SANKO University Innovation in Medicine Summit-4, 11-13 Oct 2018, Gaziantep, Turkey.
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SANKO University Innovation in Medicine Summit-4, 11-13 Oct 2018, Gaziantep, Turkey.
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SANKO University Innovation in Medicine Summit-4, 11-13 Oct 2018, Gaziantep, Turkey.
Honorary President Prof. Dr. Güner DAĞLI SANKO University Rector Surrogate
Honorary Board Members
Prof. Mehmet Metin Bayram Vice Rector / SANKO University Prof. Salih Murat AKKIN Dean / SANKO Medical School Prof. Ayşen BAYRAM Director / SANKO University Institute of Health Sciences Dr. Yusuf Ziya YILDIRIM Genel Secretary / SANKO University
Congress President Prof. Eyüp İlker SAYGILI Assoc. Prof. Zafer ÇETİN
Congress Secretary Assist. Prof. Necla BENLİER Assist. Prof. Burçin ALTINBAŞ Assist. Prof. Duygu GÖK YURTSEVEN
Execution Commite Members
Eda ÖĞÜT Nilay UÇAR
Ebru YAPAN Kadir CENGİZ
Begüm TEKİN Burcu DERVİŞOĞLU
Hannan KELAHMETOĞLU
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International Sciencetific Commite
Prof. Aldo TOMASI (Universita Degli Modena Departmant of Diagnosis Medicine)
Prof. Atanas ATANASOV (Institute of Genetics and Animal Breeding PAS, Warsaw, Poland, Head of Molecular Biology Department)
Prof. Chryssostomos CHATGILIALOGLU (National Centre of Scientific Research "Demokritos", Athens, Greece Institute of Nanoscience and Nanotechnology)
Prof. Ozlem SORAN (Pittsburg University, Sanko University, Gaziantep, Turkey, Cardiology Department)
Prof. Seyed Mohammad NABAVI (Baqiyatallah University Medical Sciences, Iran, Applied Biotechnology Research Center)
Prof. Utkan DEMIRCI
(Standford Medical School, USA)
Assoc. Prof. Alexandros GEORGAKILAS (National Technical University of Athens, DNA Damageand Repair Laboratory)
Assoc. Prof. Oral CENK AKTAŞ (Christian Albrechts University, Institute of Materials Science, Saarland University, Crc Biomaterials, Germany)
Assist. Prof. Kaan YILANCIOĞLU (Üsküdar University)
Assist. Prof. Oktay KAPLAN (Abdullah Gül University)
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Dr. Şule Beyhan OZDAS (---)
Dr. Zacharenia NIKITAKI (National Technical University of Athens, DNA Damage and Repair Laboratory Biomedical Informatics)
National Sciencetific Commite
Prof. Abdullah OLGUN (İstinye University, Faculty of Pharmacy)
Prof. Ayse Munife NEYAL (Sanko University Medical School, Department of Neurology)
Prof. Belma KONUKLUGİL (Ankara University, Faculty of Pharmacy)
Prof. Burhan ATEŞ (İnönü University, Faculty of Science And Literatury Chemistry Department)
Prof. Cengizhan ÖZTÜRK (BosphorusUniversity, Institute of Biomedical Engineering)
Prof. Dildar KONUKOĞLU (İstanbul University Cerrahpaşa Medical School, Department of Medical Biochemistry)
Prof. İsmet YILMAZ (İnönü University, Chemistry Department of Science and Literature Faculty)
Prof. Kadir BATCIOĞLU (İnönü University, Faculty of Pharmacy Basic Pharmacy Sciences)
Prof. Mehmet Eyyuphan YAKINCI (İskenderun Technical University, Head of Department of Metallurgy and Materials Engineering)
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Prof. Mehmet YILMAZ (Sanko University Medical School, Department of Internal Medicine)
Prof. Melek ÖZTÜRK SEZGİN (İstanbul University Cerrahpaşa Medical School, Department of Medical Biology)
Prof. Mustafa BAYRAM (Gaziantep University, Head of Food Technology Department)
Prof. Münevver SÖKMEN (Konya Food and Agriculture University)
Prof. Nezih HEKİM (Cumhuriyet University Medical School, Department of Biochemistry)
Prof. Oya IŞIK (Çukurova University, Faculty of Fisheries Department of Marine Biology)
Prof. Sibel BERKER KARAÜZÜM (Akdeniz University Medical School, Department of Medical Biology and Genetics)
Prof. Turgut ULUTİN (İstanbul University Cerrahpaşa Medical School, Department of Medical Biology)
Assoc. Prof. Meral YÜKSEL (Marmara University, Vocational School of Health Services/Department of Medical Services and Techniques)
Assoc. Prof. Selin SAYIN (İskenderun Technical University, Faculty of Marine Sciences and Technology)
Assoc. Prof. Tuğba TAŞKIN TOK (Gaziantep University, Chemistry Department of Science and Literature Faculty)
Assist. Prof. Gökhan GÖRGİŞEN (Van Yüzüncü Yıl University, School of Medicine, Department of Medical Biology)
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Assist. Prof. Mehmet Şerif CANSEVER (Namık Kemal University)
Assist. Prof. Muhammed KARAMAN (Kilis 7 Aralık University, Chemistry Department of Science and Literature Faculty)
Assist. Prof. Süray PEHLİVANOĞLU (Konya Necmettin Erbakan University, Molecular Biology and Genetics Department of Science and Literature Faculty)
Assoc. Dr. Meral YÜKSEL (Marmara University Health Services Vocational School / Department of Medical Services and Techniques)
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SPEAKERS:
• Prof. Banu ONARAL, PhD
H. H. Sun Professor of Biomedical Engineering and Electrical Engineering at Drexel University, Philadelphia, PA, USA.
Banu Onaral is H. H. Sun Professor of Biomedical Engineering and Electrical Engineering at Drexel University, Philadelphia, PA. She received her BS and MS in Electrical Engineering from Boğaziçi University, Istanbul, Turkey, in 1973 and 1974, respectively, and earned her PhD in Biomedical Engineering from the University of Pennsylvania in 1978. Dr. Onaral joined the faculty of the Department of Electrical and Computer Engineering and the Biomedical Engineering and Science Institute at Drexel University in 1981. Starting in 1995, she led the strategic planning to transform the Biomedical Engineering and Science Institute into a university-level interdisciplinary school. She served as the Founding Director of the School of Biomedical Engineering, Science and Health Systems since its creation in 1997 to 2014. She currently leads the ‘Global Innovation Partnerships’ initiative as Senior Presidential Advisor at Drexel University.
• Prof. Utkan DEMİRCİ, PhD
Professor with tenure at the Stanford University School of Medicine, Stanford, CA, USA.
Dr. Utkan Demirci is a Full Professor with tenure at the Stanford University School of Medicine, Palo Alto, CA. Before Stanford, he was an Associate Professor of Medicine and Health Sciences and Technology at the Harvard Medical School (HMS) and Massachusetts Institute of Technology (MIT) Health Sciences and Technology (HST) division. He received his bachelor’s degree (summa cum laude) from the University of Michigan, Ann Arbor, his master’s degrees in Electrical Engineering in 2001 and in Management Science and Engineering in 2005, and his doctorate in Electrical Engineering in 2005 all from Stanford University. Dr. Demirci received IEEE EMBS Early Career Award; IEEE EMBS Translational Science Award; NSF CAREER Award; Coulter Foundation Early Career Award; HMS- Young Investigator Award; and Chinese National Science Foundation International Young Scientist Award. In 2006, he was selected to TR-35 as one of the world’s top 35 young innovators under the age of 35 by the MIT Technology Review.
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• Prof. Cengizhan ÖZTÜRK, PhD
Professor at Bogaziçi University, İstanbul, TURKEY
Dr. Cengizhan Ozturk has been leading a multidisciplinary research team at the Bogazici University: The Micro-System Based Medical Device Development Laboratory.It was more firmly established on January 2008, when it was funded through three-way project consortium by the State Planning Organization, with the official name of Center for Life Sciences and Technologies.(http://lifesci.boun.edu.tr) Original LİfeSci modules (DeviceLab, System Biology and Vivarium) have been expanded to include two new modules: a) Smart Drugs, b) Computational Biology and Bioinformatics. The Center is supported by over 40 academicians from 10 different departments. Vivarium is now fully operational, and the 250 m2 Clean Room, combining unique MEMS and medical device production capabilities under one roof, certified in Autumn of 2011. The overall earmarked budget for this ongoing center project is now around 9.500.000 $. In May 2011, Dr. Ozturk has been named the Director of the Center for Life Sciences and Technologies.
• Prof. Nesrin ÇOBANOĞLU
Istanbul University, Cerrahpasa School of Medicine, İstanbul, TURKEY
Prof. Dr. Nesrin Cobanoglu has graduated from Istanbul University, Cerrahpasa School of Medicine. She completed her graduate studies in "Public Administration" and "History of Medicine, Deontology and Medical Ethics” to become an Associate Professor in 2005 and a Professor in 2011. She is the founder of the Department of History of Medicine and Medical Ethics in Baskent University. She is currently working as the chair of the Department of Medical Ethics and History of Medicine in Gazi University. She is giving doctoral courses in the Institute of Social Sciences in Ankara University (Faculty of Political Science). Thus, she continues her scientific career in both areas. She has been the president of “Women’s Studies Center” in Gazi University. She has many national and international publications and books. She worked on the ethics boards of many national and international scientific organizations. She has been invited to UN, WHO, UNESCO and the EU scientific meetings as the representative of Turkey. She is a member of Health Ministery Scientific Committee.
She has been the member of the administrative board of Turkish Medical Association and Ankara Medical Association (General Secretary). She is currently a member of Turkish Medical Association, Biopolitics International Organization (BIOS), BPW (Business and Professional Woman), Sociological Association, AIDS Prevention Society, Hacettepe University AIDS Treatment Center (HATAM), Public Management Professionals Association, the Philosophical Society of Turkey, Bioethics Association, International Association for Education in Ethics (IAEE), Association of Medical Ethics and Medical Law.
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• Prof. Yalda Shokoohinia
Iran / Kermanshah University of Medical Sciences
Yalda Shokoohinia has completed her Pharm D in 2005 and her PhD in 2011 both from Isfahan University of medical sciences. She started as an assistant professor in Kermanshah University of Medical Sciences in 2011 and awarded as associate professor in 2015. She has been the head of department of Pharmacognosy & Biotechnology from 2013 to 2015. She has beenalso head of continuous medical education of Kermanshah University of Medical Sciences from 2015. She has published more than 34 articles indexed in ISI web of science, Pubmed, Scopus , etc and has supervised more than 30 theses of Pharm D and MSc students.
• Assoc. Prof. Alexandros GEORGAKILAS, PhD
Associated Professor at National Technical University of Athens (NTUA), Athens, GREECE
Dr. Alexandros Georgakilas is currently an Associate Professor in the Department of Physics, School of Applied Mathematics and Physical Sciences, National Technical University of Athens (NTUA), Greece. At Brookhaven National Laboratory, USA, Dr. Georgakilas completed post-doctoral research as a research associate in the Biology Department from 2001-2003 under the supervision of Dr. Betsy Sutherland and developed his own research laboratory as an Associate Professor with tenure at East Carolina University until 2012. Overall, Dr. Georgakilas has been an active non-stop researcher in the field of Radiation Biology for more than twenty (20) years. He has also received awards such as the Young Investigator Travel Award from Radiation Research Society, Radiation Research Society SIT Award, ECU Thomas Harriot College Research Award, and the prestigious Terashima Award from Japan Radiation Research Society. His work at ECU as Principal Investigator (PI) has been funded by various sources like East Carolina University, NCI, NC Biotechnology Center and Union for International Cancer Control (UICC). His quality research work has been published in more than fifty (60) peer-reviewed high profile journals like Radiation Research, Cancer Letters, Cancer Research, Pharmacology Therapeutics, Free Radical Biology and Medicine, Journal of Cell Biology and Proceedings of National Academy of Sciences USA and more than 4000 citations.
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• Assoc. Prof. Erkan KAPLANOĞLU
Associated Professor at Marmara University, Research Professor at Tennessee State University (BARLab.)
Dr. Kaplanoglu completed his M.Sc. and Ph.D. thesis at Computer-Control Department of Marmara University, Institute for Graduate Studies in Pure and Applied Sciences, in 2000 and 2006, respectively. He worked as a research assistant professor at Department of Mechanical Eng. Vanderbilt University, Nashville,TN Faculty of Engineering from 2011 to 2012. In March 2014, Dr. Kaplanoglu became Associate Professor at Marmara University, Mechatronics Department. He has been awarded YGGDRASIL Award (Young Guest Doctoral Researchers in Norway) in 2010. In 2013, He introduced the idea and oversaw the establishment of a Biomechatronics& Assistive Technology Lab (BioMechatLab) at Marmara University that has since built a reputation and is ranked second in the country in terms of student involvement and research. The establishment of this lab has led to the collaborations with the departments of biomedical and medical at other partner higher education institutions. He is currently a visiting researcher at the College of Engineering of Tennessee State University., Nashville, TN. His current research focuses on the Biomechatronics Systems, Lower Limb Prosthesis and Wearable Rehabilitation Devices.
• Assoc. Prof. Cenk AKTAŞ, PhD
Associated Professor at Saarland University and Applied University of Kaiserslautern, Saarbrücken, GERMANY
Assoc. Prof. Cenk Aktas earned his BSc and MSc in Materials Science and Engineering from Middle East Technical University-Turkey and Christian-Albrechts University-Germany, respectively. He joined Leibniz Institute for New Materials (Leibniz-INM) in 2004. After completing his PhD with distinction (summa cum laude) he was appointed as the Deputy Head of CVD Research Division. Between 2010- 2015, Aktas acted as the Director of CVD/Biosurfaces Division at Leibniz-INM, which is situated in Saarbrücken/Germany. In addition to his academic duties (acting as senior instructor at Saarland University and Applied University of Kaiserslautern), he gained invaluable experiences at Leibniz-INM since it is a well-known scientific partner to national and international institutes and a provider of research and development for companies throughout the world. Aktas published more than 70 research papers and 10 patents in different fields (on nanomaterials and nanotechnology). He has several prestigious awards including Prof. Werner Petersen Award, Prof. Horst Hardt Award, Prof. Baki Komsuoglu Award, International Nanomedicine Foundation Award and etc.
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• Assoc. Prof. Mazhar ADLI, PhD
Associated Professor at Virginia University, Charlottesville, Virginia, USA.
The Adli lab is focused on understanding whole-genome level chromatin regulation during normal (Cell, 2013) and disease development. We are specifically focused on understanding aberrant chromatin regulation in cancer progression and chemotherapy resitance Cancer (Cancer Cell, 2012; Nature, 2012, Nature 2011). We are using Genomics and Epigenomics profiling tools such as ChIP-Seq,ATAC-Seq and RNA-Seq to identify disease associated aberrantly regulated genomic region. We then use existing tools and develop novel genome and epigenome profiling and manipulation tools to study epigenetic mechanisms of chromatin structure and function.
• Assoc. Prof. Mehmet Remzi DOKMECI, PhD
Associated Professor at University of California at Los Angeles, CA, USA
Mehmet R. Dokmeci is currently an Adjunct Associate Professor in the Department of Radiology and a co-Director of the Center for Minimally Invasive therapeutics at UCLA. Previously he was an Instructor at Brigham and Women’s Hospital and Harvard Medical School. Trained an electrical engineer, his research is based on the development of novel technologies for the creation of micro and nanodevices for applications in tissue engineering and regenerative medicine. He has a strong background and more than a decade of experience in designing and fabrication of micro and nanoscale devices, microfluidics for applications in biomedical devices, tissue engineering and implantable biosensors. He has been actively involved in research areas such as flexible electronics for monitoring and modulation of wound healing, electrical/electrochemical biosensors and microfluidic systems for organs-on-a-chip applications, as well as 3D bioprinting. . He has extensively published in refereed journals and conferences in these areas and is the author of 116 technical journal articles and 110 conference publications.
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• Assist. Prof. Athanasia PAVLOPOULOU, PhD
Assistant Professor at the Izmir Biomedicine and Genome Institute, DokuzEylül University (DEU), Izmir, Turkey
Athanasia Pavlopouloureceived her BSc degree in Biology from Western Illinois University (WIU), USA, and her MSc in Molecular Biology from the University of Nebraska-Lincoln (UNL), USA. As a PhD student in the University of Patras, Greece, she developed and applied Bioinformatics tools for the phylogenetic analysis and prediction of protein structure/function. She was also trained in the University of Manchester, UK, in the construction of protein fingerprints. She has worked as a postdoctoral researcher in diverse projects. Her main research interests include application of bioinformatics methods in the analysis of biological sequences, comparative genomics, phylogenetics, next-generation sequencing data analysis, processing of biological networks, and meta-analysis of cancer genetics and genomics data.
• Dr. Atakan PEKER, PhD
Washington State University, Pullman, Washington, USA
Atakan Peker is a graduate of Bogazici University (Mech. Eng. & Physics) and received his Ph.D. in Materials Science from CALTECH. Dr. Peker is co-inventor of Liquidmetal series bulk amorphous alloys and has been a technical leader in its commercial development. His career spans leading roles from start- up to academic research with extensive experience on R&D, product development, intellectual property and technology transfer. He is inventor in more than 40 patents.
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• Dr. Semir BEYAZ, PhD
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA
Arriving at CSHL in December 2017, Beyaz has been received three different honors for his research, which looks at how dietary choices impact the body’s immunity against cancer. His research group investigates the causal cellular and molecular mechanisms that link nutrition to organismal health and disease. For example, diets that lead to obesity, such as high fat diets are significant environmental risk factors that influence cancer incidence and progression in several tissues. Although the interactions between tumor cells and the immune system play a significant role in tumorigenesis, little is known about how dietary perturbations impact immunity against cancer. Our studies interrogate the functional consequences of diets on immune recognition and response pathways that play critical role in cancer immunity. By identifying the altered gene expression and metabolic programs in the immune system in response to dietary perturbations, our goal is to uncover mechanistic links that can be therapeutically exploited for the treatment of diseases associated with immune dysfunction such as cancer.
• Selin ARSLANHANMEMİŞ
Founder at ReDis Innovation
Selin Arslanhan Memis is the founder and director of ReDis Innovation, an innovation interface based in Turkey. She specialized in technology and innovation ecosystem. Selin carries out projects on innovation and technology to contribute strategy and policy design for both public and private sectors. ReDis Innovation brings together corporates, startups, researchers and government institutions by establishing joint platforms and developing partnership programs between different actors in the ecosystem. Previously, she was the Director at TEPAV and established and ran the Innovation Studies Program there. Selin managed over thirty projects and programs such as "Public-Private Partnership Models in High- Tech Industries", "Startup Accelerator Programs", "Design of the Roadmap at the Pharmaceutical R&D Ecosystem".
Selin is also a columnist at Dünya Gazetesi. She holds BA in Molecular Biology and Genetics and also an MBA degree with a thesis on economic valuation of a biotechnology R&D project.
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Contents
Programme...... (16-19)
Oral presentations………………………………………………………(20-94)
Poster Presentations……………………………………………………...(95-114)
Author index………………………………………………………………..(115-118)
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Oral Presentation Abstracts for the SANKO University Innovation in Medicine Summit-IV 11th-13th Oct 2018, Gaziantep, Turkey
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S.1. ASSESSMENT OF IMMUNOTOXICITY AND CELLULAR UPTAKE OF CARBON DOTS FROM SUGAR BEET MOLASSES IN RAW 264.7 MACROPHAGES
Emine Yavuz1, Saliha Dinç 1,2, Meryem Kara 1,3
1 Advanced Technology Research and Application Center Selçuk University, Alaeddin Keykubat Campus Konya, Turkey. 2 Cumra School of Applied Sciences, Selcuk University, Konya, Turkey. 3 Selcuk University Cumra Vocational High School, Konya, Turkey.
