRADATION OF CIPROFLOXACIN ANTIBIOTIC IN TIO2 AQUEOUS SUSPENSION ______156

ANTI-ANGIOGENIC EFFECTS OF RANIBIZUMAB AND BEVACIZUMAB IN AGE- RELATED MACULAR DEGENERATION: EFFECTS ON HUMAN ENDOTHELIAL CELLS ______157

GEOMETRY AND SPACE CONCEPTION THE IMPORTANCE OF THE PROCESSES OF REPRESENTATION TO ARCHITECTURE______158

WET PEROXIDE OXIDATION AND WET OXIDATION OF NITROPHENOLS IN AQUEOUS STREAMS ______159

FORMATION OF HETEROCYCLIC AROMATIC AMINES IN CHARCOAL GRILLED SARDINES ______160

INFLUENCE OF BEER ANTIOXIDATIVE PROPERTIES ON THE FORMATION OF THE HETEROCYCLIC AMINE PHIP IN GRILLED BEEF ______161

THE STUDY OF DE FÂNZERES-GONDOMAR GRANITES: A MULTIDISCIPLINARY APROACH______162

EDUCATIONAL SCIENCES AND MEDICAL EDUCATION: AN INTERNSHIP IN FCM- UNL DEPARTMENT OF MEDICAL EDUCATION______163

QUARTZ CRYSTAL MICROBALANCE: A USEFUL TOOL AS SENSOR OF THIN FILMS ______164

JOÃO ANDRESEN’S FIFTIES HOUSES: PORTUGUESE DOMESTIC ARCHITECTURE UP-TO-DATE______165

SYNTHESIS OF XANTHONE DERIVATIVES BY MICROWAVE-ASSISTED METHODS ______166

LOCATION AND OWNERSHIP OF THE NATURAL RESOURCES IN IRAQ ______167

SYNTHESIS, STRUCTURAL ELUCIDATION AND BIOLOGICAL ACTIVITIES OF CHIRAL 1-AMINO-2-PROPANOL XANTHONE DERIVATIVES ______168

CHIRAL ALANINOL XANTHONE DERIVATIVES: SYNTHESIS, STRUCTURAL ELUCIDATION AND BIOLOGICAL ACTIVITIES______169

PRIMARY HYPERPARATHYROIDISM: THE UNKNOWN WORLD OF SPORADIC CASES AND THEIR GENETIC ALTERATIONS ______170

FLOW CYTOMETRIC DETECTION OF P-GP MEDIATED DRUG EFFLUX: A METHOD FOR THE EVALUATION OF THE ACTIVITY OF P-GP INHIBITORS ______171

EFFECTS OF ETOPOSIDE, DOXORUBICIN AND CYTARABINE IN BURKITT LYMPHOMA CELL LINES______172

AUTOMATIC FIA METHOD FOR DETERMINATION OF IRON IN ENVIRONMENTAL SAMPLES ______173

CHARACTERIZATION OF EXOTIC WOOD SPECIES FOR FINISHING WITH WATER- BORNE VARNISHES ______174

MICROBIOLOGICAL STUDY OF THE INTERACTION OF OUTER MEMBRANE PROTEINS WITH ANTIBIOTICS______175

INTERACTIONS OF SULINDAC AND ITS METABOLITES WITH PHOSPHOLIPID MEMBRANES: AN EXPLANATION FOR THE PEROXIDATION PROTECTIVE EFFECT OF THE BIOACTIVE METABOLITE ______176

EVALUATION OF PRIMER COATINGS ON STEEL SUBTRACTS BY A LOW-COST, FAST ELECTROCHEMICAL TECHNIQUE______177

THE JAPANESE INFLUENCE IN MODERN WESTERN ARCHITECTURE ─AN INTRODUCTION─______178

PURINERGIC SIGNALLING IN A MODEL OF OVERACTIVE URINARY BLADDER _ 179

CONDUCTION BLOCK IN ISOLATED RAT SCIATIC NERVE BY CHIRAL AMINOALKANOLIC DERIVATIVES OF XANTHONES______180

OSTEOCLASTIC RESORPTION OF CALCIUM PHOSPHATE BASED BONE SUBSTITUTES: IN VITRO STUDIES ______181

MONITORING PARTICULATE MATTER USING MAGNETIC PROPERTIES OF TREE LEAVES IN THE PORTO URBAN AREA AND CINFÃES AREA ______182

ADENOSINE REGULATES ITS OWN RELEASE FROM MYENTERIC NEURONS VIA A2A RECEPTORS COUPLED TO ADENYLATE CYCLASE / CAMP PATHWAY _____ 183

EDUCATION AND POVERTY: AN EMPOWERMENT APPROACH ______184

SYNTHESIS OF VESICULATED POLYESTER PARTICLES AND ITS INCORPORATION IN PAINTS ______185

CHARACTERIZATION OF UREA-FORMALDEHYDE RESINS ______186

CHARACTERISATION OF THE PLANCTOMYCETES PIRELLULA SP. OJF20 AND PIRELLULA SP. OJF27 ISOLATED FROM THE SURFACE OF MACROALGAE ____ 187

INVESTIGATION OF THE INTERACTIONS BETWEEN PHOSPHOLIPIDS AND GOLD NANOPARTICLES AT AIR/WATER INTERFACE ______188

DEVELOPMENT AND VALIDATION OF A NEW MULTIRESIDUE METHOD FOR THE DETERMINATION OF 17 POLYCHLORINATED DIBENZODIOXINS (DIOXINS) AND POLYCHLORINATED DIBENZOFURANS (FURANS) IN ENVIRONMENTAL MATRICES BY SPME-GC-MS ______189

ADENOSINE REGULATES DIFFERENTIATION OF HUMAN OSTEOBLAST CELLS IN CULTURE ______190

THE POMBAIS (PIGEON HOUSES) IN THE NORTHEAST TRANSMONTANO (NORTHEAST OF PORTUGAL) ______191

PRAVASTATIN QUANTIFICATION USING SQUARE-WAVE VOLTAMMETRY _____ 192

MICROBIAL COMMUNITY DISTRIBUTION AND DYNAMICS IN A SHALLOW RACEWAY-RECIRCULATING MARICULTURE SYSTEM ______193

INFORMATION SYSTEMS ABOUT MATERIALS FOR ARCHITECTURE______194

OSTEOGENIC POTENTIAL OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS. EFFECT OF MEDIUM COMPOSITION AND CELL PASSAGE ______195

POTENTIAL OF EQUISETUM ARVENSE HYDROMETHANOLIC EXTRACTS AS OSTEOBLASTIC GROWTH PROMOTERS ______196

COMPARISON OF THE IN VIVO BONE FORMING ACTIVITY OF OSTEOGENIC- INDUCED MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW______197

TAXONOMY FROM RURAL DIFFUSE TO URBAN INDUSTRIAL DIFFUSE______198

TETRACYCLINES MODULATION OF THE OSTEOGENIC DIFFERENTIATION – IN VITRO EVALUATION ______199

STUDY AND DESIGN OF PRECAST REINFORCED CONCRETE BOX-CULVERTS UNDER HIGH EMBANKMENTS ______200

PROPERTIES OF NANOSTRUCTURED MATERIALS ______201

INFLUENCE OF EGG YOLK AND AROMATIZING COMPOUNDS ON THE VOLATILE PROFILE AND ON SENSORY CHARACTERISTICS OF BAKERY CREAM ______202

EVALUATION OF GENETIC AND CHEMICAL DIVERSITY IN PORTUGUESE POPULATIONS OF PHASEOLUS VULGARIS L. ______203

THE METAMORPHOSIS OF ALEIXO TOWERS ______204

MODULATION OF CYTOCHROME P450 AND OXIDATIVE STRESS BY ROOIBOS (ASPALATHUS LINEARIS) INFUSION INTAKE ______205

SCREENING FOR NOVEL CDH1 INACTIVATING MECHANISMS IN FAMILIAL GASTRIC CANCER______206

THE INFLUENCE OF INTERPERSONAL SIMILARITY IN SOCIAL CATEGORIZATION ______207

GENETIC CHARACTERIZATION BY SSR OF SIX OLEA EUROPAEA REGIONAL VARIETIES FOR AUTHENTICITY EVALUATION OF OLIVE OILS ______208

PROGRAMA SOCIAL ______209

PLANTAS DO EDIFÍCIO DA FACULDADE DE ARQUITECTURA ______210

PROGRAMA GERAL

FEB. 20, WEDNESDAY FEB. 21, THURSDAY FEB. 22, FRIDAY 8:30 8:45 Registration 8:45 9:00 9:00 9:15 Welcome Session (A1+A2) 9:15 9:30 Parallel Oral 9:30 9:45 II.1 - FOOD & HEALTH (A1) Sessions II - (POS.II) 9:45 10:00 II.2 - HOUSE & LIVING (A2) 10:00 10:15 Interdisciplinary Oral Session 1 - IOS1 (A1+A2) II.3 - NANO & SUPRAMOLECULAR CHEMISTRY (A3) 10:15 10:30 Social Events 10:30 10:45 10:45 11:00 Coffee Break & Posters 11:00 11:15 Coffee Break 11:15 11:30

11:30 11:45

11:45 12:00 Parallel Oral

12:00 12:15 III.1 - LIFE & HEALTH II (A1) Sessions III - (POS.III) Interdisciplinary Oral Session 2 - IOS2 (A1+A2) 12:15 12:30 III.2 - ENVIRONMENT & TECHNICAL ASPECTS (A2) 12:30 12:45 III.3 - ARCHITECTURAL PERSPECTIVES (A3)

12:45 13:00 13:00 13:15

13:15 13:30 Lunch Break 13:30 Lunch Break 14:30

2

14:30 14:45 14:45 15:00 15:00 15:15 Workshop I (A1+A2) Workshop II (A1+A2) 15:15 15:30 15:30 15:45 15:45 16:00 16:00 16:15 Coffee Break 16:15 16:30 Coffee Break & Posters 16:30 16:45 16:45 17:00 17:00 17:15 Parallel Oral 17:15 17:30 Parallel Oral IV.1 - GREEN CHEMISTRY & ENVIRONMENT (A1) Sessions IV 17:30 17:45 I.1 - LIFE & HEALTH I (A1) Sessions I - (POS.I) IV.2 - LIFE & WORK & SOCIETY (A2) (POS.IV) Social Events 17:45 18:00 I.2 - TERRITORY & NETWORK (A2) IV.3 - LIFE & HEALTH III (A3) 18:00 18:15 I.3 - MODELLING & AUTOMATIC SYSTEMS (A3) 18:15 18:30 Closing Session of the Scientific Activities (A1+A2) 18:30 Porto de Honra Auditoriums: A1-A2-A3

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INTERDISCIPLINARY ORAL SESSIONS - IOS1 WEDNESDAY, FEB. 20

Manuela Aguiar & Georgina Correia da Silva What can a simulation study tell us about the relevance of utility and wealth indicators for decision 31 making concerning production and market behaviour Frederico Teixeira, Pavel B. Brazdil 62 Construction of a learning unit in a Moodle platform: studying mitosis using fruit fly as a model A. Barros, C. Melo, N. Regadas, J. Santos, S. Pereira 65 The Karimojong people from Uganda: a history assessed through patterns of mitochondrial DNA V. Pereira, L. Gusmão, A. Amorim, M. J. Prata 85 Angiogenesis modulation by catechin Duarte DD, Silva RP, Azevedo I, Soares R, Negrão R Novel functionalised textiles through nanostrucutured carbon and clay based materials 89 (NANOTEXTUP) I. Rocha, T. Galvão, A.G. Gonçalves, C. Pereira, F. Gonçalves, A. Ribeiro, A. Carneiro, M. F. R.

Pereira, C. Freire Alfonso X's General Estoria and the Portuguese Culture: an example of continuity between Middle 114 Ages and Modernity Mariana Soares da Cunha Leite

What can a simulation study tell us about the relevance of utility and wealth indicators for decision making concerning production and market behaviour

Frederico Teixeira1 and Pavel B. Brazdil 1,2

1 Faculty of Economics, University of Porto, Portugal 2 LIAAD – INESC Porto L.A., University of Porto, Portugal

In this article we discuss the importance of utility as a crucial parameter for economic decisions, in addiction to the more common parameter wealth. The work presented can be seen as belonging to the area of Computational Economics [1] and has been inspired by the work of Wellman [2]. Our study involves a simulated world with several agents which interact through a market. The agents have different quantities of different types of products, and its behaviour is driven by the goal to maximize the utility measure. This is calculated in an usual way [2] and takes into account the basket of goods detained by each agent. Wealth is calculated by simply multiplying basket quantities by current prices. An auctioneer determines the appropriate current price using several iterations, until equilibrium is established. In order to achieve their goal, agents exchange goods at the end of each period, and initiate a new period of production and consumption. Each agent produces only one type of product, but consumes certain quantities of all products. Production can increase/decrease by a fixed or proportional amount, while consumption is fixed. Our goal was to examine the trend of wealth and utility over time under different conditions, including, for instance, under-production of all goods. We have focused our attention on wealth, as normally this parameter is suggested for the control of production. We have verified that when fewer products are produced, the agents’ wealth remains more or less the same. The agents own less quantities, but those achieve higher prices. Surely, this is not a satisfactory situation for any of the agents and so we conclude that the parameter wealth by itself does not provide sufficient information to control production in a satisfactory manner. Furthermore, we carried out some experiments in which production was varied in a random manner, and observed that was possible to increase both wealth and utility. So, utility could help solving the problem. We conclude that utility and wealth can have different behaviours in identical situations, but that it is possible to increase both at a time, meaning that it is necessary to analyze them together and create rules that efficiently describes the agents’ global situation. Our future aim is to try different strategies that an agent could adopt regards production, using machine learning or other adaptive techniques to develop such rules. We intend to evaluate whether the information concerning the agents’ utility (i.e. not only wealth) is useful when controlling production, leading to better results overall. References [1] Tesfatsion, Leigh, K.Judd: Handbook of Computational Economics, Agent-based Computational Economics, Volume 2, North Holland, 2006. [2] Wellman, M., J.Hu: Conjectural Equilibrium in Multiagent Learning, Machine Learning, 33- 2/3, 1998.

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Construction of a learning unit in a Moodle platform: studying mitosis using fruit fly as a model

A. Barros 1, C. Melo2, N. Regadas2, J. Santos2, S. Pereira1,2

1Department of Botany, Faculty of Sciences, University of Porto, Portugal. 2Office for Science Communication, Institute for Molecular and Cell Biology (IBMC), University of Porto, Portugal.

The project “Construction of a learning unit in a Moodle plataform: studying mitosis using fruit fly as a model” funded by “Scientific Investigation in pre-graduation” of UP is a new strategy to engage students in a learning process. The aim of this project is the construction of a b-learning unit to be used in formal and non-formal education. It will be implemented in a biology educational site named “Estaleiro da Ciência”, coordinated by IBMC in collaboration with FCUP funded by Ciência Viva. The development of this learning unit, requires expertise in different domains being supported by an interdisciplinary team: a Pre-graduation student of Biology teaching, a master of Biology teaching and a graduate of Journalism and Communication Science of UP. The construction of this tool relies on Scientific, Didactics and Multimedia know-how. Biological material handling and experimental procedures were already defined and tested1in the Molecular Genetics Laboratory of IBMC. Didactics basis relies on the Teaching trough Research2 approach. In order to be implemented this tool will be organized in three stages: challenge, “hands-on” and results presentation. The first consists in a presentation of the challenge to the public in a way to motivate research and the engagement in the experimental activities. This will include various experimental reports with photos and video images. The practical work is a “Hands-on” approach using selected microscopy techniques. At last, results of experimental activities will be presented as a scientific article and/or a poster, with subsequent discussion between participants or other public on the web site. The multimedia component will conjugate different forms of information: video, animations, photos. Several functionalities are also being defined with the incorporation of this unit in the software Moodle, to promote the flexibility and the interactivity between users3. This unit, resulting of an articulation of the New Tecnologies of Information and Communication with Didactics can be a new idea intertwining Research Laboratories and Universities to improve Science Education and Scientific Outreach.

References: [1] Barros, I. (2007). Drosophila melanogaster como Modelo Biológico no Ensino da Mitose. Dissertação de Mestrado. Faculdade de Ciências da Universidade do Porto. Porto

[2] Cachapuz, F. et al (2000). Perspectivas de Ensino, Textos de Apoio n.º1, Formação de Professores – Ciências. Centro de Estudos de Educação em Ciência. Porto.

[3] Melo, C. (2006). Promoção do Ensino Experimental das Ciências: Construção e Integração de Material Didáctico num Software Educativo, na temática Reprodução sexuada. Dissertação de Mestrado. Faculdade de Ciências da Universidade do Porto. Porto.

6

The Karimojong people from Uganda: a history assessed through patterns of mitochondrial DNA

V. Pereira 1, 2, L. Gusmão 2, A. Amorim 1, 2 and M. J. Prata 1, 2

1 Department of Zoology and Anthropology, Faculty of Sciences, University of Porto, Portugal. 2 Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP).

Patterns of genetic diversity in populations are influenced by demographic factors such as fluctuation in population size and structure, admixture and migration. Many can be reconstructed from mtDNA and Y chromosome analysis. One of the most complex demographic events that shaped Africa’s history was the Bantu dispersal which started around 5000 y.a. and was responsible for the introduction of agriculture in sub-Saharan regions that were previously inhabited by tribes of hunter-gatherers. The Eastern branch of the expansion is said to have reached Uganda near 1000 BC [1]. The impact of this people movement on sub-Saharan populations was so strong that practically obscured other less pronounced events. This was the case of the Nilotic migration that introduced cattle- herding techniques in many regions of Eastern Africa. The Karimojong are one of the groups from this movement presently established in Northern Uganda. In this study sequence data from mtDNA hypervariable segments were obtained for 55 Karimojong unrelated individuals. Diversity at HVS I, II and III from mtDNA was determined through direct sequencing methods. The samples were classified into haplogroups according to recommended nomenclature [2]. Further statistical analyses and comparisons with other african populations were performed. The results showed that Karimojong have a high diversity level as typically found in East African populations. 91% of the samples belonged to macrohaplogroup L which is specific of sub-Saharan regions. Some of the haplogroups detected occur commonly in Bantu groups; others are shared by a number of Nilotic groups that supposedly have a common origin. The presence of rare haplogroups in frequencies higher than those usually found in East Africa may be a sign of the genetic profile ancestral to the Nilotic populations.

References: [1] Ehret, C. (2001), Bantu Expansions: Re-Envisioning a Central Problem of Early African History, The International Journal of African Historical Studies 34:5-41. [2] Bandelt, H-J. et al. (2006), Human Mitochondrial DNA and the evolution of Homo sapiens, Springer, Heidelberg, Germany.

7

Angiogenesis modulation by catechin

Duarte DD, Silva RP, Azevedo I, Soares R and Negrão R

Department of Biochemistry (U38/FCT), Faculty of Medicine, University of Porto, Portugal.

Angiogenesis, the formation of new blood vessels from pre-existing ones, is a process that occurs in many physiological conditions, like wound healing and pregnancy, but also in pathological situations such as cancer and cardiovascular disease. Deficient angiogenesis results in deficient blood flow as in coronary heart disease and acute myocardial infarction. Recent Western diet is believed to contribute to an increased lifetime risk of cancer and cardiovascular disorders. On the other hand, diets high in plant-derived foods offer a protective effect. Recent studies indicate that the development of these pathologies is inversely associated with the consumption of natural polyphenolic compounds, which are known to affect angiogenesis. Identification and characterization of dietary phytochemicals able to block, slow or reverse angiogenesis may, thus, constitute an important strategy for prevention of cancer and cardiovascular disease. Catechin is a bioactive phytochemical abundant in some beverages like tea and wine and also in fruits and vegetables. The purpose of this study was to investigate the effect of catechin in angiogenesis, namely evaluating its effects on human umbilical vein endothelial cells (HUVEC) and human aortic smooth muscle cells (HASMC) viability, apoptosis, proliferation, migration, invasion and also capillary-like structures formation. Treatment of cells with 10-100 µM catechin resulted in a significant increase in both HUVEC (165.34±31.12%) and HASMC (165.58±5.04%) viability assessed by MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and confirmed by measuring lactate dehydrogenase activity released to the extracellular medium. Furthermore, the number of apoptotic cells determined by TUNEL (Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling) assay diminished slightly in HUVEC (to 46.55±12.88%), and drastically in HASMC (to 7.35±4.85%). Proliferation evaluation by measuring Brdu cellular incorporation as well as determination of cell invasive capacity using Transwell BD-Matrigel basement membrane matrix insert are still under analysis. Catechin treatment led to a slight reduction in HASMC cell migration to injured areas (injury assay) but it seemed to increase HUVEC migration in the same conditions. Incubation of HUVEC on growth factor reduced- Matrigel-coated plates for 24 h with 10 µM catechin led to the formation of highly ramified cord-like structures (174.07% ±14.93%). The results obtained so far indicate that catechin decreased cell apoptosis and exerted stimulatory effects on endothelial and vascular smooth muscle cell’s viability and on HUVEC migration and capillary-like structures formation, processes that are required for the development of physiological and pathological angiogenesis.

Supported by ERAB (EA0641) and FCT (POCTI/SAU-BMA/55556/2004).

8

Novel functionalised textiles through nanostrucutured carbon and clay based materials (NANOTEXTUP)

I. Rocha 1, T. Galvão1,A.G. Gonçalves,2 C. Pereira1, F. Gonçalves,2 A. Ribeiro,3 A. Carneiro,3 M. F. R. Pereira2 C. Freire1

1 REQUIMTE, Departamento de Química, Faculdade de Ciências, Universidade do Porto, 4169- 007 Porto. 2 Laboratório de Catálise e Materiais, Departamento de Engenharia Química, Faculdade de Engenharia, Universidade do Porto, 4200-465 Porto. 3 Centro de Nanotecnologia e Materiais Técnicos, Funcionais, e Inteligentes, 4760-034 Vila Nova de Famalicão

Textiles are probably the most popular of materials since they decorate and protect our bodies while bringing comfort into our lives. Currently, the consumers are demanding for textile products with higher performances. A significant product differentiation can be achieved by incorporation of high performance properties, which are beyond the pure aesthetic and decorative functions. They include not only a large range of properties, but also multiple functions, leading to the so-called (multi)functional textiles. The functionalisation of textiles have been made using nanotechnology as these endeavours are mainly focused on using nanosized substances and generating nanostructures during manufacturing and finishing processes. In this project we will attempt to produce cotton- based functional textiles with improved stability against mechanical and thermal (fire) destruction by functionalisation with clay nanoparticles, carbon nanotubes and nanofibers.

+

Nanomaterials Textile substrate Functional textiles

The work has started with the functionalisation of nanomaterials to be grafted to cotton fibers. CNTs were functionalised by oxidation in the liquid phase with HNO3 (introduction of acid groups), oxidation in the gas phase with O2 (introduction of neutral and basic groups), and heat treatments at different temperatures to selectively remove some of the functional groups. Some of these carbon materials will be then functionalised with organosilanes which will react with some of the functional groups that will be present in the cotton substrates. These samples were characterised by TPD. The clays, montmorillonite (natural clay) and laponite (synthetic clay), were functionalized with two organosilanes with reactive bromide and chloride groups and these materials were made to react with cotton substrates. In another approach, silica nanoparticles functionalised with amine groups were grafted to cotton substrates by a one-step methodology. All the materials were characterized by FTIR and SEM. Acknowledgments: This work is funded by Proj. IPG63 de Investigação Científica na Pré- Graduação 2007 (Universidade do Porto e Caixa Geral de Depósitos).

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Alfonso X’s General Estoria and the Portuguese Culture: an example of continuity between Middle Ages and Modernity

Mariana Soares da Cunha Leite 1

1 Master of Arts Student in Romanic Literatures, Department of Portuguese and Romanic Studies of the Faculdade de Letras da Universidade do Porto, Portugal. Researcher of the Medieval Seminar of Literature, Thought and Society, Instituto de Filosofia of the Faculdade de Letras da Universidade do Porto, Portugal

This paper aims to present a synthesis of the research work I have been conducting for the past year in the field of medieval literature and medieval Iberian culture. It will culminate in a Master’s degree thesis, due late this year. Historiography is, beyond doubt, a privileged means of understanding cultures, world visions and Men’s attitudes towards their past. Within Iberian vernacular historiographic literature, Alfonso X’s 1252-88 General Estoria (of which only the two first chapters1 and fragments of the third2 were edited – a circumstance that heavily hinders the study of the text) occupies a particularly conspicuous place. His ambitious project of writing a history of the world from its remote creation until the birth of Jesus capable of encompassing both the Greek-Roman and the Judaic-Christian historical traditions, had an impact on Portuguese culture whose profound consequences have not yet been fully studied. There existed at least two translations: one into Portuguese3, fragmentary, and the other into Galician4, comprising only the first two parts. Due to the superficiality of the studies produced so far regarding these narratives, we still ignore which parts of them were more decisive in influencing Portuguese culture – not only in what concerns historiography, but literate culture in general. In view of this, and bearing in mind the difficulties posed by the fragmentary nature of the General Estoria, I will try to analyse the diffusion in Portugal of the Alfonsi text, taking as example an unpublished manuscript kept at the Biblioteca Pública de Évora. The excerpt I will be working upon is part of a larger edition of the manuscript, which will be included in my thesis. In order to make my point more clearly, I will provide a document with the transcription.

References: [1] Solalinde, António G. [ed.] (1930), Alfonso X, General Estoria. Madrid, (s/n). See also: Sánchez-Prieto Borja, Pedro [ed.] (2001), Alfonso X, General estoria.Madrid, Fundación José Antonio de Castro. [2] Sánchez-Prieto Borja, Pedro; Horcajada Diezma, Bautista [ed.] (1994), Alfonso X, General estoria. Tercera parte. Madrid, Gredos. See also: Fernández López, María del Carmen [ed.] (1998), Edición crítica del “Libro de Isaías” de la Tercera Parte de la “General Estoria”. Alcalá de Henares, Servicio de Publicaciones de la Universidad de Alcalá de Henares [3] Costa, Avelino Jesus da (1949), Fragmentos preciosos de códices medievais, Braga, Bracara Augusta. [4] Martinez-Lopez, Ramón [ed.] (1963), Alfonso X, General Estoria : versión gallega del siglo XIV : Ms. .I.I. del Escorial. Oviedo, Facultad de Filosofia y Letras, Universidad de Oviedo.

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INTERDISCIPLINARY ORAL SESSIONS – IOS2 WEDNESDAY, FEB. 20

Lígia Afonso & Luís Miguel Madeira 228 Alternative User Interfaces for Game Control Sarah Imboden 119 Traditional Timber Construction Buildings: The example of the Portuguese Central Littoral Palheiros D. Moutinho, J. Teixeira Treatment of Myasthenia gravis with cholinesterase inhibitors: on the role of nicotinic autoreceptors 164 containing a3b2 subunits Ana Sá-e-Sousa, Tiago Morais, Diogo Trigo, Maria Alexandrina Timóteo, Bernardo Matos, Laura

Oliveira, P. Correia-de-Sá 188 Music and Phonological Awareness L. Saraiva, J. Machado-Vaz, S. Rodrigues, S. Monteiro 190 Automatic Emotion Assessment through Biometric Information J. Teixeira, V. Vinhas Development of molecular biology techniques for the detection of genetically modified organisms in 193 maize food products S.C. Sousa, I. Mafra, C.S. Ferreira da Silva, J.A. Amaral, M.B.P.P. Oliveira Permeability Characteristics of High Performance Concrete (HPC) Containing Varying 221 Concentrations of Hycrete Tested and Compared by Method of Rapid Chloride Permeability Test and the Non-Steady-State Migration Test (nordtest method NT Build 492) Michael Boxer, Hani Nassif 224 Mario Pedrosa e sua atuação crítica na Presidência da ABCA Gabriela Borges Abraços

Alternative User Interfaces for Game Control

Sarah Imboden

University of Texas at Austin

12

Traditional Timber Construction Buildings: The example of the Portuguese Central Littoral Palheiros

D. Moutinho 1 and J. Teixeira 2

1 Department of Architecture, Faculty of Architecture, University of Porto, Portugal. 2 Department of Architecture, Faculty of Architecture, University of Porto, Portugal.

The palheiros are traditional timber construction buildings on the Portuguese Central Littoral. They were originally conceived to store the tools related with fishing activity and to give shelter for the fishermen during the fishing season. Located near the sea, they started to be also used as holiday homes. The owners with more income built their own dwelling, mostly with a bourgeois look, more comfortable and vigorous than the fisherman’s one. Most of the existing information about these buildings is obsolete, based on the reality of the middle 20th century, mostly descripted on ethnographic publications [1]. Concerning this fact, we found necessary to update and amplify the knowledge about this subject, on the architectural and constructive point of view, regarding the found examples. These buildings are the result of a continuous evolving process which ended on a variety of forms and technical solutions, during the last few centuries. However, the majority looks very wrecked and damaged by the application of industrial materials and techniques, during the last century, breaking up with their ancient timber technique. Since we don’t know for how long these palheiros will remain, our concern is to make the inventory of the exiting ones and to learn about their original construction system. We found unique and ingenious solutions for everlasting architectural issues. Yet, among the variety of cases, which are the result of each owner’s construction skills, we have detected a huge number of resemblances, revealing a common starting point, a raison d'être, a consistent technique, a tradition. With this research, we give our contribute appealing to the documentation and analysis, preservation and rehabilitation of these unique examples of Portuguese timber vernacular architecture.

[1] Oliveira, E.V. and Galhano, F. (1964), Palheiros do Litoral Central Português, Instituto de Alta Cultura, Lisboa.

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Treatment of Myasthenia gravis with cholinesterase inhibitors: on the role of nicotinic autoreceptors containing α3β2 subunits

Ana Sá-e-Sousa1, Tiago Morais1, Diogo Trigo1, Maria Alexandrina Timóteo1, Bernardo Matos1, Laura Oliveira1 & P. Correia-de-Sá1

1Laboratório de Farmacologia e Neurobiologia, Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas de Abel Salazar – Universidade do Porto (ICBAS-UP), Portugal.

Failure of neuromuscular transmission is rare due to excess of acetylcholine (ACh) release to levels higher than those required to trigger an action potential at the muscular fiber (i.e. high safety factor). Transmission safety factor may be limited in several pathological conditions, namely in Myasthenia gravis. So far, most attempts to improve muscle weakness that characterize this disorder have been to prevent ACh breakdown by blocking cholinesterase activity. Recently, we demonstrated that the neuromuscular junction is equipped with α3β2-containing nicotinic autoreceptors (nAChRs) mediating facilitation of ACh release, which might also contribute to the therapeutic action of cholinesterase inhibitors [1]. This prompted us to investigate the effect of neostigmine (Neo, a commonly used cholinesterase inhibitor) and dihydro-β-erythroidine (DH-β-E, a relatively weak antagonist at muscle-type α1 nicotinic receptor as compared to the α3β2 subtype) on nerve- evoked contractions and [3H]-ACh release from motor nerve terminals of rats with Toxin- Induced Myasthenia gravis (TIMG). Wistar rats (70-100 g) were injected once every 48h with saline (controls) or α- bungarotoxin (α-BTX, an irreversible muscle-type α1 nicotinic receptor antagonist) (TIMG-model) for a period up to 6 weeks [2]. Dosage of α-BTX was adjusted by monitoring myasthenic symptoms. [3H]ACh release was evoked by phrenic nerve stimulation with 50 Hz-bursts (5 bursts of 150 pulses, with a 20s interburst interval). Fatigue tests were carried out using high frequency (50 Hz) intermittent (17 pulses per sec, during 3 minutes) nerve stimulation. Chronic injections of α-BTX decreased the amplitude of nerve-evoked diaphragm contractions during the fatigue tests. Neo (500 nM) transiently (30s) increased diaphragm contractile responses in both control and TIMG animals. Following the initial 30-sec stimulation period, Neo (500 nM) significantly (P<0.05) enhanced fatigue of muscle contractions in controls, but it was virtually unchanged in TIMG animals. Neo (500 nM) decreased nerve-evoked (50 Hz-bursts) [3H]-ACh release by 58±8% (n=6) in control animals, but the magnitude of this effect was reduced to 13±7% (n=7) in TIMG animals. Blockade of α3β2-containing neuronal nAChRs with DH-β-E (3 μM) enhanced fatigue in control animals, but it was virtually devoid of effect in TIMG animals. Inhibition of [3H]- ACh release by DH-β-E (3 μM) was more evident in TIMG animals (-25±6%, n=9) than in controls (-9±4%, n=5). Pre-treatment with DH-β-E (3 μM) abolished Neo (500 nM) improvement of muscle contraction in both control and TIMG animals. Data suggest that activation of facilitatory α3β2-containing nicotinic autoreceptors minimize neuromuscular deficits in rats with toxin-induced Myasthenia gravis by increasing the safety margin of synaptic transmission. [1] Faria et al. (2003), Synapse 49 ,77-88 ; [2] Plomp et al.(1992), J. Physiol. 458, 487-499. This work was supported by FCT (FEDER funding, POCTI/CVT/43368/2001).

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Music and Phonological Awareness

L. Saraiva 1, J. Machado-Vaz1, S. Rodrigues1, S. Monteiro1

1 Department of Psychology, Faculty of Psychology and Sciences of Education, University of Porto, Portugal.

This work about Psychology of Language and Cognition refers to the development of an interactive and multimedia instrument that analyses the acquired skills in the field of phonological awareness. A cd-rom was prepared for pre-school children with the aim to help them learn how to read and write. Nevertheless it is destined also to children with phonological awareness deficits, whether or not as part of other kinds of problems (autism, dislexia). The researched bibliography leads us to the conclusion that phonological awareness is one of the main determinants of the reading and writing skills in children; the ability to sort similarities and differences between sounds is closely related to musical skills so, understanding the benefits of musical learning in the development of the skill to distinguish phonemes, particularly through pitch differences, we created a little story with songs made according to criteria of phonological likeness. The different sound categories are presented in a musical media, and are subjected to tasks of sound categorization, recognition and segmentation. The songs and lyrics that were created to each sound class highlight certain sounds that need to be worked on through melody, rhythm and musical structure. Due to the interactivity of the cd-rom, the child is able to participate directly in the exercises, which consist in the correct association between a phoneme and its grapheme. In conclusion, this cd-rom develops the skill to understand that the written word is a way of representing the sound properties of the spoken word, so that reading becomes more amusing and interesting.

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Automatic Emotion Assessment through Biometric Information

J. Teixeira1, V. Vinhas1

1 Department of Electrical and Computer Engineering, Faculty of Engineering, University of Porto, Portugal. LIACC – Artificial Intelligence and Computer Science Laboratory Porto, Portugal {teixeira.jorge, vasco.vinhas}@fe.up.pt

While affective computing and the entertainment industry still maintain a substantial gap between themselves, biosignals are subject of digital acquisition through low budget technologic solutions at neglectable invasive levels preventing users from focusing their awareness in the equipment. Having this macrocontextualization in mind, the presented project has the main intention of using emotion assessment through biosignals to promote both subconscious interaction and individual specific appropriate content delivery. The proposed system constitutes a solid technologic framework that intends to enable biological information acquisition in a controlled environment having as initial hypothesis the existence of human physical expression of emotional states that can be objectively measured by relatively inexpensive equipment. The integration of electroencephalography, galvanic skin response and oximeter in a multichannel framework constitutes an effort in the path to identify emotional states via biosignals expression [1]. The flexibility of this system is one of the most important characteristics so that it enables the integration of a diversity of biometric equipment in order to improve the quality of the emotions’ assessment. In order to guarantee the control of the induced emotional states and optimize the biometric equipments available, the induction through visual stimuli [2] was the most suitable method for this project, so that gender specific sessions stratified accordingly to its valence value [3] were defined based on the International Affective Picture System [4]. The experimental conditions were performed in a controlled environment under very restricted exclusion criteria [5]. An automatic emotions’ assessment toll is able to gather the pre-processed and analyzed biometric information and identify the emotional states triggered by the visual stimuli. The attained success levels concerning relating emotions to biosignals are extremely encouraging not only to the continuation of this research topic but also to the application of these results in domains such as multimedia entertainment, advertising and medical treatments. References: [1] K. Ishino, M. H. (2003). A feeling estimation system using a simple electroencephalograph. In Proceedings of 2003 IEEE International Conference on Systems, Man, and Cybernetics, pages 4204–4209. [2] G. Chanel, J. Kronegg, D. G. (2005). Emotion assessment: Arousal evaluation using EEG’s and peripheral physiological signals. In Technical Report. [3] L. Aftanas, N. R. (2006). Neurophysiological correlates of induced discrete emotions in humans: An individually oriented analysis. In Neuroscience and Behavioral Physiology, vol 36. [4] P.J. Lang, M. Bradley, B. C. (2005). International affective picture system (IAPS): Affective ratings of pictures and instruction manual. In Technical Report. University of Florida [5] R. Paul, C. C. (2005). Age-dependent change in executive function and gamma 40 hz phase synchrony. In Journal of Integrative Neuroscience, volume 4, pages 63–76.

16

Development of molecular biology techniques for the detection of genetically modified organisms in maize food products

S.C. Sousa 1, I. Mafra1, C.S. Ferreira da Silva1, J.A. Amaral1, M.B.P.P. Oliveira1

1 REQUIMTE-Laboratory of Bromatology, Faculty of Pharmacy, University of Porto, Portugal.

In the last years, the increase in the cultivated area of genetically modified (GM) maize has become a reality. GA21, MON810 and MON863 maize crops are some of the authorized maize events for food and feed under the European Union (EU) regulations. These crops of transgenic maize bring profit towards the conventional ones, as they confer resistance to some plagues and/or herbicides [1]. Concerning the raise of production and consumption of foodstuffs derived from genetically modified organisms (GMO), the EU has established new demand levels, including the labeling requirements when the product has GMO in a proportion higher than 0.9% (Regulation (EC) N.º 1829/2003). The need to monitor and verify the presence of biotechnology-derived material in food products demands analytical methods able to detect, to identify and to quantify either the introduced DNA or the expressed protein(s). The DNA based methods, namely the Polymerase Chain Reaction (PCR) showed to be tools of great specifity and sensitivity in the analytical control concerning the presence of GMO [1]. The goal of this work was to apply and develop PCR techniques for the detection of GM maize in raw and processed foodstuffs. The first step was the DNA extraction of the samples by two methods: CTAB and/or Wizard [2]. Yield and purity of DNA extracts were assessed by spectrophotometry, while amplifiability was evaluated by PCR targeting the invertase gene. The screening of GMO was performed by the detection of 35S promoter from the cauliflower virus. The specific detection of GMO events, such as GA21 maize, MON810 maize and MON863 maize was carried out by PCR techniques. The results of DNA extraction showed that the CTAB method gave higher purity and DNA amplifiability in some of the samples, meaning that those extracts were more suitable for PCR amplification. However, despite the lower purity of extracts, the Wizard method gave generally higher DNA yields. The results of 35S screening sequence by PCR did not show any apparent positive sample. However, the PCR for 35S showed low sensitivity, although attempts where done to improve it. In the detection of specific GMO events, there was one positive result for the GA21 maize, three positive samples for the MON810 maize and no positive result for MON863.

References: [1] Mafra I., Oliveira M.B.P.P. (2005), Organismos Geneticamente Modificados e Alimentos derivados: II. Métodos de Detecção e Quantificação, Alimentação Humana, 11(2), 71-86. [2] Mafra, I., Silva, S.A., Moreira, E.J.M.O., Ferreira da Silva, C.S., Oliveira, M.B.P.P. Comparative study of DNA extraction methods for soybean derived food products, Food Control (submitted).

17

Permeability Characteristics of High Performance Concrete (HPC) Containing Varying Concentrations of Hycrete Tested and Compared by Method of Rapid Chloride Permeability Test and the Non-Steady-State Migration Test (nordtest method NT Build 492)

Michael Boxer, Hani Nassif

Civil Engineering Department, Rutgers The State University of New Jersey

The durability of concrete structures has become a huge concern to the public with media responses to multiple bridge collapses in recent times. Permeability of concrete causes chlorides to penetrate the reinforcing steel causing the concrete to become weak and ultimately failing. Through the use of the rapid chloride permeability test (ASTM C 1202- 05) and the nordtest (NT BUILD 492) the permeability of concrete can be determined. In recent times, the reduction of the permeability of concrete has been done by adding pozzolans and silica fume which are expensive additives to concrete. However, this study is on the effectiveness of a chemical admixture, Hycrete, because it is an admixture which is supposed to significantly reduce the permeability of concrete without having to use expensive high performance concrete. Although this study found that the addition of Hycrete did lower the permeability of concrete significantly in the high performance concrete mix we tested, this chemical admixture may be more effective in a class A concrete. Also this study used 3/8” coarse aggregate instead of the industry standard ¾’ for bridge decks. However, with the use of this chemical admixture, the durability of concrete structures can be increased and in the long run save money on construction jobs in harsh environmental conditions.

18

Mario Pedrosa e a sua atuação crítica na Presidência da ABCA

Gabriela Borges Abraços, Lisbeth Rebollo

Escola de Comunicação e Artes Universidade de S. Paulo, Brasil

19

PARALLEL ORAL SESSIONS I – POS.I.1 WEDNESDAY, FEB. 20

I.1 - LIFE & HEALTH I Maria José Oliveira & Adrián M.T. Silva Yeast as a new approach to study the regulation of p53-dependet apoptosis by protein kinase C 41 delta E. Cavaleiro, I. Coutinho, G. Pereira, J. Gonçalves, M. Côrte-Real, L. Saraiva 105 Antitumor and antifungal activities of xanthonic derivatives, potential inhibitors of steroid sulfatase E. Costa, E. Sousa, N. Nazareth, M. S. J. Nascimento, L. Vale-Silva, E. Pinto, M. Pinto 116 E-cadherin silencing: More than inactivating mutations Hugo Pinheiro, Joana Carvalho, Patrícia Inácio, Sónia Sousa, Elia Stupka, David Huntsman, Raquel Seruca, Carla Oliveira 128 Synthesis and Determination of Physicochemical Parameters of Xanthones C. Azevedo, C. M. M. Afonso, M. M. M. Pinto, J. L. F. C. Lima, S. Reis Dissemination of blaTEM-52 among Escherichia coli and Klebsiella pneumoniae strains is caused 159 by spread of plasmids in Portugal A. Pedrosa, A. Novais, E. Machado, R. Cantón, T. M. Coque, L. Peixe Synergism between adenosine A2A and tachykinin NK1 receptors facilitate [3H]-ACh release from 174 myenteric neurons I. Silva, C. Vieira, M. Duarte-Araújo & P. Correia-de-Sá

Yeast as a new approach to study the regulation of p53- dependent apoptosis by protein kinase C delta

E. Cavaleiro1, I. Coutinho1, G. Pereira1, J. Gonçalves1, M. Côrte-Real2, L. Saraiva1

1REQUIMTE, Laboratório de Microbiologia e Farmacologia, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha, 4050-030 Porto 2Centro de Biologia, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal

One promising approach to circumvent the apoptosis-resistance of tumour cells is through modulation of p53 tumour suppressor protein or of key components of its signalling pathway. In fact, p53 is considered the most relevant protein in apoptosis and, therefore, in carcinogenesis [1]. Nevertheless, the mechanisms underlying p53- mediated apoptosis are not well clarified [2]. Recent studies revealed that posttranslational modification, by phosphorylation, of p53 constitutes an important mechanism of p53 regulation, representing a novel targeted approach in cancer therapy [3]. These studies demonstrated that isoform delta of protein kinase C (PKC-delta) phosphorylates and, consequently, activates p53. Thus, agents that interfere with PKC-delta activity may modulate p53 responses and, therefore, can be used to increase the efficacy of chemotherapeutic drugs targeting p53 [4]. In order to confirm a possible regulation of p53 by PKC-delta, mammalian wild-type p53 and PKC-delta were co-expressed in the yeast Saccharomyces cerevisiae. As reported [5], we also confirmed that expression of p53 in yeast induces a marked growth inhibition, without causing cell death. We also detected that PKC-delta significantly increases p53- induced yeast growth inhibition. The effect of PKC-delta on p53 activity was also analysed in yeast treated with the apoptotic inducer H2O2. The results obtained showed that PKC- delta significantly increases p53-stimulation of H2O2-induced cell death, an effect accompanied by preservation of cell membrane integrity and by an increase on DNA fragmentation, reactive oxygen species production and caspase activation. We also confirmed that activators of PKC-delta, such as phorbol-12-myristate-13-acetate (PMA), can increase p53-induced apoptosis and that activation of p53-dependent apoptosis by the chemotherapeutic agent etoposide involves a PKC-delta pathway. References: [1] Wang, W. Rastinejad, F. and El-Deiry, W.S. (2003), Restoring p53-dependent tumor suppression. Cancer Biol Ther. 2(1), S55-63. [2] Brown, J.M. and Wouters, B.G. (1999), Apoptosis, p53, and tumor cell sensitivity to anticancer agents. Cancer Res. 59(7), 1391-9. [3] Yoshida, K. Liu, H. and Miki, Y. (2006), Protein kinase C delta regulates Ser46 phosphorylation of p53 tumor suppressor in the apoptotic response to DNA damage. J. Biol. Chem. 281(9), 5734-40. [4] Abbas, T. White, D. Hui, L. Yoshida, K. Foster, D.A. and Bargonetti, J. (2004), Inhibition of human p53 basal transcription by down-regulation of protein kinase Cdelta. J. Biol. Chem. 279(11), 9970-7. [5]Nigro, J.M. Sikorski, R. Reed, S.I. and Vogelstein, B. (1992), Human p53 and CDC2Hs genes combine to inhibit the proliferation of Saccharomyces cerevisiae. Mol Cell Biol. 12(3), 1357-65. We thank to REQUIMTE/CEQUP and to Universidade do Porto for financial support.

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Antitumor and antifungal activities of xanthonic derivatives, potential inhibitors of steroid sulfatase

E. Costa 1,3, E. Sousa1,3, N. Nazareth2,3, M. S. J. Nascimento2,3, L. Vale-Silva2,3, E. Pinto2,3 and M. Pinto1,3

1 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 Department of Microbiology, Faculty of Pharmacy, University of Porto, Portugal. 3 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal.

The enzyme steroid sulfatase (STS) has received increasing attention as a drug target, due to its role in the pathogenesis of several diseases, in particular estrogen-dependent tumors, with STS inhibitors entering in preclinical phases [1]. This work includes the synthesis of rigid analogues of the potent inhibitor of STS, BENZOMATE (1): xanthone-3,6-O,O-bis(sulfate) (7), 1-hydroxyxanthone-2-O-sulfate (8) and xanthone-3,4-O,O-bis(sulfate) (9). Other two precursors/analogues were obtained, the 3,6-dihydroxyxanthone (2) and the 2,2’,4,4’-tetracethylbenzophenone (6). These five derivatives (2, 6-9) as well as the raw material 5 were investigated for their antifungal activity and capacity to inhibit the in vitro growth of human tumor cell lines.

O O O OH O OH H NO SO 2 2 OSO2NH2 HO O OH 1 O O OH 2 OH 3 4

O O O - O O OSOH 3 - OSO3 - OSO - HO OH AcO OAc -O3SO O OSO3 O 3 OH OH OAc OAc O 7 OSO - 5 6 8 9 3

The synthetic pathway from 2,2’,4,4’-tetrahydroxybenzophenone (5), through dry heating in furnace (180ºC) afforded 3,6-dihydroxyxanthone (2) in a very good yield (85%). Dihydroxyxanthones 2-4 were treated with sulfur trioxide-pyridine in dimethylacethamide to furnish the xanthonic sulfate derivatives, 7, 8 and 9. The effect of compounds 2-9 on the growth of MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer) and SF-268 (CNS carcinoma) cell lines was evaluated. 3,6- Dihydroxyxanthone (2), 2,2’,4,4’-tetrahydroxybenzophenone (5) and 2,2’,4,4’- tetracethylbenzophenone (6) were shown to be active on all tumor cell lines (GI50‹150 μM). Regarding antifungal activity investigation, compounds 2, 5 and 6 showed a slight inhibitory effect against Microsporum gypseum and Trichophyton mentagrophytes at 125 μg/ml. References: [1] Nussbaumer P. and Billich A. (2004), Steroid Sulfatase Inhibitors, Medicinal Research Reviews, 24, 529-576. Acknowledgments: FCT (I&D, nº226/94), FEDER, POCI for financial support.

22

E-cadherin silencing: More than inactivating mutations

Hugo Pinheiro1, Joana Carvalho1, Patrícia Inácio1, Sónia Sousa1, Elia Stupka2, David Huntsman3, Raquel Seruca1,4, Carla Oliveira1,4

1IPATIMUP, Porto, Portugal; 2CBM, Trieste, Italy; 3British Columbia Cancer Agency, Vancouver, Canada; 4Faculty of Medicine, University of Porto, Portugal

INTRODUCTION: E-cadherin is a cell-to-cell adhesion transmembrane glycoprotein and a tumour suppressor. E-Cadherin (CDH1) inactivating germline and somatic mutations occur in hereditary and sporadic forms of diffuse gastric and lobular breast cancers. Independently of CDH1 mutations (somatic and germline), almost all diffuse-growing cancers display mislocalized or absent E-cadherin immunoexpression. This similarity suggests that still unidentified mechanisms may underlie E-cadherin expression impairment in mutation negative cases. In order to clarify this hypothesis we aimed at: 1) Developing a CDH1 allele-specific expression (ASE) method to determine allele specific expression imbalance; 2) Determining the frequency of CDH1 germline promoter methylation; 3) Addressing the existence of new transcribed portions in CDH1 and characterize their expression; and 4) Testing the putative in vitro functional role of the transcripts found. MATERIAL AND METHODS: Blood RNA was analysed for CDH1 ASE using a polymorphic site from: 17 normal controls, 21 CDH1 negative gastric cancer (GC) probands. Blood DNA from 68 GC probands was analysed for CDH1 germline promoter methylation. The full CDH1 sequence was submitted to GENSCAN and putative transcribed sequences within introns were predicted and its expression confirmed by RT- PCR. RESULTS: CDH1 ASE analysis showed: 1) in controls, equivalent expression of both alleles (1.48 ± 0.22), and; 2) in CDH1 negative probands, monoallelic CDH1 expression or allelic imbalance in 73% of the cases. Germline promoter hypermethylation was found in 1/68 (1.47 %) probands. Moreover, we found that CDH1 intron 2 contains expressed putative regulatory regions. DISCUSSION AND CONCLUSIONS: CDH1 allelic imbalance is highly frequent among CDH1 negative probands arguing towards the existence of unknown molecular mechanisms leading to monoallelic downregulation in CDH1 expression. The epimutation described seems to constitute a rare phenomenon and should not be considered to explain a significant proportion of GC cases without molecular diagnostic. The two expressed putative regulatory regions identified within CDH1 gene, outside of the currently known coding regions, may constitute target sequences for epi/genetic control leading to CDH1 expression downregulation.

23

Synthesis and Determination of Physicochemical Parameters of Xanthones

C. Azevedo 1,2, C. M. M. Afonso2, M. M. M. Pinto2, J. L. F. C. Lima1 and S. Reis1

1 REQUIMTE – Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto 2 CEQOFFUP – Serviço de Química Orgânica, Faculdade de Farmácia, Universidade do Porto. Rua Aníbal Cunha 164, 4050-047 Porto, Portugal

Natural and synthetic xanthones have revealed a vast diversity of biological activities and the xanthone nucleus has shown to be a good platform for the development of new pharmacological active compounds [1]. To transform a biological active compound in a successful drug it requires, among several other procedures, the synthesis of new molecules and the understanding of both pharmacodynamic and pharmacokinetic behaviours [2]. The objective of this work was to synthesize two series of xanthones and to evaluate their physicochemical parameters in order to establish models of pharmaceutical behaviour for xanthone compounds. It was synthesized, by classic methodologies and with the aid of microwaves, series of mono-hydroxylated and mono-methoxylated xanthones. The parameters studied were the partition coefficients and the acidity constants. So, it was possible to study the influence of two types of substituents in the xanthone nucleous: the hydroxyl (polar) and methoxyl (less polar) groups. The partition coefficient of the xanthones was studied in micelles of hexadecylphosphocoline (HDPC), and determined by derivative spectroscopy [3]. In the mono-hydroxylated series, the more lipophilic compound was 2-hydroxyxanthone and less lipophilic compounds were 3-hydroxyxanthone and 4-hydroxyxanthone. In the monomethoxylated series, the more lipophilic compounds were 2-methoxyxanthone and 3- methoxyxanthone and the less lipophilic compound was 1-methoxyxanthone. The acidity constants, at 25 ºC and with ionic strength of 0,15 mol dm-3 (NaCl), of mono- hydroxylated xanthones were determined by spetrophotometric titration, using the Hyperquad 2006 software [4]. The results have shown that the more acidic xanthones were 3-hydroxyxanthone and 4-hydroxyxanthone, and the less acidic were 1-hydroxyxanthone and 2-hydroxyxanthone. Mechanistic and structure-physicochemical values relationships were rationalized and can explain the obtained data for the different compounds in this study. [1] Pinto, M.M.M., Sousa, M.E. and Nascimento, M.S.J. (2005), Xanthone derivatives: New insights in biological activities, Current Medicinal Chemistry, 12 (21), 2517-2538. [2] Kerns, E.H. and Di L. (2003), Pharmaceutical Profiling in Drug Discovery, Drug Discovery Today, 8 (7), 316-323. [3] Castro, B., Gameiro, P., Lima, J.L.F.C., Matos, C. and Reis, S. (2001), Interaction of drugs with hexadecylphosphocholine micelles. Derivative spectroscopy, acid-base and solubility studies, Materials Science and Engineering: C, 18 (1-2), 71-78. [4] Gans, P., Sabatini, A. and Vacca, A. (1996), Investigation of equilibria in solution. Determination of equilibrium constants with the HYPERQUAD suite of programs, Talanta, 43 (10), 1739-1753.

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Dissemination of blaTEM-52 among Escherichia coli and Klebsiella pneumoniae strains is caused by spread of plasmids in Portugal

A. Pedrosa1, 2, A. Novais2, E. Machado1,3, R. Cantón2, T. M. Coque2*, L. Peixe1

1 Department of Microbiology, Faculty of Pharmacy, University of Porto, Portugal. 2 Hospital Universitario Ramón y Cajal, IMSALUD, Madrid, Spain 3 Faculty of Ciências da Saúde, Fernando Pessoa University, Porto, Portugal

TEM-52 was firstly described in 1996 and since then it has been sporadically identified in Europe, Korea and Canada among Enterobacteriaceae from both human and animals [1, 2]. Recently, an epidemic IncI1 plasmid containing blaTEM-52 was identified in France among Salmonella enterica community strains[2]. We analyzed the diversity of TEM-52 producing isolates from Portugal in order to understand the reasons for recent spread in our country. Twenty one TEM-52-producing isolates recovered from Portugal (n=19 Escherichia coli; n=2 Klebsiella pneumoniae) between 2002 and 2005 were studied. The isolates were obtained from hospitals (n=7), poultry (n=12) and sewage (n=2). Only one phenotype was included. Clonality was established by PFGE and E. coli phylogenetic groups by PCR as reported [3, 4]. ESBL identification was achieved by IEF, PCR and sequencing. Standard methods were used for conjugation assays as well as antibiotic susceptibility testing. bla location was addressed by hybridization of I-CeuI-digested DNA with an intragenic blaTEM-52 probe [5]. Plasmid characterisation was accomplished by the Barton’s method, identification of plasmid incompatibility groups by PCR, hybridization and sequencing, and analysis of RFLP patterns [6, 7]. The isolates studied were highly diverse and 21 PFGE-types were identified [3]. Most E. coli recovered from clinical isolates were assigned to phylogenetic group B1 [4]. E. coli recovered from poultry and sewage were identified as phylogroups A (4 A0 and 3 A1), B1 (n=2) and D (3 D1 and 1 D2). Although chromosomal location was identified in one strain, blaTEM-52 gene was mostly carried by two different plasmids of approximately 90kb and 30kb [1, 5]. The 90kb plasmids belong to the incompatibility groups I1 but the 30kb could not be typed by the PCR- typing method used. All the 30kb plasmids presented a unique RFLP pattern and most of the 90kb plasmids had the same RFLP pattern.

Wide and recent dissemination of blaTEM-52 among E. coli and K. pneumoniae strains from Portugal seems to be associated with dissemination of particular plasmids of 30 and 90kb among highly unrelated E. coli and K.pneunoniae strains. [1] Weill, F., Demartin, M., Fabre, L. and Grimont, P.A.D. (2004), Extended-Spectrum-β-lactamase (TEM- 52)-Producing Strains of Salmonella enterica of Various Serotypes Isolated in France, Journal of Clinical Microbiology, 42(7):3359-3362. [2] Cloeckaert, A., Praud, K., Doublet, B., Bertini, A., Carattoli, A., Butaye, P., Imberechts, H., Bertrand, ., Collard, J., Arlet, G. and Weill, F. (2007), Dissemination of an Extended-Spectrum-β-Lactamase blaTEM-52 Gene-Carrying IncI1 Plasmid in Various Salmonella enterica Serovars Isolated from Poultry and Humans in Belgium and France between 2001 and 2005, Antimicrobial Agents and Chemotherapy, 51(5):1872-1875. [3] Liu, S.-L., and Sanderson, K. E. (1992), A physical map of the Salmonella typhimurium LT2 genome made by using XbaI analysis, Journal of Bacteriology, 174(5):1662-1672. [4] Clemont, O., Bonacorsi, S. and Bingen, E. (2000), Rapid and Simple Determination of the Escherichia coli Phylogenetic group, Applied and Environmental Microbiology, 66(10):4555-4558. [5] Liu, S.-L., Hessen, A. and Sanderson, K. E. (1993), Genomic mapping with I-Ceu I, an intron-encoded endonuclease specific for genes for ribosomal RNA, in Samonella spp., Escherichia coli, and other bacteria, Proc. Natl. Acad. Sci. USA, 90:6874-6878. [6] Barton, B., Harding, G. and Zuccarelli, A. J. (1995), A General Method for Detecting and Sizing Plasmids, Analytical Biochemistry, 226:235-240. [7] Carattoli, A., Bertini, A., Villa, L., Falbo, V., Hopkins, K. L. and Threlfall, J. (2005), Identification of plasmids by PCR-based replicon typing, Journal of Microbiological Methods, 63:219-228.

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Synergism between adenosine A2A and tachykinin NK1 receptors facilitate [3H]-ACh release from myenteric neurons

I. Silva, C. Vieira, M. Duarte-Araújo & P. Correia-de-Sá

Laboratório de Farmacologia e Neurobiologia, Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar - Universidade do Porto (ICBAS-UP), Portugal.

Endogenous tachykinins, such as substance P (SP) and neurokinin A (NKA), are expressed in distinct neural pathways of the mammalian gut and have the potential to control both nerve and muscle activity [1]. In addition to inhibitory adenosine A1 receptors expressed on both cholinergic and tachykinergic myenteric neurons [2], activation of prejunctional A2A receptors facilitate acetylcholine (ACh) release from myenteric neurons [3]. Therefore, we decided to investigate the influence of endogenous adenosine on tachykinin-mediated facilitation of [3H]-ACh release from longitudinal muscle-myenteric plexus (LM-MP) preparations of the rat ileum. The experiments were performed at 37ºC on LM-MP preparations from control subjects (C) and from animals injected subcutaneously with capsaicin in the neonatal period (CAP). Neonatal capsaicin causes degeneration of peptidergic nerve fibres with loss of sensation and smooth muscle contractility. Procedures used for labeling the preparations and measuring evoked [3H]-ACh release were previously described [3]. 3 Facilitation of [ H]-ACh release caused by the selective A2A receptor agonist CGS 21680C (3 nM, 53±10%, n=5) was significantly (P<0.05) attenuated (10±9%, n=4) in CAP animals, but we failed to detect any changes on transmitter outflow with the inhibitory A1 receptor agonist (R-PIA, 300 nM). Adenosine deaminase (ADA, 0.5 U/ml), the enzyme that inactivates endogenous adenosine, was devoid of effect on CAP animals, while it 3 reduced evoked [ H]-ACh release by 31±6% (n=7) in C rats. The NK1 receptor agonist, 9 11 3 Sar ,Met(O2) -Substance P (s,m-SP, 100-300 nM), failed to modify [ H]-ACh release from both C and CAP rats. Co-application of s,m-SP (300 nM) plus CGS 21680C (3 nM) rehabilitated A2A facilitation (46±17%, n=6) on CAP animals. The selective NK1 antagonist, L-732,138 (20 nM) attenuated CGS 21680C (3 nM)-induced facilitation to 3 23±5% (n=4) in C rats. S,m-SP (300 nM) facilitated [ H]-ACh release when A2A receptors were activated by CGS 21680C (48±8%, n=4) or by increasing extracellular adenosine accumulation with EHNA (50 µM, 27±6%, n=5), which prevents deamination of the nucleoside. The adenylate cyclase activator, forskolin (3 µM), mimicked the enhancing effect of CGS 21680C on s,m-SP (300nM)-induced facilitation of [3H]-ACh release (31±6, n=4) in C rats. 3 Tonic facilitatory A2A receptor activation is impaired on CAP rats. Facilitation of [ H]- ACh release by NK1 receptor activation requires high levels of endogenous adenosine activating A2A receptors. Synergism between adenosine A2A and tachycinin NK1 receptors to facilitate [3H]-ACh release from myenteric neurons is probably mediated by an interaction at the adenylate cyclase / cyclic AMP transducing system level. [1] Holzer & Holzer-Petsche (1997) Pharmacol Ther.73, 173-217; [2] Broad et al. (1992) Am. J. Physiol. (Gastrointest. Liver Physiol.) 262, 525-531; [3] Duarte-Araújo et al. (2004) Br. J.Pharmacol. 141, 925-934. This work was supported by FCT (FEDER funding, POCTI/45549/FCB/2002) and PTDC/74462/CVT/2006 projects).

