The UKRAINIANBIOCheMICALJOURNALVolume 88, Special Issue, 2016

NATIONAL ACAdeMy Of SCIeNCeS Of UKRAINePALLAdIN INSTITUTe Of BIOCheMISTRy

Bridges in Life Sciences 11th Annual

Scientific Conference Prague, Czech

Republic

April 7 – 10, 2016

Table of Contents

Organizing Committee 3

Agenda of the Bridges in Life Sciences 10th Conference 4

RECOOP HST Association 17

Croatian Medical Journal 20

Abstracts 21

Common Mechanism of Diseases 22

Cardiovascular Research in Progress 31

Nanomedicine Research 37

RECOOP Research in Progress 56

Infections and Inflammation 64

RECOOP Research in Progress 70

Experimental Works of RECOOP Members (Ukraine) 78

RECOOP Visegrad Scholarship Program 114

Conference Participants 116

Organizing Committee USA - Cedars-Sinai Medical Center, Los Angeles, California Edward Prunchunas Senior Vice President for Finance and Chief Financial Officer, Cedars-Sinai Medical Center & Chairman of the Supervisory Board of the RECOOP HST Association Shlomo Melmed, M.D. Senior Vice President, Academic Affairs, Dean of the Medical Faculty, Cedars-Sinai Medical Center Sandor G. Vari, MD Director, International Research and Innovation Management Program, Cedars-Sinai Medical Center & President of the RECOOP HST Association RECOOP HST Association Veronika Puska, Grant and Project Manager, RECOOP HST Association Czech Republic Jan Pitha, Head of Laboratory for Atherosclerosis Research, Centre for Experimental Research, Institute for Clinical and Experimental Medicine (IKEM) Editor: Sandor G. Vari, M.D.

Agenda

April 8, 2016 (Friday) Congress Hall

8:30 – 9:00 Message from the President of RECOOP

Sandor G. Vari, Director, International Research and Innovation Management Program, Cedars-Sinai Medical Center, Los Angeles, CA, USA & President of RECOOP HST Association

Message from the 35th President of the United States on Friday, January 20, 1961 “My fellow Americans, ask not what your country can do for you, ask what you can do for your country.” Message from the President of RECOOP HST Association on April 8, 2016 My fellow RECOOP members, ask not what RECOOP can do for you, ask what you can do for RECOOP. 9:00 – Keynote lecture: 9:00 – 9:15 RECOOP Project Common Mechanism of Diseases

Diet induced obesity and impaired glucose metabolism in elderly rats treated with Liraglutide (Victoza) and Metformin. Robert Gaspar, Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, Hungary

9:15 – 9:25 Young Scietists Contribution – Electronic Interactive Poster Presentation Five-minute power point presentation

Glucose and insulin tolerance test in obese, elderly rats with impaired glucose metabolism treated with Liraglutide (Victoza) and Metformin Kalman Szucs, Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, Hungary

Plenary Session: Common Mechanism of Disease Oral presentation is max 15 minutes with all tolerance after 15 minutes no tolerance. Electronic Interactive Poster Presentation EIPP = 5-minute power point presentation and 5-minute discussion. Posters in Conference Room 5 Chairs: Ines Drenjančević, Josip Juraj Strossmayer University of Osijek, Croatia Jan Pitha, IKEM, Prague, Czech Republic Elizabeta Has-Schön, Josip Juraj Strossmayer University of Osijek, Croatia 9:25 – 9:40 Alteration of inflammatory markers and SSAO activity in diet induced diabetic, obese elderly rats treated with Liraglutide (Victoza) and Metformin. Tamás Tábi Department of Pharmacodynamics, Faculty of Pharmacy, Semmelweis University, Budapest, Hungary

9:40 – 9:55 Changes in leptin, adiponectin, insulin and gonadal steroid receptors’ response in the brain of diet induced diabetic, obese elderly rats’ brain treated with Liraglutide (Victoza) and Metformin. Marija Heffer & Senka Blazetic Department of Medical Biology, School of Medicine, Josip Juraj Strossmayer University of Osijek, Croatia 9:55 – 10:00 Degenerative changes in the brain of diet induce diabetic, obese elderly rats treated with Liraglutide (Victoza) and Metformin. Srećko Gajović Professor of Histology and Embryology, University of Zagreb School of Medicine, Croatia 10:00 – 10:15 Markers of Blood Oxidative Stress and Antioxidative Enzymes Activity in Obese Diabetic Elderly Rats Treated with Metformin or Liraglutide Ines Drenjančević Department of Physiology and Immunology, Faculty of Medicine, University J. J. Strossmayer Osijek, Croatia 10:15 – 10:30 Fatty acid changes in plasma, visceral and subcutaneous adipose tissue in diabetic, obese elderly rats treated with Liraglutide (Victoza) and Metformin. Eva Nagy Doctorate School of Animal Husbandry, Faculty of Agriculture and Nutritional Sciences, University of Debrecen, Hungary 10:30 – 10:45 Renal pathology alterations in diet induced diabetic, obese elderly rats treated with Liraglutide(Victoza) and Metformin. Peter Boor Institute of Pathology and Department of Nephrology, RWTH University Aachen, Germany 10:45 – 11:00 Changes in leptin and adiponectin receptors and myocytes’ contractility of uterus in diabetic, obese elderly rats treated with Liraglutide (Victoza) and Metformin. Robert Gaspar, Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, Hungary Bone density change in diabetic, obese elderly rats treated with Liraglutide (Victoza) and Metformin. (only read in the abstract book) Zora Krivosikova, Department of Clinical and Experimental Pharmacotherapy, Slovak Medical University, Medical Faculty, Bratislava, Slovak Republic 11:00 – 11:30 Coffee Break Conference Room 3

Arts and Sciences Exhibits’ Opening Ceremony Sandor G. Vari, Cedars – Sinai Medical Center, Los Angeles, CA, USA Julia Mallasz, Semmelweis Publishing & Multimedia Studio, Budapest, Hungary

11:30 – 11:45 Panel discussion Common Mechanism of Disease

Moderators: Robert Gaspar, Faculty of Pharmacy, University of Szeged, Hungary Tamás Tábi, Semmelweis University, Budapest, Hungary Sandor G. Vari, Cedars – Sinai Medical Center, Los Angeles, CA, USA

11:45 – 13:00 Cardiovascular Research in Progress Chairs: Eva Szoko, Semmelweis University, Budapest, Hungary Oksana Zayachkivska, Danylo Halytsky Lviv National Medical University, Ukraine Sandor G. Vari, Cedars – Sinai Medical Center, Los Angeles, CA, USA Oral Presentation 11:45 – 12:00 Cardiovascular risk factors and their impact on vascular parameters in women are modified by reproductive status and smoking, longitudinal study. Jan Pitha, Center for Experimental Medicine, Institute for Clinical and Experimental Medicine, Prague, Czech Republic 12:00 – 12:15 Shared decision making in life style and nutrition for intervention in woman with risk factors in cardiovascular health Slavica Juric Petricevic Department of Emergency Medicine in Split, Department of Family Medicine, University of Split School of Medicine, Split, Croatia Electronic Interactive Poster Presentation Posters in Conference Room 5 12:15– 12:25 Influence of acute and intermittent hyperbaric oxygenation on the reactivity of the blood vessels in healthy Sprague-Dawley rats Zrinka Mihaljevic Department of Physiology and Immunology, Faculty of Medicine Josip Juraj Strossmayer,

University of Osijek, Croatia.

12:25 – 12:35 Comprehensive analysis of ischemic conditioning techniques in a porcine model of acute myocardial infarction Andras Makkos Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary 12:35 – 12:45 The Role of Oxidative Stress in Development of Impaired Vascular Response Under the Influence of High Salt Intake in Sprague-Dawley Rats Anita Cosic Department of Physiology and Immunology, Faculty of Medicine University of J.J. Strossmayer in Osijek, Osijek, Croatia 12:45 – 13:00 Evaluation of putative cannabinoid ligands as modulators of neurodegeneration Yulia Senkiv Department of Regulation Cell Proliferation and Apoptosis, Institute of Cell Biology NAS of Ukraine, Lviv, Ukraine 13:00 – 14:00 Lunch Break

14:00 – 16:00 Nanobiomedicine Research Chairs: Rostyslav Stoika, Institute of Cell Biology, NASU, Lviv, Ukraine Tatiana Borisova, Palladin Institute of Biochemistry NASU, Kyiv, Ukraine Artur Podhorodecki, Wroclaw University of Technology, Poland Wroclaw Srecko Gajovic, University of Zagreb School of Medicine, Croatia Oral presentations 14:00 – 14:15 Research activity at Institute of Cell Biology, NAS of Ukraine: achievements, problems and perspectives of development within RECOOP-HST Association. Rostyslav R. Stoika Department of Regulation of Cell Proliferation and Apoptosis, Institute of Cell Biology (ICB), NAS of Ukraine, Lviv, Ukraine 14:15 – 14:30 Synthesis and surface functionalization of inorganic nanocrystals for their biomedical applications Artus Podhorodecki Department of Experimental Physics, Wroclaw University of Technology, Wrocław, Poland 14:30 – 14:45 Biocompatibility and stem cell labelling properties of maghemite nanoparticles Srecko Gajovic Croatian Institute for Brain Research, University of Zagreb School of Medicine, Zagreb, Croatia 14:45 – 15:00 S, N - containing carbon dots: fluorescent properties and application in cells experiments Mariia Dekaliuk Lab. of Nanobiotechnologies, Palladin Institute of Biochemistry of the NAS of Ukraine, Kiev EIPP – Electronic Interactive Poster Presentations Posters in Conference Room 5 15:00 – 15:10 Neurotoxic properties of carbon dots synthesized from sulphur-containing carbohydrate precursor Maryna Dudarenko Department of Neurochemistry and Lab.of Nanobiotechnology, Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine 15:00 – 15:20 CdSe/CdS quantum dots functionalization by d-penicillamine Anna Lesiak Department of Experimental Physics, Wroclaw University of Technology, Wrocław, Poland 15:20 – 15:30 Synthesis of Cd-based core shell nanocrystals for bio-imaging Maciej Chrzanowski Department of Experimental Physics, Wroclaw University of Technology, Wrocław, Poland