Abstract—Foodborne carbon dots (CDs)- extracted with negative zeta potential (-5,37 mV) were uptaken from sugar beet molasses were firstly characterized efficiently into macrophages and accumulated in the by UV-Vis, photoluminescence, Fourier transform cytoplasm. CDs showed no cytotoxicity against infrared spectroscopy, dynamic light scattering, and mouse macrophages at concentrations 2.15 g/L and transmission electron microscopy. Elemental less, but LPS-induced NO production was determination of carbon dots was done by SEM- significantly decreased by CDs at non-toxic doses (p EDX. In order to investigate the effect of CDs on < 0.0001). Since NO is believed to be a major RAW 264.7 macrophage cells, immunotoxicity proinflammatory mediator, here in vitro studies have experiments were done by two different techniques: shown that CDs might have anti-inflammatory Colorimetric XTT cell viability assay and effects against the pathologic and excessive xCELLigence RTCA DP (real-time cell analysis production of NO. dual purpose) system. The dose-dependent effects of Keywords—Sugar beet molasses, Carbon dots, CDs on LPS induced NO production of macrophages Macrophages was investigated by Griess method. In order to determine the intracellular localization of fluorescent CDs confocal microscope was used. Fluorescence spectra exhibit the emission maximum (λem) at 465 nm when using an excitation wavelength (λex) of 370 nm for molasses C-dots. Monodisperse CDs 22 The Congress Is Supported By Turkish Airlines
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Introduction CDs formed naturally during refining process of
Biocompatible carbon dots are good alternative to sugar beet was also tested by confocal microscopy conventional semi-conductor quantum dots which imaging. have toxicity and environmental drawbacks [1]. Due According to the results of experiments CDs were to these advantages as well as the abundant and found biocompatible; decreased LPS induced NO inexpensive nature, carbon dots have found many production; were uptaken efficiently and applications in the fields of bio-sensing, drug accumulated in the cytoplasm of RAW 264.7 delivery, optoelectronics, photocatalysis, bio- macrophages. Immunotoxicity and determination of imaging [2-4]. Carbon dots (CDs) can be synthesized interlocalization of CDs may help to extend potential with different approaches such as electrochemical utilization of these biocompatible materials in and hydrothermal synthesis, combustion, medical applications. microwave, chemical oxidation, and more. Besides Experimental that, carbon dots can be easily obtained from commercially available different food sources [5-6]. Preparation and characterization of Carbon dots- Although these small nanoparticles are generally not 5 g of sugar beet molasses was dissolved in 10 mL of assumed as toxic material, evaluation of their deionized water and mixed to obtain homogenous immunocompatibility has a significant importance solution. Centrifugation was applied for 5 min at with regards to their effects on human health [7]. 5000 rpm and CDs containing supernatant was diluted with water in ratio of 1:80. UV–Vis In this study, endogenous carbon dots simply absorption of CDs was examined by UV-Vis extracted from sugar beet molasses, dark byproduct spectrophotometer (Biochrom Libra S22) and of refining of sugar beet factories, were tested for the fluorescent spectra were taken by first time to explore their effect on RAW 264.7 photoluminescence spectrophotometer (F-7000, murine macrophage cells. The effects of CDs on HITACHI, Japan). FTIR spectra were performed macrophage cell viability and metabolical activity using FTIR-ATR Bruker model (Vertex70 series) at were analyzed and non-cytotoxic doses were a resolution of 4 cm–1 to understand chemical groups determined. The dose dependent effect of CDs on on the surface of as prepared CDs. Elemental LPS induced NO production of RAW 264.7 cell line determination of carbon dots was done by scanning was investigated. In vitro cellular uptake of these 23 The Congress Is Supported By Turkish Airlines
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electron microscopy with Energy Dispersive X-ray reagent. xCELLigence RTCA DP analyzer (Acea Spectroscopy (SEM-EDX). EDX analysis was Biosciences, USA) was used for real-time dynamic performed using EDAX Genesis (123 ev) at analysis of cell viability of macrophages treated with accelerating voltage of 20 kV and a working distance different doses of carbon dots. The dose-dependent of 10 mm. Images of CDs were taken with TEM effects of CDs on proliferation of macrophages were (Transmission Electron Microscope, JEOL JEM- monitored for about five days (data acquired every 2100 UHR). For TEM measurements CDs were 15 minutes). To analyze the dose-dependent effects diluted with ethanol and dropped onto carbon coated of CDs on LPS induced NO production, RAW 264.7 copper grid. To determine surface charge of carbon cells were treated with sugar beet molasses sourced dots zeta potential studies were performed on a carbon dots for 24 h. To measure the level of NO, Zetasizer Nano-ZS model (Malvern Instruments Pvt. Griess assay that has been extensively used for the Ltd., UK). detection of NO through determining nitrite/nitrate concentration was applied. To investigate the In vitro cell experiments- BALB/c mice originated intracellular localization of CDs in macrophages murine macrophage cell line RAW 264.7 (TIB71) RAW 264.7 cells were seeded in special glass bottom was obtained from the American Type Culture 24-well plates (Sensoplate, Greiner Bione, USA) and Collection (ATCC, USA) and maintained in incubated for 24 h at 37 °C. Then, cells were treated supplemented Dulbecco’s Modified Eagle’s medium with different doses of CDs for 24 h, then membrane (DMEM) in 5% CO2, 95% air and humidified of the cells were labeled with Alexa Fluor 680 atmosphere at 37 °C. The relative cell viability was phalloidin which is a high-affinity F-actin probe. The determined by two different techniques: i) images were captured through a laser scanning colorimetric tetrazolium assay XTT, and ii) label- confocal microscope (Nikon A1R1, Japan). free monitoring of CDs-treated macrophages by xCELLigence RTCA DP system. Cells were seeded Results in 96-well plates at a density of 10,000 cells per well. A. Characterization of Carbon dots On the following day, cells were incubated with Figures 1, A and B show the UV-vis absorption and increasing concentrations of C-dot solution (up to 4.0 emission spectra for CDs, respectively. The emission mg·mL−1) and cytotoxicity was evaluated 24 h after spectra of carbon dots at different wavelengths using XTT (Biological Industries, Israel) viability
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showed excitation dependent behavior. The highest carbon dots, and used XTT cell viability assay to test fluorescence emission at 465 nm was recorded at 370 metabolic activity and xCELLigence RTCA system nm excitation wavelength. The size of to continuously monitor cell growth/inhibition. The monodispersed the carbon dots is around 2 nm xCELLigence RTCA analyzer measures electrical according to TEM image (data now shown). impendence across wells in which the attachment and proliferation of cells increases impendence (i.e. “cell index”).
When CDs were administered to RAW 264.7 cells (Fig. 2) all groups continued to proliferate as compared to non-treated cell control in a dose- dependent and time-dependent manner. IC50 values of carbon dots for 48h, and 72h were 400 µg/ml and 100µg/ml, respectively. For the first 24h of the treatment with carbon dots (except 4.3 g/L) proliferative and metabolically active phase of macrophages compared to non-treated cells was recorded (Fig.3).
Fig. 1. (A) UV-vis-spectrum of UV-Vis spectra of carbon dots (CDs). a: CDs under sunlight; b: CDs under 365 nm UV light. (B) Excitation-dependent fluorescence emission of CDs.
B. Cell viability measurements
To investigate the effects of CDs on cell Fig. 2. Continuous monitoring of sugar beet proliferation, we treated RAW 264.7 cells with molasses-treated macrophage cells. RAW 264.7 cells 25 The Congress Is Supported By Turkish Airlines
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were plated at 40000 cells/well of a 16-well E-plate and treatments were added at ~24 hours (arrow).
Fig. 4. Effect of CDs on nitric oxide production of RAW 264.7 macrophages for 24h (Griess Assay). Fig. 3. Effects of CDs on metabolic activities of RAW 264.7 cells for 24h D. Internalization of C-dot in RAW 264.7 cells
C. NO production As this study was interested in determining macrophage function after CDs accumulation, Following the stimulation of RAW 264.7 studies were conducted to verify CD incorporation macrophages with LPS (1 µg/ ml) and two different into the cells. Figure 5 shows internalization of the concentrations of CDs (2.15 g/L and 1.075 g/L) carbon dots into the cells via confocal microscopy. accumulation of nitrite in the cell supernatants was CDs were localized inside the cytoplasm. measured by a colorimetric assay based on the Griess reaction. While LPS alone markedly induced NO production, CDs significantly reduced the level of NO production in LPS-induced RAW264.7 cells (Fig. 4).
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References
[1] Pengli Zuo, Xiuhua Lu, Zhigang Sun, Yuhan Guo. Hua He..A review on syntheses, properties, characterization and bioanalytical applications of fluorescent carbon dots. 2015. Microchim Acta. DOI 10.1007/s00604-015-1705-3.
[2] Bibekananda De and Niranjan Karak. , 2016, Recent progress on carbon dot-metal based nanohybrids for photochemical and electro- chemical applications. Journal of Materials. Fig. 5. Confocal microscopy images of RAW 264.7 Chemistry ADOI: 10.1039/C6TA10220D cells following 24 h of incubation with CDs (2.15 g/L). Red is Alexa Fluor 680 phalloidin -stained cell membrane. Black dots within the cells are CDs [3] Mhetaer Tuerhong, XU Yang, YIN Xue-Bo. agglomerates. The scale bars correspond to 25 µm. 2017. Review on Carbon Dots and Their Applications. Chinese Journal of Analytical Conclusions Chemistry. 45(1), 139–150. Here we presented a detailed analysis of the effect of
CDs on macrophage function in vitro. RAW 264.7 [4] Kim Lategan, Hend Alghadi, Mohamed cells were exposed to different doses of CDs and Bayati, Maria Fidalgo de Cortalezzi, their viability, proliferation, NO production function, and Edmund Pool. Effects of Graphene Oxide and phagocytosis were evaluated. Results indicate Nanoparticles on the Immune System that sugar beet molasses-sourced C-dots have shown Biomarkers Produced by RAW 264.7 and immunomodulatory properties. It may be possible Human Whole Blood Cell Cultures. 2018. that CDs may be effective to prevent acute Nanomaterials (Basel), 8(2): 125. inflamation.
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[5] Dinc, S., Kara, M., Demirel Kars, M., Aykül, F., Çiçekci, H., Akkuş, M. (2017). Biocompatible yogurt carbon dots: evaluation of utilization for medical applications. Applied Physics A, 123 (572) (DOI 10.1007/s00339-017-1184-y).
[6] Dongmei Li, Xiaokang Na, Haitao Wang, Yisha Xie, Shuang Cong, Yukun Song, Xianbing Xu, Bei-Wei Zhu, and Mingqian Tan. Fluorescent Carbon Dots Derived from Maillard Reaction Products: Their Properties, Biodistribution, Cytotoxicity, and Antioxidant Activity. J. Agric. Food Chem. 2018, 66, 1569−1575.
[7] Zhongcai Gao, Guangxia Shen, Xiunan Zhao, Na Dong, Peiyuan Jia, Junhua Wu, Daxiang Cui, Yingge Zhang and Yuxia Wang.Carbon dots: a safe nanoscale substance for the immunologic system of mice. Nanoscale Research Letters 2013, 8:276
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S.2. DETERMINING PARTICLE NUMBER CHANGES BY USING BIOIMPEDANCE NEEDLE PROBE
Tuba Denkçeken
SANKO University, Faculty of Medicine, Department of Biophysics, Gaziantep, Turkey.
Objectives: To determine the numerical particle number in tissue phantom was tested variation of polystyrenes with a diameter of 4 µm during the study. Spectroscopic data were using a bioimpedance needle probe. evaluated with impedance values at 5, 50, 100 Methods: Bioimpedance needle probe was and 200 kHz and the information about the connected to a bioimpedance measurement particle number in the tissue phantom was device to record spectra in the kHz range. The obtained. Bioimpedance spectra results which 18Gauge 1.2x89 mm spinal needle was utilized were obtained from polystyrene microspheres to design probe which was used for obtaining showed that the relationship between the impedance information about the 4 µm diameter impedance values and the number of the particle polystyrene microsphere. The current was is directly proportional. transferred to the phantom and conductivity Conclusions: Due to the highly accurate information was obtained by the probe. differentiation of particle number in the tissue Results: Bioimpedance probe needle allows phantom, our probe has potential to be used in the discrimination of particle number based on their real-time detection of microorganism colony specific signature. The sensitivity of the probe to number without the need for culture.
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S.3. DETERMINATION OF GLUTATHIONE BY AN ELECTROCHEMICAL SENSOR BASED ON POLY-DOPAMINE
Ebru Kuyumcu Savan
Faculty of Pharmacy, İnönü University, Malatya, Turkey.
Objectives: It is important to detect Glutathione modified sensor using differential pulse and cylic (GSH) using a reliable method with good voltammetry techniques. Determination of GSH sensitivity and selectivity. Due to the in real samples was realized at proposed electrochemically oxidizable characteristic, there modified sensor. has long been a great deal of developments in Conclusions: Surface modification of the electrochemical determination of GSH. The aim electrode with PDA thin film allows analyzing of this study is to determine GSH using the alteration of the polymer electrochemical electrochemical sensors based on poly-dopamine response. Differential pulse voltammetry is (PDA). applied successfully for the determination of Methods: In this work, PDA thin film was trace amounts of GSH in the urine samples on constructed on glassy carbon electrode surface voltammetric response of the modified sensor. by electropolymerization method. PDA was The results suggested that the PDA modification synthesized using a facile electro-deposition by of electrodes may provide a new strategy for cyclic voltammetry technique. Differential pulse GSH concentration determination in and cylic voltammetry were used for quantitative physiological solutions. determination of GSH. References Results: The PDA thin film-based sensing 1. Yang, C. et al. New J. Chem. 41, 3374- platform exhibited high selectivity and 3379 (2017). sensitivity with low detection limit. 2. Heng, S., Zhang, X., Pei, J. & Abell, A. Electrochemical determination of GSH was D. Biosensors 7, (2017). carried out by electrochemically at PDA
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S.4. SIMULTANEOUS DETERMINATION OF SEROTONIN AND MELATONIN ON POLYIMIDE MODIFIED SCREEN-PRINTED CARBON ELECTRODE
Ensar EREL1, İmren ÖZCAN1, Süleyman KÖYTEPE1, Burhan ATEŞ1, İsmet YILMAZ1
1 Inonu University, Department of Chemistry, Malatya, TURKEY.
Melatonin and serotonin (5-HT) involves many In this study, polyimide films were synthesized biological and physiological processes of the from 2.7-Diaminofluorene, promellitic body. The main task of protecting the body's dianhydride and used to modification carbon biological clock, which set the rhythm of Screen-Printed electrode to elimination of the melatonin, with its lipophilic property, is a interferant species for simultaneous powerful antioxidant because it can reach all determination of melatonin and serotonin. body areas and easily cross the blood-brain Structural characterizations of selective films barrier. Serotonin is a monoamine were characterized by FTIR, DSC, DTA and neurotransmitters and is widely distributed in the TGA. Furthermore, melatonin and serotonin biological systems. 5-HT is a key mediator in the selectivity properties of polyimide film based depression symptoms and gastrointestinal electrodes was examined by differential pulse disorders. 5-HT in the blood may be regarded as voltammetry technique. The voltametric results biochemical markers for depression. Therefore, indicate that the polyimide film based electrodes fast determination of 5-HT in the blood is can be used as a sensor for simultaneous considerable to the diagnosis, illness monitoring determination of melatonin and serotonin with and result of the depressed patients. Melatonin the good sensitivity, selectivity, and rapidly and usually appears in the presence of 5-HT; high R-values (0,9805-0,9809). therefore, their simultaneous determination may be extremely important to understand biological systems and samples.
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S.5. PREPARATION OF SERICIN BASED UV-CURING MULTIFUNCTIONAL POLYURETHANE TISSUE ADHESIVES
Sevgi Balcıoğlu, Ahmet Ulu, Samir Abbas Ali Noma, Canbolat Gürses, Merve Gökşin Karaaslan, Süleyman Köytepe, Burhan Ateş
Department of Chemistry, Faculty of Science, Inonu University, Malatya, Turkey.
Each year, millions of people may be exposed to Methylenebis(cyclohexylisocyanate)/isophorone traumatic/surgical wounds, requiring a proper diisocyanate were used. The closure. Conventional closure techniques, such isocyanate/PEG/sericin/β-cyclodextrin ratio was as suturing and stapling, have many 100/105/5/10, respectively. For the second step, disadvantages that they lead to scarring in the 2-isocyanoethyl methacrylate was reacted with operation site, cause inflammation and require a free hydroxyl groups of polyurethanes followed longer healing process. These techniques are by exposure to UV-light for 15 min to obtain the often inadequate for severe wounds, in particular final product. According to the results, it was for the closure of liquids. Therefore, a wide seen that the adhesives were obtained variety of tissue adhesives, sealants and successfully according to the structural, hemostatic agents have been developed1. morphological and thermal analyses. However, studies are still ongoing since tissue Furthermore, adhesion strengths, adhesives with sufficient properties are not biodegradability, biocompatibility, antibacterial available. In order to obtain tissue adhesives with property, gentamycin release and protein sufficient properties, we synthesized sericin- adsorption studies showed that the adhesives had based UV-curable polyurethane-acrylates. The a sufficient level of adhesion (2000-4000 kPa) reaction was performed in two steps. For the and biological properties(15-30% weight loss; synthesis of the polyurethanes in the first step, as 85-98% cell viability; 40-170 µg protein/0.1g hydroxyl source; sericin, β-cyclodextrin and polymer adsorption). PEG200/PEG400 and as isocyanate source; 4,4′-
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As a summary, synthesized adhesives may be References;(1)Bouten,P.J.M.et.al.Prog.Polym.S used in the medical field thanks to sufficient ci.2014,39(7),1375-1405. mechanical and biological properties. This study was supported by grants from TUBITAK(KBAG-116Z501).
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S.6. CU2+-SELECTIVE AND REVERSIBLE CHROMOGENIC AND FLUOROGENIC SENSOR BASED ON A RHODAMINE DERIVATIVE
Mecit Ozdemir
Department of Food Processing, Vocational School of Technical Sciences, Kilis 7 Aralik University, Kilis, Turkey.
Copper is the third most abundant heavy environmental science, which offer a number of transition metal ion, it is an essential important advantages such as operational micronutrient for most aerobic organisms and simplicity, low cost, real-time analysis, fast plants. Copper also performs a vital role in response, high reliability, and selectivity and maintaining a healthy metabolism due to its sensitivity for HTMs in aqueous solutions6-10. redox activity as a cofactor in many In this work, a new colorimetric and fluorescent metalloenzymes. Many genetic disorders, ‘Off-On’ sensor RhCHN, prepared from including Alzheimer’s, Parkinson’s, Menkes, Rhodamine B hydrazide and dibenzalacetone, Prion, Huntington’s and Wilson’s diseases have was designed and synthesized for the selective been recently associated with the excessively detection of copper ions in aqueous media. Its accumulated copper, as a redox-active nutrient, structural confirmation was carried out by using in the human body from copper-contaminated FT-IR, 1H NMR, 13C NMR spectroscopies and water. On the other hand, copper deficiency Tandem MS spectrometry. In the presence of within the body can cause the risk of coronary Cu2+ ions, probe RhCHN showed the maximum heart disorders1-5. Unlike the traditional enhancements in both absorption and analytical methods (ICP-OES, AAS, AFS, ICP- fluorescence spectra centered at 565 and 588 nm MS, etc), colorimetric and fluorescent receptors in EtOH-H2O (2:1, v/v, 10 mM HEPES, pH 7.2), capable of binding to toxic transition metal ions respectively, accompanied by a drastic color have recently become the focus of interest of change from colorless to purple. Interestingly, scientists in the fields of chemistry, biology, and however, the addition of excessive amounts of
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the other interfering metal ions did not lead to 4) Kempahanumakkagaari, S. K., Thippeswamy, any changes in the solution color, UV-vis R., Malingappa, P., J Lumin. 2014, 146, 11-17. absorption and fluorescence spectra. In the 5) Jiang, Z., Tian, S., Wei, C., Ni, T., Li, Y., Dai, meantime, the binding interaction between L., Zhang, D., Sens Actuators B Chem.2013, RhCHN and Cu2+ ions undergoes through the 184, 106-112. chelation-induced the spirolactam ring-opening 6) Ding, H.; Li, B.; Pu, S.; Liu, G.; Jia, D.; Zhou, mechanism. Further, it was also determined that Y. Sens Actuators B Chem. 2017, 247, 26-35. chemosensor RhCHN is bound to Cu2+ ion in a 7) Udhayakumari, D.; Naha, S.; Velmathi, S. 1:1 coordination stoichiometry with a very low Anal. Methods. 2017, 9(4), 552-578. detection limit for Cu2+ (34 nM). On the basis of 8) Zhou, Y.; Wang, F.; Kim, Y.; Kim, S.J.; Yoon, the experimental results, it was concluded that J. Org. let. 2009, 11(19), 4442-4445. the molecular sensing platform RhCHN has excellent selectivity and sensitivity toward Cu2+ 9) Min, M.; Wang, X.; Chen, Y.; Wang, L.; Huang, H.; Shi, J. Sens Actuators B Chem. 2013, only with the reversible dual chromogenic and 188, 360-366. fluorogenic response over other important competitive metal ions. 10) Yu, C.; Wang, T.; Xu, K.; Zhao, J.; Li, M.; Weng, S.; Zhang, J. Dyes Pigm. 2013, 96, 38- 44. References:
1) Dong, M., Ma, T. H., Zhang, A. J., Dong, Y. M., Wang, Y. W., Peng, Y., Dyes pigm. 2010, 87(2), 164-172.
2) Wu, S. P., Huang, Z. M., Liu, S. R., Chung, P. K., J fluoresce. 2012, 22(1), 253-259.
3) Mergu, N., Gupta, V. K., Sens Actuators B Chem. 2015, 210, 408-417.
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S.7. ASSESSING TOXICITY OF NANOPARTICLES, COMMONLY USED IN
NANOMEDICINE, TiO2 AND Fe3O4, USING IN VITRO MODELS
Yuksel CETIN1, Yasemin YILDIZHAN1, Zelal ADIGUZEL1, Gamze GUNGOR1, Ezgi KIYGA1, Fatma Gozde YUCE2, Hatice DURAN2
1 TUBITAK MAM, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli/TURKEY. 2 TOBB University of Economics and Technology, Department of Material Science and Nanotechnology Engineering, Cankaya, Ankara/TURKEY.