26

PARALLEL ORAL SESSIONS I – POS.I.2 WEDNESDAY, FEB. 20

I.2 - TERRITORY & NETWORK Elisa Keating & Laura Oliveira The knowledge-based economy as a hegemonic meta-object of urban centre renewal policies: the 201 case of Porto J. Queirós, V. Rodrigues "Massarelos Network": Spaces and times of inter-institutional articulation, in the perspective of an 34 "Educating Parish" Joana Lúcio 151 The Space of Intervals. The inner spaces of urban blocks: Space, Landscape, City Andreia Monteiro da Costa 120 Forms of Housing and Ways of Living in Porto's Contemporary Urban Territory André Mendes Faria 203 Interactive Application for Spatial Communication and Representation A.Vieira 186 Economical and social benefits of Astronomy André Filipe Soares Fernandes

The knowledge-based economy as a hegemonic meta-object of urban centre renewal policies: the case of Porto

J. Queirós1 and V. Rodrigues2

1 Institute of Sociology, Faculty of Arts, University of Porto, Portugal. 2 Institute of Sociology, Faculty of Arts, University of Porto, Portugal.

Abstract:

Inherently cultural, cities have always been, throughout History, privileged spaces of knowledge, creativeness, innovation and artistic effervescence. In the last few decades, following the profound economic, social and political transformations that have been characterizing cities, urban culture – considered here in its broadest sense – has gained renewed importance, especially in developed countries, as a central element in interurban competition. Relying on the economic convertibility of symbolic capital, contemporary urban policies choose knowledge and culture as two of their most important touchstones. The main goal is to assure the abovementioned conversion, by attracting new residents and tourists – especially the more affluent ones – and by reinforcing the urban centres’ attractiveness towards all kinds of investment fluxes. The complex strategies and discourses of urban centre renewal policies – and the way they increasingly mobilise the knowledge-based economy as their meta-object – is what we wish to explore with this paper. Focusing on the case of Porto, and having in mind the theoretical assessments of cultural political economy, we analyse the major guidelines of dominant tendencies in urban planning and management and reflect on the role of knowledge and culture in local urban centre renewal policies. By doing this, we will try to analyse how the international trend that takes the knowledge- based economy as a hegemonic meta-object of urban centre renewal policies is locally being reproduced.

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“Massarelos Network”: Spaces and times of inter-institutional articulation, in the perspective of an “Educating Parish”

Joana Lúcio1

1 PhD Student of Education Sciences, Faculty of Psychology and of Education Sciences, University of Porto, Portugal.

The investigation to which this report refers was developed in the context of the curricular probation year of the Education Sciences’ degree of licenciate, of the Faculty of Psychology and of Education Sciences of the University of Porto. One of our concerns was to ponder about the importance of the Seminar in the development of an intervention project. At the same time, we wanted to discuss the pertinence of the developed work and of the intervention project, considering the major objectives of the formation in Education Sciences and the role of the socio-educational and formation mediator (professional designation for Education Sciences’ graduates). In what concerns this last subject, we tried to discuss this socio-professional concept, giving special attention to the socio-political and professional opportunity that it produces and to the know-how, competencies and privileged targets of the mediator’s intervention. Our main objective was to build and analyze an interview database. These interviews were made to people responsible for local institutions, with the purpose of revealing various possibilities of understanding the importance of the local (the city, the parish, the neighborhood, the community) as a source of Education, giving that all aspects referring to the community life’s organization have an (educative) influence on the citizens’ lives. Theoretically speaking, we searched for the emergence of a structure similar to the Integrated Formative System [1], that we believe to allow the development of “joint spaces”: spatial-temporal contexts that mobilize the action of the people, the schools, the local associations, the autarchic power instances and the corporate sector, towards the prioritization of the public interests and needs. The analysis of the supra-referred database allowed the unveiling of the need for a Local Educative Project and a Local Education Council, both of which should orientate the local intervention on aspects such as: the democratization of the access to cultural assets, educating towards diversity, political and civic participation, and the assumption of public spaces as educative/educating spaces.

References: [1] Villar, M. B. C. (2001), A Cidade Educadora. Nova Perspectiva de Organização e Intervenção Municipal, Instituto Piaget, Lisboa.

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The Space of Intervals. The inner spaces of urban blocks: Space, Landscape, City

Andreia Monteiro da Costa1 Supervisor: Manuel Mendes, Architect

1 Faculty of Architecture, University of Porto, Portugal.

This research starts with the eyes of the traveller. A look vacant of history and pre-conceptions, which produces an act of seeing bringing together, in the same instant, the past and the present. The city grows further beyond the boundaries of institutions or masterplans and is without time restrictions. In the atemporality of the urban phenomenon, this work wanted to consider the city as a whole. The investigation uses Porto and the inner space of its urban blocks as the enquire territories for the research. Crossing ideas of several authors, such as Solà-Morales, Boeri, Joseph, Delgado or Lefebvre, the study made trough their ideas observations of the present. These were not made only through analysis, but the process of synthesis included both the involuntary memory of Proust and the atemporality of Sansot. The methodology also incorporated several other references from films and contemporary art, which helped both in the understanding and in the production of descriptive metaphors. This study produced a synthetic thinking, which, aiming for another legibility of the metropolitan form could be more similar to the more inclusive everyday experience of the city. Therefore, its intimate monuments build numerous personal networks of legibility, a kind of multiple ectoponymy, which contribute naturally to the hypertextuality of the urban phenomenon. In this differential of identities, Deleuze shifts the active figure from the point to the interval: “the interval takes all, the interval is substance”. So, in the city, the voids between different individual meanings become permanent constructors of new significances, and perpetuate the open essence of the urban organism. This study uses this notion of the interval to develop an idea of needed fluctuations and creative misunderstandings to show the poorness of a city’s legibility when based only on a closed common identity. In Porto, the inner spaces of the urban blocks appear as territories where the city escapes from its own institutional plan. In these territories, where rules are loose, the sense of place is built based on different degrees of formality and liberty through the appropriation of urban space. These define different modes of production of space that characterise the place and its physical and non physical relations with users. According to the openness of Heidegger: “Building, Dwelling, Thinking”; the performance of the discourse participates in this production of urban space by changing these relationships. In response to the lack of identity in our present urban spaces, this work tries to explore the creation of the possibility of permeability as done by contemporary art. This permeability allows spaces to better connect with their users, resulting in a stronger link between individuals and their city through an erotic process of possession. As Solà-Morales, this study claims the need of indefinite and unproductive spaces in a city. As other spaces in other cities, these inner spaces of the urban blocks might work as expressions of a kind of indetermination that is a factor of inclusion. Through their absences and possibilities, these spaces absorb memories, multiple narratives and performances, offering the indetermination of intervals towards more open and individual constructions of the city.

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Forms of Housing and Ways of Living in Porto’s Contemporary Urban Territory

André Mendes Faria 1

1 Architect, qualified by the Faculty of Architecture, University of Porto, Portugal.

The following abstract refers to my architectural degree final dissertation. [1] The main purpose of our research was to inquire and understand the actual state of collective housing in Portugal. Within Porto’s market supply, for the social, cultural and economical middle class standards demand, we have evaluated houses by their accuracy responding to the needs and uses of modern society. We have done a partial survey on Porto’s housing, commercialized between April and May 2005, corresponding to 16 complexes and nearly 1300 single apartments from where we extract four case-studies. An analytical system was developed in order to evaluate potential flexibility or limitations to the use of these domestic spaces, based on their geometry, environmental characteristics and functional distribution. Simultaneously, we attempted to identify some trends of contemporary life-styles that impact the occupation and use made of living space. Such aspects include increasing consumerism, cult of body, changes in gender roles and increasing leisure time. The confrontation between this and the analysis of the case-studies reveled some curious issues. Firstly, it shows that great majority of housing supply in Porto has the same functional structure and space dimensions, which does not match with the variety of lifestyles in contemporary society. Thus, we are limited to “a one size fit all” model. Moreover, this same structure could be found in market supply before democratic revolution in Portugal, meaning that 30 years of tremendous social evolution had little effect on house conception. Secondly, this research shows also that this stagnated functional structure used homogeneously for several years is based in a very rigid and hierarchical system that does not promote the flexibility, adaptability and durability a home should have. Furthermore, it reflects an obsolete system of values, that sometimes can even be contradictory, based on a traditional family with children where social life is assumed by one of the parents while domestic issues are assumed by the other. In addiction, we should mention the shallow technologic and constructive quality of housing in general and its high prices; such state of affairs is widely reported on the national media on a regular basis. Strangely, whether we are urban or suburban dwellers, part of a traditional or a monoparental family, are young or mature, conservative or liberal we all share one single living model with specific and limited efficiencies, meanings and symbolism. Definitively we are stuck with a one-size fit-all model. References: [1] FARIA, André Mendes, Caracterização do fogo na Habitação Plurifamiliar do Porto: introdução a um processo de estudo, Architectural degree final dissertation, Faculty of Architecture, University of Porto, 2005.

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Interactive Application for Spatial Communication and Representation

A.Vieira 1

1 CEAU, Faculty of Architecture, University of Porto, Portugal.

In the urban planning process, especially with the development of urban or architecture design, it’s imperative to provide an effective communication platform between the designers and the society. This bi-directional communication channel between the people more responsible for the design proposals and society in general, has better results when different types of representation and communication techniques are applied, since they make comprehension of the design clearer and less technical. Without a proper communication channel the participation of general society on the architectural design and development phases is reduced leading to a lack of important information and feedback from the users. In this context, the information technologies (IT), applied to the representation and communication of architectural and urban design, can offer the required tools to develop and provide an intuitive and easy-to-understand communication channel. This research work had as main objective the critical analysis of a significant set of different representation methods and technologies that can be applied to communicate and represent urban design and architectural proposals. Differences and advantages of each technique, either when used alone or in combination with others are compared. During the course of this work, coordinated by Pedro Neto1 a web application IASCR (Interactive Application for Spatial Communication and Representation) was developed for the investigation project CCRE (Spatial Communication and Representation Center) with the main objective of making more clear and efficient representation and communication of architectural and urban design projects. To test and gather the opinion and interaction of users were used two case studies: – E-Learning Café – an architectural project for the interior the interior space of a building at U. Porto that comprises a dynamic program for all the academic community and where university students, teachers and staff may exchange knowledge, share experiences and carry out a wide range of activities; – Corredor da Memória - an urban design renovation project that upgrades an important historical area of the city of Porto, considered world heritage. The paper presents first a concise literature review and computer visualization techniques to communicate the design. It then proceeds to analyze the results of the questionnaire and the qualitative provide some positive information about how people perceived and judged both the content (proposed design) and the form (representations methods and techniques used for communicate).

1 Pedro Leão Neto -FAUP

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Economical and social benefits of Astronomy

André Filipe Soares Fernandes 1

1 Student of the department of Applied Mathematics, Faculty of Sciences, University of Porto, Portugal.

I'm taking the Master degree of the Faculty of Sciences in teaching of astronomy. My goal for the master thesis will be to research the economic & social benefits of Astronomy. Despite the obvious historical contributions of Astronomy to human civilization as well as the current externalities of space activities, there is not much organized information about them. This study intends to analyze the effects of Astronomy in our society. At this point, the following question arises: which effects? The study may take into account several perspectives. It may focus on the scientific and educational side of Astronomy or on the positive externalities of this science (technology, employment, growth, etc). Alternatively, one may think in Astronomy as the motivation to space exploration. On this point of view, it would be useful to understand the economic impact of space activities trough, for example, technology transfer or the services it generates and feeds (ICT, Telecommunications, Environment monitoring, Security, Civil Protection, Positioning, etc). Presently, the main challenge is to choose a perspective in order to focus the research. In the end, the study should be able to promote an objective discussion about the value of Astronomy and Space activities. If my application for IJUP 08 is successful, I will present a brief compilation of the facts already found in my early research. The thesis is supervised by the teacher Catarina Lobo from the Department of Applied Mathematics, Faculty of Sciences, University of Porto, Portugal.

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PARALLEL ORAL SESSIONS I – POS.I.3 WEDNESDAY, FEB. 20

I.3 - MODELLING & AUTOMATIC SYSTEMS Sofia Rodrigues & Filipe Abrantes Estimation of postmortem interval based on the determination of potassium in vitreous humor by an 83 automatic system Ana I. Pereira, Ana M. Santos, Marieta L.C. Passos, J. Rodrigo Santos, S. Frazão, Agostinho J.C.

Santos, M. Lúcia M.F.S. Saraiva and José L.F.C. Lima 111 The dynamical state of galaxies: from 2D to 6D P. Mondim, J. Brinchmann Development of an automatic multi-pumping flow system for the spectrophotometric determination 121 of trimipramine D. Ribeiro, J. Prior, J. Santos, J. Lima 150 Anti-de Sitter space and the stability of scalar field C. A. dos Santos Guedes Accessibility and science communication – new directions and guidelines for Moodle “Estaleiro da 176 Ciência” N.Regadas, J. Santos, B.Giesteira 192 Quantum Vacuum Energy in General Relativity (The Casimir Effect) M. Barbosa

Estimation of postmortem interval based on the determination of potassium in vitreous humor by an automatic system

Ana I. Pereira1, Ana M. Santos1, Marieta L. C. Passos1, J. Rodrigo Santos1, S. Frazão2, Agostinho J.C. Santos2, M. Lúcia M. F. S. Saraiva1 and José L. F. C. Lima1

1 REQUIMTE, Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Portugal. 2 Serviço de Medicina Legal, Faculdade de Medicina, Universidade do Porto, Portugal. Serviço de Patologia Forense, Instituto Nacional de Medicina Legal, I.P. – Delegação Norte.

The estimation of the time since death known as postmortem interval (PMI) is a main issue in the field of forensic science and legal medicine and one of the most difficult and challenging problem for forensic pathologists [1, 2]. In the present work is then proposed the estimation of the PMI based on postmortem potassium levels in vitreous humor. Vitreous humor has been regarded as the ideal extracellular fluid to measure the postmortem release of intracellular potassium (mainly from the retina) consequent to the energy breakdown and the related cessation of active transport and selective membrane permeability [3]. The quantification of potassium levels in vitreous humor will be performed using an automatic flow based method. Flow techniques are attractive automation tools as they allow the implementation of low cost, fast, reliable and reproducible methodologies. In this work a tubular potassium ion-selective electrode will be incorporated in the flow manifold as detector. The tubular electrode consists of a sensing element made up by a polyvinylchloride (PVC) membrane coating a tube of conductive material which serves both as support for the PVC film and as electric contact [4]. The use of this kind of detection provides a high sensitivity, low chemical and mechanical interferences, a short response time and a high reproducibility. This methodology will constitute a way of analyzing a huge number of samples rapidly in order to get significant statistical results to establish a mathematical relation between this parameter and the PMI. References: [1] McDowall, K. L., Lenihan, D. V., Busuttil, A. and Glasby, M. A. (1998), The use of absolute refractory period in the estimation of early postmortem interval, Forensic Sci Int.,91 (3), 163-170. [2] Henssge, C. and Madea, B. (2007), Estimation of the time since death, Forensic Sci Int.,165(2- 3), 182-184. [3] Tagliaro, F., Manetto, G., Cittadini, F. Marchetti, D., Bortolotti, F. and Marigo, M. (1999) Capillary zone electrophoresis of potassium in human vitreous humour: validation of a new method, Journal of Chromatography B, 733, 273-279. [4] Almeida, M. I. G. S., Segundo, M. A., Lima, J. L. F. C. and Rangel, A. O. S. S, (2006) Potentiometric multi-syringe flow injection for determination of exchangeable potassium in soils with in-line extraction, Microchemical Journal, 83, 75-80. Acknowledgements: Marieta L. C. Passos thanks Fundação para a Ciência e a Tecnologia and FSE (III Quadro Comunitário) for the Ph.D. grant (SFRH / BD / 22752 / 2005). Authors thanks Reitoria da Universidade do Porto the Project Investigação Científica na Pré-Graduação (2007).

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The dynamical state of galaxies: from 2D to 6D

P. Mondim 1, J. Brinchmann2

1 Department of Applied Mathematics, Faculty of Science, University of Porto, Portugal. Centre for Astrophysics of University of Porto, Portugal 2 Leiden University, The Netherlands. Centre for Astrophysics of University of Porto, Portugal

The goal of this work is to investigate the relationship between the dynamical state of a galaxy (which belongs to a 6D phase-space, and can only be known with information regarding the velocity and position of every galaxy's particles) and the information available from observations: it's 2D velocity field. This goal is pursued by numerically calculating the Tensor Virial Theorem [1] quantities to study the phase-space structure of galaxies mergers' simulations and contrasting them with synthetic observations of those galaxies. An effective way of calculating the kinetic energy associated with movement that can be regarded as “ordered” and the kinetic energy associated with movement that can be regarded as “random” has been devised. Using this tool to study the phase-space structure of merging galaxies, we were able to verify how ordered- and random-motion evolve during the interaction of the galaxies. The ratio of ordered-motion to random-motion of the system's particles has been found to peak early after the beginning of the interaction, presenting a strong decrease thereafter and, finally, reaching a steady level, lower than the initial value. Galaxies of different morphological types have been found to provide somewhat different evolutions for the quantities that trace the ordered- and random- motion. Work is now underway to create synthetic observations of those same mergers, in order to make possible a comparison of the “observed” and “real” physical quantities. This may lead, ultimately, to the establishment of innovative methods of non-parametric classification of velocity fields. Such methods would be specially valuable for high- redshift galaxies, for which the known methods fail for requiring high signal-to-noise ratios. Thus, so far, the main conclusions for the current work are the establishment of typical evolutions for the ordered- and random-motion of a galaxy's particles during a merger event and the establishment of different trends for the evolution of those quantities in galaxies of different morphological types. Further work is still underway.

References: [1] Binney, J. and Tremaine, S. (1994), Galactic Dynamics, Princeton University Press.

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Development of an automatic multi-pumping flow system for the spectrophotometric determination of trimipramine

D. Ribeiro 1, J. Prior 1, J. Santos1 and J. Lima1

1 Requimte, Department of Physical-Chemistry, Faculty of Pharmacy, University of Porto, Portugal.

Trimipramine maleate, is a tricyclic antidepressant agent, that belongs to the dibenzoazepine class, with an anxiety-reducing sedative activity. The intensive therapeutical utilisation of dibenzoazepine derivates increases the need to improve or develop new methods for their determination in body fluids and in pharmaceutical preparations [1]. In the present work, an automatic methodology of flow analysis exploiting the multi- pumping [2] concept was developed for the spectrophotometric determination of trimipramine in commercially available pharmaceutical formulations. The trimipramine determination was based on the reaction of the drug with ammonium monovanadate reagent in acidic medium yielding a coloured compound with a maximum of absorbance at 620 nm [3]. The improved flow mixing conditions during sample and reagents insertion and transport, due to the chaotic movement of the solutions originated by the pulsed flow characteristic of multi-pumping systems, assured a fast reaction zone homogenization in a reduced residence time, which was particularly advantageous for carrying out analytical determinations that involved highly viscous solutions, as is the case of the sulphuric acid solution used in the determination of trimipramine, without impairing the sampling rate. A linear working range for trimipramine concentrations of up to 50 mg L-1 (r = 0.9998; n = 6) was obtained and the detection limit was about 1.15 mg L-1. The sampling rate was approximately 50 determinations per hour. The obtained results were in agreement with those furnished by the reference procedure [4], with relative deviations lower then 4.7%. The proposed methodology allowed the rapid quantification of trimipramine, which could represent an advantageous alternative for the pharmaceutical control at industrial level, and additionally, required low quantities of reagents and produced reduced volumes of residues. References: [1] Baldessarini, R.J. (2001), Drugs and the treatment of psychiatric disorders. Depression and anxiety disorders, in Gilman, A.G., Hardman, J.G., and Limbird, L.E. (Eds.) The pharmacological basis of therapeutics, 10th ed., New York: McGraw Hill Co., pp. 447-483. [2] Lapa, R.A.S., Lima, J.L.F.C., Reis, B.F., Santos, J.L.M., Zagatto, E.A.G. (2002), Multi- pumping in flow analysis: concepts, instrumentation, potentialities, Anal. Chim. Acta, 466, 125- 132. [3] Misiuk, W. (2000), Spectrophotometry assay of imipramine and desipramine using ammonium metavanadate and its application to pharmaceutical preparations, J. Pharm. Biomed. Anal., 22, 189- 196. [4] Trimipramine Maleate monograph, tablets (2005), in British Pharmacopoeia, vol. III, London: The Stationary Office, pp. 2853.

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Anti-de Sitter space and the stability of scalar field

C. A. dos Santos Guedes1

1 Department of Physics, Faculty of Sciences, University of Porto, Portugal.

This work grew up as an introduction to what I have been doing in my final undergraduate thesis, a purely theoretical physics research based on what is called the AdS/CFT duality, under the supervision of Dr. Miguel da Sousa Costa. AdS stands for anti-de Sitter space which is a solution to Einstein’s field equations of general relativity in empty space with negative cosmological constant. Being a maximally symmetric space, it has been an excellent model to investigate questions of principle related to the quantization of fields propagating on curved background, the interaction with the gravitational field and the issues related to its lack of global hyperbolicity. Firstly, I focused on the geometric properties of anti-de Sitter and understood how subtle this space is, where time warps in itself and light reaches the spatial infinity in finite time. Moreover the symmetries, or the group to which this space is invariant, induce a natural behavior on its boundary that is conformally invariant. CFT above denotes conformal field theory. Afterwards, I delved into the propagation of scalar field. The scalar field corresponds in real world to what we can think of spinless particles (spin is an intrinsic property of particles like mass and charge). For us to have particles they must be stable to the extent that we can manipulate them in a consistent way, i.e., be able to do quantum field theory. This part was based on work done almost thirty years ago [1-3], which led to the conclusion that the scalar field is stable in any dimension for an AdS fixed background. Because the mathematical apparatus and concepts used are far from being trivial, and the audience is a very general one, the main part of the presentation will be concerned with very general ideas about what General Relativity, Quantum Mechanics and Quantum Field Theory is all about and why it is important to think of them from a unified point of view. If time permits, further questions specific to this work could be addressed.

References: [1] Breitenlohner, P. and Freedman, D. Z. (1982), Positive Energy in Anti-de Sitter Background and Gauged Extended Supergravity, Phys. Lett. B 115. [2] Breitenlohner, P. and Freedman, D. Z. (1982), Stability in Gauged Extended Supergravity, Ann. Phys. 144. [3] Mezincescu, L., and Townsend, P. K. (1985), Stability at a Local Maximum in Higher Dimensional Anti-de Sitter Space and Applications to Supergravity, Ann. Phys. 160.

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Accessibility and science communication – new directions and guidelines for Moodle “Estaleiro da Ciência”

N.Regadas 1,2, J. Santos2, B.Giesteira3

1 Jornalismo e Ciências da Comunicação, Faculdade de Letras, Universidade do Porto, Portugal. 2 IBMC (Instituto de Biologia Molecular e Celular – Universidade do Porto). 3 Faculdade de Belas Artes, Universidade do Porto, Portugal.

Science communication on a web context raises several accessibility and usability questions that us, as communicators, should recognize and exceed. Because Moodle has one of the highest worldwide implementation ratings on e-learning and b-learning platforms, the adoption of good web usability and accessibility practices, at all levels, becomes mandatory. “Estaleiro da Ciência - Oficinas” has a different approach on science teaching and learning, aiming to be a model for future b-learning applications. Having W3C guidelines for a starter and following Nick Freear and Chetz Colwell [1] accessibility report on Moodle 1.6 version, as well as other documents, we examined the 1.7 version to see if it was according to those guidelines. Our main goal is to format the "Estaleiro da Ciência" Moodle, so that it is in total conformity with the first accessibility level, proposed by WAI of the W3C. The IBMC “Estaleiro da Ciência – Oficinas” cluster was subjected to automatic and manual accessibility evaluations; for usability evaluations Lavery’s [2] modified structure of Nielsen’s Heuristics was used among an empiric observation method with five users; the four months of empiric experience working with the platform, and the discussions on Moodle forums were also considered for this study case. Our objective is to bring the programmer’s and the designer’s world closer together. Although some changes are only possible if php programming is used, others can be done resorting to CSS and internal HTML editors. Labels and HTML blocks are useful tools to develop and improve accessibility features. Despite the good accessibility results of Moodle 1.7 version, in compliance with WAI first level of priority, an additional effort is being made to make it compliant with other W3C standards. Special attention was given to navigational elements as they are one of the most important elements on a system like this. Our results revealed that usability can be improved virtually without any programming involved. Tests revealed that too many side blocks, links and options lead to user’s frustration. This variable is controllable if changes are made to the course’s layout. Users must be fully concentrated on the contents, instead of feeling lost with such a vast amount of information. Moodle’s standard version is not very user friendly, even though it’s not very difficult to learn how to operate it. The main idea is that technology has to adapt to our needs and not the other way. Although this is a study case, all drawn conclusions and recommendations can be implemented to similar structures. This project is still ongoing, and has a huge potential of applicability and future work perspectives. References: [1] Freear, Nick and Colwell, Chetz., Improving the Accessibility of Moodle, Experiences, guidelines and the road ahead, The Open University, 2006 [Acessed 25th March 2007] Available:http://moodlemoot.org/file.php/3/presentation_materials/nick.f_chetz.c_OU/MoodleMoo t_slides_ndf_cc_FINAL_2.pdf [2] Heuristic Evaluation was originally proposed by Nielsen and Molich (Nielsen and Molich, 1990). The structure was, later on, modified by Nielsen (Nielsen, 1994).

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Quantum Vacuum Energy in General Relativity (The Casimir Effect)

M. Barbosa1

1 Department of Physics, Faculty of Science, University of Porto, Portugal.

My work is about the calculation of the zero-point energy (or vacuum energy) of a scalar field in a curved space, which exists by a phenomena known as Casimir effect. In 1948, Casimir showed that the fluctuations of the electromagnetic field in vacuum, between to parallel conductive plates, is not zero and gives rise to an attractive force between them. It’s a very weak force but perfectly measurable. Applying Casimir’s calculation method to general relativity, we can verify that when we have a scalar field (that can represent a particle, for example) in a curved space, if we consider its small fluctuations, we will obtain a non-zero vacuum energy. A lot of research work is being done to understand this non-zero vacuum energy and its relationship with the deformation of the space-time. This is a theoretical work, involving hard mathematical concepts. As we will have a very general audience, I will not focus my presentation in the expressions and results I found. Instead, I will try to give a basic point of view about what is a curved space, vacuum energy, etc., and what kind of phenomena we can get with its connections. This will put the audience closer to a big Physics’ topic - The General Relativity. Resuming, I will try not to show just complicated equations that the most part of the audience wouldn’t understand, but I will try to make them have a simple and intuitive idea about the physical concepts used in my work.

References: [1] L. H. Ford, Phys, Rev. D 11, 3370 (1975) [2] L. H. Ford, Phys. Rev. D 14, 3305 [3] N. D. Birrell, P. C. W. Davies, Quantum fields in curved space, Cambridge University Press (1999) [4] K. A. Milton, The Casimir Effect, World Scientific (2001)

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PARALLEL ORAL SESSIONS II – POS.II.1 THURSDAY, FEB. 21

II.1 - FOOD & HEALTH Maria Paula Santos & Jorge Teixeira 60 Microplate high-throughput methodology for determination of Folin-Ciocalteu index F. Santos, L. M. Magalhães, M. A. Segundo, S. Reis, J. L. F. C. Lima 100 Detection of authorized genetically modified maize events: Participation in an inter-laboratorial study J. Rocha, I. Mafra, M. Carvalho, J.A. Amaral, M.B.P.P. Oliveira 101 Detection of genetically modified soybean in foodstuffs: Participation in an inter-laboratorial study M. Carvalho, I. Mafra, J. Rocha, M.B.P.P. Oliveira HPLC separation and quantification of Catechins in green tea home prepared: comparison with 160 commercial soft drinks C. Petisca, A. Melo, O. Pinho, I.M.P.L.V.O. Ferreira 182 Product Engineering - Application of vanillin in alimentary food paste industry A. Vasconcelos, B. Moura, L. Almeida, S. Couto 200 Development of a coating barrier for cork stoppers for their use in spirituous drinks F. Oliveira, S. Pontes, M. Cabral, A. Mendes

Microplate high-throughput methodology for determination of Folin-Ciocalteu index

F. Santos, L. M. Magalhães, M. A. Segundo, S. Reis and J. L. F. C. Lima

REQUIMTE, Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal Email: [email protected]

The interest in antioxidant nutrients has increased due to their protective effects against free radical-induced reactions [1]. In this context, the Folin-Ciocalteu assay [2] has been proposed recently as a standard method for measurement of antioxidant capacity of food products and dietary supplements in routine quality control [3]. Therefore, the main goal of this work is the development of a simple, fast and reliable analytical methodology for the determination of Folin-Ciocalteu index. To achieve this objective, the conventional methodology was adapted to a 96-well microplate format. The reaction volume was then reduced 100 times, from 20 mL to 0.2 mL. Furthermore, the reaction conditions were thoroughly studied, namely the pH of the reaction media. It was observed that an increase of pH from 10 (usual value applied, provided by a carbonate buffer) to values around 12-13 changed the reaction kinetics dramatically. In fact it was possible to decrease the reaction time from 2 hours to 3-5 minutes. The developed methodology was applied to the determination in several types of food products, using gallic acid as standard compound. The Folin-Ciocalteu index values obtained for wines, beers and juices were comparable to those obtained from the previous batch method, with a significant improvement in the reagent consumption, effluent produced per assay and determination throughput. These characteristics constitute a step forward the implementation of greener procedures in the Analytical Chemistry area.

References: [1] Halliwell, B., Murcia, M.A., Chirico, S., and Aruoma, O.I. (1995), Free-radicals and antioxidants in food and in vivo –what they do and how they work, Critical Reviews in Food Science and Nutrition, 35 (1-2), 7-20. [2] Singleton, V.L., Orthofer, R., and Lamuela-Raventós, R.M. (1999), Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent, Methods in Enzymology, 299, 152-178. [3] Prior, R.L., Wu, X., and Schaich, K. (2005), Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements, Journal of Agricultural and Food Chemistry, 53 (10), 4290-4302.

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Detection of authorized genetically modified maize events: Participation in an inter-laboratorial study

J. Rocha 1,2, I. Mafra1, M. Carvalho1, J.A. Amaral1, M.B.P.P. Oliveira1

1 REQUIMTE-Laboratory of Bromatology, Faculty of Pharmacy, University of Porto, Portugal. 2 Faculty of Sciences, University of Porto, Portugal.

The soybean and maize are the most important genetically modified (GM) crops (57% and 25% of global biotechnological planted area, respectively) [1]. The need to monitor and verify the presence of biotechnology-derived material in food products demands analytical methods able to detect, to identify and to quantify either the introduced DNA or the expressed protein(s). The interest of USDA/GIPSA Proficiency Program is to perform accurate, reliable, and reproducible testing on biotechnology-derived events. The objective of this Program is to detect and quantify GM maize and soybean flours by means of DNA and/or protein based methods to verify the performance of several laboratories of individual organizations. The aim of the present work was the detection of GM maize events of six ground maize samples supplied by the Proficiency Program in two periods. The samples were received with the information of 11 specific GM events ranging from 0 to 5%. DNA molecules were the target compounds for GMO detection due to the higher sensitivity of DNA-based methods and to their higher stability compared to proteins. DNA was extracted by two different methods [2]: CTAB and Wizard. Yield and purity of DNA extracts were assessed by spectrophotometry, while amplifiability was evaluated by PCR targeting the invertase gene. DNA extracts were amplified by two polymerase chain reaction (PCR) techniques: qualitative PCR and real-time quantitative PCR. Several qualitative PCR techniques were performed to screen the 35S promoter sequence and to detect six GM events: Bt11, MON810, E176, GA21, NK603 and MON863. A real-time PCR assay with TaqMan probes was performed to quantify MON810. The results received by the first period of the Program showed good performance of the Laboratory of Bromotology to screen GM events by targeting the 35S promoter and by the detection of Bt11, E176 and MON810. Concerning the second period of the Program, where the other events were tested for the first time, the results are not yet available.

References: [1] James, C. (2006), Global Status of Commercialized Biotech/GM Crops:2006, ISSAA Brief No. 35, New York, Ithaca: ISSAA. [2] Mafra, I., Silva, S.A., Moreira, E.J.M.O., Ferreira da Silva, C.S., Oliveira, M.B.P.P. Comparative study of DNA extraction methods for soybean derived food products, Food Control (submitted).

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Detection of genetically modified soybean in foodstuffs: Participation in an inter-laboratorial study

M. Carvalho1, I. Mafra1, J. Rocha 1,2, M.B.P.P. Oliveira1

1 REQUIMTE-Laboratory of Bromatology, Faculty of Pharmacy, University of Porto, Portugal. 2 Faculty of Sciences, University of Porto, Portugal.

In last ten years, the agriculture has been revolutionized by the introduction of genetically modified organisms (GMO). The soybean is one of the most important genetically modified crops, totalizing 83.8 milions of planted hectares, which correspond to 57% of biotechnological planted area [1].The acceptance of GMO by consumers is controversial, and concerns about their safety persist among public opinion. The EU legislations demand the labelling of food products containing more than 0.9 % of GM material (Regulation (EC) N.º 1829/2003). The interest of USDA/GIPSA Proficiency Program is to perform accurate, reliable, and reproducible testing on biotechnology-derived events. The objective of this Program is to detect and quantify GM maize and soybean flours by means of DNA and/or protein based methods to verify the performance of several laboratories of individual organizations. The aim of the present work was the detection of GM soybean of three ground soybean samples supplied by the Proficiency Program in two periods. The samples were received with the information of one specific GM event ranging from 0 to 5%. The most accepted techniques for GMO detection rely on DNA based methods, namely polymerase chain reaction (PCR) or real-time quantitative PCR techniques due to their high specificity and sensitivity. Thus, the PCR techniques were the choice for the present study. The isolation of DNA from soybean flours was carried by using two methods: the CTAB and the Wizard [2]. Yield and purity of DNA extracts were assessed by spectrophotometry, while amplifiability was evaluated by PCR targeting the lectin gene. Two types of PCR assays were developed and used: convetional PCR assays for detection of Roundup Ready (RR) soybean and real-time PCR assays for quantitative purposes. The results received by the first period of the Program showed good performance of the Laboratory of Bromotology to screen GMO by targeting the 35S promoter and by the qualitative detection of RR soybean. Concerning the second period of the Program, the results of the three samples analysed by real-time PCR with TaqMan probes were: 0.8%, 2.5% and 0.2% of GMO. The report for the second period is not yet available to conclude about the performance of the present work.

References: [1] James, C. (2006), Global Status of Commercialized Biotech/GM Crops:2006, ISSAA Brief No. 35, New York, Ithaca: ISSAA. [2] Mafra, I., Silva, S.A., Moreira, E.J.M.O., Ferreira da Silva, C.S., Oliveira, M.B.P.P. Comparative study of DNA extraction methods for soybean derived food products, Food Control (submitted).

44

HPLC separation and quantification of Catechins in green tea home prepared: comparison with commercial soft drinks

C. Petisca1, A. Melo2, O. Pinho1,2, I.M.P.L.V.O. Ferreira2

1Faculdade de Ciências da Nutrição e Alimentação, Universidade do Porto, Portugal 2REQUIMTE – Departamento de Bromatologia, Faculdade de Farmácia, Universidade do Porto, Portugal

Tea is produced from the leaves of the plant Camelia sinensis. Tea composition varies with climate, season, horticultural practices, variety, type of soil and the age of the leaves [1]. The high concentration of catechins and theaflavins in tea, coupled with its broad consumption, make tea a significant dietary source of these flavonoids. These compounds are potent natural antioxidants. Epidemiological and experimental evidence have highlighted potential therapeutic and health-protective activities of tea catechins and related derivatives [2]. Green tea samples were prepared using an aqueous extraction procedure which simulated usual brewing conditions for a cup of tea. A precisely known amount of green tea leaves was steeped at different temperatures (100°C, 95°C, 90°C, 85°C, 80°C, 75°C and 70°C) for 5 and 10 minutes respectively and compared with ideal conditions for catechins extraction (80°C during 80min) to adquire maximum concentration of catechins. The amount of catechins present in commercial soft drinks containing green tea was also studied and the results were compared. Separation and quantification of catechins was performed by reverse-phase HPLC with Diode array detection. The samples were filtered through a 0,45μm polyester filter before analysis. Evaluation of time/temperature relation in green tea samples prepared using home conditions, showed that samples prepared at 100°C for 10minutes, contained more catechins than the other time/temperature relations evaluated, but has about 10 times less catechins than the ideal conditions for catechins extraction. Comparison with the levels of catechins in comercial soft drinks revelead that “Lipton Linea” infusion contained higher amounts of catechins. However, this infusion presented less galic acid when compared with tea at 100°C for 10min. Cold “Lipton Linea” also contained high levels of catechins. The others comercial soft drinks analized presented lower amounts of catechins. In conclusion the ideal time/temperature to extract the highest amount of catechins is 80°C for 80min, but , despite being the most accurate way, people at home don’t take so much time to prepare tea. When comparing catechin content present in tea prepared at home conditions, we concluded that some comercial soft drinks have more amount of catechins.

References: [1] Yang, D.J., Hwang, L.S. and Lin, J.T. (2007), Effects of different steeping methods and storage on caffeine, catechins and gallic acid in bag tea infusions, Journal of Chromatography, 1156 (1-2), 312-320. [2] Neilson, A.P., Green, R.J., Wood K.V. and Ferruzzi, M.G. (2006), High-throughput analysis of catechins and theaflavins by high performance liquid chromatography with diode array detection, Journal of Chromatography, 1132 (1-2), 132-140.

45

Product Engineering – Application of vanillin in alimentary food paste industry

A. Vasconcelos, B. Moura, L. Almeida and S. Couto

Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal.

Vanillin is the chemical substance accountable for the vanilla aroma and it is also one of the most used aromatic compounds in alimentary industry. This compound can be obtained either by natural way or synthetically. In the sphere of Product Engineering an innovation product has been developed in alimentary pastes industry - vermicelli vanilla cinnamon, whose benchmark [1] would be the common vermicelli. The production at a pilot scale showed that it would really be an attractive innovation and the economical plan revealed that this would be practicable in economical terms. The experiences were realized in a pilot scale, by means of mixing about 100g of semolina of hard wheat [2] (which is the unique ingredient common to benchmark[1]) and water, with 1 – 3g of cinnamon and different volumes of vanillin solution (between 1ml and 10ml). The obtained paste was pressed and cut in an extrusion machine. At last, it was dried in a stove at about 50°C. Supposing that our product should be made in a well – known enterprise in the market, we made an economical and marketing analysis. With the obtain results, it was made the design of a package (Fig.1) that should arise the public interest in general. After the experience in a pilot scale had been realized, we concluded that vermicelli with about 3 – 5 ml of vanilla solution per 100g of semolina would be the tastiest. The market study has showed that this new concept of vermicelli should be well accepted by the public, and the economical analysis revealed that the innovation should be profitable[3].

References: [1] Milaneza, www.milaneza.pt, accessed in 28th November 2007. [2] www.ama-massas.com/massasfr.htm, accessed in 23th November 2007. [3] Vasconcelos A., Moura B., Almeida L., Couto S., Vanilina – da Inovação ao Design, internal full report for the discipline of Product Engineering of the Integrated Master’s in Chemical Engineering from FEUP, 2007.

46

Development of a coating barrier for cork stoppers for their use in spirituous drinks

F. Oliveira 1, S. Pontes 1, M. Cabral 2 and A. Mendes 3

1 The Network for Competence in Polymers, Faculty of Engineering, University of Porto, Portugal 2 Amorim & Irmãos Company, Mozelos, Portugal 3 Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal.

Presently, spirituous drinks such as vodka are not bottled using cork stoppers, despite the great appetence of the market for the noble product. The reason for that is the bleeding of natural colorants present in the cork stoppers to the white drink, making it looks like old conhaque. Two approaches could in principle be followed to make the cork stoppers compatible with spirituous drinks. The first one is the extraction of these colorant compounds leaving a colorant-less cork stopper. The other approach concerns the development of a food approval and highly flexible transparent coating, impermeable to the colorant compounds and chemically stable. At first glance none of these approaches seem to be easy to accomplish. This is indeed true and there is presently no good solution for this challenge. The extraction of the natural colorants, namely tannins, from cork stoppers proved only to delay the color bleeding, even when the extraction was applied to fine cork granules. Different extracting solvents were employed as well as supercritical carbon dioxide. Some authors proposed the use of polymers such as silicon [1, 2]. Despite the great permeability of this polymer, it is food approval, elastic and chemically resistant. It also seems to retain, sorb, the bleeding colorants from the cork stopper. However, silicon is also not able than delay the bleeding of colorant to the beverage. The authors tool advantage of the polarity of the cork stoppers colorants and developed a coating system considering two layers. The first layer fences the colorants to leave, due to its charge, while the second layer provides mechanical resistance to the coating. Both polymers are commercially available and are applied by dip coating. The final coating system is food approval, very elastic, chemically stable and gas permeable. Moreover, it is tasteless, transparent and visually attractive. Presently, a new monomer is being investigated that for developing a polymer that offers complete barrier properties with a single layer.

References: [1] WATKINS, S. (1997), Coated cork stopper, WO9711894; [2] LUMIA, G., PERRE, C., ARACIL, J. (2001), Method for treating and extracting cork organic compounds, with a dense fluid under pressure, WO0123155;

47

PARALLEL ORAL SESSIONS II – POS.II.2 THURSDAY, FEB. 21

II.2 - HOUSE & LIVING Gonçalo Furtado & Albino Lima 45 The Architect's House Joana Ferraz 205 Organic living | Weekend house in Castelo de Paiva P. Geraldes Santos, N. Brandão Costa 171 From RCR experience to a project of a small house in the mountain F. Costa Leite, N. Brandão Costa Casa Comum [common/collective/communitarian house/home] Architectures for an Intermediary 122 Space Nuno Travasso 43 The Courtyard and Siza's Houses H. Henriques 155 The silence and the sky: seclusion and architecture. Four courtyard-houses. A. Silva Fernandes, C. Machado

The Architect’s House

Joana Ferraz 1

1 Architecture graduate, Faculty of Architecture, University of Porto, Portugal.

The architect’s own house is a recurring subject in architecture publications. The works of domestic architecture are analyzed by their characteristics, the presence of the author, influences, design process, etc. But what happens when the relation client/architect is altered? What happens when architect and client are one and the same person? Which are the main differences between the current outputs and their own house? What to state when the subject of study is “The Architect’s House"[1]? My interest, in this particular subject, came from a casual reading of a Spanish book regarding the same theme – La casa del arquitecto [2]. However, this study employs bibliographical references associated which such distinctive areas as philosophy and sociology: authors as Foucault or Heidegger, architecture historians as Joseph Rykwert, or even writers, as T.S.Elliot, carried out active parts. The investigation’s net is sewed, fundamentally, within the Portuguese 20th century, unifying an assembly of apparently divergent works but after all with a common background. Besides representing epochs and tangible architectonic tendencies, through them one would be able to reproduce the historical line of domestic architecture in the past century: they reproduce the architect’s personal history, reflecting ideals and choices, architectonic principles and aspired lifestyle. To understand the spirit and variables of the conception and execution of the architect’s own house, preceding the case studies and subsequent national contextualization, some chapters specifically explore themes such as the origin of the creative act, the architect’s adaptation and proximity to the creative individual/author’s task, the performance of the author’s role in society, and the confrontation and affinity of the individual with the collective. Subsequently, articulations were established between such concepts as architect/individual/author and house/residence/home and several of its possible interpretations, as well as the different varieties of architect’s own houses including its correlation with heterotopies. In perspective this is a fertile field with countless ramifications that may be subject to future inquiries such as establishing parallelisms between Portuguese cases and those of foreign architects, or a more detailed approach on the variability between own houses and remaining works of the same author. “The Architect’s House” is, ultimately, an architectural object that permits others to perceive, with clarity, the heart and the contradictions in the architecture of a specific author.

References: [1] Ferraz, Joana, (2007), The Architect’s House, Graduation Thesis in Architecture, Faculty of Architecture of Porto University [Ferraz, Joana, (2007), A Casa do Arquitecto, Prova Final de Licenciatura em Arquitectura, Faculdade de Arquitectura da Universidade do Porto, Porto.] [2] Zabalbeascoa, Anatxu, (2000), La casa del arquitecto; Gustavo Gili, Barcelona.

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Organic living | Weekend house in Castelo de Paiva

P. Geraldes Santos 1, N. Brandão Costa2

1 Faculty of Architecture, University of Porto, Portugal. 1 Faculty of Architecture, University of Porto, Portugal.

The present work appeared in the context of a bid of a project for a house in a rural area near the river Douro, more precisely in the village of Castelo de Paiva, located in the north of Portugal. The opportunity of articulating the conception of a real project with a strong theoretical background was unique in my academic experience, allowing practice and theory to stimulate mutually and power a more solid development of the project. As my goal was to idealize a house for a specific client in a specific place, the research swung from the exploration of the essence of the phenomenology of the spaces and their link to our mind, thus having studied authors like Gaston Bachelard [1] or Juhani Pallasmaa [2], to the concern to create a mass that relates strongly with the landscape and the surrounding buildings. Álvaro Siza and Alvar Aalto were very important references on this last approach. All this process was supported by the execution of models that allowed a progressive critical stand to the different solutions and a more mature development of the proposal. The results obtained are condensed in the specific form of the house (Fig. 1), in the way how spaces connect or disconnect, how old and new cohabit in the same place and how our body is induced to move through the different spaces. Among countless aspects, the opportunity of articulating a practical work with a strong theoretical support in one unique exercise made me realize that, in general, contemporary architecture privileges the sense of the sight and disregards the intimate relation between form and body. To establish more empathy between inhabitant and construction, it is crucial that our body is regarded as the central mass in the conception of all the habitable areas, assuming that the movement that we make between spaces is as important as the experience of sitting or lying down. The dynamism of moving gives us the real perception of the whole. In this way, senses play a central role because they allow us to feel the different intensities of the textures, forms or light.

Fig. 1 – Model of the site with the proposal

References: [1] Bachelard, Gaston (1998), A Poética do Espaço, Martins Fontes, São Paulo. [2] Pallasmaa, Juhani (2006), Los Ojos de la Piel, Editorial Gustavo Gili, Barcelona.

50

From RCR experience to a project of a small house in the mountain

F. Costa Leite 1, N. Brandão Costa 2

1 Faculty of Architecture, University of Porto, Portugal. 2 Faculty of Architecture, University of Porto, Portugal.

This work, which was developed concerning my Final Thesis for the Degree in Architecture of FAUP, consists in working up a house project for a farm keeper in the area of Gerês, north Portugal. As the thesis occurs after the experience got during the curricular training in RCR Arquitectes, it also includes a critical reflection on the works done by these architects, which are mostly located in the natural volcanic park of Garrotxa, north Catalonia. The reflection of the first chapter focus particularly on the work method of RCR, on the relation that their works establish with the landscape as a reference for their architecture, and on the materiality present on their whole work. To develop the project presented on the second chapter, an analysis of study cases was done, which enabled to understand the diversity of possible solutions in solving the program in question; a particular attention was also paid to the specific characteristics and problems of the site as being subjects and coordinates common to the trial of every solution. The project intends to solve an increasing need for a supporting structure in a farm on the banks of river Cávado that could function as a shelter for a gardener, a farmer, a friend or a relative. This way it came the opportunity of imagining a small retreat, suitable for a night and at the same time comfortable for an entire life (Fig. 1). The protect strategy starts from a study of the site, which precedes the design and which allowed to identify the main problems of the entire property, having had a great influence on the selection of the specific location, as well as on the solution principles. To achieve the final draw of the house for a farm keeper it was important the reflection on RCR Arquitectes’s work that reflects the lessons learned in that great laboratory, which was and still is, the geographical area where they have always been living. It was also important the organised and systematic research on architectural exercises with similar characteristics to that one defined in this project. The chosen examples contributed individually to the achievement of several hypotheses on the composition of the small house, as well as different ways of interacting with the surrounded areas. The project could then achieve its own mechanisms being the design the main tool and finding in the program and in the site, its most important motivations.

1. Project Model_scale 1:50

51

Casa Comum [common/collective/communitarian house/home] Architectures for an Intermediary Space

Nuno Travasso

Faculty of Architecture, University of Porto, Portugal.

This research pursuits an idea of collective dwelling. An idea which is not strictly related to a type or a form or an image, but which concerns a way of living and the essence of the space that provides lodging for it. Such as the main purpose of building a house is to create a home, also the design of collective housing should have as main purpose the materialization of an idea which is yet to be defined. “Casa Comum” (common home) is one approach to this urgently needed idea. This study is a domain with an identity of its own created by the constant interaction of multiple external fragments. These fragments are here called informants. They are the material elements that support this research and they can be divided in three types: bibliography, facts (specific spatial situations) and personal experience. In search of what is (or what could be) the role of collective dwelling in the construction of the city and in the definition of the way we inhabit the world, the research started with an analysis of the way man relates with the world and with the contemporary context. From this analysis it was realized that man inhabits only what he is capable of understand and control. It was also realized that each day it is harder for us to understand and control the world we live in as it is presented to us: fractured, built by nomadic and ephemeral fragments incapable of creating a coherent unity. A world that is the result of two opposed but intensely related movements: individualization and globalization. The result of these two movements is the gradual emptying of all intermediary scales: of all groups and communities, of all spaces in between. Hence the necessity of the establishment of an intermediary realm between the intimate and the global domains, as a way of creating the idea of a continuous, articulated and understandable world. If each man sees his home as the centre of his own world and if houses – and in specially, collective housing – are the material of which cities are mainly built, so collective dwelling can and should have a main role on the construction of this idea of world. “Casa Comum” is therefore presented as an expansion of the idea of home beyond the private realm. It proposes the creation of an inhabitable sphere intimately connected to the idea of home on the common domain. In other words, the establishment of a space and a community external but contiguous to the private space with which the inhabitant should identify himself and to which he should feel to belong. In this way it is possible to define the needed intermediary realm. The idea of “Casa Comum” is built on four main concepts – meeting space, expanded home, intermediary space and construction of the city – associated to four architectural principals – atmosphere, definition, articulation and prosthesis – based on which several design mechanisms are presented. In this way it is created a model which is, at the same time, construction of the idea of “Casa Comum” and means for the materialization of this idea. A model that establishes itself as an idea of architecture and as basis for a future architectural practice.

52

The Courtyard and Siza’s Houses

H. Henriques 1

1 Department of Architecture, Faculty of Architecture, University of Porto, Portugal.

The aim of this study was to attend the issue of the courtyard housing. The houses of Álvaro Siza serve as a motto for its deepening. From the analysis of these works it should be possible to recognize the importance that the author gives to the courtyard, as a mean for the settlement and link to the place. From an overview about the important role that the courtyard represents in the history of housing and of some of the recent research that has been over in the modern movement, we begin with an individual analysis of each house. The confrontation and comparison with other examples becomes necessary in order to understand the various means and ways of use of the courtyard in the domestic space. The six houses in this study are not in the same region, although a great part is placed in the north of Portugal. They appear along the architectural production of Álvaro Siza. Sometimes they reveal themselves as points of change or development in his language and method - an evolution carried through the Portuguese constructive tradition, the influences of some authors of the Modern Movement and the characteristics and cultural identity of each place. The variance of elements and materials is produced facing a closed composition, a sense of intimate environment of the house. The courtyard remains as an archetype that shapes the space. The study of the courtyard in the work of Siza is made with the desire to understand the patio as his way of thinking and doing architecture, looking for to portray, in the study cases, the features, expressions and feelings going through the project of interiorized houses. With the impossibility of a complete analysis of the courtyard throughout all his work, the chosen houses must be seen as examples that allow a particular approach to the issue. The study of those houses allows the observation of the formal and expressive mechanisms that separate or approach the house to what involves it. The analysis will allow, perhaps, to decipher the intentions and designs of Siza in choosing to turn the house on itself, sometimes to hide it from the outside look, perhaps searching for a general harmony of the space based on a relative or partial autonomy and an internal logic. The patio allows the understanding of the house as an area of protection and refuge from the outside world, but also as an area of representation - a static core where various activities take place but also as a mean to divide and create hierarchies and gradations between private and public, between the inside and outside. The research of houses of a single author led us to a reflection on the predominance of the courtyard in his work and, more particularly, about the continuity in his architecture. The unquestionable quality and beauty of the examples led us to an experienced learning. At the beginning of our work, the observation and visit of the houses showed the simplicity of the occupation of the site and the naturalness with which each space is opened to inside - a genuine synthesis of similar ways of life prevailed for the clear conception of the house as interior space.

53

The silence and the sky: seclusion and architecture. Four courtyard-houses

A. Silva Fernandes 1, C. Machado2

1 Lic. Architecture, MSc Student, Faculty of Architecture, University of Porto, Portugal. 2 Assistant Professor, PhD., Faculty of Architecture, University of Porto, Portugal.

The work being reported, based on the essay “The silence and the sky”[1], aimed to explore silence, in the context of architecture, as a void that completes the meaning of words and as space for thought and meditation. The courtyard in the house, as a medium for expressing the ability to establish a space for introspection and a connection between sky and earth, in a representation of the sacred and aspiration to perfection that is present throughout history, is used as a case-study. Through four themes – light, limit, matter and memory – the work focuses on the courtyard in cultural and historical references, using courtyards designed by Louis Kahn, Mies van der Rohe, Luís Barragán and Álvaro Siza, but mentioning many others, as a pretext for discussing architecture and poetry of space. The courtyard establishes a physical and symbolic unification of sky and earth. It is, at the same time, a recurrent and a versatile element, allowing adaptation to different conditions and generations, in different cultures and geographical locations. Evolving from an intuitive organization, calls upon primitive instincts, as the myth involved in the delimitation of the territory: creates a space for protection, introversion, of the imaginary. “It is so old that sensations inherited from man’s cave-dwelling days have been symbolized in it. It symbolizes femininity in the house and home; it is a spatial symbol of inwardness.”[2] As Heidegger explains, and as it is defended in the essay, poetry is connected with the human action of domination over the territory, of dwelling: “poetry opens the dwelling life of man”[3]. In a way, poetry of space is a kind of sacralization: even though not really connected with religion, gains a special meaning, even in a profane world, through memory or dream, forming something sacred, not necessarily divine, but human and personal. “Man, as man, has always measured himself with and against something heavenly.”[4] It is the acknowledgment of the unexplainable and the immeasurable, and the need for protection and seclusion, that is found in human nature, and thought to be reflected in architecture, namely in the courtyard. References: [1] Fernandes, Ana Silva (2007), O silêncio e o céu. Reclusão e Arquitectura. Quatro Casas-Pátio, Faculty of Architecture, University of Porto. This essay was written for the Award of the Degree in Architecture, University of Porto, and was supervised by the second author. [2] Blaser, W. (1985), Atrium – Lichthöfe Seit Fünf Jahrtausend – Five Thousand Years of Open Courtyards, Wepf & Co. AG, Verlag, Basel, p. 7. [3] Heidegger, M. (1971), Poetry, Language, Thought, Harper & Row, New York, Introduction, p.xii. [4] Heidegger, M. (1971), Poetry, Language, Thought, Harper & Row, New York, p.221.

54

PARALLEL ORAL SESSIONS II – POS.II.3 THURSDAY, FEB. 21

II.3 - NANO & SUPRAMOLECULAR CHEMISTRY Marcela Segundo & Adrián M.T. Silva 35 Effect of metals on the AtPreP1 peptidasome activity J. Pessoa, H.G. Bäckman, S. Bhushan, E. Glaser 38 Liposome-incorporated Zanthoxylum tingoassuiba essential oil: preparation and characterization C. Detoni, B. Sarmento, D. Ferreira, E.C.M. Cabral-Albuquerque 90 Anchoring of macrocycle compounds onto nanostructured carbon materials for catalytic applications Dalila Teixeira, Delfina Barros, Susana L. H. Rebelo, M. F. R. Pereira, J. J. M. Órfão, Cristina Freire 132 Characterizing the intermetallic formed during ball attach process Paulo Pereira, Rúben Santos, Maria M. Barbosa, Cátia Almeida 134 Functional Insulin Quantification upon its Nanoencapsulation M. J. Barbosa, J. Faria, O. Queirós, A. Ribeiro, R. Moreira Temperature and composition effects on the micelle formation by a catanionic surfactant: a surface 142 tension and conductivity study R. F. Fernandes, B. F. B. Silva, E. F. Marques

Effect of metals on the AtPreP1 peptidasome activity

J. Pessoa1,2, H.G. Bäckman2, S. Bhushan2 and E. Glaser2

1 Institute for Molecular and Cell Biology, Faculty of Sciences, University of Porto, Portugal. 2 Department of Biochemistry and Biophysics, Faculty of Science, University of Stockholm, Sweden.

In eukaryotic cells, proteins are targeted to their correct organelle by targeting peptides, which are amino acid extensions located in the N-terminal portion of the precursor protein. After the protein has reached its final intracellular destination, targeting peptides are cleaved off, and because of their cell toxicity, they must be removed. A novel metalloprotease that efficiently degrades targeting peptides in mitochondria and chloroplasts was identified by our group, and named Presequence Protease, PreP [1]. In Arabidopsis thaliana there are two isoforms of PreP (AtPreP1 and AtPreP2) that show an 86% homology at the amino acid sequence level [2]. AtPreP1 is composed of 995 amino acids, and contains a zinc-binding motif in its catalytic site. It also contains two magnesium-binding sites, one located on the inner side of the peptide binding cavity and another on the outer surface of the enzyme [3]. Magnesium (or calcium) has been shown to be needed for catalysis. The aim of this project is to understand the role of magnesium for the degradation activity. For that, the activity of AtPreP1 variants (which are unable to bind magnesium) was measured and compared to the wild-type. Three AtPreP1 variants, one in which both magnesium-binding sites were changed, and two in which only one magnesium-binding site was altered, were generated, overexpressed in bacteria and purified. Their activity was tested using different concentrations of magnesium, calcium or zinc and different substrates. The optimal metal concentrations for catalysis were estimated. Both the single and double AtPreP1 variants were inactive against a 54 amino acid residue peptide corresponding to the ATPase F1β presequence. However, when the degradation was tested with five other shorter peptides, the results showed that the activity was substrate-specific and that it may depend on the substrate length.

References: [1] Ståhl, A., Moberg, P., Ytterberg, J., Panfilov, O., Brockenhuus von Löwenhielm, H., Nilsson, F. and Glaser, E. (2002), Isolation and Identification of a Novel Mitochondrial Metalloprotease (PreP) that Degrades Targeting Presequences, J. Biol. Chem., 277 (44), 41931-41939. [2] Ståhl, A., Nilsson, S., Lundberg, P., Bhushan, B., Biverståhl, H., Moberg, P., Morisset, M., Vener, A., Mäler, L., Langel, U., and Glaser, E. (2005), Two Novel Targeting Peptide Degrading Proteases, PrePs, in Mitochondria and Chloroplasts, so Similar and Still Different, J. Mol. Biol., 349, 847–860. [3] Johnson, K.A., Bhushan, S., Ståhl, A., Hallberg, B.M., Frohn, A., Glaser, E. and Eneqvist, T. (2006), The closed structure of presequence protease PreP forms a unique 10 000 Å3 chamber for proteolysis, EMBO J., 25 (9), 1977-1986.

56

Liposome-incorporated Zanthoxylum tingoassuiba essential oil: preparation and characterization

C. Detoni 1,2, B. Sarmento2, D. Ferreira2 and E.C.M. Cabral-Albuquerque1

1Medical Subject Research Laboratory, Pharmacy Faculty, Federal University of Bahia, Brasil. 2 Department of Technology, Faculty of Pharmacy, University of Porto, Portugal.

Alfa-bisabolol and essential oils, such as Chamomilla recutita and Zanthoxylum tingoassuiba oils, are of pharmaceutical and cosmetic industries interest for its known antimicrobial and antiphlogistic actions[1,3]. Recent investigations also found a possible anticancer activity of alfa-bisabolol on glioma cells[1,2]. Despite the known activities of Zanthoxylum tingoassuiba essential oil as a promising non-toxic drug, it has presented low biological in vitro activity because of its low solubility in water. One of the most recent strategies for overcoming this drawback is a reversible association of the essential oil to liposomes[3,4]. The objective of this research work was to prepare and characterize Zanthoxylum tingoassuiba essential oil-loaded liposomes regarding further evaluation of liposomal inclusion on the antimicrobial activity of this essence. The essential oil was incorporated in multilamellar (MLV) and unilamellar (LUV) vesicles of synthetic dipalmitoil-phosphatidylcholine (DPPC) by the thin-lipid film hydration and high pressure homogenization-extrusion methods respectively. The formulations were characterized regarding the mean particle size by photon correlation spectroscopy, surface charge by laser Doppler Anemometry, the morphology by transmission electron microscopy, alfa-bisabolol incorporation efficiency and release by high-performance liquid chromatography (HPLC) with UV detection. The results obtained show that the produced liposomes have spherical shape. The mean size of MLV´s was 10.6 μm and of LUV´s was 0.26 μm. The liposomes electric charge did not change after oil incorporation, indicating that the oil was not adsorbed on the surface. The incorporation efficiency varied with the preparation technique, being over 50% for MLV but lower for LUV. The in vitro release profile presented a sustained and incomplete release of MLV-incorporated EO for 24h, suggesting that EO-loaded liposomes will be useful in pharmaceutical application to enhance essential oil targeting to cells that can not cross the cytoplasmatic barrier. Zanthoxylum tingoassuiba essential oil was successfully incorporated in liposomes and the results confirm the potentiality of MLV formulations for pharmaceutical and cosmetic use. References: [1] Perbelin, L., Gottardo,R., Caprini, A., Bortolotti,F., Marioto, S. and Tagliaro, F. (2004), Determination of alpha-bisabolol in human blood by micro-HPLC–ion trap MS and head space- GC–MS methods, Journal of Chromatography, 812 , 373–377. [2] Cavalieri, E., Marioto, S.,Fabrizi, C.,Prati, A.C.,Gottardo, R.,Leone, S., Berra, L.V.,Lauro, G. M., Ciampa, A.R. and Suzuki H. (2004), α-Bisabolol, a nontoxic natural compound, strongly induces apoptosis in glioma cells, Biochemical and Biophysical Research Communications, 315 (3), 589-594. [3]Cabral-Albuquerque, E.C., Hohlemweger, S., Detoni, C., Matos, J., Sampaio,C., Barros, T., Veloso, E.(2007) Essential Oil from Zantoxyllum tingoassuiba Loaded into Multilamellar Liposomes Useful as Antibiotic and Antimicrobial. In: 34th Controlled Release Annual Meeting & Exposition,Long Beach/CA. CD-Rom transactions of the 34th Annual Meeting & Exposition - Controlled Release Society.

