Program and Proceedings

International Symposium on Neuroscience and Biological Psychiatry

"STRESS AND BEHAVIOR: YEREVAN-2013"

September 20-21, 2013 Yerevan, Armenia

In conjunction with the Jubilee Conference of Orbeli Institute (September 22-24, 2013)

In partnership with:
L.A. Orbeli Institute of Physiology, National Academy of Sciences, Armenia

Metris B.V., Netherlands

Metris IT Solutions LTD, Armenia

Institute of Experimental Medicine, Russian Academy of Medical Sciences, Russia

U S BP

Ukrainian Society for Biological Psychiatry, Ukraine

FINAL PROGRAM DAY 1, FRIDAY, 20 SEPTEMBER 2013

8.30 9.30 9.50 Registration OFFICIAL OPENING AND WELCOMING (Ayvazyan N, Klimenko VM, Kalueff AV, Bachdasarian L) PLENARY LECTURE 1: NEUROBIOLOGICAL BASIS AND COMPENSATORY MECHANISMS IN PTSD. Klimenko VM, Tsikunov SG, Research Institute of Experimental Medicine NWB RAMS, St. Petersburg, Russia PLENARY LECTURE 2: STRESS AND NEURODEGENERATION. Aghajanov MI, Yenkoyan KB, Department of Biochemistry, Heratsi Yerevan State Medical University, Yerevan, Armenia Coffee break SESSION I: CLINICAL ASPECTS OF PTSD, STRESS AND BEHAVIOR Chairs: Klimenko VM, Tadevosyan AS 11.35 MEMORY MY EVIL LORD. Tadevosyan AS, Department of Stressology of Armenian Medical Institute (AMI), Armenian Association of Biological Psychiatry WFSBP, Stress Center MLSA RA, Yerevan, Armenia 11.50 PRACTICAL STRESSOLOGY. BORDERLINE MEDICINE (PRESENTATION OF THE MONOGRAPH). Tadevosyan AS, Stress Center MLSA RA, Yerevan, Armenia 12.00 INFLAMMATORY MEDIATORS IN THE BLOOD OF KARABAKH COMBAT VETERANS WITH POSTTRAUMATIC STRESS DISORDER. Hovhannisyan L, Mkrtchyan G, Tadevosyan M, Sukiasyan S, Boyajyan A, Institute of Molecular Biology NAS RA, Armenian Society of Biological Psychiatry WFSBP, Stress Center MLSA RA, Yerevan, Armenia 12.15 ANALYSIS OF GENE EXPRESSION IN POSTTRAUMATIC STRESS DISORDER BY APPLICATION OF BIOINFORMATICS TOOLS. Arakelyan A, Nersisyan L, Aslanyan L, Insitute of Molecular Biology NAS RA, Institute for Informatics and Automation Problems NAS RA, Armenian Society of Biological Psychiatry WFSBP, Yerevan, Armenia 12.30 EFFECTIVENESS OF PHYTOTHERAPY AFTER PERIPHERAL NERVE INJURY. Gevorgyan LR, Chavushyan VA, Simonyan KV, Orbeli Institute of Physiology, Yerevan, Armenia 12.45 THE ROLE OF BRAIN INJURY IN PTSD WITH PSEUDOSEIZURES: A CASE STUDY. Margaryan SP, Armenian Society of Biological Psychiatry WFSBP, Stress Center MLSA RA, Yerevan, Armenia 13.00 HETEROGENEITY AMONG PTSD PATIENTS: TOWARDS UNDERSTANDING A DISSOCIATIVE SUBTYPE OF PTSD. Zaba M, Gall-Kleebach D, Schubert C, Schmidt U, MaxPlanck-Institute of Psychiatry, Munich, Germany 13.15 SOME LIFELONG SOCIAL CHARACTERISTICS OF EX-COMBATANTS OF KARABAKH WAR WITH POST-TRAUMATIC STRESS DISORDER. Margaryan SP, Sukiasyan SH, Armenian Society of Biological Psychiatry WFSBP, Stress Center MLSA RA, Yerevan, Armenia 13.30 NEW VIBROACOUSTIC SENSOR BASED ON THE SFCO TECHNOLOGY: ITS APPLICATION IN BIOMEDICAL INVESTIGATION. Khachunts AS, Gevorgyan SG, Muradyan ST, Gevorgyan VS, Institute of Physiology NAS, Precision Sensors/Instrumentation D&P Ltd. PSI, Faculty of Physics, Yerevan State University, Yerevan, Armenia 13.45 HISTOLOGICAL PECULARITIES OF HUMAN PLACENTA AFTER CHRONIC PSYCHO-EMOTIONAL STRESS. Aznauryan AV, Khudaverdyan AD, Heratci Yerevan State Medical University, Yerevan, Armenia 14.00 POSTER PRESENTATION: DEPRESSION AMONG THE ROMANIAN ADULTS. Abram Z, Department of Hygiene, University of Medicine and Pharmacy, Tirgu-Mures, Romania Lunch Break PLENARY PRESENTATION 3: FEASIBILITY STUDY OF FEAR CONDITIONING PROTOCOL (Laboras/Sonotrack presentation) Bachdasarian L, Metris B.V., Netherlands