15:30 – 16:00 Panel Discussion: Future of RECOOP Nanobiomedicine Research Moderators: Rostyslav Stoika, Institute of Cell Biology, NASU, Lviv, Ukraine Tatiana Borisova, Palladin Institute of Biochemistry NASU, Kyiv, Ukraine Artur Podhorodecki, Wroclaw University of Technology, Poland Wroclaw Srecko Gajovic, University of Zagreb School of Medicine, Croatia

16:00 – 17:00 Coffee Break Arts and Sciences Exhibit Conference Room 3 RECOOP Play Room Conference Room 2

Each artist – scientist has to introduce the artwork presented at the Conference 17:00 – 19:30 RECOOP Research in Progress – Young Scientists Forum Chairs Robert Gaspar, Faculty of Pharmacy, University of Szeged, Hungary Tamás Tábi, Semmelweis University, Budapest, Hungary Marija Heffer, Josip Juraj Strossmayer University of Osijek, Croatia Tibor Ertl, University of Pecs, Hungary Oral presentation 17:00 – 17:15 Sex-specific oxidative-antioxidative status in the liver of the chronically and acutely stressed rats Elizabeta Has-Schön for Rosemary Vuković Department of Biology, University J. J. Strossmayer of Osijek, Osijek, Croatia; 17:15 – 17:30 Search for New Antifibrotic Agents among (amino)iminothiazolidinones Danylo Kaminskyy Department of Pharmaceutical, Organic and Bioorganic Chemistry, Danylo Halytsky Lviv National Medical University, Ukraine; 17:30 – 17:40 Birth outcome yearly dustrubution at Department of Gynecology and Obstetrics, University Hospital Center Osijek Andrijana Müller Department of Gynecology and Obstetrics, University Hospital Center Osijek, Faculty of Medicine, J.J. Strossmayer University of Osijek, Croatia Young Scientists Forum EIPP – Electronic Interactive Poster Presentations Posters in Conference Room 5 17:40 – 17:50 Metformin may influence neurodegeneration and neuroinflammation in Sprague Dawley rats on high fat/sugar diet in a sex specific manner Marta Balog and Milorad Zjalic Faculty of Medicine, J.J. Strossmayer University of Osijek, Croatia 17:50 – 18:00 The importance of autophagy in the antiapoptotic effect of resveratrol

Fruzsina Bagamery Department of Pharmacodynamics, Faculty of Pharmacy, Semmelweis University, Budapest, Hungary 18:00 – 18:10 Application of THY1 – YFP neural stem cells in regenerative neuroscience Ivan Alic Department of Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia 18:10 – 18:20 ARHGAP25 Rac-GAP has an important role in autoantibody-induced model of rheumatoid arthritis Tim F. Svanya Department of Physiology, Faculty of Medicine, Semmelweis University, Budapest, Hungary 18:20 – 18:30 The use of formalin-fixed paraffin-embedded samples in early diagnosis of diseases Boglarka Donczo Horváth Csaba Laboratory of Bioseparation Sciences, University of Debrecen, Debrecen, Hungary 18:30 – 18:40 The role of gamma aminobutyric acid (GABA) in the regulation of endocervical epithelium secretory activity Marta Skelin Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Croatia 18:40 – 18:50 Analyses of impact circadian dysfunctions on the occurrence of disautotonia in medical student population Maryana Zvir Physiology Department, Lviv National Medical University, Lviv, Ukraine 18:50 – 19:00 Plasma membrane Ca2+-pump selective regulators – calix[4]arene C-90 and imidazo[1,2-a]azepine – change parameters of uterus smooth muscles contraction Iuliia Mazur Department of Muscle Biochemistry, Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv, Ukraine 19:00 – 19:10 Does endogen PACAP have any role in the development of retinopathy of prematurity? Timea Kvarik Departments of Anatomy, Pacap-Lendulet Research Group and Obstetrics and Gynecology, University of Pecs, Hungary 19:10 – 19:20 The comparison of hydrogen sulfide NSAID derivates in the modulation of gastro-esophageal integrity. Irena Pshyk-Titko Department of Physiology, Lviv National Medical University, Ukraine 20:00 – 22:00 Dinner

April 9, 2016 (Saturday) Congress Hall

8:30 – 9:00 Summary of the Conference’s Progress

Sandor G. Vari 9:00 – 11:00 Annual Review of the Research Activities in RECOOP Session I Chairs Srecko Gajovic, University of Zagreb School of Medicine, Croatia Sandor G. Vari, Cedars – Sinai Medical Center, Los Angeles, CA, USA Eva Szoko, Semmelweis University, Budapest, Hungary Session I

9:00 – 9:15 Antioxidants Selenomethionine and D-Pantethine Decrease Negative Side

Effects of Doxorubicin in Nk/Ly Lymphoma-Bearing Mice

Rostyslav R. Panchuk

Department of Regulation of Cell Proliferation and Apoptosis, Institute of Cell Biology

National Academy of Sciences Ukraine, Lviv, Ukraine

9:15 – 9:30 Ovariectomy and chronic stress lead toward leptin resistance in satiety centers and insulin resistance in hippocampus of Sprague-Dawley rat Vedrana Ivic Josip Juraj Strossmayer University of Osijek, Faculty of Medicine Osijek, Department of Medical Biology and Genetics, Osijek, Croatia 9:30 – 9:45 Diurnal variation in cholesterol 7α-hydroxylase activity is determined by the -203A>C polymorphism of the CYP7A1 gene Tereza Blahová Laboratory for Atherosclerosis Research, Institute for Clinical and Experimental Medicine, Czech Republic 9:45- 10:00 Putative anticancer potential of novel 4-thiazolidinone derivatives: toxicity towards rat C6 glioma cells and correlation of general toxicity with balance of free radical oxidation in rats Lesya Коbylinska Department of Biochemistry, Danylo Halytsky Lviv National Medical University, Ukraine 10:00 – 10:15 The effects of estrogen on the α2-adrenergic receptor subtypes in late pregnant uterine function in vitro Judit Hajagos-Toth Department of Pharmacodynamics and Biopharmacy, University of Szeged, Szeged, Hungary 10:15 – 10:30 H. pylori in sedentary males is linked to higher heart rate, sympathetic activity and insulin resistance but not inflammation or oxidative stress Mykhailo Pliatsko Department of Internal Medicine №1, Danylo Halytskyi Lviv National Medical University, Lviv, Ukraine

10:30 – 10:45 Morphological and pathological response in primary systemic therapy of patients with breast cancer and the prediction of disease free survival Gyöngyver Szentmartoni Semmelweis University, Department of Clinical Oncology, Budapest, Hungary 11:00 – 11:30 Coffee Break Arts and Sciences Exhibit Conference Room 3 RECOOP Play Room Conference Room 2

Each artist – scientist has to introduce the artwork presented 11:30 – 12:00 Panel Discussions Annual Review of the Research Activities in RECOOP

Moderators: Srecko Gajovic, University of Zagreb School of Medicine, Croatia Sandor G. Vari, Cedars – Sinai Medical Center, Los Angeles, CA, USA Eva Szoko, Semmelweis University, Budapest, Hungary

12:00 – 13:00 Young Scientist Forum - Infection and Inflammation Chairs Pavel Bostik, Faculty of Military Health Sciences, Univ. of Defence, Czech Republic Shubhada Bopegamage, Slovak Medical University, Bratislava, Slovakia 12:00 – 12:10 Induction of cytokines by CVB2-Ohio and its mutants Brigita Benkoova Enterovirus Laboratory, Faculty of Medicine, Slovak Medical University in Bratislava, Slovakia 12:10 – 12:20 Possible mechanisms involved in the development of drug resistance in Ruk/CIN85-overexpressing MCF-7 cells Ganna Pasichnyk Laboratory of Cell Signaling, Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kiev, Ukraine 12:20 – 12:30 QSAR Studies of 4-thiazolidinone derivatives showing antitrypanosomal activity Anna Kryshchyshyn Department of Pharmaceutical, Organic and Bioorganic Chemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine; 12:30 – 12:40 Plasmid-mediated fluoroquinolone resistance in Enterobacteriaceae Orsolya Szabó Institute of Microbiology, Semmelweis University, Budapest, Hungary 12:40 – 12:50 Selected concepts and investigations among thiazolo[4,5-b]pyridines as perspective anticancer and anti-inflammatory agents Andrii Lozynskyi Department of Pharmaceutical, Organic and Bioorganic Chemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine

13:00 – 14:00 Lunch Break 14:00 – 16:00 Annual Review of the Research Activities in RECOOP Session II Chairs Charles F. Simmons, Cedars – Sinai Medical Center, Los Angeles, CA, USA Iuliana Ceausu, “Carol Davila” University of Medicine and Pharmacy, Bucharest, Romania Peter Boor, RWTH University Aachen, Germany Session II

14:00 – 14:15 Relationship of soluble receptors for advanced glycation end products and metabolic syndrome in adolescents from Bratislava region Radana Gurecka Institute of Molecular Biomedicine, Faculty of Medicine and Institute of Medical Physics, Biophysics, Informatics and Telemedicine, Comenius University, Bratislava, Slovakia 14:15 – 14:30 The effects of anthocyanin-rich wheat diet on the oxidative status and behavior of rats Katarina Jansakova

Institute of Molecular Biomedicine, Faculty of Medicine, Comenius University, Bratislava, Slovakia 14:30 – 14:45 Interleukin 6/Wnt interactions in rheumatoid arthritis: Interleukin 6 inhibits Wnt signaling in synovial fibroblasts and osteoblasts Khrystyna Malysheva Institute of Cell Biology National Academy of Sciences Ukraine, Lviv, Ukraine

14:45 – 15:00 Exploiting Hydrogen Sulfide of Novel 4-Thiazolidinone Derivatives in Cytoprotection of Small Intestine Under Indometacin-Induced Injury Iryna Ilkiv Department of Biochemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine 15:00 – 15:15 Application of novel polymeric nanocarrier for temozolomide delivery to glioblastoma cells Nataliya Finiuk

Department of Regulation of Cell Proliferation and Apoptosis, Institute of Cell Biology

National Academy of Sciences Ukraine, Lviv, Ukraine

15:15 – 15:30 Mapping of Residues of Fibrinogen Cleaved by Protease II of Bacillus thuringiensis var. israelensis IMV B-7465 Volodymyr Chernyshenko Palladin Institute of Biochemistry National Academy of Sciences Ukraine, Kyiv