57

Anchoring of macrocycle compounds onto nanostructured carbon materials for catalytic applications

Dalila Teixeira,1 Delfina Barros,1 Susana L. H. Rebelo,1 M. F. R. Pereira,2 J. J. M. Órfão,2 Cristina Freire1

REQUIMTE, Departamento de Química, Faculdade de Ciências, Universidade do Porto, 4169-007 Porto.

The immobilisation of transition metal complexes with catalytic properties onto supports is a theme of intense research, due to its importance towards the goals of Green Chemistry. Porphyrins and salen-type metal complexes have been succeeded on homogeneous catalytic oxidations. However, their catalytic properties (catalyst stability and recycling) can be further improved by immobilisation onto solid supports. Carbon materials are unique supports as they can provide a variety of oxygen surface groups at the edges/defects of graphene sheets that can be tailored by adequate thermal/chemical treatments, besides the inherent chemical/physical reactivity associated with the graphene sheets which are hydrophobic and present low polarity and a rich -electron density. In this project we endeavour to synthesise and immobilise porphyrins and salen-type metal complexes, Figure 1, that exhibit catalytic properties in the oxidation of organic pollutants, onto nanostructured carbon materials: multi-walled carbon nanotubes (MWCNTs), carbon xerogels (CX) and templated carbons (CMK).

Ar Cl a) b) Ar = N N NN Cl Mn Ar M Ar O O N L N F F Cl HO OH Ar = F Ar F F M = 2H, Zn(II), Ni(II), Cu(II) Au(III), Fe(III), Mn(III) Figure 1: Molecular structures of a) porphyrins and b) salen-type metal complexes.

In this work we report the preparation and the characterisation of several porphyrins and salen-type ligands and the corresponding metal complexes, M=Fe(III), Mn(III), Au(III), Ni(II), Cu(II) and Zn(II). These compounds will be grafted to the untreated carbon materials (MWCNT, CX and CMKs) and fullerenes (reference material) by covalent bonding. The covalent anchored procedure will be achieved by utilizing 1,3-dipolar addition of azomethine ylides and cyclopropanation under Bingel reaction conditions.

Acknowledgments: This work is funded by Proj. IPG58 de Investigação Científica na Pré- Graduação 2007 (Universidade do Porto e Caixa Geral de Depósitos).

58

Characterizing the intermetallic formed during ball attach process

Paulo Pereira a,*, Rúben Santos a, Maria M. Barbosa a, Cátia Almeida a

a** Dep. de Engenharia Metalúrgica e de Materiais, Faculdade de Engenharia, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal * [email protected]

Continuously miniaturization of electronic components requires the most advanced technologies in which electrical response is a crucial factor to success. In this study we characterize the interface of an attachment between the copper conductors of a memory and a solder ball which allows further bonding to printed circuit modules, see figure 1a). , This connection, made of diferent layers, has a capital importance to ensure a good electrical and mechanical performance. The attachment uses a 10m thick intermediate layer of nickel deposited over a copper pad and protected by a thin gold film. The solder, with a spherical shape and composition of Sn-1.0Ag-0.5Cu, is placed on the gold film and the assemblage is heated up to the brazing temperature. At the interfacial zone, between the nickel and the solder ball, a continuous layer of an intermetallic compound of Sn-Ni-Cu is formed. This intermetallic layer has 1 to 2 m thickness as seen on figure 1b). Some particles of this compound are also observed at inner zones of the solder ball (see arrow on figure 1b), indicating a preferential copper and nickel diffusion path inside the ball. By an AFM analysis it was possible to perform nanoindentation tests, shown on picture 1c). The indentation size variation between Sn braze alloy and intermetallic layer clearly states the hardness increase at the interface.

Figure 1. a) SEM image of the ball attach bonding; b) SEM image of interface showing the intermetallic compound formed between the Ni (darker layer) and the Sn braze alloy (lighter area); c) AFM image of the indentations in the braze alloy and intermetallic compound.

Acknowledgement The materials characterized in this work were kindly supplied by Qimonda S.A. ** The authors are undergraduate students of the 4th year of Master in Metallurgical and Materials Engineering.

59

Functional Insulin Quantification upon its Nanoencapsulation

M. J. Barbosa1,3*, J. Faria2,3*, O. Queirós3, A. Ribeiro3 and R. Moreira3

1 Faculty of Science, University of Oporto and ICBAS, University of Oporto, Portugal. 2 Faculty of Science, University of Oporto, Portugal. 3 Instituto Superior de Ciências da Saúde – Norte, Centro de Investigação em Ciências da Saúde (CICS), Grupo de Biologia Molecular e Celular, CESPU, Portugal. * These authors contributed equally to this work.

Insulin is the most import drug used in the therapeutics of diabetes mellitus, a metabolic disorder with increasing prevalence. Several alternative routes of administration, other than the subcutaneous one, are currently under development, in order to improve patients’ quality of life [1]. Among them, insulin encapsulation within biodegradable polymer particles regarding its oral administration constitutes a promising strategy [2]. In the present study, an in vitro methodology was developed to evaluate the preservation of the hormone’s functionality following its submission to nanoencapsulation. The methodology was optimized using commercially available insulin, through rat L6 myoblasts stimulation with different hormone concentrations and for distinct periods of time. Detection of Akt phosphorylated form through Western blotting assays was used as an indicator of an effective stimulation by insulin and, thus, of the retention of its active conformation [3]. Akt/PKB, one of insulin signalling pathway phosphorylation products, is a protein kinase involved in many of the hormone’s biological actions, including glucose transport and modulation of gene expression. Assays using insulin recovered from nanoparticles revealed that the emulsification/internal gelation technique preserves part of insulin’s functionality. The methodology may be of use as a rapid, ethical, specific and conclusive means of bioactivity screening in pharmaceutical formulations containing the hormone.

References: [1] Gerich, J.E. (2002), Novel insulins: expanding options in diabetes management, The American Journal of Medicine, 113, 308-316.

[2] Silva, C.M., Ribeiro, A.J., Figueiredo, I.V., Gonçalves, A.R. and Veiga, F. (2006), Alginate microspheres prepared by internal gelation: development and effect on insulin stability, International Journal of Pharmaceutics, 311, 1-10.

[3] Patel, N., Craddock, B.L., Staniforth, J.N., Tobyn, M.J. and Welham, M.J. (2001), Spray-dried insulin particles retain biological activity in rapid in-vitro assay, Journal of Pharmacy and Pharmacology, 53, 1415-1418.

60

Temperature and composition effects on the micelle formation by a catanionic surfactant: a surface tension and conductivity study

R. F. Fernandes, B. F. B. Silva and E. F. Marques

Centro de Investigação em Química, Department of Chemistry, University of Porto, Portugal.

Surfactants are molecules with a polar headgroup, which favors contact with water (hydrophilic part), and an apolar tail, which avoids contact with the solvent (hydrophobic part). As a consequence, they have two important properties: they adsorb at polar/apolar interfaces, reducing the interfacial tension; they self-assemble in water into different kinds of aggregates, such as micelles and liquid crystals. The formation of micelles is a spontaneous and cooperative process that occurs at a given concentration, the critical micelle concentration (CMC), for constant temperature. For an ionic surfactant, the CMC is influenced by factors such as temperature, ionic strength and pH. For anionic/cationic surfactant mixtures, the mixing ratio is also a determinant parameter. Typically, there is a significant decrease of the mixture CMC relative to the CMC of the individual surfactants [1], a relevant property for practical applications (e.g. in detergent formulation). In this work, we have used surface tension and electrical conductivity techniques in order to investigate the micellization behavior of a newly synthesized surfactant, hexadecyltrimethylammonium octylsulfonate (TASo). This surfactant belongs to the class of catanionics and is obtained by the equimolar pairing of a long chain cationic molecule and a short chain anionic one, followed by removal of the remaining counterions (sodium bromide) [2]. TASo has several unusual and interesting properties, such as being water- soluble at room temperature, to yield micelles as the first aggregate, followed by spontaneous liposomes [3], a type of colloidal aggregates extremely useful in biomedical applications, such as drug delivery and gene therapy. We have investigated the effect of temperature, salt and composition (addition of excess ionic surfactant) on the CMC and the degree of counterion dissociation of the micelles of TASo. This allows the determination of the enthalpy, entropy and Gibbs function of micellization. A comparative study of the ionic surfactant sodium octylsulfonate was also carried out. It is observed that the CMC of TASo increases with temperature in a similar way to non-ionic surfactants. However, in all other properties the surfactant behaves like an ionic surfactant, e.g. the CMC decreases significantly with salt addition. This is rationalized by the fact that TASo acts as a weakly dissociated molecule [3], due to the much higher solubility of the shorter chain anionic component, leaving the micelles with a residual electrostatic charge. References: [1] Khan, A., Marques, E. F. (1997), Catanionic Surfactants, in Specialist Surfactants, Blackie Academic and Professional, London. [2] Silva, B.F.B. and Marques, E.F. (2005) Thermotropic behavior of an asymmetric chain length catanionic surfactant, J. Colloid Interface Sci., 290, 267-274. [3] Silva, B.F.B, Marques, E.F. and Olsson U. (2007) Lamellar Miscibility Gap in a Binary Catanionic Surfactant-Water System, J Phys. Chem B, 111, 13520-13526.

61

PARALLEL ORAL SESSIONS III – POS.III.1 THURSDAY, FEB. 21

III.1 - LIFE & HEALTH II Maria José Oliveira & Luís Miguel Madeira 26 Phenolic compounds, organic acids and antioxidant properties of Rumex induratus L. Guerra, C. Pereira, R. Gonçalves, P. B. Andrade, R. M. Seabra, P. Valentão MUC1 overexpression is associated with distant metastases development in canine mammary 37 carcinomas de Oliveira JT, Pinho S, Matos AJ, Lopes C, Barros R, Hespanhol V, Reis C, Gärtner F 42 Analysis of the efficiency of the mitotic checkpoints in glioblastoma cell lines A. Nascimento, E. Logarinho, O. Martins, R. M. Reis, H. Bousbaa Prenylated and bromoalkylated xanthones as potential antitumor agents: synthesis and biological 104 activities A. Paiva, E. Sousa, M. Pinto, A. Camões, N. Nazareth, M. S. J. Nascimento 157 P-cadherin: role in breast cancer cell migration and invasion Ribeiro AS, Paredes J, Correia AL, Schmitt F Impairment of retrograde signalling via adenosine in toxin-induced Myasthenia gravis: cross-talk 162 with muscarinic autoreceptors Diogo Trigo, Ana Sá-e-Sousa, Tiago Morais, Maria Alexandrina Timóteo, Teresa Magalhães-

Cardoso, Laura Oliveira, Paulo Correia-de-Sá

Phenolic compounds, organic acids and antioxidant properties of Rumex induratus

L. Guerra1, C. Pereira1, R. Gonçalves1, P. B. Andrade1, R. M. Seabra1, P. Valentão1

1 Department of Pharmacognosy, Faculty of Pharmacy, University of Oporto, R. Aníbal Cunha, 164, 4050-047 Oporto, Portugal.

Rumex induratus Boiss. & Reuter is an endemic Iberian herb, which fundamentally develops in the thermo Mediterranean region. It grows spontaneously in Northeast Portugal where its leaves are highly consumed. This species is very appreciated in salads and, to attenuate its acidity, is dressed with olive oil and sometimes mixed with boiled potatoes. The chemical composition of aqueous extract of R. induratus leaves, in terms of phenolic compounds and organic acids, and its antioxidant activity against DPPH radical and a Reactive Nitrogen Species, nitric oxide ( NO), were studied.

Fig. 1. HPLC-DAD phenolic profile of R. iduratus aqueous extract. (1) caffeoyl-hexoside; (2) p-coumaroyl- hexoside; (3) feruloyl-hexoside; (4) sinapoyl-hexoside; (5) 6-C-hexosyl- quercetin; (6) 8-C-hexosyl-luteolin; (7) 6-C-hexosyl-luteolin; (8) 6-C- hexosyl-apigenin; (9) 3-O-hexosyl- quercetin; (10) 3-O-rutinosyl- quercetin; (11) 7-O-hexosyl- diosmetin; (12) 3-O-rutinosyl- isorhamnetin; (a) and (b) unidentified O-glycosil-C-glycosilflavones.

Minutes

Twelve phenolic compounds were identified by HPLC/DAD, based on previous work [1] (Fig. 1). Six organic acids were determined by HPLC-UV: oxalic, citric, malic, ascorbic and shikimic acids. Some variations in the phenolics and organic acids profiled are discussed, regarding the influence of the origin, developmental stage and crop season of the samples. R. induratus aqueous extract revealed scavenging activity against both radicals, in a concentration dependent manner. 7-O-Glucosyl luteolin, a structurally related compound and oxalic acid were also tested. The phenolic compound displayed a strong activity while oxalic acid had no effect. Thus, the antioxidant capacity found for R. induratus extract can be mainly ascribed to this class of compounds. References: [1] Ferreres, F., Ribeiro, V., Izquierdo, A.G., Rodrigues, M.A., Seabra, R.M., Andrade, P.B., Valentão, P., (2006) Rumex induratus Leaves: an Interesting Dietary Source of Potential Bioactive Compounds. J Agric Food Chem, 54, 5782-5789.

63

MUC1 overexpression is associated with distant metastases development in canine mammary carcinomas

de Oliveira JT 1,2, Pinho S 1,2, Matos AJ2, Lopes C2, Barros R1,3, Hespanhol V3, Reis C1,3 and Gärtner F1,2

1Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Rua Dr Roberto Frias s/n, 4200-465 Porto, Portugal 2Institute of Biomedical Sciences of Abel Salazar (ICBAS), University of Porto, Largo Prof. Abel Salazar, 2, 4099-003 Porto, Portugal 3Medical Faculty, University of Porto, Alameda Prof. Hernâni Monteiro 4200-319 Porto, Portugal

Canine mammary tumours affect mainly older bitches and comprise approximately 25- 50% of all their tumours, 40-50% being malignant [1,2]. MUC1 is overexpressed in human breast cancer and contributes to carcinoma progression [3,4]. High MUC1 expression is linked to a poorer prognosis [5]. MUC1 expression had never been described in canine mammary tumours (CMT) before. The aims of this work where: to characterize MUC1 expression in CMT and to evaluate its relationship with clinicopathological features such as tumour histological type, mode of growth, tumour grading, lymph node metastases and distant metastases. Fifty paraffin tumour sections were examined for MUC1 immunostaining patterns. Immunohistochemistry technique, using antibody C-20, determined MUC1 expression. Associations between clinicopathological features and MUC1 expression were analysed. All tumours showed MUC1 immunostaining. In normal adjacent mammary gland tissue, MUC1 was detected in the apical cell membrane. In the carcinomas MUC1 was detected in the cytoplasm (52.0%), circumferential membrane (2.0%), or a mixture of both patterns (46.0%). The follow up period was of 2 years, during which 10 distant metastases were confirmed. We observed that a total of 20.4% of tumours gave rise to distant metastases, among these and most importantly 100% showed significantly high (≥50% positive cells) MUC1 expression (p= 0.03). In the bitch, high MUC1 expression was significantly associated to higher metastatic development. Our findings indicate, for the first time in CMT, that MUC1 may be an important prognostic marker in these tumours as it is in human breast cancer. References: [1] Sorenmo, K. (2003), Canine mammary gland tumors, Vet Clin Small Anim, 33, 573-596. [2] Owen, L.N. (1979), A comparative study of canine and human breast cancer, Invest Cell Pathol, 2, 257-275. [3] Baldus, S.E., Engelmann, K., Hanisch, F.G. (2004) MUC1 and the MUCs: a family of human mucins with impact in cancer biology, Crit Rev Clin Lab Sci, 41, 189-231. [4] Croce, M.V., Isla-Larrain, M.T., Rua, C.E., Rabassa, M.E., Gendler, S.J. (2003), Patterns of MUC1 tissue expression defined by an anti-MUC1 cytoplasmic tail monoclonal antibody in breast cancer, J Histochem Cytochem, 51, 781-788. [5] Rakha, E.A., Boyce, R.W., El-Rehim, D.A., Kurien, T., Green, A.R., Paish, E.C. et al. (2005), Expression of mucins (MUC1, MUC2, MUC3, MUC4, MUC5AC and MUC6) and their prognostic significance in human breast cancer, Mod Pathol, 18, 1295-1304.

64

Analysis of the efficiency of the mitotic checkpoints in glioblastoma cell lines

A. Nascimento1,3, E. Logarinho2, O. Martins2, R. M. Reis2 and H. Bousbaa3

1 Faculty of Science, University of Oporto and ICBAS, University of Oporto 2 Life and Health Sciences Research Institute, School of Health Sciences, University of Minho 3 Instituto Superior de Ciências da Saúde – Norte, Centro de Investigação em Ciências da Saúde (CICS), Grupo de Biologia Molecular e Celular, CESPU

The mitotic checkpoint modulates the timing of anaphase initiation in response to improper alignment of chromosomes at the metaphase plate. The BUB and MAD gene families encode proteins which are part of a large multi-protein complex which are believed to be key components of the checkpoint regulatory pathway [1][1]. Failure of this surveillance system can lead to genomic instability and could be responsible for the increased incidence of aneuploidy and possible driving forces in, tumorigenesis [3]. Glioblastomas are the most common and malignant form of primary adult brain tumors[4]. These tumors are characterized by marked chromosomal instability, with gains and/or losses of chromosomes, referred as aneuploidy [6]. In order to get insight to the mechanism that lead to aneuploidy in glioblastomas, we assessed the efficiency of the mitotic checkpoints and possible molecular alterations in two glioblastoma cell lines. To evaluate mitotic arrest efficiency, mitotic index (percentage of viable cells arrested in mitosis) was determined, in untreated as well as in cell cultures treated with the microtubule-disrupting drug nocodazole for 16 hours, by cell-rounding under phase contrast microscopy. The molecular alterations in checkpoint genes were evaluated at the protein levels by Western blotting analysis. For that, total protein extracts were prepared from glioblastoma cells and the protein amounts were determined by quantitative immunoblotting using specific antisera. The protein expression levels were compared to those in HeLa cells, using the alpha-tubulin levels as reference standards. Concerning the efficiency of the mitotic checkpoint, glioblastomas showed a low mitotic index, a lack of mitotic arrest when incubated with nocodazole together with an alteration in the expression levels of the mitotic checkpoint proteins examined. These results confirm our suspicion of a deficient mitotic checkpoint in glioblastoma cells which, through premature onset of anaphase, lead to chromosome mis-segregation. Our finding might provide an explanation for the chromosomal defects seen in glioblastoma. [1] May, K. M. e Hardwick, K. G. (2006). The spindle checkpoint in Journal of Cell Science 119(Pt 20), 4139-42. [2] McGowan, C. H. (2003). Regulation of the eukaryotic cell cycle in Progress in Cell Cycle Research 5: 1-4. [3] Cahill, D. P., et al. (1998).Mutations of mitotic checkpoint genes in human cancers in Nature, 392(6673), 300-3. [4] Isaka, T., et al. (2003). Chromosomal variations within aneuploid cancer lines in Journal of Histochemistry and Cytochemistry, 51(10): 1343-53. [5] Kleihues P, and Cavenee WK; The WHO classification of tumors of the nervous system, International Agency for Research on Cancer, 2000. [6] Draviam, V.M. et al., (2004) Chromosome segregation and genomic stability in Current Opinion in Genetics and Development 2004 Apr;14(2), 20-5

65

Prenylated and bromoalkylated xanthones as potential antitumor agents: synthesis and biological activities

A. Paiva1,3, E. Sousa1,3, M. Pinto1,3, A. Camões2,3, N. Nazareth2,3 and M. S. J. Nascimento2,3

1 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 Department of Microbiology, Faculty of Pharmacy, University of Porto, Portugal. 3 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal.

In order to improve the growth inhibitory effect of 3,4-dihydroxyxanthone (1) on human tumor cell lines [1], the synthesis of new prenylated and bromoalkylated derivatives was planned. Three prenylxanthones (2-4), as well as two bromohexyloxyxanthones, (5, 6) were obtained by different synthetic pathways (Fig. 1). The structures of compounds 2-6 were established by spectroscopic methods (IR, NMR of 1H and 13C, HMBC and HSQC) and mass spectrometry.

O

BrCH2CHC(CH3)2

K CO ,DMF,reflux 2 3 O O OH 2 O O O

Br CH 2CHC( CH 3)2 O OH + K2CO3 ,DMF,r.t. O O OH O O 1 O OH 3 4

Br(CH2)6Br O O (drop-to-drop) + K CO ,DMF,r.t. 2 3 Br Br O O O O O OH Br 5 6 Fig. (1). Synthesis of prenylated and bromoalkylated xanthones.

The effects of xanthones 2-6 on the in vitro growth of estrogen receptor positive (ER +) MCF-7 and estrogen receptor negative (ER −) MDA-MB-231 cells were investigated. Prenylated compounds 2-4 and monobromoalkylxanthone 6 were more potent than the parent compound 1. Moreover, compounds 2-4 revealed selectivity against ER(+) MCF-7 cells. Compound 2, the most potent derivative (GI50 = 5 µM, MCF-7), showed an antiproliferative effect either on complete and steroid-free RPMI medium suggesting not to be dependent on estrogenic stimulation. The growth inhibitory action of the antiestrogen 4- hydroxytamoxifen in ER(+) MCF-7 cell line was strongly enhanced by compound 2. This is the first report of interactions between xanthones and an antiestrogen. References: [1] Pedro M., Cerqueira, F., Sousa, M.E., Nacimento, M.S.J. and Pinto, M. (2002), Xanthones as inhibitors of growth of human cancer cell lines and their effects on the proliferation of human lymphocytes in vitro, Bioorganic Medicinal Chemistry, 10 (12), 3725-3730. Acknowledgments: FCT (I&D, nº226/94), FEDER, POCI for financial support.

66

P-cadherin: role in breast cancer cell migration and invasion

Ribeiro AS1, Paredes J1,2, Correia AL2, Schmitt F 1,2

1 IPATIMUP, Institute of pathology and molecular immunology from University of Porto; 2 School for Health and Sciences, Minho University

Cadherins are calcium-dependent glycoproteins that mediate cell-cell adhesion, presenting a characteristic pattern of distribution in human tissues. In mammary gland, P-cadherin is only expressed in myoepithelial cells; however, its expression has been reported in breast carcinomas, where it is associated with high-grade histological tumours and with poor patient survival [1-3]. In order to elucidate the role of P-cadherin in breast cancer progression, we aimed to clarify the capabilities acquired by breast cancer cells, such as motility, migration and invasion, using as a model a breast cancer cell line (MCF-7/AZ) which was retrovirally transduced with human P-cadherin (MCF-7/AZ.Pcad). Using a wound healing assay, we demonstrated that P-cadherin overexpression was able to promote breast cancer cell migration, which was suppressed by a blocking antibody against this protein. Also cell motility, measured by time-lapse microscopy, was increased, and it was strongly associated with the formation of actin filopodia and lamellipodia, whereas its siRNA-specific silencing reduced the presence of these membrane protrusions. Besides our group has shown previously, that P-cadherin overexpression in breast cancer cells promotes in vitro cell invasion [4], in the present study we proved that this effect is P- cadherin specific, since invasion was decreased to basal levels upon the inhibition of this molecule. We then considered that this effect could be due to proteases secretion to the medium, and in fact MMP-1 and MMP-2 activity and expression showed to be increased in MCF-7/AZ.Pcad cells, as well as a soluble P-cadherin fragment. Furthermore, when parental cells where treated with the conditioned medium recovered from P-cadherin overexpressing cells, cell invasion was promoted, implying that MCF-7/AZ.Pcad cells secrete proteins to the medium that should be involved and facilitate cell invasion. Taken together, this work presents, a possible cellular mechanism explaining why P- cadherin expression is associated with a more aggressive phenotype and a poor patient survival in breast carcinomas.

References [1] Paredes J, Milanezi F, Viegas L, Amendoeira I, Schmitt FC (2002), P-cadherin expression is associated with high-grade ductal carcinoma in situ of the breast, Virchows Arch 440: 16-21, 2002 [2] Paredes J, Milanezi F, Viegas L, Amendoeira I, Schmitt FC (2002), P-cadherin expression is associated with high-grade ductal carcinoma in situ of the breast, Virchows Arch 440: 16-21, 2002 [3] Paredes J, Albergaria A, Oliveira JT, Jerónimo C, Milanezi F, Schmitt FC (2005), P- cadherin overexpression is an indicator of clinical outcome in invasive breast carcinomas and is associated with CDH3 promoter hypomethylation, Clin Cancer Res 11: 5869-5877, 2005 [4] Paredes J, Stove C, Stove V, Milanezi F, Van Marck V, Derycke L, Mareel M, Bracke M, Schmitt F (2004), P-cadherin is upregulated by ICI 182,780 in breast cancer cells and promotes invasion via its juxtamembrane domain, Cancer Res 64: 8309-8317, 2004.

67

Impairment of retrograde signalling via adenosine in toxin- induced Myasthenia gravis: cross-talk with muscarinic autoreceptors

Diogo Trigo1, Ana Sá-e-Sousa1, Tiago Morais1, Maria Alexandrina Timóteo1, Teresa Magalhães-Cardoso1, Laura Oliveira1 & PauloCorreia-de-Sá1

1Laboratório de Farmacologia e Neurobiologia, Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas de Abel Salazar – Universidade do Porto (ICBAS-UP), Portugal.

While adenosine acts predominantly as an inhibitory signal (via A1 receptors) under resting conditions, amplification of neuromuscular transmission depends on facilitation of acetylcholine (ACh) release via muscarinic M1 autoreceptors [1,2]. Upon increasing the stimulation frequency, predominant activation of adenosine A2A receptors counteracts the M1 positive feedback mechanism causing a shift on muscarinic neuromodulation towards the activation of inhibitory muscarinic M2 autoreceptors [2]. These complex receptor-receptor interactions, involving distinct second messengers and effectors [3], may offer the potential for regulating neurotransmitter exocytosis and, hence, minimization of transmission deficits observed in myasthenic patients. This prompted us to evaluate the role of adenosine and muscarinic autoreceptors in a rat model of toxin-induced Myasthenia gravis (TIMG). Wistar rats (70-100 g) were injected once every 48h with saline (controls) or α-bungarotoxin (α-BTX, an irreversible muscle-type α1 nicotinic receptor antagonist) (TIMG-model) for a period up to 6 weeks [4]. Dosage of α-BTX was adjusted by monitoring myasthenic symptoms. The procedures for measuring evoked [3H]-ACh release and diaphragm contractile responses were described previously. [3H]-ACh release was evoked by phrenic nerve stimulation with either 5 Hz-trains (750 pulses) or 50 Hz-bursts (5 bursts of 150 pulses, 20-s interburst interval). Fatigue tests were carried out using high frequency (50 Hz) intermittent (17 pulses per sec, during 3 minutes) nerve stimulation. In contrast with that observed in control animals, inactivation of endogenous adenosine with adenosine deaminase (ADA, 2.5 U/ml) and blockade of A1 receptors with 1,3-dipropyl-8-cyclopentyl xanthine (DPCPX, 10 nM) failed to increase [3H]-ACh release evoked by 5 Hz-trains. ADA (2.5 U/ml) decreased (- 18±6%, n=6) evoked [3H]-ACh release when the phrenic nerve of TIMG animals was stimulated with 50 Hz- bursts, but this effect was significantly (P<0.05) smaller than that observed in control rats (-54±8%, n=5). Likewise, the inhibitory effect of the selective adenosine A2A receptor antagonist, ZM 241385 (50 nM), was smaller in TIMG animals (-19±9%, n=4) as compared with controls (-46±7%, n=6). ZM 241385 (50 nM) enhanced fatigue of diaphragm contractions in control animals, but it was virtually devoid of effect in TIMG animals. No differences were found on the release inhibitory effect of pirenzepine (10 nM, an M1 receptor antagonist) in both control and TIMG rats. The muscarinic M2-mediated inhibition, revealed by facilitation of [3H]-ACh release evoked by 50 Hz-bursts in the presence of AF-DX 116 (10 µM), was significantly attenuated in TIMG animals (7±5%, n=5) as compared with controls (53±16%, n=5). Muscle paralysis due to selective blockade of muscle action potentials, with μ-conotoxin GIIIB (0.6 µM), suppressed stimulation- induced adenosine release measured by HPLC analysis. Data suggest that impairment of contractile activity in myasthenic rats reduces retrograde signalling mediated by adenosine outflow from the muscle, which causes profound changes in the control of neuromuscular transmission. These changes are particularly evident during high frequency nerve stimulation (50 Hz-bursts) and result from minimization of adenosine control of muscarinic autoreceptors favouring M1 facilitation of transmitter release while decreasing the M2 inhibitory tonus. [1] Correia-de-Sá et al., 1996, J. Neurophysiol., 79, 3910-19; [2] Oliveira et al., 2002, Eur. J. Neurosci., 15, 1728-36; [3] Oliveira & Correia-de-Sá, 2005, NeuroSignals, 14, 262-72; [4] Plomp et al., 1992, J. Physiol., 458, 487-99. This work was supported by FCT (FEDER funding, POCTI/CVT/43368/2001).

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PARALLEL ORAL SESSIONS III – POS.III.2 THURSDAY, FEB. 21

III.2 - ENVIRONMENT & TECHNICAL ASPECTS Lígia Afonso & Sofia Rodrigues 149 Contemporary condition: 'new' relationships between architecture and city Mariana Alves 126 Form and matter in the work of Rafael Moneo Rui Gonçalves 172 Architecture and climate. Site geography: Luanda and the work of Vasco Vieira da Costa M. Quintã, J. Quintão Crises e Reformas: o colonialismo ilustrado na Capitania de São Paulo e os esforços de 222 modernização do Governo Morgado de Mateus (1765-1775) Joana Salém Vasconcelos, Vera Lúcia Amaal Ferlini Incorporation of organics/inorganics nanocomposites in coatings of high environmental and 168 mechanical resistance J. Pimenta, C. Carneiro, A. Mendes, J. Moniz, F. Oliveira, F. Magalhães 152 Zenithal Illumination - from Alvar Aalto to Álvaro Siza's work Mafalda Rangel Campos

Contemporary condition: ‘new’ relationships between architecture and city

Mariana Alves1

1 Faculty of Architecture, University of Porto, Portugal.

The city is the real field of action of the architect. The architect of today doesn’t work on a tabula rasa, but on a complex structure of voids and fulls. The architecture of the city has always constituted a great challenge to the architects through the complexity that is directly associated with. It has a primordial role in the construction, transformation and identity of the cities. The thesis is structured in six chapters organized by case study, author and theme. In each chapter there is an approach of the architect projectual strategy, an analysis of the case study that includes an introduction, a characterization of the urban space, the environment context, the morphology and a critical synthesis of the author. In the introduction some remarkable examples of History, remind us the power of the architecture of the city in the development and construction of the urban space. The thesis focuses the impact of iconographic buildings in the contemporary city. Guggenheim Museum, in Bilbao, by Frank Gehry is an example of the catalyser effect of the architecture. The intervention has started an important revitalization in the city, spreading out a powerful energy through the titanic forms. Casa da Música, in Porto represents the objectual strategy. The formal contrast and the scale rupture that characterize the building make part of the conceptual approach of Rem Koolhaas – the architecture as a global product for a mass culture and for a metropolitan territory in a constant mutation. The Center Georges Pompidou, in Paris, is mainly an ideological building. Its complexity and contradiction come out through different directions as far as the relationship with the urban context and the architectural language concerns. On one hand, the approach to the city through the square, having as reference the dynamic italian square; on the other hand the technological language with references in Archigram’s projects. The impact of the tower typology in the urban space is developed with Burgo Building, in Porto and Seagram Building, in New York. The two towers mark the landscape through the abstract and synthetic value of the architectural system. With Burgo Building, Souto de Moura wants to build “a precise landscape”, a scenography and a specific abstract relationship with the place. The Galician Center for Contemporary Art and Serralves Museum are two buildings in which Siza reinforces the concept of architectural continuity. There are constant references to the importance of the context and the place identity. The shape of Siza’s buildings wants to participate in the shape of the city, wants to establish an order of continuity. The Tate Modern, in London, is a building where are present Rossi’s teachings about the permanence of the shape and the structuring value of the architecture of the city. Herzog & de Meuron operate surgically in the pre-existence reanimating the old industrial building as an important urban forum. All the study cases identify a place, transform the urban landscape and try to give answers to the complexity of the territory. They want to fix a time in the cities, to establish new polarities, to essay through the architecture prospective and erudite answers; add sensuality and intelligence to the urban space.

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Form and matter in the work of Rafael Moneo

Rui Gonçalves*

*Architect graduate from Faculty of Architecture, University of Porto, Portugal.

This investigation was undertaken in my final graduation thesis. It’s starting point is the attempt to define critique instruments, essential for the understanding of architectural practice as an articulate process of knowledge built on the relations between three crucial moments: design, construction and critique. From the observation of the problem of construction in architecture, the focus centers on the concept of Art-of-building (from the german baukunst) to define a group of principles for an interpretation of the work of Rafael Moneo (1937-). The main objective is to understand the relationship between the generative principles of the architect's work and the poetic speech that is inherent to the concept of tectonics, which refers to the artistic origin of architectural forms. Standing before Moneo's intellectual awareness (drawn and written) which defines itself by architectonic continuity, I think it is possible to establish a relevant articulation between his work and the concept of tectonics. Both trajectories lead to the essence of built forms and architecture. Moneo's understanding of construction as comprehension and, by consequence, as explanation of architecture is present since his first works, taking as an example the Gomez Acebo house (Madrid, 1966-68). Simultaneously, the expressly classical feeling of large masses that his works evoque is concretized in a narrative composed by elements that explore the ambiguous relation between construction and language, important for the discussion about the generative principles of architectural form. From his theoretical approach the text "Paradigms of XX century: the nineties, between fragmentation and compacity"1 (1999) stands out. It reclaims the compact in a continuity perspective and presents the sketch of a project instrument of crucial importance. The themes now identified find affinities with the ideas of Gottfried Semper (1803-1879), Kenneth Frampton (1930- ), Aldo Rossi (1931-1997) amongst others. Not worried about finding a direct connection between the architect and the various interpretations of the term in the course of history, tectonics in the work of Moneo should be understood as a critical matrix, as a different approach not only to the interpretation of his work and the support of his speech but also as a new way of looking at contemporary architecture. The reflexion about his speech and his built work tries to clarify the connection between architectonical forms and the concept of art-of-building, not in the sense of a direct identification but towards the characterization of the limit that separates the concepts of architecture and tectonics, trying to understand the interaction between spatial and formal thought and the notion of matter in the architectonic narrative that he establishes. ORAL References: [1] Moneo, R. (1999), "Paradigmas fin de siglo: los noventa, entre la fragmentación y la compacidad", Arquitectura Viva, 66, 17-24. [2] Gonçalves, Rui, Forma e matéria na obra de Rafael Moneo, Prova Final de Licenciatura em Arquitecura, Faculdade de Arquitectura da Universidade do Porto, 2007

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Architecture and climate. Site geography: Luanda and the work of Vasco Vieira da Costa

M. Quintã 1, J. Quintão 2

1 Faculty of Architecture, University of Porto, Portugal 2 Faculty of Architecture, University of Porto, Portugal

The present abstract refers to a Final Thesis for the Degree in Architecture of FAUP, which intended to perform a study of architect Vasco Vieira da Costa’s work. The journey to Luanda allowed to recognize the climate adaptation as a main principle in the work of this architect inducing, this way, a first investigation about the climate components. The first chapter intends, precisely, to prepare the investigation of the study case, specifying gradually the subject of the thesis, and that’s why it firstly broaches in a wide way the eternal confrontation between architecture and site, and then act particularly in the architectural intervention on tropical climates. After this first introduction, and also after the acknowledgment of the climate data of Luanda, the observation of the works focus particularly on their climate adaptation, using, above all, the information gathered during the journey. This way, a special attention is paid to the precisely orientation of the buildings in order to evaluate the importance of the shading and ventilation mechanisms used according to the sun and the main winds behaviour. It was intended to evaluate the configuration and dimensions of the architectural elements, inquiring their functional or poetic purpose. It was repeatedly observed that the use of correspondence between the climatic conditions of Luanda and the buildings of Vieira da Costa was constant along the architect’s work. To each project, Vieira da Costa used his principles; only facing a specific circumstance each solution was developed as a result of the program and the particular position in the territory. The unique person of Le Corbusier was decisive in the education period of Vieira da Costa, and his influence in the Angolan architect’s work will always be distinctive and permanent. And yet, the rational forms present in his works are not indifferent to the city where they are settled. Furthermore, relationships of continuity between the buildings of Vieira da Costa and the urban context are frequently established. Although the architect has defined the “solução de compromisso” [1] as the ideal setting - the half way between the ideal orientation to the sun and to the main winds - Vieira da Costa usually uses the city as a starting point.

[1] Vieira da Costa, V. (1969), Breves Considerações sobre Urbanismo Tropical em Zonas Rurais, in “Segundas Jornadas de Engenharia e Arquitectura do Ultramar”, Comunicações, vol.4, Luanda

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Crises e Reformas: o colonialismo ilustrado na Capitania de São Pauloe os esforços de modernização do Governo Morgado de Mateus (1765-1755)

Joana Salém Vasconcelos, Vera Lúcia Amaal Ferlini

Universidade de S. Paulo, Brasil

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Incorporation of organics/inorganics nanocomposites in coatings of high environmental and mechanical resistance

J. Pimenta1, C. Carneiro2, A. Mendes3, J. Moniz4, F. Oliveira2, F. Magalhães3

1 Rede de Competência em Polímeros, Porto, Portugal. 2 CIN – Corporação Industrial do Norte, S.A., Maia, Portugal 3 Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal. 4 Resiquímica – Resinas Químicas, S.A, Lisboa, Portugal

Inorganic fillers (metal oxides included) have been traditionally used in organic coatings, conferring improved mechanical and physicochemical properties. In recent years, the ability to produce inorganic fillers in nano-sized dispersible dimensions has attracted new attention over an otherwise well established field. Indeed, the performance of the inorganic-organic composites is optimized when the fillers are present in dimensions below 100 nm (nano-size). However, this is only effective if the materials are uniformly dispersed throughout the final coating film. When dealing with water based coatings, obtained from polymeric emulsions, it is important to ensure that the inorganic nanoparticles are well dispersed, either through encapsulation within the polymer particles or dispersion in the aqueous phase. The first situation may be the most desirable, since it should lead to a more uniform particle distribution in the coalesced film. One of the main motivations for this research is the development of enhanced intumescent paints (which expand when heated, creating a barrier to heat transmission and therefore protecting the substract from the high temperatures occurring during a fire [1]). Our work has been focused on incorporating nano-sized silica (SiO2) particles in vinylic emulsions. The initial approach consisted is pre-dispersing powdered nanosilica in the colloidal solution that is the starting point for the emulsion polymerization reaction. Two types of nanosilica were selected: Sil-1 (with surface treatment) and Sil-2 (without surface treatment). For Sil-1, some methods of dispersion have been studied in order to avoid suprananometric agglomeration in the aqueous phase: Cowles agitator, ultrasound and bead milling. The later gave the best results. After polymerization of the vinylic monomers, TEM microscopy indicated that there was silica present in the surrounding aqueous phase, probably stabilized by the colloid, and encapsulation by the polymer could not be confirmed due to the opacity of the polymeric particles. Concerning Sil-2, TEM analysis indicated that these nano particles tend to attach to the surface of the polymeric particles and encapsulation is improbable in this case (Fig. 1). Current work is dealing with promoting the encapsulation of Sil-1 in the polymer, by developing a colloid-free formulation, and the encapsulation of Sil-2, by using a comonomer with affinity to its surface.

Figure 1 – polymer emulsion particle surrounded by Sil-2 References: [1] – Nogueira J. L. (2006), Protecção passiva contra o fogo.

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Zenithal Illumination – from Alvar Aalto to Álvaro Siza’s work

Faculty of Architecture, University of Porto, Portugal, Mafalda Rangel Campos

This investigation tries to make an approach to the theme of Light Presence in Aalto’s and Siza’s architecture. It faces daylight as one of the major aspects, which influences their project’s process, in which light is a geographic variant. Which architectonic forms they have chosen to interpose between Man and Sun, so that a certain space becomes a library or a museum? A previous analysis of their most important creation makes it possible to understand how often the use of zenithal illumination is associated to public buildings. As a consequence, this study explores its presence in libraries, museums and exhibition spots. The structure of this investigation reflects its methodological process. It starts from the conceptual analysis of light in architecture as a way to clarify fundamental aspects of the theme, progressing towards the practical analysis of several public buildings. Chapters I and II explain the conceptual and historical study of the zenithal illumination, tracing a constructive evolution. Chapter III makes an approach to the trajectory of both architects and the cultures they are bound to, also including evolution of their work and individual exploitation of permanent concepts in their legacy. Chapter IV explores an analysis of selected works, establishing direct and indirect relations between them, in order to systemize space characters created by the zenithal illumination. Chapter IV shows that both architects have invented, re-created and re-used different daylight control systems, mostly using indirect and diffused lighting for libraries and museums. It was possible to systematize and group buildings according to the use of the same shape and therefore understand the chronological process the architects have gone through. It possible to conclude that the shapes chosen by Aalto and Siza to interpose between Man and Sun are not only directly connected to the existing daylight conditions. In their libraries and museums it is visible, that the process of shaping a roof is always a compromise between the light conditions of the place and the individual formal research that each architect has been developed.

Fig. 1 Viipuri Library- Reading room - Alvar Aalto Fig. 2 IT Faculty Library- Main reading Room-Santiago de Compostela - Álvaro Siza

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PARALLEL ORAL SESSIONS III – POS.III.3 THURSDAY, FEB. 21

II.3 - ARCHITECTURAL PERSPECTIVES Georgina Correia da Silva & Filipe Abrantes 195 Architectural Languages for the 21st Century N. Vieira da Costa, J. Quintão Exhibiting Architecture in Portuguese. Curatorial activity in Portugal pre- Trienal de Arquitectura de 189 Lisboa Mariana Pestana 213 The "Eclipse" of architecture D. Neves dos Santos, Prof. Manuel Mendes 124 Promenade to Corbusier L. Ribeiro da Silva, M. Mendes 206 Architecture: Studies and possibilities of transversalities. Space of Encounter – Herzog & de Meuron, Diller + Scofidio, Dan Graham, Lacaton & Vassal,

Sancho y Madridejos, Vito Acconci [1] Ana Pedrosa 219 Contact Points_Between Swiss and Portuguese Architecture Bruno M.V. Cardoso

Architectural Languages for the 21st Century

N. Vieira da Costa1 and J. Quintão2

1 Faculty of Architecture, University of Porto, Portugal. 2 Faculty of Architecture, University of Porto, Portugal.

This study pretends to be a comprehension of the present situation in architecture. For that, we proceeded to a distribution of the methods and languages present in contemporary architecture, by groups and in a hierarchic disposition, based on their formal, technical and conceptual similitude. This systematised classification corresponds to the taxonomic scheme presented in the study and constitutes its essential synthesis. The architectonic approaches explored in the study have been identified from the observation of works conceived or built between 2001 and 2005 published in the magazines L’ Architecture d’Aujourd’hui, The Architectural Review, Arq/a, Casabella and Pasajes. It is important to refer that it is not our intention to classify the architects, only the architectonic works in question. The study persists on the idea of architectural language by identifying witch formal, technical and conceptual rules are in the base of the diverse architectonic approaches. We also try to clarify the genesis for each architectural posture; in the sense of knowing what prevails, if the desire for tradition or that of reinvention. This method allowed us to determine the origins behind concepts and theories and to establish differences and similarities towards the dominant architectural concepts. Subsequently, the architectural concepts proceeding, synthesizing or contesting the prevailing traditions were grouped in several postures that maintain an internal coherence. Along the study, theories coming from sciences and arts’ fields are also taken into consideration. Consequently, here are described both the “meta-languages” (theories, concepts, utopias, etc.) and the methods and languages applied to the architecture in concrete. Therefore, this study doesn’t consist on a description of architectonic works; it establishes a dialectical relation between the theoretical concepts and the concrete languages. Key words: methods and languages; contemporary architecture; systematised classification; genesis; sciences; arts.

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Exhibiting Architecture in Portuguese. Curatorial activity in Portugal pre- Trienal de Arquitectura de Lisboa

Mariana Pestana1

1 Faculty of Architecture, University of Porto, Portugal.

Despite the increase in architecture exhibitions over the last years, and the granted power of architecture exhibitions as definers of concepts and movements, little research has been done on this theme in Portugal. It seemed to me then relevant to study architecture exhibiting in Portugal. Its contemporaneousness, allied to the fact that it represents a “compilation” of architecture exhibitions in Portuguese, made the first Trienal de Arquitectura de Lisboa my chosen case study. Defining the role of the curator, the contribution of exhibitions to Architecture and the communication for different audiences were the main questions raised. I interviewed people related to this activity in Portugal, researched in archives and libraries, and worked as a volunteer at the Trienal. With the gathered information I assembled an analysis board where the Portuguese curatorial activity until Trienal was arranged. The act of displaying is present in occidental society, associated with processes of identity construction, historical narratives and collective memory. Within these exercises, the curator’s importance stands out. And what about the architecture display in particular? The curator presents a partial version of reality, situated in a specific time and space. Architecture’s immobile nature means that its representation in exhibition is achieved with its content absent. The curator’s performance, as mediator between public and content plays, then, there, an especially determining role. Exhibition, as part of Architecture, has had a fundamental role in its fashions’ and tendencies’ conceptual clarification. From laboratory to critical activity stage, the role of the exhibition may be experimentation, social intervention or education, among others. Architecture is represented through a codified language. Public accessibility to its contents becomes an inevitable question that raises questions about the pertinence of displaying conventional representative elements. The approximation to reality is not necessarily an objective. Architecture’s multiple dimensions - beyond mere built object, as a social interface element, for example, as well as its everyday presence as component of life, do give it countless directions in which to approach an exhibition. Trienal de Arquitectura de Lisboa opened a regular space for architecture presentation through different forms, among them exhibition. The 2007 event was organised starting with a team of curators, who were given the responsibility of “putting Portugal on the architectural map”. The curatorship of architecture’s does exist in Portugal. Therefore I believe it must be debated and discussed. It is important to understand what its objectives are, its specificities, and, of course, it must be practised - for and as Portuguese architecture. To discuss exhibitions’ contribution to architecture and to analyse communication with different audiences are questions that I will approach.

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The “Eclipse” of architecture

D. Neves dos Santos1, Prof. Manuel Mendes (tutor)2.

1 Faculty of Architecture, University of Porto, Portugal. 2 Faculty of Architecture, University of Porto, Portugal.

The concept of eclipse applied to architecture emerged in the course of a study of an influent house –C. S. H. #8 (1945-1949), the only house designed by (and for) Charles and Ray Eames, promoted by Arts&Architecture magazine within the Case Study House Program. Marked by a doubt between two different solutions for the same site, were left contradictory documents about the Eames idea of architecture. If on the one hand it says that architecture could be a virtuous composition of forms legitimated by a style directly imported from the authors of the modern movement; on the other hand, caused by contingents (or taking the most out of them), one can see that Eames took a different position in view of architecture because, on the built house, the most sublime moments of architecture were recorded as a form’s eclipses and never as a form’s exhibition. Motivated by this hesitation, we found space to question and propose a new statement in view of most elementary matters underlying the architecture exercise: which is the subject/purpose of our discipline? What kind of knowledge could inform the conception according to those purposes? How can architecture, under that purpose, be represented? How can architecture be materialized? Nevertheless, the analysis’s field to which theses question are linked is not limited by dates or geographic places where some historical incident has happened. The criteria that bound the fields of our action is a criteria made by the “time” of daily life architecture. No one more than Charles and Ray Eames left such a large influence by an unique collection made by a million of fragmentary and causal records exposing the ordinary moments of their architecture mixed with their everyday life. We are interested in facts that are out of the famous moment of inauguration or when Julius Shulman was at Eames House to make the famous photo that everybody recognizes as CSH #8. Looking at that house through images of a time that is a time of living, we realise that architecture as an object was softly disappearing from photos, drawings or other records of that house, as we can confirm on House-after five Years of Living (1954) - the Eames`s most relevant document about the house ). With the passing of time, the object of a house looses the leading role representing architecture and it is replaced by “the things” that this object made possible, because for Eames, architecture is something beyond it’s projects, it’s records and it’s objects. Architecture is a “private landscape”. Through an Eames eye, we describe the possibility: architecture could be an immaterial product, more influenced by actions than proportion. Consequently, we found an apparent contradiction on architectural representation: drawings, models, renders infer a constructive description of landscape that is always on movement, evanescent…and made of spatial story. So, the representation betrays the reality in the same way that representation betrays architecture. This special attention given to architecture, beyond the inauguration or “photo days”, has an apparent price: became invisible. Eclipsed. This invisibility is not humble because the ambition is simultaneously unmeasured: to find happiness through experiencing the space that we project or through observing water draining into the school’s courtyard. Isn’t it Mr. and Mrs. Eames?

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Promenade to Corbusier

L. Ribeiro da Silva 1 and M. Mendes2

1 Faculty of Architecture, University of Porto, Portugal. 2 Faculty of Architecture, University of Porto, Portugal.

“Promenade to Corbusier” proposes to analyze the “quality of space” in the work of Le Corbusier, quality only recognizable through the direct “experience of architecture”. In an early stage of the investigation, it was believed that the “quality of space” would be entailed to the concept of “Promenade Architecturale”, enunciated by Le Corbusier himself. However, the further development of the work has resulted in the understanding of the “Promenade Architecturale” as a fragment of a broader domain: the “Promenade to Corbusier”. “Promenade to Corbusier” is structured upon the recognition, in corbuserian architecture, of a “conception field” – “traversing” [1] – and a “subject” that performs this action – the “traverser” [2]. “Traversing” is acknowledged as an event of universal inevitability: physic inevitability, biological inevitability, religious inevitability. “Traversing” is acknowledged as existential synthesis. Four propositions of “traversing” are recognized, according to the condition of the “matter” and the modes of its manipulation. To each proposition of “traversing” corresponds a state of “traverser”. The first proposition of “traversing”, “Body’s Conduction”, concerns a field of architecture where the coordination of movement is directly related to the “quality of space”. This is the field of architecture concerned by the concept of “Promenade Architecturale”, starting point of the investigation. The “traverser” moving through the “inert matter”. The second proposition of “traversing”, “Sight’s Conduction”, identifies the “traverser” in the process of slowing down and seeks for the recognition of the decreasing motion’s space. “Sight’s Conduction” concerns the possibilities of the “promenade” by framing the visual field of the “traverser”, in a fixed spatial position, through the “inert matter”. The third proposition of “traversing”, “Retina’s Stimulation, the Natural Matter”, identifies the “traverser’s” spirituality and seeks the recognition of the abstraction’s space. “Retina’s Stimulation, the Natural Matter” concerns the possibilities of the “promenade” by managing the motion of light, “natural reactive matter”. The forth proposition of “traversing”, “Retina’s Stimulation, the Artificial Matter”, concerns the recognition of the increasing possibilities arisen by civilizational development, in the origin of the synthesis of “artificial reactive matter” – from the “electric” to the “electronic” – with the potential to operate a revolution of both “quality of space” and state of “traverser”. «The verb is to be» [3].

References: [1] “percorrência”. [2] “percorrente”. [3] Costa, M. E. (1997), Lucio Costa – Registro de uma Vivência, Empresa das Artes, São Paulo, pp. 4.

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ARCHITECTURE: STUDIES AND POSSIBILITIES OF TRANSVERSALITIES. Space of Encounter – Herzog & de Meuron, Diller + Scofidio, Dan Graham, Lacaton & Vassal, Sancho y Madridejos, Vito Acconci [1]

Ana Pedrosa

Faculty of Architecture, University of Porto, Portugal.

This study seeks a personal idea of a contemporary architectural practice, considering above all its artistic potential of generating meaningful events in space, rather than the imperative to construct. The study object is the production processes of architectural poetry, having as key strategy the transversality between distinct disciplinary practices and between discourse and practice. This interdisciplinary is not constraint to other artistic activities. Nevertheless, the work addresses particularly the practices of installation and performance, for its manifestations by actions towards space or body and therefore focusing more on the effects prompted and less on objectual character of the work of art. To establish coincidences or interferences between enclosed disciplines is not considered as a value for itself. What is at stake is the reinvention of an architectural practice through the redefinition of its limits as a discipline, in its objects, in its tools, in the author’s role, in the inhabitant’s role, in its relation with place. The idea of architecture, outlined along the essay, as an artistic medium of intervention, results of the conjuncture of the following: an idea of the qualities of its meanings – seen as unpredictable, wandering, subjective – through the notion of indetermination; an idea of the inhabitant’s active and creative role in the construction of the experience of architecture, through the notion of event; an idea of art as a potency of the real , through the notion of virtual; an idea about the identity limits of the subject, as well as of the architectonic object, through the notion of body. [1] Graduation Essay – 2007. Supervisor Teacher: Manuel Mendes, Arguer Teacher: Manuel Botelho, Jury: Rui Braz. The early study issues were induced by the internship at the Atelier Sancho y Madridejos – Madrid, 2003-04 – who’s practice is inspired in the work strategies of artists as E. Chillida and J. Oteíza; and the participation in the Workshop Espaço do Encontro (Space of Encounter), under the orientation of Jean Philippe Vassal – Porto, 2005, and consequent public space intervention at the FIMP 2006 – Porto International Puppets Festival.

Main theoretical bibliographic references (summarily identified in order to acomplish page layout): Deleuze, Gilles. Conversações 1972-1990,(1990); Foucault, Michel. “Des Espaces Autres” in Dits et Écrits IV 1954- 1988,(1957); Benjamim, Walter. “La obra de Arte en la Época de su Reproductibilidad Técnica,”(1936); Cruz, Teresa. “Posfácio” in O Pintor da Vida Moderna,(1993); Moure, Gloria (editor). Behind the facts – Interfunktionen 1968-1975,(2004); Ortega y Gasset, José. “La Deshumanización del Arte,” in La Deshumanización del Arte y Otros Ensayos de Estética,(1925); Conde, Yago. Arquitectura de la Indeterminación,(2000); Guilheux, Alain. Architecture Action – une Architecture Post-Théorique,(2002); Leatherbarrow, David. Uncommon Ground – Architecture, Technology, and Topography,(2000); Solà- Morales, Ignasi. Territorios,(2002); Solà-Morales, Ignasi. Diferencias – Topografia de la Arquitectura Contemporánea,(2003) ; Teyssot, Georges. “The Mutant Body of Architecture” in Flesh – Architectural Probes, (1994).

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CONTACT POINTS_BETWEEN SWISS AND PORTUGUESE ARCHITECTURE

Bruno Miguel Varejão Cardoso

Faculty of Architecture, University of Porto, Portugal.

The current society is in constant and fast transformation. With the easy and fast displacement of people and information a new confluence becomes possible and thus confrontation between different cultures. The borders of the knowledge extend to a global level, subverting the concepts of time and space, creating bonds of stronger union and proximity between the different world-wide cultures, in a process of constant definition of the European identity, based in a strong interaction between the globalization and the various local trends. This reaction marks deeply the practical of the new generations of young Portuguese architects. They developed their formation with values based on a culture of openness to Europe, in the appearance of a new society of consumption and information, in a search for new concepts that reflects a disciplinary osmosis of architecture with other artistic movements and trends, in conciliation with the important legacy of the 50’s generation, opening space for reflection and debate on the past and present of the Portuguese architecture. This is reflected in a quality production with new vectors of action and growth, with a new dynamism and concept, and it marks a time of change in the architectural scene. The Swiss architecture reveals conceptual and minimal issues, with a constructive rigour and a formal simplicity, and modern concepts re-interpreted. It became a great reference for young Portuguese architects, for revealing a reconciliatory attitude between traditional concepts and broad cultural transformations of contemporary society, maintaining a simple language and an honest construction, readily identifiable with the Portuguese tradition. While trying to flee from a predominantly complacent and uncritical culture, the young architects who were deploying in the Portuguese architectural scene, kept the search for an adequate response to a local context stopped in time, faced with new demands of a confluence for a new global and more dynamic context. These new generations demonstrate perseverance, willingness and dedication, with a strong ability to explore the risk, using the marginal contexts to avoid stagnation and conformism, looking to assert themselves with an attitude between resistance and commitment. This new awareness includes the recognition of our identity, traces in evolution, a change in the traditional approaches, influences, and references, that will transform the context of the Portuguese architecture and allow a conscious and positive metamorphosis of our cultural specificity in loss, and a slow transformation of the discourse in the search for our identity as Portuguese, as Europeans.

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PARALLEL ORAL SESSIONS IV – POS.IV.1 THURSDAY, FEB. 21

IV.1 - GREEN CHEMISTRY & ENVIRONMENT Elisa Keating & Jorge Teixeira 58 Levels of Butyltins in Sediment and Suitability of Salt March Plants for Rhizoremediation P. N. Carvalho, P. N. R. Rodrigues, M.C.P. Basto, M.T.S.D. Vasconcelos Implementation of automatic methodologies for quality control of pharmaceutical formulations 48 containing allopurinol based on biocatalysts C. I. C. Silvestre, M. A. Segundo, M. L. M. F. S. Saraiva, J. L. F. C. Lima Azo-dye Orange II degradation by Fenton-like reaction using as catalysts transition metals in carbon 63 supports Filipa Duarte, F.J. Maldonado-Hódar, Luis M. Madeira 86 Monitoring of Biological Remediation of a Soil Polluted with Petroleum Hydrocarbons E. Monteiro, M. N. Couto, M. T. S.D.Vasconcelos Isolation and characterisation of a novel Planctomycete bacterium related to Isosphaera group from 108 a freshwater aquarium Joana Bondoso and Olga Maria Lage 227 Intracellular Gene Regulation via Engineered Nanolipoblockers for Inhibition of Cholesterol Uptake Ekta Patel, Prabhas Moghe

Levels of Butyltins in Sediment and Suitability of Salt March Plants for Rhizoremediation

P. N. Carvalho1,2,*, P. N. R. Rodrigues1,2, M.C.P. Basto1,2 and M.T.S.D. Vasconcelos1,2

1 Department of Chemistry, Faculty of Sciences, University of Porto, Portugal. 2 CIMAR/CIIMAR – Centre of Marine and Environmental Research, University of Porto, Portugal. * [email protected]

In recent decades organotin compounds have been extensively used in agriculture and industry, as biocides and polymer stabilizers. Since the 1980s contamination of waters and sediments with tributyltin (TBT), as a results of its application in paint for boats as an antifouling, has been associated to a variety of unexpected malformations and other effects in non-target marine organisms. Therefore, it is necessary to control the application of the legal impositions, ban of TBT from the global fleet, as well as the release of TBT and its degradation products from contaminated areas, as well as study and develop remediation techniques. For the accomplishment of these purposes, accurate and very sensitive analytical methods are required for quantifying trace levels of butyltins (BTs) in various sample matrices. Very recently, a method was developed and validated using headspace solid-phase microextraction as an automated pre-concentration/extraction process followed by gas chromatography separation coupled to tandem mass spectrometry detection. The method was efficiently applied to the determination of TBT and its degradation products dibutyltin and monobutyltin in sediments from Sado River estuary [1]. Concentration values of TBT over the range established by the OSPAR [2] as provisional ecotoxicological assessment criteria that should be used to identify areas of concern were found in the Lisnave area. To investigate the capability of the salt marsh plant Halimione portulacoides to promote BTs degradation, plants were transplanted ex-situ into several pots with sediment from Lisnave area. Identical pots without plants were used as controls. Monitoring of BTs in those sediments [1] was carried out. It was observed that, in the absence of the plant, natural degradation of TBT along eight months was negligible. In contrast, a significant decrease of contamination occurred in the presence of plants, denoting rhizoremediation, despite the low belowground biomass of H. Portulacoides used in the tests. Further studies are in progress, using higher amounts of biomass per experiment, with the purpose to investigate whether the plant participates directly in the BTs degradation, namely incorporating TBT or its metabolites in plant tissues, or it only promotes degradation by sediment microorganisms from the rhizosphere (environment around roots).