10.30 11.10 11.35

14.15 15.15

16.00

SESSION II: PRE-CLINICAL ASPECTS OF PTSD, STRESS AND BEHAVIOR Chairs: Aghajanov MI, Bulthuis R 16.00 CHANGES IN HIPPOCAMPUS AND CEREBELLUM DEPENDENT BEHAVIOR OF RATS IN HYPOKINETIC CONDITIONS. Navasardyan GA, Yeritsyan NB, Safaryan KS, Grigoryan AS, Yerevan State Medical University, Yerevan, Armenia 16.15 ACTH AND CORTISOL LEVELS IN BLOOD OF PREGNANT RATS AND THEIR OFFSPRING IN THE ACTION OF CHRONIC STRESS. Khudaverdyan DN, Saroyan MY, Khudaverdyan AD, Ghambaryan HK, Heratci Yerevan State Medical University, Yerevan, Armenia 16.30 EXPERIMENTAL ANIMAL MODELS OF PTSD: UPDATE AND FUTURE CHALLENGES. Kalueff AV, Stewart AM, ZENEREI Institute, Slidell, USA 16.45 POSTER PRESENTATIONS: BAROREFLEX SYSTEM AS A TARGET OF INTERVENTION OF THE ORIGINAL VEGETOTROPIC DRUGS AND THE OBJECT OF MATHEMATICAL MODELING. Shirinyan ME, Shirinyan EA, Mnjoyan Institute of Fine Organic Chemistry (IFOC) STCOPC NAS RA, Yerevan, Armenia ENHANCING CAPABILITIES FOR STUDY MEMORY AND LEARNING IN THE SHUTTLE BOX AVOIDANCE TASK. Berezhnoy DS, Inozemtsev AN, Department of Physiology of Higher Nervous Activity, Lomonosov Moscow State University, Moscow, Russia POSTER AND DEMONSTRATION (LABORAS, SONOTRACK, DSI) THE NEED FOR MULTI-MODAL AND MULTI-FUNCTIONAL MEASUREMENT SYSTEMS IN LABORATORY ANIMAL RESEARCH AND MATRIX ANALYSIS I. Bachdasarian L, Bulthuis R, Molenwijk E, Karkishenko V, Revyakin A, Metris B.V., Hoofddorp, Netherlands, Scientific Centre Bio-Medical Technology Moscow, Russia POSTER PRESENTATION: STRESS-FREE MONITORING OF DEVELOPMENT USING THE LITTER [DAM+PUPS] AS MEASURING UNIT. A PERINATAL STUDY IN RATS WITH METHYLMERCURY. de Groot D, Slieker R, Bulthuis R, Bachdasarian L,TNO Zeist, Zeist, Metris BV, Hoofddorp, Leiden University Medical Center, Leiden, Netherlands