Nanotherapeutics provides targeted drug HCT-116), human cercix adenocarcinoma delivery, improve drug solubility, extend drug (HeLa), human breast adenocarcinoma (MCF-7), half-life, improve a drug’s therapeutic index, in and non-cancer cell lines; human peripheral vivo imaging, in vitro diagnostics, biomaterials, blood monocyte (THP-1), Chinese hamster ovary and active implants. Although nanomedicine (CHO-K1). The genotoxic potential of NPs was provides important new tools to deal with the analyzed by performing in vitro comet assay grand challenge of an ageing population, they using A549 and Caco-2 cell lines. The effect of have to be strictly regulated and follow thorough NPs on oxidative stress was evaluated by characterization, and safety assessment. The analyzing cellular reactive oxygen species using main objective of this study is to evaluate toxicity 2’,7’–dichlorofluorescin diacetate (DCFDA). of nanoparticles (NPs), TiO2, Fe3O4 and (3- Even though the cytotoxic effects of tested NPs aminopropyl)triethoxysilane (APTES) coated were not too high, there are still concerns on Fe2O3 commonly used in nanomedicine using in possible cause to oxidative stress and vitro models. The cytotoxic effects of tested NPs genotoxicity which need to be confirmed with at the concentrations of 0.05, 0.1, 0.2, 0.4, 0.6, further in vivo studies to evaluate their safety. 0.8, 1.0 mM was investigated by using cancer Keywords: Titanium dioxide, Iron oxide cell lines from different origins; human lung nanoparticles, nanosafety assessment, adenocarcinoma (A549 and Calu-3), human nanomedicine, nanotoxicity, cytotoxicity, colon colorectal adenocarcinoma (Caco-2 and oxidative stress, genotoxicity, in vitro models 36 The Congress Is Supported By Turkish Airlines
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S.8. MAGNETIC FIELD ENHANCES CATALYTIC ACTIVITY OF L-ASPARAGINASE
CONJUGATED ON FE3O4-CHITOSAN NANOPARTICLES
Burhan Ates1, Ahmet Ulu1, Suleyman Köytepe1, Samir Abbas Ali Noma1, Veli Serkan Kolat2, Tekin Izgi2
1 Department of Chemistry, Faculty of Science & Arts, Inonu University, Malatya, 44280, Turkey. 2 Department of Physics, Faculty of Science & Arts, Inonu University, Malatya, 44280, Turkey.
Objectives, L-asparaginase (L-ASNase) is a wider range of pH stability in comparison with unique enzyme for the enzymatic hydrolysis of free L-ASNase. Moreover, the reusability of L-Asn in medicine and food industry (1). conjugated L-ASNase was significantly Nevertheless, the instability, toxic effect and improved after immobilization and it retained poor reusability of its significantly limit the long- 60.5% of its initial activity after undergoing 16 term applications (1). To overcome these cycles. Furthermore, we revealed that the activity disadvantages, magnetic carrier consisting of of conjugated L-ASNase onto magnetic Fe3O4- magnetic Fe3O4-chitosan nanoparticles are chitosan particles increased about 3-fold in the immobilized with L-ASNase. We also weak magnetic field at certain frequencies and hypothesized that magnetic excitation of flux density compared to free one. conjugated L-ASNase can improved catalytic Conclusions: As a result, the application of a activity of L-ASNase. static magnetic field can be a promising strategy Methods: L-ASNase was covalently for the increase of enzyme activity, especially immobilized onto Fe3O4-chitosan carriers to industrially important enzymes. This study was determine the effect studies of magnetic field on partially supported by from Inonu University conjugated L-ASNase activity. (FDK-2017-751). Results: The conjugated L-ASNase displays References enhanced catalytic activity in a weak magnetic 1. A. Ulu and B. Ates, Bioconjug. Chem., 2017, field, thermal and pH stabilities. The conjugated 28, 1598-1610. L-ASNase presented higher thermostability, a
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S.9. DEVELOPMENT AND CHARACTERIZATION OF ULTRASOUND SENSITIVE PACLITAXEL, SURVIVIN siRNA AND YM155 LOADED NANOBUBBLES AGAINST LUNG CANCER
Yücel Başpınar1, Gülşah Erel2, Mustafa Kotmakçı1, Hasan Akbaba1
1 Faculty of Pharmacy, Department of Pharmaceutical Biotechnology, University of Ege, 35100, İzmir, Turkey. 2 Faculty of Pharmacy, Department of Pharmaceutical Biotechnology, University of Izmir Katip Çelebi, 35620, İzmir, Turkey.
Objectives: The worldwide cancer-related death revealed great improvements. One of the most rate for lung cancer is about 23 %, more than the investigated carriers for drug targeting in sum of breast, colon and prostate cancer. chemotherapy in the last years are NBs. Due to Including lung cancer, almost each cancer type the fact, that the particle size of NBs (1-500 nm) has a great deal in common, an overexpression of is bigger than the pore size of healthy tissues, it the apoptosis inhibitor survivin (S). One serious is not possible for the drug to leave the blood disadvantage of chemotherapy with anticancer stream and enter the healthy tissues. However, drugs is the fact that after the parenteral the pore size of cancer tissue is big enough for an application of these drugs their distribution into entrance of the carrier and the drug. After the whole body via the blood circulation, but only reaching the cancer tissue, the drug carrier can a small amount of the dose can reach the tumour. enter the tumour tissue through the pores, where Anticancer drugs are not only potent against the the drug can accumulate. Because of being not tumour, they also harm healthy cells like bone able to enter healthy results in a low probability marrow, digestive system, reproductive system of undesired side effects. The aim of this study is hair follicels, leading to serious, but not desired to develop ultrasound sensitive NBs containing side effects. A useful strategy to reduce these side paclitaxel (PTX), survivin-siRNA and survivin effects of chemotherapy is drug targeting by the inhibitor sepantronium bromide (YM155), use of nanobubbles (NBs). Thus, it is possible to respectively, and to characterize the particles. To use nanotechnolgy for drug targeting, which the best of our knowledge there is no report in the
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literature about NBs containing PTX, YM155 Results: The blank NB formulation after the and survivin-siRNA for the therapy of lung optimization process had a particle size of 178 cancer. nm, a PDI of 0.41 and a ZP of 38 mV. The Methods: For developing the NBs, the lecithin particle size (308 nm), PDI (0.44) and ZP (53 concentration (1, 2 and 3 %), the chitosan mV) of the NBs prepared with DDAB were concentration (0.25, 1 and 2.7 %) and its increased. After adding PTX to the NBs, the molecular weight (low molecular weight and particle size (335 nm) and the PDI (0.57) futher medium molecular weight), the effect of increased, while the ZP (34 mV) decreased. The filtration through a 0.4 µm filter (with and PTX and YM155 loaded NBs had compared to without filtration), the effect of the stabilizers PTX loaded NBs had decreased particle size (76 Pluronic F68 (with and without Pluronic F68) nm) and PDI (0.24), and increased ZP (50 mV). and didodecyldimethylammonium bromide a) (DDAB) were tested. The NBs were characterized in terms their particle size, size distribution (Polydispersity index, PDI), zeta potential (ZP), with dynamic light scattering using non-invasive back scattering technique and laser doppler micro-electrophoresis technique, respectively. Furthermore, the morphology and cytotoxic effects of the NBs were investigated by b) using confocal laser scanning microscopy (CLSM), and colorimetric XTT on the lung cancer cells A549. After these formulation parameter and the preparation method were optimized, the preparation temperature was investigated and PTX, YM155 and siRNA, respectively, were loaded into the NBs.
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Figure 1: Schematic illustration (a) and CLSM image (b) of blank nanobubble.
The CLSM image reported (Figure 1b) showed that the NBs own a spherical morphology with a well-defined core–shell structure. Similar shapes were observed for all the other NB formulations.
Even the application of low concentration (4 µL/well) of the PTX and YM155 loaded NBs resulted in significant decrease of the viability of A549 lung cancer cells. Conclusions: For preparing suitable NBs with a small particle size, a narrow size distribution and a high ZP, 2 % lecithin, 0.25 % medium molecular weight chitosan, filtration through a 0.4 µm filter, 0.01 % of the stabilizing agent Pluronic F 68 and DDAB were necessary. Furthermore, these NBs revealed an enhanced cytotoxic effect on A549 lung cancer cells. Acknowledgement: This study has been financially supported by TUBITAK under grant code 116S213.
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S.10. LAB-ON-A-CHIP FOR CANCER CELL BIOLOGY
Devrim Pesen Okvur
Izmir Institute of Technology, Department of Molecular Biology and Genetics, Urla, Izmir, Turkey. INITIO Biomedical Engineering Consulting Industry and Trade Limited Company, Izmir Teknopark, Urla, Izmir, Turkey.
Cancer cell biology is complex in both space and surface protein patterns, our results showed that time. Microfluidics based lab-on-a-chip (LOC) distribution of invadopodia were polarized devices provide control at the nano/micrometer towards non-adhesive areas. and seconds scales. We designed, fabricated and Furthermore, we investigated interactions of used LOC devices to investigate cell adhesion, breast cancer cells with macrophages, a invasion and migration, which are the cellular prominent duet of the tumor microenvironment. phenomena that are uncontrolled in cancer Our results showed that the EGF (epidermal metastasis. growth factor)– CSF-1 (colony stimulating factor With respect to cell adhesion, using basic LOC 1) loop is a paracrine – juxtacrine loop contrary devices and complex nanopatterned surfaces, we to the generally accepted double paracrine loop. showed that breast cancer cells better adapted to polarization inducing conditions such as interstitial flow and nano-patterned gradient surfaces, demonstrating their plasticity in comparison to normal mammary epithelial cells. In regard to invasion, it was not known whether the cellular distribution invadopodia, which are proteolytic structures formed by cancer cells, can be regulated by the organization of the extracellular matrix. Using nanometer scale
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Finally, we are developing a new LOC based method for early detection of cancer metastasis. Here, we focus on the extravasation stage of the process, which we mimic in a LOC device.
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S.11. LOW-COST MICROFLUIDICS FOR MONITORING MINIMAL RESIDUAL DISEASE (MRD)
Ünal AKAR1 , Kutay İÇÖZ 1,2
1 Abdullah Gul University, Life and Natural Sciences Faculty, Bioengineering Department, Kayseri, Turkey. 2 Abdullah Gul University, Engineering Faculty, Electrical and Electronics Engineering Department, Kayseri, Turkey.
Abstract laboratory environment with include valves, The existence of residual malignant cells that pumps, and sort of other control methods. cannot be detected by morphological In this work, we have investigated a low-cost, examination of the bone marrow, can cause microfluidic-based alternative method for relapses and repeat leukemia. Flow cytometry or monitoring (MRD). polymerase chain reaction techniques are Keywords: MRD, Microfluidics, Surface becoming routine to follow up patients to detect Functionalization, Cellphone Microscope. MRD and adjust the treatment intensity. Methods However, these techniques are expensive and The microfluidic system is composed of require trained personnel, thus patients in low- polymethylmethacrylate (PMMA) and glass income countries do not have access to regular layers. On the glass layer, a matrice of gold were MRD monitoring. functionalization with antibodies to capture A microfluidic technique that is a novel approach target cells, cellphone microscopy was used to for detecting and sorting cells depend on their record images and count the cells. properties such as physical, chemical and Functionalization method uses the combination biological. Microfluidic systems, also called lab- of 11-Mercaptoundecanoic acid (MUA) self- on-chip systems, depending on a variety of the assembled monolayer and N-Ethyl-N’-(3- target matter that can be proteins, cells, SS dimethylaminopropyl) carbodiimide (Single-stranded) DNAs, single-cell molecules hydrochloride(EDC)/Nhydroxysulfosuccinimide etc. In a microfluidic channel, we can mimic a
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(NHS) linking with protein G modification. The 2.3 Modification of Chip Surface orders of the immobilization process are: The chip surface was modified with 1mM MUA Gold MUA EDC=NHS Protein G in ethanol for 24 hours then the surface was CD19 BSA B lymphoblast cells rinsed with ethanol. EDC/NHS was used as a
2.1 Materials cross-linker in the experiments. For EDC/NHS crosslinking the solution of 5 mg EDC, 5 mg B lymphoblast cells CCRF-SB were purchased NHS in 1 mL deionized water was put on the from ATCC (Rockville, MD), protein G was surface for 30 minutes, and the surface was purchased from ThermoScientific (Pittsburgh, rinsed with deionized water for 2 minutes. PA). The CD19 antibody was purchased from Subsequently, the protein G solution, 800 mL BioLegend (SanDiego, CA) and all other protein G (5% protein G solution in deionized chemicals were purchased from Sigma-Aldrich water) in 5.4 mL PBS was put on the chip surface (St.Louis, MO). for 25 minutes, and then surface rinsed with PBS 2.2 Preparation of Chip Surface for 2 minutes. The 500 mL of stock solution of CD19 (20 mL CD19 antibody in 1 mL PBS) was The immobilization of B lymphoblast cells on a diluted in 5.4 mL PBS and then the solution of gold-coated glass surface is the most important CD19 was put on the surface for 25 minutes, part of the experiments. Before starting to make followed by PBS. The last step before adding a self-assembled monolayer, a new surface was cells was prepared 5% BSA solution (0.5 mg respectively rinsed with deionized water and BSA in 10 mL PBS) and put on the surface to absolute ethanol and then dried under a nitrogen prevent non-specific bindings of cells for 35 stream. The chip was next put in acidic piranha minutes. Eventually, B lymphoblast cells in PBS solution (1 ml deionized water, 1 ml sulfuric was put on the surface in a microfluidic channel. acid, 1 ml hydrogen peroxide) was diluted in 10 A peristaltic pump was used to make flow ml deionized water for 1 minute then rinsed with (10µl/min) for 60-90 minutes. deionized water and dried under a nitrogen stream.
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Results concentration in solution directly affects the number of captured cells and proper working of This surface functionalization method based on the experiments. In Figure 2, captured cells the thiol group which means a covalent bond observed by an optical microscope recorded between sulfur in MUA and gold surface. As a using Nikon, microscope equipped with a result of this covalent bound, the importance of charge-coupled (CCD) camera. the exposure time to MUA increasing. In Figure 1, different exposure times to MUA are given. The gold-coated glass surface exposure to MUA for 30-60 minutes during different experiments and the efficiency of captured cells around 45% of captured cells and chip surface ratio. However, we observed around 80% efficiency while surface exposure time to MUA was around 24 hours.
Fig.1. Different exposure time to MUA during experiments
Furthermore, another important issue is BSA Fig.2. Captured Cells on Gold-Coated Glass Surface recorded by optical microscope using blocking for preventing non-specific binding of Nikon with a charge-coupled (CCD) camera. cells. Exposure time to BSA or BSA
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Figure 3 shows a microfluidic system and Figure Conclusion 4 shows square gold pads (30µm x 30µm) on a We built a low-cost microfluidic system to glass substrate which recorded by cellphone capture B lymphoblast cells and used cellphone microscope. The red arrows specify the cells microscopy to record images and count cells. The captured on the surface. The experiments showed current cell capture efficiency is 75%. Depends that as the amount of exposure to MUA on the current control experiments, we have increased, the number of captured cells captured 5 times more cells than the control increased. surface and the cost of a single chip is almost 300 TL. Considering the situation of exposure time to MUA, we should try to optimize exposure time during experiments.
Acknowledgement Authors acknowledge The Scientific and Technological Research Council of Turkey (TÜBİTAK Project No: 115E020) for financial Fig.3. A microfluidic system includes gold- support. coated glass surfaces
Fig.4. Captured cells on gold-coated glass surface recorded by cellphone microscope 46 The Congress Is Supported By Turkish Airlines
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S.12. BREAST CANCER AND EXOSOMAL SURVIVIN
M. Yıldırım1, H. Çiçek2, G. Nacarkahya3, Özlem Nuray Sever4, Necla Benlier5, Zeliha Yıldırım6
1Department of Medical Oncology, Faculty of Medicine, Bahcesehir University, Istanbul, Turkey. 2Department of Medical Biochemistry, Faculty of Medicine, Gaziantep University, Gaziantep, Turkey. 3Department of Medical Biology and Genetics, Faculty of Medicine, Gaziantep University, Gaziantep, Turkey. 4Department of Medical Oncology, Faculty of Medicine, SANKO University, Gaziantep- Turkey. 5Department of Medical Pharmacology, Faculty of Medicine, SANKO University, Gaziantep- Turkey. 6Department of Veterinary, İslahiye Vocational High School, Gaziantep University, Gaziantep, Turkey.
Objectives: The aim of this study is to determine included in the study. Exosomal survivin levels the role of exosomes and exosomal survivin in were 2.41 ± 6.22 (Range 0-40.52) ng / ml in the relation to breast cancer which increasingly breast cancer patients and 0.23 ± 0.62 (Range 0- understood in carcinogenesis. 2.4) ng / ml in the control group. The difference Methods: In the Medicalpark Gaziantep between the two groups was investigated by Hospital Medical Oncology Clinic, between Mann Whitney U test. A statistically significant 2014 and 2017, the patients whose diagnosis was difference was found between the patients with confirmed histopathologically and who were breast cancer and the control group (p = 0.037). diagnosed as invasive ductal cancer of breast and Conclusions: It has been shown that survivin, healthy volunteers were taken to the study. which was previously shown as a poor prognostic Serum samples from patients and healthy marker in the literature in breast cancer patients, volunteers were stored at -80. Exosomes were is exosomally expressed. Exosomal survivin isolated from these serum samples. The integrity expression is higher in breast cancer patients than of the obtained exosomes has deteriorated. in control group. This pathway can be both a Survivin levels were measured by ELISA therapeutic goal and a new marker for early method. recognition of the disease. Results: A total of 77 participants, 57 with breast cancer and 20 with healthy volunteers were 47 The Congress Is Supported By Turkish Airlines
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S.13. INVESTIGATION THE EXPRESSION OF ADAMTS9 GENE IN LIVER OF HIGH-FAT DIET AND STREPTOZOTOCIN-INDUCED DIABETIC RATS
Lütfiye Özpak1, Ayfer Pazarbaşı1, Mehmet Bertan Yılmaz1, Işıl Öcal2
1 Medical Biology, Çukurova University, Faculty of Medicine, Adana, Turkey. 2 Biophysics Department Çukurova University, Faculty of Medicine, Adana, Turkey.
insulin tolerance test were carried out. All of rats were sacrificed at the end of 16 weeks and livers Objectives: There are many risk factors for type 2 diabetes (T2D). One of the single best indicator of Type 2 diabetes is obesity. Insulin resistance were removed for molecular analysis. Real Time plays a primary role in the hepatic excess at PCR was performed to compare the expression accumulation of fats. Peripheral insulin levels of ADAMTS9 gene in groups. resistance is give rise to hyperglycaemia as well Results: ADAMTS9 mRNA expression level of as promotes the synthesis of fatty acids in the HFD-STZ induced rats were lower than control liver. In this study, we evaluate expression levels group. of ADAMTS9 gene that encoded liver proteins in Conclusions: Consistent with previous studies, T2D model of rats induced by Streptozotosin which were determined as candidate genes in the (STZ) and High Fat Diet (HFD). pathophysiology of obesity and T2D Methods: A total of 40 Wistar male adult rats development, ADAMTS9 gene were found to were divided into 4 groups of control (n=10), play a possible role in the progression of these STZ (n=10),STZ-HFD (n=10) and HFD (n=10). diseases. Rats were fed with high-fat diet (involved %50 This study was supported by Cukurova fat) for 16 weeks received of STZ (40mg/ml) University Scientific Research Projects Unit single dose with intraperitoneal injection and (Adana, Turkey) with TSA20179090 number.
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S.14. MRI FINDINGS AND PREVALENCE OF SKELETAL COMPLICATIONS IN CHILDREN AFTER HEMATOPOIETIC STEM CELL TRANSPLANTATION
Taner Arpacı
Acibadem University, Acibadem Adana Hospital, Department of Radiology, Seyhan-Adana, Turkey.
Objectives: To evaluate magnetic resonance Avascular necrosis and bone infarct were the imaging (MRI) findings and prevalence of most common findings (9 patients, 38% each). skeletal complications after hematopoietic stem- Avascular necrosis most frequently occured in cell transplantation (HSCT) in children. hemoral head (4 patients, 44%). Bone infarct was most commonly detected in tibia (6 patients, Methods: From February 2013 to May 2018, 67%). Myositis emerged in 4 patients (17%) and 225 pediatric patients with hemato-oncological most commonly involved crural muscles (3 diseases underwent HSCT. Twenty-four (11%) patients, 75%). Joint effusion was seen in 4 patients (13 girl, 11 boy; age range, 3-19 years; patients (17%), medullary edema was observed mean age, 14 years) who presented with skeletal in 3 patients (13%) and bone fracture was present symptoms and underwent MRI after HSCT in 3 patients (13%). One patient (4%) included in study. Eight (33%) of 24 patients demonstrated bursitis (retrocalcaneal) and 1 were diagnosed with acute lymphoblastic patient (4%) showed synovitis (hip joint). leukemia (ALL), 7 (29%) were with thalassemia, Conclusions: Prevalence of skeletal 3 (13%) were with acute myeloid leukemia complications after HSCT is reported as 13-20% (AML), 2 patients (8%) with aplastic anemia, 1 in the literature which was found 11% in this patient (4%) with Fanconi anemia, sickle cell study. MRI is essential for diagnosis of skeletal anemia (SCA), hemophagocytic syndrome and symptoms since early diagnosis and treatment neuroblastoma. improve the life quality of children after HSCT. Results: Twenty-two (94%) of 24 patients demonstrated skeletal MRI abnormalities.