References: [1] Carvalho, P.N., Pinto, L.F., Basto, M.C.P. and Vasconcelos, M.T.S.D. (2007), Headspace solid-phase micro-extraction and gas chromatography-ion trap tandem mass spectrometry method for butyltin analysis in sediments: Optimization and validation, Microchem. J., 87, 147. [2] Joint Meeting of the Oslo and Paris Commissions (1997), Oslo and Paris convention for the prevention of marine pollution, Annex 6, Joint Meeting of the Oslo and Paris Commissions, Brussels. Acknowledges: Project POCTI/CTA/48386/2002 financed by FCT, Portugal

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Implementation of automatic methodologies for quality control of pharmaceutical formulations containing allopurinol based on biocatalysts

C. I. C. Silvestre, M. A. Segundo, M. L. M. F. S. Saraiva and J. L. F. C. Lima

REQUIMTE, Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal Email: [email protected]

Allopurinol is widely used in the management of hyperuricaemia, due to its capacity to inhibit xanthine oxidase [1]. Xanthine oxidase, a metal flavoprotein, is the last enzyme in the pathway for the degradation of purine derivatives and therefore the rate limiting step in purine metabolism. Its main role is to oxidise hypoxanthine and xanthine to uric acid. In the present work, two automatic sequential injection analysis methodologies were developed for the enzymatic determination of allopurinol. Both methodologies were based on the inhibition by allopurinol of the reaction catalysed by xanthine oxidase in vivo, assessed by spectrophotometric detection of the reaction product (uric acid) at 295 nm. Furthermore, one methodology was based on a single readout at a fixed-time after reaction while the other was based on the decrease of the rate of formation of uric acid, evaluated by monitoring the absorbance change during a pre-set time interval. Both methodologies allowed fast quantification of allopurinol in pharmaceutical samples (n = 8), providing results that were not significantly different from those obtained by the British Pharmacopoeia procedure. The determination frequency was 15 and 34 determinations per hour for the fixed-time and kinetic method, respectively. The developed methodologies allowed the automation of the determination of allopurinol with the purpose of its quality control, implementing Green Chemistry concepts [2] in the Analytical Chemistry area. In this particular application, low quantities of reagents (µL range) were used and a low volume of effluent was produced (< 5 mL) per assay. Furthermore, the utilization of a biocatalyst for this determination, instead of rather toxic reagents, also makes it more environmentally friendly.

References: [1] Borges, F., Fernandes, E., Roleira, F. (2002), Progress towards the discovery of xanthine oxidase inhibitors, Current Medicinal Chemistry, 9 (2), 195-217. [2] Prado, A. G. S. (2003), Química Verde, os desafios da química do novo milénio, Química Nova, 26 (5), 738-744.

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Azo-dye Orange II degradation by Fenton-like reaction using as catalysts transition metals in carbon supports

Filipa Duarte a, F.J. Maldonado-Hódar b, Luis M. Madeira a

a LEPAE, Departamento de Engenharia Química, Faculdade de Engenharia da Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal b Departamento de Química Inorgánica, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain

Pollution of water caused by industries is a serious problem in the developed countries and its elimination is a great challenge. For instance, to destroy synthetic dyes is very hard because they are usually stable compounds and often non-biodegradable, formed by long organic molecules. Advanced chemical oxidation methods can be effective for their degradation as they can result in almost complete mineralization in a cheap and easy way. One of the most well-known methods is the oxidation with Fenton’s reagent, where hydrogen peroxide reacts with iron (or with another transition metal catalyst) in acidic medium, leading to the production of powerful radicals (HO•), which are able to attack the organic compounds. This reaction can be carried on in a homogeneous or heterogeneous system. The first process is not adequate because it leads to a very high metal concentration in solution, ca. 50-80 ppm, while European directives don’t allow more than 2 ppm of Fe, for instance, in discharged water [1]. Thus, heterogeneous reaction has been carried out using transition metals (Fe, Co, Ni, Cu, Mn, W) as active phase deposited on carbon aerogels. Two approaches were used to prepared the catalysts: in the first case, the metal precursors were introduced in the starting solution of the polymer precursors, resulting in metal-doped carbon aerogels; in the second case, metals were added by impregnation on two carbon aerogels previously prepared, one mesoporous (support M) and another microporous (support AS-3). The porosity and dispersion of the catalysts was evaluated by different techniques: N2 adsorption, mercury porosimetry, TG, XRD, HRTEM and SEM. The dye degradation experiments were performed in a slurry batch reactor, using a 0.1 mM orange II (model compound) solution, as well as 6 mM of hydrogen peroxide, at 30 ºC and pH = 3. The absorbance was continuously read by a spectrophotometer and the metal leaching quantified by atomic absorption. The best performance was obtained with the Fe, Co and Ni catalysts, although the latter was not so active and the second provided high leaching values. It is noteworthy that the Fe-doped catalyst showed the lowest leaching. When employing impregnated catalysts, the use of mesoporous supports leads to more active catalysts. The AS-3 support has a very high adsorption capacity, not being relevant for catalytic purposes. Comparing doping and impregnation techniques, it is observed that impregnation is more efficient probably because metals (Fe or Co) are more accessible to the dye molecules, in spite the high porosity of metal-doped carbon aerogels. The combination of the mesoporous support M with a highly active metal as Fe provided an efficient and promising catalyst for practical applications, with complete decolourisation of the solution in 2-3h employing soft oxidation conditions, with leaching values below 2.5%. References: [1] Sabhi, S. and Kiwi, J. (2001), Degradation of 2,4-dichlorophenol by immobilized iron catalysts, Water Research, 35 (8), 1994-2002.

86

Monitoring of Biological Remediation of a Soil Polluted with Petroleum Hydrocarbons

E. Monteiro1*, M. N. Couto1,2, and M. T. S.D.Vasconcelos1,2

1 Faculty of Sciences, University of Porto, Portugal. 2 CIMAR/CIIMAR – Centre of Marine and Environmental Research, University of Porto, Portugal *[email protected]

Due to several human activities many petroleum hydrocarbons (PHC) have been introduced in soil and sediments. Much PHC, particularly the less volatile ones (LVPHC), are persistent and can be tightly sorbed into soil particles, decreasing PHC bioavailability.

Biological remediation is a low cost way for recovering polluted soil, which rehabilitates soil structure. A project is in process to survey, as exhaustively as possible, the main factors that influence the efficiency of biological degradation of LVPHCs assisted by vascular plants (bioaugmentation and rhizoremediation). Plants well adapted to coastal areas and capable of living in soils with relatively high salinity were chosen. This pilot study is carrying out ex-situ, in polluted soil from a petrol refinery. At the beginning and during the study, the LVPHCs contents are determining both in soil and lixiviate, by using spectrophotometry [1], after suitable PHC extraction from the samples. The role of the under-graduated student in the project is the optimization of analytical methods for an expeditious monitoring of the efficiency of PHC degradation.

This project provides a real mesocosmos study, in a field with specific characteristics, and it is expected that in a middling/long time it will give clear conclusions about suitable conditions for decontamination of either the global area of the refinery or other sites with similar contamination.

In this communication, the first results of the project will be presented and discussed.

[1] Liste, H-H and Felgentreu, D. (2006), Crop growth, culturable bacteria, and degradation of petrol hydrocarbons (PHCs) in a long-term contaminated field soil, Applied soil Ecology 31, 43- 52. Acknowledgements: To FC&T for the Ph.D. scholarship of M. N. Couto (SFRH/31816/2006). To Refinery of Oporto (GALP Energy) for financial support. Logistical support by J. Amorim, from Refinery of Oporto and technological advices and logistical support by I. Teixeira, R. Salé and E. Bernardes from Berka – Engº E. Bernardes, Lda and M. I. Caçador from Oceanographic Inst. of Sci. Fac, L.U.

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Isolation and characterisation of a novel Planctomycete bacterium related to Isosphaera group from a freshwater aquarium

Joana Bondoso1,2 and Olga Maria Lage1, 2

1 Departamento de Botânica, Faculdade de Ciências, Universidade do Porto 2 CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental

Planctomycetes are an intriguing and fascinating monophyletic phylum of the domain Bacteria (3). Members of this group are characterised by budding, absence of peptidoglycan in the cell wall and cellular compartmentalisation. This contradicts the traditional dichotomous organization of cellular life in prokaryotes and eukaryotes and places them at the centre of the evolutionary microbiology for the search of the last common ancestor of the domains of life (1, 2). Their ubiquity and relative abundance has been revealed by recent molecular microbial ecology techniques. Knowledge of this group is very limited due to the relatively few species grown in pure culture, thus the great importance of isolation and cultivation studies for a better understanding of its ecological role. Here we report the isolation in pure culture and the characterization of an Isosphaera- like bacterium from the sediment of a freshwater aquarium. After about 3 months incubation of the sediments in modified selective (200μg.mL-1 ampycillin, 500μg.mL-1 streptomycin and 20μg.mL-1 cycloheximide) IMC and PEO3 media at 20ºC in the dark, light pink colonies appeared. CARD-FISH analysis of the sediments revealed the presence of round-shaped PLA46 (Planctomycetes 16S rRNA oligonucleotide probe) labeled cells. Cells are about 2 µm in diameter, normally appearing as individuals, but having the possibility to form small chains of 2-5 cells. Division occurs by budding. Scanning electron microscopic analysis revealed the presence of the characteristic crateriform structures and a compact matrix linking the cells, and ultrathin sections (Transmission Electron Microscopy) of the cells showed the DNA surrounded by an electrondense area. Nutritional requirement studies showed that the characteristics of the colonies, namely consistency and color, are media dependent. Isosphaera-like OJF2 can utilize glucose, rhamnose and melibiose as the sole carbon source and liquefies gelatin. Optimal growth occurs at 35ºC and 0.35% NaCl. The almost complete (1480bp) 16S rDNA sequence showed its affiliation to the Isosphaera group (Isosphaera-like strain OJF2, GenBank accession number DQ986200), with the highest similarity to uncultured bacteria from environmental samples. The closest cultured relative (96%) is Isosphaera sp. Schlesner 666 and it shows only 94% similarity to I. pallida, the type species. This fact associated with a diverse morphology, indicates that this strain could potentially be a new genus of the order Planctomycetales. References: 1. Fuerst JA. 1995. The planctomycetes: Emerging models for microbial ecology, evolution and cell biology. Microbiology 141: 1493-506 2. Fuerst JA. 2005. Intracellular compartmentation in planctomycetes. Annual Review of Microbiology 59: 299-328 3. Garrity GM, Holt JG. 2001. The roadmap to the manual. In Bergey’s manual of systematic bacteriology, ed. DR Boone, RW Castenholz, pp. 119-66. New York, N.Y.: Springer

88

Intracellular Gene Regulation via Engineered Nanolipoblockers for Inhibition of Cholesterol Uptake

Ekta Patel, Prabhas Moghe

Chemical and Biochemical Engineering Department, Rutgers The State University of New Jersey

Atherosclerosis, the pathogenic accumulation of low-density lipoprotein (LDL) within the blood vessel walls, is a leading cause of death worldwide. While systemic therapies inhibit LDL synthesis, our approach involves the localized management of oxidized LDL within the vascular wall through the use of anionic nanolipoblockers (NLBs) and macrophage-targeted drugs. We have shown that NLBs reduce foam cell formation by receptor blocking, a hallmark of atherogenesis; however, their effect on cellular gene expression is unclear. We investigated the effects of NLBs on genes regulating atherogenesis and inflammation, with the goal of identifying the most antiatherogenic NLB configurations.

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PARALLEL ORAL SESSIONS IV – POS.IV.2 THURSDAY, FEB. 21

IV.2 - LIFE & WORK & SOCIETY Manuela Aguiar & Albino Lima 52 The relativity of happiness: Portugal and the EU J.P. Fique, D.A. Sá 117 Natural resources course B. Magalhães, C. Mota, F. Martins, J. Silva, L. Fernandes, N. Teixeira, P. Monteiro, P. Santos, R.

Oliveira, T. Teixeira, V. Oliveira 148 Processes of corruption in Portugal: results of an exploratory research R. Faria, C. da Agra 223 Desistência do tratamento de enurese: avaliação dos motives alegados e de variáveis associadas Carolina Ribeiro Bezerra de Sousa, Edwiges Ferreira de Mattos Silvares 30 Popular culture and the formation of the working class J. Aguiar A troubled legacy. Changing strategies of social transmission and patterns of identity production 33 among industrial workers. Bruno Monteiro

The relativity of happiness: Portugal and the EU

J.P. Fique1, D.A. Sá2

1Undergraduate student, Faculty of Economics, University of Porto, Portugal. 2Undergraduate student, Faculty of Economics, University of Porto, Portugal.

“Life is a progress from want to want, not from enjoyment to enjoyment” Samuel Johnson, 1776

What makes us happy? This everlasting question gained recently a good amount of attention from Economics. Until this unexpected focus from Economics, psychologists have been studying this matter, which became a valuable support for economic analysis of happiness. In the present analysis it will be considered the relation between happiness and income. The reference in this relation is the work developed by Richard A. Easterlin (1974, updated in 1995), the so called Easterlin Paradox consists in the “paradox of substantial real income growth in Western countries over the last fifty years, but without any corresponding rise in reported happiness levels” (Clark, A.E., Frijters, P., Shields, M.A, 2006)[1]. But, a second literature has emerged pointing that money does matter. The result of the reconcilement of Easterlin Paradox and this second literature is that happiness is relative, it depends on both external and internal reference points. Extending this theoretical result, we can consider as Kapteyn et al (1976) as previously argued, that reference groups include whole countries. We propose to question if the low rank of Portugal in terms of Life Satisfaction is a result of considering the European Union as a reference group. Hence, the low Life Satisfaction results would be a consequence of the difference between Portugal’s income and the supposed reference group. To do so, we will use data available from Eurobarometer. In terms of theoretical orientation, it will follow the perspective of “Clark, A.E., Frijters, P., Shields, M.A, 2006”[1] in close propinquity. This analysis the proposition of the application of a theoretical perspective to the understanding of happiness, which will be used in our ongoing research project “Happiness and the Human Development Index: The case of Lisbon and Porto”. Therefore, it will aim to favor a discussion about this topic and not present actual results. What we propose to do is identify in the Eurobarometer data results predicted by this line of thought.

References: [1] Clark, A.E., Frijters, P., Shields, M.A. (2006), Income and happiness: Evidence, explanations and economic implications, Paris-Jourdan Sciences Economiques, WP nº 2006-24. [2] Easterlin, R.A. (2001), Income and happiness: towards a unified theory, in The Economic Journal, 111 (July), 465-484. © Royal Economic Society 2001. Published by Blackwell Publishers, 108 Cowley Road, Oxford OX4 IJF, UK and 350 Main Street, Malden, MA 02148, USA

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Natural resources course

B. Magalhães, C. Mota, F. Martins, J. Silva, L. Fernandes, N. Teixeira, P. Monteiro, P. Santos, R. Oliveira, T. Teixeira and V. Oliveira

Department of Geography, Faculty of Arts, University of Porto, Portugal

A clear example of a rich country in natural resources is Angola, especially oil, but one of the problems these types of countries have is a large tendency for bad government, armed conflicts and weak action in terms of economic and social development: we could say that countries with fragile institutions have also little capacity to tolerate the destructive forces of corruption, nevertheless if these resources were properly used, Angola would be one of the richest countries of Africa. Our purpose is to prove that there is a correlation between population, political movements and natural resources. The used method is based on OCDE/AIE Report 2006, which analyses statistical data to elaborate a thematic cartography using the ArcGis 9.2 program, seeking to conclude that there is an historical relation between the three vector areas. With a relatively small population, only 14,5 million inhabitants, Angola should be capable to present indicators of well-being among the most developed countries in Africa, instead of that, war and bad government increase the oil waste. This way it becomes very interesting to analyze natural resources, demographic and political movements in Angola, including Cabinda´s area, once they origin many social and economic deficiencies. It becomes clear that the different political movements had as main purpose to control natural resources. Quality in administrating resources will probably be the most important factor to determine if Angola achieves a maintainable peace or if, it falls once again in conflict. There are two essential questions: - to know which are the government priorities concerning expenses; - the transparency in the administration of public finances. For several years, and for armed conflict reasons, most of the economic resources have been spent in defense purposes instead of being used in social areas, like education and health, highways and water supply, that’s one of the main reasons for the delay in the general development of Angola´s territory.

92

Processes of corruption in Portugal: results of an exploratory research

R. Faria1, C. da Agra2

1 School of Criminology, Faculty of Law, University of Porto, Portugal. 2 School of Criminology, Faculty of Law, University of Porto, Portugal.

Portugal lacks empirical research on corruption through a criminological approach [1]. The School of Criminology of Porto is trying to fill this empirical and theoretical gap by analysing literature proceeding from other social sciences (v.g. Law), collecting data and sketching the features of the usual actors and stages involved in this deviant behaviour, in our country. This exploratory research aims to enlighten the following items: (1) standings about the law, (2) black figures, (3) reasons and features of the complaints presented to the police; (4) the transition from the police system to the judicial; (4) dynamics of the trial; (5) type, incidence and features of the penal sanctions; (6) areas considered to be permeable to corruption; and (7) main causes for corruption practices. These items (except 6 and 7) follow the usual treatment given by the formal instances of control to the crime of corruption. For this purpose, we have endeavoured a documental analysis (Crime Statistics, reports from Transparency International, European Group of States against Corruption, and International Crime Victimization Survey) and conducted 5 semi-directive interviews with privileged informers. We were able to somehow understand the standings of the interviewees about the present laws of fight against corruption, as well as some interesting features on the complaints presented to the police, the handling of crimes of corruption by the judicial system and some of the main purposes of those who enact in these deviant behaviours. Besides, some results allow us to understand the crime of corruption as being a typical white-collar crime [2] or economic crime [3], eventually driven by rational choice [4-5].

References: [1] Faria, R. (2007), Corrupção: descrições e reflexões. Sobre a possibilidade de realização de uma abordagem criminológica ao fenómeno da corrupção em Portugal, Revista Portuguesa de Ciência Criminal, 17 (1), 107-146.

[2] Nelken, D. (2002), White-Collar Crime, in Maguire, Morgan and Reiner (Eds.), The Oxford Handbook of Criminology (3ª ed.), Oxford, Oxford University Press, pp. 844-877

[3] Ponsaers, P., and Ruggiero, V. (Eds.), (2002), La criminalité économique et financière en Europe, Paris, l'Harmattan.

[4] Cusson, M. (2006), Criminologia, Cruz Quebrada, Casa das Letras.

[5] Queloz, N., Borghi, M. and Cesoni, M.L., (2000), Processus de corruption en Suisse, Genève, Helbing & Lichtenhahn

93

Desistência do tratamento de enurese: avaliação dos motivos alegados e de variáveis associadas

Carolina Ribeiro B Sousa 1, Edwiges Ferreira de Mattos Silvares

1 Instituto de Psicologia da Univerasidade do S. Paulo, Brasil

Apresentação da Aluna: Aluna do Instituto de Psicologia e minha escolha por visitar a Universidade do Porto deve-se à abertura a área de Humanas e Humanidades demonstrada pela Instituição quanto ao meu próprio interesse em conhecer os laboratórios e a clínica- escola de lá. Pretendo trocar informações e, quem sabe, estabelecer um intercâmbio que ultrapasse esse encontro e se firme para futuras visitas, um trabalho multicêntrico e possibilidade de cursar doutorado. Acredito que esta possibilidade será muito rica para minha formação e fortalecer meu interesse em trabalhar na área de pesquisa e também me dedicar a carreira docente.

94

Popular culture and the formation of the working class

J. Aguiar1

1 Department of Sociology, Faculty of Humanities, University of Porto, Portugal.

Social class is sometimes considered as a mere economic or a political phenomenon. In our research we tried to relate social class with cultural variables. Most notably, our purpose is to show how a particular kind of cultural production – popular culture – affects social and political mobilization of the working class [1]. In this way, we focused our attention in the rural workers of the Left Margin of Guadiana River, in Alentejo. Basically, the research is a qualitative approach to the feelings and subjective apprehensions of those workers and how they interact with their political behaviour. Extensive interviews, oral testimonies, social photography and content analysis of poems and songs written by those social agents, are some of the techniques used to inquire our object of study. The application of twenty interviews – which culminated in the constitution of five life- stories – and the thorough examination of the content of more than three hundred popular poems and eighty-four songs, resumed a wide range of cultural elements in Alentejo’s workers. As the Bourdieu’s concept of habitus [2] illustrated, all the cultural heritage of these social agents contributed to the development of social and political practices based on values of solidarity and companionship. At the same time, the proper and autonomous production of their own cultural sociabilities induced mental and symbolic frameworks creating a sense of community and, which is further important, a real and effective community of individuals gathered by common values and beliefs. So, in order to understand some of the most relevant political interventions of Alentejo’s working class, such as the struggles and mass strikes against the dictatorship of Salazar or the process of land occupations in 1974 and 1975 [3], we cannot escape to integrate theoretically these most visible phenomena with the correlative cultural background that supports them. To summarize, cultural dimensions influence and interfere in political and socio-historical macro-processes. Without the dynamics of cultural self-identification of the collective of Alentejo’s workers – expressed in their popular culture – it would be hardly difficult for them to intervene in the political scene. References: [1] Thompson, E.P. (1991 [1963]), The making of the English working class 1780-1832, 4th edition, Penguin Books, London. [2] “Habitus is the matrix of dispositions of the objective structures socially incorporated in mental structures that human beings set off when they reflect about, apprehend and put practices in motion”. Bourdieu, P. (2002), Esboço de uma teoria da prática, Celta, Oeiras, pp. 167. [3] Murteira, A. (2004), Uma Revolução na Revolução: Reforma Agrária no Sul de Portugal, Campo das Letras and Câmara Municipal de Montemor-o-Novo, Porto and Montemor. pp.17-32. Barros, A. (1981), A Reforma Agrária em Portugal: das ocupações de terras à formação de novas unidades de produção. 2nd edition, Fundação Calouste Gulbenkian, Oeiras.

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A troubled legacy. Changing strategies of social transmission and patterns of identity production among industrial workers

Bruno Monteiro1

1 Sociologist. MA student (Sociology) in the Faculty of Humanities.

This paper`s aim to explore how the conjugation between structural economic and social transformations and the rearrangement of power relations in the shopfloor conducted to mutations in the strategies of identity production and transmission among different generations of working class agents. The author draws on a 4-months-long period working as a disqualified worker in a furniture factory and several long interviews to illustrate how this becomes an afflicted process and how new modalities to express the shopfloor everyday experience emerge. Worker`s collective memory is strongly supported in processes of oral and kinetic production, actualization and transmission that operates in the full spectre of working class social structuration places (like the shopfloor). Today, these institution rites happen to be alongside with alternative influences issued from school and mass culture. The youth avatars, as the consumerism and the urbanified references, are replacing the traditional workshop and virile culture, the dominant ethical behaviour (like austerity or oral-based mutual confidence) and the patterns of social relations with enterprise owners. The continuity of certain traits goes along with a different array of resistance tactics and personal expression in work and new arrangements of practices and representations about what must be work involvement. Notwithstanding being symbolically refused, worker`s social condition persists objectively re(as)sumed by the youth. Moreover, apparently emancipating acquisitions have bring, together with the maintenance of established patterns of mobilization to work (like bulimic behaviour in work searching for extra rent, resignation in the face of authoritarian hierarchies, perpetuation of informal economy), new dependencies and stronger subjections (like the progressive endebtement). Employment market transformations, along with new trends in hiring practices (expressing a bigger instability in contracts and in the economic market, which reflects also in higher unemployment rates), brings changes in the specific configurations of the household economy and, especially, in the familiar strategies of social reproduction. Particularly, the progressive technical depreciation of the older segments of the working force means also the refusal of the cultural definitions and interpretations that they carry with. The paulatine usury of workers minds and bodies, the obsolescence of that artisanal knowledge made of embodied schemes of doing and seeing the work and, finally, the disappearance of autonomy in work procedures, are gradually giving form to feelings of detachment among industrial worker. Thus promoting a sense of minority, this contributes to fossilize this category of social agents and induces the indignity (or impossibility) of social reproduction. In short, all this shape new trends in the collective and individual strategies activated to face a changing socio-economic environment and contributes to the complexification of the objective and symbolic transmission of the working class condition and identity.

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PARALLEL ORAL SESSIONS IV – POS.IV.3 THURSDAY, FEB. 21

IV.3 - LIFE & HEALTH III Laura Oliveira & Marcela Segundo Screening of Phenolic compounds with Bioactivity in Pieris brassicae L. Reared on Brassica rapa 27 var rapa L. using HPLC-DAD-MS/MS-ESI D. M. Pereira, A. Noites, F. Ferreres, P. Valentão, R. M. Seabra, P. B. Andrade Spatio-temporal characterization of the Endocannabinoid System enzymes-FAAH and COX-2 in the 69 fetoplacental unit A. Leite, B. Fonseca, N.A. Teixeira, G. Correia da Silva Determination of hypoxanthine in vitreous humor by a sequential injection analysis system and its 84 application in the estimation of postmortem interval Ana M. Santos, Ana I. Pereira, Marieta L. C. Passos, S. Frazão, Agostinho J.C. Santos, M. Lúcia M.

F. S. Saraiva, José L. F. C. Lima A new method to evaluate spontaneous pain in neuropathic pain models: a study in rats with spared 87 nerve injury A. Sequeira, I. Martins, S. Araújo, D. Lima, I. Tavares Comprehensibility of pictorial representations within health educational and instructional artifacts - a 167 case study E. Ferreira, S. Coutinho 212 Calcium uptake by rat hippocampal synaptosomes: regulation by different depolarizing agents A. Rocha, M.G.B. Lobo, and P. Correia-de-Sá 226 New Conditions for Greater Purification Yield of Phosphorylated Period Protein in Drosophila Christine M. Lomiguen, Isaac Edery

Screening of Phenolic compounds with Bioactivity in Pieris brassicae L. Reared on Brassica rapa var rapa L. using HPLC- DAD-MS/MS-ESI

D. M. Pereira1, A. Noites1 F. Ferreres2 P. Valentão1, R. M. Seabra1 and P. B. Andrade1

1REQUIMTE/ Department of Pharmacognosy, Faculty of Pharmacy, University of Oporto, Portugal email: [email protected] 2Research Group on Quality, Safety and Bioactivity of Plant Foods, Department of Food Science and Technology, CEBAS (CSIC), Murcia, Spain

In this research we proceeded to the study of Pieris brassicae, a frequent pest of some species, as a source of compounds with interest for health. The phenolic profiles of P. brassicae at different development stages (larvae, exuviae and butterfly), its excrements and its host plant, Brassica rapa var. rapa L., were determined by HPLC-DAD-MS/MS- ESI (Fig.1). Twenty five acylated and nonacylated flavonoid glycosides, as well as ferulic and sinapic acids, were identified in host plant, from which only twelve compounds were found in the excrements. In addition, the excrements showed the presence of sulphate flavonoids and other flavonoid glycosides, not detected in the leaves of the host plant. In the larvae kept without food for twelve hours, only three compounds common to the plant material and two others also present in the excrements were characterized.

Brassica rapa var rapa Pieris brassicae

Fig. 1. Detection at 330 nm. (1) kaempferol-3-O-(methoxycaffeoyl)sophoroside-7-O-glucoside; (2) quercetin-3-O- (sinapoyl)sophoroside-7-O-glucoside; (3) quercetin-3-O-(feruloyl)sophoroside-7-O-glucoside; (4) quercetin-3-O- (p-coumaroyl)sophoroside-7-O-glucoside; (5) kaempferol-3-O-(sinapoyl)sophoroside-7-O-glucoside; (6) kaempferol-3-O-(feruloyl)sophoroside-7-O-glucoside; (7) kaempferol-3-O-(p-coumaroyl)sophoroside-7-O- glucoside; (8) p-coumaric acid; (9) kaempferol-3,7-di-O-glucoside; (10) isorhamnetin-3,7-di-O-glucoside; (11) ferulic acid; (12) sinapic acid; (13) kaempferol-3-O-(feruloyl)sophoroside-7-O-glucoside (isomere); (14) kaempferol-3-O-(methoxycaffeoyl)sophoroside; (15) kaempferol-3-O-(caffeoyl)sophoroside; (16) kaempferol-3- O-sophoroside; (17) kaempferol-3-O-(feruloyl)sophoroside; (18) kaempferol-3-O-glucoside; (19) isorhamnetin-3- O-glucoside.(20) quercetin-3-O-sophoroside; (21) kaempferol-3-O-(p-coumaroyl)sophoroside (isomere).

Antioxidant potential was studied against DPPH. radical and a reactive oxygen species (superoxide radical). The three samples showed an effective activity against both radicals in a concentration-dependant manner, with larvae displaying the strongest capacity against DPPH., while excrements were the most effective superoxide radical scavenger. So, the matrices studied herein, may constitute a promising source of bioactive complex compounds, hard to be isolated or synthesized in the laboratory due to its high structural complexity.

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Spatio-temporal characterization of the Endocannabinoid System enzymes-FAAH and COX-2 in the fetoplacental unit

A. Leite 1, B. Fonseca 1,2, N.A. Teixeira 1,2 and G. Correia da Silva 1,2

1Laboratory of Biochemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 IBMC- Institute of Molecular and Cellular Biology, University of Porto, Portugal.

The comprehension of the mechanisms underlying the effects caused by the consumption of Cannabis sativa was essential for the discovery of an endogenous complex signalling system – the endocannabinoid system. Anandamide (AEA), the main endocannabinoid (EC), which mimics the action of exogenous cannabinoids, was the first ligand to be discovered and is able to bind to the cannabinoid receptors CB1, CB2 and to the vanilloid receptor VR1. AEA is locally released from its phospholipid precursor by a N- acylphosphatidylethanolamine (NAPE)-specific phospholipase D (NAPE-PLD) and the levels of AEA are regulated by its inactivation namely via a fatty acid amide hydrolase (FAAH) and a cyclooxygenase (COX-2). Previous studies have demonstrated that the components of this system are important for the maintenance of the low endocannabinoid levels necessary for the correct blastocyst implantation [1]. However, little is known on the physiological significance of AEA during fetoplacental development. Therefore, using the rat as the model for haemochorial placentation, we examined the spatiotemporal expression patterns of FAAH and COX-2 by immunohistochemistry and Western blot on days 8, 10, 12, 14, 16 and 19 of the gestational period. The results demonstrate the presence of FAAH and COX-2 in maternal tissues, namely in decidual cells and in NK cells, during the process of decidual remodelling and placental development, decreasing its expression throughout pregnancy. Therefore, the expression of the enzymes responsible for the inactivation of endocannabinoids associated with the presence of CB and VR1 receptors in decidual cells suggests a relevant role for this system during placental development. The study of the expression of the enzymes and receptors involved in the regulation of this system becomes clinically relevant, since the EC present dual functionality, that is, when expressed in low levels they perform their physiological regulatory functions, whereas its deregulation and inherent high levels of these ligands are associated with spontaneous pregnancy loss. At this point it is mandatory to remember that the consumption of Cannabis (which uses the same metabolism such as EC) during pregnancy may alter de enzymatic expression in addition to the endocannabinoid levels, consequently, originating a series of pregnancy dysfunctions as a result of abnormal and non-functional placental development. Therefore, the endocannabinoid system may be a potential important target concerning human infertility.

References: [1] Wang, H., Xie, H., Sun, X., Kingsley, J.P., Lawrence, J., Cravatt, B.F. and Dey, S.K. (2007), Differential regulation of endocannabinoid synthesis and degradation in the uterus during embryo implantation, Prostaglandins & Other Lipid Mediators, 83, 62–74.

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Determination of hypoxanthine in vitreous humor by a sequential injection analysis system and its application in the estimation of postmortem interval

Ana M. Santos1, Ana I. Pereira1, Marieta L. C. Passos1, S. Frazão2, Agostinho J.C. Santos2, M. Lúcia M. F. S. Saraiva1 and José L. F. C. Lima1

1 REQUIMTE, Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Portugal. 2 Serviço de Medicina Legal, Faculdade de Medicina, Universidade do Porto, Portugal. Serviço de Patologia Forense, Instituto Nacional de Medicina Legal, I.P. – Delegação Norte.

The estimation of the postmortem interval (PMI), i. e., the estimation of the time since death, is crucial to the practice of certain forensic cases. Many methods have been used to estimate the PMI, and the determination of hypoxanthine is one of them. Hypoxanthine is a degradation product of adenosine. Increased concentrations were reported in cases of hypoxia as well as with prolonged PMI. Hypoxanthine is recommended as an indicator of prolonged (cerebral) hypoxia, for example in victims of sudden infant death as well as a new biochemical method for estimation of postmortem time [1]. In this work it is proposed to estimate the PMI on the basis of postmortem hypoxanthine concentrations in vitreous humor. The hypoxanthine concentration is determined by the enzymatic oxidation of the hypoxanthine to uric acid. The enzyme xanthine oxidase catalyzes the oxidation of hypoxanthine in two steps producing xanthine in the first and uric acid in the second, which is detected spectrophotometrically at 290 nm. This reaction is applied in a sequential injection analysis (SIA) system [2] where it was incorporated a column with the xanthine oxidase immobilized in activated glass beads. Physical and chemical parameters such as pH, concentration of reagents, temperature, and sample volume were studied in order to attain the best analytical performance and the lower reagent consumption. The specificity of this reaction associated with the use of an automatic SIA system constitutes a robust, versatile, and simple alternative, adapted to the actual demands of sample and reagents economy and reduced residue production.

References: [1] Madea, B., Kaferstein, H., Hermann, N. and Stricht, G. (1994), Hypoxanthine in vitreous-humor and cerebrospinal-fluid – a maker of postmortem interval and prolonged (vital) hypoxia – remarks also on hypoxanthine in sidsl, Forensic Sci Int., 65 (1), 19-31. [2] Ruzicka, J. and Marshal G. D. (1990), Sequential injection: a new concept for chemical sensors, process analysis and laboratory assays, Analytica Chimica Acta, 237, 329-343.

Acknowledgements: Marieta L. C. Passos thanks Fundação para a Ciência e a Tecnologia and FSE (III Quadro Comunitário) for the Ph.D. grant (SFRH / BD / 22752 / 2005). Authors thanks Reitoria da Universidade do Porto the Project Investigação Científica na Pré-Graduação (2007).

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A new method to evaluate spontaneous pain in neuropathic pain models: a study in rats with spared nerve injury

A. Sequeira1, 2, I. Martins1, 2, S. Araújo1, 2, D. Lima2 and I. Tavares1

1 Institute of Histology and Embryology, Faculty of Medicine, University of Porto and IBMC, Portugal. 2 Laboratory of Molecular Cell Biology, Faculty of Medicine, University of Porto and IBMC, Portugal.

Patients with neuropathic pain report spontaneous pain along with evoked allodynia and hyperalgesia. However, only evoked reactions have been used to study the magnitude of neuropathic pain, since models to evaluate spontaneous pain are scarce. Based on the relevance of the spared nerve injury (SNI) model in long-term studies of neuropathic pain, we performed an overtime evaluation of spontaneous pain in SNI rats in comparison with evoked pain. Male Wistar rats were submitted to SNI induction (n=8) or sham-operation (n=6). Evoked and spontaneous pain reactions were evaluated immediately after surgery and at 1, 3 and 6 months later. Evoked pain was studied using the von Frey, acetone and pin-prick tests. In spontaneous pain, animal behavior was evaluated during 10 minutes using Etholog2.2® and two types of reactions were analyzed: pain-traducing and physiological behaviors. Pain- traducing behaviors included several reactions directed to the hindpaw ipsilateral to the injured nerve, such as freezing and shaking, scratching, lifting and licking. Physiological reactions included exploring, rearing, washing and all routine animal behaviors. Before SNI induction and in sham-operated rats, evoked pain behaviors were reduced and spontaneous pain reactions were absent. In SNI animals, the increase in evoked pain behavior was more evident one month after surgery. These animals also presented a decrease in physiological behavior. Spontaneous pain-traducing actions remained elevated during all analyzed time points. Based on the relation between evoked and spontaneous pain detected in SNI rats, the present study shows that the method of spontaneous pain analysis is a reliable protocol to fully behaviorally access animals with chronic pain.

KEY WORDS: neuropathic pain, spontaneous pain, evoked pain, allodynia, hyperalgesia.

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Comprehensibility of pictorial representations within health educational and instructional artifacts – a case study

E. Ferreira 1 and S. Coutinho2

1 Department of Design, Faculty of Design, Universidade Federal de Pernambuco, studying at present at Department of Design, Faculty of Belas Artes, University of Porto, Portugal. 2 Department of Design, Faculty of Design, Universidade Federal de Pernambuco).

This paper presents a case study that investigates the comprehensibility of illustrations presented in educational and instructional artifacts from the health area, according to the model proposed by Goldsmith [1] and an experimental study. We present the foundations of the mentioned study, the applied methodology and indications of results, concluding with the identification of failures in the intended pictorial communication. The artifacts used for this study were collected on April and May of 2006 at Recife, Brazil. From the initial sample (32 artifacts), we’ve categorized and selected the most representative material in terms of illustrations, to proceed with the work. The subject of the selected material is the schistosome disease, very common in Brazil. The experimental study considered two groups: a control group (presented to isolated images) and an experimental group (with images viewed on the original artifact). As common procedure, the participants of both groups were asked to talk about all their perceptions of the pictures. The researchers made use of a tape recorder and notes; the experiment took place on October 06th, 2006, at Hospital das Clínicas da UFPE. Considering those perceptions, the analytical approach of this study made use of Goldsmith´s model, where the author purposes that there must be considered visual factors (unity, location, emphasis, parallel text) and semiotic levels (syntactic, semantic, pragmatic) in a related way, providing a structure on which an illustration can be evaluated in terms of accessibility to information. According to Goldsmith, a message should properly fit the signs (syntactic matter) in a way that the public understands its meanings (semantic) and there is affinity between the purpose of the message and the context of the observer (pragmatics). The author suggests that her model can be used to determine the reasons why the comprehensibility of the illustrations didn’t occurred properly. As a result of this study, we could identify communicational failures on each of the purposed levels (syntactic, semantic, pragmatic), considering also the lack of familiarity of the public with the pictorial representations exposed. Actually, the information was mostly misunderstood or partially comprehended. With this study, the experiment and the analytical model applied, we could contextualize and understand better some of the failures; as further results, we can then purpose new and more accurate solutions.

References: [1] Golsmith, E. (1984), Research into illustration: an approach and a review. Cambridge: Cambridge University Press.

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Calcium uptake by rat hippocampal synaptosomes: regulation by endogenous adenosine and different depolarizing agents

A. Rocha, M.G.B. Lobo, and P. Correia-de-Sá

Laboratório de Farmacologia e Neurobiologia, UMIB, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Portugal.

Adenosine is an endogenous modulator of the CNS refraining neuronal excitability (e.g. [1]). Because endogenous levels of adenosine rise markedly during seizure activity, this nucleoside has been proposed as an “endogenous anticonvulsant”. The anticonvulsant effect of adenosine may be 2+ due to A receptors activation [2] leading to suppression of voltage-dependent Ca entry [3]. 1 Animal models and human tissue (from epileptics submitted to neurosurgery) exhibit similar mechanisms underlying epileptic disorders [4]. Synaptosomes – isolated nerve endings that 2+ maintain physiological properties such as the ability to release neurotransmitters in a Ca - dependent manner - are throughly used to study pathogenesis of epilepsy. Veratridine (VT, a + + + selective Na opener), 4-aminopyridine (4-AP, a nonselective K channel blocker) and high K concentrations are the most common strategies to depolarise synaptosomal plasma membrane as + they allow to differentiate between presynaptic Na channel-mediated responses; these compounds 2+ increase intracellular Ca accumulation in the rat hippocampus as well as in the human brain. + Nevertheless, we recently showed that the choice of depolarising agent (VT vs high [K ]) may be 2+ crucial for evidencing paradoxical effects of drugs regulating Ca influx into hippocampal synaptosomes, such as nifedipine [5]. Therefore, we investigated whether the same occurred using 4-AP as compared with VT. We also tested the modulatory role of endogenous adenosine on 2+ neuronal Ca uptake by incubating hippocampal synaptosomes with adenosine deaminase (ADA), which inactivates adenosine into inosine. Synaptosomes were prepared from rat hippocampal homogenates [5]. Samples (50 μl) of synaptosomal suspension were pre-incubated with test drugs for 10 min, at 37 ºC. Synaptosomal 45 loading was performed during 3 min with CaCl (0.32 μCi/μmol) in the presence of the 2 depolarizing agent. Adding ice-cold Tris-EGTA solution followed by filtration under negative 45 pressure through GF/C glass fibre filters, terminated Ca uptake. The radioactivity associated with the synaptosomes was determined by liquid scintillation spectrometry. -1 45 Inactivation of endogenous adenosine with ADA (0.5 UmL ) caused a global increase on Ca uptake into the rat hippocampal synaptosomes, indicating that there is a significant adenosine 2+ inhibitory tonus regulating depolarisation-induced Ca influx into nerve terminals. VT (5 μM) 45 increased Ca uptake into hippocampal synaptosomes (67±9 %, n=4). 4-AP (10 mM) increased 45 Ca uptake to a lesser extent (23±4%, n=4). Co-application of VT (5 μM) together with 4-AP (10 45 mM) increased Ca uptake (138±15 % (n=4) above the control level. This higher-than-additive effect, contrasts with the additive result obtained upon applying VT (1-10 μM) together with high + [K ] (15-30 mM) [5], indicating the existence of synergism between VT and 4-AP.

[1] Cunha, R. Neurochem. Int. 38, 107-125, 2001 [2] Zhang, G., Franklin, P.H. & Murray, Eur. J. Pharmacol.,255, 239-243,1994 [3] Wu, L.G. & Sagau, P, Neuron, 12, 1139-1148, 1997 [4] Avoli,M., Louvel,J., Pumain,R. & Kohling,R., Prog. Neurobiol., 22, 166-200, 2005 [5] Costa, J. Lobo, M.G.B. & Correia-de-Sá., Eur. J. Pharmacol., 544, 39-48, 2006

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New Conditions for Greater Purification Yield of Phosphorylated Period Protein in Drosophila

Christine M. Lomiguen1, Isaac Edery2

1Biomedical Engineering, Rutgers The State University of New Jersey 2Molecular Biology and Biochemistry, Rutgers The State University of New Jersey

A conserved feature of animal clocks is that period (per) proteins undergo daily oscillations in levels and phosphorylation states regulated by doubletime (dbt in Drosophila) [Casein kinase I (CK1 in mammals)]. The phosphorylation of PER has been shown to affect its biochemical function and metabolism, including subcellular localization, protein-protein interactions, as well as targeted degradation through the proteasome. There is evidence that a mutation (S662G) in human PER2 leads to phosphorylation defects and is associated with familial advanced sleep phase syndrome (FASPS). The identification of PER phosphorylation sites is critical to understanding the biochemical function of PER. To map these phosphorylation sites, a large amount of PER must be purified and subjected to mass spectrometry. For this research, we attempted to determine 1) the conditions for obtaining hyperphosphorylated PER proteins in Drosophila S2 cell culture; 2) the lysis buffer that should be used for the purification for His-tagged PER. Our results show that in order to purify large quantities of hyperphosphorylated PER isoforms, we should use stable cell lines expressing both PER and DBT. The transient transfection of the Sb(∆F) plasmid into the PER+DBT stable cell line is not likely going to improve the yield for hyperphosphorylated PER isoforms. On the other hand, we show that MG132 has a positive effect in increasing hyperphosphorylated PER.

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POSTERS WEDNESDAY & THURSDAY, FEB. 20&21

28 HPLC-DAD-MS/MS analysis of phenolics in vitro shoots of Brassica oleracea L. var. costata DC M. Taveira, C. Sousa, F. Ferreres, D. M. Pereira, Paula B. Andrade, R. M. Seabra, Pedro

Marques, Patrícia Valentão 39 Thermodynamic and Theoretical Study of Diphenylpyridines Marisa A. A. Rocha, Carlos F. R. A. C. Lima, Lígia R. Gomes, Luís M. N. B. F. Santos 40 Form and Structure on Eduardo Souto de Moura's architecture. B. Moreira, C. Machado 46 Possible correlation between morbid obesity and inflammation: a peripheral leukocyte study A. Meneses, I. Maio, D. Pestana, P. Freitas, R. Monteiro, I. Azevedo, C. Calhau, R. Soares In vitro and in vivo studies of the expression of carbohydrates in a canine mammary carcinoma cell 47 line J. Gomes, C. Lopes, E. Hellmén, C. Reis, F. Gärtner 49 Determination of Cadmium Content in Sardine pilchardus Ana C. Silva, S. Morais, A. Novo, C. Luz, E. Pinho, S. Conceição, C. Delerue-Matos, M.B.P.P.

Oliveira Optimization Study for Arsenic Quantification in Fish Using Microwave-assisted Digestion and 50 Graphite Furnace Atomic Absorption Spectrometry Ana C. Silva, S. Morais, A. Novo, C. Luz, E. Pinho, S. Conceição, C. Delerue-Matos, M.B.P.P.

Oliveira Optimization of an enzymatic and chromatographic method for inorganic pyrophosphate based on 51 firefly luciferase Simone M. Marques, Joaquim C.G. Esteves da Silva 53 Survey of the effects of the antibiotic minocycline on freshwater cyanobacteria J.Pereira, M. S. Baptista, T. Stoichev, B. Heggie, V.M. Vasconcelos, M.T.S.D. Vasconcelos 54 Low-cost materials as adsorbents for Textile Wastewater Treatment S. Santos, R. Deucher, R. Boaventura 55 Determination of physical parameters in pharmaceutical powders by near infrared spectroscopy (NIR) Ana Cruz, Sandra Soares, Mafalda C. Sarraguça, Paulo C. Costa, Helena R. Amaral, João A.

Lopes An expeditious method for determination of residues of organochlorine pesticides in sediments and 59 its application to Portuguese estuarine areas P. N. R. Rodrigues, P. N. Carvalho, F. Alves, R. Evangelista, M. C. P.Basto, M. T. S.

D.Vasconcelos 61 Kinetics of Inhibition of Firefly Luciferase by Oxyluciferin and Dehydroluciferyl-adenylate César Ribeiro, Joaquim C.G. Esteves da Silva 64 Effect of pyrophosphate on firefly luciferase bioluminescence Filipe Peralta, Joaquim C.G. Esteves da Silva, Rui Fontes 66 Molecular Energetics of Hydroxybenzaldehyde Isomers Mariana Vidinha, Manuel J. S. Monte, Maria D. M. C. Ribeiro da Silva 67 Touristic Topographies : transversality and authenticity in an Portuguese context João Nuno Pinto Marques 68 Sensor for amperometric determination of ammonia in seawater C. Peixe, C.M. Pereira, F. Silva, M-T 70 Copper(II)/Ciprofloxacin complexes: Synthesis and Solution Studies C. Queirós, I. Sousa, E. Pereira, P.Gameiro THERMOCHEMICAL STUDIES OF TWO CYCLIC UREAS: IMIDAZOLINONE AND N,N'- 71 TRIMETHYLENEUREA V. Freitas, M. Roux, J. Elguero, M. Ribeiro da Silva 72 Feeling Architecture. The importance of 'the other' senses in the experience of space. S. Silva Natária 73 Deep Eutectic solvents: A new kind of Ionic Liquids R. Costa, F. Silva, C.M. Pereira Authenticity evaluation and molecular characterization of Portuguese varieties of Juglans regia using 75 microsatellite markers. E.M. Cabral, M.A. Faria, M.B.P.P. Oliveira

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76 UV PROTECTION OF GEOTEXTILES BY CHIMASSORB 944 Inês Valente, Paulo Almeida, José Ricardo Carneiro, Maria de Lurdes Lopes 77 Computational Analisys of Cynara cardunculus L. Cardosins Pomoters Alberto M. Pessoa, Jorge Teixeira, José Pissarra 78 Hydrogen Peroxide Reversible Luminescence Sensors Célia Rocha, Joaquim C.G. Esteves da Silva 79 Chromium remediation using zero valent iron - batch studies G. Carvalho, Aurora Silva, Cristina Delerue-Matos, A. Vega y Fuente, António Fiúza 80 Development of a multicommutated flow system for chemiluminometric quantification of gentamicin L. Santos, A. Araújo, A. Fachini, M. Montenegro, B.F. Reis 81 Potentiometric determination of gentamicin in pharmaceutical formulations Martina Koci, Célia Amorim, Alberto Araújo, Maria da Conceição Montenegro, Marie Pospíšilova,

Petr Solich 82 Influence of organic pollutants on phytoremediation of copper by a salt marsh plant A. C. M. Dias, C. M. R. Almeida, A. P. Mucha, M. T. S. D.Vasconcelos DFT STUDY ON THE ABILITY OF CALIX[2]FURANO[2]PYRROLE TO FORM HOST-GUEST 88 COMPLEXES WITH DIFFERENT IONS C.A. Teixeira dos Santos, A.L. Magalhães 91 Fiber optic pH sensors based on CdTe quantum dots Helena Gonçalves, Conceição Mendonça, Joaquim C.G. Esteves da Silva, César Maule, Pedro

Jorge 92 Steady state and lifetime chemical sensors based on Ruthenium complexes Helena Gonçalves, Joaquim C.G. Esteves da Silva, César Maule, Pedro Jorge Molecular characterization of the Portuguese patients with Mucopolysaccharidosis IIIC: two novel 94 mutations in the HGSNAT gene Coutinho MF, Lacerda L, Prata MJ, Ribeiro H, Lopes L, Ferreira C, Alves S 95 Scopoletin, a natural coumarin isolated from Agrostistachys gaudichaudii (Euphorbiaceae) C. Silva, A.E. Oliveira, A.P. Almeida, H. Cidade, A. Kijjoa, M.S.J. Nascimento 5-(Hydroxymethyl)-2-furfural, a natural compound isolated from Strophioblachia fimbricalyx Boerl. 96 (Euphorbiaceae) K.C. Marques, A.E. Oliveira, A.P. Almeida, H. Cidade, A. Kijjoa, M.S.J. Nascimento New approaches for estimating the postmortem interval based on vitreous humor trace element 97 levels H. Andrade, A. Almeida, S. Frazão, T. Magalhães, A. Santos New approaches to the diagnostic of death by drowning based on trace element levels in blood of 98 the cardiac cavities F. Pereira, T. Magalhães, A. Almeida, A. Santos, B. Santos Firing distance estimation through the analysis of the gunshot residues deposit pattern around de 99 bullet entrance hole J. Tuna, A. Santos, A. Almeida, T. Magalhães In vitro assessment of whey edible coats with antimicrobial features against differents microbial 103 strains A. Santos, M. Leão, O. Ramos, M. Pintado, X. Malcata. Synthesis, structure elucidation and evaluation of xanthone derivatives for dual activity: antitumor 106 activity and P-glycoprotein inhibition J. Lima, A. Paiva, E. Sousa, M. Pinto, M. S. J. Nascimento, M. H. Vasconcelos 107 Synthesis of xanthone derivates for in vitro and in vivo biological activity studies J. Siroka, E. Sousa, M. Pinto 109 Ultrastructural characterisation of Pirellula sp. OJF20 R.Abreu, F.Viana, J. Torres, J. Bondoso, A. Lobo-da-Cunha, O. M. Lage 110 Mesoporous Carbons: Synthesis and Functionalization C.A. Orge, J.P.S. Sousa, F. Gonçalves, C. Freire, M.F.R. Pereira, J.J.M. Órfão 112 Physical exercise and stress hormones M. Moz, S. Baptista, O. Cardoso, M. Mendanha, A. Ascensão, J. Magalhães, L. Ribeiro 113 Single-standard calibration applied to SIA analytical system with potentiometric detection Leonel G. C. Soares, Rui A.S. Lapa

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PHOTOCATALYTIC DEGRADATION OF CIPROFLOXACIN ANTIBIOTIC IN TiO2 AQUEOUS 115 SUSPENSION F. G. Azevedo, M. Domingos, C. G. Silva, J. L. Faria Anti-Angiogenic Effects of Ranibizumab and Bevacizumab in Age-Related Macular Degeneration: 123 Effects on Human Endothelial Cells M. Ferreira-Pinto, T. Taveira-Gomes, A Carneiro, M Falcão, I. Azevedo, F. Falcão-Reis, R.

Soares 125 Geometry and Space Conception the importance of the processes of representation to Architecture Bruno Russo, João Pedro Xavier 127 Wet Peroxide Oxidation and Wet Oxidation of Nitrophenols in Aqueous Streams S. Martins, A. Ribeiro, A.M.T. Silva, P. Araújo, J.L. Figueiredo, J.L. Faria 129 Formation of heterocyclic aromatic amines in charcoal grilled sardines M. Costa, A. Melo, C. Petisca, I.M.P.L.V.O. Ferreira, O. Pinho Influence of beer antioxidative properties on the formation of the heterocyclic amine PhIP in grilled 130 beef A. Melo, M. Costa, R. Eça, I.M.P.L.V.O. Ferreira, O. Pinho 131 THE STUDY OF DE FÂNZERES-GONDOMAR GRANITES: A MULTIDISCIPLINARY APROACH M. Alves Educational Sciences and Medical Education: An internship in FCM-UNL Department of Medical 135 Education R. Encarnação 136 Quartz Crystal Microbalance: A Useful Tool As Sensor Of Thin Films J. Borges, C. M. Pereira, F. Silva 137 João Andresen's Fifties Houses: Portuguese Domestic Architecture Up-to-Date Sergio Dias da Silva 138 Synthesis of xanthone derivatives by microwave-assisted methods P. Vrbata, M. Pinto, R. Castanheiro, S. Cravo 139 Location and Ownership of the Natural Resources in Iraq C. Silva, H. Azevedo Synthesis, Structural Elucidation and Biological Activities of Chiral 1-Amino-2-Propanol Xanthone 140 Derivatives I. Teixeira, M. Pinto, C. Fernandes 141 Chiral Alaninol Xanthone Derivatives: Synthesis, Structural Elucidation and Biological Activities A. Pinto, C. Fernandes, M. Pinto, A. Pozzi, B. Matos, L. Oliveira, P. Correia-de-Sá 143 Primary Hyperparathyroidism: the unknown world of sporadic cases and their genetic alterations. Alvelos M., Barbosa E., Soares P. Flow cytometric detection of P-gp mediated drug efflux: a method for the evaluation of the activity of 144 P-gp inhibitors A. Palmeira, R. Lima, H. Seca, M.H. Vasconcelos, E. Sousa, M. Pinto 145 Effects of etoposide, doxorubicin and cytarabine in Burkitt Lymphoma cell lines R. Lima, H. Seca, M. I. Castro, S. Brás, P.Soares, M.S. Nascimento, M. H. Vasconcelos 146 Automatic FIA method for determination of iron in environmental samples E. P. Freitas, F. A. Dias, H. M. Oliveira, M. A. Segundo, S. Reis, J. L. F. C. Lima 147 Characterization of exotic wood species for finishing with water-borne varnishes D. Martins , V. Lopes, F. Oliveira, A. Mendes, L. Carvalho 154 Microbiological study of the interaction of Outer Membrane Proteins with antibiotics M. Garrido, P. Gameiro, P.J. Eaton, M. Feio Interactions of sulindac and its metabolites with phospholipid membranes: an explanation for the 156 peroxidation protective effect of the bioactive metabolite F. Santos, L. Teixeira, M. Lúcio, J. L. F. C. Lima, S. Reis 158 Evaluation of primer coatings on steel subtracts by a low-cost, fast electrochemical technique V. Gonçalves, A. Mendes, J. Machado, F. Oliveira, J. Nogueira, H. Aguilar Ribeiro 161 The Japanese Influence in Modern Western Architecture -an introduction- Miki Itabashi 163 Purinergic signalling in a model of overactive urinary bladder Nuno Silva, Miguel Faria, Vítor Cavadas, Paulo Correia-de-Sá

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166 Conduction block in isolated rat sciatic nerve by chiral aminoalkanolic derivatives of xanthones Bernardo Matos, Angelo Pozzi, Ana Pinto, Inês Teixeira, Emidio Fernandes, David Moreira, Carla

Fernandes, Laura Oliveira, Madalena Pinto, Paulo Correia-de-Sá 169 Osteoclastic resorption of calcium phosphate based bone substitutes: in vitro studies C.A. Teixeira, J. Costa-Rodrigues, P.S. Gomes, M.H. Fernandes Monitoring particulate matter using magnetic properties of tree leaves in the Porto urban area and 170 Cinfães area. H. Corrêa-Ribeiro Adenosine regulates its own release from myenteric neurons via A2A receptors coupled to adenylate 173 cyclase / cAMP pathway J. Duarte, R. Dias, M. Duarte-Araújo, T. Magalhães-Cardoso, P. Correia-de-Sá 175 Education and Poverty: an empowerment approach A. Oliveira 177 Synthesis of vesiculated polyester particles and its incorporation in paints A. Dias, A. Mendes, F. Oliveira, J. Moniz, F.D. Magalhães 178 Characterization of Urea-Formaldehyde Resins F. Silva, J. Ferra, L. Carvalho, J.P. Liberal, F. D. Magalhães Characterisation of the Planctomycetes Pirellula sp. OJF20 and Pirellula sp. OJF27 isolated from the 179 surface of macroalgae F. Viana, J. Torres, R. Abreu, J. Bondoso, O. Lage 180 Investigation of the Interactions between Phospholipids and Gold Nanoparticles at Air/water Interface C. Carreira, F. Silva, C. Pereira Development and validation of a new multiresidue method for the determination of 17 polychlorinated 181 dibenzodioxins (dioxins) and polychlorinated dibenzofurans (furans) in environmental matrices by SPME-GC-MS A.Neves, A.D. Guimarães, M.F. Alpendurada 184 ADENOSINE REGULATES DIFFERENTIATION OF HUMAN OSTEOBLAST CELLS IN CULTURE A. Barbosa, M.A. Costa, T. Magalhães-Cardoso, A. Teixeira, R. Freitas, J.M. Neves, P. Correia-

de-Sá 185 The Pombais (pigeon houses) in the northeast Transmontano (northeast of Portugal) Mafalda Matias, Teresa Fonseca 187 Pravastatin Quantification using Square-Wave Voltammetry M. Neves, S. Pereira, H. Nouws, C. Delerue-Matos 191 Microbial community distribution and dynamics in a shallow raceway-recirculating mariculture system A. Matos, M.-T. Borges, P.M.L. Castro 194 Information Systems about Materials for Architecture F. Figueiredo Osteogenic potential of adipose tissue-derived mesenchymal stem cells. Effect of medium 196 composition and cell passage C. Bessa Pereira, R. Almeida Palmas, M.H. Fernandes 197 Potential of Equisetum arvense hydromethanolic extracts as osteoblastic growth promoters C. Bessa Pereira, L. Vieira, M.A. Lopes, M.H. Fernandes Comparison of the in vivo bone forming activity of osteogenic-induced mesenchymal stem cells 198 derived from adipose tissue and bone marrow T. Ribeiro, C. Bessa Pereira, R. Almeida Palmas, P. Gomes, M.H. Fernandes 199 Taxonomy from rural diffuse to urban industrial diffuse André Chaves 202 Tetracyclines modulation of the osteogenic differentiation - in vitro evaluation I. Côrte-Real, A. L. Marinho, P. S. Gomes, J. Rodrigues, M. H. Fernandes 204 Study and design of precast reinforced concrete box-culverts under high embankments Daniel Pinto 207 Properties of nanostructured materials C. P. Arieira, F. Silva, C. M. Pereira Influence of egg yolk and aromatizing compounds on the volatile profile and on sensory 208 characteristics of bakery cream B. Ramos, O. Pinho, I.M.P.L.V.O. Ferreira

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209 Evaluation of genetic and chemical diversity in Portuguese populations of Phaseolus vulgaris L. R. Coelho, M. A. Faria, A. Barata, M.B.P.P. Oliveira, E. Nunes 210 The metamorphosis of Aleixo towers A. Lima 214 Modulation of cytochrome P450 and oxidative stress by rooibos (Aspalathus linearis) infusion intake D. Teixeira, A. Marinho, C. Meneses, I. Ferreira, J. Sampaio, M. A. Lima, V. Mendes, A. Faria, R.

Monteiro, C. Calhau 216 Screening for novel CDH1 inactivating mechanisms in Familial Gastric Cancer P. Inácio, H. Pinheiro, J. Carvalho, S. Sousa, R. Seruca, C. Oliveira 217 The influence of interpersonal similarity in social categorization D. Teixeira, F. Gonçalves, I. Lourenço, S. Duarte Genetic characterization by SSR of six Olea europaea regional varieties for authenticity evaluation of 220 olive oils J. Lopes, R. Coelho, M. A. Faria, E. Nunes, M.B.P.P Oliveira 228 Alternative User Interfaces for Game Control Sarah Imboden Permeability Characteristics of High Performance Concrete (HPC) Containing Varying 221 Concentrations of Hycrete Tested and Compared by Method of Rapid Chloride Permeability Test and the Non-Steady-State Migration Test (nordtest method NT Build 492) Michael Boxer, Hani Nassif Crises e Reformas: o colonialismo ilustrado na Capitania de São Paulo e os esforços de 222 modernização do Governo Morgado de Mateus (1765-1775) Joana Salém Vasconcelos, Vera Lúcia Amaal Ferlini 223 Desistência do tratamento de enurese: avaliação dos motives alegados e de variáveis associadas Carolina Ribeiro Bezerra de Sousa, Edwiges Ferreira de Mattos Silvares 224 Mario Pedrosa e sua atuação crítica na Presidência da ABCA Gabriela Borges Abraços 226 New Conditions for Greater Purification Yield of Phosphorylated Period Protein in Drosophila Christine M. Lomiguen, Isaac Edery 227 Intracellular Gene Regulation via Engineered Nanolipoblockers for Inhibition of Cholesterol Uptake Ekta Patel, Prabhas Moghe

110

HPLC-DAD-MS/MS analysis of phenolics in vitro shoots of Brassica oleracea L. var. costata DC

M. Taveira1, C. Sousa1, F. Ferreres2, D. M. Pereira1, Paula B. Andrade1, R. M. Seabra1, Pedro Marques1, Patrícia Valentão1

1 REQUIMTE/Department of Pharmacognosy, Faculty of Pharmacy, University of Oporto, Oporto, Portugal. [email protected] 2 Research Group on Quality Safety and Bioactivity of Plant Foods, Department of Food Science and Technology, CEBAS (CSIC), P.O Box 164, 30100 Campus University of Espinardo, Murcia, Spain.