17.00

DAY 2, SATURDAY, 21 SEPTEMBER 2013

9.00 VISIT TO THE GENOCIDE MUSEUM YEREVAN WHO ARE THEY? ARMENIAN WOMENS EXPERIENCE DURING AND AFTER THE GENOCIDE. Aleksanyan A, Armenian Genocide Museum-Institute, Yerevan, Armenia IMPLICATIONS OF GENOCIDE DENIAL ON THE VICTIM GROUP: THE ARMENIAN CASE. Gzoyan E, International Relations, LLM, Yerevan, Armenia

12:00 Lunch Break 13.00 SESSION III: MOLECULAR-GENETIC ASPECTS OF COGNITION, PHYSIOLOGY AND STRESS Chairs: Kalueff AV, Sargisyan S, Strekalova T 13.00 COMPARATIVE CHARACTERISTICS OF DIFFERENT SUBPOPULATIONS OF THE IMMUNE COMPLEXES IN PSYCHO-NEUROLOGICAL DISORDERS: IMPLICATION TO DISEASE-RELATED MOLECULAR PATHOMECHANISMS. Chavushyan A, Hovhannisyan L, Mkrtchyan G, Hovsepyan M, Arakelyan A, Hakobyan S, Khoyetsyan A, Petrosyan M, Harutyunyan K, Musaelian R, Ayvazyan V, Hovhannisyan G, Aroutiounian R, Boyajyan A, Institute of Molecular Biology NAS RA, Armenian Society of Biological Psychiatry WFSBP, rtsakh Scientific Center of Nagorno-Karabakh, Yerevan State University, Yerevan, Armenia 13.15 DISC1 REGULATION OF D-SERINE PRODUCTION IS SPECIFIC TO ASTROCYTES. Abazyan S, Ma TM, Abazyan B, Nomura J, Yang C, Seshadri S, Sawa A, Snyder SH, Pletnikov MV, Department of Psychiatry and Behavioral Sciences, The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA, Division of Molecular Medical Science, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan 13.30 EFFECT OF ELECTRIC FOOT SHOCK STRESS ON TRYPTOPHAN CONCENTRATION IN PLASMA AND RED BLOOD CELLS OF DIFFERENT AGE RATS.

Nevoia A, Ciochina V, Garaeva S, Postolati G, Redcozubova G, Institute of Physiology and Sanocreatology MAS, Chisinau, Moldova 13.45 MOLECULAR AND GENETIC INDICATORS OF APOPTOTIC AND SYNAPTIC PLASTICITY ALTERATIONS IN POSTTRAUMATIC STRESS DISORDER AND SCHIZOPHRENIA. Boyajyan A, Avetyan D, Atshemyan S, Sukiasyan S, Gevorgyan A, Mkrtchyan G, Zakharyan R, Chavushyan A, Melkumova M, Torosyan S, Institute of Molecular Biology NAS RA; Armenian Society of Biological Psychiatry WFSBP, Russian-Armenian Slavonic University, Stress Center MLSA RA, Psychiatric Medical Center MH RA, Yerevan, Armenia ROUND TABLE: PTSD (POST-TRAUMATIC STRESS DISORDER) AND GENETICS: HOW GENOCIDE AND WAR INFLUENCE GENETICS AND BEHAVIOR Chairs: Bachdasarian L, Tadevosyan AS, Margaryan S PTSD AND SIGNALS EFECTS ON HUMANS AND ANIMALS. Bachdasarian L, Metris BV, Hoofddorp, Netherlands