15:30 – 15:45 Synthesis of potential antiviral agents among chromeno [4’,3’:4,5] thiopyrano[2,3-d][1,3]thiazole-5-carboxylates Nataliya Zelisko Department of Pharmaceutical, Organic and Bioorganic Chemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine

15:45 – 16:00 Panel Discussions Annual Review of the Research Activities in RECOOP

Moderators: Charles F. Simmons, Cedars – Sinai Medical Center, Los Angeles, CA, USA Iuliana Ceausu, “Carol Davila” University of Medicine and Pharmacy, Bucharest, Romania Peter Boor, RWTH University Aachen, Germany

16:00 – 17:00 Coffee Break Arts and Sciences Exhibit Conference Room 3 RECOOP Play Room Conference Room 2

Each artist – scientist has to introduce the artwork presented

17:00 – 18:00 Breakaway Sessions Session 1 Common Mechanism of Diseases – Low grade inflammation Congress Hall Chairs: Marija Heffer, Josip Juraj Strossmayer University of Osijek, Croatia Éva Szoko, Semmelweis University, Budapest, Hungary Peter Boor, RWTH University Aachen, Germany Oksana Zayachkivska, Danylo Halytsky Lviv National Medical University, Ukraine Oleksandr Korchynskyy, Institute of Cell Biology, NASU, Lviv, Ukraine Tamas Tabi, Semmelweis University, Budapest, Hungary Volodymyr Chernyshenko, Palladin Institute of Biochemistry NASU, Kyiv, Ukraine Participants: Anita Ćosic, Josip Juraj Strossmayer University of Osijek, Croatia Zrinka Mihaljevic, Josip Juraj Strossmayer University of Osijek, Croatia Marta Balog, Josip Juraj Strossmayer University of Osijek, Croatia Vedrana Ivic, Josip Juraj Strossmayer University of Osijek, Croatia Milorad Zjalic, Josip Juraj Strossmayer University of Osijek, Croatia Marta Skelin, University of Zagreb School of Medicine. Croatia Slavica Jurić Petričević, University of Split School of Medicine, Croatia Tereza Blahová, IKEM, Prague, Czech Republic Fruzsina Bagamery, Semmelweis University, Budapest, Hungary Judit Hajagos-Tóth, Faculty of Pharmacy, University of Szeged, Hungary Anita Sztojkov-Ivanov, Faculty of Pharmacy, University of Szeged, Hungary Kálmán Szucs, Faculty of Pharmacy, University of Szeged, Hungary Eva Nagy, University of Debrecen, Hungary Boglarka Donczo, University of Debrecen, Hungary Máté Szarka, University of Debrecen, Hungary Timea Kvarik, Medical School, University of Pécs, Hungary

Radana Gurecka, Comenius University, Bratislava, Slovakia Nazar Bula, Danylo Halytsky Lviv National Medical University, Ukraine Maryana Zvir, Danylo Halytsky Lviv National Medical University, Ukraine Jaroslav Pavlovskyi, Danylo Halytsky Lviv National Medical University, Ukraine Irena Pshyk-Titko, Danylo Halytsky Lviv National Medical University, Ukraine Khrystyna Malysheva, Institute of Cell Biology, NASU, Lviv, Ukraine Arsenii Borysov, Palladin Institute of Biochemistry NASU, Kyiv, Ukraine Iuliia Mazur, Palladin Institute of Biochemistry NASU, Kyiv, Ukraine Session 2 Nanobiomedicine – Drug conjugation Conference room 1 Chairs: Danylo Kaminskyy, Danylo Halytsky Lviv National Medical University, Ukraine Rostyslav Panchuk, Institute of Cell Biology, NASU, Lviv, Ukraine Ivan Alić, University of Zagreb School of Medicine, Croatia Lesya Kobylinska, Danylo Halytsky Lviv National Medical University, Ukraine Participants: Beata Zasonska, Institute of Macromolecular Chemistry AS CR, Prague, Czech Republic Maksym Moskvin, Institute of Macromolecular Chemistry AS CR, Prague, Czech Republic Maciej Chrzanowski, Wroclaw University of Technology, Poland Wroclaw Anna Lesiak, Wroclaw University of Technology, Poland Wroclaw Nataliya Finiuk, Institute of Cell Biology, NASU, Lviv, Ukraine Julia Senkiv, Institute of Cell Biology, NASU, Lviv, Ukraine Maryna Dudarenko, Palladin Institute of Biochemistry NASU, Kyiv, Ukraine Mariia Dekaliuk, Palladin Institute of Biochemistry NASU, Kyiv, Ukraine Ganna Pasichnyk, Palladin Institute of Biochemistry NASU, Kyiv, Ukraine Session 3 Contribution of perinatal viral infections to adverse outcomes of pregnancy in Central and Eastern Europe Conference room 6 Chair Shubhada Bopegamage, Slovak Medical University, Bratislava, Slovakia Participants: Pavel Bostik, Faculty of Military Health Sciences, University of Defense, Hradec Kralove, Czech Republic Brigita Benkoova, Slovak Medical University, Bratislava, Slovakia Andrijana Müller, Josip Juraj Strossmayer University of Osijek, Croatia Linn Defensor, Cedars – Sinai Medical Center, Los Angeles, CA, USA Veronika Puska, RECOOP HST Association, Budapest, Hungary

Session 4 RECOOP HST Association General Assembly Conference room 4 USA Cedars-Sinai Medical Center, Los Angeles, USA Edward Prunchunas, CSMC represented by Charles F. Simmons, Jr., MD and Sandor G. Vari, CSMC - RECOOP Croatia Josip Juraj Strossmayer University of Osijek, School of Medicine, Osijek, Croatia Ines Drenjancevic Josip Juraj Strossmayer University of Osijek, Department of Biology, Osijek, Croatia Elizabeta Has-Schon University of Zagreb School of Medicine, Croatia Srecko Gajovic University of Split School of Medicine, Department of Family Medicine, Split, Croatia Ivancica Pavlicevic Czech Republic IKEM - Institute for Clinical and Experimental Medicine, Prague, Czech Republic Jan Pitha Faculty of Military Health Sciences, University of Defense, Hradec Kralove, Czech Republic Pavel Bostik Hungary University of Debrecen, Hungary Absent University of Pecs, Hungary Tibor Ertl University of Szeged, Hungary Robert Gaspar Wroclaw University of Technology, Institute of Physics, Wroclaw, Poland Absent Carol Davila University of Medicine and Pharmacy, Bucharest, Romania Iuliana Ceausu Slovak Medical University, Bratislava, Slovak Republic Authorized representative Shubhada Bopegamage Ukraine Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kiev, Ukraine Tatiana Borisova Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv, Ukraine Rostyslav Stoika Danylo Halytsky Lviv National Medical University, Lviv, Ukraine Oleh Pinyazhko

18:00 – 18:40 Summary of the Breakaway Sessions 10 minutes for each session Congress Hall

Session 1 Common Mechanism of Diseases – Low grade inflammation Marija Heffer, Josip Juraj Strossmayer University of Osijek, Croatia Session 2 Nanobiomedicine – Drug conjugation Rostyslav Stoika, Institute of Cell Biology, NASU, Lviv, Ukraine Session 3 Contribution of perinatal viral infections to adverse outcomes of pregnancy in Central and Eastern Europe Shubhada Bopegamage, Slovak Medical University, Bratislava, Slovakia Session 4 RECOOP HST Association General Assembly Sandor G. Vari, CSMC - RECOOP 18:40 – 19:00 Summary of the Young Scientists Competition Sandor G. Vari, CSMC - RECOOP Veronika Puska, RECOOP HST Association, Budapest, Hungary 19:00 – 19:30 Closing Remarks

Sandor G. Vari, Director, International Research and Innovation Management Program, Cedars-Sinai Medical Center, Los Angeles, CA, USA & President of RECOOP HST Association

19:30 Dinner April 10, 2016 Departure

The Association for Regional Cooperation in the (RECOOP) Fields of Health, Science and Technology (HST)

Short Name: RECOOP HST Association

The Regional Cooperation for Health, Science and Technology (RECOOP HST) Consortium, led by Cedars-Sinai was formed in 2006 and transformed into Association in 2012 includes 17 universities and academic organizations from nine countries in Central and Eastern Europe (Croatia, Czech Republic, Hungary, Poland, Romania, Slovakia and Ukraine), Denmark and USA. The Association is a structured, functional and working research organization. According to its mission statement: "The RECOOP HST Association explores and enhances LOCAL scientific outputs of the partner organizations, creates critical mass of scientifically sound innovative research at REGIONAL level and exploits the research outcomes at GLOBAL level to improve the prevention and treatment of major public health problems."™

RECOOP builds multinational, multidisciplinary collaborations, and assists, coordinates the research activities of the eighteen research groups are the Cedars-RECOOP Research Centers (CRRC). Implementations of RECOOP’s strategic goals enable diverse talents geared towards integration of new knowledge derived from multispecialties to investigate Gender Differences (GD) in Common Mechanism of Diseases (CMD). In the CRRCs, researchers study genetic preconditions and the common mechanisms in molecular biology provide information on up-regulation or activation and down regulation or suppression of genetic codes by risk factors in nutrition, lifestyle (smoking, and alcohol, drug, mental and physical abuse), acute and chronic stress.

The scientific quality of the CSRRC’s research reflected in the RECOOP’s Annual Scientific Review Journals Biopolymers & Cell and Croatian Medical Journal.