Brassica oleracea is native of the Mediterranean region and southwestern Europe, with a great importance in human nutrition. Brassica species, including B. oleracea, are reported to possess antioxidant activity, which is partially attributed to their phenolic compounds. In vitro culture of plants is a method of biomass production, which enables production of large number of species in a short period of time and space. Different conditions and supplements used in medium constitution affect plant viability, growing and production of specific metabolites. For the first time, phenolic compounds produced by in vitro shoots of B. oleracea L. var. costata DC were determined by HPLC-DAD-MS/MS (Fig 1). Thirty seven compounds were characterized, which included chlorogenic acids, flavonoids (the majority of them hydroxycinnamic acid esters of kaempferol and quercetin glycosides) and hydroxycinnamic acyl glycosides (with predominance of synapoyl gentiobiosides).

320nm

Fig 1. Peaks: (1) 3-caffeoylquinic acid; (2) 3-p-coumaroylquinic acid; (3) 3-feruloylquinic acid; (4) 4- caffeoylquinic acid; (5) kaempferol-3-O-(caffeoyl)-sophorotrioside-7-O-glucoside; (6) caffeoylferuloylquinic acid; (7) quercetin-3-O-(sinapoyl)-sophorotrioside-7-O-glucoside; (8) synapoyl glucoside; (9) kaempferol-3-O- (sinapoyl)-sophorotrioside-7-O-sophoroside; (10) kaempferol-3-O-(sinapoyl)-sophorotrioside-7-O-glucoside; (11) 4-p-coumaroylquinic acid; (12) kaempferol-3-O-(feruloyl)-sophorotrioside-7-O-glucoside; (13) disinapoyl- gentiobioside; (14) kaempferol-3-O-(sinapoyl)-sophorotrioside-7-O-rhamnoside; (15) quercetin-3-O-(sinapoyl)- sophorotrioside; (16) kaempferol-3-O-(methoxycaffeoyl)-sophorotrioside; (17) kaempferol-3-O-(feruloyl)- sophorotrioside-7-O-rhamnoside; (18) quercetin-3-O-sophoroside; (19) sinapoyl,caffeoyl,feruloyl-gentiobioside; (20) kaempferol-3-O-(caffeoyl)-sophorotrioside; (21) kaempferol-3-O-(sinapoyl)-sophorotrioside; (22) kaempferol-3-O-(feruloyl)-sophorotrioside; (23) kaempferol-3-O-sophoroside; (24) kaempferol-3-O- (methoxycaffeoyl/sinapoyl)-sophorotrioside-7-O-sophoroside; (25) sinapoyl,methoxycaffeoyl-gentiobioside; (26) kaempferol-3-O-(methoxicaffeoyl/sinapoyl)-sophorotrioside-7-O-glucoside; (27) kaempferol-3-O- (caffeoyl/sinapoyl)-sophorotrioside-7-O-sophoroside; (28) kaempferol-3-O-(caffeoyl/sinapoyl)-sophorotrioside-7- O-glucoside; (29) quercetin-3-O-(disinapoyl)-sophorotrioside-7-O-glucoside; (30) disinapoyl,methoxycaffeoyl- gentiobioside; (31) disinapoyl,caffeoyl-gentiobioside; (32) kaempferol-3-O-(disinapoyl)-sophorotrioside-7-O- glucoside; (33) disinapoyl-gentiobioside isomer; (34) sinapoyl,feruloyl-gentiobioside; (35) trisinapoyl- gentiobioside; (36) disinapoyl,feruloyl-gentiobioside; (37) sinapoyl,diferuloyl-gentiobioside.

111

Thermodynamic and Theoretical Study of Diphenylpyridines

Marisa A. A. Rocha a, Carlos F. R. A. C. Lima a, Lígia R. Gomes b and Luís M. N. B. F. Santos a

a Centro de Investigação em Química, Departamento de Química, Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre, 687, 4169-007 Porto, Portugal b CIAGEB-Faculdade de Ciências da Saúde, Escola Superior de Saúde da UFP, Universidade Fernando Pessoa, Rua Carlos Maia 296, P-4200-150 Porto, Portugal

The energetic and structural study of phenylpyridines has been a theme of research in our laboratory [1,2] due to their interesting electronic correlation capabilities. This work focuses the thermodynamic study of some diphenylpyridine isomers (Figure 1), in order to understand the relationship between structure and energetics.

N N N

IIIIII Figure 1. Schematic structural formulas of: (I) 2,6-diphenylpyridine; (II) 2,5-diphenylpyridine; (III) 3,5-diphenylpyridine.

Therefore, the compounds shown in figure 1, were synthesized using the Suzuki-Miyaura methodology.[3] The standard molar enthalpies of formation for the three isomers on crystalline state will be derived, at T=298.15 K by static bomb combustion calorimetry. The vapour pressures of the three isomers will be determined by a static apparatus based on a MKS capacitance diaphragm manometer recently described in the literature.[4] Trough the standard molar enthalpies of formation in the condensed phase and the standard molar enthalpies of sublimation, the standard molar enthalpies of formation in the gaseous phase will be derived for the three isomers. Computational chemistry (Density Functional Theory DFT) will be used in order to perform the geometry optimizations, energetic analysis and to derive the vibrational frequencies for the selected isomers.

References: [1] Ribeiro da Silva, M.A.V., Matos, M.A.R., Rio, C.A., Morais, V.M.F., Wang, J., Nichols, G. and Chickos, J.S. (2000), A Thermochemical and Theoretical Study of the Phenylpyridine Isomers, Journal Physical Chemistry, 104, 1774–1778. [2] Rocha, M. A., Low, J. N., Gomes, L. R., Quesada, A. and Santos, L. M. N. B. F. (2007), 2,6- Diphebylpyridine, Acta Crystallographica, E63, o4833. [3] Miyaura, N. and Suzuki, A. (1995), Palladium-Catalyzed Cross-Coupling Reactions of Organoboron Compounds, Chemical Reviews, 95, 2457-2483. [4] Monte, M. J. S., Santos, L. M. N. B. F., Fulem, M., Fonseca, J. M. S. and Sousa, C. A. D. (2006), New Static Apparatus and Vapor Pressure of Reference Materials: Naphthalene, Benzoic Acid, Benzophenone, and Ferrocene, Journal Chemical and Engineering Data, 51, 757-766.

Acknowledgements: Thanks are due for the financial support to Project POCI/QUI/61873/2004 supported by Fundação para a Ciência e Tecnologia (FCT) and by Programa Operacional Ciência e Inovação 2010 (POCI 2010), supported by the European Community Fund FEDER. C.F.R.A.C. Lima thanks also FCT and the European Social Fund (ESF) under the third Community Support Framework (CSF) for the award of a Ph.D. Research Grant (SRFH/BD/29394/2006). Presentation: POSTER

112

Form and Structure on Eduardo Souto de Moura’s architecture

B. Moreira1, C. Machado2

1 Faculty of Architecture, University of Porto, Portugal. 2 Faculty of Architecture, University of Porto, Portugal.

Regarding the subject “Form and Structure” we propose a study of Souto de Moura’s work through eight topics of architecture organized in a contradictory manner – by pairs – in order to create the necessary tensions and bring back the answers. Four chapters were born. The essay was not only the result of a theoretical investigation/discussion but also of a direct experimentation of some of the architect’s spaces. His practice and theory was then confronted with his own references (written and designed or built). Chapter 1 – Fragment and Unity – allowed us the possibility of showing how The Stijl’s influence was particularly evident in Souto de Moura’s early work: fragments of planes, textures and materials subsequently unified by means of an open space; but Aldo Rossi’s The Architecture of the City was also explored as well as Souto de Moura's vision, learned with Rossi, of the city as a sum of fragments and as an impossible unity. In Chapter 2 – Mass and Skeleton – we focused two distinct building systems explaining Souto de Moura’s affection for reinforced concrete structures; solid concrete walls and slabs relate to The Stijl’s architecture, but also to Northern Portugal’s building tradition of stone walls. The theme of the ruin, as memory and fragment, is also treated in this context. These mass structures are often complemented by a single metallic or concrete column, bringing together Souto de Moura and Álvaro Siza’s work; this chapter also deals with the problem of the building’s skin (and its misuse) related to Le Corbusier’s “Dom-Ino”. Chapter 3 – Evidence and Dissimulation – pretends to show how these topics are the architect’s everyday work – from the global plan to the detail. For Souto de Moura the details must have a minimum expression; complex building systems are therefore dissimulated, resulting in simple (not simplistic) forms, relating his work to Judd’s concept of “minimum”. The building itself may also have a minimum expression on its surroundings, being dissimulated as well, but the opposite can also be true: a building as a landmark. Evidence and Dissimulation also brings us the concept of truth – and a relation (by opposition) between Mies and Souto de Moura becomes evident. Finally, Chapter 4 – Artifact and Nature – confronts the building with its site. For Souto de Moura, the creation of architecture (a manmade Artifact) is the (re)creation of Nature in a way that one can’t be without the other. The theme of the ruin is very important as an evidence of a possible hybrid state between Artifact and Nature. In his first work, the ruin in the Gerês was neither created nor altered – it looks like it was stopped in time. Souto de Moura uses the ruin not only as a building material (reusing remains of old constructions for new purposes, as a fragment and as a memory) but also as a means of justifying the new Artifact (designing ruins when they are not present, as a false or invented fragment of memory). All these tensions lead us to a main conclusion: contradiction in Souto de Moura’s architecture appears not as a goal but as a result of the increasingly complexity of the architectural processes.

113

Possible correlation between morbid obesity and inflammation: a peripheral leukocyte study

A. Meneses1*, I. Maio1*, D. Pestana1, P. Freitas2, R. Monteiro1, I. Azevedo1, C. Calhau1, R. Soares1

1 Department of Biochemistry, Faculty of Medicine, University of Porto, Portugal. 2 Department of Endocrinology, S. João Hospital *The two authors contributed equally to the present study

Obesity has emerged as a major public health problem in the western world. Recently, more than an inactive bystander, adipose tissue is actually a dynamic and metabolically active one, since it releases a huge number of inflammatory cytokines [1,2]. Large size adipocytes mainly in visceral adipose tissue are prone to rupture, therefore evoking an inflammatory reaction [3]. Nuclear factor kappa-B (NF-kappaB), a transcription factor, is a primary event in inflammation. NF-kappaB is a crucial molecule regulating the expression and function of a wide spectrum of inflammatory genes [2]. Alkaline phosphatase (ALP) is a membrane- bound enzyme playing a role in lipids transport and storage, two processes occuring in adipose tissue [4]. In addition, ALP was reported to be associated with inflammation [5]. The purpose of our work was to examine the presence and activity of NFkappaB and ALP isoforms in peripheral leukocytes in morbid obese patients.

Blood samples were collected from 50 obese patients and 20 volunteers age- and sex- matched. Proteins were isolated from peripheral leukocytes by the TriPure method. ELISA assays were performed for NF-kappaB activity and intestinal and tissue-nonspecific isoforms of alkaline phosphatase (I-ALP and TNS-ALP) expression. Clinical data, such as weight, height, diet and BMI, were obtained and crossed with lab findings. Two subgroups of patients were identified: the first presenting NFkappaB activity mean values identical to controls and another exhibiting significantly higher NFkappaB activity. I-ALP and TNS-ALP isoforms expression were also increased in the subgroup presenting higher NFkappaB activity. In conclusion, two distinct groups of obese patients were identified according to the presence of inflammatory markers. Clinical parameters of these patients are now being evaluated in order to investigate the causes behind these distinct inflammatory profiles. References: [1] Charo IF, Ransohoff RM (2006) The many roles of chemokines and chemokine receptors in inflammation. N Engl J Med 354:610-621. [2] Stienstra R, Duval C, M 252 Ller M, Kersten S (2006) PPARs, Obesity, and Inflammation. PPAR Res 23;2007:95974. [3] Rosario Monteiro, Paulo M. S. T. de Castro, Conceição Calhau, Isabel Azevedo (2006) Adipocyte size and liability to cell death. Obes Surg 16(6):804-806. [4] Ali AT, Penny CB, Paiker JE, Psaras G, Ikram F, Crowther NJ (2006) The relationship between alkaline phosphatase activity and intracellular lipid accumulation in murine 3T3-L1 cells and human preadipocytes. Anal Biochem 15;354(2):247-54. [5] Shanmugham LN, Petrarca C, Castellani ML, Symeonidou I, Frydas S, Vecchiet J, Falasca K, Tete S, Conti P, Salini V (2007) IL-1beta induces alkaline phosphatase in human phagocytes. Arch Med Res 38(1):39-44.

114

In vitro and in vivo studies of the expression of carbohydrates in a canine mammary carcinoma cell line

J. Gomes 1, C. Lopes 2, E. Hellmén 3, C. Reis 1,4 and F. Gärtner 1,2

1 Institute of Molecular Pathology and Immunology of the University of Porto, Portugal; 2 Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal; 3 Department of Anatomy, Physiology and Biochemistry, Uppsala, Sweden; 4 Medical Faculty of Porto, Porto, Portugal.

Spontaneous mammary tumours are the most common neoplasia in the female dog and have a high biological and histomorphological heterogeneity. Approximately one-half of all mammary tumours in dogs are malignant and have a great potential to metastasize to the regional lymph nodes or other organs such as the lungs [1]. Malignant transformation is associated with abnormal glycosylation, resulting in the synthesis and expression of altered carbohydrates determinants. Although the majority of cancer research in humans is conducted using established cell lines, the interaction between the tumour and the host organism must be taken in consideration, so the results need to be confirmed using animal models. In order to study the biology of canine mammary tumours we used a previously established canine mammary cell line [2] and compared the information with an in vivo model. The CMT-U27 cell line, derived from a ductular carcinoma, was cultured and kept at 37ºC in 5% CO2 atmosphere. Cells were stained for expression of carbohydrates by immunoflurescence. In vivo experiments were performed using mice 6 weeks old of N:NIH(s)II-nu/nu strain. Tumours and organs which had been removed from these mice were fixed in 10% neutral-buffered formalin and embedded in paraffin for histopathology and immunohistochemistry studies. The CMT-U27 cells adhered to the bottom of the flask in single or paired cells as a compact thin monolayer. Immunoflurescence for carbohydrates showed reaction for anti- SLex, anti-Lex and anti-Lea antisera. The CMT-U27 cells grew when inoculated subcutaneously in the fat mammary pad of female nude mice. Tumour masses were histologically identical to the primary mammary tumour lesions, and when heterotransplanted tumours were re-cultured, the expression of carbohydrates was not altered. To look for metastatic targets tissues we performed an intravenous injection in the tail vein of the mice. These cells metastasised to lymph nodes, lungs, heart, spleen, kidney and liver. The pattern of expression of carbohydrates in the canine mammary carcinoma cell line suggests that these antigens could be useful as a prognostic tumour marker in canine mammary tumours. The aberrant expression of carbohydrates may also play a fundamental role in the molecular mechanisms of metastization to distant organs and facilitate positive interactions within the target organ. References: [1] Moulton, J.E. (1990), Tumors of the mammary gland. In Tumors in Domestic Animals, 3rd edn. Ed J.E. Moulton. Berkeley, University of California Press, 518-552.

[2] Hellmén, E. (1992), Characterization of four in vitro established canine mammary carcinoma and one atypical benign mixed tumor cell lines. In Vitro Cell Dev Biol, 28A, 309-19.

115

Determination of Cadmium Content in Sardine pilchardus

Ana C. Silva1,2, S. Morais1,*, A. Novo1, C. Luz1, E. Pinho2, S. Conceição2, C. Delerue- Matos 1 and M.B.P.P. Oliveira2

1 REQUIMTE/Instituto Superior de Engenharia do Porto, Rua Dr. António Bernardino de Almeida 431, 4200-072 Porto, Portugal. 2 REQUIMTE/Serviço de Bromatologia, Faculdade de Farmácia da Universidade do Porto, Rua Aníbal Cunha, 164, 4099-030 Porto, Portugal. *e-mail: [email protected]

Fish is nutritious and highly essential in a balanced diet, being an important source of proteins and lipids of high biological value, with long chain polyunsaturated fatty acids, and also liposoluble vitamins [1]. Fish can, on the other hand, be a source of contamination, because of the amounts of heavy elements they can contain. Heavy metals naturally occur in seawater in very low concentrations, but their levels have increased due to anthropogenic pollutants over time. Studies have shown that fish retain heavy metals from their environment [2]. Once heavy metals are accumulated by an aquatic organism they can be transferred through the upper classes of the food chain. Carnivores, including humans, obtain most of their heavy metal burden from the aquatic ecosystem by way of their food. Muscles are not always the best indicator of whole fish body contamination however, to judge about heavy metals fish safety, it is indispensable to analyze them since they are the edible parts. Cadmium has no beneficial effects in humans and there is no known homeostasis mechanism for it. Although toxicity and the resulting threat to human health of any contaminant are, of course, a function of concentration, it is well-known that chronic exposure to Cd at relatively low levels can cause adverse effects [2]. One of the most important source of Cd is dietary intake of seafood specially shellfish [2]. EU maximum level permitted for cadmium in fish fillets is 0.10 mg/kg wet weight [3]. The goal of this study was to determine the concentration of cadmium in the muscle of commercial Sardine (Sardine pilchardus) consumed in Portugal. To can attain this goal, methodologies based on microwave-assisted digestion coupled with graphite furnace atomic absorption spectrometry were developed. The results of this investigation suggest that the cadmium content in the edible tissues of sardines is below the maximum level established by the European Commission Regulation [3].

Acknowledgments The authors thank the Universidade do Porto for the financial support through the programme Investigação Científica na pré-graduação-Proj. Pluridisciplinares – Concurso 2007.

References: [1] Carvalho, M.L, Santiago, S. and Nunes, M.L. (2005), Assessment of the essential element and heavy metal content of edible fish muscle, Anal Bioanal Chem, 382, 426-432. [2] Dural, M., Göksu, M.Z.L. and Özak, A.A. (2006), Investigation of heavy metal levels in economically important fish species from the Tuzla Lagoon, Turkey, Food Chemistry, 102, 415- 421. [3] EU (2006), Regulation (EC) No. 1881/2006, JO L364, 20.12.2006, 5-24.

116

Optimization Study for Arsenic Quantification in Fish Using Microwave-assisted Digestion and Graphite Furnace Atomic Absorption Spectrometry

Ana C. Silva1,2, S. Morais1,*, A. Novo1, C. Luz1, E. Pinho2, S. Conceição2, C. Delerue- Matos 1 and M.B.P.P. Oliveira2

1 REQUIMTE/Instituto Superior de Engenharia do Porto, Rua Dr. António Bernardino de Almeida 431, 4200-072 Porto, Portugal. 2 REQUIMTE/Serviço de Bromatologia, Faculdade de Farmácia da Universidade do Porto, Rua Aníbal Cunha, 164, 4099-030 Porto, Portugal. *e-mail: [email protected]

Heavy metals from anthropogenic sources are increasingly being released into seawater, tend to accumulate in the biota and are subsequently transferred to humans through the food chain. It has been shown that water metals levels are positively correlated with concentrations in fish tissues [1]. Portugal is the biggest consumer of fish among all the European Union countries and one of the biggest in the world. Arsenic is toxic to human health even in trace amounts and the provisional tolerable weekly intake (PTWI) is 15 μg/kg of body weight per week [2]. The aim of this work is to develop an accurate and precise methodology based on closed vessel microwave-assisted digestion coupled with graphite furnace atomic absorption spectrometry (GFAAS) with Zeeman effect background correction for total arsenic determination in dry edible parts of fishes. In order to obtain the complete decomposition of the sample, the influence of sample weight, the oxidizing agents, temperature, pressure and time of digestion were optimised. Concerning the quantification of arsenic by GFAAS, the interferences were effectively eliminated by using Pd(NO3)2 as matrix modifier and by optimising the graphite furnace operating pyrolysis and atomization temperatures. Also, several quantifications methods were tested and compared.

Acknowledgments The authors thank the Universidade do Porto for the financial support through the programme Investigação Científica na pré-graduação-Proj. Pluridisciplinares – Concurso 2007.

References: [1] Has-Schön, E., Bogut. I and Strelec, I. (2006), Heavy metal profile in five species included in human diet, domiciled in the end of flow river Neretva (Croatia), Environmental Contamination Toxicology, 50, 545-551. [2] Alinnor, I.J. (2005), Assessment of elemental contaminants in water and fish samples from Aba river, Environmental Monitoring and Assessment, 102, 15-25.

117

Optimization of an enzymatic and chromatographic method for inorganic pyrophosphate based on firefly luciferase

Simone M. Marques1 and Joaquim C.G. Esteves da Silva1

1 Department of Chemistry, CIQ-UP, Faculty of Sciences, University of Porto, Portugal.

Inorganic pyrophosphate (PPi) is a compound generated mainly in anabolic pathways. Although PPi was initially regarded as a by-product its biological importance is now well established. In this communication a new method for quantifying PPi is described. This method is based on the production of ATP (and L - dehydroluciferin) from PPi and dehydroluciferyl-adenylate (L-AMP) catalyzed by firefly luciferase (EC 1.13.12.7) (Eq.1) [1].

L-AMP + PPi ⇌ L + ATP (1) The ATP formed can be identified and quantified by ion pair-HPLC with detection at 260 nm. The method was subjected to an optimization using experimental design methodologies to obtain suitable values for the identified factors: incubation time (tinc), inactivation time of the enzyme (tinac), pH of the assay (pH) and the concentrations of L-AMP (LAMP) and luciferase (Luc). Fig. 1 shows a response surface for the two observed critical factors: LAMP and tinc.

Fig.1 Response surface for the critical factors.

The optimized method is linear over a range of 0.1-10 μM PPi, with a limit of detection (LOD) of 1.2 μM and a limit of quantification (LOQ) of 3.9 μM.

Acknowledgements: Financial support from Universidade do Porto and Caixa Geral de Depósitos (Project IPG136) is acknowledged.

References: [1] Fraga, H., Fernandes, D., Novotny, J., Fontes, R. and Esteves da Silva, J.C.G. (2006), Firefly Luciferase Produces Hydrogen Peroxide as a Coproduct in Dehydroluciferyl Adenylate Fornation, ChemBioChem, 7 (6), 929-935.

118

Survey of the effects of the antibiotic minocycline on freshwater cyanobacteria

J.Pereira 1, M. S. Baptista 1,2, T. Stoichev 2, B. Heggie 3, V.M. Vasconcelos 2,4 and M.T.S.D. Vasconcelos 1,2

1 Department of Chemistry, Faculty of Sciences, University of Porto, Portugal. 2 CIMAR/CIIMAR, Centre of Marine and Environmental Research, University of Porto, Portugal. 3 HOVIONE FarmaCiencia, Portugal. 4 Department of Zoology and Anthropology, Faculty of Sciences, University of Porto, Portugal.

The monitoring of drug residues in the aquatic environment has gained much interest in recent years, as many pharmaceutical compounds can frequently be found in sewage treatment plant effluents and freshwater reservoirs at concentrations up to 5 μg L-1. Antibiotics are generally believed to be excreted unchanged by the body [1]. They are specifically designed to penetrate biological membranes and reach universal molecular systems, thereby increasing the probability of unexpected consequences for a number of species. Recently, it has been suggested that there is a need to develop ecotoxicity screening procedures that take into consideration the modes of action of therapeutic drugs on non-target species [2]. Minocycline hydrochloride, also known as minocycline, is a member of the tetracycline antibiotics class and has a very broad spectrum of action. The aim of this study is to determine the effects of minocycline on the growth of the cyanobacterium Microcystis aeruginosa Kütz., which commonly occurs in Portuguese freshwater systems. The strain was obtained from Torrão reservoir (Tâmega river) where it produces toxic blooms every year. To test the influence of the antibiotic, three concentrations (1, 10, 1000 μg L-1) were added to M. aeruginosa growing in f2 medium. The biological response will be assessed by the growth yield and vitality. The data will be used to estimate the lowest concentration capable of causing biological effects. The amount of prescribed minocycline, per year, in the European Union, will be used to determine if this concentration is likely to occur in Portuguese rivers. Preliminary results will be presented at the meeting. This work will contribute with new information on the effects of antibiotics on phytoplankton. The results obtained may be complemented with surveys of pharmaceutical compounds in Portuguese freshwater systems, enabling Portugal to face the challenges of an expected up-coming legislation. References: [1] Hircsh, R., Ternes, T., Haberer, K., Kratz, K.L. (1999), Occurrence of antibiotics in the aquatic environment, The Science of the Total Environment, 225, 109–118. [2] Daughton, C.G., Ternes, T.A. (1999), Pharmaceuticals and personal care products in the environment: agents of subtle change?, Environmental Health Perspectives, 107, 907–942.

Acknowledgements: This work was partially funded by Fundação para a Ciência e a Tecnologia, Portugal, (fellowships awarded to T. Stoichev (SFRH/BPD/32700/2006) and M. Baptista (SFRH/BD/16292/2004) and the CONC-REEQ/304/2001 re-equipment project) and Reitoria da Universidade do Porto, Portugal, through the “Investigação na Pré-Graduação” program.

119

Low-cost materials as adsorbents for Textile Wastewater Treatment

S. Santos, R. Deucher and R. Boaventura

Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal.

Dyeing processes generate high volumes of colored and toxic effluents. This kind of effluents is very difficult to treat. Conventional biological and physico-chemical treatments are not effective enough to remove color from wastewaters. Adsorption has been considered a simple and economical method. The use of activated carbon, the most widely used adsorbent, is limited by the high prices associated with its replacement and regeneration. The goal of this work is to investigate the utilization of natural and waste materials as alternatives to activated carbon on the decolorization of textile effluents. Several low-cost materials were firstly selected for the study: a Portuguese bentonitic clay from Alentejo, a commercial sepiolite clay, a calcinated waste sludge from a water treatment plant and an hydroxide metal waste sludge generated in a metal plating industry. Preliminary adsorption tests were conducted with these potential adsorbents and different kinds of dyes. The percentage of color removal was determined by absorbance measurement after one hour of contact time between colored solutions (30 mg/L of dye) and adsorbents (1.0 g/50 mL). Clay materials showed to be very effective for basic dyes removal (a total color removal was perceived). Sepiolite also showed a good performance for direct and sulfurous dyes adsorption. Waste sludge from the water treatment plant exhibited poor adsorption properties, but a good capacity for some basic and direct dyes was obtained. The metal hydroxide sludge, tested only with one dye of each class, showed to be a very effective material for this kind of application, namely for reactive, direct and acid dyes (100% color removal). Adsorption isotherms for some of the most promising dye/adsorbent systems were determined at neutral pH and 25 ºC and results were fitted to Langmuir and Freundlich models. Values in the range of 89.9 mg/g (Blue Reactive dye/hydroxide metal waste sludge) and 232 mg/g (Red Basic dye/bentonitic clay) were obtained as monolayer’s adsorption capacities given by Langmuir fittings. Further studies were conducted for Blue Reactive dye adsorption onto the hydroxide metal waste sludge. The influence of salts and dyeing auxiliary products (which appeared commonly in textile effluents) on the performance of adsorption was investigated. The results indicate that NaCl does not interfere significantly in the adsorption. Dyeing auxiliary chemicals and mainly Na2CO3 were responsible for a considerable decrease in the adsorbed amounts. The use of this kind of clays and the hydroxide metal waste sludge was then found to be very promising, respectively for basic and anionic textile dyes removal. Adsorption capacities obtained in this work are acceptable, in comparison with values reported in literature for commercial activated carbons and other alternative materials.

120

Determination of physical parameters in pharmaceutical powders by near infrared spectroscopy (NIR)

Ana Cruz 1,2, Sandra Soares1,2, Mafalda C. Sarraguça1, Paulo C. Costa2, Helena R. Amaral2, João A. Lopes1

1Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Portugal 2Serviço de Tecnologia Farmacêutica, Faculdade de Farmácia, Universidade do Porto, Portugal

Near infrared spectroscopy (NIR) can be used to determine physical and chemical parameters in solids, powders and liquids [1]. The application of this spectroscopic technique in the pharmaceutical industry is increasing [2]. NIR has been successfully applied for variability reduction in primary and secondary pharmaceutical production while as a research and quality control technique [3]. In the field of physical parameters determination, NIR spectroscopy is still underused. Parameters such as particle size, apparent volume and flowability, are defined by the Portuguese Pharmacopeia as some of the tests performed to characterize pharmaceutical powders. These tests require specific equipment and are time consuming. NIR spectroscopy can be used to determine these parameters, with the advantage of being a fast and low-cost technique. To establish the potential of the spectroscopic technique in the determination of some important physical parameters, a series of tests were performed in powder samples. The parameters determined were the angle of rest, apparent volume, and flowability. The tests were executed as determined in the pharmacopeia. The samples used were blended powders based on a pharmaceutical formulation of acetaminophen (as the active ingredient) and three excipients (microcrystalline cellulose, talc and magnesium stearate). The NIR spectrum was recorded on a Fourier Transform NIR analyzer in reflectance mode. Partial least squares (PLS) calibrations were optimized in terms of latent variables with cross-validation [4]. The NIR results were compared with the reference methods results and the performance of the NIR was accessed.

References [1] Burns, D.A. and Ciurczak, E.W. (2001) Handbook of Near-Infrared Analysis, Marcel Dekker, 2nd Edition, New York. [2] Blanco, M. Coello, J., Iturriaga, H., Maspoch, S. and Pezuela, C.(1998) Near-Infrared Spectroscopy in the Pharmaceutical Industry, Analyst, 123, 135R. [3] Lopes, J.A., Costa, P.F., Alves, T.P. and Menezes, J.C. (2004) Chemometrics in Bioprocess Engineering: Process Analytical Technology (PAT) Applications, Chemom. Intell. Lab. Syst., 74, 269. [4] Martens, H. and Næs, T. (1989) Multivariate Calibration, John Wiley & Sons, Chicester.

121

An expeditious method for determination of residues of organochlorine pesticides in sediments and its application to Portuguese estuarine areas

P. N. R. Rodrigues1,2,*, P. N. Carvalho1,2, F. Alves1,2, R. Evangelista1,2, M. C. P.Basto1,2 and M. T. S. D.Vasconcelos1,2

1 Department of Chemistry, Faculty of Sciences, University of Porto, Portugal. 2 CIMAR/CIIMAR – Centre of Marine and Environmental Research, University of Porto, Portugal. * [email protected]

Organochlorine pesticides (OCPs) integrate the semi volatile persistent organic compounds (POPs) that exhibit potentially harmful effects to the environment [1]. As most of the POPs, OCPs are lipophilic, persist in various media and some can be transported over long distances to regions where they have never been used [2]. As OCPs can be introduced into the aquatic environment and accumulated in sediments by several pathways, levels of OCPs in sediments should be determined and controlled whenever possible. Analysis of OCPs implicates extraction of these compounds from the sediment, which is difficult owing to strong interaction among OCPs and different constituents of the sediments, particularly organic content. A method involving a combination of microwave assisted extraction, headspace solid- phase microextraction (HS-SPME) and gas chromatography coupled to mass spectrometry (for identification and quantification) or gas chromatography with electron capture detection (for quantification of previously identified OCPs) with complete automation of the HS-SPME procedure, which increases the sample throughput, was optimized. To minimize matrix effects, a step for elimination of sulphur interference was included when required and standard addition was used for calibration purpose. Method validation was performed by means of analysis of ten OCPs in reference sediments with certified values: (a) MetranalTM 16, from Analytika Ltd. (Prague, Czech Republic); and (b) CNS300-04-100 from Resource Technology Corporation (Salisbury, United Kingdom). The method resulted effective, relatively simple and fast, and it was applied for monitoring residues of OCPs from estuarine sediments of different Portuguese rivers. In this communication, the characteristics of the method will be presented as well as the nature and levels of OCPs found in estuarine areas of Portuguese rivers. References: [1] Zhou, R., L. Zhu, and Q. Kong (2007), Persistent chlorinated pesticides in fish species from Qiantang River in East China, Chemosphere, 68, pp. 838-847. [2] Li, Y.F. and Macdonald, R.W. (2005), Sources and pathways of selected organochlorine pesticides to the Arctic and the effect of pathway divergence on HCH trends in biota: a review Science of The Total Environment, 342, pp. 87-106. Acknowledges: This work was partially funded by Portuguese Foundation for Science and Technology (fellowship awarded to Rafael Evangelista (SFRH/BD/20163/2004), the CONC-REEQ/304/2001 re- equipment project and POCTI/CTA/48386/2002 project).

122

Kinetics of Inhibition of Firefly Luciferase by Oxyluciferin and Dehydroluciferyl-adenylate

César Ribeiro1 and Joaquim C.G. Esteves da Silva1

1 Department of Chemistry, CIQ-UP, Faculty of Sciences, University of Porto, Portugal.

Firefly luciferase (Luc) catalyzes the oxidation of D-luciferin (LH2) with molecular oxygen in the presence of ATP and Mg2+ with emission of yellow-green light in an extremely efficient process [1]. This reaction involves the formation of an enzyme bound adenylyl intermediate (LH2-AMP) (Eq. 1) following oxidation with release of AMP, pyrophosphate, CO2 and oxyluciferin (Eq. 2), the assumed light emitter and also an inhibitor of luciferase. Dehydroluciferyl-adenylate (L-AMP), a potent inhibitor of light emission, is also formed as a side product of the bioluminescence reaction. Luc + LH2 + ATP → Luc·LH2-AMP + pyrophosphate (1)

Luc·LH2-AMP + O2 → Luc + AMP + CO2 + oxyluciferin + photon (2) The light emission starts with an initial flash that quickly decays to a low basal level due to accumulation of inhibitory products. So far no conclusive explanation has been proposed regarding the kinetic mechanism of inhibition and the relative importance of the inhibitory products. The applications of luciferase include a remarkable number of biochemical and clinical tools, many of which use the luciferase gene as a reporter of gene expression and regulation. Luciferase is also a very sensitive analytical tool for detection of ATP. The current study provides a detailed kinetic model of the inhibition exerted by oxyluciferin and L-AMP by measuring the light production in the presence and absence of these inhibitors (Fig. 1). Nonlinear regression analysis suggest that oxyluciferin is a competitive inhibitor of luciferase (Ki = 1 – 3 μM) while L-AMP act as a tight-binding competitive inhibitor (Ki = 2 nM).

1500000 200000 0 uM Oxyluciferin 0 nM L-AMP 2.5 uM Oxyluciferin 40 nM L-AMP 5.0 uM Oxyluciferin 150000 1000000 60 nM L-AMP

100000 RLU RLU 500000 50000

0 0 0.0 0.5 1.0 1.5 2.0 2.5 0.0 1.0 2.0 3.0 4.0 t/s t/s Fig.1. Inhibitory effect of oxyluciferin and L-AMP on firefly luciferase bioluminescence. Mixtures cointaining ATP 50 μM and LH2 10 μM were injected into solutions of Luc. 10 nM in hepes buffer (pH=7.5) pre-incubated with inhibitor. References: [1] McElroy, W.D., Seliger, H.H. and White, E.H. (1969), Mechanism of Bioluminescence, Chemiluminescence and Enzyme Function in the Oxidation of Firefly Luciferin, Photochemistry and Photobiology, 10 (3), 153-170.

123

Effect of pyrophosphate on firefly luciferase bioluminescence

Filipe Peralta1, Joaquim C.G. Esteves da Silva1 and Rui Fontes2

1 Department of Chemistry, CIQ-UP, Faculty of Sciences, University of Porto, Portugal. 2 Serviço de Bioquímica (U38-FCT), Faculty of Medicine, University of Porto, Portugal.

Firefly luciferase (LUC, EC 1.13.12.7) is an enzyme that catalyses the oxidation of firefly luciferin (LH2) giving rise to light [1]. The bioluminescence reaction involves the formation, from LH2 and ATP, of an enzyme-bound adenylyl intermediate (LUC•LH2- AMP) and its subsequent oxidation with release of AMP, CO2 and oxyluciferin in an electronically excited state.

LUC + LH2 + ATP ⇌ LUC•LH2-AMP + PPi (1) LUC•LH2-AMP + O2 → LUC + AMP + CO2 + oxyluciferin + photon (2)

Inorganic pyrophosphate (PPi) was described as stabilizer and/or activator of firefly bioluminescence [2]. However, PPi effect is both intriguing and paradoxical: since PPi is a product of the adenylation step (reaction 1), an inhibitory effect would be expected and can indeed be observed but, as stated, PPi can also act as a stabilizer or activator of LUC bioluminescence.

In this communication we show that the stabilizing and activating effect of PPi on LUC bioluminescence can be explained by the theory proposed by McElroy et al [2]. This theory proposes the pyrophosphorolysis of the enzyme-formed inhibitor dehydroluciferyl- adenylate (L-AMP) giving rise to dehydroluciferin (L) and ATP (Eq. 3) - L is a much less powerful inhibitor than L-AMP.

LUC•L-AMP + PPi ⇌ LUC + L + ATP (3)

Consequently, this reaction converts L-AMP into L allowing the removal of a powerful inhibitor (L-AMP) from the enzyme active site and its replacement by a less powerful inhibitor (L).

Acknowledgements: Financial support from Universidade do Porto and Caixa Geral de Depósitos (Project IPG136) is acknowledged.

References: [1] Nakatsu, T., Ichiyama, S., Hiratake, J., Saldanha, A., Kobashi, N., Sakata, K., and Kato, H. (2006) Structural basis for the spectral difference in luciferase bioluminescence. Nature 440, 372-376

[2] Rhodes, W. C., and McElroy, W. D. (1958) The synthesis and function of luciferyl-adenylate and oxyluciferyl-adenylate. J Biol Chem 233, 1528-1537.

124

Molecular Energetics of Hydroxybenzaldehyde Isomers

Mariana Vidinha, Manuel J. S. Monte, Maria D. M. C. Ribeiro da Silva

Centro de Investigação em Química, Departamento de Química, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre, 687, P-4169-007 Porto, Portugal.

The thermochemical study of different classes of aromatic compounds has been carried out in our Research Group (CIQ-UP) in order to correlate molecular energetic data with the structural characteristics of the molecules. Some of our attention has been particularly devoted to Schiff bases derived from ketones or aldehydes with diamines, which being molecules whose role is well known on the development of agrochemical and pharmaceutical industries, particularly in the context of catalysis, have motivated our interest.

In this context, during the development of the energetic study of some Schiff bases, derived from salicylaldehyde (2-hydroxybenzaldehyde) with aliphatic and cyclic diamines, we detected the lack of the relevant thermochemical data for that phenol derivative. The need of the knowledge of such parameters led us to study that compound [1]. Considering that thermochemical data for phenols with carbon-bonded substituents containing the acyl group are scarce, as it is reported by Slayden and Liebman [2], we decided to extend the study to the other two isomers of the salicylaldehyde (3-hydroxybenzaldehyde and 4- hydroxybenzaldehyde, Fig. 1), which we present in this work. O

O C H

C H

OH OH

(a) (b)

Figure 1: Structural formula for (a) 3-hydroxybenzaldehyde and (b) 4-hydroxybenzaldehyde.

The standard (pº = 0.1 MPa) molar enthalpies of formation, in gaseous phase, at T = 298.15 K, for 3- and 4- hydroxybenzaldehyde were derived from measurements of standard molar energies of combustion, using a static bomb calorimeter, and from the standard molar enthalpies of sublimation, determined by Knudsen method effusion experiments. These results allow the derivation of the values of the enthalpies of formation of the two hydroxybenzaldehyde isomers and, consequently, to establish structural correlations with these energetic data.

[1] - Ribeiro da Silva, M. D. M. C., Araújo, N. R.M. (2007), J. Chem. Thermodynamics 39, 1372 – 1376.

[2] – Slayden, S. W., Liebman, J. F. (2003), Therchemistry of Phenols and Related Arenols, in: Z. Rappoport (Ed.), The Chemistry of Phenols, Wiley, Chichester, (Chapter 3).

125

Touristic Topographies : transversality and authenticity in an Portuguese context

João Nuno Pinto Marques

Architecture Degree Final Diploma | Faculty of Architecture, University of Porto, Portugal,

The essay Touristic Topographies is a reflection about the ways that tourism, while activity that crosses several multidisciplinary fields (where the architecture plays an important role), develops itself in territorial terms and about the produced effects, either on environmental levels (built areas) or social levels, that emerges along with its growth.

A first analysis seeks to contextualize the first transitory movements and the social changes that were at the origin of tourism, along with the respective dynamics and transversalities that took place in those territories. With basis on the different reasons that took man to travel by free will and with no economic needs, we are able to observe the evolution of the tourist profiles through time – pilgrim, traveler, sightseer, vacationer, post-tourist – and its importance in the definition of the actual touristic industry guidelines.

In a second part, we tried to identify distinct touristic models existents in the Portuguese context and analyze it according with patterns that include densities, uses, effects and transversalities produced on the different territories. In this way, we identified five different major patterns that illustrate the main strategies used by the touristic managements in terms of urban development: concentration I (coastal lines) concentration II (traditional cities), nodulation, stimulation and dissemination. However, the used method does not enclosure or predicts all existent models, as it can not be taken as isolated objects, attending that this models can derivate one from the others, emerge simultaneously, coexist in a same place or be developed in different scales. Instead, it is proposed some action lines and information forms organized and orientated towards the Portuguese example.

A third part of the essay concerns the aesthetics and the authenticity of the architecture in the touristic models. The potential of the architecture is analyzed as a touristic atractor and a reflection is made on the rhetoric in which it is based on, attending that the tourism is, above all, an economic activity that seeks the profit as the main goal. In these terms, the role and the theoretical basis of the architecture behind the touristic models is questioned, attending that the tourism is being developed according to the whims of a globalized economy, that has contributed to the dilution of identity values and to the profusion of an aesthetic obtained from instruments as the simulation or the iconization. It is now common to find a mix of cultures, styles and constructive techniques associated to some touristic models that contributes to an uncharacterized and anachronic type of architecture in order to obtain a certain image or thematic.

An alternative option can come out from an interpretation of the real identity, cultural and historic values according to the reality where they are inserted and through a conscientious integration of it in the current time. During this transformation process, the architecture can obtain a very delicate and specific role insuring the authenticity.

126

Sensor for amperometric determination of ammonia in seawater

C. Peixe1,2, C.M. Pereira1, F. Silva1 and M. T. Borges2,3

1Department of Chemistry, Faculty of Science, University of Porto, Portugal. R. Campo Alegre, 687, 4169 – 007 Porto, Portugal 2 Ciimar-University of Porto; Rua dos Bragas 289, 4050-123 Porto, Portugal. 3 Faculty of Sciences (Zoology-Anthropology Dep.)-University of Porto; Praça Gomes Teixeira, 4099-002 Porto, Portugal.

The present work reviews the development of amperometric ammonia determination by the use of an interface between two immiscible electrolyte solutions (ITIES). Cyclic voltammetry and square wave voltammetry have been applied to the analysis of the voltammograms for transfer of ammonium cation facilitated by dibenzo-18-crown-6 [1]. The liquid-liquid interface was supported by use of silicon membrane containing micropores. The organic solvent used in this work was 2-nitrophenyloctylether (NPOE). The organic phase was prepared dissolving bis(triphenylphosphoranylidene)-ammonium tetrakis(4-chlorophenyl)borate (BTPPATPBCl) and 1,3:2,4_dibenzylidene sorbitol (DBS) (gelling agent) in the organic solvent. NPOE/DBS gels were prepared accordingly to the literature [2]. The aqueous phase was prepared dissolving lithium chloride and adding different amounts of ammonium chloride. Figure 1 shows cyclic voltammograms obtained for microITIES using different aqueous phases.

LiCl 4,0x10-7 LiCl+NH4Cl

2,0x10-7 I /A 0,0

-2,0x10-7

-300 -200 -100 0 100 200 300 400 500 600 φ /mV

Figure 1 – Cyclic voltammograms obtained in different conditions.

+ In the presence of the ionophore the addition of NH4 to the aqueous solution results in the presence of a peak at -200 mV. The peak current intensity increase can be related with the concentration of ammonium cation in the solution.

Acknowledgements: CIIMAR for the project EC-CRAFT/016869/2006, “Raceways – A Hypertensive fish farming concept for lasting competitiveness and superior production” and financial support from FCT.

References: [1] Liao, Y., Okuwaki, M., Kitamura, F., Ohsaka, T. and Tokuda, K., Electrochimica Acta, 44 (1998), 117-124. [2] Silva, F., Sousa, M.J., Pereira, C., Electrochimica Acta, 42 (1997), 3095-3103.

127

Copper(II)/Ciprofloxacin complexes: Synthesis and Solution Studies

C. Queirós , I. Sousa, E. Pereira, P.Gameiro

1 Requimte, Department of Chemistry, Faculty of Sciences, University of Porto, Portugal.

Fluoroquinolones are a group of antibacterial agents used currently against a wide variety of infections. Currently, these drugs are known to have two prime enzyme targets in the bacterial cell, DNA gyrase and topoisomerase IV and that they inhibit these enzymes by stabilizing the DNA–DNA gyrase complex and/or the DNA–topoisomerase IV complex. Although, to some extent, all fluoroquinolones are active against these enzymes, exposure to the drugs induces mutations in both proteins promoting higher levels of bacterial resistance and modifications to the fluoroquinolones based on structure-activity- relationships (SARs) have been made to achieve lower level of resistance. Synthesis of fluoroquinolone metal complexes has been carried out as some antibacterial activity studies show that these complexes allowed the alteration of the potency and specificity of these antibiotics. In this work the synthesis of complex copper(II)/ciprofloxacin has been achieved and the stability constants of this system, in aqueous solution determined, by UV/visible spectrophotometry. Briefly, spectrophotometric pH titrations were performed in stock solutions of different metal/ligand molar ratio – 1:1 and 1:2 (1.25x10- 5–2.5x10-5M) – for the binary species, and aliquots of strong acid or base were added to adjust pH to the desired value. Spectra were recorded at 25.0 ºC in 1 cm quartz cuvettes with a slit width of 1.5 nm in the range 225 – 450 nm. The equilibrium constants defined by Eqs. (1) and (2): pM + qH2L + rH ⇔ MpLqHr (1) p q r βpqrs = [MpLqHr] / [M] [H2L] [H] (2)

(where M is metal, H2L is ciprofloxacin, in the cationic form and H is proton were refined by least-squares calculation using the computer program pHab taking into account the presence of the hydroxide species of copper and the autoprotolysis of water.

The pKa values of the ciprofloxacin are similar to those previously reported for several oyher fluoroquinolones and as was expected from their chelating properties they form very stable complexes with copper(II). The values of the stability constants determined for binary copper(II) complexes are very high and similar to those found in literature for other fluoroquinolones. Due to the stability of the complexes at physiological pH they can be suitable metalloantibiotic candidates. Acknowledgements: Partial financial support for this work was provided by FCT through POCI/SAU-FCF/56003/2004. PG thanks Bayer for ciprofloxacin.

128

Thermochemical studies of two cyclic ureas: Imidazolinone and N,N'-Trimethyleneurea

V. Freitas1, M. Roux2, J. Elguero3, M. Ribeiro da Silva1

1 CIQ-UP, Department of Chemistry, Faculty of Science, University of Porto, Portugal. 2 Instituto de Química Física "Rocasolano", C.S.I.C., Serrano, 119, E-28006 Madrid, Spain. 3 Instituto de Química Médica, CSIC, Juan de la Cierva, 3, E-28006 Madrid, Spain.

Due to the significance of urea derivatives both as drugs and as materials, its importance does not need to be stressed too much [1,2]. Some of us have studied abiotic receptors designed for interacting with ureas [3,4]. To discuss the reactivity of the compounds where those molecular units are present, we feel useful to know their thermodynamic properties, althougt we discovered that almost none was known about cyclic ureas. Thus efforts of our Research Groups were joint to determine some gaseous enthalpies of formation, following previous studies for other class of relevant nitrogen heterocyclic molecules. A survey of the literature on thermochemical properties of cyclic ureas yield only one hit and it concerns the recently published study of benzimidazolinone [5].

H N NH

O O

N NH H a) b) Figure 1. Structural formula for a) N,N´-ethyleneurea (2-imidazolidone) and b) N,N´-propyleneurea (N,N´- trimethyleneurea)

We will report now the experimental results of the energy of combustion and enthalpy of sublimation obtained for imidazolinone and N,N'-trimethyleneurea (represented in Fig. 1), using static bomb calorimetry and Knudsen effusion technique, respectively. These data allow to derive the values of the standard molar enthalpies of formation, in the gaseous state, for both compounds. The results are discussed in terms of structural contributions.

[1] http://en.wikipedia.org/wiki/Urea

[2] Bagley, M. C. et al., Org. Biomol. Chem. 2006, 4, 4158.

[3] Claramunt, R. M. et al., Biosens. Bioelectron. 2004, 20, 1242.

[4] Herranz F., et al., Tetrahedron Lett. 2006, 47, 9017; and references cited therein.

[5] Morais, V. M. F., et al., Mol. Phys. 2006, 104, 325.

129

Feeling Architecture. The importance of ‘the other’ senses in the experience of space.

S. Silva Natária

Faculty of Architecture, University of Porto, Portugal.

This abstract refers to my final dissertation2 at F.AU.P. Based on a phenomenological approach to architecture, it focuses on the perception of space by Touch, Smell and Hearing, asking questions that enable the creation of a multisensorial architectonic language: Which are the non-visual sensory stimuli that participate in the user’s experience of architecture? What composition principles do they materialize? What role do they play in that experience? To reinforce my own sensory awareness and to better understand the possibility of provoking aesthetic pleasure independently from the sense of Sight, I made an incursion in blind user’s experience of space by conducting a series of interviews. An inquest to the users of Koolhaas’s Casa da Música and Siza Vieira’s Museu de Serralves showed how two different architectonic languages were also perceived differently by ‘the other’ senses, causing different reactions: the first, a desire for physical sensorial interaction, the second a more visual attitude of contemplation. This multisensorial approach to architecture’s creative process led me to the creation of a sensorial data-base, a recollection and organization of tactile, sound and smell stimuli: textures, consistencies and temperatures of walls and pavements of different materials, sounds of steps and fountains, and aromatic vegetation. From the analysis of all data gathered, I concluded that, to create a multisensorial language, architects must control the sensory parameters of shape, temperature, texture, consistency, smell and sound of every element chosen for a certain space. These non-visual stimuli can be assembled together according to composition principles also used for visual stimuli, like rhythms, gradations, harmonies and contrasts. In addition, they also accumulate functions of architectonic identity, helping to identify and distinct spaces, and of spatial orientation, attracting, directing, delimitating and punctuating the user’s way trough the building. There is scope for further work. It is critical to test these conclusions with a wider survey, covering staff and blind users of the buildings cited above. This investigation intends to open the practice of architecture to a creative challenge of adapting project processes and formal languages to the creation of spaces that explore the potential of architecture’s intrinsic multisensoriality. Its results give us, architects, a more complete notion of how users perceive the built environment, so that we can make well informed choices and better decisions during the creation process. And finally, a basic knowledge that should be part of any architect’s education: how to address our buildings aesthetic messages to all human senses, creating buildings that, like persons, objects or movies, are memorable for their own unique perfume, touch or soundtrack.

[1] Natária, Sara Silva, Sentir a Arquitectura. A importância dos ‘outros’ Sentidos na leitura da obra arquitectónica, Prova Final de Licenciatura em Arquitectura, F.A.U.P. 2005.

130

Deep Eutectic solvents: A new kind of Ionic Liquids

R. Costa, F. Silva and C.M. Pereira

Department of Chemistry, Faculty of Science, University of Porto, Portugal. R. Campo Alegre, 687, 4169 – 007 Porto, Portugal

Ionic liquids (ILs) are salts with melting point below 100°C and have recently attracted considerable attention as potential alternatives to conventional organic solvents in a variety of synthetic, catalytic, and electrochemical applications [1]. Deep eutectic solvents (DES) are a new type of these ionic liquids and are obtained by mixing a large range of quaternary ammonium salts with metal salts, amides, carboxylic acids or alcohols. Attractive features of these liquids such the potential to reduce pollution in industrial processes have led to investigations as alternative reaction media for a variety of application that usually use organic solvents. Eutectic solvents can be use in catalysis, electropolishing, metal extraction and metal deposition (e.g. Cr) [2,3]. Chromium plating is a massive industry and all of them use chromic acid technology. Conventional chromic acid plating processes are extremely hazardous because they use hexavalent chromium, which is highly toxic and carcinogenic. Choline chloride:chromium chloride (III).6H2O (cromeline) based ionic liquids allow the use of trivalent chromium less toxic [4].

1.0x10-4

5.0x10-5

0.0

-5

I/A -5.0x10

-4 -1.0x10 scan rate: 5 mV/s 20 mV/s -1.5x10-4 50 mV/s 100 mV/s

-2.0x10-4 -1.0 -0.8 -0.6 -0.4 -0.2 0.0 E vs SCE/V Figure 1 – Cyclic voltammogram measured at Hg/cromeline interface.

A cyclic voltammetry experiment, as in other controlled potential experiments, a potential is applied to the system, and the faradaic current response is measured (a faradaic current is the current due to a redox reaction) [5]. This technique was applied to study the Hg/cromeline interface. The work was completed with the study of nucleation that occurs on mercury surface by chronoamperometry. Data will be presented and discussed later. References: [1] A. P. Abbott, G. Capper, S. Gray; Chem. Phys. Chem., 2006, 7, 803 – 806. [2] A. P.Abbott, G. Capper, K. Mckenzie, K. Ryder; Electrochimica Acta, 2006, 4420 – 4425. [3] A .P. Abbott, K. J. Mckenzie; Phys. Chem. Chem. Phys., 2006, 8, 4265 – 4279. [4] A. P. Abbott, G. Capper, D. Davies, R. Rasheed; Chem. Eur. J, 2004, 10, 3769 – 3774.

[5] A. J. Bard, L. R. Faulkner; Electrochemical methods – Fundaments and applications, 2nd edition, Wiley&Sons, Inc., N.Y., 2001.

131

Authenticity evaluation and molecular characterization of Portuguese varieties of Juglans regia using microsatellite markers

E.M. Cabral 1, M.A. Faria1 and M.B.P.P. Oliveira1

1 REQUIMTE, Serviço de Bromatologia, Faculty of Pharmacy, University of Porto, Portugal.

Persian walnut is cultivated mostly in Asia, Europe and America. Europe accounts for 23% of walnut overall production. In 2005, Portugal was responsible for an area of cultivation of 3155 ha producing 4565 t of nuts. The main cultivated varieties are of French and American origin, however, the Portuguese cultivars Arco and Rego have a significant expression. Worldwide cultivars have been characterized at the molecular level however there is a complete lack of this kind of studies for Portuguese varieties. Several methodologies have been used to evaluate the genetic diversity and relationships among cultivars of walnut, including isozymes, restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD), inter-simple sequence repeats (ISSR), and simple sequence repeats (SSR). These studies reflect the need for accurate cultivar identification and for the verification of paternities and genealogy. SSRs are the markers of choice for genotyping since they are highly polymorphic, co-dominant and relatively easy to use. In this study 21 cultivars of walnut collected in an experimental field of Direcção Regional de Agricultura e Pescas do Centro (DRAPC) (including three Portuguese varieties Arco, Rego and Samil) were analyzed at 14 microsatellite loci [1,2]. DNA was obtained through the CTAB method [3]. The genetic relations were evaluated and a database of microsatellite profiles was constructed to be used in nut and nut based products authenticity. In order to obtain DNA extracts directly from the nuts, two other methods of DNA extraction (Wizard® Plus Minipreps DNA Purification System, Promega and GenElute Plant Genomic DNA Miniprep Kit, SIGMA) were tested and compared with the CTAB method.

Microsatellite markers used showed useful in distinguishing all cultivars under study, with the exceptions of Franquette and Ronde de Montignac. These results are in agreement with the bibliography that considers these markers the most appropriate to this kind of studies. The genotypes of the Portuguese varieties were compared with the ones of international varieties using the neighbor-joining algorithm and with those results it’s possible to associate the variety Rego with the group of varieties of French origin and Samil with the North-American ones.

References: [1] Foroni, I. et al. (2007), Identification of ‘Sorrento’ walnut using simple sequence repeats (SSRs), Genet Resour Crop Evol, 54, 1081-1094. [2] Dangl, G.S. et al. (2005), Characterization of 14 microsatellite markers for genetic analysis and cultivar identification of walnut, J Amer Soc Hort Sci, 130, 348-354. [3] Wang, X-D., Wang, Z-P., Zou, Y-P. (1996), An improved procedure for the isolation of nuclear DNA from leaves of wild grapevine dried with silica gel, Plant Mol. Biol. Rep., 14, 369-373.

132

UV PROTECTION OF GEOTEXTILES BY CHIMASSORB 944

1 1 2 2 Inês Valente , Paulo Almeida , José Ricardo Carneiro and Maria de Lurdes Lopes

1 Department of Chemistry, Faculty of Science, University of Porto, Portugal. 2 Department of Civil Engineering, Faculty of Engineering, University of Porto, Portugal.

Geosynthetics are polymeric materials widely applied in construction of infrastructures (landfills, roads, tunnels, railways, dams) due to economical, technical and environmental advantages. Geotextiles are the most applied geosynthetics, as they can perform functions like separation, filtration, drainage, protection and reinforcement. In those applications, geotextiles can be exposed to several agents (such as, UV radiation and other weathering agents, atmospheric oxygen, high temperatures, acids or alkalis and microorganisms) that may affect their durability and, consequently, their useful lifetime. The degradation suffered by these materials is often retarded and/or inhibited by the incorporation of chemical additives, such as UV stabilisers and antioxidants in their composition. So, it is extremely important to study these compounds during the degradation process of the geotextiles. In this work, it was developed a new methodology for the determination of the additive Chimassorb 944 (C944) in polypropylene (PP) geotextiles. C944 is an additive widely used in the stabilisation of polyolefins against the damaging effects of UV radiation and oxidation induced by heat. The methodology developed for the determination of C944 can be divided in two main steps: (1) ultrasonic extraction with chloroform at 60 ºC for removing C944 from the geotextile sample; (2) separation and determination of the additive by liquid chromatography with UV detection at 244 nm. This method was used to monitor the level of C944 in geotextile samples exposed to UV radiation in a laboratory weatherometer (the QUV). Unstabilised PP geotextiles were also exposed to UV radiation in the QUV. Morphologic changes on the PP fibres of the geotextiles were characterised by scanning electron microscopy (SEM). C944 proved to be important in the stabilisation of the PP geotextiles against UV radiation (the unstabilised PP geotextiles were destroyed after UV-ageing tests in the QUV). However, the level of C944 in stabilized geotextiles decreased during UV radiation exposure. SEM analysis showed that polymeric structure of the geotextiles exposed to higher UV radiation was more damaged.

133

Computational Analisys of Cynara cardunculus L. Cardosins Pomoters

Alberto M. Pessoa1, Jorge Teixeira1 and José Pissarra1

1 Department of Botany, School of Sciences, University of Porto, Portugal

Cardosins are aspartic proteinases from Cynara cardunculus L., a plant used to act as a milk-clotter for the manufacture of cheese. To further understand the role of these proteinases in the plant and how the transcriptional regulation occurs, the promoter regions from 4 Cardosin genes were analysed and compared in silico. The promoter region of the Cardosins A-like (A, C and D) and B, previously isolated and sequenced, were analysed using PlantCARE (a database of plant cis-acting regulatory elements, enhancers and repressors). Perl scripts were used to parse the reports obtained from PlantCARE for each Cardosin sequence and to sort and format the results, so that they could be displayed on pDraw32 (Acaclone Software), for easy interpretation. The data was also converted into a Microsoft® Excel™ spreadsheet, using a similar procedure, in order to assemble charts illustrative of the TATA Box motif distribution along the sequences. The plausibility of the results was then verified by comparing the set of motifs obtained for each of the cardosins with previous studies. The analysis with PlantCARE revealed that Cardosin B has the least variety of cis-acting elements, having one exclusive light responsive element and sharing 4 motifs with the Cardosins A-like (high transcription levels conferring element, MeJA- responsiveness involved element and a MYB binding site involved in drought- inducibility). Cardosins A-like yielded a greater variety of results, having in common 3 motifs: abscisic acid responsiveness and light responsiveness involved elements, and the MYB binding site also present in Cardosin B. However, the analysis revealed distinct features on their promoters: Cardosin A – the presence of a heat stress responsiveness involved element and the lack of light responsive elements; Cardosin C – the presence of an ethylene-responsive element, and 3 different light responsive elements, while no anaerobic induction, MeJA-responsiveness and endosperm expression involved elements were detected; Cardosin D – the presence of an exclusive light responsive element and the lack of high transcription levels conferring element and circadian control involved element. The TATA Box motif distribution along the sequences was also analysed and showed that the TATA box is likely to be in a distinct position, being further upstream from the ATG codon in Cardosin B promoter, when compared to the Cardosins A-like promoters. These results suggest that, apart from being similar, the promoters of the 4 Cardosins may have different metabolic sensitivities. Therefore, their differential expression and detection could be possible. These observations are being studied by promoter fusions to reporter genes.

134

Hydrogen Peroxide Reversible Luminescence Sensors

Célia Rocha and Joaquim C.G. Esteves da Silva

Department of Chemistry, CIQ-UP, Faculty of Sciences, University of Porto, Portugal.

The determination of reactive oxygen species (ROS) is a matter of great importance in the study of numerous physiological and pathological processes. The production of ROS, which includes hydrogen peroxide, is mediated by multiple oxidative stress mechanisms responsible for changes in the cells and tissues. Beside this role in these pathological processes they are also involved in numerous physiological process of regulation in the organism. With the objective to develop luminescent sensors for in vitro and in vivo measurement of hydrogen peroxide two reversible detection methods are being optimized [1,2]. A direct assay of hydrogen peroxide at neutral pH using the Eu(III)-tetracycline complex. This complex in aqueous solution binds to hydrogen peroxide to form a fluorescent complex with increased fluorescence at 613 nm when is excited at 400 nm.

The second method is an H2O2 sensor based on the room-temperature phosphorescence of the nanocomposite TiO2/SiO2. The TiO2/SiO2 composite was prepared by the sol-gel route and produces phosphorescence at about 600 nm when excited at 400nm (Fig. 1) – in the presence of hydrogen peroxide the phosphorescence emission is quenched.