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S.15. DIAGNOSTIC ROLE OF SERUM LL-37 LEVELS IN DIFFERENTIAL DIAGNOSIS OF PROSTATITIS AND PROSTATE CARCINOMA
Necla Benlier1, Mehmet Solakhan2, Özlem Nuray Sever3, Zeliha Yıldırım4, Hülya Çiçek5
1Department of Medical Pharmacology, Faculty of Medicine, SANKO University, Gaziantep-Turkey. 2Medical Park Hospital Department of Urology, Gaziantep-Turkey. 3Department of Internal Medicine, Faculty of Medicine, SANKO University, Gaziantep-Turkey 4 Department of Veterinary, İslahiye Vocational High School, Gaziantep University, Gaziantep, Turkey. 5Department of Medical Biochemistry, Faculty of Medicine, Gaziantep University, Gaziantep, Turkey.
Objectives: In our study the diagnostic role of 144.1±262.1 (4-1200) ng/ml in patients with LL-37 levels measured in serum were prostate carcinoma and as 54.5±115.4 (3.9 - 405 investigated in prostatitis and prostate carcinoma ng/ml) in patients with prostatitis. Statistically diagnosis and in the differentiation of these two significant difference was detected (p=0.036). conditions. Serum LL-37 was detected as 3.17±1.17 (1.63-
Methods: In Gaziantep Medical Park Hospital 7.01) ng/ml in the prostatitis group, 3.76±1.8 Urology Clinic between 2014 and 2017, patients (1.32-8.23) ng/ml in prostate carcinoma patients diagnosed with prostate cancer and prostatitis and as 2.29±1.19 (0.8-4.89) ng/ml in the control who were diagnosed as histopathologically and group. Significant difference was detected healthy volunteers were included in the study. between the three groups (p=0.008). A Serum LL-37 levels were investigated using significant relation was not detected when LL-37 values were compared with prostate cancer risk ELISA method. patients with prostate cancer and prostatitis, and healthy volunteers were included groups (p=0.714). in the study. Conclusions: We showed that LL-37 levels
Results: A total 52 patients consisting of 24 measured in serum were significantly higher in diagnosed with prostate carcinoma and 28 with patients diagnosed with prostate carcinoma. prostatitis, together with 18 healthy subjects were Measuring LL-37 in serum could be used as a included in the study. PSA was detected as diagnostic marker in prostate carcinoma as well 50 The Congress Is Supported By Turkish Airlines
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as could facilitate differential diagnosis to strengthen diagnostic suspicion before prostate biopsy and to distinguish from prostatitis cases.
Keywords: Prostate cancer, Prostatitis, LL-37
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S.16. EFFECT OF HISTAMINE ON REPRODUCTIVE HORMONES IN MALE RATS Burcin Altinbas1,3, Murat Yalcin1, Nasir Niaz1,2, Gokcen Guvenc1
1 Department of Physiology, Faculty of Veterinary Medicine, Uludag University, Bursa 16059, Turkey. 2 Department of Biosciences, University of Wah, Wah Cantt, Pakistan. 3 Department of Physiology, Faculty of Medicine, Sanko University, Gaziantep 27090, Turkey.
Objectives: Histamine is an effective hemocytometer was used, and for sperm neurotransmitter and neuromodulator in central morphology, smears were stained by Giemsa. nervous system. Histamine has been observed to Statistical analysis was performed using RM- mediate reproductive hormones in female rats, ANOVA and the post-ANOVA test of whereas there has been no concrete results on Bonferroni (P < 0.05) was considered significant. their male counterparts or the receptors involved Results: 100 nmol histamine only caused in the process. In this study, we investigated the significant increase in plasma samples. Besides, effects of the central histaminergic system on central pre-treatment with H1 and H2 receptors reproductive hormones (GnRH, LH, FSH, (100nmol) completely blocked the effect of testosterone) and sperm parameters, also histamine on hormones, whereas H3/H4 assessed the effects of the central histaminergic receptors (100nmol) had no effect. Neither receptors (H1, H2 H3/H4) on histamine-evoked histamine treatment alone nor its injection upon hormonal and sperm parameters. histamine receptor antagonist pre-treatment did Methods: In our study, 77 male adult Sprague- not alter any sperm parameters. Dawley rats were used. A total of 50 or 100 nmol Conclusion: Although centrally injected histamine were injected histamine caused increases in reproductive intracerebroventricularly. In order to measure the hormones, no effect on sperm parameters was hormone levels in the blood, the left femoral observed. Furthermore, these effects of histamine artery was cannulated, and samples were are mediated by central histaminergic H1 and H2 measured by ELISA. While semen collected by receptors. This study was supported by diffusion from epididymis, its motility examined Commission of Scientific Research Projects of by phase contrast microscopy. For analysis of Uludag University (OUAP(V)2015/26). sperm count, the chambers of Neubauer-type
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S.17. UP-REGULATED miR-125b-5p; A POTENTIAL CIRCULATING BIOMARKER IN POSTMENOPAUSAL OSTEOPOROSIS
Elif Pala
SANKO University, Faculty of Medicine, Department of Medical Biology, Gaziantep, Turkey.
Objectives: Osteoporosis is a disease for selection of eligible studies were defined as characterized by susceptibility to bone fragility follows: studies that report miRNA expression and increased risk of fracture. The most profiling on humans of Postmenopausal important parameter determining the fracture risk Osteoporosis (PMO); studies that compare PMO is bone mass and there is a strong inverse serum samples with healthy control serum relationship between Bone Mineral Density samples; studies that report significance of (BMD) and fracture risk. Currently, non-invasive p<0.05 in dysregulated miRNAs. Dual Energy X-ray Absorptiometry (DEXA) is Results: As a result of the literature search, 28 used as the gold standard in the evaluation of miRNAs were described to be dysregulated in BMD, but the disadvantage of this method is that serum, among them 15 were up-regulated, 9 were patients are exposed to radiation. MicroRNAs down-regulated and 4 showed both (miRNAs) are non-coding RNAs in 22 characteristics. Of all miRNAs, the mostly nucleotides length which are found in a wide determined miR-125b-5p has been shown to variety of organisms and play a role in many exhibit up-regulated expression pattern in 4 biological processes. miRNA expression studies different studies. allow early detection of various diseases like osteoporosis, gives opportunity to follow up the Conclusions: While the role of miR-125b-5p in progression and treatment response of the PMO needs to be investigated further, miR-125b- disease. 5p may be used as a potential noninvasive biomarker in this disease. Methods: Various databases were scanned using the appropriate keywords. The inclusion criteria
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S.18. EFFECTS OF SOME CHEMICALS ON THE ACTIVITY OF HUMAN ERYTHROCYTES GLUTATHIONE S-TRANSFERASE
Uğur GÜLLER1, Pınar GÜLLER2
1Igdir University, Faculty of Engineering, Department of Food Engineering, 76100-IĞDIR. 2Atatürk University, Science Faculty, Department of Chemistry, 25240-ERZURUM.
ABSTRACT While antipyrine and pyramidon did not show Glutathione S-transferases are a multifunctional inhibitory effect, sorbic acid and morin inhibited enzyme family that have an important role in GST with the IC50 value of 1.45 mM and 0.14 phase II of detoxification. GST catalyses mM respectively. The findings from this study to conjugation between electrophilic xenobiotic make people aware of the effects of these (such as drugs, toxins, environmental pollutants, chemicals which we are expose to in daily life on oxidative stress products and carcinogens) and GST activity. reduced glutathione. As a result of conjugation with GSH via GST, these compounds gain INTRODUCTION hydrophilic character and can be easily removed People are exposed to the effects of many foreign from the body. Considering these functions of chemicals (xenobiotics) due to drugs, food GST, inhibition of enzyme activity can be said to additives and environmental pollutants. Since cause significant problems for living organisms. xenobiotics enter metabolism, they are exposed In this study, GST enzyme was purified in a to various enzymes by detoxification reactions. single step by using Glutathione Agarose affinity These detoxification reactions are composed of chromatography from human erythrocytes. The Phase I, Phase II and Phase III reactions. Phase I inhibitory effects of sorbic acid, morin, reactions are reduction and hydrolysis reactions antipyrine and pyramidon were examined on and are catalysed by enzymes known as GST activity, obtained with 6.82 EU/mg protein cytochrome P450 (monooxygenase). In Phase II, specific activity. it is ensured that the compounds that are added
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the active group in phase I are converted into experimental and clinical trials have been made water-soluble metabolites. Glutathione S- (7). transferase (GST; EC 2.5.1.18) plays an MATERIALS AND METHODS important role in phase II reactions of Materials detoxification (1,2). GST enzymes catalyse the Sigma-Aldrich (St. Louis, MO) brand attack of sulphur atom of glutathione, a glutathione-agarose was obtained. 1-Chloro 2,4- tripeptide, to the electrophilic centre of various dinitro benzene (CDNB), reduced glutathione xenobiotic. As a result of this attack, the (GSH), and the protein assay reagent were electrophilic centres of these compounds are obtained from the Sigma Chem company (St. neutralized and their solubility in water Louis, MO). All the remaining chemicals were increases. These conjugates are inactivated by obtained from Fluka or Sigma (St. Louis, MO). discharging out of the cell with a glutathione Activity assay conjugate express (GS-X) pump (ATP- Enzymatic activity was determined dependent pathway) (3,4). GST also reduces spectrophotometrically with a Shimadzu cytotoxicity of some molecules such as bilirubin, spectrophotometer (UV-1208) at 280 nm and bile salts and drugs by binding them and 25°C according to the method described by increasing their mobility (5). Any damage that Habig et al. 1974 (8). may occur during the activities of the enzymes Protein Assay involved in the biotransformation blocks the Quantitative protein determination was measured detoxification and toxic compounds damage the spectrophotometrically at 595 nm according to organism (6). the Bradford method (9). The inhibition of these enzymes acting in cell Preparation of the hemolyzate defence is an important therapeutic step. For Erythrocyte pellet was obtained from Atatürk these reasons, it was thought that the use of GST University, Medicinal Faculty Blood Centre. inhibitors in chemotherapy might be beneficial. Erythrocyte pellet haemolysed with 5 volume of For this purpose, some compounds which are ice-cold water and then centrifuged at 10 000g effective on this system have been developed and for 30 min (at 4°C,) to remove the residual intact cells and membranes (10).
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Purification of glutathione S-transferase from reactions catalyse the conjugation of activated human erythrocyte xenobiotics with endogenous water-soluble The hemolyzate was applied to the glutathione- substances such as glutathione, UDP-gluconic agarose affinity column previously prepared acid or glycine (2). It has been reported that the equilibrated with 10mM pH 7.4 buffer GST enzyme plays a role in the development of (phosphate) with 150mM NaCl content. An multiple drug resistance in cancer cells (11). equilibrium buffer was used to wash the column Therefore, the enzyme has become a therapeutic and then the enzyme was eluted with a gradient target in the development of chemotherapy of GSH (5–10 mM) in a 50mM pH 9.5 Tris drugs. Multidrug resistance is the most important buffer. of resistance mechanisms developed by In Vitro Inhibition Studies malignant tumor cells against chemotherapy Inhibitory effects of sorbic acid, morin, drugs. Determining how various chemicals antipyrine and pyramidon were examined on interact with specific enzymes is very useful for enzyme activity. On this purpose, enzyme pharmacologists in terms of drug design. In the activities in varying concentrations of inhibitors treatment of diseases, targeting the necessary were measured and half maximal inhibitory enzymes in the respective metabolic pathway is a concentrations (IC50) were calculated by drawing very rational approach. “activity%-[I]” graphs. In this study the purification of the erythrocyte GST was performed by the single RESULT AND DISCUSSION chromatography step method by means of GST serves in phase II reactions in detoxification Glutathione-Agarose affinity gel of xenobiotics. In Phase I and Phase II reactions chromatography. The enzyme was purified of detoxification metabolism, a lipophilic and 1217.4-fold with a specific activity of 6.82 non-polar compound is transformed into a more EU/mg protein and overall yield of 25.78% water-soluble and less toxic metabolite which is (Table 1). more easily eliminated by the cells. Phase I reactions are mainly catalysed by the cytochrome
P450 system. Enzymes acting in phase II
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Table 1. Purification steps of glutathione reductase from human erythrocytes
Sample Type Total Total Total Specific Activity Protein Yield Purification Volume Protein Activity Activity (EU/ml) (mg/ml) % Coefficient (ml) (mg) (EU) (EU/mg) Hemolyzate 20 0.121 21.6 432 2.42 0.0056 100 1 Affinity 1.5 0.416 0.061 0.0915 0.624 6.82 25.78 1217.4 Chromatography
There are several studies on GST inhibtors in 6.25 µM, respectively (13). The inhibitory effects literature. Natural compounds including of sorbic acid, morin, antipyrine and pyramidon flavanoids derivatives such as baicalin, baicalein, on the activity of human erythrocyte GST were phloridzin, and phloretin have been reported to also examined in this paper. The inhibitor inhibit the human erythrocyte glutathione S- concentration which reduced the enzyme activity transferase with an IC50 value of 28.75, 57.50, by half was calculated from % activity-
769.10, and 99.02 µM, respectively. The Ki [inhibitor] graphs (Figure 1). It was observed that constants were calculated and exhibit Ki value of only sorbic acid and morin inhibited GST with
14.50 ± 0.71, 24.33 ± 2.08,762.50 ± 85.97, and the IC50 value of 1.45 mM and 0.14 mM 86.49 ± 1.11 µM, respectively (12). Other respectively. Lineweaver and Burk graphs reported inhibitors of GST enzymes were (1934) were drawn by using 1/V vs. 1/[S] values chalcone derivatives including 4- (Fig. 2), and Ki constant were calculated from Fluorochalcone, 4′-Hydroxychalcone, 2′- these graphs as 1.24±0.3 mM and 0.063±0.025 Hydroxy-4-methoxychalcone, 4,4′- mM for sorbic acid and morin that showed Diflurochalcone and 4-Methoxychalcone with an inhibitory effects on the enzyme. Inhibition types
IC50 value of 9.49, 9.76, 15.2, 24.75 and 25.66 were determined as non-competitive for each
µM respectively and Ki value of 7.76 ± 4.27, 12.8 inhibitor. ± 7.61, 15.36 ± 3.78 4, 19.5 ± 6.76 and 41.93 ±
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Figure 2. Lineweaver-Burk graph in 5 different Figure 1. Activity%-[inhibitor] graphs for (a) substrate (GSH) concentrations and in 3 different sorbic acid and (b) morin chemicals. inhibitor concentrations a) sorbic acid b) morin
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REFERENCES 8. Habig, W.H., Pabst, M.J., Jakoby, W.B., 1. Sinnett, D., Krajinovic, M., 2000. Leuk 1974. The Journal of Biological Lymphoma., 38(5-6), 447-62. Chemistry, 249, 7130-7139. 2. Kaymak, Ç., 2007. PhD Thesis, Gazi 9. Bradford, M.M., 1976. Anal. Biochem., University, Institute of Health Sciences, 72, 248. Ankara, Turkey. 10. Beutler, E., 1984. Inc. Orlando, FL 3. Parl, F., 2005. Cancer Letters, 221, 123- 32887, London. 129. 11. Huang, F., Li, S., Lu, X., Liu, A., Du, G., 4. Hayeshi, R., Mutingwende, I., Shi, G., 2011. Phytother. Res., 25, 284- Mavengere, W., Masiyanise, V., 89. Mukanganyama, S., 2007. Food Chem. 12. Aksoy, M., Küfrevioglu, O. I., 2017. Toxicol., 45, 286-95. Toxin Rev. 36, 1–7. 5. Kolobe, D. and Sayed, Y., 2004. 13. Özaslan, M. S., Demir, Y., Aslan, H. E., Biochemical Journal, 382(2), 703-709. Beydemir, Ş., Küfrevioğlu, Ö.İ., 2018. J. 6. Altınpınar, Ş, E., 2005. Master Thesis, Biochem. Mol. Toxic. 32, 22047. Cumhuriyet University, Institute of Health Sciences, Sivas, Turkey. 7. Tuna, G., 2008. Master Thesis, Hacettepe University Institute of Health Sciences, Ankara, Turkey.
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S.19. CHROMOSOMAL ABNORMALITIES IDENTIFIED AT PRENATAL DIAGNOSIS
Ayfer Pazarbasi1, Osman Demirhan1, Umit Luleyap1, Lutfiye Ozpak1, Seda N. Ilgaz1, I. Nur Uslu1, Gamze Comertpay1, Gulsevinc Ay1, Sabriye Kocaturk-Sel1, Nilgun Tanriverdi1, Selim Buyukkurt2
1 Department of Medical Biology, Faculty of Medicine, Cukurova University, Balcali, Adana, Turkey. 2 Department of Obstetrics and Gynecology, Faculty of Medicine, Cukurova University, Balcali, Adana, Turkey.
INTRODUCTION account for 6-11% of all stillbirths and neonatal
deaths. Chromosomal abnormalities that are Prenatal diagnosis of fetal chromosomal compatible with life but cause considerable abnormalities is the most common indication for morbidity occur in 0.65% of newborns (3). There invasive prenatal testing. Amniocentesis is a very are many strategies available to screen for crucial diagnostic procedure for preventing the chromosomal abnormalities, including combined birth of genetically defective fetuses in order to test, triple test, quad test, integrated screen, decrease the prevalence of genetic diseases in stepwise sequential screen and contingent populations. This technique was first performed sequential screen (4,5,6,7). All of these in the 1880's for decompression of approaches provide an adjusted risk for Down polyhydroamnios. In the 1950's, amniocentesis syndrome and trisomy 18, but they do not for fetal chromosome analysis was initiated as exclude the possibility of an affected fetus, laboratory techniques for cell culture and because the test sensitivity is less then 100%. karyotype were developed (1,2). The first Therefore, amniocentesis is still the only reported applications were limited to fetal sex diagnostic test in current use that is valid for determination. Prenatal diagnosis of fetal diagnosis (4,5,6,7). The purpose of this study was chromosomal abnormalities is the most common to document the results of the second trimester indication for invasive prenatal testing. The genetic amniocentesis we performed and provide prevalence of chromosomal abnormalities in a further comparison. clinically recognised early pregnancy loss is greater than 50%. Fetuses with aneuploidy 60 The Congress Is Supported By Turkish Airlines
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MATERIAL AND METHODS fetuses that were karyotyped; 3804 fetuses were males and 3763 fetuses were females and 569 Cytogenetic analysis and findings fetuses (6.99%) had various chromosomal A retrospective review of our amniocentesis abnormalities. So the rate of male/female was database for the period from January 2000 to 1.01. The group of advanced maternal age had September 2018 was carried out. The the highest rate of chromosomal abnormalities karyotyping of 8129 fetuses was carried out in and followed by positive triplescreening. Department of Medical Biology from the Chromosomal abnormalities associated with samples of amniotic fluids which were sent from maternal age. 55.53% of abnormal karyotypes Department of Gynecology and Obstetrics of (316 cases) were numerical and 42.17% (240 Balcali Hospital. A standart nomenclature has cases) were structural. Both numerical and been developed to describe each of types of structural chromosomal aberrations were abnormality found in human chromosomes. The observed in 13 cases (2.28%) (Figure 1). The current version was developed by the numerical abnormalities were as: trisomy 21 International Standing Committee on Human (47.78%), trisomy 18 (18.03%), monosomy X Cytogenetic Nomenclature and adopted in 1995 (10.12%), trisomy 13 (6.96%), triploidy (5.06%), (ISCN, 1995). Karyotypes of fetuses were Klinefelter Syndrome (3.48%), trisomy X performed on fetal cells that were obtained from (1.26%), XYY Syndrome (0.94%), mosaics and the amniotic fluid sample by using a long term the others (Figure 2). The frequent structural cell culture. After adequate growth, cultures were abnormalities were as: 46,XX/XY, inv(9) harvested in situ after an average 8 to 10 day (p11;q12) / (p11;q13) (31.25%), 46,XX / period. Karyotyping was routinely performed by XY,1qh(+) (10.41%), 46,XX / XY,16qh(+) G-banding by using the trypsin-giemsa staining (6.25%), 46,XY, Yqh(-) (6.25%), 46,XX / technique. At least 20 metaphases were analyzed. XY,9qh(+) (4.58%) and 46,XY, Yqh(+) (4.58%) A total of 8129 amniocentesis specimens were (Figure 3). Balanced and unbalanced processed during the study period. The mean translocations, deletions and duplications were maternal and gestational age of pregnant women also contributed to structural chromosomal evaluated for prenatal diagnosis were 31 years of abnormalities in lesser extent. age and 18.5 months respectively. Among 8129
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DISCUSSION AND CONCLUSIONS diagnosis is essential if there are recurrent Evaluation of the severity and etiology of the miscarriages for couples with a known anomaly is an important prognostic for prenatal chromosome chromosome rearrangement and if diagnosis (7). Abnormal karyotype composition there is advanced maternal age. Maternal age is is different according to different maternal the only well-established risk factor for Down amniocentesis indications. There is a variety of Syndrome, and the associated risk increases abnormal karyotypes in the second trimester exponentially at age 35 years and over (8,9,10). pregnancy, and the risk of fetal malformation is As a result, amniocentesis is regularly related with the kind of abnormal karyotype. Our recommended for women at age 35 and over. It data indicate that maternal serum triple marker is also recommended that case reports of screening is an effective method for detecting clinically normal subjects with balanced fetal chromosomal abnormalities in Turkish karyotypes should be published so that an women. It is also an effective screening tool for informed decision can be made prenatally when non-Down chromosomal abnormalities. a similar rearrangement is identified. In the rare Therefore, this screening method gives the group of parents carrying a balanced advantage of early counseling and of diagnosis as chromosome rearrangement affecting whole or early as in the second trimester of pregnancy. All partial 21q, the risk of having a child with a women in whom serum screening indicates an complete or partial trisomy 21 is relatively high, increased risk for fetal chromosomal with about 20.0% in females and 10.0% in males. abnormalities should be offered genetic If the translocation chromosomes and their counseling and chorionic villus sampling or normal homologous show pairing difficulties in amniocentesis. meiosis I, the additional risk of a 3:1 segregation It is generally accepted that balanced has to be taken into account, leading to a rearrangements lead to increased non disjunction recurrence risk of up to 30.0% (11). Carriers of of other chromosomes during meiosis. Carriers reciprocal translocations including chromosome of balanced translocations may be apparent 21 are at increased risk of having offspring with because of recurrent miscarriages with or without trisomy 21. Thus, the translocation could healthy and/or affected children (7). Prenatal promote formation of trisomy 21 in the offspring.