Biopolym Cell. 2014;30(5 supplementary):1-4. http://www.biopolymers.org.ua/pdf/30/5suppl/001/biopolym.cell-2014-30-5suppl-001-en.pdf. Biopolym Cell. 2014;30(5 supplementary):5-44.

http://www.biopolymers.org.ua/pdf/30/5suppl/005/biopolym.cell-2014-30-5suppl-005-en.pdf. Biopolym Cell. 2013;29. http://www.biopolymers.org.ua/content/29/. Biopolym Cell. 2012;28. http://www.biopolymers.org.ua/content/28/. Biopolym Cell. 2011;27. http://www.biopolymers.org.ua/content/27/. Biopolym Cell. 2010;26. http://www.biopolymers.org.ua/content/26/. Croat Med J. 2014 Jun,55(3):181-286. http://www.cmj.hr/default.aspx?id=12345&issue=yes Croat Med J. 2015 Apr,56(2):75-176. http://www.cmj.hr/default.aspx?id=12482&issue=yes. Croat Med J. 2016 Apr, 57 (2) 87 -214 http://www.cmj.hr/default.aspx?id=12631&issue=yes The RECOOP HST Association organizes annually the Bridges in Life Sciences Conferences to review the scientific progress in the Association. During the Bridges in Life Sciences Annual Meeting the Scientific Advisory Board selects the top ten young scientists. The top ten young scientists selected during the Bridges in Life Sciences Annual Conferences have the opportunity to apply for International Visegrad Fund (IVF) Scholarship and receive the RECOOP Young Scientists Matching Fund. Also for the Bohdan Malaniak CSMC - RECOOP Young Scientists Research Award. The Visegrad Scholarship is the Visegrad Four European Macro-Region’s Fulbright Program. Therefore, it could be important to link the Visegrad Scholarship and the Fulbright Foreign Student Program. CSMC – RECOOP Research Centers (CRRC) are the Center of Excellences of the RECOOP HST Association. They host young scientists, Ph.D. students with CSMC – RECOOP (IVF – CSMC - RECOOP) Scholarship. The RECOOP HST Association Scientific Advisory Board selects the young scientists who could apply for IVF – CSMC - RECOOP Scholarship. The selected young scientists (preferably Ph.D. students) will spend maximum four semesters receiving: €2,300 / semester and the corresponding host universities/institutes will receive €1,500/semester/scholar. The host CRRC will get €1,000 for laboratory expense and consumables from CSMC – RECOOP HST Association. The Bohdan Malaniak CSMC - RECOOP Young Scientists Research Award for preclinical and clinical studies. The host and appointing organizations can be basic or translational research organizations nevertheless they should have an assisting clinical organization with clinical practice. The research project and the performed lab tests and measurements must have clinical relevance. The maximum requested amount is 1,500 USD but it will only be paid if the applicant provides and proves matching found for the same amount. The submission date July 27, 2016, the application should be sent to Sandor G. Vari, MD. Please follow strictly the word counts in the template! RECOOP would like to provide more opportunities for young scientists, therefore agreed with Korányi Frigyes Science Dormitory, Semmelweis University Budapest to select the top abstracts were submitted to the call of Korányi Frigyes Scientific XXI Forum of University Semmelweis, Budapest, Hungary. The Forum was on March 10 -11, 2016, and aimed to provide opportunities for young researchers, clinicians, pharmacists and students involved in Students’ Scientific Association, while promoting a scientific contact between various disciplines within our university. The submitted 22 English abstracts were reviewed by RECOOP Review Board for the Bridges in Life Sciences 11th Annual Conference, Prague, Czech Republic. This peer –review is

considered by RECOOP as a pre-selection, and will be finalized after the outcome of the Koranyi Science Forum 2015. All abstracts were reviewed by minimum 2 RECOOP experts. Those who had no ECA - Ethical Committee or IACUC – Institutional Animal Care and Use Committee approvals and could not or did not provide the approval were excluded from the final selection. Only those received RECOOP invitation for the Bridges Conference in Prague who were already selected by RECOOP Review Board and at the same time ranked first or second in their session during the 2016 Koranyi Science Forum.

Andras Makkos Medical Student VI Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary Tim Frederik Svanya Medical Student V. Department of Physiology, Faculty of Medicine, Semmelweis Universtiy Orsolya Szabo Medical Student V. Institute of Microbiology, Faculty of Medicine, Semmelweis University

The Head of the Korányi Frigyes Scientific XXI Forum of Organizing Committee Klára Alíz Stark, 3rd year Medical Student, Faculty of Medicine, Semmelweis University, Budapest, Hungary received an invitation as an appreciation of her hard work she is displaying her art work in the Arts and Sciences Exhibit. The RECOOP HST Association with Korányi Frigyes College for Advanced Studies, Semmelweis University, Budapest, Hungary (http://harsfa.semmelweis.hu/recoop-fkcas-lsiiaw.php ) announced the first annual Art and Sciences Competition. The main goal of the RECOOP Annual Art and Sciences Competition is to initiate a creative working relation between RECOOP young scientists and artists to create artworks from life science and medical images and communicate the beauty of the living organism. Dr. Sandor G. Vari, MD Director, International Research and Innovation Management Program Cedars-Sinai Medical Center & President of the RECOOP HST Association 6420 Wilshire Blvd., Ste. 300, Los Angeles, CA 90048-5502, USA Office 1: 1 323-866-8122; 1 323-866 -6824 Office 2: 1 818 904 1954 Mob: 1 818 398-2642 E-mail: vari@cshs.org

The Croatian Medical Journal (April 2016, www.cmj.hr) published now the third RECOOP thematic issue, this time dedicated to the RECOOP Bridges 11th Annual Scientific Conference in Prague, Czech Republic.

The mission of Croatian Medical Journal (CMJ) was from its very beginning to support

the biomedical research in emerging countries. CMJ first aimed to help Croatian research community to publish their high quality papers in a friendlier way, but very soon, the scope was extended to the neighboring countries, Central East Europe, Asia and Africa. Currently, CMJ is orientated to all researchers having low visibility either due to their geographical location or due to their professional status.

The major tool that CMJ is offering to this group of emerging researchers is a specific Author-Helpful Policy. The submitted articles were judged according to their potential and the dedicated team of professionals is there to assist the authors. The team consisted of the members of CMJ editorial group and CMJ associates were engaged to help authors to present their research according to the high standards of international publishing. Such policy had many benefits. The first were for the Journal itself, as it helped to establish a core of knowledge under CMJ umbrella which although offered internationally was indeed a national asset. Croatian experts in statistics, manuscript editing, scientific language, and manuscript production were established. The criteria of manuscript quality subsequently raised and the CMJ Impact Factor increased to its maximum of 1.8, which made it to rank among first 10 general medical journals.

The effects on global community were as well visible. Many young researchers were able to publish their first contributions in CMJ, many other national journals followed the suite and the strength of the previously unrecognized research became obvious and accessible to the global scientific community.

CMJ has published the second thematic issue made by scientists from RECOOP institutions. We can say that there is already a small tradition to offer and share CMJ virtues with the RECOOP scientists and to collaborate successfully.

We at CMJ strongly believe that RECOOP and CMJ share their basic mission. By increasing the across border collaboration in the Central and Eastern Europe the undiscovered opportunities start to shine and bring benefits to the national, regional and global community. Indeed, the scientific community should continuously discover multinational and multidisciplinary collaborations, and should be nourished, developed and properly presented.

The whole venture to create this thematic issue was an excellent international team effort, were many small pieces were arranged together to identify, develop and present the excellence growing under RECOOP umbrella. Therefore, we wish that the published papers in the thematic issues are a clear sign of growing common RECOOP activities, and a long-term establishment of regional excellence, excellence that CMJ wants to be part of.

Prof. Srecko Gajovic Editor-in-Chief Croatian Medical Journal CMJ is an international peer-reviewed Diamond Open Access journal published six times per year. The CMJ uses the Diamond Open Access model. This means that there are NO author processing fees and NO fees to access the published papers. The free access is available on our web page, www.cmj.hr, also on PubMed Central. Impact factor of CMJ for 2013 is 1.373. The RECOOP thematic issue is as well fully accessible at journal web page www.cmj.hr

Abstracts

Abstracts of Common Mechanism of Diseases Diet induced obesity and impaired glucose metabolism in elderly rats treated with Liraglutide (Victoza) and Metformin. 1Gaspar R, 1Ducza E, 1Seres AB, 1Szűcs KF, 1Hajagos-Tóth J, 1Sztojkov-Ivanov A, 1Tiszai Z 1Minorics R, 1Schelz Zs, 1Bóta J, 1Csányi A, 2Vari SG 1Department of Pharmacodynamics and Biopharmacy, University of Szeged, Hungary 2International Research and Innovation in Medicine Program, Cedars - Sinai Medical Center, Los Angeles, CA, USA and Regional Cooperation for Health, Science and Technology (RECOOP HST) Association, Debrecen, Hungary Corresponding author: Robert Gaspar gaspar@pharm.u-szeged.hu Key words: high calorie diet, obesity, glucose tolerance, elderly rat, liraglutide, metformin Introduction: Obesity is a major public health concern. Our aim was to develop an animal model for obesity in aging rats by diet. We intended to measure -among others- the obesity-induced changes in body weight, glucose metabolism, cognitive and renal functions and uterine contraction. Methods: Male and female rats (44-weeks-old) were put in 4 groups containing 8 male and 8 female rats each: 1. Standard diet (SD); 2. High fat-high sucrose diet (HFHSD); 3. HFHSD+ metformin (50 mg/kg/day); 4. HFHSD+liraglutide (0.3 mg/kg/day) from 51 weeks for 14 weeks of ages. Obesity was induced by HFHSD by from 45 weeks of age for 20 weeks. Animals were sacrificed at their 64 weeks of age. Obesity was evaluated by body weight and waist circumference; glucose metabolism was measured by glucose tolerance and insulin sensitivity tests, cognitive function was tested by water maze, renal function was measured by urine tests, uterine contractility was measured in organ bath in vitro. Results: HFHSD induced obesity. The gain in weight was significantly reduced by liraglutide in both sexes, while metformin slightly reduced the bodyweight only in males. Obesity induced an impaired glucose metabolism in both sexes that was reduced by metformin and liraglutide. A slight reduction in cognitive function was detected in male rats. The majority of the urine tests shown inconsistency, slight increases in microalbuminuria (HFHSD males) and glucose (HFHSD females) were detected. Discussion and Conclusion: Long-term HFHSD induced obesity and impaired glucose metabolism in elderly rats and affected cognitive and renal functions. Liraglutide was more effective to restore the consequences of obesity. Our animal model seems to be a highly identical experimental system to induce the process and consequences of human obesity. Sources of Funding: This work has been supported by RECOOP HST grant. Acknowledgement: The study was supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and the participating Cedars – Sinai Medical Center - RECOOP Research Centers (CRRC). Ethical Committee Approval: All experiments were carried out with the approval of the Hungarian Ethical Committee for Animal Research (registration number: IV/3796/2015).