Int. 596 10000

5000

0 400 600 800 Wavelength (nm)

Fig. 1 – Phosphorescence emission spectra (Ex. 400 nm) of the TiO2/SiO2 composite.

This communication will present and discussed the results of these two reversible detection methods for hydrogen peroxide.

Acknowledgements: Financial support from Fundação para a Ciência e Tecnologia (Lisboa) (FSE-FEDER) (Project PTDC/QUI/71001/2006) is acknowledged.

References: [1] Durkop, A., Wolfbeis, O.S., (2005), Nonenzimatic Direct Assay of Hydrogen Peroxide at Neutral pH using the Eu3Tc Fluorescent Probe, Journal of Fluorescence, 15 (5), 755-761.

[2] Shu, X., Chen, Y., Yuan, H., Gao, S. and Xiao, D. (2007), H2O2 Sensor Based on the Room- Temperature Phosphorescence of Nano TiO2/SiO2 Composite, Anal. Chem, 79 (10), 3695-3702.

135

Chromium remediation using zero valent iron - batch studies

G. Carvalho 1,2, Aurora Silva1, Cristina Delerue-Matos1 , A. Vega y Fuente2,3 and António Fiúza 3

1 REQUIMTE, Instituto Superior de Engenharia do Instituto Politécnico do Porto 2Departamento de Geotecnia do Instituto Superior de Engenharia do Instituto Politécnico do Porto 3 CIGAR, Faculdade de Engenharia da Universidade do Porto

One of the main problems that modern societies are facing today, as a direct consequence of their activities, is the change of the original conditions of the sites and their components as result of human activities. One of the group of substances that require special attention is heavy metals. They are considered a priority problem due to its toxicity, and in certain cases, its carcinogenic potential when in contact with populations or ecosystems. Chromium is one of the top pollutants, because of their relative abundance as a contaminant, the dangerousness of their ions, in particular Cr6+ and its high mobility in the environment. In fact, as it occurs in anionic form it is not significantely sorbed by the soil. This work aims to contribute for the knowledge of the geo-environmental phenomenon associated to remediation of Cr6+ in contaminated water by using zero valent iron. The net reactions of Cr6+ reduction whit zero valent iron and co precipitation of Cr3+ and Fe2+ are [1]:

In this work a low coast commercial iron was used to establish reaction kinetic parameters, using batch test.

Solutions of Cr6+ were prepared with potassium dichromate. The method used to determine the concentration of chromium present in samples was the colorimetric method [2].

Batch kinetic experiments were performed to evaluate the removal rate of Cr6+ in the presence of iron under different operating parameters: temperature, concentration, pH, iron mass. These tests allowed to establish that reaction follows a first order kinetic. The kinetic constants and the activation energy were estimated from the experimental data. AKNOWLEDGMENS The authors would like to thank Fundação para a Ciência e Tecnologia for the financial support of the project POCI/ECM/59779/2004 References [1] Biswajit Mukhopadhyay, Jon Sundquist, Rodney J. Schmitz, Removal of Cr(VI) Cr- contaminated groundwater through electrochemical addition of Fe(II) Journal of environmental Management 82 (2002) 66-76. [2] APHA Standard Methods for the Examination of Water and Wastewater 20th edition (1998), Method 3500-Cr (D) - Colorimetric Method.

136

Development of a multicommutated flow system for chemiluminometric quantification of gentamicin

L. Santos1, A. Araújo1, A. Fachini1, M. Montenegro1, B.F. Reis2

1 REQUIMTE, Department of Physical Chemistry, Faculty of Pharmacy, University of Porto, R. Aníbal Cunha, 4050 Porto, Portugal. 2 Centro de Energia Nuclear na Agricultura, Universidade de S. Paulo, PO Box 96, 13400-970, Piracicaba- SP

Multicommutation is a continuous flow system technique that presents several advantages relatively to flow injection analysis (FIA), such as versatility, low reagent consumption, and robustness [1]. Chemiluminescence is a high sensitivity methodology, with low detection limits that requires simple instrumentation. It is based on intensity measurements of light produced in chemiluminescent reaction. The signal intensity can be correlated with the concentration of any of the starting reagents. This analytical technique allows the quantification of a wide variety of compounds that are directly or indirectly involved in the chemiluminometric reaction [2]. The aim of this work is to quantify gentamicin in different matrices using a multicommutated flow system where the chemiluminometric oxidation of luminol by hypochlorite is accomplished. In the presence of gentamicin, an aminoglycoside antibiotic that is effective against a wide variety of gram-positive and gram-negative bacteria, the hypochlorite concentration is reduced since it oxidizes the secondary amines presents in the chemical structure of the analyte. The optimization process had been separeted in two parts: optimization of chemical conditions and optimization of flow conditions. In the first part, parameters such as the hypochlorite and luminol concentrations and the pH of the buffer solution used to prepare these reagents have being studied. After that, the flow conditions will be tested. In the final, the proposed method will be applied to the quantification of gentamicin in pharmaceutical formulations and foodstuffs. References: [1] Lavorante, A.F., Feres, M.A. and Reis, B.F. (2006), Multi-commutation in Flow Analysis: A Versatile Tool for the Development of the Automatic Analytical Procedure Focused on the Reduction of Reagent Consumption, Spectroscopy Letters, 39 (6), 631-650. [2] García-Campaña, A.M. and Baeyens, W.R.G. (2000), Principles and recent analytical applications of chemiluminescence, Analusis, 28 (8), 686-698. Acknowledgements: L. Santos thanks to Fundação Astrazeneca the master degree scholarship conceded.

137

Potentiometric determination of gentamicin in pharmaceutical formulations

Martina Kocia, Célia Amorimb, Alberto Araújob ,Maria da Conceição Montenegrob and Marie Pospíšilova, Petr Solich

a Department of Analytical chemistry, Fakulty of Pharmacy, Charles University in Hradec Králové, Heyrovského 1203, Hradec Kralove 500 05, Czech Republic bREQUIMTE, Departamento de Química-Física, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha, 164, 4099-030 Porto, Portugal. email: [email protected]

Gentamicin is an aminoglycoside antibiotic, and can treat many types of bacterial infections, particularly Gram-negative infection. It is one of the heat-stable antibiotics that remain active after autoclaving, which make it particularly useful in the preparation in the certain microbiological growth media. Traditionally, pharmaceutical analysis of these compounds relies heavily on chromatography, yet also frequently requiring the use of reagent-based detection principles like spectrophotometric [1,2],chemiluminescent detection in flow-injection analysis [3], capillary electrophoresis [4] a novel LC/ELSD [5], thin-layer chromatography [6], HPLC [7].A simple and sensitive potentiometric procedure for the determination of gentamicin is presented, after development of selective electrodes based on PVC membranes using different type of cyclodextrins and its derivatives, or Evans Blue complex with gentamicin as ionophers. Several mediator solvents and lipophilic ionic species were also used in order to optimize the characteristics of the membranes. The evaluation of the general characteristics for all the units prepared revealed an analytical linear range of about 2x10-4; 5x10-3 mol L-1, slopes of about 50 mV/dec, good reproducibility and low response times. The evaluation of Potentiometric selectivity coefficients will permit to choose the best electrode for gentamicin control in pharmaceutical preparations. References [1] El-Didamony, A. M.m Amin, A.S., Ghoneim, A.K., Telebany, A.M., (2006), Indirect spectrophotometric determination of gentamicin and vancomycin antibiotics based on their oxidation by potassium permangate, Central European J. of Chem. 4 (4), pp 708-72 [2] Ryan, J.A, (1984), Colorimetric determination of gentamicin, kanamycin, tobramycin, and amikacin aminoglycosides with 2,4-dinitrofluorobenzene, J. of Pharm. Sci. 73 (9), pp. 1301-1302 [3] Ranmos-Fernandez, J.M., Garcia-Campana, A.M.,Ales-Barrero, F.,Bosque-Sendra, (J.M, 2006), Corrigendum to “Determination of gentamicin in pharmaceutical formulations using peroxyoxalate chemiluminescent detection in flow-injection analysis, Talanta 69 (3), pp. 763-768 [4] Garcia-Ruiz, C., Marina,M.L., (2006), Recent advances in the analysis of antibiotics by capillary electrophoresis, Electrophoresis 27 (1), pp. 266-282 [5] Megoulas, N.C., Koupparis, M.A., (2004), Development and validation of a novel LC/ELSD method for the quantitation of gentamicin sulphate components in pharmaceuticals, J. Pharm. Biom. Anal. 36 (1), pp. 73- 79 [6] Sekkat, M., Fabre, H., Simeon De Bouchberg,M.m Mandrou, B., (1989), Determination of aminoglycosides in pharmaceutical formulations – I. Thin-layer chromatography, J. Pharm. Biom. Anal 7 (12), pp.883-892 [7]Fabre, H., Seddat, M., Blanchin, M.D., Mandrou, B., (1989), Determination of aminoglycosides in pharmaceutical formulations – II. High-performance liquid chromatography, J. Pharm. Biom. Anal 7(7), pp. 1711-1718 Aknowledgement: I would like to thank program Erasmus and MSMT for financial support.

138

Influence of organic pollutants on phytoremediation of copper by a salt marsh plant

A. C. M. Dias1,2*, C. M. R. Almeida2, A. P. Mucha2 and M. T. S. D.Vasconcelos1,2

1 Department of Chemistry, Faculty of Sciences, University of Porto, Portugal. 2 CIMAR/CIIMAR - Centre of Marine and Environmental Research, University of Porto, Portugal * [email protected]

Metal-contaminated soil can be remediated or cleanup by chemical, physical or biological techniques [1]. Plants may participate in decontamination either directly, by the uptake of pollutants and their immobilization, degradation or transformation within the plant, or indirectly by creating a rhizosphere environment (region where the soil and the roots of the plants come in contact) which promotes degradation by bacterial or fungal species. Sites contaminated with trace metals are frequently and unfortunately also contaminated with organic pollutants of different families. The simultaneous presence of pollutants of different groups might cause antagonisms or synergisms effects on the biological remediation processes, because different chemicals may interact among themselves and with the organisms present at rhizosphere, thus changing remediation efficiency. Studies of metal accumulation in plants from estuarine sediments contaminated with a mixture of inorganic and organic pollutants are scarce, although deserve investigation because estuaries are often considered sinks for varied pollutants. This study aims to investigate the effect of some organic compounds which are frequently present in polluted environments, like PAHs, pesticides and organotins, on the uptake of metals, like copper (Cu), by the salt marsh plant Halimiones portulacoides. This plant was selected because it is commonly found in Portuguese temperate salt marshes and is capable of accumulating several metals, including Cu [2]. For this purpose, ex-situ and in hydroponics experiments are being carried out using sediment and water from a salt marsh from Cávado River estuary. Sets of plants collected at the same site are being exposed to known concentrations of both Cu and a specific organic pollutant, during 6 days. Afterwards, Cu will be determine in the media and in plant dried tissues (roots, stems and leaves) by atomic absorption spectrometry, after digestion assisted by high pressure microwave. Whether the results indicate antagonistic or synergistic effects, the simultaneous presence of pollutants of different families should be taken in consideration in future studies of suitability of plants for remediation, including phytostabilization. References: [1] McEldowney S., Hardman D. J., Waite S. (1993). Treatment technologies, in S. McEldowney S., Hardman D. J., Waite S (Eds.). Pollution, ecology and biotreatment, Singapore pp. 48-58, Longman Singapore Publishers Pvt. Ltd. [2] Caçador, I., Vale, C., Catarino, F., (2000). Seasonal variation of Zn, Pb, Cu and Cd concentrations in the root-sediment system of Spartina maritima and Halimione portulacoides from Tagus estuary salt marshes. Marine Environmental Research 49, 279-290.

Acknowledgement: This work was partially funded by Fundação para a Ciência e Tecnologia (FCT), Portugal, through project POCTI/CTA/48386/2002.

139

DFT STUDY ON THE ABILITY OF CALIX[2]FURANO[2]PYRROLE TO FORM HOST-GUEST COMPLEXES WITH DIFFERENT IONS

C.A. Teixeira dos Santos, and A.L. Magalhães

Department of Chemistry, Faculty of Science, University of Porto, Portugal.

The applications of calixarene derivatives as new receptors have attracted considerable interest in the area of host-guest chemistry[1]. The presence of sulfur or nitrogen atoms in the ring linkages or upper rim seems to greatly enhance the complexation abilities toward transition metal ions, when compared to the classical calixarene systems. Applications of these molecules as new receptors, building blocks and /or molecular platforms have attracted considerable interest in a wide range of areas such as host-guest chemistry, separation chemistry, environment protection, selective ion transport and sensors [1]. Recently, DFT methods have been applied successfully in the study of the structural features and conformational equilibrium of sulfur bridged derivative compounds[2]. This work presents a comparative study of the complexation ability of calix[2]furano[2]pyrrole toward different positive ions. The B3LYP/SDD[3] level was used to estimate the binding energy of the host-guest 1:1 complexes between the cone conformer of calix[2]furano[2]pyrrole (1) and the alkaline ions Li+, Na+, K+, Rb+, Cs+, the alkaline-earth ions Be2+, Mg2+, Ca2+, Sr2+, Ba2+, and also the transition metal ions Ag+, Cu2+, Zn2+, Hg2+ and Au3+ as well. The theoretical predictions are compared with available experimental data[3]. Structural and electronic characteristics are presented, in order to support some experimental results and to discuss the different ability of the ligand to form complexes with the ions under study.

NH O

O NH

(1) References: [1]Lhoták P., Eur. J. Org. Chem., 2004, 1675-1692. [2]Suwattanamala A., Magalhães A.L., Gomes, J.A.N.F., Theor. Chem. Acc., 2007,117, 431-440 [3]Lim S.M., Chung HJ, Paeng K, Lee C., Choi H.N., Lee W., Anal. Chim. Acta., 2002, 453, 81- 88.

140

Fiber optic pH sensors based on CdTe quantum dots

Helena Gonçalves1, Conceição Mendonça1, Joaquim C.G. Esteves da Silva1, César Maule2 and Pedro Jorge2

1 Department of Chemistry, CIQ-UP, Faculty of Sciences, University of Porto, Portugal. 2 Optoelectronics Unit of INESC Porto, Department of Physics, University of Porto, Portugal.

Quantum dots (QDs) are gaining a great scientific interest due to their tunable optical (absorption and emission of radiation) properties. Also, by modifying the surface of QDs with hydrophilic capping ligands provides stability in water. One end of these hydrophilic capping ligands contains a thio group that binds with the QDs surface, and the other, polar end contains a carboxylic group that makes them water soluble. Because the carboxylic acid group has acid-base properties the optical properties of QDs may be used as pH sensing molecular devices. In this communication the synthesis of thiglycolate capped CdTe QDs is described and their luminescence properties (excitation and emission wavelength and lifetimes) obtained as function of the reaction time, concentration and pH are presented. Moreover, the immobilization of QDs in fiber optics using sol-gel techniques is described and their use as fiber optic pH sensors assessed. Fig. 1 shows the emission spectra of the synthesized CdTe QDs as a function of the reaction time (from 15 minutes and up to 27 hours reaction time) (a) and as function of the pH (from 3 to 9). The effect of the reaction time is to shift the emission wavelength of the CdTe QDs to higher wavelengths (from yellow/ green up to red) and by changing the reaction time the emission wavelength can be tuned. Decreasing the pH provokes a shift towards higher wavelengths. Also, the lifetimes are reaction time and pH dependent. These results suggest that the CdTe QDs have great potential as fiber optics pH sensors.

60000 40000 Int. 50000 30000 40000 t.

In 30000 20000

20000 10000 10000 0 0 400 500 600 700 500 550 600 650 700 750 Wavel ength (nm) λ Emission/ nm a. b. Fig. 1 – Fluorescence emission spectra of QDs as function of the reaction time (a) and pH (b).

Acknowledgements: Financial support from Fundação para a Ciência e Tecnologia (Lisboa) (FSE-FEDER) (Project POCTI/QUI/44614/2007) is acknowledged. References: [1] Alivisatos, A.P. (1996), Semicondutor cluster, nanocrystals and quantum dots, Science, 217, 933-937.

141

Steady state and lifetime chemical sensors based on Ruthenium complexes

Helena Gonçalves1, Joaquim C.G. Esteves da Silva1, César Maule2 and Pedro Jorge2

1 Department of Chemistry, CIQ-UP, Faculty of Sciences, University of Porto, Portugal. 2 Optoelectronics Unit of INESC Porto, Department of Physics, University of Porto, Portugal.

In recent years molecular recognition and supramolecular host-guest interaction have attracted much attention [1]. The design of highly selective and sensitive receptors for metal ions is of great importance in the area of ion transport, chemosensing, biomedical and environmental applications, especially if the sensor is portable, allowing in situ measurements. Ru(II) complexes with α-diimine ligands are particularly suited for luminescence based sensing due to their high quantum yields, long lifetimes and compatibility with low cost optoelectronics. In this communication the results of two Ru(II) complexes will be presented. The first complex, (Ru(Phen)2Dcbpy)Cl2, is constituted by the ligands 1,10-phenantroline (Phen) and 2,2’-bipyridine-4,4’-dicarboxilic acid (Dcbpy) and the second complex, (Ru(Ph2phen)2Dcbpy)Cl2, has 4,7-difenil-1,10-Fenantrolina (Ph2phen) and 2,2’-bipyridine- 4,4’-dicarboxilic acid (Dcbpy), which are pH sensitive. It was found that the first complex has a pKa of 3 and an operation range of 2.5 pH units. Its lifetime at the pKa was τpka= 325 ns changing by ≈172 ns over the full operation range, which is well suited for frequency domain interrogation (Fig. 1). For the complex second complex it was determined a pKa of 2 and a range of 2.5 pH units was observed. Its τpka was 470 ns changing by ≈160 ns over the whole operation range. 670 8000 660 7000 Peak wavelength (nm) wavelength Peak

6000 650

5000 640

4000 630 Luminescence Intensity (a.u.) Intensity Luminescence 3000

620 123456789 pH

Fig. 1 - Intensity (•) and peak wavelength (°) pH titration curves obtained for (Ru(Phen)2Dcbpy)Cl2 complex.

Acknowledgements: Financial support from Fundação para a Ciência e Tecnologia (Lisboa) (FSE- FEDER) (Project POCTI/QUI/44614/2007) is acknowledged. References: [1] Clarke Y., Xu W., Demas J.N. and DeGraff B. (2000), Lifetime-Based pH sensor system based on a polymer-supported Ruthenium(II) complex, 72 (15), 3468-3475.

142

Molecular characterization of the Portuguese patients with Mucopolysaccharidosis IIIC: two novel mutations in the HGSNAT gene

Coutinho MF1,2, Lacerda L2, Prata MJ1,3, Ribeiro H2, Lopes L2, Ferreira C2 and Alves S2

1 Department of Zoology & Anthropology, Faculty of Sciences, University of Porto, Portugal. 2 Centro de Genética Médica Jacinto Magalhães, INSA I.P., Porto, Portugal. 3 IPATIMUP, Porto, Portugal

Mucopolysaccharidosis IIIC (Sanfilippo Syndrome C), is a lysosomal storage disorder caused by the inherited deficiency of the lysosomal membrane enzyme acetyl–coenzyme A: α-glucosaminide N-acetyltransferase (N-acetyltransferase), which leads to impaired degradation of heparan sulfate. The gene that encodes this enzyme -HGSNAT- has been recently identified [1,2]. Here we report on the molecular analysis of the HGSNAT gene in the 3 Portuguese patients diagnosed with MPS IIIC that lead to the identification of two novel mutations: an insertion c.525-526InsT and a splicing mutation IVS3-2A→G, which is responsible for the skipping of exon 4. Both mutations give origin to transcripts that lead to the synthesis of truncated non- functional proteins. However, since the STOP codons appear very prematurely, these transcripts are probably degraded by nonsense mediated mRNA decay. Furthermore, having detected that c.525-526InsT accounts for 83% of the mutant alleles in our patient series might be of great epidemiological relevance. Since there is no known consanguinity among the patients here studied, c.525-526InsT can represent a founder mutation in the context of the Portuguese population and consequently it must be considered as a primary target in molecular studies of MPS IIIC in Portugal. The strategies developed for mutation analysis, in this study, constitute valuable tools that will allow carrier detection and prenatal molecular diagnostics, leading to the improvement of genetic counselling with great benefits for the Sanfilippo C families.

References:

[1] Fan, Xiaolian, Zhang H, Zhang S, Bagshaw RD, Tropak MB, Callahan JW, Mahuran DJ. (2006) Identification of the Gene Encoding the Enzime Deficient in Mucopolysaccharidosis IIIC (Sanfilippo Disease Type C) The Journal of Human Genetics 2006 October; 79: 738-744.

[2] Hřebíček M., Mrázová L, Seyrantepe V, Durand S, Roslin NM, Nosková L, Hartmannová H, Ivánek R, Cízkova A, Poupetová H, Sikora J, Urinovská J, Stranecký V, Zeman J, Lepage P, Roquis D, Verner A, Ausseil J, Beesley CE, Maire I, Poorthuis BJ, van de Kamp J, van Diggelen OP, Wevers RA, Hudson TJ, Fujiwara TM, Majewski J, Morgan K, Kmoch S, Pshezhetsky AV (2006), Mutations in TMEM76 that cause Mucopolysaccharidosis IIIC (Sanfilippo C Syndrome) The American Journal of Human Genetics 2006 November; 79L, 807-819.

143

Scopoletin, a natural coumarin isolated from Agrostistachys gaudichaudii (Euphorbiaceae)

C. Silva 1, A.E. Oliveira 1, A.P. Almeida 1,2, H. Cidade 1,2, A. Kijjoa 3,4 and M.S.J. Nascimento 2,5

1 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal. 3 Department of Chemistry, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Portugal. 4 Centre of Marine and Environmental Research (CIIMAR), University of Porto, Portugal. 5 Department of Microbiology, Faculty of Pharmacy, University of Porto, Portugal.

Euphorbiaceae plants are a well known source of natural products with different scaffolds (terpenoids, flavonoids, alkaloids, coumarins, and tannins) showing several interesting biological activities namely antitumor [1,2], and anti-inflammatory [3]. One of the less studied genus of this family is Agrostistachys, which comprises eight or nine species distributed from several countries of Asia, including Thailand [2]. In the course of our research on bioactive secondary metabolites from medicinal plants we have isolated one coumarin from Agrostistachys gaudichaudii: scopoletin (Fig. 1). This compound was isolated by chromatographic techniques (CC and TLC) and the structure elucidation was achieved by spectroscopic methods especially high field NMR (HSQC and HMBC) and mass spectrometry. To the best of our knowledge this is the first report of scopoletin in this species. HO O O

CH O 3 Fig. 1 - Structure of scopoletin. References: [1] Puapairoj, P., Naengchomnong, W., Kijjoa, A., Pinto, M.M., Pedro, M., Nascimento, M.S.J., Silva, A.M.S. and Herz, W. (2005), Cytotoxic Activity of Lupane-Type Triterpenes from Glochidion sphaerogynum and Glochidion eriocarpum Two of Which Induce Apoptosis, Planta Medica, 71, 208-213. [2] Choi, Y.-H., Pezzuto, J.M., Kinghorn, A.D. and Farnsworth, N.R. (1988), Plant Anticancer Agents, XLVI. Cytotoxic Casbane-Type Constituents of Agrostistachys hookeri, Journal of Natural Products, 51 (1), 110-116. [3] Paya, M., Ferrandiz, M.L., Erradi, F., Terencio, M.C., Kijjoa, A., Pinto, M.M. and Alcaraz, M.J. (1996), Inhibition of Inflammatory Responses by a Series of Novel Dolabrane Derivatives, European Journal of Pharmacology, 312, 97-105. Acknowledgements: FCT (I&D, nº226/94), FEDER, POCI, U. Porto, and Caixa Geral de Depósitos for financial support.

144

5-(Hydroxymethyl)-2-furfural, a natural compound isolated from Strophioblachia fimbricalyx Boerl. (Euphorbiaceae)

K.C. Marques 1, A.E. Oliveira 1, A.P. Almeida 1,2, H. Cidade 1,2, A. Kijjoa 3,4 and M.S.J. Nascimento 2,5

1 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal. 3 Department of Chemistry, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Portugal. 4 Centre of Marine and Environmental Research (CIIMAR), University of Porto, Portugal. 5 Department of Microbiology, Faculty of Pharmacy, University of Porto, Portugal.

Strophioblachia fimbricalyx Boerl. (Euphorbiaceae) is a small shrub up to 1.5 m high native in northeast Thailand. Megastigmanes and flavone glycosides were isolated from the aerial parts of this species [1]. As a part of continuing search for novel bioactive substances, we are investigating the chemical composition of roots of S. fimbricalyx Boerl., that were collected on Prachuab Kirikan Province (Thailand). The extract purification led to the isolation of 5-(hydroxymethyl)-2-furfural (Fig. 1), a known substance with antitumoral activity [2]. The structure determination was based on spectroscopic data (1H NMR, 13C NMR, HSQC and HMBC) and was according with the literature [3]. To the best of our knowledge this is the first report of 5-(hydroxymethyl)-2-furfural in this species.

HO H

O O

Fig. 1 – Structure of 5-(hydroxymethyl)-2-furfural

References:

[1] Kaewkrud, W., Otsuka, H., Ruchirawat, S. and Kanchanapoom, T. (2008), Megastigmane and Flavone Glycosides from Strophioblachia fimbricalyx Boerl., Journal of Natural Medicines, 62 (1), 124-125.

[2] Michail, K., Matzi, V., Maier, A., Herwig, R., Greilberger, J., Juan, H., Kunert, O. and Wintersteiger, R. (2007), Hydroxymethylfurfural: An Enemy or a Friendly Xenobiotic? A Bioanalytical Approach, Analytical and Bioanalytical Chemistry, 387 (8), 2801-2814.

[3] Mizushina, Y., Yagita, E., Kuramochi, K., Kuriyama, I., Shimazaki, N., Koiwai, O., Uchiyama, Y., Yomezawa, Y., Sugawara, F., Kobayashi, S., Sakaguchi, K. and Yoshida, H. (2006), 5- (Hydroxymethyl)-2-furfural: A Selective Inhibitor of DNA Polymerase λ and Terminal Deoxynucleotidyltransferase, Archives of Biochemistry and Biophysics, 446 (1), 69-76.

Acknowledgements: FCT (I&D, nº226/94), FEDER, POCI, U. Porto, and Caixa Geral de Depósitos for financial support.

145

New approaches for estimating the postmortem interval based on vitreous humor trace element levels

H. Andrade1, A. Almeida1, S. Frazão2, T. Magalhães2 and A. Santos2

1 REQUIMTE, Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Portugal. 2 Serviço de Medicina Legal, Faculdade de Medicina, Universidade do Porto, Portugal. Serviço de Patologia Forense, Instituto Nacional de Medicina Legal, I.P. – Delegação do Norte.

The estimation of the postmortem interval (PMI), i.e., the time since death, is a main issue in the field of forensic science and legal medicine [1]. Most of the currently employed methods use body temperature-based algorithms, which are subject to considerable inaccuracy [2]. In the last 60 years, numerous methods for estimating PMI based on concentrations changes in body fluids have also been proposed. The postmortem increase of potassium in vitreous humor (VH) has been the most studied, and several formulae have been proposed for estimating PMI based on it [3]. However, the correlation between K concentration in VH and PMI depends on several factors (e.g., cause of death, previous electrolyte imbalance, etc.), which influences the accuracy of PMI estimation. Recently, studies with animal models (rabbits) have shown that changes of several trace element levels in VH could correlated with PMI within different intervals after dead and could represent a reliable alternative strategy for estimating PMI [4-5]. The work we have been done is intended to address this topic: to look for more accurate methods for the determination of PMI based on VH trace element levels. Using an analytical methodology based on Inductively Coupled Plasma-Mass Spectrometry (ICP- MS) technique, samples of VH collected from human bodies at different times since death (3-30 h) were analyzed. From the tested elements (Ag, Al, As, Sb, Cr, Ba, Be, Co, Cu, Cs, Cd, Mo, Mn, Ni, Li, Fe, Sr, Pb, Rb, Tl, V, Zn, Si, I and Ti), some showed promising for the intended purpose, specially Rb, which showed a consistent non-linear increase in all the cases studied (n=8). References: [1] Henssge, C. and Madea, B. (2007), Estimation of the time since death, Forensic Science International, 165(2-3), 182-184. [2] McDowall, K.L., Lenihan, D.V., Busuttil, A., Glasby, M.A. (1998), The use of absolute refractory period in the estimation of early postmortem interval, Forensic Science International, 91(3), 163-170. [3] Madea, B. (2005), Is there recent progress in the estimation of the postmortem interval by means of thanatochemistry?, Forensic Science International, 151(2-3), 139-149. [4] Xu, X.M., Gong, Z.Q. and Sun, Y.G. (2002), Study on the relationship between PMI and the concentration of magnesium and iron in the vitreous humor of rabbit after death, Fa Yi Xue Za Zhi, 18(2), 65-66. [5] Gong, Z.Q., Xu, X.M., Zeng, X.B., Sun, Y.G. and Wang, D.W. (2001), Study on the relationship between PMI and the concentration of zincum and nickel in the vitreous humor of rabbit after death, Fa Yi Xue Za Zhi, 17(3):129-131. [6] Gong, Z.Q., Zeng, X.B. and Sun, Y.G. (2002), Study on the relationship between PMI and the concentration of 21 elements in vitreous humor of rabbit after death, Fa Yi Xue Za Zhi, 18(2):67-69.

146

New approaches to the diagnostic of death by drowning based on trace element levels in blood of the cardiac cavities

F. Pereira1, T. Magalhães2, A. Almeida1, A. Santos2 and B. Santos3

1 REQUIMTE, Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Portugal. 2 Serviço de Medicina Legal, Faculdade de Medicina, Universidade do Porto, Portugal. Serviço de Patologia Forense, Instituto Nacional de Medicina Legal, I.P. – Delegação do Norte. 3 Serviço de Patologia Forense, Instituto Nacional de Medicina Legal, I.P. – Delegação do Centro.

Investigation of bodies recovered out of water is a common medico-legal task. However, the key question whether the victim died due to drowning or was simply put into the water after death can frequently not easily be solved [1]. The difficulties are related to the few macroscopic findings seen during the autopsy examination, usually non-specific or doubtful. Additionally, the putrefaction, frequently present, increases the uncertainty of the diagnostic. Several approaches to support the diagnosis of death by drowning have been investigated. Beyond the classical detection of diatoms in blood and tissues, one of those approaches may be the determination of trace elements (TE) in blood, using highly sensitive analytical techniques. This is based on the following principle: several TE typically found in water are not present (or are present at much lower concentration) in human body. Therefore, in the case of death by submersion in water, those TE will become increased in the body. In this context, one of the elements that has gained broad acceptance is strontium (Sr), since it is, from the studied TE, the one that shows the higher ratio [Sr] in water / [Sr] in body fluids or tissues. For example, for seawater, the ratio [Sr] in water / [Sr] in blood is ca. 267, which makes it a highly useful indicator [2]. Additionally, some studies have also shown that a significant difference can be found between the Sr concentration in the left ventricle ([Sr]LV) and the right ventricle ([Sr]RV) blood, and the use of the two variables ([Sr]LV and [Sr]LV-[Sr]RV) have been proposed. However, the usefulness of Sr determination in case of deaths by submersion in freshwaters is not so clear, because of the lower Sr concentrations in these waters [3]. In the last few years, a highly sensitive instrumental technique, the so-called Inductively Coupled Plasma-Mass Spectrometry (ICP-MS), has gained increased importance in forensic studies. Our current work regards its utilization in the determination of TE in blood of the cardiac cavities and the water where the corpses were found in order to increase the certainty of the diagnostic of death by drowning. Main specific objectives are the evaluation of the difference between Sr and other TE concentrations in LV and RV blood as a marker of drowning and the identification of other TE (beyond Sr) that could be an advantageous marker of drowning, particularly in freshwater. References: [1] Piette, M.H. and De Letter, E.A. (2006), Drowning: still a difficult autopsy diagnosis, Forensic Science International, 163(1-2), 1-9. [2] Azparren, J.E., Ortega, A., Bueno, H. and Andreu, M. (2000), Blood strontium concentration related to the length of the agonal period in seawater drowning cases. Forensic Science International, 108(1), 51-60. [3] Azparren, J.E., Fernandez-Rodriguez, A. and Vallejo, G. (2003), Diagnosing death by drowning in fresh water using blood strontium as an indicator, Forensic Science International, 137(1), 55-59.

147

Firing distance estimation through the analysis of the gunshot residues deposit pattern around de bullet entrance hole

J. Tuna1, A. Santos2, A. Almeida1 and T. Magalhães2

1 REQUIMTE, Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Portugal. 2 Serviço de Medicina Legal, Faculdade de Medicina, Universidade do Porto, Portugal. Serviço de Patologia Forense, Instituto Nacional de Medicina Legal, I.P. – Delegação do Norte.

The determination of the shooting distance is of utmost importance in criminal investigation of firearm incidents since it can decisively contribute for the differential medico-legal diagnosis among a simple accident, a suicide or a homicide [1]. One of the main evidences in these cases is the presence of residues (gunshot residues – GSR) in the skin of the victim, at the impact zone and around it. Such GSR are essentially particles resulting from the condensation of vapors of the “primer” that were jointly transported with the projectile to the impact surface. Regarding metallic elements, GSR mainly contain lead (Pb), antimony (Sb) and barium (Ba). In a forensic ballistics perspective, it is understandable that the quantity (the deposit density) and the distribution pattern of GSR around the bullet entrance hole is somewhat related with the firing distance, a fact that has been explored by some investigators. In a previous work [2], we studied the 6.35-mm pistol, the gun most frequently involved in firearms incidents in Portugal. Test shots were made against a target of cotton tissue, and the amounts of Sb, Ba and Pb deposited in quadrangular pieces (1 x 1 cm) of the target, cut from four radial positions around Fig. 1 – Sampling positions for the bullet entrance hole (Fig. 1), were determined GSR analysis. by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). The data obtained were used to establish a mathematical model for estimating the firing distance. In the experimental conditions used, it was possible to estimate it on the interval [20-80] cm. The best regression curve which adjusted to the data was a linear multiple regression between the firing distance (d) and the logarithm of the mass (m) of each element in samples C (i.e., radial distance between 3.5 cm and 4.5 cm), expressed in µg/g of target tissue: d = a + b1X1 + b2X2 + b3X3, where X1 = ln m (Sb), X2 = ln m (Ba) and X3 = ln m (Pb). The accuracy of firing distance estimation using only 1 or 2 elements was not significantly different from the one obtained with the 3 elements. Using a similar experimental protocol, we have now extended the study to the 7.65 mm pistol, another gun also commonly involved in firearm incidents in Portugal, and the results were quite similar. However, for this pistol, the best radial position for samples collection seems to be position B. References: [1] DiMaio, V.J.M. (1999), Gunshot Wounds: Practical Aspects of Firearms, Ballistics, and Forensic Techniques, CRC Press, Boca Raton. [2] Santos, A., Magalhães. T., Vieira, D.N., Almeida, A.A. and Sousa, A.V. (2007), Firing distance estimation through the analysis of the gunshot residue deposit pattern around the bullet entrance hole by inductively coupled plasma-mass spectrometry: an experimental study, The American Journal of Forensic Medicine and Pathology, 28(1), 24-30.

148

In vitro assessment of whey edible coats with antimicrobial features against differents microbial strains

A. Santos, M. Leão, O. Ramos, M. Pintado, and X. Malcata

Escola Superior de Biotecnologia - Universidade Católica Portuguesa Rua Dr. António Bernardino de Almeida, P-4200-072 Porto, Portugal

A new generation of food packages is on the way – as traditional concepts were hampered in their ability to further extend shelf-life of food products, coupled to considerably high ecological footprints. One element of such a generation takes advantage of coats that inhibit undesirable microbial growth; however, most commercial solutions already made available in the market include compounds – which cannot be ingested. Edible coatings based on milk proteins – viz. whey proteins, have meanwhile been developed for use as protective layers in a variety of foods. Unfortunately, their main functionality relates to barrier properties, so there is large room for improvement in appearance. Development of edible coats exhibiting general antimicrobial properties is thus an issue of practical relevance; however, a general solution may not exist, as different spoilage/pathogenic microflora have been associated to the food surface. Consequently, the objective of this research effort was to assess – via in vitro tests, the efficacy of several formulated antimicrobial edible coats, based on whey protein isolates as base coating material, and glycerol as plasticizer. The antimicrobial activity of several antimicrobial agents — e.g. a polysaccharide (oligochitosan), organic acid (lactic acid) and a bacteriocin (nisin) were tested against yeasts (e.g. Yarrowia lipolytica) and bacteria (Escherichia coli and Listeria innocua) at an inoculum level of 105 cfu/ml. The inhibitory effect of said antimicrobials was initially tested in culture media and in edible coats, which remained in contact with said microbial strains for 24 h. Afterwards, the best antimicrobial systems were selected. Lactic acid (at 0.6%) and oligochitosan (at 2%) yielded the strongest effect under the conditions tested against previously strains. Different coatings showed different antimicrobial behaviors – depending on the microbial agent in stake; however, association of antimicrobial agents increased (as expected) the individual antimicrobial activities.

149

Synthesis, structure elucidation and evaluation of xanthone derivatives for dual activity: antitumor activity and P-glycoprotein inhibition

J. Lima1, A. Paiva1, E. Sousa1,3, M. Pinto1,3, M. S. J. Nascimento2,3 and M. H.Vasconcelos2,4

1 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 Department of Microbiology, Faculty of Pharmacy, University of Porto, Portugal. 3 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal. 4 IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal.

Although enormous progress has been achieved in the field of cancer therapy, only approximately 50 % of all cancers are susceptible to chemotherapy. From these, more than 50 % rapidly develop drug resistance during therapeutical treatment. Most often, this drug resistance is a multiple drug resistance (MDR) phenotype caused by the overexpression of P-glycoprotein (P-gp), a membrane-bound efflux pump which transports a wide variety of anticancer agents out of the tumor cells. Inhibition of P-gp is a powerful approach to reverse MDR; however, current P-gp inhibitors have demonstrated limited clinical success [1]. The development of a library of small molecules that will potentially reveal simultaneous activities, as antitumor and P-gp inhibitor, is in progress, in which the requirements established for P-gp modulation were applied to antitumor xanthones. Using parallel synthesis and solid supported O Cl O NR 2 reagents, several compounds were Cu2O + NHR2 obtained (Fig. 1). MeOH S S r.t. overnight O O CH3 The multiple ligands will be CH3 O O assessed for their antitumor MP-CNBH CHO 3 NR AcOH 2 activity by testing their capacity to

+ NHR2 O OH MeOH O OMe inhibit the in vitro growth of OMe r.t. overnight OH several human tumor cell lines. Fig. (1). Synthesis of amino(tio)xanthones. The in vitro biological evaluation of MDR modulation will be conducted by testing the ability of the molecules to either reduce P-gp protein expression or reduce the efflux of rhodamine from K562Dox cells (cells from a chronic myeloid leukemia cell line with overexpression of P-gp, obtained by another group from K562 cells after prolonged drug [2]). References: [1] McDevitt, C.A. and Callaghan, R. (2007), How can we best use structural information on P- glycoprotein to design inhibitors?, Pharmacology & Therapeutics, 113, 429-441. [2] Lima, R.T., Guimarães, JE and Vasconcelos, M.H. (2007), Overcoming K562Dox resistance to STI571(Gleevec) by downregulation of P-gp expression using siRNAs,Cancer Therapy, 5, 67-76.

Acknowledgments: FCT (I&D, nº226/94), FEDER, POCI, U. Porto, and Caixa Geral de Depósitos for financial support.

150

Synthesis of xanthone derivates for in vitro and in vivo biological activity studies

J. Siroka1,2, E. Sousa2,3 and M. Pinto2,3

1 Department of Organic Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Czech Republic. 2 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 3 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal

Xanthone derivatives are heterocyclic compounds with the dibenzo-γ-pyrone as the main molecular moiety. They contain different types of substituents in different positions, leading to a large variety of pharmacological activities [1]. 3,4-Dihydroxyxanthone (1, Fig. 1) was revealed as a hit compound in a study involving the investigation of the inhibitory effect of oxygenated xanthones on several human tumor cell lines [2]. In order to obtain enough quantity for in vivo assays an for further molecular modifications, the synthesis of 3,4-dihydroxyxanthone (1) was accomplished (Fig. 1): the condensation of 1,2,3-trimethoxyphenol (2) with the appropriate substituted benzoyl chloride 3 afforded Friedel-Crafts acylation O benzophenone 4 which was OMe COCl further cyclized to give 3,4- Et O, AlCl + 2 3 r.t. 22h dimethoxyxanthone (5). OMe OMe OMe 3 OMe OH Compound 5 was demethylated OMe OMe 4 2 to furnish 3,4- MeOH, H2O, NaOH, reflux/47h Base-catalized cyclization dihydroxyxanthone (1). Additionally, the synthesis of O O Demethylation reactive formylated derivatives C H CH , AlCl , 70 ºC/13h 6 5 3 3 of xanthones, 1 and 5, was O OH O OMe 5 attempted by Duff formylation. 1 OH OMe Duff Formylation Only 1-formyl-3,4- HMTA (hexamethylentetramin) dihydroxyxanthone (6) was CF3COOH obtained by this procedure. O CHOHCO O CHO Compound 6 will be used as a building block on the O OH O OMe construction of a library of amine OH OMe 6 xanthonic derivatives to Fig. 1. Synthesis of compounds 1 and 2. investigate antitumor activity.

References: [1] Pinto, M., Sousa, E., Nascimento, M. S. J. (2005), Xanthone derivatives: new insights in biological activities, Current Medicinal Chemistry, 12, 2517-2538. [2] Pedro M., Cerqueira, F., Sousa, M.E., Nacimento, M.S.J. and Pinto, M. (2002), Xanthones as inhibitors of growth of human cancer cell lines and their effects on the proliferation of human lymphocytes in vitro, Bioorganic Medicinal Chemistry, 10 (12), 3725-3730. Acknowledgments: FCT (I&D, nº226/94), FEDER, POCI for financial support.

151

Ultrastructural characterisation of Pirellula sp. OJF20

R. Abreu1, F. Viana1, J. Torres1, J. Bondoso1,2, A. Lobo-da-Cunha2,3 and O.M. Lage1,2

1 Departamento de Botânica, Faculdade de Ciências, Universidade do Porto 2 CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental 3 Instituto de Ciências Biomédicas Abel Salazar

Planctomycetes are a phylum of the domain Bacteria with particular morphologic and structural characteristics, as peptidoglycan-less wall and intracellular compartmentalisation (1, 2). These characteristics make them an important object of study for the clarification of the phylogenetic relationships between microorganisms (the construction of the tree-of- life) as well as prokaryotic and eukaryotic differentiation. Pirellula sp. OJF20 (GenBank accession number-EF589346) was isolated from the surface of macroalgae Corallina sp., sampled in a rocky beach in Foz, Porto, and belongs to a new independent cluster close to the Rhodopirellula genus (94% similarity in the 16S rRNA gene). Electron microscopy techniques such as negative staining, ultrathin section analysis and ultracytochemistry were used to characterise this strain. Results showed that cell organisation and life cycle is similar to that of R. baltica (3). They cells usually present an ovoid shape, although polymorphism can be found especially in older cultures. Ultrathin sectionsl revealed the presence of a paracrystaline structure well evident in the cytoplasm. Probably this structure is a virus but new studies are now being performed to confirm its nature.

1. Fuerst JA. 1995. The planctomycetes: Emerging models for microbial ecology, evolution and cell biology. Microbiology 141: 1493-506 2. Fuerst JA. 2005. Intracellular compartmentation in planctomycetes. Annual Review of Microbiology 59: 299-328 3. Gade, D, Stührmann, T, Reinhardt, R & Rabus, R. 2005. Growth phase dependent regulation of protein composition in Rhodopirellula baltica. Environ. Microbiol. 7: 1074- 84

152

Mesoporous Carbons: Synthesis and Functionalization

C.A. Orge1, J.P.S. Sousa1, F. Gonçalves1, C. Freire2, M.F.R. Pereira1 and J.J.M. Órfão1

1 Laboratório de Catálise e Materiais, Departamento de Engenharia Química, Faculdade de Engenharia, Universidade do Porto, 4200-465 Porto, Portugal. 2 REQUIMTE, Departamento de Química, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal.

Mesoporous carbons have received a great attention due to their potential use as advanced adsorbents and catalytic supports. Comparatively to the traditional microporous activated carbons, the use of these materials in the aforementioned applications presents several advantages, mainly related to its large-pore network, which makes mass transfer limitations less significant. This is of utmost importance when dealing with bulky molecules, such as textile dyes. On the other hand, the surface chemistry of these carbons can be tailored to specific needs, by appropriate chemical and thermal treatments. Several methods can be followed for preparing these materials, such as catalytic activation of carbon precursors, template-based methods, carbonization of polymer/polymer blends and organic cryo-, xero- and aerogels. The present work aimed on the preparation of mesoporous carbons with large surface area, high porosity and a controlled, narrow pore size distribution. Two main series of materials were produced: a) carbon xerogels (CX), prepared by sol-gel condensation of resorcinol and formaldehyde [1]; and b) templated carbons a) (CMK-3), using a previously synthesised silica 0.15 CX CX-HNO

) 3 (SBA-15) as template [2]. After the synthesis step, -1 CX-O

.g 2 the surface chemistry of the carbons prepared was -1 0.10 mol.s μ

modified by means of gas and liquid phase ( 2 0.05 treatments, with O2 and HNO3, respectively. The CO nature and amount of the surface oxygen groups 0.00 introduced by the different treatments was then 02505007501000 Temperaure (ºC) analysed by temperature-programmed desorption b) 0.9 (TPD) and the textural characterization of all the ) -1

.g 0.6 carbons prepared was based on the N2 adsorption -1 isotherms at -196 ºC. Figure 1 shows the TPD mol.s μ profiles obtained for the CX samples. 0.3 CO ( In a second phase of the work, the synthesised 0.0 carbons will be used in adsorption processes and in 02505007501000 the preparation of oxidation catalysts (e.g. anchored Temperaure (ºC) metalloporphyrins) for the treatment of textile Figure 1 – TPD spectra for the CX. effluents. a) CO2 evolution; b) CO evolution. Acknowledgements: This work was partially funded by the program Investigação Científica na Pré-Graduação 2007, (Universidade do Porto and Caixa Geral de Depósitos - project IPG58) and by Fundação para a Ciência e a Tecnologia (POCTI/1181). FG thanks Fundação para a Ciência e a Tecnologia for the project POCI/N001/2005. References: [1] Mahata, N., Silva, A.R., Pereira, M.F.R., Freire, C., Castro, B., Figueiredo, J.L. (2007), Anchoring of a [Mn(salen)Cl] complex onto mesoporous carbon xerogels, J Colloid Interf Sci 311 152-158. [2] Fuertes, A.B. (2004), Synthesis of ordered nanoporous carbons of tunable mesopore size by templating SBA-15 silica materials, Micropor Mesopor Mat 67 273-281.

153

Physical exercise and stress hormones

M. Moz1, S. Baptista1, O. Cardoso1, M. Mendanha1, A. Ascensão2, J. Magalhães2 and L. Ribeiro1

1 Department of Biochemistry, Faculty of Medicine, University of Porto, Portugal. 2 Department of Sport Biology, FCDEF, University of Porto, Portugal.

An adequate balance between catabolic processes (mobilization of energy), induced by stress hormones such as the catecholamines (CA), adrenaline (AD) and noradrenaline (NA) and cortisol, and anabolic processes (repair, healing, growth), induced by steroid sex (such as testosterone) and growth hormones, is vital for health and survival [1]. Regular moderate physical exercise is known to contribute to such anabolic effects and to an earlier homeostasis restoration [2]. However, long-term and/or excessive physical exercise result in the over-secretion of CA and cortisol, leading to over-reaction and exhaustion of the target-organs, which can be extremely harmful. Acute or chronic stress, induced by excessive physical exercise, may thus lead to an inadequate endocrine response, eventually contributing as a risk factor to disease initiation and/or exacerbation [3]. The aim of our work was to examine the effect of exposure to an intense and extremely stressful training on several plasma stress hormone levels in healthy male individuals. Two groups were evaluated: one group of individuals conditioned by a previous 3 months rigorous training (Group A; n=7) and another one not submitted to this training (Group B; n=7). Blood samples were collected before (basal), immediately after 72 hours of the training and 48 hours (rest period) after this period. AD and NA levels were quantified by means of high pressure liquid chromatography with electrochemical detection (HPLC-ED) and plasma cortisol and testosterone levels by RIA.. There were no significant differences between groups A and B, relatively to basal levels, of either cortisol (13.4±1.64 vs. 14.7±1.1 µg/dL) or testosterone (136.6±40 vs. 176.7±40 ng/dL). After training, there was an increase of cortisol plasmatic levels, which was though only significant for group A. However, after 48 hours of rest, cortisol levels remained high in group B, but not in A (p<0.05). In both groups, there was a significant increase of testosterone plasma levels after the 48 h period of rest. There was no significant differences in AD levels between groups A and B either under basal, after exercise or after post-exercise rest. As to NA levels, group B showed higher basal values than group A. However, they have been similarly decreased by exercise and after post-exercise rest. In conclusion, our preliminary results suggest that in order to adequately respond to an intense and extremely stressful exercise it is necessary a good fitness condition. In the near future, other stress parameters will be evaluated.

[1] Tsigos, C. and Chrousos, G.P. (2002) Hypothalamic-pituitary-adrenal axis, neuroendocrine factors and stress. J Psychosom Res 53: 865-71. [2] Lakka, T.A. and Laaksonen, D.E. (2007) Physical activity in prevention and treatment of the metabolic syndrome. Appl Physiol Nutr Metab 32: 76-88. [3] Lundberg, U. (2005) Stress hormones in health and illness: the roles of work and gender. Psychoneuroendocrinology 30: 1017-21.

154

Single-standard calibration applied to SIA analytical system with potentiometric detection

Leonel G. C. Soares1 and Rui A.S. Lapa1

1 REQUIMTE/CEQUP, Faculty of Pharmacy, University of Porto, Portugal.

The flow techniques provide a wide range of possibilities being calibration and dilution schemes applicable to a variety of analytical methodologies [l, 2]. Among the flow techniques, the Sequential Injection Analysis (SIA) analytical systems have been applied in the refinement of calibration techniques to accompany the wide range of analytical applications. Different calibration methods has been proposed [3,4] and could be classified as: (a) the conventional method (CM), (b) the indirect method (IM), (c) the internal standard method (ISM) and (d) the dilution method (DM) [5]. The conventional calibration method is commonly used in analytical practice in two modes: interpolative and extrapolative, which are usually termed ‘the calibration curve method’ and ‘the standard addition method’, respectively [5]. In the first one, the calibration graph is prepared with the use of a set of standard solutions prepared separately from a sample; as a consequence, the signal measured for the sample is then related to the calibration graph and the analytical results are interpolated. In the ‘the standard addition method’ standard solutions are added to the sample, hence the calibration graph is limited by the signal obtained for the sample and the analytical result can be evaluated exclusively by extrapolation. The advantages of the extrapolative calibration process reside in the compensation of the effect of the interferent species since the interpolative method presents the effect of those interferences. In the work, a simple extrapolative process applied to a SIA technique was developed to generate a set of precisely diluted aliquots from a single standard and applied to the determination of chloride in water sampled. An ion selective electrode sensitive to chloride was used as detector.

References: [1] - Ruzicka, J., and Hansen, E. H., Flow Injection Analysis, Wiley, New York, 2nd ed., 1988. [2] - Ruzicka, J., Anal. Chem., 1983,55,1986. [3] P. Koscielniak, Anal. Chim. Acta, 438 (2001) 323. [4] P. Koscielniak, in: J. Niemiesnik, W. Chrzanowski (Eds.), New Horizons and Challenges in Environmental Analysis and Monitoring, Centre of Excellence in Environmental Analysis and Monitoring, Gdansk, 2003, 110. [5] - Paweł Koscielniak, Marcin Wieczorek, Joanna Kozak, Małgorzata Herman, Analytica Chimica Acta 600, 2007, 6.

155

PHOTOCATALYTIC DEGRADATION OF CIPROFLOXACIN ANTIBIOTIC IN TiO2 AQUEOUS SUSPENSION

F. G. Azevedo, M. Domingos, C. G. Silva and J. L. Faria

Laboratório de Catálise e Materiais, Departamento de Engenharia Química, Faculdade de Engenharia, Universidade do Porto, Rua Dr. Roberto Frias 4200-465 Porto, Portugal

The presence of pharmaceuticals and their metabolites in aquatic environments has raised increasing concern in recent years [1]. Antibiotics can hardly be degraded by biological processes, leading to the persistence of these compounds in the aquatic ecosystem posing a real threat in what concerns bio-accumulation. Hence, it is a priority to search for alternative chemical treatment methods, which can effectively eliminate the disposed amounts of this type of compounds. Ciprofloxacin (CIPRO) is an antibacterial drug belonging to the class of fluoroquinolones, widely used in the treatment of severe infections in humans. Among the possible treatments, TiO2-mediated photocatalysis has proven to be very effective in the complete oxidation of a great variety of organic pollutants [2]. The process is based in the oxidative potential of the hydroxyl radical (HO•), which reacts rapidly and non-selectively with a wide range of organic compounds. In this work, it is shown that CIPRO can be degraded in an aqueous suspension of TiO2 (Degussa P-25) using ultraviolet (253.7 1.0 nm) radiation, more efficiently that in the absence of any photocatalyst (Fig 1). The photocatalytic process leads to a total 0.8 organic carbon removal of 80% after two hours of irradiation. 0.6 UV

The aim of this study is to examine the mechanism of photocatalytic reaction. C/C0 0.4 The effects on the reaction of the TiO2 catalyst loading, CIPRO initial 0.2 UV/TiO2 concentration (C0), pH, irradiation intensity and dissolved oxygen 0.0 concentration are described. A kinetic 0102030405060708090100110120 Time (min) model to describe the contribution of the mentioned variables will be attempted. Fig. 1- CIPRO decay during photochemical (UV) and photocatalytic (UV/TiO2) reactions. Acknowledgments: The authors gratefully acknowledge FCT and FEDER for financial assistance (fellowship SFRH/BD/16966/2004 and projects POCI/EQU/58252/2004 and POCTI/1181/2003).

References: [1] Hirsch, R., Ternes, T., Haberer, K. and Kratz, K.-L. (1999), Occurrence of Antibiotics in the Aquatic Environment, The Science of the Total Environment, 225(1-2), 109-118. [2] Silva, C.G., Wang, W. and Faria, J.L. (2006), Photocatalytic and Photochemical Degradation of Mono-, Di- and Tri-azo Dyes in Aqueous Solution Under UV Irradiation, Journal of Photochemistry and Photobiology A: Chemistry, 181(2-3), 314-324.

156

Anti-Angiogenic Effects of Ranibizumab and Bevacizumab in Age-Related Macular Degeneration: Effects on Human Endothelial Cells

M. Ferreira-Pinto1*, T. Taveira-Gomes1*, A Carneiro2, M Falcão2, I. Azevedo1, F. Falcão-Reis2, R. Soares 1

1 Department of Biochemistry, Faculty of Medicine of University of Porto, Portugal. 2Department od Ophthalmology, Faculty of Medicina of University of Porto, São João Hospital, Portugal * The two authors contributed equally to the project

INTRODUCTION: Age-related macular degeneration (AMD) is the leading cause of irreversible blindness over 50 years of age in developed countries. This pathology is characterized by the development of abnormal choroidal blood vessels that proliferate through the Bruch's membrane, invading the subretinal space, beneath the macula, which causes severe and fast loss of vision. Angiogenesis, the formation of new blood vessels from pre-existing ones, is accomplished by a huge number of pro-angiogenic growth factors, such as vascular endothelial growth factor (VEGF). Recent reports claiming that VEGF plays a crucial role in AMD-related angiogenesis rendered this growth factor a major therapeutic target. Ranibizumab and bevacizumab are two therapeutic agents already being used in the clinical pratice that block the action of VEGF. However, there are no studies comparing the relative efficacy and safety in AMD patients.

OBJECTIVES: The purpose of the present study is to compare the effects of ranibizumab and bevacizumab in endothelial cell growth, apoptosis, migration and in vitro capilary-like tubule formation.

METHODS: Human Umbilical Vein Endothelial Cell (HUVEC) cultures were incubated with different concentrations of ranibizumab or bevacizumab within the doses used in the clinic, or their excipients. HUVEC viability (by MTT), proliferation (by BrdU immunoassay), apoptosis (by TUNEL assay) and migration (using double-chamber assays) were performed. Evaluation of tubule-like structures formation was performed on matrigel- coated plaques incubated with ranibizumab, bevacizumab or their excipients.

RESULTS: Neither bevacizumab nor ranibizumab presented cytotoxic effects, as evaluated by MTT assay. HUVEC proliferation was significantly reduced by the two agents, as compared to excipient-treated controls. Incubation with bevacizumab at five different concentrations led to increase apoptosis. Ranibizumab treatment resulted in increased apoptosis in the two higher concentrations tested. Cell migration was only affected by higher concentrations of both agents. Cell assembly into capillary-like structures was effectively downregulated by incubation with bevacizumab at the clinically used dose, whereas ranibizumab treatment resulted in decreased cord formation, although not reaching statistical significance.

CONCLUSIONS: This study demonstrated that clinical doses of bevacizumab and ranibizumab are able to prevent several steps of the angiogenic process in a distinct manner. Statistical analyses are being performed in order to elucidate the precise effects of each of the agents examined.

157

Geometry and Space Conception the importance of the processes of representation to Architecture

Bruno Russo1 and João Pedro Xavier2

1 Faculty of Architecture, University of Porto, Portugal. 2 Doctor, Professor of Geometry, Faculty of Architecture, University of Porto, Portugal.

What is the nature of space? What is our concept of space? Has it been unchangeable over time? And how do the architects use this concept, or concepts, in order to create architecture, the organization of space? The search for the idea of space starts at the crossroads of Geometry and Architecture. If the latter is its organization and management, the first, more than the science that studies space, is its very language, the ultimate tool for the architect, even if sometimes he’s not aware of it. Geometry and Architecture have evolved through time and their paths have often crossed, causing echoes on the individual development of each. Starting at Euclid’s Elements, the cornerstone of the western knowledge on geometry, and following the major breakthroughs of this science of space, we try to understand the influence they set on architectural production. On the other hand, by identifying turning points on Architecture’s theory and/or practice we’ll search for reverberations on the representation processes in particular, and on Geometry in general. Architecture owes Geometry what Vitruvius called dispositio – the representation processes. And since the ancient Greeks those processes have evolved: Euclid thought of an infinite and plane space (Euclidean Space), Brunelleschi, Alberti and Piero della Francesca invented the Perspectiva Artificialis, Desargues and Poncelet developed the Projective Geometry, Monge codified a double-projection system, Lobachevsky, Bolyai and others discovered the non-euclidean geometries, used by Einstein to re-design our model of the Universe and finally, at the turn of the millennium, the computer-based design knows mass use and becomes the only tool for many young architects. The architects used the tools Geometry provided them, and “tuned” them. We cannot sever perspective or Monge’s system from Architecture, and certainly can’t dissociate the classical orders from Pythagoras’ view of the World. Nowadays we get entangled in an overwhelming variety of forms and geometries, in a lack of “order” for the architect to follow. Architecture pursued geometric values on the search of beauty and order for many centuries, but now it seems these values have changed. Or have they not? We think that the relationship between Architecture and Geometry is a symbiosis that can’t be easily broken. We believe that the study of the history of Geometry can help us understand the history of Architecture. Today we have more knowledge, more technology, advanced engineering and higher levels of education. We need to know better the tools at our disposal in order to produce better architecture. Geometry has always been our premium tool but now, with the automation of the processes brought to light by the personal computer, we tend to distance ourselves from the geometric definition of space and form. And this can lead to a poorer comprehension of space, preventing Architecture from achieving higher goals.

158

Wet Peroxide Oxidation and Wet Oxidation of Nitrophenols in Aqueous Streams

S. Martins1,2, A. Ribeiro2, A.M.T. Silva1, P. Araújo2, J.L. Figueiredo1, and J.L. Faria1

1 Laboratório de Catálise e Materiais, Departamento de Engenharia Química, Faculdade de Engenharia da Universidade do Porto, Rua Dr. Roberto Frias s/n, 4200-465 Porto, Portugal 2 CUF – Químicos de Portugal, S.A., Quinta da Indústria, 3860-680 Estarreja, Portugal

The treatment of nitrophenols, typically found in the effluents from the process of nitrobenzene synthesis, was carried out by wet oxidation (WO) and wet peroxide oxidation (WPO). These processes are based in the principles of wet air oxidation, an efficient process for treatment of high strength effluents which cannot be treated by conventional methods [1]. In this process the organics are oxidized in the liquid phase at temperatures around 300ºC and pressures of 5 to 200 bar, in presence of an oxygen-containing agent (usually air). This process has low operating costs and minimal air pollution discharges, the main limitations being the safety implications 1,6 associated with a system operating at such DNP conditions [2]. Under the usual conditions, the 1,2 TNP organic compounds are mainly oxidized into 0,8 C/C0 carbon dioxide and water. Our previous studies 0,4 show that this process is efficient above 200ºC 0 and 7.0 bar of O2 in the degradation of low -30 -15 0 15 30 45 60 75 90 105 120 dinitrophenol (DNP) and trinitrophenol (TNP) t (min) concentrations (below 0.4 g/L) [3]. However, Figure 1 – Evolution of the C/C0 ratio to these operating conditions are quite severe for DNP and TNP. industrial scale-up and higher concentrations of nitrophenols (above 10 g/L) are usually found in real effluents. Therefore, the use hydrogen peroxide (H2O2) as additional oxidizing agent was investigated in this work (wet peroxide oxidation process) aiming to obtain a high efficiency at lower temperatures and pressures. In a wide range of concentrations of nitrophenols the use of mild conditions for temperature and O2 pressure (without H2O2) resulted in conversions up to 99.3 and 98.5% for DNP and TNP, respectively (Fig.1). An initial period of 30 min was necessary to achieve the working temperature. At the same temperature, with an oxygen pressure of 0.8 bar with the higher peroxide concentration used, a complete destruction of nitrophenols was observed. Reducing temperature, keeping the oxygen pressure, but reducing peroxide concentration to 56.1 g/L, resulted in maximum conversions of 92.1 and 99.8% for DNP and TNP, respectively. Global TOC removal efficiencies ranged from 57 to 97%. Therefore, the WPO is strongly recommended in the degradation of nitrophenols. References: [1] – Mishra, V.S, Mahajani, V.V. and Joshi J.B. (1995) Wet Air Oxidation, Industrial & Engineering Chemistry Resaerch 34(1), 2-48. [2] – Kolaczkowski, S. T., P. Plucinski, et al. (1999). Wet air oxidation: a review of process technologies and aspects in reactor design, Chemical Engineering Journal 73(2), 143-160. [3] – Apolinário, Â .C., Silva, A.M.T., Machado, B.F., Gomes, H.T., Araújo, P.P., Figueiredo, J.L., Faria, J.L. (2007), Wet air oxidation of nitro-aromatic compounds: reactivity on single- and multi component systems and surface chemistry studies with a carbon xerogel, Applied Catalysis B, Environmental, (in press, doi:10.1016/j.apcatb.2007.12.018).