14.00

15:00 Coffee break 15:20 PRODUCT PRESENTATION (METRIS AND DSI) THE NEED FOR MULTI-MODAL AND MULTI-FUNCTIONAL MEASUREMENT SYSTEMS IN LABORATORY ANIMAL RESEARCH AND MATRIX ANALYSIS II. Bachdasarian L, Bulthuis R, Molenwijk E, Gaburro S, Meuth P, Metris B.V., Netherlands, Data Sciences International, St. Paul, USA, Department of Neurology, Inflammatory Disorders of the Nervous System and Neurooncology, Institute of Physiology I, Westflische Wilhelms-Universitt Mnster, Germany SESSION IV: NEUROBIOLOGY OF STRESS IMPACT ON BRAIN PLASTICITY AND MEMORY. Chair: Yeritsyan NB 16.00 THE IMPACT OF EARLY PHARMACOLOGICAL TRAUMA ON BRAIN ANATOMY IN GERBILS. Lehmann K, Institute for General Zoology and Animal Physiology, Friedrich-Schiller University Jena, Jena, Germany 16.30 SPECIAL LECTURE III: STRESS, EARLY LIFE EVENTS AND MEMORY FORMATION. Krugers H, University of Amsterdam, Swammerdam Institute for Life Sciences, Amsterdam, Netherlands 17.10 ENVIRONMENTAL FACTORS TO INDUCE OR TO RELEAVE STRESS AND DEPRESSIVE TRAITS: A STUDY ON MICE. Anthony DC, Chernopyatko AS, Strekalova T, Department of Pharmacology, University of Oxford, Oxford, UK, Timantti AB, Stockholm, Sweden, Department of Neuroscience, School for Mental Health and Neuroscience, Maastricht University, Maastricht, Netherlands 17.40 LEARNING-ASSOCIATED STRESS: PLASTICITY UNDERLYING MEMORY. Yeritsyan N, Department of Neurophysiology, Leibniz Institute for Neurobiology, Magdeburg, Germany

16.00

18:10 OFFICIAL CLOSING CEREMONY. ANNOUNCING FUTURE ISBS CONFERENCES

SYMPOSIUM VENUE
Conference Hall of the L.A. Orbeli Institute of Physiology of the Armenian National Academy of Sciences (Yerevan) the leading biomedical research institute of Armenia. The L.A. Orbeli Institute of Physiology was founded in 1943, and named after Prof. Leon A. Orbeli, a prominent neuroscientist and Pavlovs friend and follower. Currently, the Institute is a leading center of neuroscience research in Armenia, and a regular host of various scientific forums and national and international neuroscience meetings. Address: Tel.: URL: 22, Orbeli Bros. St.,Yerevan 0028, Armenia (+374 10) 272247 www.physiol.sci.am www.neuroscience.am/orbeli.php

SYMPOSIUM ORGANIZERS
International Organizing Committee: Symposium Co-Chairs: Allan V. Kalueff, PhD (New Orleans, USA), Director, ZENEREI Institute; President of ISBS and USBP (National Society of WFSBP) Victor M. Klimenko MD, PhD (St-Petersburg, Russia), Head, Pavlov Department of Physiology, Institute of Experimental Medicine Ronald Bulthuis (Hoofddorp, Netherlands), Director, Metris B.V. Levon Bachdasarian, PhD (Hoofddorp, Netherlands), Manager, Metris B.V. Scientific Program Committee: Chair: Victor Klimenko MD, PhD (St-Petersburg, Russia) Members: Naira Ayvazyan, PhD (Director, Orbeli Institute of Physiology, Yerevan, Armenia) Allan Kalueff, PhD (Director, ZENEREI Institute, New Orleans, USA) Armen Sargisyan, PhD (Sr. Researcher, Orbeli Institute of Physiology, Yerevan, Armenia) Ada Tadevosyan, MD, PhD (Founder and Hon. Director, Stress Center, Armenian Society for Biological Psychiatry WFSBP, Yerevan, Armenia) Mikhail Aghajanov, MD (Chair of Biochemistry, National Medical University, Yerevan, Armenia) Ronald Bulthuis (Director, Metris B.V., Hoofddorp, Netherlands) Levon Bachdasarian, PhD (Manager, Metris B.V., Hoofddorp, Netherlands) Samvel Margaryan, MD, PhD (Stress Center MLSA, ISTSS/WFSBP, Yerevan, Armenia) Local Organizing Committee Co-Chairs: Ada Tadevosyan, MD, PhD (Founder and Hon. Director, Stress Center, Armenian Society for Biological Psychiatry WFSBP,Yerevan, Armenia) Armen Sargisyan, PhD (Sr. Researcher, Orbeli Institute of Physiology, Yerevan, Armenia) Symposium Official Travel Agent: Levon Travel Tours, LTD (Yerevan, Armenia) ISBS Secretary General: Nataliya A. Zinevych, PE (New Orleans, LA, USA)