Glucose and insulin tolerance test in obese, elderly rats with impaired glucose metabolism treated with Liraglutide (Victoza) and Metformin

1Szűcs KF, 1Ducza E, 1Seres AB, 1Tiszai Z, 2Vari SG, 1Gaspar R, 1Department of Pharmacodynamics and Biopharmacy, University of Szeged, Hungary 2International Research and Innovation in Medicine Program, Cedars - Sinai Medical Center, Los Angeles, CA, USA and Regional Cooperation for Health, Science and Technology (RECOOP HST) Association, Debrecen, Hungary Corresponding author: Robert Gaspar gaspar@pharm.u-szeged.hu Key words: obesity, glucose tolerance, elderly rat, liraglutide, metformin Introduction: The increasing prevalence of obesity is a major public health concern. Obesity is responsible for the development of diabetes mellitus, arteriosclerosis, hypertension and several other diseases. Our aim was to develop an animal model for obesity in aging rats by diet. We intended to measure the obesity-induced changes in glucose metabolism and insulin sensitivity. Methods: Male and female rats (44-weeks-old) were put in 4 groups containing 8 male and 8 female rats each: 1. Standard diet (SD); 2. High fat-high sucrose diet (HFHSD); 3. HFHSD+ metformin (50 mg/kg/day); 4. HFHSD+liraglutide (0.3 mg/kg/day) from 51 weeks for 14 weeks of ages. Obesity was induced by HFHSD by from 45 weeks of age for 20 weeks. Glucose tolerance was measured at 44, 50, 57 and 63 weeks of age, while insulin sensitivity test was carried out at 64 weeks of age with OneTouch® UltraMini® Glucose Meter (Milpitas, CA, USA). Results: HFHSD induced impaired glucose tolerance after 5 weeks of diet that were increased gradually every month. In females, the aging itself induced worsening of glucose tolerance. Liraglutide restored the worsening glucose tolerance after 7 weeks of treatments while metformin was able to moderate the process, only. However, the glucose tolerance increasing effect of the liraglutide was disappeared on 13th week of treatment while metformin maintained its moderating effect especially in females. The insulin sensitivity test before the sacrifice revealed that liraglutide slightly increased the sensitivity in males, while metformin was effective in females. Discussion and Conclusion: Long-term HFHSD induced impaired glucose tolerance in elderly rats. Liraglutide is able to restore the glucose tolerance while Metformin is less effective in 7-week therapy. However, 13-week treatment caused benefit cease of liraglutide therapy. Sources of Funding This work has been supported by RECOOP HST grant. Acknowledgement: The study was supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and the participating Cedars – Sinai Medical Center - RECOOP Research Centers (CRRC). Ethical Committee Approval: All experiments were carried out with the approval of the Hungarian Ethical Committee for Animal Research (registration number: IV/3796/2015).

Alteration of inflammatory markers and SSAO activity in diet induced diabetic, obese elderly rats treated with Liraglutide (Victoza) and Metformin Tábi T1, Szökő É1, Gáspár R2, Heffer M3, Vári SG4… 1Department of Pharmacodynamics, Semmelweis University, Hungary 2Department of Pharmacodynamics and Biopharmacy, University of Szeged, Hungary 3Department of Medical Biology, University of Osijek, Osijek, Croatia 4International Research and Innovation in Medicine Program, Cedars - Sinai Medical Center, Los Angeles, CA, USA Corresponding author: Tamas Tabi abi.tamas@pharma.semmelweis-univ.hu Keywords: obesity, diabetes, low grade inflammation, inflammatory cytokines Introduction: The unhealthy lifestyle is one of the major risk factors of metabolic disorders such as obesity, insulin resistance and type 2 diabetes. Accumulation of excess adipose tissue especially in the visceral fat depots accompanies to a low grade inflammatory state and increased production of proinflammatory cytokines and adipokines. These mediators in turn can interfere with insulin signaling triggering insulin resistance. Aims: In the present study we aimed at characterizing the metabolic parameters and adipose tissue inflammation in a high fat, high sugar diet induced model of obesity in elderly rats. Effect of gender and antidiabetic therapies was also evaluated. Results: Diet induced obesity was accompanied by fasting hyperinsulinemia reaching significance only in case of male animals due to the high variance in the obese group. These animals can be divided to two groups; ones with normal or slightly elevated insulin levels and ones with considerable hyperinsulinemia suggesting the importance of genetic predisposition factors. Plasma leptin levels showed a similar profile with significantly higher leptin level in females. Adiponectin levels were unaltered. Inflammatory cytokines (TNFα, IL-1β, IL-6) concentrations were altered mainly in the visceral fat. Their levels were generally increased in obese male animals while females showed elevated concentration inherently with no further increase in the obese animals. SSAO activity in the visceral fat was unaltered and reduced in obese males and females, respectively suggesting the enzyme plays no significant role in adipose tissue inflammation. Antidiabetic therapies had only minor effects on the examined parameters. Liraglutide generally showed more favorable effects compared to metformin. Conclusion: High fat, high sugar diet induced metabolic changes characteristic to type 2 diabetes but a portion of animals showed resistance to the effect of the diet. Inflammation in the adipose tissue of elderly females was more pronounced while unhealthy diet has higher effect in males that is in accordance with the higher risk of postmenopausal females. Ethical approval: All experiments were carried out with the approval of the Hungarian Ethical Committee for Animal Research (registration number: IV/3796/2015). Acknowledgement: This study was supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association).

Leptin and insulin receptor expression in the brain of diet induced diabetic, obese elderly rats/ treated with Liraglutide (Victoza) and Metformin Senka Blažetić1, Irena Labak1, Marija Heffer2, Rober Gaspar3, Sandor Vari4 1 Department of Biology, University J. J. Strossmayer of Osijek, Osijek, Croatia; 2 Department of Medical Biology, School of Medicine, University J. J. Strossmayer of Osijek, Osijek, Croatia; 3 Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, Eötvös u. 6. H-6720 Szeged, Hungary, 4 International Research and Innovation in Medicine Program, Cedars-Sinai Medical Center, Los Angeles, CA, United States of America or just USA and Regional Cooperation for Health, Science and Technology (RECOOP HST) Association, Debrecen, Hungary Corresponding author e-mail address: mheffer@mefos.hr Key words: obesity, diabetes, insuline resistance, leptin resistance, Liraglutide, Metformin Introduction: Advancing age and obesity are the major risk factors for development of diabetes type 2. Occurrence of insulin resistance in brain is named ‘diabetes type 3’ and probably contributes to cognitive defect in Alzheimer. Anti-diabetic drugs, like Metformin and Liraglutide, may have different capacity to treat central insulin resistance. Method: Twenty week old male and female SD rats were divided in: control group and 3 groups fed high fat-high sugar diet (HSHFD). From fifty first week one HSHFD group was treated with Metformin (50 mg/kg/day s.c.) and another one with Liraglutide (0.3 mg/kg/day s.c). All groups were sacrificed at 65 weeks of age. Brain was fixed and immunostained for insulin (IR) and leptin (ObR) receptor. Results: Compared with control group, male rat groups on HSHFD had significantly higher expression of IR in hippocampus, while female group had decreased level of IR (P = 0.04). HSHFD generally reduced expression of ObR in hypothalamus. Both drugs, Metformin and Liraglutide increased hypothalamic ObR expression during HSHFD. Liraglutide had stronger effect in female group. Discussion – conclusion: It is well known that insulin resistance is related to obesity. Results indicate that there are gender differences in insulin signaling pathway. IR in brain supports cognitive functions, including learning and memory and it decreases in aging. Based on idea that reduced IR results with synapse loss it is possible that female group on HSHFD, more likely than male group, develop cognitive defect. Low levels of ObR in hypothalamus in male and female groups accompanies obesity. According to our results the HSHFD affects IR and ObR distribution in the brain. Also, Metformin and Liraglutide had sex specific capacity to treat central insulin resistance. Sources of Funding: This work has been supported in part by VIF-MEFOS-1 (Faculty of Medicine Osijek, Croatia) and by RECOOP HST grant. Ethical Approval: All experimental procedures are conformed to the European Guidelines for the Care and Use of Laboratory Animals (directive 86/609). They were approved by the by Hungarian Ethical Committee for Animal Research (IV/3796/2015). Acknowledgement: The study was supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and the participating Cedars – Sinai Medical Center - RECOOP Research Centers (CRRC).

Markers of Blood Oxidative Stress and Antioxidative Enzymes Activity in Obese Diabetic Elderly Rats Treated with Metformin or Liraglutide Drenjancevic, Ines1; Cosic, Anita1; Vuković, Rosemary2; Kljajić, Julijana2; Heffer, Marija 3; Gaspar, Robert 4; Vari, G Sandor 5 1- University Josip Juraj Strossmayer Osijek, Faculty of Medicine, Department of Physiology and Immunology, Cara Hadrijana 10e, 31000 Osijek, Croatia, Osijek, Croatia, 2- University Josip Juraj Strossmayer Osijek, Department of Biology, Osijek, Croatia, 3- University Josip Juraj Strossmayer of Osijek, Faculty of Medicine Osijek, Department of Medical Biology and Genetics, Cara Hadrijana 10e, 31000 Osijek, Croatia, 4- Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, Eötvös u. 6. H-6720 Szeged, Hungary, 5-, International Research and Innovation in Medicine Program, Cedars-Sinai Medical Center, Los Angeles, CA, United States of America or just USA and Regional Cooperation for Health, Science and Technology (RECOOP HST) Association, Debrecen, Hungary The corresponding author: Ines Drenjancevic; ines.drenjancevic@mefos.hr Key words: antioxidative enzymes activity, TBARS, FRAP, diabetes, obesity, Introduction: Diabetes mellitus and obesity present with increased oxidative stress. Metformin increases insulin sensitivity and influences glucose metabolism in liver and in peripheral tissues. Liraglutide is a glucagon-like peptide-1 receptor agonist that stimulates insulin secretion.This study aimed to determine the markers of oxidative stress in diabetic obese elderly Sprague-Dawley (SD) rats treated with metformin or liraglutide. Methods: Male and female SD rats were divided in: 1) control group; 2)group fed high fat-high sucrose diet from 20 to 65 weeks of age (HSHFD); 3) HSHFD+Metformin (50 mg/kg/day s.c.); and 4) HSHFD+Liraglutide (0.3 mg/kg/day s.c). Drugs were given concomitantly to HSHFD from 51-65 weeks of age when rats were sacrificed. Plasma lipid peroxidation products were assessed by TBARS and antioxidative capacity by FRAP. SOD, catalase and gluthatione peroxidase activity was assessed by spectrophotometry. Results: Females have no difference in TBARS among groups. FRAP was significantly higher in HFHSD+Liraglutid compared to HFHSD+Metformin and HSHFD. Catalase activity was higher in control compared to other female groups. TBARS and FRAP were similar among male groups. Control group had higher catalase activity compared to other male groups; higher GPx activity compared to HSHFD+metformin and HSHFD+liraglutide and higher SOD activity compared to HSHFD+metformin. Comparing Female vs. Male showed female control rats had significantly higher TBARS compared to male control.Catalase activity was significantly decreased in all diet groups of both sexes compared to respective control groups and decreased in female groups overall, compared to male. Discussion: Metformin significantly decreased all antioxidative enzymes activity compared to control in male. Treatment with liraglutide increased antioxidant capacity compared to HFHSD+metformin and HFHSD groups in female to control levels, but oxidative stress was not significantly changed. Conclusion: There are sex-related differences in the level of oxidative stress and antioxidative enzymes activity. Antidiabetic drugs may modify antioxidative capacity more in female than male. Metformin and liraglutide decrease activity of antioxidative enzymes in male. Sources of Funding This work has been supported in part by VIF-MEFOS-15 (Faculty of Medicine Osijek, Croatia) and by RECOOP HST grant. Ethical Approval All experimental procedures are conformed to the European Guidelines for the Care and Use of Laboratory Animals (directive 86/609). They were approved by the by Hungarian Ethical Committee for Animal Research (IV/3796/2015). Acknowledgement: The study was supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and the participating Cedars – Sinai Medical Center - RECOOP Research Centers (CRRC).