159

Formation of heterocyclic aromatic amines in charcoal grilled sardines

M. Costa1, A. Melo1, C. Petisca2, I.M.P.L.V.O. Ferreira1, O. Pinho1,2

1 REQUIMTE- Serviço de Bromatologia, Faculdade de Farmácia da Universidade do Porto, Rua Anibal Cunha 164, 4099-030 Porto; Portugal 2 Faculdade de Ciências da Nutrição e Alimentação, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto – Portugal

Heterocyclic Aromatic Amines (HAs) are produced during cooking at high temperatures of proteinaceous food such as meat and fish [1]. HAs have proved to be carcinogenic in animals and to induce early lesions in the development of cancer [1-2]. Grilling is considered a healthy method for cooking meat and fish, because foods do not sear in a lot of fats. Grilled sardine Sardina pilchardus (Walbaum) is very appreciated by Portuguese people, specially, during summer season. However, high temperatures can be used during cooking and no studies were found concerning HAs’ formation. In the present study, the effects of grilling on the formation of HAs of charcoal grilled sardines were investigated. Sardines were grilled in charcoal, under ordinary conditions (250-280 ºC) at two different degrees of doness, “rare” and “well done”. Extraction and purification of HAs was performed according to the method developed by Gross and Galceran [1-2] and adapted by our research team [3]. The method uses the coupling of LLE with diatomaceous earth as solid support and two SPE steps with propylsulfonic acid (PRS) and C18 was used to extract HAs polar and less-polar HAs. Separation and quantification of HAs was performed by liquid chromatography with diode array and fluorescence detection. All the analytes were quantified by a standard addition method, using two fortified levels (around 5-20 ng g-1) and two non-spiked samples. No HAs were quantified in “rare” grilled sardines, however, seven HAs, namely IQ, MeIQx, PhIP, Trp-P-1, Trp-P-2, AαC, MeAαC were encountered in “well done” grilled sardines. Nonpolar HAs (Trp-P2, AαC, MeαAC) and PhIP were the most abundant HAs in “well done” grilled sardines. The presence of nonpolar HAs, Trp-P-1, Trp-P-2, MeαAC and AαC is not often reported. Chemical modelling has shown that these HAs are generated from pyrolysis of aminoacids via free radical reaction. These compounds have been thought to be formed exclusively under extreme cooking conditions. Intake of sardines is recommended owing to its high level of ω3 content, however, care should be taken concerning the degree of doness.

References: [1] Gross, G.A. (1990) Carcinogenesis (11) 1597- 1603.

[2] Galceran, M.T., Pais, P. and Puignou, L. (1996). Journal of Chromatography A (719) 203-212.

[3] Melo, A., Viegas, O., Eça, R., Petisca, C., Pinho O. and Ferreira I.M.P.L.V.O. (2008), Journal of Liquid Chromatography & Related Technologies (31) 1–16.

160

Influence of beer antioxidative properties on the formation of the heterocyclic amine PhIP in grilled beef

A. Melo1,M. Costa1, R. Eça1, I.M.P.L.V.O. Ferreira1, O. Pinho1,2

1 REQUIMTE- Serviço de Bromatologia, Faculdade de Farmácia da Universidade do Porto, Rua Anibal Cunha 164, 4099-030 Porto; Portugal 2 Faculdade de Ciências da Nutrição e Alimentação, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto – Portugal

PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine) is an heterocyclic aromatic amine belonging to a class of mutagens found in food. It is formed during the heating of food (meat, fish) that contains creatine, free amino acids and carbohydrates. PhIP was first isolated and identified from fried ground beef by Felton [1]. In several publications it was stated that the presence of antioxidants reduces the content of heterocyclic aromatic amines, including PhIP, in meat. Since radical reactions play an important role in the formation of heterocyclic aromatic amines, it was hypothesised that antioxidants should reduce the content of these mutagenic substances in grilled meat. Consequently, the addition of ingredients rich in antioxidants is considered a promising measure to reduce heterocyclic aromatic amines exposure [2-4]. The aim of this study was to investigate the influence of beer, which is expected to have antioxidative properties, on the occurrence of PhIP in grilled beef, one of the most common heterocyclic aromatic amines found in grilled meat. Beef samples were grilled in a teflon-coated frying pan, without adding oil. Before grilling, samples were marinated with beer during 1 h, 2h, 4h and 6h. Control beef samples were not marinated. The heat source was a gas cooker, and the temperature, on the surface, ranged from 180 to 200ºC. Extraction and purification of PhIP was performed by coupling of LLE with diatomaceous earth as solid support and two SPE steps with propylsulfonic acid (PRS) and C18. Separation and quantification of PhIP was performed by liquid chromatography with diode array and fluorescence detection. Fluorescence detector was set at excitation 307 nm and emission. The marination time affected the PhIP formation. Higher content of PhIP was found in control beef samples. There was an effective decrease of PhIP levels along the marination with beer. Further work has to be done to clarify the complex reaction mechanisms for the formation of PhIP. A reduction of the daily human exposure to this and other heterocyclic aromatic amines must be the primary aim. Consequently, changes in cooking habits are needed. References: [1] Felton, J.S., Knize, M.G., Shen, N.H., Lewis, P.R., Andresen, B.D., and Happe J. (1986), Carcinogenesis (7), 1081–1086. [2] Vitaglione, P. and Fogliano, V. (2004), Journal of Chromatography B (802), 189-199. [3] Lan, C.M., Kao, T.H. and Chen, B.H. (2004), Journal of Chromatography B (802), 27-37. [4] Murkovic, M., Steinberger, D., Pfannhauser, W. (1998), European Food Research and Technology (207), 477-480.

161

THE STUDY OF DE FÂNZERES-GONDOMAR GRANITES: A MULTIDISCIPLINARY APROACH

M. Alves1

1 Dep.Geologia, Fac. Ciências, Univ. Porto, Rua do Campo Alegre, 687 4169-007 Porto, Portugal ([email protected]).

The granites from Fânzeres-Gondomar area («granito gnaissico de Fânzeres» and «granito de Gondomar») are located in the Central Iberian Zone. Multidisciplinary studies, namely Anisotropy of Magnetic Susceptibility (AMS), geometry of fluid inclusions plans (FIPs) studies and microthermometric fluid characterization, were carried out. The Gondomar granite presents an average magnetic susceptibility of 29.06 µSI, AMS indicates a magnetic foliation (N19º; 69ºSE) which is the result of the magmatic flow (magmatic foliation). The most representative FIPs trending NNE-SSW, are parallel to the magnetic foliation, which indicate that they enhanced a pre-existent magmatic anisotropy. The fluids that circulate within these micro-cracks are aqueous with high salinities (19.9 to 26.4 mass% eq. NaCl) and minimum temperature of entrapment between 90 and 120ºC. This type of fluids has been referred to as being a result of a contribution of meteoric fluids from late to post-Variscan cover. The Fânzeres granite presents a gneissic foliation, which is visible on the field and on the microscope. The average magnetic susceptibility is 40.97 µSI and the magnetic foliation (N26º; 58ºSE) is parallel to the gneissic foliation. FIPs trending NW-SE are the most representative. They are characterized by aqueous fluids of low salinity (0.2 to 1.2 mass% eq. NaCl) and minimum temperature of entrapment between 130 and 200ºC. These FIPs are perpendicular to the gneissic and magnetic foliation, which agrees with the process of the formation of FIPs. We assume that the FIPs correspond to a stress field where the minimum stress, σ3, is normal to the FIPs that materialize the vertical plane σ1 σ2.

Keywords: granites, anisotropy of magnetic susceptibility, fluid inclusions planes.

162

Educational Sciences and Medical Education: An internship in FCM-UNL Department of Medical Education

R. Encarnação 1

1 Faculty of Psychology and Educational Sciences, University of Porto, Portugal.

This paper describes a project developed during a curricular internship (integrated into the last year of the basic degree in Educational Sciences, held by FPCEUP - Faculty of Psychology and Educational Sciences of Porto University) which took place in the Department of Medical Education (DEM) of the New University of Lisbon’s (NUL) Faculty of Medical Sciences (FMSc). This pre-graduate internship was developed with the support of a grant derived from the POCI2010 programme, from January to June 2006, under the supervision of Professors Rui Trindade (remotely, at FPCEUP) and Patrícia Rosado Pinto (locally, at DEM). We will focus on ways in which this work can be characterized as a potential field of action for an Educational Sciences professional, in a Higher Education-specific context like Medical Education. One of the main objectives of this internship was to gain a deeper understanding of the Department’s body of work and its role within the Faculty of Medical Sciences, taking into account the different topics studied during our pre-graduate years, specifically focusing on if and how the DEM’s role could be characterized as Educational Assessment and Mediation. We believe that the DEM can be seen as a point upon which the Educational and Medical Sciences can converge, in a cross-fertilization, multi-focal and reflexive approach. A full-immersion approach was outlined in order to accomplish these objectives, having been stated since the initial project that the intern’s role would be that of a normal element of the DEM, as a way to experience first-hand how the Department functioned and worked. Weekly activity reports were instated as mandatory, in order to collect information and thoughts on the intern’s, and the Department’s activities and characteristics. We will present the work developed at the Department of Medical Education during the internship, analyzing its activity in light of the Educational Sciences’ rationale of intervention in education.

163

Quartz Crystal Microbalance: A Useful Tool As Sensor Of Thin Films

J. Borges, C. M. Pereira and F. Silva

Department of Chemistry, Faculty of Science, University of Porto, Portugal. Rua do Campo Alegre, 687, 4169-007 Porto, Portugal

The quartz crystal microbalance (QCM) is a simple, cost effective, high-resolution mass sensing technique, based upon the piezoelectric effect [1]. A quartz crystal microbalance (QCM) is an electromechanical transducer that can produce an electrical signal in response to inertial and viscoelastic changes of a quartz crystal when a voltage is applied across its faces [2]. The QCM then measures the frequency of oscillation of the crystal and possesses a wide detection range. The QCM can detect changes in frequency of the crystal due to changes in mass deposited on the surface of the crystal. This characteristic contributes for the development of sensors [3]. Figure 1 shows a typical result obtained in the study on the deposition of Oleylamine by adsorption at the surface of gold plated quartz crystal using the QCM-Z500 instrument.

400 ∆f = f (t) 350 300 Fundamental 250 3rd Overtone 200 5th Overtone 150 7th Overtone 100 9th Overtone Figure 1. Changes in ∆f 50 11th Overtone 0 due to the deposition by -50 05001000150020002500300035004000 adsorption of Oleylamine -100 at the surface of the gold -150 plated quartz crystal.

Resonant frequency shift / Hz / shift frequency Resonant -200 -250 -300

time / sec

The experimental results will be analyzed in order to characterize the adsorption behaviour of Oleylamine at gold surfaces and results will be presented regarding the thermodynamics of the adsorption process.

Acknowledgments: Financial support from FCT (POCI 2010) and FEDER for the project POCI/QUI/57679/2004 is gratefully acknowledged. J. Borges acknowledges the project POCI/QUI/57679/2004 for a BI grant. References: [1] Kenneth, A. M., Biomacromolecules, 2003, 4 (5), 1099-1120. [2] Etchenique, R. A.; Calvo, E. J., Anal. Chem., 1997, 69, 4833-4841. [3] Pei, Z.; Anderson, H.; Aastrup, T.; Ramström, O., Biosensors and Bioelectronics, 2005, 21, 60- 66.

164

João Andresen’s Fifties Houses: Portuguese Domestic Architecture Up-to-Date

Sergio Dias da Silva1

1 Architecture Graduate, Faculty of Architecture, University of Porto, Portugal.

João Andresen, born in Porto in 1920, became an Architect by the School of Fine Arts in 1948, beginning in that same year both his professional and academic careers, which would last until his death in 1967. His work encompassed several fields, as Urbanism, Housing, Public Buildings, Monuments and Furniture Design. This study1 of Andresen’s work is a consequence of the consultation and analysis of his estate and of specialized periodic publications of the time, as of the visit to some of his buildings. The investigation was complemented by the analysis of some bibliography Andresen recommended to his clients - “The Culture of Cities” (Mumford, L., 1938), “Mechanization Takes Command” (Giedion, S., 1948) and “Post-War Architecture: Built in USA” (Hitchock, H.-R., Drexler, A., 1952). In 1954, Andresen entered the International Calvert House Competition for the Canadian Home of Tomorrow, a first for Portuguese architects on international competitions2; for that purpose, he created a “Future House”, symbol of his ideas of modern ideal home and on the future of architecture. Between 1953 and 1956, some of Andresen’s houses represent similar ideas to the “Future House”. The House in Caxias (1953-55) and House in Figueira da Foz (1956-58) are visually similar to the “Future House”, but the first use of those principles was in the House in Valongo, of 1953, usually considered closer to Brazilian modern architecture3. Andresen’s role, and of his generation, in the questioning of the Modern Movement is yet to be fully perceived, as his colleague, Fernando Távora (1923-2005), became the undisputed leader of a new time in Portuguese architecture. These Andresen’s houses, inspired in the binuclear houses of the architect Marcel Breuer (1902-1981), confirm that the American architecture is the reference to a new way of designing domestic space4 but show, simultaneously, an abstract understanding of tradition that was to become, after all, the main legacy of the Enquiry to Portuguese Popular Architecture (1955-61). The study of Andresen’s houses of the fifties, and of other Portuguese architects’ single- family houses of the same decade, shows a generation’s ability to keep up to the main researches in the design of domestic space, never abandoning peculiar aspects of the Portuguese context and architecture. This generates an adaptation of international models, a trademark of Portuguese architecture, and a unique view on the reinterpretation of the Modern Movement. [1] Silva, S. (2007), João Andresen: Uma Ideia de Arquitectura, Graduation Thesis, FAUP, Porto [2] Toussaint, M. (2003), De Dentro para Fora na Década de 50, Jornal Arquitectos nº212 [3] Fernandez, S. (1985), Percurso – Arquitectura Portuguesa 1930-1974, FAUP, Porto, p.102 [4] Colomina, B. (1997), “Built in the USA: La Casa Americana de Posguerra”, in La habitación y la ciudad modernas : rupturas y continuidade : 1925-1965 : actas / Primer Seminario Docomomo Ibérico, Zaragoza, p.197

165

Synthesis of xanthone derivatives by microwave-assisted methods

P. Vrbata1, M. Pinto2,3, R. Castanheiro2,3, S. Cravo2,3

1 Department of Organic Chemistry, Faculty of Pharmacy, Charles University, Prague, Czech Republic. 2 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 3 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal.

Xanthone derivatives are very interesting compounds and our research group has a large experience in synthesis of these bioactive derivatives by classic ways, generally with harsh reaction conditions [1,2]. For that reason, it was performed reactions with microwave (MW) irradiation or MW combined with adsorbed reactants onto a solid support such as clays, in order to obtain new xanthone derivatives in a more environmentally friendly manner, by performing higher-efficiency reactions using fewer and/or safer reagents. So, in this work is described the synthesis of xanthonic building blocks 1 and 2 with MW irradiation (Fig.1) and the dihydropyranoxanthones 3 and 4 (Fig.1) through a combined method of MW with heterogeneous catalysis. The structural elucidation of the compounds will be established by spectroscopic methods, mass spectrometry, and additionally by X-ray crystallography. The compounds 2-4 will be evaluated for biological activities, namely antitumor activity in human cancer cell lines.

Fig.1. Synthesized xanthonic derivatives.

References: [1] Pinto, M.M.M., Sousa, M.E. and Nascimento, M.S.J. (2005), Xanthone Derivatives: New Insights in Biological Activity, Current Medicinal Chemistry, 12 (21), 2517-2538. [2] Pinto, M.M.M. and Sousa, M.E. (2005), Synthesis of Xanthones: An Overview, Current Medicinal Chemistry, 12 (21), 2447-2479. Acknowledgments: FCT (I&D, nº226/94), FEDER, POCI, U. Porto, and Caixa Geral de Depósitos for financial support.

166

Location and Ownership of the Natural Resources in Iraq

C. Silva ; H. Azevedo

¹Department of Geography, Faculty of Letras, University of Porto, Portugal

The Geography, with its traditional emphasis in demarcating, measuring, describing and classifying the territory, is naturally important for the politics. Nowadays there is a new geography and a new typology of the conflicts, where the priorities of Countries are related with the natural resources. The geopolitical, economical and ecological perspectives are the main causes of the "wars of resources", in other words (Klare 2001), they are responsible for the "conflicts that happen for the ownership of critical materials." This term also includes the "description of the fights of local populations against the resource exploration and the neoliberal reforms in control of the resources" (Gedicks 1993; Perreault 2006). Many people cannot be sustained properly in the world, because their countries were not formed naturally. They are frequently dominated by external powers, generally the Western countries. The western geopolitical thinking on resources was dominated by the trade, war and power equation. Nowadays, we attend to the dispute of territories for the most important and valued resources, namely the oil-producing products, by the designated developed western society. The main cause of the importance of the resources has to do with their shortage, with the need of the consumer’s satisfaction and with the implications in the world distribution. With the elaboration of this work for class “Geopolitics and Natural Resources”, it was our objective to deepen our knowledge concerning the most valued natural resources to the world scale, of the Middle East and of Iraq, realizing if these have evident influence in the per capita income of the population and in the actual situation that the World and the country crosses. For such, we proceeded to the collect of available statistical information with the intention of proceeding to its graphic and cartographic representation. In this extent, we collected data concerning the population, per capita income and natural resources. The scientific procedure adopted was the deductive, as the project is structured in three scales: world; regional (Middle East) and local (Iraq). The actual energy needs of the world economy are satisfied, essentially, by non-renewable energies, as the oil, natural gas and coal. The developed economies, as the Westerner, are energetically dependent of countries, like Iraq, in that its economy is extremely dependent of the exports of oil. Iraq possesses natural resources that guarantee the energy independence. However, it continues to have social problems, due to the sanctions that are imposed because of the frequent conflicts. As Iraq has to import the goods essential to the survival of the population, the diversification of its economy would be fundamental, betting more in the primary and tertiary sectors, in a way to contribute for the economical growth of the country, reducing the external dependence and supplying job to the population, turning the country more competitive globally. If Iraq cuts in the provisioning of oil, it will cause serious economical crisis in the countries that are dependent of this resource. We are in the presence of a closed world political system where all the crisis of an exportation area would be reflected all over the world, in all the regions, even if geographically distant.

167

Synthesis, Structural Elucidation and Biological Activities of Chiral 1-Amino-2-Propanol Xanthone Derivatives

I. Teixeira1, M. Pinto1,2 and C. Fernandes1,2

1 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal.

There is a large variety of natural and synthetic xanthone derivatives described in the literature [1] but only a few examples with a chiral moiety [2]. Previous research has shown that chiral xanthones, namely chiral aminoalkanolic derivatives, present a sort of pharmacological activities, such as anticonvulsant and antidepressant activities [2]. Our research group (CEQOFFUP) has a large experience in synthesis of xanthones with interesting biological activities, namely among then a chiral xanthone obtained by the condensation of 2-carboxy-6-methoxyxanthone with the aminoalcohol L-valinol. In this presentation it is shown the synthesis of new chiral aminoalkanolic derivatives by the connection of the same building block (1), with two amino alcohols. The xanthone (1) was bonded to (R)-(-)-1-amino-2-propanol and (S)-(+)-1-amino-2-propanol to give the chiral derivatives 2 and 3 (Fig. 1). These reactions were carried out with the coupling reagent O-(benzotriazol-1-yl)-N-N-N’-N’-tetramethyluronium tetrafluoroborate (TBTU).

O O O O 1eq TBTU, dry THF CH (2) R: NH 3

OH R OH

OH MeO O MeO O H2N CH (3) R: NH 3 (1) (2) or (3) Fig. 1: Synthesis of chiral 1-amino-2-propanol xanthone derivatives (2) and (3). OH

Considering compound (1), as building block, the expected compounds for a similar reaction are 2 and 3 (Fig. 1) and their structure will be established by spectroscopic methods (1H NMR, 13C NMR, IV and mass spectrometry) and also will be evaluated for enantiomeric purity. All compounds will be evaluated for their neurological activity on nerve conduction properties using the sciatic nerve model [3]. We hope that this methodology lead to the discovery of new chiral compounds with potential antiepileptic activity.

References: [1] Pinto, M.M.M., Sousa, M.E. and Nascimento, M.S.J. (2005), Xanthone Derivatives: New Insights in Biological Activity, Current Medicinal Chemistry, 12 (21), 2517-2538. [2] Marona, H. (1998), Synthesis and anticonvulsant effects of some aminoalkanolic derivatives of xanthone, Pharmazie, 53, 672-676. [3] Mert et al., (2003), Differential Effects of Lidocaine and Tramadol on Modified Nerve Impulse by 4-Aminopyridine in Rats, Pharmacology, 69, 68-73. Acknowledgments: FCT (I&D, nº226/94), FEDER, POCI for financial support.

168

Chiral Alaninol Xanthone Derivatives: Synthesis, Structural Elucidation and Biological Activities

A. Pinto1, C. Fernandes1,2, M. Pinto1,2, A. Pozzi3, B. Matos3, L. Oliveira3 and P. Correia- de-Sá3

1 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy, University of Porto, Portugal. 3 Unit for Multidisciplinary Investigation in Biomedicine (UMIB), Abel Salazar Biomedical Sciences Institute (ICBAS), University of Porto, Portugal.

Chiral compounds are of great interest and comprise an area in continuous development in Medicinal Chemistry [1]. Considering the group of compounds of the chemical family of xanthone derivatives, there is a large variety of natural and synthetic compounds, described in the literature [2] but only a few examples with a chiral moiety [3]. Some of these compounds, namely chiral aminoalkanolic derivatives, have demonstrated important activities in the central nervous system (CNS), with the available data suggesting potential antiepileptic and antidepressant activities [3]. In this presentation it is shown the synthesis of two chiral aminoalkanolic derivatives by the connection of the 2-carboxy-6-methoxyxanthone (1), as building block, with both enantiomers (S and R) of the amino alcohol alaninol (Fig. 1). These reactions were carried out with the coupling reagent O-(benzotriazol-1-yl)-N-N-N’-N’-tetramethyluronium tetrafluoroborate (TBTU). O O OH N TBTU,THF, triethylamine H (S)-(+)-2-alaninol MeO O O O (2) OH O O MeO O OH N TBTU,THF, triethylamine (1) H MeO O (R)-(-)-2-alaninol

Fig. 1: Synthesis of chiral alaninol xanthone derivatives (2) and(3) (3). Considering compound (1), as building block, the expected compounds for a similar reaction are 2 and 3 (Fig. 1) and their structure will be established by spectroscopic methods (1H NMR, 13C NMR, IV and mass spectrometry) and also will be evaluated for enantiomeric purity. Preliminary pharmacological tests concerning sciatic nerve conduction experiments [4] were developed and suggest that this family of compounds is potentially interesting. These preliminary results will be presented in this meeting by Matos and collaborators. We expect that the synthesis of these new chiral xanthone derivaties will result in the discovery of some “hit compounds” with important activities in the central nervous system. [1] Caner, H., Groner E., Levy L. (2004) Drug Discovery Today, 9 (3), 105-110; [2] Pinto, M.M.M. et al. (2005), Current Medicinal Chemistry, 12, 2517-2538; [3] Jastrebska-Wiesek, M., Librowski, T., Czarnecki, R., Marona, H. and Nowak, G. (2003), Polish Journal of Pharmacology, 55, 461-465; [4] Mert et al., (2003), Pharmacology, 69, 68-73. Acknowledgements: FCT (I&D, nº226/94), FEDER, POCI, U. Porto, and Caixa Geral de Depósitos for financial support.

169

Primary Hyperparathyroidism: the unknown world of sporadic cases and their genetic alterations

Alvelos M. 1,3, Barbosa E.2 and Soares P.3,4

1Faculty of Science, University of Porto, Portugal. 2 Biomedical Institute Abel Salazar, University of Porto, Portugal. 3Institute of Molecular Pathology and Immunology of University of Porto, Portugal. 4 Department of Pathology, Medical Faculty, University of Porto, Portugal.

Hypercalcemia is a condition characterized by elevated calcium levels in the blood. It is a common under diagnosed metabolic abnormality and a health problem that affects, in Portugal, about 10.000 individuals. Approximately 90% of the cases are due to primary hyperparathyroidism (PHPT). The majority of PHPT cases are sporadic and related with parathyroid adenoma (80-85%), hyperplasia (15-20%) or carcinoma (1%). About 10% of the PHPT cases are hereditary forms that include MEN 1 gene mutations (Multiple Endocrine Neoplasia type 1) and RET gene mutations (Multiple Endocrine Neoplasia type 2). Recently, it was described a germ-line mutation in the CDKN1B gene in a MEN1 mutation-negative patient with parathyroid and pituitary tumors. The molecular mechanisms underlying the pathogenesis of sporadic PHPT are incompletely understood although alterations in MEN1, HRPT2 and CDKN1B genes have already been reported. In the present work, we aim to perform the genetic characterization of apparently sporadic PHPT cases. [1,3] Constitutional DNA samples were obtained from thirty apparently sporadic PHPT patients. Germ-line mutations in RET, MEN1 and CDKN1B genes were searched by PCR/SSCP and direct DNA sequencing. Detection of somatic alterations involving Cyclin D1 expression was performed by immunohistochemistry.[2] Only polymorphic alterations were detected in RET and CDKN1B genes. These results confirm that none of the cases in our series corresponds to hereditary forms of MEN2 or MEN4. In one case a MEN1 germ-line mutation was detected, showing that this patient harbors, in fact, a familial form of MEN1 instead a sporadic form of PHPT. The Cyclin D1 immunohistochemistry study revealed that four of the 29 (14%) cases analyzed had nuclear staining in 20-35% of the tumor cells, raising the possibility of Cyclin D1 mutation in sporadic PHPT. In conclusion, these results showed that all, except one case, of our series probably corresponds to true sporadic forms of PHPT. Moreover, it was observed an overexpression of Cyclin D1 in 14% of the studied cases. Further studies are in course in order to fully characterize the molecular alterations underlying PHPT. References: [1] Miedlich, S., Krohn, K. and Paschke, R. (2003), Update on genetic and clinical aspects of primary hyperparathyroidism, Clinical Endocrinology, 59, 539-554. [2] Hsi, E.D., Zukerberg, L.R., Yang, W., and Arnold, A. (1996), Cyclin D1/PRAD1 expression in parathyroid adenomas: an immunohistochemical study, Journal of Clinical Endocrinology and Metabolism, 81(5), 1736-1739. [3] Pellegata, N. S.,Martinez, L.Q., Siggelkow, H.,Samson, E., Blink, K., Höfler, H., Fend, F., Graw, J. and Atkinson, M. J. (2006),Germ-line mutations in p27 kip1 cause multiple endocrine neoplasia syndrome in rats and humans, PNAS, 103(42), 15558-15563.

170

Flow cytometric detection of P-gp mediated drug efflux: a method for the evaluation of the activity of P-gp inhibitors

A. Palmeira 1,2,3, R. Lima3,4, H. Seca3, M.H. Vasconcelos3,4, E. Sousa1,2, M. Pinto1,2,

1 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal. 2 Research Center of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), University of Porto, Portugal. 3 Cancer Biology Group, IPATIMUP - Institute of Molecular Pathology and Immunology of the University of Porto, University of Porto, Portugal. 4 Department of Microbiology, Faculty of Pharmacy, University of Porto, Portugal.

Resistance to chemotherapy is a major problem in the treatment of many cancer patients. One of the most studied mechanisms of drug resistance is that mediated by P-glycoprotein (P-gp). P-gp is a multidrug membrane transporter coded by the MDR1 gene and is responsible for the efflux of some drugs from cancer cells. P-gp inhibitors, capable of reversing the efflux of anticancer drugs, are already available for clinical use but show limited success. The synthesis and screening of new small molecules with potent activity as P-gp inhibitors may be of clinical benefit. The aim of the present work was to establish the already described flow cytometric method to detect drug efflux from cells [1, 2] in our laboratory. Two chronic myeloid leukemia cell lines in blastic phase were used. One was a cell line sensitive to drugs (K562 cell line) and another was a cell line resistant to drugs due to overexpression of P-gp (K562Dox cell line), which was obtained by other scientists by long-term exposure of K562 cells to a drug. The levels of P-gp in both cell lines were analysed by Western blot. The accumulation and efflux of Rhodamine 123 (Rh123, a known P-gp fluorescent substrate) were determined by flow cytometry along a time course in the K562Dox cells. Verapamil (a known P-gp inhibitor) was used to confirm our capacity to detect drug efflux in this cell line. We confirmed that K562Dox cells express P-gp, whereas K562 cells do not. We were capable of detecting drug efflux in the K562Dox cells by flow cytometry. Flow cytometric analysis showed that Rho123 accumulation and efflux was blocked by Verapamil. However, the effect of Verapamil was neither total nor long-lasting. Future work will consist of designing small molecules by computational chemistry, as potential inhibitors of P-gp. This will be followed by the synthesis and finally the screening of those new molecules, by using this drug efflux method. References: [1] Ludescher, C. et al. (1991), Blood, 78 (5), 1381-1390. [2] Huet, S., et al. (1998), Cytometry (Communications in Clinical Cytometry), 34, 248-256. Acknowledgements: The authors would like to acknowledge Prof. Dr. Jean-Pierre Marie for the K562Dox cell line, Prof. Dr. José Eduardo Guimarães for helpful discussions and FCT (I&D, nº226/94), FEDER, POCI, U. Porto, and Caixa Geral de Depósitos for financial support.

171

Effects of etoposide, doxorubicin and cytarabine in Burkitt Lymphoma cell lines

R. Lima1,2, H. Seca1, M. I. Castro 1,2, S. Brás1, P.Soares1,3, M.S. Nascimento2,4, M. H. Vasconcelos1,2

1 Cancer Biology Group, IPATIMUP- Institute of Molecular Pathology and Immunology of the University of Porto, University of Porto, Portugal. 2 Department of Microbiology, Faculty of Pharmacy, University of Porto, Portugal. 3 Faculty of Medicine, University of Porto, Portugal. 4 Research Center of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), University of Porto, Portugal.

Epstein-Barr Virus (EBV) infects more than 90% of the world population and it persists as a lifelong infection mainly without symptoms. EBV is usually kept in latent form, possibly leading to cellular transformation. In fact, EBV infection has already been associated with the development of some neoplasias, namely Burkitt Lymphoma (BL). Traditionally, treatment of this type of cancer is achieved using cytotoxic drugs. The aim of this work was to verify the effect of some cytotoxic drugs such as etoposide, doxorubicin and cytarabine in BL (EBV positive and EBV negative) cells in culture. To address this question two isogenic BL cell lines were used: the AKATA EBV negative cell line and its parental AKATA EBV positive cell line [1,2]. Both cell lines were treated over 48h with etoposide, doxorubicin and cytarabine. Cellular viability was analysed with the Trypan Blue exclusion assay. Apoptosis levels were assessed with the TUNEL assay and the expression of apoptotic proteins was verified by Western Blot. The EBV positive cells were more sensitive to etoposide and doxorubicin than the EBV negative cells, but their response to cytarabine was similar. Basal programmed cell death levels were higher in the EBV positive than in the EBV negative cells. In agreement with this, EBV positive cells had higher levels of cleaved PARP than the EBV negative cells. Both cell lines had an increase in programmed cell death following treatments with all drugs. In the EBV positive cells, Bcl-2 expression was lower than in the EBV negative cells and decreased with all treatments. We concluded that EBV positive cells had higher basal programmed cell death levels than the EBV negative cells and that they were more sensitive to doxorubicin and etoposide. However, they presented similar response to cytarabine. We are currently investigating if EBV is responsible for the differences observed in the response of the two cell lines.

References: [1] Takada and Ono (1989), J Virol 63,445-449. [2] Shimizu et al. (1994), J Virol 68,6069-6073. Acknowledgments: The authors would like to thank Professor Kenzo Takada for the AKATA cell lines (EBV negative and EBV positive). They would also like to acknowledge Liliana Santos for technical assistance and the Universidade do Porto and Caixa Geral de Depósitos for financial support. R. Lima is recipient of a PhD grant (SFRH/BD/21759/2005) from FCT.

172

Automatic FIA method for determination of iron in environmental samples

E. P. Freitas, F. A. Dias, H. M. Oliveira, M. A. Segundo, S. Reis and J. L. F. C. Lima

REQUIMTE, Serviço de Química-Física, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha, 164, 4099-030, Porto, Portugal Email: [email protected]; [email protected]

Iron is a ubiquitous element in the environment, especially abundant in rocks and soil. In its biogeochemical cycle [1], iron is absorbed by plants as it is an essential element for its growth and survival. In animals, iron is acquired through their diet. It is essential for their metabolism, sustaining the effectiveness of hemoglobin. When these organisms die, the microbial decomposers will break down essential nutrients from the body, including iron, back to the soil. The presence of iron in different environmental matrices and its important role upon life demands its quantification, namely in studies concerning its bioavailability, toxicity and mobility. In this context, the automation of the determination of iron is essential. Flow Injection Analysis (FIA) [2] is an automation technique based on three principles: (1) reproducible sample injection in a flowing carrier stream, (2) controlled dispersion of the sample zone and (3) reproducible timing of its movement from the injector point to the detection system. These principles allowed determination without need of physical or chemical equilibrium. The aim of this work is to provide a FIA system for the spectrophotometric determination of iron in environmental samples (water, soil, etc). The development of the flow method will comprise several steps, including the selection of the chromogenic reagent, the optimization of reaction conditions (reagent concentration, time), and the optimization of the manifold configuration (sample volume, flow rate, reaction coil length). Finally, the accuracy and the precision of the novel methodology will be assessed through application to real samples.

References: [1] Shaked, Y., Erel, Y., and Sukenik, A. (2004), The biogeochemical cycle of iron and associated elements in Lake Kinneret, Geochimica et Cosmochimica Acta, 68 (7), 1439-1451. [2] Ruzicka, J., and Hansen, E.H. (1975), Flow injection analyses. 1. New concept of fast continuous flow analysis, Analytica Chimica Acta, 78 (1), 145-157.

173

Characterization of exotic wood species for finishing with water- borne varnishes

D. Martins 1, V. Lopes3, F. Oliveira3, A. Mendes1, L. Carvalho1, 2

1 LEPAE - Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal. 2 DEMad - Department of Wood Engineering, Polytechnic Institute of Viseu, Viseu, Portugal. 3 CIN – Corporação Industrial do Norte, S.A., Maia, Portugal.

The finishing is the last operation in the manufacture of a wooden object, which consists of an application of a decorative and protective coating, on the surface of the material. The primary function of any wood finish (paint, varnish or stain, for example) is to protect the wood surface, help maintain a certain appearance and provide a cleanable surface [1]. The quality of a finished surface does not depend only on the technical characteristics of the applied product, but also from the application conditions and the state of wood surface. As an organic material derived from trees, wood is a complex heterogeneous polymer composed of cellulose, lignin, hemicelluloses and minor amounts of extractives materials contained in the cellular structure [2]. Wood characteristics are determined by many factors, such as species and growing conditions. More precisely, wood surface is influenced by several intrinsic factors of the material, mainly morphology of polymers, density, texture, roughness, extractives, moisture content and also by process conditions. The analysis of surface quality is crucial for the successful control of quality of the finishing, mostly in case of water-borne products that are increasingly being used, due to environmental regulations. Nowadays, exotic wood species are very appreciated for outdoors and indoors applications in Portugal, but the ability of water-borne varnishes for finishing these exotic species are less known. Among the species more used in Portugal, 13 were chosen of the most important and commercialized species: Afzelia, Oak, Beech, Jatoba, Mahogany, Iroko, Agba, Chesnut, Ipe, Sucupira, Tatajuba, Wengé and Mutényé. So, the aim of this work is to study several wood surface characteristics that could affect the finishing of wood with water-borne varnishes. To characterize wood surface the following parameters were evaluated: surface pH, colour changes, roughness with and without sanding, wettability and water-soluble extractives. These kind of exotic wood species have different properties related to its surface and characteristics, which will influence the product adhesion and final performance of the finishing (durability and mechanical resistance). References: [1] Williams, R.S. (1999), Finishing of Wood, in Wood Handbook, Forest Products Laboratory, Madison, pp. 15-1 – 15-38. [2] Coelho, C. L., Carvalho, L. H., Martins, J., Costa, C., Masson, D., Méausoone, P. J. (2007), Method for evaluating the influence of wood machining conditions on the objective characterization and subjective perception of a finished surface, Wood Sci Technol, published Online First ™. 2007.

174

Microbiological study of the interaction of Outer Membrane Proteins with antibiotics

M. Garrido1, P. Gameiro1,2, P.J. Eaton2 and M. Feio1,2

1 Department of Chemistry, Faculty of Science, University of Porto, Portugal. 2 REQUIMTE

Pathogen resistance to antibiotics due to their extensive use is posing a major problem to public health. Research into better and improved drugs is therefore a primary concern in this filed. This work is part of a wider project that has the final goal of understanding, at the molecular level, the uptake of antibiotics through porins – channel proteins – that have a crucial role not only in the transport of certain antibiotic families but also in the development of resistance. Specifically, it is intended to obtain detailed information about the molecular interactions of antibiotics belonging to the fluoroquinolone family and OmpF. This membrane protein is known to be associated with the transport of these drugs and it is important to understand the interdependence between the transport mechanism and the drug’s efficiency1,3. The microbiological study of the effect of fluoroquinolones upon a collection of E. coli strains2 with mutations in their OmpF channels is being carried out. The microplate minimum inhibitory concentrations (MIC) method is being used to screen the effect of moxifloxacin in order to elucidate the mechanism of cellular transport. Binary and ternary complexes of copper(II) and 1,10-phenantroline with moxifloxacin will also be tested for the improved efficacy attributed to metalloantibiotics1. Atomic Force Microscopy (AFM) will also be used to complement the study of the action for the different drugs in the different E. coli strains allowing the observation of their effect at a morphologic level.

References: [1] Gameiro, P., Rodrigues, C., Baptista, T., Sousa, I., Castro, B. (2007), Solution studies on binary and ternary complexes of copper(II) with some fluoroquinolones and 1,10-phenanthroline: antimicrobial activity of ternary metalloantibiotics, International Journal of Pharmaceutics, 334, 129-136. [2] Prilipov, A., Phale, P., Van Gelder, P., Rosenbusch, J., Koebnik, R. ( 1998), Coupling site- directed mutagenesis with high-level expression: large scale production of mutant porins from E. coli, FEMS Microbiology Letters, 163, 65-72. [3] Neves, P., Berkane, E., Gameiro, P., Winterhalter, M., Castro, B. (2005), Interaction between quinolones antibiotics and bacterial outer membrane porin OmpF, Biophysical Chemistry, 113, 123-128.

175

Interactions of sulindac and its metabolites with phospholipid membranes: an explanation for the peroxidation protective effect of the bioactive metabolite

F. Santos 1, L. Teixeira1, M. Lúcio1, J. L. F. C1. Lima and S. Reis1

1 Serviço de Química-Física, Faculdade de Farmácia Universidade do Porto, Portugal.

Non-steroidal anti-inflammatory drugs (NSAIDs) are the most important therapeutic agents used in the treatment of inflammatory processes. Although the most prominent action of NSAIDs is due to their inhibitory activity against cycloxygenase (COX) enzymes that catalyse the formation of prostaglandins there are also other important non- prostaglandin-mediated effects. These include the NSAIDs antioxidant effect against lipid peroxidation induced by reactive species which are implicated in several pathophysiological processes such as inflammation, cell injury, cancer and death. Since both inflammatory and lipid peroxidation processes are cell-surface phenomena, the possible effects of NSAIDs on model membrane systems were investigated. Sulindac is a sulfoxide prodrug of the therapeutic class of NSAIDs. Following oral administration, it is reduced by the colonic flora to the pharmacologically active sulfide form, which in turn is oxidized to the pharmacologically inactive sulfoxide and sulfone metabolites. Different experiments performed in liposomes and aqueous solution were compared and used to evaluate the protective effect of sulindac and metabolites in lipid peroxidation induced by the peroxyl radical (ROO•) derived from 2,2’-azobis(2- amidinopropane) dihydrochloride (AAPH) and using fluorescence probes with distinct lipophilic properties. Lipid peroxidation using the hydrophilic probe fluorescein was evaluated in lipid and aqueous media. Lipid systems labelled with the fluorescent probe diphenylhexatriene propionic acid (DPH-PA) were used to assess the effects of the drugs on membrane peroxidation simultaneously by fluorescence intensity decay and changes in membrane fluidity by steady-state anisotropy measurements. The location of sulindac and its metabolites within lipid membrane models was determined by fluorescence quenching using the probe (DPH-PA) inserted across the lipid bilayer. In addition, zeta-potential measurements were made to evaluate changes in membrane surface resulting from its interaction with sulindac and metabolites. Steady-state anisotropy measurements were also made to determine possible membrane fluidity changes induced by the drugs assayed. The use of different probes and liposomes as membrane mimetic systems allowed to conclude that membrane lipoperoxidation, is not only related to the scavenging characteristics of the antioxidants, but also to their ability to interact with lipid bilayers. Results indicate that the antioxidant efficiency is linked to the proximity of the antioxidant to the oxy-radical. The active NSAID studied (sulindac sulfide) can penetrate into the lipid bilayer being accessible to protect membrane against oxy-radicals. In contrast the inactive forms studied (sulindac and sulindac sulfone) do not present significant membrane effects and are more able to scavenge radicals in the aqueous media.

176

Evaluation of primer coatings on steel subtracts by a low-cost, fast electrochemical technique

V. Gonçalves 1, 2, A. Mendes 1, 2, J. Machado 3, F. Oliveira 3, J. Nogueira 2, 3, H. Aguilar Ribeiro 1

1 LEPAE – Laboratory for Process, Environmental and Energy Engineering, Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal. 2 Rede de Competência em Polímeros, Faculty of Engineering, University of Porto, Portugal. 3 CIN – Corporação Industrial do Norte, Maia, Portugal.

Companies producing anticorrosive coatings for steel structures often need to test newly formulated products, to determine their actual behavior before large-scale production. In the coating industry, the most widely spread technique for assessing anticorrosive properties of a coating is still salt fog spray test. However, they give very subjective information and are very time and cost demanding. Modern electrochemical techniques, such as the electrochemical impedance spectroscopy (EIS), have made testing of the corrosion degradation behavior of painted metal systems a relatively simple matter, but the time to obtain an indication of the paint quality was still too long (1 week to 1 month) [1]. In the present work we propose the use of an electrochemical test, known as “AC/DC/AC method” [1], to estimate the barrier properties of three different coatings, containing zinc particles as a corrosion inhibitor agent, applied to steel substrates (Fig. 1). As any other protocol designed to assess coating quality, this one includes a mechanism to stress the coating and induce its failure; this is achieved by combining impedance results (AC) and cathodic polarizations (the sample is cathodically polarized to generate an alkaline environment and stimulate delamination at the metal/coating interface). In the present study, the cathodic polarization (DC) was carried out for 2 minutes at a constant voltage of -2 V, followed by a relaxation step to reach a new equilibrium stage at the normal corrosion potential. The impedance of the stressed system was again evaluated. This sequence stress/impedance was repeated six times, for about 2 hours, to assess anticorrosive coatings performance, while similar conclusions were obtained after more than 800 hours by salt fog spray tests. The results of this study yield a performance ranking of the three different coatings, and show the ability of the AC/DC/AC technique to evaluate properly and much faster the anticorrosive properties of paint coatings.

100,000

10,000 Ω 1,000 100

10 Impedance, IZI / / IZI Impedance,

1 1101001,00010,000100,000

Frequency, f / Hz Fig. 1 – Left: Picture of the experimental set-up; Right:Level I BodeLevel II plot (impedanceLevel III modulus vs. frequency) for different coatings applied on metal substrates after exposure to electrolyte. References: [1] García, S.J. and Suay, J. (2006), Application of electrochemical techniques to study the effect on the anticorrosive properties of the addition of ytterbium and erbium triflates as catalysts on a powder epoxy network, Progress in Organic Coatings, 57, 273-281.

177

The Japanese Influence in Modern Western Architecture ─an introduction─

Miki Itabashi

Faculty of Architecture, University of Porto, Portugal.

The present work, carried out at the Faculty of Architecture as a graduation thesis1, proposes to study the issue of influence in architecture, in particular the Japanese influence in the Modern Western architecture. Through the work of some architects who are considered to be relevant to the history of architecture and modern culture, such as Frank Lloyd Wright, Adolf Loos, Bruno Taut and Walter Gropius, an approach to the Japanese culture is to be made, seeking to identify principles of the traditional Japanese architecture that strengthen and assist the formation of modern idea of Western way of living. The Japanese house is the expression of philosophy, lifestyle and aspirations of the Japanese society. Due to the fact that Japan has lived a long period of isolation from the West, its culture is deeply rooted in itself. The traditional Japanese housing is an example that illustrates, through its simplicity, an adaptable and flexible way of inhabiting, with a close relationship with Man and Nature. The analysis of some Frank Lloyd Wright and Adolf Loos’s houses serve as a motive to establish common points between their architecture and the Japanese one. The study of Bruno Taut and Walter Gropius’s writings will help to the understanding of the traditional Japanese concepts under a modern perspective and its applicability in the contemporary housing. This confrontation of two different cultures will serve to present some basis on Japanese housing and Japanese ways of living and, at the same time, to understand what western architects have learned from it, or have seen, in that completely different culture, as a practical example of concepts that they had already been developing. Besides the comparison between the West and the East, whose purpose is to establish connections between the two cultures and to understand how the knowledge of other architectures can enrich us, both by the valuing of our own culture and by adding new means to solve contemporary architecture problems this study also intends to show how important influence is in projecting, as a tool that, in a conscious or unconscious way, helps us in the search and in the act of creating. This study may also be included in the perspective of reconsidering the modern architecture’s historiography that, as it is known today, does not lay on the assumption of “blank slate” concept, isolated from the past and tradition, as its supporters claimed, when opposing to eclecticism. This work, within its limitations, may be a contribute to the analysis and understanding of history, as well as a disclosure of not so known aspects of this modern western architecture remarkable names.

[1]ITABASHI, Miki, A Influência Japonesa na Arquitectura Moderna Ocidental. Uma Introdução., Prova Final de Licenciatura em Arquitectura, FAUP, Porto, 2007

178

Purinergic signalling in a model of overactive urinary bladder

Nuno Silva1, Miguel Faria1, Vítor Cavadas2 and Paulo Correia-de-Sá1

1Laboratório de Farmacologia e Neurobiologia, UMIB, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS) – Universidade do Porto (UP). 2 Serviço de Urologia, Hospital Geral de Santo António (HGSA), Porto, Portugal.

The mechanisms for relaxation of the bladder during the filling phase, and for initiation of the micturition reflex, are incompletely understood. Normal bladder contraction in humans is mediated mainly through stimulation of muscarinic cholinergic receptors in the detrusor muscle (for a review, see [1]). Atropine-resistant (noncholinergic, nonadrenergic [NANC]) contractions have been reported in normal human detrusor. The atropine-resistant purinergic (P2X1) component of human bladder contraction may be increased to 40% in elderly people and patients with bladder pathologies (e.g. hypertrophic and neurogenic bladders, interstitial cystitis) [2,3]. The bladder epithelium releases ATP in response to mechanical stimuli (and to chemical irritants), and it is hypothesized that ATP released from the serosal surface of the urothelium during bladder filling stimulates P2X3-containing receptors on suburothelial sensory nerve fibres, thus signaling information about urinary bladder filling. Thus, we aim at investigating the mechanisms involved in the control of ATP release from the urothelium, to prompt for new targets (e.g. mechanosensory pathway) for pharmacological manipulation of bladder overactivity. Experiments were carried out, at 37ºC, in urethane-anaesthetized (25% solution, initial dose: 1.0−1.2 g/kg, maintenance dose: 0.1 g/kg), spontaneously breathing male Wistar rats (300−450 g). The dome of the urinary bladder was catheterized using a three-barrel cannula for continuous measurement of intraluminal pressure. The micturition reflex was evoked by bladder distension induced by intravesical infusion of saline (0.05 ml/min). Respiratory tidal volume, bladder pressure, ECG and pelvic nerve activity were continuously displayed on computer screen via a PowerLab data acquisition system (Chart 5, v.4.2 software; AD Instruments, USA). Urine samples before and after bladder filling were assayed for ATP content using the luciferin-luciferase bioluminescence assay (Enliten ATP kit, Promega, USA). Bladder overactivity was induced by intravesical infusion of acetic acid (0.2-1%, v/v in saline). Bladder filling is normally followed (within a few minutes) by an increase in the electrophysiological activity (firing rate and amplitude) of the pelvic nerve, which occurs synchronously to large amplitude spontaneous contractions of the detrusor – micturition reflex. Acetic acid (0.2-1%, for 15 min) concentration-dependently decreased the time (ranging from 67 to 81% of control) and the pressure threshold (ranging from 58 to 85% of control) for the appearance of the first isovolumetric contraction. The magnitude of isovolumetric contractions tends to increase proportionally to the concentration of acetic acid infused into the bladder. During the micturition reflex, we observed an increase (52±7%, n=5) in urinary ATP, which was significantly (P<0.05) amplified in the presence of acetic acid (1%, 297±25%, n=3). The activity of urinary lactate dehydrogenase (LDH) was virtually unchanged in the presence of acetic acid (1%). Although there is extensive literature to indicate that many different types of purinoceptors are present in the lower urinary tract, the pathophysiological role of these receptors in micturition (or voiding) is still uncertain. Our results showed that ATP released from the urothelium may play a role on bladder overacitivity induced by acetic acid (a model of interstitial cystitis). [1] Andersson & Hedlund (2002) Urology 60:13; [2] Ruggieri (2006) Nature Clin Pract Urology 3:206; [3] Yoshida et al. (2004) Urology 63:17. This work was supported by University of Porto / Caixa Geral de Depósitos.

179

Conduction block in isolated rat sciatic nerve by chiral aminoalkanolic derivatives of xanthones

Bernardo Matos1, Angelo Pozzi1, Ana Pinto2,3, Inês Teixeira2,3, Emidio Fernandes1, David Moreira1, Carla Fernandes2,3 , Laura Oliveira1, Madalena Pinto2,3 & Paulo Correia-de- Sá1

1Laboratory of Pharmacology and Neurobiology, UMIB, Institute for Biomedical Sciences of Abel Salazar (ICBAS), 2 Research Centre of Organic Chemistry, Phytochemistry and Pharmacology of the University of Porto (CEQOFFUP), Faculty of Pharmacy; 3 Department of Organic Chemistry, Faculty of Pharmacy, University of Porto, Portugal.

The chemical family of xanthone derivatives was described to be one group of compounds with a broad spectrum of biological activities [1]. In fact, it has been demonstrated that some chiral aminoalkanolic derivatives of xanthones possess potential antiepileptic and antidepressant activities, however, their mechanism of action at the neuronal cell level is largely unknown [2]. This prompted us to synthetise new chiral alaninol xanthone derivates and to evaluate their pharmacological activity in the nervous system. Since, these coumpounds can potentially act as modulators of Na+ ionic currents, their neurologic activity was evaluated on the nerve conduction properties of rat sciatic nerve. Electrophysiological experiments were carried out on sciatic nerves isolated from Wistar rats (200-250 g). We used a simple extracellular partition recording technique for monitoring chiral alaninol xanthone derivates-induced changes in membrane polarization and evoked potential amplitudes in sciatic nerve preparations [3]. Sciatic nerves were stimulated supramaximally with 0.05 ms duration square-wave pulses delivered at a frequency of 0.2 Hz (Tonic stimulation). Chiral alaninol xanthone derivates were applied during 30 minutes and their influence on nerve compound action potentials (CAP) were recorded. All CAP parameters (amplitude, half width, and depolarization time) were analysed. CAP parameters induced by test drugs were reported as a percentage of the control values (in the presence of Tyrode’s solution). CAP amplitude, half width and depolarization time observed in control conditions were 9.4±4 mV (n=3), 0.69±0.05 ms (n=3), 1.28±0.04 ms (n=3), respectively. Chiral alaninol xanthone derivates (XEVOL and XEA, 0.100-10 μM) increased nerve conduction block, in a concentration-dependent manner. When applied in a concentration of 1 µM, XEVOL and XEA decreased sciatic nerve conduction by 68±11% (n=3) and by 40±21% (n=2), respectively. Pre-treatment with XEVOL did not significantly (P>0.05) change both the depolarization time (19±13%, n=2) and the half width (17±12%, n=2), even when the highest concentration (10 μM) was used. Nerve conduction block may indicate that chiral alaninol xanthone derivates possess membrane stabilizing properties, probably by decreasing Na+ ionic currents. Among the xanthone derivates tested, XEVOL was more potent than XEA. Whether these compounds have potential for treating epileptic seizures, neuropathic pain and/or bipolar disorders requires further investigation. [1] Pinto et al (2005), Curr Med Chem, 12, 2517-2538; [2] Jastrebska-Wiesek et al (2003), Pol J Pharmacol, 55, 461-465; [3] Mert et al (2003), Pharmacology, 69, 68-73. This work was supported by University of Porto / Caixa Geral de Depósitos.

180

Osteoclastic resorption of calcium phosphate based bone substitutes: in vitro studies

C.A. Teixeira1,2, J. Costa-Rodrigues1, P.S. Gomes1, Almeida Palmas, R. and M.H. Fernandes1

1 Laboratório de Farmacologia e Bicompatibilidade Celular, Faculty of Dental Medicine, University of Porto, Portugal. 2 Faculty of Sciences, University of Porto, Portugal.

Bone is a dynamic tissue that undergoes constant remodelling, a process that requires a perfect coordination between bone synthesis (mediated by osteoblasts) and bone resorption (mediated by osteoclasts) [1]. Osteoclasts are specialized multinucleated cells derived from the monocyte haematopoietic lineage. They adhere to bone matrix and promote bone resorption through the secretion of acid and lytic enzymes. Deficiencies in osteoclast number/function can lead to osteopetrosis, a disease characterized by bone deformities caused by abnormal quantities of non-remodeled bone mass. On the other hand, increased number and activity of osteoclasts may cause accelerated bone resorption, wich can lead to osteoporosis (reviewed in [2]). An essential property of bone substitutes is that they are integrated into the natural bone remodelling process. Synthetic bone substitutes have numerous applications in medicine and dental medicine. They promote bone repair and regeneration which makes them useful tools in orthopedy, periodontal and maxillofacial surgery, and implantology. Calcium phosphate based materials are the most commonly used, due to similarities with mineral bone matrix. Materials implanted in bone tissue must have adequate surface properties for normal cell activity of bone remodelling. After implantation there is recruitment of osteoblastic precursors to the material surface, followed by proliferation and differentiation, wich leads to bone formation. This process is essential for osteointegration of the material. However, the material should also allow a normal osteoclastic activity and must be resorbed by cellular mechanisms. The balance between these two processes is essential for long term survival of the implanted material [3]. In this work we will evaluate the proliferation and function of osteoclastic cells isolated from human peripheral blood and cultivated in calcium phosphate based bone substitutes with different surface characteristics. The characterization of osteoclast cultures in the different conditions will be based on different parameters, namely: cell morphology and adhesion, formation of multinucleated cells (osteoclastogenesis), formation of actin rings, presence of vitronectin receptors (αVβ3 integrin), expression of specific osteoclastic genes, tartrate resistant acid phosphatase activity, and formation of resorption lacunae.

References: [1] Schilling A.F., Linhart W., Filke S., Gebauer M., Schinke T., Rueger J.M., Amling M. (2004) Resorbability of bone substitute biomaterials by human osteoclasts. Biomaterials. 25 (18), 3963- 3972. [2] Boyle W.J., Simonet W.S., Lacey D.L. (2003) Osteoclast differentiation and activation. Nature. 423 (6937), 337-342. [3] Shen Z., Crotti T.N., McHugh K.P., Matsuzaki K., Gravallese E.M., Bierbaum B.E., Goldring S.R. (2006) The role played by cell-substrate interactions in the pathogenesis of osteoclast-mediated peri-implant osteolysis. Arthritis Res. Ther. 8 (3), R70.

181

Monitoring particulate matter using magnetic properties of tree leaves in the Porto urban area and Cinfães area

H. Corrêa-Ribeiro1

1 Department of Geology, Faculty of Sciences, University of Porto, Portugal.

Magnetic particles due to emissions by the road traffic and others human activities such as metallurgical, cement industries, power stations burning fossil fuels and fires are the main a cause of cardiorespiratoire diseases. The magnetic susceptibility of tree leaves has been used as a proxy to evaluate the particulate matter quantity and the road traffic contribution. In order to evaluate pollution levels, sampling zones were chosen in the urban area of Porto and compared with the rural area of Cinfães. Sampling was carried out in the first week of June, July and August 2006. Leaves from Nerium oleander, Quercus sp., Tilia sp. And Platanus sp. were sampled on a total of 32 sites in Porto and 48 sites in Cinfães area. Magnetic susceptibility measurements were performed using an Agico Kappabridge (KLY-4S, Geology Department, U.P.) and the values calculated by mass. In Porto area magnetic susceptibility mass (χ) was comprised between -4.47 and 118.22 E-9 m3/kg and only one site reveals diamagnetic values. All the others present high positive susceptibility values which indicate the presence of anthropogenic ferromagnetic particles. In Cinfães area χ is lower and comprised between -8.13 and 13.89 E-9 m3/kg but the most sites presented diamagnetic values with exception for the sites located within Cinfães village. In August a decrease in susceptibility values was observed in Porto, probably due to the reduction of road traffic during holidays. Quite the opposite was observed in Cinfães area, in the same period, an increase of χ, due to the road traffic rise during holidays and also to the fires that occurred in August. Significant positive correlation exist between χ and the contents of Fe and Cu of leaves, showing that the magnetic properties of leaves can be used as a preliminary index for detection of Fe and Cu pollution. These results indicate that a magnetic survey of tree leaves, which is relatively quick and inexpensive, may be used in addition to the municipal air quality monitoring systems to identify high-polluted areas in urban and non-urban environments.

Keywords: environmental magnetism, magnetic susceptibility, chemical analysis.

182

Adenosine regulates its own release from myenteric neurons via A2A receptors coupled to adenylate cyclase / cAMP pathway

J. Duarte, R. Dias, M. Duarte-Araújo, T. Magalhães-Cardoso & P. Correia-de-Sá

Laboratório de Farmacologia e Neurobiologia, Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar - Universidade do Porto (ICBAS-UP), Portugal.

Adenosine is a ubiquitous component of cells, which acts as a homeostatic regulator in the nervous system [1]. In the gastrointestinal tract, besides the inhibitory effect of adenosine mediated by neuronal A1 receptors [2], endogenous adenosine plays a predominant facilitatory action on ACh release from myenteric neurons of the rat ileum, via the activation of prejunctional facilitatory A2A receptors [3]. Myenteric neurons are the main source of adenosine released per se in response to electrical stimulation [4]; parallel formation of adenosine from the hydrolysis of released ATP (via ectonucleotidases pathway) may also occur during prolonged periods of stimulation [3]. It remains, however, to be elucidated whether adenosine plays a role regulating its own extracellular levels. The experiments were performed at 37ºC on longitudinal muscle-myenteric plexus (LM-MP) of rat ileum, superfused with gassed (95% O2 + 5% CO2) Tyrode´s solution. LM-MP preparations were stimulated (10 Hz, 3000 pulses, 1 ms, 100 V) in the presence of test drugs. Samples were collected before and after stimulus application and retained for nucleoside analysis by HPLC. Electrical stimulation of the LM-MP increased (74±6%, n=6) the outflow of adenosine (ADO) above control levels. Blockade of action potentials generation with tetrodotoxin (1 µM) or omission of Ca2+ (plus EGTA, 1 mM) in the buffer essentially abolished the release of ADO. Blockade of smooth muscle contractions by inhibiting Ca2+ influx through L-type channels, with nifedipine (1 µM), was devoid of effect on ADO outflow. Application of the adenosine kinase inhibitor, 5’-iodotubericidin (ITU, 10 µM), led to an increase in stimulation-induced ADO accumulation by 51±6%, (n=8). The nucleoside transport inhibitor, dipyridamole (DIPY, 0.5 µM), was more effective in inhibiting (-59±9%, n=4) ADO accumulation in the stimulated LM-MP as compared with the ecto-5’-nucleotidase inhibitor, concanavalin A (Con A, 0.1 mg/ml, -27±1%, n=5). Activation of A2A receptors with CGS 21680C (3 nM) increased stimulation-induced ADO outflow by 129±10% (n=5). The facilitatory effect of CGS 21680C (3 nM) was prevented by the selective A2A antagonist, ZM 241385 (50 nM) and by DIPY (0.5 µM), while Con A (0.1 mg/ml) was unable to change the effect of CGS 21680C (3 nM). The adenylate cyclase activator, forskolin (FSK, 3 µM), mimicked (158±14%, n=4) the facilitatory effect of CGS 21680C (3 nM) on the release of the nucleoside; the effect of FSK (3 µM) was also attenuated by DIPY (0.5 µM). These results indicate that although ADO may be formed by the extracellular catabolism of released adenine nucleotides via the ectonucleotidase pathway, this might not represent the major source of endogenous ADO in the rat myenteric plexus. Normally, Ca2+-dependence release of neuromediators indicates a vesicular storage and exocytotic release. Since there is no evidence for vesicular storage and release of ADO, the intracellular Ca2+ requirement for ADO release may reflect actions on intracellular processes that modify metabolic production of the nucleoside or promote its efflux through equilibrative transporters. Data showed that ADO mediates a positive feedback mechanism regulating its own transport from myenteric motoneurons into the extracellular space via the activation of A2A receptors coupled to adenylate cyclase / cyclic AMP pathway. [1] Cunha (2001) Neurochem. Int., 38, 107-125; [2] De Man et al. (2003) Br. J. Pharmacol. 139, 172-184; [3] Duarte-Araújo et al. (2004) Br. J.Pharmacol. 141, 925-934; [4] Correia-de-Sá et al. (2006) Auton Neurosci. 126-127, 211-224. This work was supported by FCT (FEDER funding, POCTI/45549/FCB/2002) and PTDC/74462/CVT/2006 projects).