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Recent studies quite clearly indicate that environmental factors can increase the risk of interchromosomal effects (ICEs) do exist (12, fetal chromosomal abnormalities. Fetal 13-15). MD de Groot- van der Mooren et al. chromosomal abnormalities representing 6.99% wanted to estimate the livebirth prevalence of in our study group is crucial and underlines the T21 for the period of 2000–2013 in the importance of prenatal diagnosis for healthier Netherlands (16). A national screening pragnancies. programme was introduced for all pregnant REFERENCES women in 2007, which may have changed the livebirth prevalence of T21. There is no registry 1. Pazarbasi A, Demirhan O, Tastemir D, Tunc system regarding the livebirth prevalence of E, Ozgunen FT, Alptekin D, Evruke C, Demir C, trisomy 21 (T21) in the Netherlands. According Kasap M, Karakan Karakas Z, Inandiklioglu, to the study of MD de Groot- van der Mooren et Ozpak L. Evaluation of the cytogenetical results al. (2018) the proportion of mothers aged ≥ 36 of 4707 cases diagnosed with amniocentesis. years has increased from 12.2% in 2000 to 16.6% Cukurova Medical Journal, 2011; 36(1-4): 8-14. in 2009, to gradually decrease afterwards to (Article in Turkish) 15.2% in 2013 in Netherlands. The number of 2. Arikan I, Harma M, Barut A, Harma MI, Bayar tests performed adjusted for total livebirths U. Fetal loss rates after mid-trimester decreased (5.9% in 2000 vs. 3.2% in 2013) with amniocentesis. Health, Vol.2, No.4, 315-317 0.18% a year. Following invasive testing, a (2010): 315-317. higher proportion of foetuses was diagnosed with 3. American College of Obstetricians and T21 (1.6% in 2000 vs. 4.8% in 2013) with a Gynecologists. Invasive prenatal testing for significant increase of 0.22% a year. The aneuploidy. ACOG Practice Bulletin.No: 88, proportion of ToP (termination of pregnancy) Obstetrics and Gynecology, 2007, 110: 1459- subsequent to T21 diagnosis was on average 1467. 85.7%. This resulted in a stable T21 livebirth prevalence of 13.6 per 10,000 livebirths (16). 4. Karaoguz MY, Bal F, Yakut T, Ercelen NO, Ergun MA et al. Cytogenetic results of Corollary to literature and our findings revealed amniocentesis materials: incidence of abnormal that the advanced maternal age and certain
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karyotypes in the Turkish collaborative study. factors for Down Syndrome. Am J Hum Genet. Genetic Counseling, 2006; 17(2): 219-30. 2000; 67(3): 623-630.
5. Zhang YP, Wu JP, Li XT, Lei CX, Xu JZ, Yin 10. Hultén MA, Patel S, Jonasson J, Iwarssom E. M. Karyotype analysis of amniotic fluid cells and On the origin of the maternal age effect in comparison of chromosomal abnormality rate trisomy 21 DS: the Oocyte Mosaicism Selection during second trimester. Zhonghua Fu Chan Ke model. Reproduction. 2010; 139(1): 1-9. Za Zhi, 2011, 46(9): 644-8. (Article in Chinese). 11. Eggermann T, Schwanitz G. Genetics of 6. Ozalp Yuregir O, Buyukkurt S, Koc F, Down syndrome. In: Dey S, Ed. Genetics and Pazarbasi A. Prenatal Diagnosis. Archives Etiology of Down Syndrome, Part 1. Rijeka, Medical Review Journal, 2012, 21(1): 80-94. Croatia: InTech. 2011: 5-22. (Article in Turkish) 12. Kovaleva NV. Trisomy 21 in the offspring of 7. Demirhan O, Pazarbasi A, Guzel AI, Tastemir carriers of balanced non-contributing autosomal D, Yilmaz B, Kasap M, Ozgunen FT, Evruke C, rearrangements: examination of Demir C, Tunc E, Kocaturk-Sel S, Onatoglu- interchromosomal effect and non-homologous Arıkan D, Koc S, Ozer O, Inandiklioglu N. The meiotic co-orientation. In: van den Bosch A, reliability of maternal serum triple screening for Dubois E, Eds. New Developments in Down the prenatal diagnosis of fetal chromosomal Syndrome Research. New York, USA: Nova abnormalities in Turkish women. Genetic Science Publishers Inc. 2012: 149-176. (ISBN: Testing and Molecular Biomarkers; October 978-1-62081-893-0. Available from: 2011; 15(10): 701-707. https://www.novapublishers.com/ catalog / product_info.ph;? products_id=376). 8. Hassold TJ, Jacobs PA. Trisomy in man. Ann Rev Genet. 1984; 18: 69-97. 13. Anton E, Vidal F, Blanco J. Role of sperm FISH in the genetic reproductive advice of 9. Hobbs CA, Sherman SL, Yi P, Hopkins SE, structural reorganization carriers. Hum Reprod. Torfs CP, Hine RJ, et al. Polymorphisms in genes 2007; 22(8): 2088-2092. involved in folate metabolism as maternal risk
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14. Gianaroli L, Magli MC, Ferraretti AP, Dunne 16. Maurike D. de Groot- van der Mooren, S, Balicchia B, Escudero T, et al. Possible Saskia Tamminga, Dick Oepkes, Michel E. interchromosomal effect in embryos generated Weijerman, Martina C. Cornel. Older mothers by gametes from translocation carriers. Hum and increased impact of prenatal screening: Reprod. 2002; 17(12): 3201-3207. stable livebirth prevalence of trisomy 21 in the Netherlands for the period 2000–2013. European 15. Pujol A, Benet J, Staessen C, van Assche E, Journal of Human Genetics (2018) 26:157–165. Campillo M, Egozcue J, et al. The importance of aneuploidy screening in reciprocal translocation carrier. Reproduction. 2006; 131(6): 1025-1035.
Figure 1. Both numerical and structural anomalies in same karyotypes and their frequencies in 13 case.
Both numerical and structural anomalies in same karyotype Number (%) 47,XY +21, t(12;16)(q24;q24) 1 (7.69) 46,XX, t(14;21) +21 1 (7.69) 47,XY +18 t(1;18)(q31;q23) 1 (7.69) 69,XXY, 11p(+) 1 (7.69) 47,XY+21 inv(9) (p11;q13) 1 (7.69) 69,XXX, inv(9) 1 (7.69) 45,X0, inv(9) (p12;q13) 1 (7.69) 45,XY, t(14;15)(q10;q10) 1 (7.69) 45,XX, t(13;14) 1 (7.69) 45,XX, t(9;21) (p24;p11) 1 (7.69) 45,X0, del(10) 1 (7.69) 46,XY, inv (9)/45,X %30 1 (7.69) 46,XX, %10 different abnormalities 1 (7.69) Total frequency in all anomalies for both numerical and 13 /569 structural anomalies (2.28)
Figure 2. The frequencies of each numerical abnormalities all numerical abnormalities.
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Type of Numerical abnormalities Number (%)
47,XX/XY+21 151 (47.78)
47,XX/XY+18 57 (18.03) 45,X0 32 (10.12) 47,XX/XY+13 22 (6.96) 69,XXX/XXY 16 (5.06)
47,XXY 11 (3.48) 47,XXX 4 (1.26)
47,XYY 3 (0.94)
46,XX/47,XX+21(%40) 3 (0.94)
92,XXXX/ XXXY 2 (0.63) 46,XX/(Tetraploidy) %40 2 (0.63) 46,XY/47,XY+M(%50) 2 (0.63) 46,XY/(Aneuploidy) %11.5 1 (0.31)
53,XXXXX 1 (0.31) 47,XY+22 1 (0.31)
47,XY+M 1 (0.31)
46,XX/47,XXX(%8) 1 (0.31) 46,XY/47,XXY(%20) 1 (0.31) 46,XY/47,XY+2(3/25) 1 (0.31) 46,XX(%65)/46,X,-X+M(%40) 1 (0.31)
46,XY/47,XY+17(4/87) 1 (0.31)
46,XY/47,XX+13(%50) 1 (0.31) 46,XY/47,XXYY(%50) 1 (0.31)
46,XY(%70)/48,XXXY(%30) 1 (0.31) Total 316 (%55.53)
Figure 3. The frequencies of each structural abnormalities in whole (240) structural abnormalities.
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Type of structural abnormalities Number (%)
46,XX/XY, inv(9)(p11;q12), (p11;q13) 75 (31.25)
46,XX/XY 1qh+ 25 (10.41)
46,XY, Yqh(-) 15 (6.25)
46,XX/XY 16qh+ 15 (6.25)
46,XX/XY 9qh+ 11 (4.58)
46,XY, Yqh(+) 11 (4.58)
46,XX/XY 15p+ 5 (2.08)
46,XY 21p(+) 3 (1.25)
46,XY, 22s(+) 3 (1.25)
46,XY/XX 13p(+) 2 (0.83)
46,XY/XX, del 4p(15 →ter) 2 (0.83)
45,XX,rob t(13;14) (q11;q11) 2 (0.83)
46,XY rob t(13;13) (q11;q11) 2 (0.83)
46,XY, 21s(+) 2 (0.83)
46,XX, t(4;5)(p16;q31) 1 (0.41)
46,XX, t(7;15)(p11.2;p11.2) 1 (0.41)
46,XX(%90)/46,XX,t(3;9)(q25;qter)(%10) 1 (0.41)
46,XX t (4;9) (p16;q33) 1 (0.41)
46,XX t (6;9) 1 (0.41)
46,XYrob t (13q;21q)+21 1 (0.41)
46,XX t (12;20)(q24;q11) 1 (0.41)
46,XX t (5;7) 1 (0.41)
46,XY t (3;12) (q29;q23.1) 1 (0.41)
46,XY t(1;9) (p34;q34.3) 1 (0.41)
46,XY, der(5) t(5;7) (p15;p11) 1 (0.41)
46,XX der (7) t(3;7)(p11;q22) 1 (0.41)
46,XX, rob t(14;21) 1 (0.41)
46,XY der(22)rob t(21;22) (q10) 1 (0.41)
46,XY t (3;6) (q21;q15) 1 (0.41)
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46,XX t(3;9)(q21.1;p24.3) 1 (0.41)
46,XX t(5;10) (q35;q22.3) 1 (0.41)
46,XY t(9;13)(q11→ter;p11→9 tet) %50 1 (0.41)
46,XY, der(19), t(19;Y)(q11;Ypter) 1 (0.41)
46,XX (%85)/ 46,XX, t(5;20)(p13;p11)(%15) 1 (0.41)
46,XX(%90)/46,XX,t(3;9)(q25;qter)(%10) 1 (0.41)
46,XX inv (X)(p11;q12) 1 (0.41)
46,XX %5 chrom. abnormalities 1 (0.41)
46,XX del (13) p(11.2)ter 1 (0.41)
46,XY dup(Y)(p11.22→ter) 1 (0.41)
46,XY, 1qh(-) 1 (0.41)
46,XY 14s(+) 1 (0.41)
46,XY 15s(+) 1 (0.41)
46,XY del (12)(q13;q15) 1 (0.41)
46,XX 4p(+) 1 (0.41)
46,XY del(6)(q24.2-ter) 1 (0.41)
46,XY del(6)(q24-ter)(%12) 1 (0.41)
46,XX, fra22q 1 (0.41)
46,XX, fra17q 1 (0.41)
46,XY, fra14(q24) 1 (0.41)
46,XY, 9qh(+), t(1;14) 1 (0.41)
46,XY 22p(+) 1 (0.41)
46,XY,del(12)(q13;q15) 1 (0.41)
46,XY, 1qh(+), inv(9)(p13;q13) 1 (0.41)
46,XX, inv(1)(p13;q23) 1 (0.41)
46,XX, (%65)/46,X, fraXp?(%35) 1 (0.41)
46,XX 1qh+, 19q(+) 1 (0.41)
46,XX 2q(+) 1 (0.41)
46,XY 1qh+, Yqh(-) 1 (0.41)
46,XX, del 5p(14.3 →ter) 1 (0.41)
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46,XX/46,XY 1 (0.41)
46,XX 5p(+) 1 (0.41)
46,XY, fra5q(31) 1 (0.41)
46,XY, fra5q(31) (X2)(%20) 1 (0.41)
46,XX, dup(13)(q32 →ter) 1 (0.41)
46,XX, dup(5)(p13 →15) 1 (0.41)
46,XX, del 5p(13 →ter) (%10) 1 (0.41)
46,XX, t(8;9)(q22;q24) 1 (0.41)
46,XY, fraX(q27) 1 (0.41)
46,XX, (+)ace fra(3/25) 1 (0.41)
46,XY del(11)(q23) 1 (0.41)
46,XY, 22s(-) 1 (0.41)
Multiple abnormalities 1 (0.41)
Total 240 (42,17)
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S.20. HEMATOPOIETIC STEM CELL TRANSPLANTATION RESULTS IN PRIMARY IMMUNE DEFICIENCY DISEASES: THE EXPERIENCE OF ACIBADEM UNIVERSITY ADANA HOSPITAL
Barbaros Sahin Karagun
Division of Pediatric Hematology and Oncology, Faculty of Medicine, Acibadem University, Adana, Turkey.
Objectives: Primary immunodeficiencies are a Results: The median age of the patients was 9 rare group of diseases that have clinical and months. Patients were diagnosed with severe epidemiological importance due to leading high combined immunodeficiency (n= 6), Kostmann's morbidity and mortality. Hematopoietic stem cell syndrome (n = 1), MHC class-2 deficiency (n = transplantation is a curative treatment for many 4), Griscelli syndrome (n = 1), hemophagocytic combined primary immunodeficiency and lymphohistiocytosis syndrome (n = 1) and phagocytic disorders. Wiskott-Aldrich syndrome (n = 1). The results Methods: In this study, 18 children with primary didn't show graft failure. 17 patients survived, immune deficiency who underwent their follow -up and treatments are ongoing. One hematopoietic stem cell transplantation were patient died due to sepsis after transplantation. analyzed retrospectively. SPSS 17.0 package Conclusions: Early diagnosis by screening of program was used for statistical analysis. newborns for primary immunodeficiency Categorical measurements were expressed as diseases may lead to administration of number and percentage, and continuous hematopoietic stem cell transplantation on time measurements as mean and standard deviation. and can be survival of patients.
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S.21. EFFECTS OF BONE GRAFT MATERIAL AND OXIDATIVE STRESS ON FIBROBLASTS AND PROTECTIVE ROLE OF RESVERATROL
Kaleci B1,2, Koyutürk M1
1 Istanbul University-Cerrahpasa, Cerrahpasa Medical Faculty, Department of Histology and Embryology, 2 Institute of Health Science. Istanbul, Turkey.
Objectives: Bone grafts are most favorite were observed in RES and G combinations materials to support new bone formation. Ti- groups. Furthermore decreased cell proliferation oss®-graft-(G) as xenograft has been used index by H2O2 was stabilized with RES recently. We aimed to investigate the effects of application. There were no increases in OS levels G and oxidative stress-(OS) on 3T3-fibroblasts in G, RES and G+RES groups. RES application and potential protective role of resveratrol-(RES) decrease OS levels. H2O2+G+RES group in in-vitro wound healing model. Methods: In- indicated the highest COL1A2 expression. All vitro experiments were determined after 48- groups showed good migration quality except hours incubation, Cell proliferation was assessed H2O2 and H2O2+G groups. by population doubling time-(PDT), Conclusions: We concluded that Ti-oss® graft is Bromodeoxyuridine-(BrdU) and OS levels was biocompatible with fibroblasts; RES evaluated by immunofluorescence staining and administration decreases OS levels and improves all values were calculated by Image-J program. the healing process by increasing proliferation In addition, the expression of COL1A2-collagen index, migration and collagen synthesis. was evaluated by RT-PCR and 24 hour-cell This study was supported by the Research Fund migration was calculated in all experimental of Istanbul University. Project No. 21032 groups. Keywords: Bone graft, Oxidative stress, Results: Cell PDT index increase in H2O2 Resveratrol, Fibroblast, In-vitro incubation and decrease with RES combination. The highest levels of BrdU-immunoreactivity
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S.22. EFFECT AND RELIABILITY OF TRANSCRANIAL MAGNETIC STIMULATION ON PAIN IN STROKE PATIENTS
Türkan Turgay1, Serdar Akaltun2, Neytullah Turan2, Ozlem Altindag2
1Sanko University Research Hospital, Department of Physical Medicine and Rehabilitation, Gaziantep- Turkey. 2Gaziantep University Research Hospital, Department of Physical Medicine and Rehabilitation, Gaziantep-Turkey. Objectives: In this paper, we aimed to share the time stroke, subacute or chronic lesion and no effect of Transcranial Magnetic Stimulation other neurological involvement were included. (TMS) on central pain in patients with stroke. The exclusion criteria were as follows; multiple Materials and methods: 10 patients who had a lesions on cranial imaging, head trauma, acute stroke history were included in this study. stroke, epilepsy in their medical history. Patients with middle cerebral artery lesion, first- Pregnant women were also excluded.
Table 1. Demographic features in stroke patients
Duration of disease(year) 1 (min) 7 (max) 2.5 ± 1.7 (mean ± SS) Affected side 6 (right) 4 (left) 60/40 (ratio) Age 29 (min) 75 (max) 58.2 ± 16.1 Sex 8 (male) 2 (female) 80/20 (ratio)
Patients were examined before and after the first depressive mood was assessed by the Beck month of treatment with pain scale; Visual Depression Inventory (BDI). All patients Analog Scale (VAS), neuropathic pain level; received the current treatments and appropriate Leeds Assessment of Neuropathic Symptoms neurorehabilitation recommended for the and Signs Scale (LANSS) and Douleur treatment of ischemic stroke. Neuropathique en 4 Questions (DN4) and
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Results: Interestingly, our cases improved in pain.
Table 2. Comparison of clinical parameters in stroke patients.
Before Treatment After Treatment p LANSS 9.4 6.0 < 0.001 VAS (pain) 4.8 3.7 < 0.001 DN4 4.4 2.9 0.001 BDI 17.2 13.6 < 0.001
Conclusion: The neurophysiological mechanism hemispheres and to support functional healing. of functional recovery or disability is still Applying the TMS to the motor cortex may cause unclear. TMS applied over the primary motor small, short-term reductions in pain; but the cortex, stimulation of corticospinal tracts can results in the literature are limited. lead to presenting a motor evoked potential in the Key words: Transcranial magnetic stimulation, contralateral extremity with painless technique. stroke, pain. TMS aims to rebalance communication between
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S.23. THE EFFECT OF BLOOD PRESSURE CUFF PROTECTIVE SHIELD TO PREVENT POSITION-RELATED COMPLICATIONS DURING THYROID AND PARATHYROID SURGERY
Betül Kocamer Şimşek1, Göktürk Maralcan2
1Sanko University Research Hospital, Department of Anesthesia, Gaziantep-Turkey. 2Sanko University Research Hospital, Department of General Surgery, Gaziantep-Turkey.