Fatty acid changes in plasma, visceral and subcutaneous adipose tissue in diabetic, obese elderly rats treated with Liraglutide (Victoza) and Metformin. In memoriam of Janos Harangi, University of Debrecen, Hungary Nagy E1, Krivosikova Z2, Gajdos M2, Kebis A3, Vari SG4, Tábi T5

. 1Doctorate School of Animal Husbandry, Faculty of Agriculture and Nutritional Sciences, University of Debrecen, Hungary 2Department of Clinical and Experimental Pharmacotherapy, Slovak Medical University, Medical Faculty, Bratislava, Slovakia 3Department of Toxicology, Faculty of Public Health, Slovak Medical University, Bratislava, Slovakia 4International Research and Innovation in Medicine Program, Cedars - Sinai Medical Center, Los Angeles, CA, USA 5Department of Pharmacodynamics, Faculty of Pharmacy, Semmelweis University, Hungary Corresponding author: Dr. Tamás Tábi, PhD tabi.tamas@pharma.semmelweis-univ.hu Keywords: fatty acid composition, high fat diet, physical exercise Introduction: Adipose tissue contains triglycerides of various fatty acids. Among them the most abundant saturated and monounsaturated ones serve mainly as energy storage while the polyunsaturated ones are the precursors of a range of mediators such as prostanoids and leukotrienes are potent inflammatory mediators. Significant evidence has been accumulated to support the cardiovascular protective role of n-3 fatty acids compared to n-6 ones. Aims: In this study we aim to develop a sample preparation method to analyzing fatty acid composition in visceral and subcutaneous adipose tissue of rats in response to diet and physical activity. Methods: High-fat diet was used to induce obesity in rats. A subgroup of rats was forced to running to mimic physical exercise while another subgroup was also fed by antioxidant selenized cookie. In the sample preparation step we tried more acid catalyzed and base catalyzed transesterification method before we developed the appropriate one. Fatty acid composition of white adipose tissue was measured by gas chromatography. Results: The first results showed that high-fat diet significantly increased the saturated and monounsaturated fatty acid content, while the polyunsaturated fatty acid content decreased significantly in both tissues contrary its relatively high intake. These fatty acids are included in very low concentration in the tissue and in some cases their amounts were under the detection limit. The method development shows that we need a higher concentration step to be able to determine and compare these essential fatty acids. Conclusion: Our data shows that we need acid catalyzed derivatization followed by a strong concentration step in the sample preparation. In this way we will be able to follow up the diet- induced changes in the body. Ethical approval: The proposal was approved by the Ethical Committee for Animal Experiment of the Slovak Medical University and by the State Veterinary and Food Authority of the Slovak Republic (number of approval: Ro-1651/11-221b, 7.10.2011). Acknowledgement: This study was supported by Cedars Sinai Medical Center's International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and "Center of excellence of environmental health" project, ITMS No.26240120033, based on the supporting operational Research and development program financed from the European Regional Development Fund."

Renal pathological alterations in diet induced diabetic, obese elderly rats treated with Liraglutide and Metformin B.M. Klinkhammer1, P. Boor1 R Gaspar2, S. G. Vari3 1 Institute for Pathology, University hospital RWTH Aachen, Germany 2Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, Eötvös u. 6. H-6720 Szeged, Hungary, 3International Research and Innovation in Medicine Program, Cedars-Sinai Medical Center, Los Angeles, CA, USA and Regional Cooperation for Health, Science and Technology (RECOOP HST) Association, Debrecen, Hungary Corresponding Author: Peter Boor pboor@ukaachen.de Key words: metabolic syndrome; kidneys, histology, Introduction The metabolic syndrome is a major world-wide public-health problem associated with lifestyle. Metabolic syndrome promotes development of co-morbidities like diabetes, cardiovascular diseases and chronic kidney disease. In this experimental study we analyze the effects of a glucagon-like peptide-1 (GLP-1) mimetic Liraglutide and the classical anti-diabetic drug Metformin on obesity-induced kidney disease. Methods Nine month ole female and male rats were kept on normal diet (control group) or fed with a high fat high sugar diet (HFHSD) for 19 weeks. Starting at week 6 of HFHSD diet, the chow was supplemented either with Liraglutide (0.3mg/kg) or Metformin (50mg/kg). Control animals received pure HFHSD or continued to receive normal rat chow. Body weight was measured weekly and blood glucose tolerance was measured monthly. At the end of experiment, plasma and urine was collected and waist circumference and kidney weights were recorded. Kidneys were harvested for histological, immunohistochemical and ultrastructural analyses. Results No difference in kidney weights was observed. Histologically, kidneys showed focal pathological glomerular changes, e.g. thickening of basement membrane with subendothelial exudates, mesangial expansion and sclerotic lesions. Additionally, activation of parietal epithelial cells and tubular cells was observed in CD44 staining, a marker of injury-associated epithelial cell activation in kidneys. PAS staining showed focal tubular injury and atrophy, including loss of brush-border, tubular dilation and cellular simplification, and thickening of tubular basement membranes. Associated to these changes interstitial mononuclear inflammation was observed. Immunohistochemistry showed increased number of interstitial aSMA positive myofibroblasts, a marker of activated fibroblasts, and increased deposition of collagen type I confirming the presence of tubulointerstital fibrosis. No obvious changes between the different groups on HFHSD were observed, requiring further detailed histomorphological analyses and quantification. Discussion Kidneys from elderly rats with HSHFD showed obvious morphological renal changes. Future plan To evaluate differences between the treatment groups, we will perform histomorphological quantification of the observed kidney damage, i.e. tubulointerstitial injury and glomerular sclerosis (in PAS), activation of parietal epithelial cells (CD44), podocyte damage (Desmin), interstitial and glomerular inflammation (CD68, CD44), interstitial fibrosis and glomerular sclerosis (Collagen I, IV), interstitial myofibroblasts (aSMA). Ultrastructural changes, in particular of the glomeruli, will be analyzed with transmission electron microscopy. Kidney function, e.g. serum creatinine and BUN, and proteinuria/albuminuria and tubular injury, e.g. using urinary KIM-1 or NGAL ELISA, should be assessed in blood and urine samples. Ethical Approval: All experimental procedures are conformed to the European Guidelines for the Care and Use of Laboratory Animals (directive 86/609). They were approved by the by Hungarian Ethical Committee for Animal Research (IV/3796/2015). Acknowledgement: The study was supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and the participating Cedars – Sinai Medical Center - RECOOP Research Centers (CRRC).

Altered uterine contractility and response to adipokines after Liraglutide (Victoza) and Metformin treatment in obese, elderly rats 1Gaspar R, 1Hajagos-Tóth J, 1Ducza E, 1Tiszai Z, 1Bóta J, 1Csányi A 2Vari SG 1Department of Pharmacodynamics and Biopharmacy, University of Szeged, Hungary 2International Research and Innovation in Medicine Program, Cedars - Sinai Medical Center, Los Angeles, CA, USA and Regional Cooperation for Health, Science and Technology (RECOOP HST) Association, Debrecen, Hungary Corresponding author: Robert Gaspar gaspar@pharm.u-szeged.hu Key words: high calorie diet, obesity, elderly rat, liraglutide, metformin, uterine contractility Introduction: Obesity in pregnancy is associated with failure in uterine contractions. Other studies reported that maternal overweight and obesity during pregnancy were associated with increased risks of preterm delivery. Some adipokines that have increased levels in obesity reduce the contractility of human or rat myometria. Our aim was to investigate how obesity alters the non-pregnant uterine contractility and its response to leptin and adiponectin. Methods: Female rats (44-weeks-old) were put in 4 groups containing 8 rats each: 1. Standard diet (SD); 2. High fat-high sucrose diet (HFHSD); 3. HFHSD+ metformin (50 mg/kg/day); 4. HFHSD+liraglutide (0.3 mg/kg/day) from 51 weeks for 14 weeks of ages. Obesity was induced by HFHSD by from 45 weeks of age for 20 weeks. Animals were sacrificed at their 64 weeks of age. Spontaneous and KC-induced uterine contractility was measured in organ bath in the presence of leptin (10-7M) or adiponectin (10-8M). The mRNA expressions of uterine leptin and adiponectin receptors were detected by RT-PCR. Results: Neither obesity nor metformin or liraglutide treatment changed the spontaneous contractions. The KCl-induced contractions were reduced by both metformin and liraglutide treatment. Leptin and adiponectin increased the non-obese, while reduced the obese spontaneous contractions. The relaxation was enhanced by metformin treatment. Leptin slightly reduced the KCl-induced contraction bot in non-obese and obese uteri. Liraglutide reversed the action of leptin to contraction. Adiponectin had no effect on KCl-induced contraction, while metformin or liraglutide treatment enhanced its action to relaxation. Discussion and Conclusion: Obesity alters the non-pregnant uterine contractility, metformin and liraglutide treatment can modify the action of leptin and adiponectin on contraction of obese rat uterus. Sources of Funding This work has been supported by RECOOP HST grant. Acknowledgement: The study was supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and the participating Cedars – Sinai Medical Center - RECOOP Research Centers (CRRC). Ethical Committee Approval: All experiments were carried out with the approval of the Hungarian Ethical Committee for Animal Research (registration number: IV/3796/2015).