183

Education and Poverty: an empowerment approach

A. Oliveira1

1 Department of Educational Sciences, Faculty of Psychology and Educational Sciences, University of Porto, Portugal.

This presentation settles on the research in course for writing the thesis for the European Master of Development Studies in Social and Educational Sciences, specialization in “European Perspectives on Social Inclusion”. The theme relates education with poverty through an empowerment approach, taking for study a European Union (EU) food help programme and Adults Education and Training courses promoted by Care Institutions. It is being used for the research a comparative method. There are taken two Care Institutions that distribute food from the EU food help programme and that are situated on Porto’s District in Portugal. One institution promotes in relation to the food help programme, Adult Education and Training courses and the other doesn’t. For methodological techniques it will be used interviews with the directors of the food help programme in the care institutions and a questionnaire applied to the people that receive those benefits. Although poverty is a multidimension concept [1], the option for taking education as the comparative focus lies on the believe that it has the potential for promote the necessary empowerment for the individuals to improve their live conditions. Education is only a helping point given that there are several other dimensions that influence our live, and in this case, poverty condition. We can point, for instance, family or self ownership, residence territory, job conditions, housing, health, education, power and participation. There can be established relations between all these dimensions in what concerns to poverty condition, nevertheless, education was chosen on the basis of its relation with power and participation and with the ability to be aware of rights and duties and so, the dimension which provides more autonomy to the individuals. The results expected to be found, as it is still an on-going research, point to better outcomes in what respects to the goal of fighting against poverty expected from the food help programme in the case in which it was associated with Adult Education and Training courses. These results can be examined through the self-declared improvement in live conditions of the beneficiaries of the food help programme emerged from the fact that they assisted an Adult Education and Training course.

References: [1] Rodrigues, F., Constantin, T., Van Den Hoven, R. and Nunes, M.H. (2005), European Perspectives on Poverty and Poor People / Pobreza e Perspectivas Europeias, Peter Lang, Frankfurt am Main.

184

Synthesis of vesiculated polyester particles and its incorporation in paints

A. Dias 1, A. Mendes 2, F. Oliveira3, J. Moniz4 and F.D. Magalhães2

1 Rede de Competência em Polímeros, Porto, Portugal. 2 Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal. 3 CIN -Corporação Industrial do Norte, S.A., Maia, Portugal. 4 Resiquímica -Resinas Químicas, S.A., Lisboa, Portugal.

Titanium dioxide is a white pigment which confers to paints high whitening index and opacity due to its capability of scattering the light and low absorption power. However, titanium dioxide is currently one of the most expensive raw materials used in a paint formulation, so it is very important for the paint industry to reduce its quantity in paints. Vesiculated particles (VP) consist of an aqueous emulsion of reticulated polyester particles that incorporate numerous water-filled spaces [1]. When a film made with these particles is dried, the water contained in the core evaporates creating air voids (Figure 1), which scatter the light, due to the difference in refraction index between the polymer and the air domains. This film will therefore appear white to the eye. Then, VP are suitable for the beneficial replacement of titanium dioxide present in paint formulation. The process for the production of VP begins with the formation of a first emulsion in which an aqueous phase is emulsified in a mixture of the unsaturated polyester, styrene and a base. The resulting “water-in-oil” emulsion is then itself emulsified and stabilized into an aqueous phase, at high shear. Then, a free radical polymerization is initiated at room temperature, which leads to the reticulation of polyester with styrene and the subsequent production of solid vesiculated particles. The VPs obtained by this double emulsion have a solid content of 13% and a mean particle diameter of 5 μm.

Fig.1. Cryo-SEM image of air voids in vesiculated particles.

The stability of the first emulsion was studied by adding different kinds of bases at different concentrations. It was possible to emulsify 150% of water into the polyester phase by adding triethanolamine in a 0.5 mmol/g concentration. The base must be added to the oil phase to react with the carboxylic group of the polyester to form at the waterpolymer interface polyester salts, which act as true emulsifying agents [2]. This emulsion can be further optimized in the presence of a surfactant. The stability of the second emulsion is also being studied, in terms of the optimal stabilizers concentration. In addition, the conditions for the final reticulation process are also being optimized. References: [1] Engelbrecht, J. et al. (2006), Vesiculated polymer particles, US patent 2006/0111474. [2] Horie, K., Mita.I. and Kambe H. (1967), Copolimerization of unsaturated polyester with styrene in inverted emulsion, Journal of applied polymer science, 11, pp. 57-71.

185

Characterization of Urea-Formaldehyde Resins

F. Silva 1, J. Ferra 2, L. Carvalho 2,3, J.P. Liberal 4 and F. D. Magalhães 2

1 RCP – Rede de Competência em Polímeros, Porto, Portugal 2 Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal 3 DEMad – Department of Wood Engineering, Polytechnic Institute of Viseu, Viseu, Portugal 4 Euroresinas – Indústrias Químicas, S.A., Sines, Portugal

Urea-Formaldehyde (UF) resins are the most widely adhesives used in industry as binders for the production of bonded wood products, particularly particleboards, because of their versatility and low cost, despite of their vulnerability to moisture and water. This type of resins is based in the reaction between the two principal monomers, urea and formaldehyde, originating mixtures of several condensation products like simple methylolureas (mono-, di- and tri-) or insoluble high molecular weight compounds. Nowadays, the major problem to be solved consists in the formaldehyde emission after the curing process, during the lifetime of the material, namely in domestic furniture uses. The emission has to obey certain limits and all efforts and research are directed towards reducing Formaldehyde/Urea (F/U) ratio, maintaining the performance properties of a conventional UF resin. [1] The specific resins being studied in this work are produced by Euroresinas – Indústrias Química, S.A. Since the principal objective of this work is to implement methods of morphological and physical-chemical characterization of the resins, we use two chromatographic methods, HPLC (High Performance Liquid Chromatography) and SEC (Size Exclusion Cromatography), calorimetric methods, DSC (Differential Scanning Calorimetry), and the measure of Particle Size Distribution (PSD), Coulter-Counter. With SEC, we can determine the molecular weight distribution of one resin, separating the molecules according to their hydrodynamic volume which can be proportional to the molecular weight. HPLC allow us to quantify urea and methylolureas (low molecular weight reaction products) present in the resin. DSC studies directly the cure process of the resin, determining the heat flow and the cure temperature, parameters that give an idea of the resin reactivity. Finally, Coulter-Counter, give us the PSD of the insoluble fractions of the resins. The compilation of all these methods also permits the study of the phenomena involved in the resins ageing process. Table 1 shows the parameters range obtained for each one of the above referred techniques.

Table 1. Parameters Range for the different techniques used in the resins characterization. Techniques SEC HPLC DSC PSD

Molecular Weight % Urea TCURE (ºC) (-ΔH) (J/g) Size (mm) Parameters Range 100 - 10000 40 - 60 80 - 90 50 - 80 0.1 - 100

The final UF resins performance is evaluated by producing wood-based panels in a laboratory hot-press. These will be then tested to evaluate several properties: density, moisture content, internal bond, thickness swelling and formaldehyde content. At this moment, all the methods are well implemented, making possible to distinguish different types of resins. To improve the resin formulation, a Design of Experiment methodology will be initiated. References: [1] Dunky, M. (1997), Urea-Formaldehyde (UF) adhesive Resins for wood, Int. J. Adhesion & Adhesives, 18, 95-107.

186

Characterisation of the Planctomycetes Pirellula sp. OJF20 and Pirellula sp. OJF27 isolated from the surface of macroalgae

F. Viana1, J. Torres1, R. Abreu1, J. Bondoso1,2 and O. Lage1,2

1 Departamento de Botânica, Faculdade de Ciências Universidade do Porto, Portugal. 2 CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental

Planctomycetes are a morphologic and phylogenetic distinct group of the domain Bacteria. Members of this group are characterised by budding, cellular compartmentalization and absence of peptidoglycan in the cell wall, features that place Planctomycetes in a very intriguing place in the evolution of microorganisms. Molecular and ecological studies provided evidence about their abundance in a great diversity of habitats underlying their important role in the ecology of the ecosystems [1]. They have been described in probable association with sponges, crustacean, prawns, microalgae and Cyanobacteria. However little is known about this group due to the relatively few species grown in pure culture thus the great importance of isolation and cultivation studies for its better understanding. OJF Planctomycetes culture collection has, presently, about 200 strains, the majority isolated from the surface of different macroalgae [2]. Pirellula sp. OJF20 and Pirellula sp. OJF27 are strains isolated from the surface of Corallina sp. and Condrus crispus, respectively. Corallina was collected in Foz (Porto, Portugal) and C. crispus in Carreço (Viana do Castelo, Portugal), areas with different levels of pollution. Both strains are closely related and placed in a new cluster of Planctomycetes. DNA-based fingerprinting methods (16S rDNA analysis and BOX-PCR), growth in liquid medium and physiological studies (range of temperature, pH, salinity, and nutritional requirements) were performed in order to allow a comparison between the two strains. Despite the 98% 16S rDNA sequence similarity between the two strains that places them in the same species, they present significant metabolic and physiologic differences and are adapted to diverse environmental conditions.

[1] Fuerst, J. A. 1995. The planctomycetes: emerging models for microbial ecology, evolution, and cell biology. Microbiology 141:1493-1506;

[2] Lage OM, Bondoso J. 2007. Marine Planctomycetes-Macroalgae Association from the Atlantic North Coast of Portugal. Presented at Sustainable Neighborhood - from Lisbon to Leipzig through Research (L2L), Leipzig. http://www.fona.de/de/3_akteure/forum_2007/abstracts_poster.php?lang=eng&sort=titel

187

Investigation of the Interactions between Phospholipids and Gold Nanoparticles at Air/water Interface

C. Carreira, F. Silva and C. Pereira

CIQ-L4, Chemistry Department, Faculty of Sciences, University of Porto, R. do Campo Alegre, 687, 4169-007 Porto Portugal.

This study aims to explore the interaction between gold oleylamine modified nanoparticles and phospholipids using monolayers at the air / water interface and how the pH of the subfase influence these interaction. This study can provide insights on the importance of the charge and hydrophilic character of gold nanoparticles on their interaction with model membranes. The combined information from the surface pressure and surface potential isotherms allowed the analysis of the molecular properties of the films. Langmuir films were prepared at the air/water interface using a standard LB trough (KSV mini trough) and the surface potential was measured using the vibrating plate method. The films were prepared spreading 1,3 mg/mL chloroform solution of DPPC on a subphase containing a 30 µL nanoparticles solution. The nanoparticles solution was obtained by dilution of a stock solution with different buffers, and kept at 20 ºC. The monolayer was compressed at the speed of 10 mm/min. The gold oleyl amine functionalized nanoparticles were prepared accordingly to the literature [1]. Figure 1 shows the pressure-area isotherms for the mixed gold nanoparticles and DPPC monolayers, at different pH.

50 pH: 45 8.78 40

-1 6.91 35 2.32 30 4.87 25 12.29 20 15 10

Surface Pressure/ mNm 5 0 30 50 70 90 110 130 150

Mma / Å2 molecule -1

Fig. 1 – Surface pressure isotherm for interaction between gold oleyl amine modified nanoparticles and DPPC at different pH

A decrease of the subphase pH leads to an expansion of the gold nanoparticles/DPPC monolayer. Further results will be presented and discussed.

References: [1] Aslam, M., Fu, L., Su, M., Vijayamohanan, K., Dravid, V.P. (2004) J. Mater. Chem., 14, 1795

188

Development and validation of a new multiresidue method for the determination of 17 polychlorinated dibenzodioxins (dioxins) and polychlorinated dibenzofurans (furans) in environmental matrices by SPME-GC-MS

A.Neves(1), A.D. Guimarães(2), M.F. Alpendurada(1,2)

4450-113 MATOSINHOS – Portugal 1) FFUP – Faculty of Pharmacy, University of Porto, Lab. of Hydrology / Rua Aníbal Cunha, 164- 4050-047 PORTO – Portugal 2) IAREN – Water Institute of the Northern Region / Rua Dr. Eduardo Torres, 229 4450-113 MATOSINHOS – Portugal * Corresponding author: [email protected]

Polychlorinated dibenzodioxins (PCDD) and polychlorinated dibenzofurans (PCDF) are persistent, highly lipophilic and toxic substances. These compounds are widespread in the environment and occur mainly as secondary products of thermal processes involving such as waste incineration, cement kilns firing hazardous waste, production of pulp using elemental chlorine or several metallurgical industry processes. When released into aquatic environments, PCDD/Fs become attached to organic particles which may be adsorbed on suspended matter or even may sink down to the sediments. Eventually they tend to bio accumulate through the food-chain which constitutes the main way of exposure to humans. The harmful health effects of PCDD/Fs on men are evaluated by their action on a cellular receptor, the Aryl Hydrocarbon Receptor (AhR) and include impairment of different systems: immune system, nervous system, hormonal system as well as reproductive functions. PCDD/Fs are also suspected of causing cancer. Solid phase micro extraction coupled to capillary gas chromatography - ion trap mass spectrometry (SPME-GC-MS) is presented as an alternative method to determinate 17 toxic PCDD/Fs. Sensitiveness, quickness, efficiency simplicity and low cost analysis are some advantages of the designed method(1,2). Global analytical method was optimized: extraction step, chromatographic conditions and statistical parameters. Three different of fibres were assayed: 100 µm, 7 µm, polydimethylsiloxane (PDMS) and 75 µm Carboxen- PDMS fibers, and the best result for the majority of the compounds were obtained with 100 µm PDMS. Different extraction times (15, 30, 45, 60 min) and temperatures (70, 90, 100ºC) were studied. The best results were obtained with 60 min of extraction at 90 ºC. Neither pH adjustment nor ionic strength correction were necessaries to obtain good results, which enhances life expectancy of SPME fibre and reduces sample handling Linearity, repeatability, reproducibility, uncertainties, matrix effects, analytical sensitivity and influence of the sample preparation protocol have been studied for method validation in agreement with the international standard ISO/IEC 17025:2005. Bibliographic references: 1-Fabrellas, B.; Sanz, P; Abad, E; Rivera,J.; Larrazábal, D.; Analysis of dioxins and furans in environmental samples by GC-ion-trap MS/MS; Chemosphere, vol.55, 2004; 2-M:F:Alpendurada, Solid-Phase micro-extraction: a promising technique for sample preparation in environmental analysis, J. Chrom.A, 889 (2000) 3-14

189

Adenosine regulates differentiation of human osteoblast cells in culture

A. Barbosa1, M.A. Costa1,2, T. Magalhães-Cardoso1, A. Teixeira1, R. Freitas3, J.M. Neves3 & P. Correia-de-Sá1

1Laboratório de Farmacologia e Neurobiologia e 2Departamento de Química, UMIB, Instituto de Ciências Biomédicas Abel Salazar - Universidade do Porto (ICBAS-UP), and 3Serviço de Ortopedia e Traumatologia, Centro Hospitalar de V.N. Gaia (CHVNG), Portugal.

Bone turnover takes place at discrete sites in the remodeling skeleton that are subject to mechanical loading forces. Extracellular purines are important local modulators of bone cell function. Adenine nucleotides in bone microenvironment are largely determined by ATP release from stressed cells and subsequent metabolism by ecto-enzymes, both of which have been scarcely investigated in the human bone. Breakdown of ATP into adenosine restricts its action to that of a localized signal and shifts purinergic transmission mediated by P2 receptors to long-lasting modulatory signals mediated by P1 adenosine receptors. Surprisingly, there are a few reports of the regulation of cell function by adenosine in bone cells (e.g. Shimegi, 1995, Calcif. Tissue Int., 58:109). This prompted us to investigate the role of stable adenosine analogues in the proliferation and differentiation of non-modified human osteoblast cells in culture. Human bone marrow was collected in sterile conditions from patients that underwent orthopaedic surgery. These proceedings had the approval of the Ethics Committees of CHVNG and ICBAS-UP. First passage bone marrow was cultured in supplemented α-Minimal Essential Medium (α-MEM) for up to 28 days in the absence and in the presence of CPA (30 nM), CGS21680C (10 nM), NECA (0.3 µM) and 2-Cl-IB-MECA (10 nM). Throughout their lifespan, cultures were characterised for morphology, cell viability/proliferation (MTT assay), total protein content (method of Lowry), and alkaline phosphatase (ALP) activity. For the kinetic experiments of ATP catabolism, samples (75 µl) were collected from the bath at different times up to 30 min for reverse-phase HPLC analysis of the variation of substrate disappearance and product formation. Human osteoblast cells in culture, hydrolyse extracellular ATP (30 µM) forming sequentially ADP, AMP and adenosine whose concentrations increased to 1.99±0.18 µM, 0.69±0.04 µM and 17.81±0.64 µM after 30-min incubation, respectively. In control cultures, osteoblast cells proliferated for approximately 2-3 weeks; the maximum values for MTT reduction and total protein content were observed at day 14 (MTT assay, 0.626±0.112, n=9). During the proliferation phase, incubation of osteoblasts with stable adenosine analogues, CPA (30 nM), CGS21680C (10 nM), NECA (300 nM) and 2-Cl-IB.MECA (10 nM), did not significantly change (P>0.05) their ability to reduce MTT. Osteoblast differentiation measured as the activity of ALP on day 14 decreased significantly (P<0.05) from 1.150±0.154 (n=5) in the absence to 0.640±0.141 (n=5), 0.600±0.074 (n=5), 0.590±0.038 (n=5) and 0.480±0,086 (n=5) in the presence of CPA (30 nM), CGS21680C (10 nM), NECA (300 nM) and 2-Cl-IB.MECA (10 nM), respectively. Data indicate that human osteoblasts possess a high ecto-ATPase activity converting sequentially ATP into ADP and AMP, with a significant amount of adenosine being accumulated in the cultures. Stable adenosine analogues consistently affected differentiation of human osteoblasts in culture demonstrated by the reduction of ALP activity, indicating the presence of adenosine receptors in bone cells. Supported by Univ. Porto/CGD and by FCT (FEDER funding, PTDC/SAU- OSM/73576/2006).

190

The Pombais (pigeon houses) in the northeast Transmontano (northeast of Portugal)

Mafalda Matias 1, Teresa Fonseca, PhD Arch 2

1 Faculty of Architecture, University of Porto, Portugal. 2 Faculty of Architecture, University of Porto, Portugal.

Among the vernacular forms of architecture to be found in the Portuguese northeast region of Trás-os-Montes, the “Pombais” (pigeon houses) are a paradigmatic case of a threatened edified patrimony as a unique and synthetic expression of the common sense of utility, present in almost all country settlements. By using the resources available and identifying lands of poor economy, this culture offers us magnificent examples of the wise combination of objectives and assets. Hermetic, sober, austere, ornamental, ornamented also, made of schist or granite, with tile or slate, they all seem to have the only function of providing an address to birds – the pigeons. The “Pombais” belong to the architecture that is non-inhabited by humans yet they evidence an ancestral and only practice of sustainability. They are sophisticated objects of study capable of sustain an investigation in this special matter. Given the scarce information regarding these constructions this investigation develops in different ways - from the bibliographic research to the search of specific information gathered from the real samples of the “Pombais” through the creation of technical records for the different typologies completed in the field with the help of the metric, the GPS, and the camera and military maps, the use of drawing, the interview with qualified and specialised entities and with property owners and local inhabitants, alive testimonies of the popular wisdom, the participation in team exercises of direct action on the object of study. This empirical work allowed a rigorous gathering of the constructive, shaping and architectural characteristics of the “Pombais”, elements that allowed the defence of an architectural type with the potential to be followed, innovated, updated or updatable and, most important of all, published as a model of sustainability. We verify that to be operative, an architecture must adapt to the geographic, social and cultural characteristics of the environment, solving positively to the possible contradictions between the local and the universal. This investigation contributes also to register these constructions in the discipline and scientific areas of architecture as well as in the architectural and cultural patrimony of our country.

191

Pravastatin Quantification using Square-Wave Voltammetry

M. Neves 1, S. Pereira1, H. Nouws2 and C. Delerue-Matos2

1 Departamento de Botânica, Faculdade de Ciências, Universidade do Porto, Portugal 2 REQUIMTE, Instituto Superior de Engenharia, Instituto Politécnico do Porto, Portugal

Nowadays cardiovascular diseases are the main cause of morbidity and mortality in developed countries. Extensive epidemiologic studies reveal that hypercholesterolemia is causally related to an increased risk in atherosclerosis and coronary heart disease. A class of lipid-lowering drugs called statins is the most widely used in hyperlipidemic pharmacological treatment. These compounds reduce levels of circulating atherogenic lipoproteins by competitive inhibition of HMG-CoA redutase, the rate-limiting enzyme in the synthesis of cholesterol. Currently, six therapeutic statins are available on the Portuguese market: atorvastatin, fluvastatin, rosuvastatin, simvastatin, lovastatin and pravastatin. Most statin analysis are performed by liquid-chromatography (LC) coupled with spectrometric detectors. The main disadvantages of LC are the relatively long analysis times, high acquisition and maintenance costs and the use of large volumes of toxic solvents. In the present work an alternative procedure, based on square-wave voltammetry (SWV), for the quantification of pravastatin in pharmaceutical products is proposed. Initial studies showed that pravastatin can be oxidized at a glassy carbon electrode, with a maximum peak current intensity at a potential of +1.3 V vs. AgCl/Ag, in a Britton- Robinson buffer of pH 5 (I = 0.3 mol/L). Several parameters of the SWV method were optimized, resulting in the following conditions: frequency = 90 Hz, step potential = 5.25 mV and step amplitude = 40.05 mV. The method was validated by assessing the linear range between peak current intensity and pravastatin concentration, limits of detection and quantification, repeatability, intermediated precision and accuracy. The developed methodology was applied in the quantification of pravastatin in a pharmaceutical product. It could be a useful alternative to the established methods for this purpose since it provides high sample rates at reduce costs with a low pollution rate.

192

Microbial community distribution and dynamics in a shallow raceway-recirculating mariculture system

A. Matos 1,2, M.-T. Borges 1,2 and P.M.L. Castro 3

1 Department of Zoology-Anthropology, Faculty of Sciences, University of Porto, Praça Gomes Teixeira, 4099-002 Porto, Portugal. 2 CIMAR/CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, University of Porto, Rua dos Bragas 289, 4050-123 Porto, Portugal. 3 Escola Superior de Biotecnologia, Catholic University, Rua Dr. A. Bernardino de Almeida, 4200- 072 Porto, Portugal.

A shallow raceway is an alternative system to traditional tanks for turbot (Scophthalmus maximus) cultivation, promoting higher production. Culture water can be reused (recirculation) if a complex treatment loop, consisting of a mechanical filter, biofilter and ozone disinfection, is used [1, 2]. The aims of this work were to study, during 221 days and from initial colonization, the distribution of the heterotrophic bacterial population in the production system and to evaluate the dynamics of bacteria attached to the biofilter carriers. Viable heterotrophic planktonic and fixed bacteria enumeration was done and microbiological classical methods were employed to characterize the microbial populations. PCR-DGGE was used to follow the biofilter community dynamics. Results obtained showed three stable levels of free bacteria distribution (lowest, intermediate and highest) and also that the biofilter was the main source of bacteria in the production system. PCR-DGGE data showed that the attached biofilter community presented seasonal and spatial segregation. The first 30 days were the most important for biofilter maturation: higher heterotrophic bacteria biomass loading and higher community diversity in the carriers. These results will be further analysed in conjunction with system operational data. This work was financially supported by the European Community (Project Raceways, COOP-CT/016869/2006-2008).

References: [1] Øiestad, V. (1999), Shallow raceways as a compact, resource-maximizing farming procedure for marine fish species, Aquaculture Research, 30, 831-840.

[2] Blancheton, J.P. (2000), Developments in recirculation systems for Mediterranean fish species, Aquaculture Engineering, 22, 17-31.

193

Information Systems about Materials for Architecture

F. Figueiredo 1

1 Department of Construction, Faculty of Architecture, University of Porto, Portugal.

What to build with is a question that has always remained with architects. The need for information and knowledge organization has always been felt. Recent transformations brought a stronger presence of these antic subjects. Architecture finds its balance at this present of stunning acceleration. The need of speeding up the projecting, without relinquishing quality, made man search for ways through specialization and interdisciplinarity. This work is intended to study if Information Systems (Inf. S.) about construction materials can take to high levels of interdisciplinarity at Architectural project and if the use of such instrument enables us to find a material that does what we want from it, instead of finding it based on what we know about it. Some bibliographic references gave clues about the context, complement and articulation at an Architectural level and at material’s generic subject; however, the most important references were Manzini [1], Sousa [2] and Nielsen [3]. An Instrument for characterization of S. Inf. about materials was built with the bibliographic support. The definition of the guidelines of this instrument was made simultaneously with the choice of the 17 study cases. The samples were used for testing the instrument and for base of later analysis, while studying deeply 4 of those cases. Reflections were made about different concepts over materials and their information, and it was understood which ones were the most relevant inter-confluences at materials definition in the process of Architectural project. The actual panorama of the on-line Inf. S. about materials was analyzed, through some characterization parameters, from the Instrument, and selected for the purposes. A series of potentialities in the study cases relating to the searching for materials through their properties were found and listed; the needs enounced by the authors in the ambit of Architectural project with the potentialities that were found in the studied Inf. S. were crossed. In short, it appears that we can say that the actual Inf. S. about construction materials can’t take us to a larger interdisciplinarity in the Architectural project yet. It happens because the different types of language and contents aren’t connected. Where it comes to searching materials by properties, it was found that some Inf. S. enable this and several other related features. However, there are limitations because they don’t merge into one search tool. The developed, used and validated Instrument can be applied to other studies or case studies in different or similar contexts. Also, the potentialities found during the analysis, together with the goal of associating different languages and contents, could contribute as a starting point for developing new or already existent Inf. S. about materials for Architecture. References: [1] Manzini, E. (1993), a matéria da invenção. Porto, Ed. Centro Português de design. [2] Sousa, J. (2005), “Convergência e não-linearidade no processo digital”. In Arquitectura e Vida (62), Lisboa, Ed. Loja da Imagem, pp. 28-32. [3] Nielsen, J. (2000), Designing Web Usability. Indianapolis, Ed. New Riders Publishing.

194

Osteogenic potential of adipose tissue-derived mesenchymal stem cells. Effect of medium composition and cell passage

C. Bessa Pereira1,2 and M.H. Fernandes2

1Faculty of Sciences, University of Porto, Portugal 2Faculty of Dental Medicine, University of Porto, Portugal,

Mesenchymal stem cells derived from adipose tissue have the potential to differentiate along different lineages, including the osteoblast phenotype. This characteristic makes them prospective useful for tissue engineering applications regarding the repair and regeneration of bone defects. The aim of this work is to evaluate the expansion and the osteogenic potential of serially-passaged adipose tissue-derived mesenchymal stem cells in different culture conditions. For comparison, stem cells derived from bone marrow, the most common source of osteogenic stem cells, are evaluated in parallel. Stem cells derived from subcutaneous adipose tissue and bone marrow are obtained from 3-months-old Wistar rats and cultured in -minimum essential medium containing 10% fetal bovine serum, antibiotics and 50 g/ml ascorbic acid, both in the absence (control medium) and in the presence of 10 nM dexamethasone. At 70-80% confluence, cells were subcultured in three experimental conditions: (i) control medium, (ii) control medium supplemented with 10mM Na-- glycerophosphate, and (iii) control medium supplemented with 10 mM Na-- glycerophosphate and 10 nM dexamethasone. Cell behaviour is evaluated at several passages for cell/viability proliferation and expression of osteogenic differentiation markers, by phase contrast microscopy, histochemistry and immunohistochemistry assays, flow cytometry and scanning electron microscopy. Preliminary results show that mesenchymal stem cells derived from rat adipose tissue present a high proliferation rate over several passages. Dexamethasone, a classic osteoblastic inducer, causes a significant reduction in cell growth rate with a simultaneous increase in the expression of alkaline phosphatase, an osteoblastic marker. In mineralising- favouring conditions, i.e., cultures supplemented with Na--glycerophosphate, mineralisation of the extracellular matrix is observed in selected conditions. Comparatively, mesenchymal stem cells derived from rat bone marrow present a higher expression of alkaline phosphatase and matrix mineralisation. In both, cell passage results in a gradual loss of these two osteoblastic parameters. More complete assessment of the cell populations regarding cell-cycle analysis and osteoblastic differentiation markers is being performed. The results of this work are expected to contribute to the characterisation of mesenchymal stem cells derived from adipose tissue regarding their proliferation and osteogenic differentiation patterns, over several passages, in order to assess their suitability for in vivo application in cell-based bone regeneration strategies. References: [1] Zuk, P.A., Zhu, M., Mizuno, H., et al (2001), Multilineage cells from human adipose tissue: implications for cell-based therapies, Tissue Engineering, 7, 211-228.

195

Potential of Equisetum arvense hydromethanolic extracts as osteoblastic growth promoters

C. Bessa Pereira1,2, L. Vieira3, M.A. Lopes4 and M.H. Fernandes2

1Faculty of Sciences, University of Porto, Portugal 2Faculty of Dental Medicine, University of Porto, Portugal 3Institute of Biomedical Sciences Abel Salazar, University of Porto, Portugal 4Faculty of Engineering, University of Porto, Portugal

Herbal medicines exert their beneficial effects through the additive or synergistic action of several chemical compounds acting at single or multiple target sites associated with a physiological process. Equisetum arvense (“horsetail”), a well known and wide-spread pteridophyte distributed in the northern hemisphere, contains a large variety of bioactive compounds including phenolics (flavonoids, phenolic acids, tannins), phytosterols, saponins and alkaloids. In vitro and in vivo studies provide some evidence of a positive impact of phenolic phytochemicals and phytosterols on bone metabolism [1]. In addition, horsetail has a high concentration of silicon, a mineral with an essential role in the bone formation process [2]. The aim of this work was to evaluate the effect of hydromethanolic extracts of E. arvense in the proliferation of human osteoblastic cells. Aiming at a potential application in bone regeneration, osteoblast cell behaviour was also assessed in colonized Bonelike® granules impregnated with the extracts. Bonelike® is a synthetic osteoconductive bone graft that mimics the inorganic bone matrix and has been successfully used in clinical bone regenerative applications [3]. MG63 osteoblast-like cells were cultured for 6 days in the presence of (i) hydromethanolic extracts of E. arvense (0.05 – 5%), (ii) silicon (0.005-0.5 ppm), and (iii) Bonelike® granules impregnated with the extracts. Cultures were assessed for cell viability/proliferation (MTT assay), throughout the incubation time. E. arvense extracts (0.05% and 0.1%) had a promoting effect on the proliferation of MG63 osteoblastic cells, between 29% and 54%. At 1%, the extracts caused generalized cell dead. Silicon, at 0.005 to 0.5 ppm, representative of the levels measured in the inducing extracts, caused an increase on cell proliferation around 20%, suggesting the presence of other bioactive compounds on the extracts. Bonelike® granules impregnated with the extracts allowed the adhesion and proliferation of MG63 osteoblastic cells. Based on these observations, it can be suggested that the association of Bonelike® with E. arvense extracts might be a promising approach for improving bone graft osteointegration, further enhancing the bone regenerative potential of this bone graft.

References:

[1] Habauzit, V. and Horcajada, M-N. (2008), Phenolic phytochemicals and bone, Phytochemical Reviews, in press, DOI 10.1007/s11101-007-9078-9. [2] Carlisle, E.M. (1997), Silicon, in O’Dell, B.L. and Sunde, R.A. (Eds.), “Handbook of nutritionally essential mineral elements”, Marcel Dekker Inc, New York, pp. 603-618. [3] Hussain, N., Lopes M.A., Maurício, A., Ali, N., Fernandes, M.A., Santos, J.D. (2007), Bonelike graft for bone regenerative application, in Ahmed, W. and Jackson, m. (Eds), “Surface engineered surgical tools and medical devices”, Springer Publications, New York.

196

Comparison of the in vivo bone forming activity of osteogenic- induced mesenchymal stem cells derived from adipose tissue and bone marrow

T. Ribeiro1,2, C. Bessa Pereira1,2, P. Gomes2 and M.H. Fernandes2

1Faculty of Sciences, University of Porto, Portugal 2Faculty of Dental Medicine, University of Porto, Portugal

Adipose tissue contains pluripotent stem cells able to osteogenic differentiation. These cells are ease to harvest in large amounts for autologous transplantation and present high proliferation rates for ex vivo expansion, making them attractive for bone tissue engineering approaches. The aim of this work is to compare the bone forming activity of mesenchymal stem cells from adipose tissue and bone marrow, after in vitro expansion over supportive scaffolds and subcutaneous implantation of the cell-material constructs in a rat model. As the vascularisation in the local environment plays a key role in the bone formation events, the angiogenesis response elicited in the different experimental conditions is also evaluated. Stem cells derived from subcutaneous adipose tissue and bone marrow are obtained from 3-months-old Wistar rats, following standard procedures. Culture is performed in - minimum essential medium containing 10% fetal bovine serum, antibiotics and 50 g/ml ascorbic acid, both in the absence and in the presence of 10 nM dexamethasone, a classic osteoblastic inducer. Cells of the first passage are cultured on clinically approved scaffolds in a combination of non-induced and osteogenic-induced conditions, following subcutaneous implantation in athymic nude mice. Prior to in vivo implantation, the cell- material constructs are characterized for cell growth and expression of osteoblastic differentiation markers. At defined time-points after implantation (typically, two and 12 weeks), the cell constructs with surrounding tissue are harvested, along with selected organs, and subject to histological evaluation regarding the expression of bone matrix proteins, bone forming activity and angiogenesis response. Preliminary results show that mesenchymal stem cells derived from rat adipose tissue present a high proliferation rate over several scaffolds, both ceramic-based and polymeric materials. Dexamethasone induces significant effects on cell growth rate and osteoblastic differentiation profile. At the same experimental conditions, bone marrow-derived mesenchymal stem cells appear to exhibit a higher osteogenic in vitro potential. In vivo studies are in course. The forthcoming results are expected to contribute to a more complete characterization of adipose tissue-derived mesenchymal stem cells regarding their suitability for tissue engineering applications in the repair or replacement of diseased and/or damaged bone tissue.

References:

[1] Zuk, P.A., Zhu, M., Mizuno, H., et al (2001), Multilineage cells from human adipose tissue: implications for cell-based therapies, Tissue Engineering, 7, 211-228.

197

Taxonomy from rural diffuse to urban industrial diffuse

André Chaves1

1 Faculty of Architecture, University of Porto, Portugal.

This work develops the relationship and the transition between the rural and urban diffuse industrial space, according to a system of mathematical classification, which is based on a case study in the Middle Ave. This research seeks to highlight the importance of the recent issue of urbanization diffuse addressed in the territorial planning along with the secular forms of ownership of land which are the rural areas. This space is important to diffuse urbanization because it is a background to its origins and to its cultural and material support. The decoding of both spaces was systematized according to a principle of structuring that identifies the following different parts: biophysical components, infrastructure, property, occupation and building. These parts were sorted in an empirical way, where the biophysical components constitute the beginning and building the end of a process. Each of these parts branches on a typological and material analysis. Initially begins by examining the rural diffuse space and, in a second phase, the urban industrial diffuse space as a result of a process called recombination of the different parts referred as related to rural diffuse space. The recombination is formed by methods of constant, addition, subtraction, and mutation of the rural diffuse. From this typological and material study was created a sequenced diagram in tree form (table 1). Here were exposed, in different sequences, all constituents of the various parts identified and set as an empirical mode. A conclusive summary of the relationship, in terms of taxonomic, of rural and urban diffuses spaces may be explained as this: the rural diffuse space as a concept identified as TSA - Topological Space Appropriation, auto recycled and auto sufficient in terms of a local cultural land use; the urban industrial space as a concept identified as PSA – Predefined Space Appropriation, continuous consume grow in terms of an influenced “globalized” imported cultural urban facts.

Table 1: Syntheses of the space taxonomy from the rural diffuse to the urban industrial diffuse.

198

Tetracyclines modulation of the osteogenic differentiation – in vitro evaluation

I. Côrte-Real 1, A. L. Marinho 1, P. S. Gomes 1, J. Rodrigues 1, M. H. Fernandes 1

1 Laboratório de Farmacologia e Biocompatibilidade Celular, Faculty of Dental Medicine, University of Porto, Portugal.

Tetracyclines are broad-spectrum antimicrobial agents that are characterized as modulators of several biological effects in cellular mechanisms, unrelated to the inhibition of ribosomal protein synthesis – their principal antibacterial mechanism of action [1]. High chelation activity with cations may account for several of the non-antimicrobial properties, at the same time that accounts for the high affinity to mineralized tissues, namely teeth and bone [2]. Although the established affinity, little is known about the direct or indirect modulation of bone metabolism by tetracyclines. The aim of this work was to evaluate the proliferation and differentiation events of osteoblastic-induced human bone marrow cells, in the presence of therapeutic concentrations of doxycycline or minocycline (semi-synthetic tetracyclines). Human osteoblastic cells were grown for 35 days, in the presence of doxycycline (1-25 µg/ml) or minocycline (1-50 µg/ml), on the surface of standard tissue culture plates. Cell cultures were evaluated for proliferation and differentiation events. Both pharmacological agents, at low concentrations (1µg/ml), reported an increased cell proliferation. After an initial lag phase, the increased cell growth was verified in association with expression of high levels of alkaline phosphatase and a proportional increased amount of mineralized extracellular matrix. Exposure to 5 µg/ml of doxycycline induced similar results, while the same concentration of minocycline impaired osteoblastic function. Higher levels of both tetracyclines greatly impaired cell proliferation and function in a dose-dependent way. Currently, characterization of tetracyclines’ influence during the in vitro osteogenic development is being evaluated by flow cytometric analysis of the stromal precursor cell marker STRO-1 and alkaline phosphatase expression, by osteoblastic-derived human bone marrow cells. This double staining allows to differentiate between four STRO-1/alkaline phosphatase subsets, which correspond to specific osteogenic differentiation stages. Overall, the treatment of human osteoblastic cells with low concentrations of doxycycline or minocycline induced cell proliferation without impairing functional activity, analyzed by alkaline phosphatase activity and extent of matrix mineralization. Its therapeutic usage might combine a targeted antimicrobial activity into the bone tissue with the induction of osteoblastic proliferation, without interfering with cell-specific biological activity. Flow cytometry analysis will contribute to evaluate tetracyclines influence in cell behavior, at different stages of the osteogenic differentiation. References: [1] Nelson, M. (1998), Chemical and Biological Dynamics of Tetracyclines. Adv Dent Res, 12, 5- 11. [2] White, J., Pearce, F. (1982), Characterization of chlortetracycline as a calcium ionophore. Biochemistry, 21, 6309-6312.

199

Study and design of precast reinforced concrete box-culverts under high embankments

Daniel Pinto1

1 Department of Civil Engineering, Faculty of Engineering, University of Porto, Portugal.

Precast reinforced concrete box-culverts (BC) are often used as systems for crossing highway embankments (materializing hydraulic and pedestrian passages), [1]. Although this is a simple role, the loadings applied to these structures are rather complex. Due to soil-structure interaction effects, the state of stress on the culvert depends on the stiffness of both the structure and the backfill, which is not usually contemplated in the simplified design methods that assumed that the load applied to the top slab correspond to the geostatic load. The applied soil pressure is also governed by the BC geometry, the soil properties and the method and quality of construction [1-4]. A numerical study on reinforced concrete box-culverts behaviour was developed on a precast commercial BC design for a 10m high embankment. The numerical analyses were performed using a finite element code, in the DIANA-9.2 program, capable of considering the nonlinear behaviour of both the soil and the reinforced concrete structure and enabling the consideration of all the construction stages. New parametric analyses, beyond those which were accomplished in previous studies, [2, 3] were developed for the identification of the main parameters influencing the interaction mechanism and to evaluate the BC structural performance up to failure. These parametric analyses were enclosed on four principal analyses groups, like changing the soil characteristics, the concrete characteristics, the reinforced quantity and changing the BC geometry. The influence of the BC nonlinear behaviour on the interaction mechanism, both in service and ultimate state conditions, was analyzed and discussed. The main results show that the soil characteristics have an important influence in the BC behaviour and the other parameters affect the performance up to failure. The principal cause of failure is the concrete crushing on the interior corners on the BC walls. As concluding remarks, it is considered that the soil pressures and the BC nonlinear behavior are directly related and should be reflected in the design stage in order to achieve a more rational and economical design.

[1] Bennett, R., Wood, S., Drumm, E., and Rainwater, N. (2005), Vertical Loads on Concrete Box Culverts under High Embankments, Journal of Bridge Engineering, 10(6), 643-649. [2] Figueiras, J., Pimentel, M., Costa, P. (2006), Análise do Comportamento e Verificação de Dimensionamento de Box-Culverts – Relatório Síntese, LABEST-FEUP. [3] Sang, Pedro A.S. (2000), Comportamento e Modelação Numérica de Estruturas Enterradas de Betão, Dissertação de Mestrado em Estruturas de Engenharia Civil, FEUP. [4] Kim, K., and Yoo, C. (2005), Design Loading on Deeply Buried Box Culverts, Journal of Geotechnical and Geoenvironmental Engineering, 13(1), 20-27.

200

Properties of nanostructured materials

C. P. Arieira, F. Silva and C. M. Pereira

Department of Chemistry, Faculty of Science, University of Porto, Portugal. Rua do Campo Alegre, 687, 4169-007 Porto, Portugal

Synthesis and characterization of gold nanoparticles and their self-assembly on modified surface for electrochemical studies and sensor applications. Several synthetic methods were chosen all involving chemical reduction of the metal salt in the solution. The particles prepared include gold nanoparticles functionalized with different ligands such as citrate, lysine, cytosine, and different sugares, glucose, sucrose, sorbitol, manitol and dulcitol.

0,7 Absorvance = f (wavelenght(nm))

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Sucro se

0,4 Manitol So r bit o l

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Absorbance

0,2

0,1

0 400 450 500 550 600 650 700 Wavelenght(nm)

Figure 1 shows the evolution of UV-vis spectra during the formation of gold nanoparticles using the various sugares as reducting agents at λmax~523nm.

The experimental results will be analyzed in order to characterize the gold nanoparticles for Voltammetric Analysis subsequent.

Acknowledgments: Financial support from FCT (POCI 2010) and FEDER for the project POCI/QUI/57679/2004 is gratefully acknowledged. References: [1] J. Chemistry Education, Synthesis of Colloidal Gold, (2004) 81, 544A. [2] S. Panigrahi, S. Kundu, S. K. Ghosh, S. Nath, T. Pal, Sugar assisted evolution of mono- and bimetallic nanoparticles, 264 (2005) 133-138.

201

Influence of egg yolk and aromatizing compounds on the volatile profile and on sensory characteristics of bakery cream

B. Ramos 1, O. Pinho2, and I.M.P.L.V.O. Ferreira1

1 REQUIMTE- Serviço de Bromatologia, Faculdade de Farmácia da Universidade do Porto, Rua Anibal Cunha 164, 4099-030 Porto; Portugal 2 Faculdade de Ciências da Nutrição e Alimentação, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto – Portugal

Egg bakery cream (EBC) is widely appreciated in bakery products. Traditionally, this cream was prepared with egg yolk. Two different methods could be used, one prepared with sugar and water heated until bland caramelization, added of egg yolk, vanilline and farine (EBC1), and the other, prepared by careful heating of milk, sugar and farine added of egg yolk and vanilline (EBC2). Presently, bakery cream substitutes (BCS), containing egg aromatizing compounds, and prepared with cold water are widely used in bakery products. The flavour of traditional EBC is the result of the aroma compounds produced by thermal reactions during heating of egg yolk, sugar, farine, and other ingredients. Lipid oxidation and Maillard reactions are the most important pathways leading to egg products odorants. Previous studies of the aromatic compounds in baked products have mainly focused on bread and bread-making steps [1]. Aditionally, volatile compounds that contribute to the aroma in the amounts present in heated egg yolk were identified [2]. To our knowledge, no study on the odorants present on EBC and on BCS has been published. The aim of the present investigation was to determine the volatile compounds that can contribute to the aroma of EBCs and compare with the volatile fraction of a bakery cream prepared by a similar process but without adding eggs (NBC), the volatile compounds of a BCS commercially available and bakery creams collected from bakery products randomly purchased from the marked. Volatile profile of bakery cream was obtained by solid phase microextraction with CAR/PDMS fiber coupled with GC/MS. Chemometric analysis of the volatile data was also used to predict bakery cream origin. Additionally, the sensory descriptive profile of the different types of bakery creams was elaborated. Ninety two volatiles were identified after extraction, including, aldehydes, ketones, esters, aromatic compounds, alcohols, hydrocarbons, furans and acids. The compounds were identified by comparing the experimental spectra with those of the Nist’ 98 data bank. EBC1 and EBC2 presented significantly different volatile profile. As expected EBC1 presented high variety of volatiles owing to the Maillard reactions resulting from sugar heating during a longer period. The discriminante analysis applied to SPME-GC/MS data indicated differences between NBC, EBC1 and EBC2, and similarities between EBC1, BCS and bakery creams collected from bakery products. The function thus obtained was able to correctly classify all the samples according to bakery cream origin. However, different sensory descriptive profiles were obtained for bakery creams of different origin.

References: [1] Pozo-Bayón, M.A. (2007) J. Agric. Food Chem. (55) 1418- 1426. [2] Cerny, C.; Guntz R. (2004) Eur. Food Res. Technol. (219) 452-454.

202

Evaluation of genetic and chemical diversity in Portuguese populations of Phaseolus vulgaris L.

R. Coelho1, M. A. Faria2, A. Barata3, M.B.P.P. Oliveira2 and E. Nunes1

1 CIBIO – Laboratory of Vegetal Genetics, Faculty of Sciences, Universitiy of Porto, Campus Agrário de Vairão, Rua Padre Armando Quintas, Vairão, 4485-661 Vila do Conde 2 REQUIMTE – Serviço de Bromatologia, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030 Porto 3 BPGV – Portuguese Bank of Vegetal Germoplasm, DRAP-N, Quinta de S. José, S. Pedro de Merelim, 4700 Braga

The common bean (Phaseolus vulgaris L.) is the most consumed legume in the world. In Portugal, the geographic isolation of the region surrounded by the mountainous barrier of Peneda-Gerês, Barroso and Marão it is thought to have the safeguard of a large number of adapted bean populations. In the present study, 20 Portuguese accessions, collected by the Portuguese Genebank (BPGV) from the referred region were evaluated in two locations - Braga and Vieira do Minho - for chemical (protein content) and genetic diversity (microsatellite DNA). Crude protein content was determined using the Kjeldahl method. The molecular characterization was achieved by analyzing six microsatellite loci described by Gaitán-Sólis [1]. Genetic data was analysed using the programs Identity, Microsat, Phylip and Tree View. Significant differences in crude protein content were found among locations, accessions, and location by accession interactions. The average crude protein content was higher in Vieira do Minho than in Braga (29.31% and 27.79%, respectively). The 6 microsatellite loci used for molecular characterization contributed to differentiate most of the accessions and the 20 accessions were grouped in terms of genetic proximity in three main clusters. The studied collection of common beans presented genetic variability on crude protein content among and within accessions potentially useful in plant breeding programs. Further detailed studies are warranted, to fully understand the potential of the germplasm maintained at BPGV.

[1] Gaitán-Solís E, Duque M.C., Edwards K.J., Thome J. (2002), Microsatellite Repeats in Common Bean (Phaseolus vulgaris): Isolation, Characterization, and Cross-Species Amplification in Phaseolus ssp., Crop Science, 42, 2128-2136.

203

The metamorphosis of Aleixo towers

A. Lima 1

1 Department of Architecture, Faculty of Architecture, University of Porto, Portugal.

Aleixo is a social neighborhood that consists in a unique experience in the city of Porto. The first drawings of the project date of 1968 and its construction was concluded in 1976. It was designed for the reaccomodation of the families to displace from Ribeira-Barredo during its renovation. The complex is composed of five towers with 13 floors each, whose 320 dwellings (two T2, two T3 and one T4 by floor) are organized around an open space that ventilates and illuminates all the central part of the tower, where the vertical accesses are located and where the kitchens, bathrooms and entrances turn to. It is clear the gradual degradation of the construction, as well as the social degradation of this complex through the years, while the polemic dicotomy demolition/non demolition has been a constant. The project of the transformation of the towers is based on the study Plus, les grands ensembles de logements. Territoires d´exception that questions the public program lead in France of demolishing many communal housing complexes built on the 1960s and 70s to change their negative image. At the same time, this is also a stand-by solution for the Aleixo social neighbouhood. The proposed project goes along with this manifest: “We think that demolishing the existing would be aberrant and instead of that, transforming it would permit a cheaper, effective and more quality answer to the existing necessities. The architecture of these “grands ensembles” are often obsolete and generally in inadequacy with the actual needs of the inhabitant. Yet we are convinced by the latent potential of these architectures. The conservation and pertinent analysis of the structural, geographic and spatial qualities of these important buildings could lead to a dramatic transformation following radical objectives. These are to allow for the size of units to be twice as generous, filled with natural light, allow for different and non typical typologies of flats. To offer improved services and usages and to consider the quality of the interior and common spaces as a priority over the urban gesture. These are the contemporary aims.” [1] The proposal of intervention in the towers can be seen as an example of a inovating reutilisation that adapts to the existing (who knows the sketch of a new heritage?) and that could ilustrate and be monitorised by the evolution of the urban policies.

References: [1] Druot, Frédéric, Lacaton, Anne and Vassal, Jean – Phillipe, Plus, les grands ensembles de logements. Territoires d´exception, Ministère de la Communication, Direction de Lárchitecture et du Patrimoine, p.9

204

Modulation of cytochrome P450 and oxidative stress by rooibos (Aspalathus linearis) infusion intake

D. Teixeira1,2, A. Marinho2, C. Meneses2, I. Ferreira2, J. Sampaio2, M. A. Lima2, V. Mendes1,2, A. Faria1, R. Monteiro1,2, C. Calhau1

1Department of Biochemistry (U38/FCT), Faculty of Medicine, University of Porto, Portugal. 2Faculty of Nutrition and Food Sciences, University of Porto, Portugal.

Cytochrome P450 enzymes (CYP) represent a large family of proteins involved in the metabolism of drugs and other xenobiotics, as well as some endogenous substrates. Drug interactions can frequently arise when drugs are co-administered and one drug modifies the metabolic clearance of the second drug by inhibition or induction of a specific CYP enzyme. It has been described that different natural products can affect activity of CYP enzymes. Thus, concomitant drug and food intake creates the opportunity for interactions that may change the kinetics and resulting effectiveness or toxicity of a drug. A notable example of this type of interaction is grapefruit juice and St. John’s Wort (1), which results in a strong inhibition of CYP3A activity (2,3). Rooibos (Aspalathus linearis) infusion is becoming a popular drink, as it is claimed to protect health through antioxidant activity. Thus, we thought of interest to test the effect of rooibos infusion intake on hepatic CYP concentration and activity and on oxidative stress. To do so, we treated CD1 mice for 2 weeks with rooibos infusion (n=6) and included an appropriate control group (waterdrinking, n=6). After treatment, pentobarbital (30 mg/kg b.w., i.p.)-induced sleeping time was determined to estimate CYP activity, since this hypnotic drug is inactivated by CYP, particularly isoenzyme 2E1. Afterwards, animals were sacrificed and livers removed, homogenized and used to determine total hepatic CYP450 content by spectrophotometry at 450 nm in reduced hepatic microsomes saturated with carbon monoxide. As an oxidative stress biomarker, thiobarbituric acid-reactive substances were determined to estimate lipid peroxidation. Pentobarbital-induced sleeping time tended to decrease after rooibos infusion ingestion, total CYP450 content was significantly increased and this was accompanied by a higher hepatic level of TBARS. In conclusion, our results underscore important toxico- and pharmacokinetic alterations resulting from rooibos infusion consumption involving CYP, enhancing or decreasing the bioactivity of its substrates. This is particularly important if exposure to procarcinogens occurs simultaneously with rooibos consumption, as their activation is likely to be increased. Whatever the case, lipid peroxidation seems to be higher, probably due to CYP stimulation, what can be per se related to detrimental effects.

(1) Sparreboom A, Cox MC, Acharya MR, Figg WD: Herbal remedies in the United States: potential adverse interactions with anticancer agents. J Clin Oncol 2004;22:2489-2503. (2) Obach RS: Inhibition of human cytochrome P450 enzymes by constituents of St. John's Wort, an herbal preparation used in the treatment of depression. J Pharmacol Exp Ther 2000;294:88- 95. (3) Wang Z, Gorski JC, Hamman MA, Huang SM, Lesko LJ, Hall SD: The effects of St John's wort (Hypericum perforatum) on human cytochrome P450 activity. Clin Pharmacol Ther 2001;70:317-326.

205

Screening for novel CDH1 inactivating mechanisms in Familial Gastric Cancer

P. Inácio1, H. Pinheiro1, J. Carvalho1, S. Sousa1, R. Seruca1,2, C. Oliveira1,2

1 1.IPATIMUP, Porto, Portugal 2 Department of Pathology, Faculty of Medicine, University of Porto, Portugal

Gastric cancer exists mainly in its sporadic form but familial aggregation of the disease occurs in 10% of the cases. Despite all efforts to determine the genetic basis of familial gastric cancer, a single gene, CDH1, has been identified with a causative role in Hereditary Diffuse Gastric Cancer (HDGC) and Familial Diffuse Gastric Cancer (FDGC) [1]. Two thirds of HDGC and 90% of FDGC families displaying gastric tumours of the diffuse histotype remain genetically unexplained. Since no other candidate gene has been described in these families, but the hereditary pattern of inheritance and the tumour’s histotype remains the same as the families carrying CDH1 mutations, our aim is to identify new CDH1 inactivation mechanisms that may underline the cancer associated familial history observed in these patients. We screened DNA extracted from peripheral blood lymphocytes (PBLs) from 38 probands by PCR/Sequencing for two ESTs in CDH1 intron 2 that may represent putative regulatory regions of the CDH1 expression. Moreover, we analysed PBLs DNA from 62 probands for germline CDH1 promoter methylation by bisulphite DNA treatment and MS- PCR/Sequencing, since germline epimutations have been described as disease cause in other cancer associated syndromes. We identified a new sequence variant (A>T) at position 27.109 of intron 2, in 22/38 (57.9%) family probands. No other alterations were found at the screened regions. Hypermethylation of the promoter CpG island 3 of the CDH1 gene was found in PBLs DNA from two probands of Portuguese families. The screening of normal and tumour tissue from probands and first degree relatives will confirm the possibility of this being a heritable germline epimutation. The confirmation of such results will disclose a new inactivating mechanism of CDH1 in families displaying gastric cancer.

References:

[1] Guilford P., Hopkins J., Harraway J., McLeod M., Ngahiraka M., Harawira P., Taite H., Scoular R., Miller A., Reeve A.E. (1998) E-cadherin germline mutations in familial gastric cancer. Nature 392, 402-404

206

The influence of interpersonal similarity in social categorization

D. Teixeira 1, F. Gonçalves 1, I. Lourenço 1 and S. Duarte 1

1 Faculty of Psychology and Educational Sciences, University of Porto, Portugal (students)

The main goal of this study was to articulate two theoretical concepts generally regarded as separated in social psychological research: the interpersonal similarity (related to a psychological or intra-individual level of comprehension) and the social categorization (more related to the social or inter-grupal level). The present study followed the premises of the Subjective Group Dynamics model (e.g. Marques, Abrams, Páez & Hogg, 2001) which proposes that people are motivated to ensure the validity of a subjective sense of reality (that is defined and shared by their groups) as a way to defend their own identity. From that starting point, this work studied the impact of interpersonal similarity on social categorization, as a way to reduce or implement a subjective identification with a social group. On a judgmental activity, subjects were asked to evaluate a number of fictional targets (though presented as real), with the interpersonal similarity and the group membership being manipulated across conditions. Namely, subject being very similar to, somewhat similar or very different from most targets (interpersonal similarity) versus in-group and out-group targets (targets´ group membership). It was found that interpersonal similarity had the most impact on social judgments, with subjects being more positive toward their own equals. However, amidst these results there was an important phenomenon in regard. When subjects were very different from most targets of their own group, they lessened their identification with their group and regarded best the somewhat similar targets. The main point is that when subjects found themselves as very different from most members of their own social group they gave more relevance to another kind of social categorization: being an atypical or marginal member of the in-group. As an alternative base of social inclusion, they increased identification with in-group members that were similarly “stigmatized” (Major and Eccleston, 2005).

References: [1] Marques, J. M., Abrams, D., Paez, D., & Hogg, M. A. (2001), Social categorization, social identification, and rejection of deviant group members, In Hogg, M. A. & Tindale, R. S. (Eds.), Blackwell handbook of social psychology: Group processes (Vol. 3, pp. 400-424). Oxford, UK: Blackwell Publishers. [2] Major, B., Eccleston, C. P. (2005), Stigma and social exclusion, In Abrams, D., Hogg, M. A., & Marques, J. M. (Eds.), The social psychology of inclusion and exclusion (pp. 63-87). East Sussex, UK: Psychology Press.

207

Genetic characterization by SSR of six Olea europaea regional varieties for authenticity evaluation of olive oils

J. Lopes1, R. Coelho2 , M. A. Faria1, E. Nunes2 and M.B.P.P Oliveira1

1REQUIMTE, Serviço de Bromatologia, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha, 164, 4099-030 Porto, Portugal. 2 CIBIO, Faculdade de Ciências da Universidade do Porto, Campus Agrário de Vairão, Rua do Monte-Castro, Vairão, 4480-661 Vila do Conde

Olive cultivation plays an important role in rural economy of certain regions as Trás-os- Montes. The importance attributed to this cultivar is expressed in the number of regional varieties that constitutes local germplasm collections. In order to assist germplasm management and to develop markers for olive oil varietal authentication genotyping the referred varieties is of crucial importance. Several methodologies have been used to genotype and evaluate the relationships among cultivars of olive tree, including isozymes, RFLP, RAPD, ISSR, SSR and SNPs. SSR are the markers of choice since they are highly polymorphic, co-dominant and relatively easy to use. In this work we have studied the six most relevant regional varieties (Cobrançosa, Verdeal Transmontana, Madural, Santulhana, Redondal e Negrinha) at the molecular level using 15 selected SSR loci [1-3]. Genomic ADN was obtained from silica gel dried leaves using both a CTAB classical method and a commercially available extraction kit. Results include the construction of a database for further olive oil authenticity studies and the evaluation of the genetic relationships amongst the varieties. The chosen markers permitted the complete discrimination of the studied varieties with a probability of identity of 1,52×10-5. Genetic relationships amongst the varieties are graphically represented in Fig. 1.

88 Fig. 1. Dendrogram showing the genetic relations 78

68 of the 6 studied varieties

58

48

Linkage Distance Linkage 38

28

18 REDONDAL COBRANÇOSA SANTULHANA NEGRINHA VERDEAL TRANSMONTANA MADURAL

[1] Sefc, K.M., Lopes, M.S., Mendonça, D., Santos, M.R., Machado, M.L.C., Machado, A.C. (2000), Identification of microsatellite loci in olive (Olea europaea) and their characterization in Italian and Iberian olive trees ,Molecular Ecology, 9, 1171-1193. [2] Cipriani, G., Marrazzo, M.T., Marconi, R., Cimato, A., Testolin, R. (2002), Microsatellite markers isolated in olive (Olea europaea L.) are suitable for individual fingerprinting and reveal polymorphism within ancient cultivars ,Theor Appl Genet., 104, 223-228. [3] Rallo, P., Dorado, G., Martín, A. (2000), Development of simple sequence repeats (SSRs) in olive tree (Olea europaea L.), Theor. Appl. Genet., 101, 984-989.

208

Programa Social

10h00 - Ponto de Encontro na Reitoria Passeio de Carro Eléctrico

11h30 Visita ao Palácio da Bolsa

12h30 Visita às Caves Calem

13h15 Almoço Livre

15h30 - Ponto de Encontro na Casa do Infante

16h00 Visita à Sé do Porto Visita à Estação de S. Bento

Fim da visita junto à Casa da Música

209

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