SYMPOSIUM ABSTRACTS
DAY 1, FRIDAY, 20 SEPTEMBER 2013
PLENARY LECTURE 1: NEUROBIOLOGICAL BASIS AND COMPENSATORY MECHANISMS IN PTSD. Klimenko VM, Tsikunov SG, Research Institute of Experimental Medicine NWB RAMS, St. Petersburg, Russia INTRODUCTION: Study of the consequences of severe psychic trauma are highly relevant in the current conditions of natural disasters and regional conflicts. Advancing the treatment and prevention of disorders caused by the vital stress is often difficult because of the problem with creating valid animal models of human mental illnesses. METHODS: We have developed original models of psychic trauma (PT) and post-traumatic stress disorder (PTSD) in rats, close to the same pathology in humans. Their essence consists in the emotional experience of groups of animals of the death of one of the members of their community from the actions of a predator snake (python). We performed a complete validation of the developed models and showed their compliance with all major criteria of modeling - the etiology, symptoms, neurochemical abnormalities and sensitivity to pharmacological agents. RESULTS AND DISCUSSION: Using these models, we obtained first data on the reaction the brain and other body systems to extreme mental stressors. PT in rats causes long-lasting behavioral disturbances, which can be considered as a model of PTSD. In rat plasma at different times after vital stress, we detected variations in the lipid metabolism, manifesting changes in the spectrum of plasma atherogenic lipids. We noted morphological changes in the adrenal glands, indicating their hypofunction. In the brain, we found selective increases in lipid peroxidation of biological membranes in the hippocampus and cerebral cortex. These data reflect the increased formation of free radicals and development of oxidative stress, which may have a damaging effect on the membrane of cells. We further revealed increased serotonin turnover in the midbrain and striatum. We also showed that the experience of a single PT with a real threat to life, leads to long-lasting structural and functional abnormalities in the brain. After the acute PT, we detected more diffuse dying nerve cells in the hippocampus, hypothalamus and cerebral cortex. We also found novel, not previously described, disturbances of immune-histochemical reaction to nuclear protein NeuN of the nerve cells in the hippocampus and limbic cortex. We further revealed inhibition of proliferation from progenitor stem cells in the subgranular layer hippocampal dentate gyrus and in the subventricular zone of the lateral ventricles of the brain, which indicates that processes of neurogenesis are involved in mechanisms of development and compensation of PTSD. We experimentally demonstrated the possibility of correction of deficits by means of targeted regulation of the activity of neurotransmitter and neuropeptide systems of the brain. Overall, the observed abnormalities may be new, previously unknown targets, for the impact of drugs, such as synthetic analogs of endogenous neuropeptides. STRESS AND NEURODEGENERATION. Aghajanov MI, Yenkoyan KB, Department of Biochemistry, Heratsi Yerevan State Medical University, Yerevan, Armenia It has been known for decades that stress contributes to heart disease, insomnia, digestive disorders and a host of other health problems. On the one hand acute stress may be useful for brain plasticity, on the other, there is a mounting evidence that chronic ongoing stress may lead to pathological changes in stress circuitry and trigger the damage of brain neurons, which eventually results in neurodegeneration. One example of stress-induced neurodegenerative events is Alzheimers Disease (AD). The objective of the present study was to focus on principal mechanisms of neuronal damage and survival. The modeling of neurodegeneration was performed by intracerebroventricular injection of different fragments of beta-amyloid. The complex of proteoglycans of embryonal genesis (PEG) created by L. Mkrtchyan was used as a survival agent against stressful stimuli. The role of pivotal systems involved in neuronal protection pathways like IGF-1, NGF, neuroactive amino acids (aspartate, glutamate, glycine, taurine, GABA) was analyzed in details. In context of monoaminergic disinhibition phenomena the measurement of monoamines and their derivates (epinephrine, norepinephrine, homovanillic acid, 5hydroxyindolacetate) in brain was performed, together with oxidative stress markers (spontaneous and induced chemiluminescence). Concurrently, morphological (hematoxylin-eosin staining, histochemistry), immunohistochemical, electrophysiological (evoked tetanic and posttetanic activity) and behavioral (elevated plus-maze, Y-maze) analyses were performed, which provide essential characteristics of neuronal damage and survival. Special emphasis was made on studying the process of neurogenesis in specific regions of adult brain. The investigation of different markers of neuronal stem cells proliferation and differentiation (Nestin, Vimentin, GFAP, NeuN, NSE, Wnt, VEGF, Sox2, Sox10, Sox14) and BrdU assay gave a unique possibility to deeply understand the mechanisms of new neurons formation in adult brain in vivo. Remarkably, the proteoglycans of embryonal genesis (PEG) activated stem-cell proliferation and further differentiation in adult brain. In conclusion, we provide a strong evidence that