Bone mineral density and content in elderly diabetic rats. The impact of Metformin and Liraglutide treatment. Krivošíková Z1*, Štefíková K1, Heffer M2, Gáspár R3,Vari SG4. 1Department of Clinical and Experimental Pharmacotherapy, Medical Faculty, Slovak Medical University, Bratislava, Slovakia 2Department of Medical Biology, School of Medicine, University J. J. Strossmayer of Osijek, Osijek, Croatia 3Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, Eötvös u. 6. H-6720 Szeged, Hungary 4International Research and Innovation in Medicine Program, Cedars-Sinai Medical Center, Los Angeles, CA, United States of America or just USA and Regional Cooperation for Health, Science and Technology (RECOOP HST) Association, Debrecen, Hungary * zora.krivosikova@szu.sk Key words: rat, diabetes, bone, Metformin, Liraglutide Introduction: The effects of type 2 diabetes on bone are not clear yet. Whereas bone mineral density (BMD) has been reported to be either unchanged or modestly increased, incidence of low-stress fractures is almost doubled together with delayed fracture healing. In vitro data indicates that metformin can directly stimulate the proliferation, differentiation and mineralization of osteoblasts and liraglutide has anabolic effects in young diabetic rats. Our objective was therefore to determine the potential positive effect of Metformin and Liraglutide treatmenton femoral and tibialBMD and bone mineral content (BMC) in rats with diabetes induced by high fat-high sugar diet (HFHSD). Methods: Twenty week old male and female SD rats were divided in: control group and 3 groups fed HSHFD. From fifty first week one HSHFD group was treated with Metformin (Met; 50 mg/kg/day s.c.) and another one with Liraglutide (LG; 0.3 mg/kg/day s.c). All groups were sacrificed at 65 weeks of age. Standard biochemistry was measured by biochemical analyzer (Vitros 250, J&J, Rochester, USA). Dual X-ray absorptiometry wascarried out by using a Lunar Prodigy Advance with Encore 2011 software version 13.60 (GE Medical Systems, Madison, WI, USA). Results: The bone marrow adiposity was similar in both genders. The excess of adipocytes was found in HFHSD group and LG groups if compared to SD group, and much less in Met group if compared to HFHSD group. No significant differences were found in length, BMD, BMC and area of right femur and right tibia in all experimental groups in both female and male gender. Strong correlation was confirmed between bone lengths and BMC or area in female rats and between bone lengths and BMD, BMC and area in male rats. No correlation was observed between densitometrical data and biochemical parameters in both genders. Discussion and conclusion:All the available literary data on the detrimental effects of obesity and diabetes on bone, and the potential anabolic effects of antidiabetic agents have been described in growing young and adult rats. In our experiment, the animals were subjected to HFHSD at an advanced age with sufficient peak bone mass. We concluded that obesity and diabetes affect probably more bone formation than bone resorption. Elderly rats have already been subdued bone formation and therefore the impact of diabetes and antidiabetics treatment on bone mineral was not shown. Sources of research support: This work has been supported by RECOOP HST Association research grant and by the frame work of realization of the project "Center of excellence of environmental health", ITMS No.26240120033, based on the supporting Operational Research and Development Program financed from the European Regional Development Fund. Acknowledgement: The study was supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and the participating Cedars – Sinai Medical Center - RECOOP Research Centers (CRRC). Ethical Committee Approval: All experiments were carried out with the approval of the Hungarian Ethical Committee for Animal Research (registration number: IV/3796/2015).

Abstracts of Cardiovascular Research in Progress ORAL PRESENTATIONS

Cardiovascular risk factors and their impact on vascular parameters in women are modified by reproductive status and smoking, longitudinal study. J. Pitha, I. Kralova Lesna, P. Wohlfahrt, O. Auzky, A. Sekerkova, M. Lejsková, P. Stavek, R. Dembovská, J. Mrázková, R. Houdková; Center for Experimental Medicine, Institute for Clinical and Experimental Medicine, Prague Corresponding author: Jan Piťha japi@ikem.cz Key words: reproductive status – cardiovascular risk factors – remnant lipoproteins - vascular biomarkers Introduction: In our previous studies we detected increased sensitivity for development and progression of atherosclerosis in women in menopausal transition especially when smoking. Additionally, we observed strong association between preclinical atherosclerosis and remnant lipoprotein particle cholesterol (RLP-C). The main aim of this study was to analyze impact of changes of cardiovascular risk factors under study on several established vascular parameters. Methods: We analyzed changes of main cardiovascular risk factors including RLP-C (total minus LDL minus HDL cholesterol in fasting status) in 10year longitudinal study and the effect of these changes on vascular parameters including aortic stiffness, arterial stiffness, endothelial dysfunction and circulating microparticles measured at the end of the study. Results: In 361 women aged 59.9 ± 2.6 years we detected unfavorable changes of almost all risk factors during the study period. In these changes were not substantial different between women who changed their status to menopause and permanently menopausal women. RLP-C were negatively associated with endothelial function in the whole population (r=-0.133, p

Shared decision making in life style and nutrition for intervention in woman with risk factors in cardiovascular health Jurić Petričević S1, Pavličević I2, Malički M3, Mrduljaš-Đujić N2, Ćurčić L2, Došen Janković S4, Škrabić S5, Tolić-Biočina A5, Marušić M3, Marušić A3

1Department of Emergency Medicine in Split, Department of Family Medicine, University of Split School of Medicine, Šoltanska 2, Split, Croatia 2Department of Family Medicine, University of Split School of Medicine, Šoltanska 2, Split, Croatia 3Department of Research in Biomedicine and Health, University of Split School of Medicine, Šoltanska 2, Split, Croatia 4Family practice office, Put Skalica 2, Split, Croatia 5Family practice office, Getaldićeva 27, Split, Croatia The corresponding author: Slavica Jurić Petričević, slavica.juric01@gmail.com Key words: woman, cardiovascular health, life style, nutrition Introduction: In 2015 we conducted an interventional study to decrease CVD risk in women between 40 and 65 years of age. After 6 months, 33% of women decrease their risk. In order to determine their satisfaction with the intervention, and possible way of improving it, we conducted a survey on all women participating in the trial. Methods: Prospective, interventional study included 95 women (7 patients dropped out of study) of different reproductive status and one or more of cardiovascular risk factors. We divided patients in 3 groups in accordance with the results of cardiovascular disease risk (CVD) change: those who decreased CVD risk, those with no improvement and those who increased CVD risk, and analyzed their answers. Results: Of the 95 women 81 (91%) reported attempting to change their life habits, and 28 (61%) tried to stop smoking (out of 46 who smoked). However, only 3 actually stopped smoking. Allmost all (91) found the intervention to be usefull for them. The women who did not change or who increased the CVD risk after the intervention had higher hope than those who were successful. It is possible these less successful women would have benefited from the prolonged and more intensive intervention. The two less successful groups had higher score in desire for more education (P=0.0383) and their will for changing the life style (P=0.0467). Discussion: Our individually-tailored intervention lowered the decisional conflict of women on their necessary lifestyle changes as well as their CVD risks. However, the less successful women neither lost their hope nor desire for help. Conclusion: The study indicates that individual women's characteristics which affect the compliance with intervention should be studied in detail, and utilized to additionally modify the interventions to more significant reduction of the CVD risk. Acknowledgements: We thank Dr. Sandor G. Vari, MD, Director of the International Research and Innovation in Medicine Program, Cedars – Sinai Medical Center, Los Angeles, CA, USA and President of the RECOOP HST Association for the initiation of the project and helpful comments. We also thank Dr. Ian Pitha, MD, PhD, Head of Laboratory for Atherosclerosis Research, Centre for Experimental Research, Institute for Clinical and Experimental Medicine (IKEM), Prague, Czech Republic for the critical review of our research plan

Electronic Interactive Poster Presentation Influence of acute and intermittent hyperbaric oxygenation on the reactivity of the blood vessels in healthy Sprague-Dawley rats Mihaljević, Z.1, Drenjančević, I.1

1Department of Physiology and Immunology, Faculty of Medicine Josip Juraj Strossmayer, University of Osijek, Croatia.

Corresponding author: ines.drenjancevic@mefos.hr

Key words: rat, hyperbaric oxygen therapy, vascular reactivity, oxidative stress Introduction: The effect of hyperbaric oxygenation (HBO2) on vascular function has been intensively studied in last decade and the understanding of the mechanisms of its action has been emerging, particularly in the diabetes mellitus. Our results suggest that repeated exposure to hyperbaric oxygen and the time between the two therapies can be viewed as intermittent pseudohypoxia and it is possible to trigger other adaptive mechanisms than previously observed. Oxygen partial pressure changes may lead to changes in the synthesis of metabolites of arachidonic acid (AA). This preliminary study aimed to examine the HBO2 effects on the vascular reactivity to acetylcholine (ACh) and hypoxia in aortic ring of healthy rats. Methods: Healthy Sprague-Dawley 9-12 weeks old rats were divided in four experimental groups (1. healthy control; 2. acute HBO2 exposure; 3. 24 hour after acute exposure; and 4. 4 subsequent days of HBO2 exposure). Isolated aortic rings reactivity to ACh and hypoxia was studied on Isolated Organ Bath (Experimetria LTD). The indicators of oxidative stress were measured by TBARS and FRAP methods. Results: Acute exposure of healthy animals (group 2) to HBO2 leads to the impaired vasorelaxation response to both ACh and hypoxia, not present in the groups 1., 3. And 4. Discussion: Duration of HBO2exposure diversely affects the vascular reactivity to different stimuli. Conclusion: Results indicate increased oxidative stress in acute exposure to HBO2, lost in other HBO2 treatments. Acknowledgments: This is preclinical animal research, partially supported by grants of Ministry of Science, Education and Sports, Croatia, # 219-2160133-2034 and # 219-0000000-0328 . All experimental procedures conformed to the European Guidelines for the Care and Use of Laboratory Animals (directive 86/609) and were approved by the local Ethical Committee #2158/61-02-139/2-06.