Thyroid surgery is the most commonly it difficult for the anesthesia team to manage the performed endocrine surgery and the special patient. Efforts to rearrange the cuff disrupt the thyroid position is given in order to perform the operation and affect concentration. surgery in the best conditions. The surgical We have been using blood pressure cuff access warrants maximum exposure of thyroid protective shield produced with hard PVC which gland which can be achieved by placing a padded is seen in the picture 1, in Gaziantep University ring under the head of the patient and a rolled and with the same team in Sanko University, Sani sheet under the shoulders. During surgery, one or Konukoğlu Training Hospital for nine years in two of the patient's arms are taken to juxtaposed over 400 patients to prevent the problems to patient in order to the surgical team to work mentioned before with taking the both arms are comfortably. When one arm is juxtaposed to juxtaposed to patient. Owing to this blood patient the other arm positioned in 900 abduction pressure cuff protective shield we did not observe but during the surgery both surgeon or assistance brachial plexus injuries or any incorrect team’s pushing the arm makes it over 900 interventions due to incorrect blood pressure abduction and consequently this can cause nerve measurements. These data are preliminary results injuries especially brachial plexus injuries due to of our study. nerve stretching. In order to prevent this, when We consider that due to this low cost and simple, both arms are juxtaposed to patient, incorrrect at the same time functional protective shield blood pressure measurement can be seen due to complications that may occur due to the patient's pressure on the blood pressure cuff. This makes position will decrease and this shield will prevent
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incorrect blood pressure measurements and the operation will not deteriorate.
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S.24. EXPERIENCE OF AN INNOVATIVE CYTOKINE APHERESIS METHOD IN SEPTIC SHOCK: A RETROSPECTIVE ANALYSIS OF 26 PATIENTS AT A REANIMATION ICU
Berna KAYA UGUR1, Mehmet YILMAZ2, Suleyman GANIDAGLI1, Pinar TUMTURK1 Ibrahim OZASLAN1, Seval KUL3, Gokhan GOKASLAN4
1 Gaziantep University Department of Anesthesiology and Reanimation, Gaziantep, Turkey. 2 SANKO University Department of Hematology, Gaziantep, Turkey. 3 Gaziantep University Department of Biostatistics, Gaziantep, Turkey. 4 SANKO University Department of Cardiovascular Surgery, Gaziantep, Turkey.
Objectives: Sepsis is the primary cause of death Methods: Patients with septic shock that for patients in the intensive care unit (ICU) (1, 2). received Cytosorb therapy at the ICU of Despite meticulous efforts for treatment and Department of Anesthesiology and Reanimation follow up; mortality rates of patients with sepsis and Cardiovascular Surgery Gaziantep still remains high. There is limited published data University School of Medicine, from January 1st in the literature up to date concerning clinical 2015 to August 1st 2015 were retrospectively experience of the procedure in patients with reviewed from hospital records after ethical sepsis. As far as we know this is the first study approval was obtained from Gaziantep reporting efficacy of Cytosorb therapy on a series University, Ethical Committee. Diagnosis of of patients with septic shock. We aimed to assess septic shock was made according to the classic the effect of Cytosorb on inflammatory acute criteria (3). Ages, gender, body mass index phase reactants (C-reactive protein, (BMI) of the patients were obtained. Laboratory procalcitonin), alanine transaminase (ALT) and tests including CRP, procalcitonin, ALT, creatinine levels and mortality rates in this creatinine, bilirubin, ESR, APACHE II scores retrospective study. and inotropic indicis, number of Cytosorb therapy courses and clinical outcomes were all
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recorded before and after each course of second and third courses, this difference was not Cytosorb treatment. Inotropic indicis of the statistically significant (p=0.465, 0,533; patients were calculated with the formula defined respectively). Similarly, procalcitonin levels by Wernovsky(4). Kolmogorov-Smirnov test were significantly lower after the first course was used to check the normality of distribution of (p=0.045) and insignificantly lower after the continuous variables. Paired samples T test was second and third courses (p values were 0.795 used for comparison of dependent variables with and 0.386; respectively). There was a slight and a normal distribution and Wilcoxon test was used insignificant decrease in ALT levels after the first for comparison of dependent variables with an course (p=0.186), but the decrease was abnormal distribution. SPSS for Windows ® significant after the second and third courses (p version 22.0 was utilized for statistical analysis. values were 0.016 and 0.043; respectively). A p value below 0.05 was considered as There was no significant difference in creatinine significant. levels after the first, second and third courses (p Results: There were 26 patients with septic values were 0.366, 0.305 and 0.564; shock at the ICU and received Cytosorb therapy respectively). There was a significant difference (19 male, 7 female) in the hospital records. The in sedimentation rates after the first course average age and BMI of the patients was (p=0.014). But there was no difference in 50.46±20.53 years (min. 17, max. 84) and sedimentation rates after the second and third 26.15±5.11 kg/m2 (19.7 min, 36.7 max); courses (p values were 0.180 and 0.769; respectively. Average number of performed respectively). Also there was no significant Cytosorb procedures was 4.12±2.97 (min. 1, difference in WBC levels after the first, second max. 10). The procedures were performed at least and third courses (p values were 0.190, 0.153 and twice in 21 patients and at least three times in 15 0.195; respectively). There was no significant patients. CRP, procalcitonin, ALT and creatinine difference in inotropic indicis of patients after the (CRE) levels of patients are presented in Table 1. first, second and third courses (p values were There was a significant decrease in CRP values 0.906, 0.786 and 0.931; respectively). There was after the first course of Cytosorb procedures no significant difference in bilirubin levels after (p=0.004). Although there was a decrease after the first and second course (p values were and
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third courses (p values were 0.089 and 0.454; respectively) but there was no difference after the respectively) but bilirubin levels were third course (p=0.834”). Two of the 26 patients significantly increased after the third course with septic shock that have received Cytosorb (p=0.048). There were significant differences in therapy eventually recovered and were Apache II scores after the first and second discharged from ICU and 24 have died. courses (p values were 0.002 and 0.006;
Table1. Laboratory test values of patients that received Cytosorb therapy.
Procedur CRP Procalcitonin ALT Creatinine e
Before After Before After Before After Befor After e First 169,0 22,54 154,9 154,5 2,35 2,23 193,5 29,00 ± ± 6±242 4±242 ±1,4 ±1,3 4±103, ± (n=26) 96,49 31,07† ,78 ,76 1 4 85* 38,46† * Second 158,10 156,24 16,76± 17,81± 149,95 129,86 1,90± 1,71± ± 93,32 ± 82,43 22,93 25,11 ± ± 1,34 1,01 (n=21) 263,13§ 227,82§ Third 149,46 146,62 23,36± 22,93± 99,13± 76,13± 1,64± 1,71± ± 79,87 ± 88,39 29,10 32,94 146,32∆ 99,91∆ 1,01 0,91 (n=15) *p=0.004, †p=0.045, §p=0.016, ∆p=0.043
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Procedur ESH WBC Inotropic Index e Before After Before After Before After First 43,42 36,46 11,52 10,49 86,52 83,50 ±21,0 ±17,8 ±9,28 ±8,17 ±85,8 ±80,6 (n=26) 7* 8* 0 0 Second 35,43± 39,67± 10,09± 8,70±6, 63,15± 65,92± 17,70 16,19 7,35 28 51,45 54,32 (n=21) Third 36,73± 37,73± 8,97±5, 10,48± 53,51± 54,76± 13,78 16,51 19 5,30 46,96 48,21 (n=15) *p=0.014
Procedure BILIRUBIN APACHE II SCORE Before After Before After First 2,00± 2,06 2,20± 2,11 20,81± 6,40† 23,12± 4,68† (n=26) Second 1,71± 1,06 1,86± 1,32 22,62± 4,84§ 25,29± 5,86§ (n=21) Third 1,91± 1,47* 2,33± 1,70* 23,80± 5,98 23,80± 6,76 (n=15) *p=0.048, †p=0.002, §p=0.006
Conclusions: The mortality rate of patients with eventually generates multi-organ injuries (5). septic shock is still high and one of the most Despite numerous efforts and challenges there is challenging issues at the ICUs (1, 2). Systemic still an urgent need for a therapeutic modality to inflammatory response to infection or injury is reverse, or at least alleviate the multi-organ believed to be the part responsible for damage. development of sepsis. Subsequent release of numerous cell-associated and soluble molecules 79 The Congress Is Supported By Turkish Airlines
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Extracorporeal blood purification therapies have (C-reactive protein, procalcitonin) after the first been used in sepsis as an adjuvant treatment to course of Cytosorb therapy, but this decrease was improve outcomes. CytoSorb ® (CytoSorbents not significant after further courses. This may be Corporation, USA) is a novel sorbent due to the effective removal of excess hemoadsorption device for cytokine removal. inflammatory response products after the first There are only a number of case reports and course and limited effect thereafter. Interestingly, experimental studies evaluating the effects of improvement of liver functions roughly assessed Cytosorb in cytokine levels and inflammatory by ALT was observed not after the first but the markers in the literature. However there is second and third courses. This may be probably scarcity of data concerning clinical outcome of linked to lowering detrimental cytokines and Cytosorb treatment in patients with septic shock. other end products of systemic inflammation that Wiegele reported improvement of renal functions results in liver injury. Although we could not and liver enzymes in a patient with Legionella reveal a significant improvement in creatinine pneumonia that received Cytosorb treatment (6). levels that gives clue about renal function, Also, Mitzner et al. reported decreased levels of decreasing levels may be promising. Three out IL-6, CRP, creatinine, procalcitonin, leukocyte of 26 patients recovered from septic shock. levels in an 80 year old chronic hemodialysis Unfortunately one of the patients that recovered patient with septic shock after Cytosorb added to died of complications due to coagulation hemodiafiltration (7). In animal models with abnormalities, other two were uneventfully septic shock, CytoSorb therapy was associated discharged from the ICU. with reduced inflammation and improved In the present study CRP, procalcitonin and ALT outcomes (8). levels were decreased after the Cytosorb courses, There is no sufficient data regarding Cytosorb but there was no significant difference in therapy in patients with septic shock and to our creatinine levels. However these decreased knowledge this is the first report regarding results inflammatory parameters and enzyme levels did of Cytosorb therapy in a patient series in the not improve the overall survival. There is evident English literature. We observed a similar need for further large scaled studies to decide the decrease of inflammatory acute phase reactants timing, number of procedure, efficacy and
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feasibility of this novel approach in septic shock severe sepsis and septic shock, 2012. Intensive management that still has unfavorable outcomes. Care Med. 2013 ;39:165-228. 4. Gaies MG, Gurney JG, Yen AH, Napoli ML, References: Gajarski RJ, Ohye RG, Charpie JR,Hirsch JC. Vasoactive-inotropic score as a predictor of 1. Angus DC, Linde-Zwirble WT, Lidicker J, morbidity and mortality in infants after Clermont G, Carcillo J, Pinsky MR. cardiopulmonary bypass. Pediatr Crit Care Med. Epidemiology of severe sepsis in the United 2010. States: analysis of incidence, outcome, and 5. Mar;11(2):234-8.Bone RC. The pathogenesis associated costs of care. Crit Care Med. of sepsis. Ann Intern Med. 1991 Sep 15;115:457- 2001;29:1303-10. 69. 2. Vincent JL, Sakr Y, Sprung CL, Ranieri VM, 6. Wiegele M, Krenn CG. Cytosorb™ in a patient Reinhart K, Gerlach H, Moreno R, Carlet J, Le with Legionella pneumonia-associated Gall JR, Payen D; Sepsis Occurrence in Acutely rhabdomyolysis: a case report. ASAIO J. Ill Patients Investigators. Sepsis in European 2015;61:e14-6. intensive care units: results of the SOAP study. 7. Mitzner SR, Gloger M, Henschel J, Koball S. Crit Care Med. 2006;34:344-53. Improvement of hemodynamic and 3. Dellinger RP, Levy MM, Rhodes A, Annane inflammatory parameters by combined D, Gerlach H, Opal SM, Sevransky JE, Sprung hemoadsorption and hemodiafiltration in septic CL, Douglas IS, Jaeschke R, Osborn TM, shock: a case report. Blood Purif. 201;35:314-5. Nunnally ME, Townsend SR, Reinhart K, 8. Kellum JA, Song M, Venkataraman R. Kleinpell RM, Angus DC, Deutschman CS, Hemoadsorption removes tumor necrosis factor, Machado FR, Rubenfeld GD, Webb S, Beale RJ, interleukin-6, and interleukin-10, reduces Vincent JL, Moreno R; Surviving Sepsis nuclear factor-kappaB DNA binding, and Campaign Guidelines Committee including The improves short-term survival in lethal Pediatric Subgroup. Surviving Sepsis Campaign: endotoxemia. Crit Care Med. 2004;32:801-5. international guidelines for management of
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S.25. SINGLE NUCLEOTIDE POLYMORPHISM DETECTION OF IL7R (RS6897932) GENE IN MULTIPLE SCLEROSIS OF NORTHERN IRAQI POPULATION
Rozhgar A. KHAILANY1,2, Naser GILANI2, Khandakar A S M SAADAT2, Belan O. KANABE3, Mehmet OZASLAN3
1 Department of Biology, College of Science, University of Salahaddin, Erbil, Iraq. 2 Department of Medical Biology and Genetics, University of Gaziantep, Gaziantep, Turkey. 3 Department of Biology, Gaziantep University, Gaziantep, Turkey.
Multiple sclerosis (MS) is a chronic amplification refractory mutation system-PCR inflammatory demyelinating and (ARMS-PCR) technique. As a result of gene neurodegenerative disease of central nervous polymorphism screening, comparison between system with unknown causes. The risk of MS alleles and genotypes in the MS patients and involves both genetic and environment factors. healthy controls show significant differences (P The IL-7R is a promising candidate for MS, = 0.021). In conclusion, the key point of this because its involvement in the autoimmunity, study is that the rs6897932 SNP was significantly and regulating of the T-cell homeostasis, associated with an increased risk of MS in proliferation, and anti-apoptotic signaling. We patients living in the North of Iraq. However, aimed to investigate rs6897932 gene further studies on larger population from polymorphism in association with MS in different parts of the country are still required. northern Iraq population. The study included 180 Keywords: Multiple Sclerosis, IL7R Gene, samples; 89 control and 91 patient samples. Polymorphism, ARMS-PCR. Genomic DNA was isolated from blood specimens, the polymorphism analyzed by
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S.26. NEXT GENERATION SEQUENCING REVEALS ASSOCIATION OF c.T850C KCNQ2 GENE VARIANT WITH UNKNOWN SIGNIFICANCE CAUSED EARLY EPILEPTIC INFANTILE ENCEPHALOPATHY (EIEE): A CASE REPORT
Naser GILANI1, Rozhgar A. KHAILANY1,2, Khandakar A S M SAADAT2
1Department of Medical Biology and Genetics, University of Gaziantep, Gaziantep, Turkey. 2Department of Biology, College of Science, University of Salahaddin, Erbil, Iraq.
Early-onset epileptic encephalopathy, a severe completely moveless and bed rested, with condition than characterized by epilepsy and convulsion attacks controlled by anti-epileptic profound intellectual disability. The seizures medicines, hairy body skin, dental deformity, begin in the first weeks of life and typically show abnormal body posture like decelerate posturing, little response to treatment. Most affected and no defecation control. Vision and hearing individuals are unable to talk, and they have low loss, just has some response to sounds as jerking, muscle tone (hypotonia) or very stiff muscles, he is only child of family. On molecular causing difficulty with movement. Mutations in investigation, we did for proband whole exome many genes can lead to EIEE, one of them is sequencing in which all genes of human genome KCNQ2 (potassium voltage-gated channel are sequenced using next generation sequencing subfamily Q member 2). The case we are going (NGS), a technique revolutionized the way to present is a 6 year-old male child, product of a scientists look at medical genetics. Mutations in normal pregnancy with C/S at 8 months of 3 genes (KCNQ2, OTC, KIAA0586) were gestational age and normal prenatal screening reported. From these mutations, c.T850C on tests, from beginning generally he was hypotonic KCNQ2 gene was consistent with clinical and weak crying, impaired swallowing, two features of proband more than others. In second weeks after birth seizures attacks has begun, step we start carrier study through sanger currently he is mentally and physically retarded, sequencing for parents for the variant. The 83 The Congress Is Supported By Turkish Airlines
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parents were normal for the variant. It means the Keywords: Next Generation Sequencing, Early variant is most probably a de novo mutation, as Infantile Epileptic Encephalopathy, KCNQ2, de we expect for this Autosomal Dominant Disease. novo.
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S.27. INHIBITORY EFFECT OF SOME ANTIPSYCHOTIC DRUGS ON HUMAN GLUCOSE 6 PHOSPHATE DEHYDROGENASE: IN SILICO AND IN VITRO STUDY
Muhammet Karaman1, 2, Halide Sedef Karaman3, Mehmet Çiftci4
1Department of Molecular Biology and Genetics, Faculty of Arts and Science, Kilis 7 Aralik University, Kilis, Turkey. 2Advanced Technology Application and Research Center (ATACR), Kilis 7 Aralik University, Kilis, Turkey. 3Department of Chemistry, Faculty of Science, Atatürk University, Erzurum, Turkey. 4Department of Chemistry, Faculty of Arts and Science, Bingol University, Bingol, Turkey.
Objectives: Glucose 6 phosphate dehydrogenase from human red blood cells. Kinetic studies were (G6PD) enzyme catalysis the reaction, NADPH performed with antipsychotic drugs including is produced by oxidizing glucose 6 phosphate risperidone, haloperidol, aripiprazole, (G6P) into 6-phosphogluconolactone (6PG). The escitalopram, and piracetam. Following kinetic enzyme plays an important role in producing studies, molecular docking studies were reduced nicotinamide adenine dinucleotide performed to display inhibiting mechanism of phosphate (NADPH) and ribose 5 phosphate. drugs against hG6PD. However, gene of G6PD often mutates and Results: hG6PD enzyme was purified with 67% G6PD deficiency is a common genetic disorder yield from human red blood cells. In the kinetic throughout the world. People with G6PD studies, it was detected that escitalopram and deficiency be faced with intravascular, and piracetam has no effect on hG6PD activity, extravascular hemolysis in red blood cells due to however, aripiprazole activates the enzyme. On accumulating of reactive oxygen species. The the contrary, risperidone and haloperidol purpose of this project is to observe effect of inhibited the enzyme activity with 0.053 mM and some antipsychotic drugs on human G6PD. 0.153 mM of IC50 value, respectively. Moreover,
Methods: hG6PD enzyme was purified with 2’5’ it was detected that inhibition type of risperidone ADP Sepharose 4B affinity chromatography is noncompetitive inhibition on hG6PD.
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Conclusions: In this study, risperidone exhibited in vitro inhibitory effects on hG6PD activity. When risperidone use by a patient with G6PD deficiency, it can cause adverse side effects. Therefore, we suggested that risperidone should be carefully used.
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S.28. PREPARATION OF FE3O4- POLY(PHENYLBORONIC ACID) COMPOSITE FILM MODIFIED PLATINUM ELECTRODES FOR DETERMINATION OF GLUCOSE WITH DIFFERENCIAL PULS VOLTAMMETRY
Öznur Güngör
İnönü University, Faculty of Arts and Science, Chemistry Department, 44280, Malatya, Turkey.
Abstract Introduction
The present study reports the determination of Glucose is a six-carbon monosaccharide with a glucose in 0.10 M PBS (pH 6,00) using melting point of 146 0C. D-glucose is important electropolymerized ultrathin film of Fe3O4 dietary sources of energy from natural containing poly(phenylboronic acid) on platinum carbohydrate sources and is used as processed electrode. The electrochemical feature of the food additives [1]. Glucose can be found in the L modified electrode was characterized by cyclic and D conformation. But our body only voltammetry (CV). The electrochemical recognizes D-glucose which is the energy source behaviour of the biosensor was also investigated of choice for the brain[2]. High concentration of using differencial puls voltammetry. Segment glucose in the blood leads to diabetes, which is a cycle, scan rate and pH are optimized. The common disease today. Visual disturbances, different ratios of Fe3O4 are working. Thus, it has nerve damage, heart failure, late healing of been claimed that modified electrode can be used wounds may be observed in patients with successfully as a glucose selective membrane diabetes. Blood glucose should be continuously because of the easy preparation and very high R- monitored because of cause permanent damage value (0.9987). to organs or even death in the later stages. Blood glucose monitoring device is the detection Keywords: Fe3O4, phenylboronic acid, glucose, technology that measures the concentration of sensor, electropolymerization. glucose. Much effort has been made to improve the performance of this technology in the last 56
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years, since the invention of the first enzyme prepared electrodes were used glucose electrode in 1962 by Clark and Lyons [3]. Many determination. methods exist to measure the concentration of D- Experimental glucose including electrochemical and optical approaches. There are many reports on the Materials: Phenylboronic acid (PBA) was evolution of blood glucose sensors [4-6]. obtained from Chem- Impex International Inc. Phenylboronic acid (PBA) derivatives bearing 0.1 M phosphate buffer saline (PBS) (pH 6,00) redox-active moieties. PBA can be used for the was used as the supporting electrolyte solution. voltammetric determination of sugars, in which PBA solutions were prepared just before running the redox potential and/or current are dependent each experiment. Glucose and Fe3O4(50- 100 nm on the sugar concentration. Sugars and their particle size) were analytical grade and supplied derivatives can be detected in optical and either by Sigma-Aldrich Chemical Company. electrochemical by the help of boronic acid [7]. Glucose solution was prepared freshly for each The electrochemical method is a well-known measurement. Ultra pure water was optained method for the synthesis of conductive polymers from the Millipore brand Elix 20 model water with various techniques. In this method, the system. Pt electrode was mechanically cleaned monomer can obtain the polymer on the electrode using aqueous diamond pastes successively on surface or solution as a result of electrolysis by diamond polishing pads and alumina slurry on placing it in a polymerization cell together with alumina polishing pads. After each polishing step an appropriate solvent or support electrolyte. The modified electrodes were washed with distilled radical anion or radical cation chain formed by water and ultrasonicated about 2 minutes in an the reduction or oxidation of the monomer is ultrasonic bath (Branson model 3510). growing and these form the conductive polymer Instrumentation chains [8]. All voltammetric measurements were performed In this study, poly(phenylboronic acid)- Fe3O4 by using a conventional electrochemical cell with composite membranes on the Pt electrode a three-electrode system, comprising a platinum surfaces were prepared by CV technique. These disk electrode (CHI, 2 mm diameter) as the working electrode, a Ag/AgCl electrode 88 The Congress Is Supported By Turkish Airlines
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saturated with KCl as the reference electrode and electroanalytical technique with prepared a Pt wire coil as the auxiliary electrode. These modified electrode for the qualitative and electrodes were immersed into the 10 mL quantitative detection of glucose. electrochemical cell. CV and DPV Results and Discussion measurements were obtained from the BAS 100W (Bioanalytical Systems, Inc.) Voltammetric measurements with the electrochemical analyzer. The pH was measured modified electrodes with a Jenway 3010 pH meter. Measurements The polymeric films were grown on PtE by were carried out at room temperature. cyclic voltammetry. Many different segment cycles have been attempted. Films were grown Preparation of Fe3O4- poly (phenylboronic acid) electrodes for 5 segment cycles unless stated otherwise (Figure 1). Electrode was developed on poly(phenylboronic acid) (pPBA) modified platinum electrode (PtEs) for the measurement of glucose. Firstly, PtEs were modified with electropolymerization of the p- phenylboronic acid as a monomer by cyclic voltammetry (CV) in 0,1 M phosphate buffer saline (PBS) (pH 6,00) at -1000- 1000 mV . Then the electrochemical oxidation of PBA performed in 0.1 M PBS pH 6,00 on pPBA modified PtE. After all electrolyte solution which concentration ratio of 1:4 and 1:8 (phenylboronic acid:Fe O ), 3 4 Figure 1 The relationship between segment prepared in PBS. Then the electrochemical cycles and oxidation peak current for PBA on oxidation of Fe O - pPBA performed in 0.1 M 3 4 modified PtE (a=2 segment; b=3 and 8 segment; PBS pH 6,00 on modified PtE. The effects of c= 4 and 7 segment; d= 6 segment; e= 5 segment) various electrochemical parameters, such as pH and scan rate were examined. Differential pulse voltammetry (DPV) is used as an ideal 89 The Congress Is Supported By Turkish Airlines
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The pH influence of the 0.1 M PBS electrolyte on the voltammetric response of pPBA modifed electrode was investigated in the range pH 5–8. The maximum response was observed at pH 6,00 (Figure 2).
Figure 3 The relationship between scan rate and oxidation peak current for PBA on modified PtE in background solution.
A Pt electrode can be modified with pPBA by electrochemical reduction of the phenylboronic Figure 2 The relationship between electrolyte pH acid. A voltammetric scan of a Pt electrode in and peak current of PBA on modified PtE in 0.1 aqueous solution produced pPBA multilayers on M PBS. the surface of the electrode. Bare Pt electrode The influence of the scan rate on the DPV exhibited a linear response to glucose over the response for pPBA modifed electrode in the concentration range 1–10 nM/mL (Figure 4).The range of 50-120 mV/s was examinated. The PBA modified electrode exhibited a linear maximum response was found to be for 100 response to glucose over the concentration range mV/s scan rate in 0.1 M PBS at pH 6,00 (Figure 1–10 nM/mL (Figure 5) 3).
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B
Figure 5 (A) DPVs of the PBA modified Figure 4 DPVs of the Bare Pt electrode to electrode to increasing concentration of glucose, increasing concentration of glucose (B) Calibration curve of the PBA modified (concentration range 1–10 nM/mL) electrode for increase glucose concentrations. (concentration range 1–10 nM/mL)
A A Pt electrode can be modified with Fe3O4- poly(phenylboronic acid) by electrochemical reduction of the phenylboronic acid. A voltammetric scan of a Pt electrode in aqueous
solution produced Fe3O4-pPBA multilayers on the surface of the electrode. The pPBA modified electrode exhibited a linear response to glucose over the concentration range 1–10 nM/mL (Figure 6-7).
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A A
B B
Figure 7 (A) DPVs of the Fe3O4- Figure 6 (A) DPVs of the Fe O - 3 4 poly(phenylboronic acid) electrode to increasing poly(phenylboronic acid) electrode to increasing concentration of glucose, (B) Calibration curve concentration of glucose, (B) Calibration curve of Fe3O4-poly(phenylboronic acid) electrode for of the Fe3O4- poly(phenylboronic acid) increase glucose concentrations. (concentration electrode for increase glucose concentrations. ratio of 1:8 (phenylboronic acid:Fe3O4) (concentration ratio of 1:4 (phenylboronic acid:Fe3O4) 92 The Congress Is Supported By Turkish Airlines
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The magnitude and reproducibility of the DPV Boronic acid-modified electrodes have been responses obtained with eighteen measurements successfully used for the development of showed the stability of the modified electrode for electrochemical sensors for detection of 10 nM/mL glucose in background solution biomolecules becauseof the easy preparation and (Figure8). high R-value (0.9987).
References
[1] Heike S. Mader, Otto S. Wolfbeis. Microchim Acta (2008) 162: 1–34. [2] Olsan, A., Ullmann‟s Encyclopedia of Industrial Chemistry, VCH Publishers, New York, A. 12: 457-475 (1991). [3] Clark, L. C. & Lyons, C. 1962. Electrode systems for continuous monitoring in cardiovascular surgery. Ann Ny Acad Sci, 102, 29e45.
Figure 8 The reversibility of electrochemical [4] Park, S., Boo, H. & Chung, T. D. Anal. Chim. Acta, (2006) 556, 46-57. measurements for the glucose-based Fe3O4- [5] Yoo, E. H. & Lee, S. Y. Sensors, ( 2010) 10, poly(phenylboronic acid) modified electrode. 4558-4576.
Conclusions [6] Heller, A. & Feldman, B. Accounts Chem. Res., (2010) 43, 963-973. The optimum conditions for the analysis of PBA [7] Lin Liu, Ning Xia, Yun Xing, Dehua Deng. were found as 0.1 M PBS pH 6,00 and 100 mV/s Int. J. Electrochem. Sci., 8 (2013) 11161 – 11174. scan rate. Under the optimum conditions, the [8] Aydın, Y., “İletken Polimerlerin İçerisinde peak current was linearly related to the Enzim Tutuklamasıyla Yapılan Biyosensörler”, concentration of PBA in the range of 1–10 Yüksek Lisans Tezi, Karamanoğlu Mehmet Bey Üniversitesi Fen Bilimleri Enstitüsü, Karaman, nM/mL. Comparied with pPBA modified 2-14 (2012). electrode to Fe3O4 –pPBA modified electrode. Fe3O4 containing membrane electrodes were showed high linearity and good and R-value.
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S.29. EFFECT OF ENDOTRACHEAL TUBE CUFF PRESSURE ON DEVELOPMENT OF VENTILATOR ASSOCIATED PNEUMONIA
Nevhiz Gündoğdu1, Nevin uysal2, Öner Dikensoy3
1 Department of Pulmonary Medicine, SANKO University. 2 Assistant Professor, Division of Pulmonary and Critical Care Medicine, Medical College of Wisconsin. 3 Professor, Department of Pulmonary Medicine, Taksim Acıbadem Hospital.
Objectives: Ventilator associated pneumonia Results: 196 patients were intubated during the may cause prolonged hospital stay and mortality. study period. Of these patients twenty six The aim of this study was to detect the effect of reached inclusion criteria and randomised to endotracheal tube cuff pressure on ventilator groups of low-normal and high-normal associated pneumonia. endotracheal cuff pressures. Median cuff Methods: Patients who were intubated in ICU pressure at low-normal group was 18.2 (16.0- after admission in Gaziantep University Hospital 21.4) cmH2O and it was 22 (20.1-24.8) cmH2O from March 2008 to October 2008 were in high-normal group (p=0.02). There was evaluated. Immune suppression, malignancy, microbial growth in 75% of tracheal aspirate witnessed aspiration before intubation, cultures. Empiric antibiotic treatment was cerebrovascular accident and younger than 18 sufficient in only 12.5% of patients based on the years old patients were excluded from the study. results of antibiogram. The incidence of VAP Patients were randomised to low-normal and was 41.7% (64 in 1000 ventilator days) in the high-normal cuff pressures; prospectively. Cuff low-normal group and 21.4% (41 at 1000 pressures were set at 20 cm H2O or 30 cmH2O ventilatory day) in the high-normal group and measured once in every four hours and (p=0.27). All patients with VAP were died. adjusted if necessary. National Nosocomial Mortality rates between two groups were similar Infection Surveillance System criteria were used (p=0.56). for VAP diagnosis.
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Conclusion: There was no statistically significant difference between the two groups in the incidence of VAP or mortality.
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Poster Presentation Abstracts for the SANKO University Innovation in Medicine Summit-IV 11th-13th Oct 2018, Gaziantep, Turkey
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P.1. TUMOR-ENDOTHELIAL CROSS-TALK IN CO-CULTURE: CHANGES OF PROLIFERATION AND INVASION PROFILES
Zeynep Betül Sari1,2, Emine Yavuz2, Tülin Cora1
1Department of Medical Genetics, Selcuk University, Medical Faculty, Selcuklu, Konya, Turkey. 2Selcuk University Advanced Technology Research and Application Center, Selcuklu, Konya, Turkey.
Objectives: co-cultivation system to analyze the influence of The tumor microenvironment (TME) plays a key healthy cells on tumor cells, also the influence of role in proliferation, invasion, and metastasis of tumor cells on healthy cells. many solid cancers as in head and neck Methods: squamous cell carcinoma (HNSCC) which is the For indirect co-cultivation system, we prepared sixth most common cancer worldwide (1). TME conditioned medium (CM) from HUVEC cell is composed of cellular and acellular components cultures to mimic the environment of HUVEC including cancer-associated fibroblasts, immune cells around the tumor cells. HUVEC cells were cells, blood and lymphatic tumor vessels, the plated in DMEM containing 10% FBS and extracellular matrix, chemokines, cytokines, and allowed to attach overnight. The supernatants growth factors (2-5). Endothelial cells also were collected, centrifuged to remove cell debris, constitute an important part of HNSCC and called as HUVEC-CM to culture HEp-2 microenvironment (6). However, the underlying cells. We also prepared conditioned medium mechanisms of their functions within the (CM) from HEp-2 cell cultures to mimic the microenvironment remain poorly understood. environment of HEp-2 cells around the This study aims to investigate the impact of co- endothelial cells called as HEp2-CM to culture culture of laryngeal cancer cells (HEp-2) and HUVEC cells. Cells were cultured in the endothelial cells (HUVEC) on invasion and presence of increasing concentrations (%0, %10, proliferation features of these cells in vitro. Thus, %100) of HUVEC/HEp-2-CM for 130 hours to here we performed an in vitro direct and indirect 97 The Congress Is Supported By Turkish Airlines
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show the effect of concentration on the lower chambers as a chemoattractant of a control proliferation of HUVEC/HEp-2 cells by group. Then, cells (3.5 × 104) in 0.2 ml serum- xCELLigence RTCA DP (real-time cell analysis free DMEM medium were added to the upper dual purpose) system (Acea Biosciences, USA). chambers, cells (10.5 × 105) in 0.75 ml DMEM The cells were seeded in wells of E-Plates 16 and medium supplemented with 10% FBS was added proliferation was continuously monitored. The to the lower chambers as a chemoattractant of a xCELLigence RTCA DP instrument uses treated group. After incubation for 24 h, cells on noninvasive electrical impedance monitoring the upper membrane surface were wiped off, and technique to quantify cell proliferation, the cells that invaded across the matrigel morphology change, and attachmentn property- membrane were washed with phosphate-buffered label-free and real-time- by gold microelectrode saline (PBS), fixed with 100% methanol, rinsed biosensors in each well. Adherent cell in PBS, and stained with Giemsa staining. The proliferation rate is registered as increase in Cell invaded cells were captuerd under a light Index (CI). All experiments were performed at microscope (at 10x magnification). All invaded least in triplicate. cells counted by image J cell counter program. For direct co-cultivation system, the migration The filters used for migration assays were not and invasion assays were performed using coated with matrigel. All experiments were transwell chambers with 8 µm pore filters. The performed at least in triplicate. filters used for invasion assays were coated with Results: 50 µl pre-diluted matrigel (diluted at a ratio of In xCELLigence RTCA system, the impedance 1:10 with serum-free DMEM medium). Matrigel value was monitored for duration of 130 h and concentrations were optimized based on the expressed as a CI value (Figure 1 ve 2). migration capability of each cell line and According to the results of xCELLigence system, according to a previous in vitro assays (data not the HEp-2 cells co-cultured with %10 HUVEC- shown). Cells (3.5 × 104) in 0.2 ml serum-free CM (CI:6.553, p<0.05) proliferated slowly in DMEM medium were added to the upper comparison with control HEp-2 cells (CI: 7.299). chambers while 0.75 ml DMEM medium Moreover, HEp-2 cells co-cultured with %100 supplemented with 20% FBS was added to the HUVEC-CM (CI:3.168, p< 0.0001) proliferated
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much slowler than control HEp-2 cells (Figure in transwell chamber did not show more invasion 3A). The HUVEC cells co-cultured with %10 capacity than HEp-2 cells alone (%33.24, n.s., HEp-2-CM (CI:8.342, n.s., p>0.05) proliferated p>0.05 Figure 4A). By contrast, HUVEC co- less than control HUVEC cells (CI: 8.411). Also, cultured with HEp-2 (%25.64) in transwell HUVEC cells co-cultured with %100 HEp2-CM chamber were observed invading compared with (CI: 7.883 p<0.05) proliferated slower than the HUVEC cells alone (%14.80, n.s., p>0.05 control HUVEC cells (Figure 3B). Figure 4B). According to the results of invassion assays, the
HEp-2 cells co-cultured with HUVEC (%16.98)
Figure 1. Cell proliferation curves of HEp-2 cell line in different mediums as recorded by xCELLigence RTCA over 120 h.
Figure 2. Cell proliferation curves of HUVEC cell lines in different mediums as recorded by xCELLigence RTCA over 120 h. 99 The Congress Is Supported By Turkish Airlines
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Figure 3. The influence of conditioned mediums (10% and 100%) on proliferation of HEp-2 (A) and HUVEC cells (B) as stated by CI values.
Figure 4. Invasiveness in HEp-2 co-culture with HUVEC (A) and HUVEC co-culture with HEp-2 (B) cells lines. The data represents the percentage of invasiveness compared to the controls.
Statistical analysis: independent experiments. The significance of the Statistical analysis was conducted with differences was analyzed using Independent GraphPad version 6.0 software. Data are Student’s t tests. Differences were considered expressed as the mean ± SD from at least three significant at p < 0.05.
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Conclusion: References We showed that cancer cells and endothelial cells 1. Leemans CR, Braakhuis BJ, Brakenhoff have complex variety of communication with RH. The molecular biology of head and neck each other. We have seen that different co- cancer. Nat Rev Cancer 2011;11:9–22 culture systems may lead to different results. According to invasion assay, healthy cells (i.e. 2. Li J, Chen L, Qin Z. Multifaceted tumor HUVEC cells) limited the malignant activities of stromal fibroblasts. Cancer Microenviron HEp-2 cells by suppressing invasion. On the 2012; 5:187-193. other hand, cancer cells (i.e. Hep-2 cells) 3. Bhowmick NA, Neilson EG, Moses HL. increased the malignant activities of HUVEC Stromal fibroblasts in cancer initiation and cells by promoting invasion. progression. Nature 2004; 432:332-337. This study is a good indication that by using the 4. Lin EY, Pollard JW. Role of infiltrated xCELLigence RTCA system, in vitro leucocytes in tumour growth and spread. Br J understanding of the proliferation characteristics Cancer. 2004; 90:2053-2058. of cells in co-culture model can be enhanced. The microenvironment provided with HUVEC-CM 5. Ding ZY, Zou XL, Wei YQ. Cancer reduced the proliferation of cancer cells. There is microenvironment and cancer vaccine. also an inverse relationship between the Cancer Microenviron 2012; 5:333-344. proliferation rate of cancer cells and the 6. Zeng, Q., et al., Crosstalk between tumor and increasing concentration of CM. However, the endothelial cells promotes tumor microenvironment provided by CM from cancer angiogenesis by MAPK activation of Notch cells did not significantly affect the proliferation signaling. Cancer cell, 2005. 8(1): p. 13-23. of healthy cells.
Our study sheds light on the importance of microenvironment effect in cancer because the reaction of cells in mono-culture is different than co-culture and even tri-culture models.
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P.2. THE ACCURACY OF MICROSOFT KINECT IN UPPER EXTREMITY MOTION ANALYSIS
Mert Asım Karaoğlu1, Çağatay Kırmızıay1, Mert Canatan1, Özer Çevikaslan2, Gonca Şahiner3, Şermin Tükel4, Mehmet Türkan1
1 İzmir University of Economics, Department of Electrical and Electronics Engineering 2 İzmir University of Economics, Department of Software Engineering 3 İzmir University of Economics, Faculty of Health Sciences 4 İzmir University of Economics, Faculty of Medicine
Objectives: Movement analysis has important individuals and accuracy of the joint positions implications for neurological and aging studies were analysed during static posture of 90° and their clinical applications for early diagnosis, shoulder flexion. Second, movement trajectory following up the prognosis and for the was analysed from one person during diagonal assessment of the efficacy of therapeutic and shoulder movement. pharmacological interventions. Here we report Results: The system could record at the preliminary findings of a study, which approximately 27 fps. Accuracy of the ultimately aims at developing clinically valid, measurement of shoulder flexion was 97.42%. affordable and user-friendly movement analysis Movement trajectory analysis was successful method by using a markerless system. unless for high frequency jerky movements (see Methods: Microsoft Kinect v2 sensor was Figure 1). chosen to capture movement data. Kinect SDK Conclusions: We found Kinect v2 as a 2.0 and OpenCV library was used for data promising tool that requires further acquisition and Matlab environment was used for enhancements. In future studies, with more post processing and data analysis. First, upper advanced memory management techniques, extremity joint locations were captured on three more stable fps rate could be achieved.
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P.3. INTERACTION AND ACTIVE REGION DETERMINATION BETWEEN BACTERIAL CELLULOSE AND VARIOUS CELLULASE ENZYMES
A. Demet Demirag 1, Sefa Celik 2, Aysen E. Ozel1, Sevim Akyuz3
1 Istanbul University, Faculty of Science, Department of Physics, Vezneciler, 34134, Istanbul. 2 Istanbul University-Cerrahpasa, Faculty of Engineering, Department of Electrical and Electronic Engineering, 34320-Avcılar, İstanbul, Turkey. 3 Istanbul Kültür University, Faculty of Science and Letters, Department of Physics, 34156, Bakırköy- Istanbul, Turkey.
Cellulose is a renewable biopolymer which is the process, to ensure the digestion of strong solid most abundant in nature, formed by binding of molecules, such as cellulose, with maximum glucose units with β-1,4 glycosidic bonds. It's efficiency. Cellulases are classified according to one of the most important properties is forming a their mechanisms of action. In this study, the crystal structure that is different from other determination of the interaction mechanism of polysaccharides. Cellulose is divided into two Cellulose II, which is a bacterial cellulose, with groups as bacterial cellulose and vegetable cellulase enzymes by molecular docking method cellulose. Bacterial cellulose is a polymer whose based on key-lock theory is aimed. For this water retention capacity is higher than vegetable purpose, the most stable molecular geometry of cellulose, retains its shape and is suitable for the Cellulose II molecule was determined by modification during production. The main Density Functional Theory using Gaussian 05 application areas are serum lipids- and program. The scaled vibration frequencies of cholesterol-lowering effects, preventing the optimized geometry were calculated by using sudden rise of glucose level in urine effect, Molvib program. Molecular interactions between wound healing effect, protection against colon cellulose II with endoglucanase-Cellulose II, cancer, cardiovascular diseases, coronary Exoglucosidase-Cellulose II and β-glucosidase- thrombolysis, and production of surgical yarn. It Cellulose II have been determined. is also a very difficult industrial and molecular
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P.4. EVALUATION OF THYROID HORMONE PARAMETERS AND DYSLIPIDEMIA IN TYPE 2 DIABETIC PATIENTS
OMEED AKBAR ALI1, ALI HAMEED MAHMOOD1, SUZAN TABUR2
1 University of Gaziantep, Biochemistry Science and Technology, Gaziantep, Turkey. 2 University of Gaziantep, Internal Medicine, Gaziantep, Turkey.