Comprehensive analysis of ischemic conditioning techniques in a porcine model of acute myocardial infarction András Makkos medical student VI., Semmelweis University, Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary Corresponding author: Andras Makkos makkosandras@gmail.com Key words: schemic conditioning, Ichemic postconditioning, Remote ischemic conditioning, MRI, T2-oedema reduction Introduction: Ischemic pre-, post- and remote conditioning are cardioprotective in animal models of acute myocardial infarction (AMI). However, their effect in clinical trials is still inconclusive, which could be attributed to the difficulties of the evaluation of cardioprotection. Our aim was to compare the cardioprotective effect of ischemic post- and remote conditioning by histological staining and magnetic resonance imaging (MRI) after AMI in a clinically relevant porcine model. Methods: Animals were randomized into ischemic (Isch), pre- (IPreC), post- (IPostC) and remote conditioned (RIC) groups. In general anesthesia, the anterior descending branch of the left coronary artery (LAD) was occluded for 90 min with balloon catheter (index ischemia). In IPreC group before the index ischemia 3×5-5 min, in IPostC group after the index ischemia 6×30-30 sec LAD occlusion/reperfusion cycle was applied. RIPerC performed by 4×5 min compression of the hind limb. Results: After 3 hours of reperfusion, myocardial necrosis was significantly decreased by IPreC, but not by IPostC and RIC as assessed by histological staining (Isch: 38.12±6.05%, IPreC: 12.15±2.94%*, IPostC: 38.14±4.89%, RIPerC: 32.43±3.19%; % of AAR). There was no difference in the area at risk (AAR) among groups. As evaluated with cardiac MRI (late gadolinium enhancement) after 3 days, myocardial necrosis was not changed among groups (Isch: 14.34±2.39%, IPreC: 9.01±2.32%, IPostC: 12.47±2.0%, RIC: 14.33±1.75%; % of left ventricle), while myocardial edema (T2-weighted MRI) was significantly decreased by IPostC and RIC (Isch: 30.38±4.05%; IPostC: 18.52±1.80%*; RIPerC: 17.40±2.68%*; % of left ventricle). Microvascular obstruction was also decreased by IPreC and IPostC (Isch: 1.25±0.62%; IPreC: 0.12±0.12%*; IPostC: 0.26±0.15%*; % of left ventricle). *p

The Role of Oxidative Stress in Development of Impaired Vascular Response Under The Influence Of High Salt Intake in Sprague-Dawley Rats Cosic A, Drenjancevic Department of Physiology and Immunology, Faculty of Medicine University of J.J. Strossmayer in Osijek, Osijek, Croatia The corresponding author: Ines Drenjancevic; ines.drenjancevic@mefos.hr

Keywords: antioxidative enzymes, flow-induced dilation, high salt diet, middle cerebral artery, oxidative stress Introduction: This study aimed to determine the in vitro flow-induced dilatation (FID) mechanisms of middle cerebral artery (MCA), the expression of antioxidative genes, genes involved in dilatation pathways, level of leukocyte activation and the role of oxidative stress in Sprague-Dawley (SD) rats on a high salt (HS) diet. Methods: 11-weeks old healthy male SD rats were divided in low salt group (LS) given standars rat chow (0.4%NaCl) and HS group given food containing 4%NaCl for 1 week with/without superoxide dismutase (SOD) mimetic TEMPOL in vivo. MCA reactivity in response to FID in the absence/presence of inhibitors L-NAME, INDO, MS-PPOH and TEMPOL in vitro was examined. mRNA levels of enzymes from brain blood vessels were determined with rtPCR. Leukocyte activation and level of oxidative stress in leukocytes from blood and peripheral organs were mesured by flow cytometry. Blood pressure, lipid peroxidation products and antioxidative capacity were also measured. Results: FID was reduced in HS group compared to LS group. Inhibitors (independantly) significantly reduced FID in LS group. FID in HS group was mediated by NO. TEMPOL restored FID in HS group to basal level. Expression of GPx4 and iNOS in HS group was significantly decreased. Antioxidant enzymes activity and BP were not affected by HS diet. HS intake significantly increased basic reactive oxygen species (ROS) production in the leukocytes of mesenteric lymph nodes and splenocytes, and intracellular production after stimulation in peripheral lymph nodes. Discussion: Mechanisms of FID are different in LS and HS groups. Low GPx4 expression, increased superoxide production in leukocytes, together with decreased iNOS expression underlies increased oxidative stress and reduced NO bioavailability in HS group, which caused impairment of FID in HS group without changes in BP. Conclusion: FID in HS diet is determined by synthesis of ROS in the vascular wall and affects vascular reactivity mechanisms. Sources of Funding: This work has been supported in part by Croatian Science Foundation under the project # IP-2014-09-6380 and in part with VIF-MEFOS-15 (Faculty of Medicine Osijek, Croatia). Acknowledgements: I am grateful to my mentor professor Ines Drenjancevic and to all colleagues from the Department of Physiology and Immunology for helpful advices and assistance during the development of the study. Ethical Approval: All experimental procedures are conformed to the European Guidelines for the Care and Use of Laboratory Animals (directive 86/609). They were approved by the local Ethical Committee (Faculty of Medicine, University of Osijek; Class: 602-04/14-08/06; No.: 2158-61-07-14-04) and Ministry of Agriculture, Croatia (HR-POK-005).

Electronic Interactive Poster Presentation- Neurodegenerative Diseases

Evaluation of putative cannabinoid ligands as modulators of neurodegeneration Senkiv J.V.1, Golota S.M.2, Irving J.A.3, Lesyk R.B.2, Stoika R.S.1 1. Department of Regulation Cell Proliferation and Apoptosis, Institute of Cell Biology NAS of Ukraine, Lviv, Ukraine 2. Department of Pharmacy, Organic and Bioorganic Chemistry, Danylo Halycky Lviv National Medical University, Lviv, Ukraine 3. School of Biomedical and Biomolecular Science, University College of Dublin, Dublin, Ireland Corresponding author Yulia Shenkiv Yu.senkiv@gmail.com

Key words (max 5): G-protein-coupled receptor (GPCR), putative cannabinoid ligands, neuronal function Introduction. Alzheimer's disease is a complex and progressive neurodegenerative disorder. The available therapy is limited to the symptomatic treatment and its efficacy remains unsatisfactory. The G-protein-coupled receptor (GPCR) superfamily of integral proteins is the biggest family of signal transducers, comprised of 1000 members. Considering their prevalence and functional importance, it is not surprising that 60 % of drugs target GPCRs. Cannabinoid ligands have neuroprotective actions and may have utility in the treatment of neurodegeneration. The major CNS targets for cannabinoids include the classical CB1 cannabinoid receptor and also GPR55. Although GPR55 is potently activated by the endogenous lysophospholipid, L-alysophosphatidylinositol (LPI), it is sensitive to a number of synthetic cannabinoid ligands. Methods. In this study, we used a range of functional assays to compare the pharmacological activity of selected cannabinoid ligands that were synthesized by the team of prof. R. Lesyk. These are: Les-3105, Les-2769, Les-2615, Les-2659, Les-3836, and LPI (positive control). The compounds were evaluated in cultured rat embryonic cortical neurons and immortalized cells of HEK293 line engineered to stably express recombinant human GPR55. Stimulation of protein kinase (ERK1/2) mitogen activated kinase MAP-kinases and of cAMP response element binding protein (CREB) were measured by Western-blot analysis, while CREB activation was additionally monitored using confocal imaging of nuclear phospho-CREB labelling. Results and Discussion. Striking effects of the compounds on pCREB activation in neurons were observed. Current studies are directed at evaluating their effects on GPR55 and CB1 activity. Conclusion. We have evaluated a new series of cannabinoid ligands which have clear effects on neuronal function that might be beneficial with respect to neurodegeneration. In particular, activation of CREB by phosphorylation has been implicated in cell survival. The molecular precise targets for these ligands are currently under evaluation. Acknowledgements. We thank the Physiological Society of the UK for travel grant awarded to Julia Senkiv. All animal experiments were conducted in accordance with S.I. No. 543/2012 - European Union (Protection of Animals Used for Scientific Purposes) Regulations 2012. This Irish legislation serves the purpose of giving effect to the European Union Directive 2010/63/EU. The structure of this study and the experimental procedures conducted were approved by the Health Products Regulatory Authority (HPRA)

Abstracts for Nanobiomedicine Research ORAL PRESENTATIONS

Research activity at Institute of Cell Biology, NAS of Ukraine: achievements, problems and perspectives of development within RECOOP-HST Association. Stoika R.S. Department of Regulation of Cell Proliferation and Apoptosis, Institute of Cell Biology (ICB), NAS of Ukraine, Drahomanov Str. 14/16, Lviv, 79005, Ukraine Corresponding author: Rostyslav Stoika stoika@cellbiol.lviv.ua In 2015-2016, the following projects have been executed by our team in collaboration with RECOOP and none-RECOOP partners. NanoBioTech research projects: 1) Imaging of Formalin-Fixed, Paraffin-Embedded melanoma tissues using specific lectin-functionalized nanocrystals. Since 2016, this project is performed at Lviv National Medical University. Future studies will be focused on detection of functionalized nanocrystals using X-ray excitation for X-ray compatible detection systems [10,11,12]. 2) Biomedical application of novel polymeric particles of nano- and micro-scale with a reactive shell suitable for bio-functionalization (in collaboration with Lviv National Polytechnic University) [14]. Enhanced anticancer activity and circumvention of drug resistance mechanisms was demonstrated at applying these polymeric/phospholipidic nanocarriers. 3) Targeting drug-resistant tumor cells in vitro and drug-resistant tumor in vivo by Fullerene C60/anticancer drug nanoconjugates [6, 15]. 4) Novel oligoelectrolyte-based non-viral gene delivery systems for genetic transformation of eukaryotic (yeast, plant, mammalian) cells [1,9