(p<0.05), besides it has been found a significant Objectives: Thyroid diseases and diabetes are relationship among FT3, T-Cholesterol, and the most common endocrine disorders in clinical HDL-C. Also, it has been found a significant practice and have been shown to frequently affect relationship between FT4 and LDL-C (p<0.05) in each other. In this study, the aim was to assess the Anti-TPO (Negative) group. It didn't show a the relationship between Thyroid hormones and significant relationship between Thyroids lipid parameters in type II diabetic patients. parameters and lipid profiles (p>0.05) in the Methods: The thyroid parameters measured in Anti-TPO (Positive) group. our study were obtained using a Beckman Conclusions: This finding is accordant with Coulter Unicel DxI800, USA instrument; FBG, other studies showing that type 2 diabetes ALT, and lipid profiles were studied with increases thyroid disease and dyslipidemia risk. Beckman Coulter AU5800, US device and Findings from this study show that there may be HbA1c Biorad Variant 2, USA. a relationship between dyslipidemia and thyroid Results: It was found a significant increase in parameters, but advanced molecular tests are FBG, HbA1C, ALT, T-Cholesterol, HDL-C, needed to confirm it. In addition, we can foresee LDL-C, CRP, TSH parameters, (p<0.05), in type those thyroid hormone parameters and lipid II diabetic patients, whereas no significant profile can be used for in follow-up of Type 2 difference regarding FT4 and HGB parameters diabetes patients. were found. It has been found in the T2DM group Key words: Thyroid Diseases; Dyslipidemia; a significant relationship between TSH and TG Diabetes; Thyroid Hormones.
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P.5. CHANGES AT THE SUBCELLULAR LOCALIZATION AND DYNAMICS OF ANTI- APOPTOTIC p53, p57 AND Bcl-2 UNDER THE EFFECT OF COLLAGEN
Havva Tuyluoglu1, Ahmet Can Toksoz1, Hatice Isan1, Ranan Gulhan Aktas1
1 Cancer and Stem Cell Research Center, School of Medicine, Maltepe University, Istanbul, TURKEY.
OBJECTIVES: Collagen is considered as one Staining density was measured by using Zen of the most useful biomaterials. It is being used analysis system. in many medical fields including bioengineering RESULTS: P53 localization was clearly seen studies. Recent studies show that collagen might ER of the cells in un-coated group. Diffuse induce apoptosis. The mechanism of this staining of cytoplasm and staining in the nuclei induction is unclear. The tumor suppressor p53, of several cells was also observed. Under the the proto-oncogene Bcl-2, and a multifunctional effect of collagen, staining in nucleoli and diffuse protein, p57 have been linked to cell death by cytoplasmic staining were evident. Labeling apoptosis. Subcellular distribution of those patterns for both P57 and Bcl-2 was similar in proteins might change and this highly dynamic different groups. process is critical for cell fate determination. The CONCLUSIONS: Similar correlation was not current study was designed to examine the found between p57 and Bcl-2 subcellular changes at location of those three expressions. Subcellular localization of p53 proteins in the cells under the effect of collagen. shifted to nucleoli of the cells. Nuclear METHODS: HepG2 cells, a promising cell line accumulation of p53 plays a role in for bioartificial liver, were grown on either Type tumorigenesis and enhances the progress of I collagen coated or un-coated coverslips. malignant transformation. During the biomedical Samples were labeled with p53, p57 and Bcl2; studies; it should be noted that collagen might and examined under confocal microscope. induce both apoptosis and malign transformation of the cells.
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P.6. THE ASSOCIATION OF Β-ARRESTIN GENE POLYMORPHISM WITH ACROMEGALY DISEASE
Ayse Seda Akdemir1, Derya Metin Armağan1, Özge Polat Korkmaz2, Hande Mefkure Özkaya2, Pınar Kadıoğlu2, Nurperi Gazioglu3, Melek Öztürk1
1 Department of Medical Biology, Cerrahpaşa Medicine Faculty, Istanbul University-Cerrahpasa. 2 Department of Endocrinology and Metabolism, Cerrahpaşa Medicine Faculty, Istanbul University- Cerrahpasa. 3 Department of Neurosurgery, Medicine Faculty, Bilim University, İstanbul, Turkey.
Objectives: Acromegaly disesase occur as a Results: The genotype distributions of β- result of excessive growth hormone (GH) caused arrestin-2 were found as CC:68,2%; CT:28,2%; by the pituitary adenomas (PA). Some of the TT:3,6% in AP and treated acromegaly patients (AP) shows CC:60,6%;CT:32,3%;TT:7,1% in controls. In all resistance to somatostatin analogs treatment. β- AP, there was a significant correlation between arrestins are involved in somatostatin receptors macro/microadenoma percentages and CT+TT intracellular endocytic mechanisms and genotypes and T-allel (p=0,006). The genotype signaling. β-arrestins might be play a role in the distributions of AP with good response to development of PA and the response to medical medical treatment were compared with the treatment. We aimed to investigate genotype clinical data of the disease, a significant distribution of β-arrestin-2 SNP in Turkish relationship was found between CT+TT population, also the possible relationship genotypes and T allel and macro/microadenoma between the SNP and disease risk and medical percentages (respectively p=0,017;p=0,005) and treatment response in AP. postop-3.months GH levels (respectively Methods: Genotypes of rs34230287 in the p=0,011;p<0,001). In AP with poor response to promoter region of the β-arrestin-2 gene were medical treatment, the end-GH levels were found determined by RT-PCR method in 110 APs and higher with T-allel (p=0,025). 99 healthy controls. Conclusions: We showed that while there was no correlation between polymorphic genotypes 107 The Congress Is Supported By Turkish Airlines
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of β-arrestin-2 gene with acromegaly disease and medical treatment response groups and β- medical treatment response in Turkish arrestin-2 SNP. population, there was a relationship between the (This study was supported by İstanbul University clinical characteristics of the patients in the Scientific Research Projects;TP26147)
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P.7. EFFECTS OF MORIN AND SORBIC ACID ON THE ACTIVITY OF HUMAN ERYTHROCYTES GLUTATHIONE REDUCTASE
Uğur Güller
Igdir University, Faculty of Engineering, Department of Food Engineering, 76100-IĞDIR.
ABSTRACT INTRODUCTION Glutathione reductase (GR) is an important part Glutathione reductase (GR, EC 1.8.1.7) catalyses of the enzymatic antioxidant defence system, the reduction of glutathione disulphide (GSSG) catalyses electron transfers among disulphide to sulfhydryl form glutathione (GSH) in the substrates and reduced pyrimidine nucleotides presence of nicotinamide adenine dinucleotide (NADPH). The task of the GR enzyme is to phosphate (reduced form) (NADPH). preserve the oxidized glutathione (GSSG) and Glutathione, an important part of the non- reduced glutathione (GSH) balance (GSH/GSSG enzymatic defence system, is a tripeptide ratio). In the case of GR inhibition, that is, GSH consisting of glutamine, cysteine and glycine. imbalance, some pathological disorders may be Glutathione is present in the cell as reduced seen. (GSH) and oxidized (GSSG) (1). One of the most In this study, GR enzyme was purified in a single important targets of the reaction catalysed by GR step by using 2', 5' ADP-Sepharose 4B affinity is to maintain the GSH/GSSG ratio, a marker of chromatography from human erythrocytes. The cellular redox status, in the cell. This ratio is inhibitory effect of sorbic acid and morin which approximately 500/1 in erythrocyte cells. When are used in the food sector, was investigated on this ratio is lower, erythrocyte cells become GR activity obtained with 8.91 EU/mg protein haemolysis (2). Thiol groups of GSH are specific activity. Both chemicals inhibited GR potential reducing agents. As a major with the IC50 value of 2.01 mM and 1.13 mM antioxidant, GSH is involved in the respectively. detoxification of various electrophilic compounds and peroxides. In addition to detoxification, the GSH has roles in other cellular 109 The Congress Is Supported By Turkish Airlines
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reactions such as glyoxalase system, reduction of Preparation of the hemolyzate ribonucleotides to deoxyribonucleotides, Erythrocyte pellet was obtained from Atatürk thiosulfide exchange reactions and regulation of University, Medicinal Faculty Blood Centre. protein and gene expression (3). In case of GR Erythrocyte pellet haemolysed with 5 volume of inhibition, GSH deficiency causes oxidative ice-cold water and then centrifuged at 10 000g damage risk in cell and pathologies such as for 30 min (at 4°C,) to remove the residual intact cancer, neurodegenerative disorders, cystic cells and membranes (9). fibrosis, HIV and aging can be seen (4). Besides, Purification of GR high GSSG concentration inhibits many The hemolyzate was loaded onto the 2’, 5’-ADP important enzymatic pathways including protein Sepharose 4B affinity column previously synthesis (5,6). prepared and equilibrated with 0.1 M K- MATERIALS AND METHODS acetate/0.1 M K-phosphate buffer (pH 6.0) Then, Materials column was washed with 25 mL of 0.1 M K- 2’, 5’-ADP Sepharose 4B was purchased from acetate + 0.1 M K-phosphate, pH 6.0 and 25 mL Pharmacia. NADPH, GSSG, the protein assay of 0.1 M K-acetate+ 0.1 M K-phosphate, pH reagent and the chemicals for electrophoresis 7.85. Washing continued with 50 mM of K- were purchased from Sigma Chem. Co. All other phosphate including 1 mM EDTA (pH 7.0) until chemicals used were analytical grade and the absorbance difference between third washing purchased from either Merck or Sigma. buffer and recovered solution from bottom were Measurements of GR activity became 0.05 at 280 nm. GR was eluted with a Activity assay of GR was performed with 1-ml gradient of 0-0.5 mM GSH and 0-1m M NADPH cuvettes at 340 nm spectrophotometrically as in 50 mM K-phosphate, containing 1 mM EDTA described in Carlberg and Mannervik (7). (pH 7.0). Active fractions were collected and Protein determination dialyzed with 50 mM of K-phosphate including Quantitative protein determination was measured 1 mM EDTA (pH 7.0) at 4 °C. spectrophotometrically at 595 nm according to In Vitro Inhibition Studies the Bradford method (8). Inhibitory effects of sorbic acid and morin were examined on enzyme activity. On this purpose,
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enzyme activities in varying concentrations of formation is essential in the metabolism of GSH- inhibitors were measured and half maximal dependent defence reactions. Therefore, the role inhibitory concentrations (IC50) were calculated of GR in enzymatic defence is extremely by drawing “activity%-[I]” graphs. important. The investigation of GR inhibitors is RESULTS AND DISCUSSIONS becoming the focus of attention for both the Reactive oxygen species (ROS) are inevitable antimalarial and anticancer activities and for the products of normal metabolism. However, the elucidation of the effects of them on the formation of an excess of ROS, including antioxidant system.
–• peroxide (H2O2) and superoxide anions (O2 ), is In this study the purification of the erythrocyte toxic to the cell. The organisms have antioxidant GR was performed by the single chromatography defence systems to control the effects of ROS step method by means of 2',5'-ADP Sepharose- and other radicals to the cell. GSH is an important 4B affinity gel chromatography. The enzyme was part of the non-enzymatic antioxidant system and purified 2173.7-fold with a specific activity of is generated by the reduction of GSSG by a 8.91 EU/mg protein and overall yield of 44.8% reaction with the enzyme GR where NADPH is (Table 1). the electron donor. The recycle pathway for GSH
Table 1. Purification steps of glutathione reductase from human erythrocytes Sample Type Total Total Total Specific Activity Protein Yield Purification Volume Protein Activity Activity (EU/ml) (mg/ml) % Coefficient (ml) (mg) (EU) (EU/mg) Hemolyzate 20 0.085 20.8 416 1.7 0.0041 100 1 Affinity 3 0.254 0.0285 0.086 0.762 8.91 44.8 2173.7 Chromatography
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The process was performed in a very short time, and our results are better when compare to the previous studies (10,11). The inhibitory effects of sorbic acid and morin on the activity of human erythrocyte GR were also examined. The inhibitor concentration which reduced the enzyme activity by half was calculated from % activity-[inhibitor] graphs (Figure 1). It was Figure 1. Activity%-[inhibitor] graphs for (a) observed that both chemicals inhibited GR with sorbic acid and (b) morin chemicals. the IC50 value of 2.01 mM and 1.13 mM respectively. The GR enzyme has been purified In conclusion, either such molecules can be used many times from human erythrocytes and many as pioneer for discovery of novel effective GR studies have been carried out on its inhibition. inhibitors for medicinal chemistry applications For example, in vitro inhibition of some (anti-malaria, anti-cancer, etc.) or attention will nitroaromatic compounds and some commonly be paid to the use of these chemicals for healthy used drugs on GR was investigated (10,11,12). individuals.
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References 7. Carlberg, I. and Mannervik B., 1975. J. 1. Griffith, O.W., 1999. Free Radic. Biol. Biol. Chem. 250, 5475-5480. Med., 27: 922–935. 8. Bradford, M.M., 1976. Anal. Biochem., 2. Keha E., Küfrevioğlu Ö.İ., 2010. Aktif 72, 248. Press, Erzurum, 653s 9. Beutler, E., 1984. Inc. Orlando, FL 3. Mullineaux, P., Creissen G.P., 1997. 32887, London. Cold Spring Harbor Laboratory Press. 10. Akkemik, E., Şentürk, M., Özgeriş, F.B., 4. Townsend, D.M., Kenneth D., Tew K.D., Taşer, P., Çiftci, M., 2011. Turk J Med Tapiero H., 2003. Biomedicine & Sci, 41 (2): 235-241. Pharmacotherapy 57, 145-155. 11. Karaman, M., Akkemik, E., Budak, H., 5. Vijayalingam, S., Parthiban A., Ciftci, M. 2012, J Enzyme Inhib Med Shanmugasundaram K.R., Mohan V., Chem., 27(1): 18–23. 1996. Diabet. Med.,13: 715–719. 12. Çakmak, R., Durdagi, S., Ekinci, D., 6. Akerlund, B., Tynell E., Bratt G., Şentürk, M., Topal, G. 2011, Bioorg Med Bielenstein M., Lidman C., 1997. J. Chem Lett., 21: 5398–5402. Infect, 35:143–147.
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P.8. THE ASSOCIATION OF miR-101-3P AND ADAM15 GENE EXPRESSIONS IN PROSTATE CANCER
Rozhgar A. KHAILANY1,2, Muhammad SAFDAR3, Yasmin JUNIJO3, Naser GILANI2, Khandakar A S M SAADAT2 and Belan O. KANABE4
1Department of Biology, College of Science, University of Salahaddin, Erbil, Iraq. 2Department of Medical Biology and Genetics, University of Gaziantep, Gaziantep, Turkey. 3Department of Molecular Biology, Faculty of Science and Technology, Government Virtual University of Pakistan, Lahore, Pakistan; 4Department of Biology, Gaziantep University, Gaziantep, Turkey.
Background: The most common malignancy in miR-101-3p that selectively inhibits ADAM15 men is prostate cancer next to lung cancer. Drug expression and increase activity of resistance in the treatment of cancer still remains enzalutamide. MiR-101-3p and ADAM15 a major clinical concern. Resistance to expressions were measured in prostate cancer enzalutamide is seen in 60-75% of the cell lines (LNCaP, PC-3, Du145, and recurrences in prostate cancer. The anti- CA7T2CL) and in PNT2 (control) by Real-time testosterone enzalutamide may gain agonistic PCR. The expression levels of miR-101-3p were property by suppressing androgen-receptors significantly decreased while the expression (AR). ErbB (HER2-HER3) receptors are known level of ADAM15 was increased in LNCaP cell cleaved by an ectodomain sheddase, ADAM15, line according to PNT2 cell line. to liberate HER2-HER3 heterodimer domain in In brief, we suggest that the inhibition of most metastatic cases. So, enzalutamide cannot ADAM15 by miR-101-3p specifically may inactivate AR receptors by binding ectodomain provide a therapeutic approach by sensitizing portion of it. Recently, microRNA-101’s role in ErbB+ breast cancer cells to enzalutamide prostate cancer has been demonstrated but targeting ECD of HER2-HER3 heterodimer. mechanism is not fully understood yet. Our aim Keywords: miR-101-3p, ADAM15, Prostate is to suppress ectodomain sheddase activity with cancer, qRT-PCR
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P.9. EXPRESSION ASSOCIATION OF MiR-103 WITH RENAL CELL CARCINOMA
Rozhgar A. KHAILANY1,2, Mustafa S. AL-ATTAR1, Naser GILANI2, Belan O. KANABE3, Khandakar A S M SAADAT2
1Department of Biology, College of Science, University of Salahaddin, Erbil, Iraq. 2Department of Medical Biology and Genetics, University of Gaziantep, Gaziantep, Turkey. 3Department of Biology, Gaziantep University, Gaziantep, Turkey.
Renal cell carcinoma is the third most common samples were grouped according to the types of urologic cancer and the seventh most common renal carcinoma and clinical characteristics of malignancy overall. MicroRNAs (miRNAs) patients, including gender and average age. initiate translational controlling and play a Expression level of MiR-103 was significantly critical role in developmental timing. Presently, increased (up-regulated) in renal cell carcinoma it is determined miRNAs play an important roles tissues compared to normal tissues. in several physiological and pathological Consequently, the MiR-103 could be potentially conditions, particularly deregulation in various used as diagnostic markers in renal cell type of cancers. The goal of this study was to carcinoma. However, further studies are evaluate the expression level of MiR-103 in renal mandatory to a better understanding and cell carcinoma. MicroRNAs expression level of confirmation of our preliminary findings. 40 paired tumor and normal tissue samples were Key words: Renal cell carcinoma, Expression measured by quantitative real time polymerase analysis, MicroRNAs, qRT-PCR. chain reaction (qRT-PCR) technique. The
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AUTO INDEX
A-F
A. Demet Demirag Elif Pala
Ahmet Can Toksoz Emine Yavuz
Ahmet Ulu Ensar Erel
Alı Hameed Mahmood Ezgi Kıyga
Ali Noma Fatma Gozde Yuce
Ayfer Pazarbaşı Derya Metin Armağan
Aysen E. Ozel Devrim Pesen Okvur
Ayse Seda Akdemir
Barbaros Şahin Karagün G-K
Belan O. Kanabe Gamze Comertpay
Belisa Kaleci Pilafi Gamze Gungor
Berna Kaya Uğur Gokcen Guvenc
Betül Kocamer Şimşek Gokhan Gokaslan
Burçin Altınbaş Gonca Şahiner
Burhan Ateş Göktürk Maralcan Çağatay Kırmızıay Gülper Nacarkahya Canbolat Gürses Gülsevinc Ay Ebru Kuyumcu Savan
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Gülşah Erel Mehmet Çiftci
Halide Sedef Karaman Mehmet Ozaslan
Hasan Akbaba Mehmet Solakhan
Hatice Duran Mehmet Türkan
Hatice Isan Mehmet Yılmaz
Hande Mefkure Özkaya Melek Öztürk
Havva Tuyluoglu Mert Asım Karaoğlu
Hülya Çiçek Mert Canatan
Işıl Öcal Merve Gökşin Karaaslan
I. Nur Uslu Meryem Kara
İbrahim Ozaslan Muhammed Karaman
İmren Özcan Muhammad Safdar
İsmet Yilmaz Muhammad Safdar
Khandakar A S M Saadat Murat Yalcin
Kutay Içöz Mustafa S. Al-Attar
Mustafa Kotmakçı
L-R Mustafa Yıldırım
Lütfiye Özpak Naser Gilani
M. Koyutürk Nasir Niaz
Mecit Özdemir Necla Benlier
Mehmet Bertan Yılmaz Nevhiz Gündoğdu
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Nevin Uysal Saliha Dinç
Neytullah Turan Seda N. Ilgaz
Nilgun Tanriverdi Sefa Celik
Nurperi Gazioglu Selim Buyukkurt
Osman Demirhan Serdar Akaltun
Omeed Akbar Alı Seval Kul
Öner Dikensoy Sevgi Balcıoğlu
Özer Çevikaslan Sevim Akyuz
Özge Polat Korkmaz Suzan Tabur
Öznur Güngör Suleyman Ganıdaglı
Özlem Altindağ Süleyman Köytepe
Özlem Nuray Sever Şermin Tükel
Pınar Güller Taner Arpacı
Pınar Kadıoğlu Tekin Izgi
Pınar Tumturk Tuba Denkçeken
Ranan Gulhan Aktas Tülin Cora
Rozghar A. Khailany Türkan Turgay
Uğur Güller
S-Z Ümit Luleyap
Sabriye Kocaturk-Sel Ünal Akar
Samir Abbas Veli Serkan Kolat
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Yasmin Junijo Zeynep Betül Sari
Yasemin Yıldızhan Zelal Adıguzel
Yücel Başpınar Zeliha Yıldırım
Yüksel Çetin
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