PEG may be considered as an anti-stress agent able to reduce the effects of stress on neurons, and consequently prevent their damage. SESSION I: CLINICAL ASPECTS OF PTSD, STRESS AND BEHAVIOR Chairs: Klimenko VM, Tadevosyan AS MY MEMORY - MY EVIL LORD. Tadevosyan AS, Department of Stressology of Armenian Medical Institute (AMI), Armenian Association of Biological Psychiatry WFSBP, Stress Center MLSA RA, Yerevan, Armenia INTRODUCTION: The memory system should be considered as the basis of a person's life in the complex human-environment open system, controlled by consciousness and will. Due to the peculiarities of this memory human being is the only one in the world with the capacity of succession of times past to the present and predicting the future. Time links are connected into a single life destiny and into its whole living by individual, nation, humanity. Network of memory is a hierarchy of levels, registers, different mechanisms of information storage, the options of interaction and interdependence and the inverse effect on the fate. OBJECTIVE: Investigation of psychopathology of memory in individuals with different types of PTSD. METHODS: Clinical material gathered for > 30 years: outpatients with traumatic stress disorder of different genesis and inpatients of the Center Stress, as well. RESULTS AND DISCUSSION: The moment of trauma obtaining is an act, which has a clear spatiotemporal boundaries. Depending on the type of traumatic memory it is forming traumatic constellation having specific characteristics. Further they determine the subsequent vector of persons behavior and development. The echo-effects (conventional definition) have a mental holographic effect a traumatic geschtalt. Each echo-effect has its own mechanism of formation, but all they are polysensory and include both affective and behavioral components of the moment of trauma. Sensory echo-effect (flash back) - a mirror image of the event, probably due to the activation of mirror neurons caused by neurobiological mechanism of the first phase of stress response. In cases of cognitive echoeffect, the leading role belongs to thinking, which leads to chronic stress resulting from the second phase of the GAS with the effect of the conversion to an organ. This effect is capable to lead from functional disturbances to structural changes in the organ-target (Tadevosyan, 2002, 2003, 2011). The effects of traumatic memory can be conscious and unconscious. Clinical cases of different traumatic memory will be presented. CONCLUSION: The type of traumatic memory has prognostic importance, defining the risk of development of some symptoms of PTSD, the option of psychosomatic disease, the possibility of inheriting of traumatic perception of the world in the future and of appropriate formation of the personality in both directions of destruction and growth. PRACTICAL STRESSOLOGY. BORDERLINE MEDICINE. PRESENTATION OF THE MONOGRAPH. Tadevosyan AS, Stress Center MLSA RA, Yerevan, Armenia The proposed monograph is based on the extensive clinical observations as well as deep scientific analysis of the theory development of post-stress disorders. It was developed as an attempt to unify multiple approaches for the purpose of understanding of the problem. Currently, there is no comprehensive or broadly accepted approach to the ethnology of the PTSD. We choose the systematic adaptive approach as a method. The main theoretical outline of the monograph is based on the specificity of human adaptation. The author uses the concepts of the system of biological adaptation (passive adaptation) and the system of psychic adaptation (active adaptation), which comprises the mechanisms of the conscious self-regulation. The necessity of new adaptive mechanisms in human evolution based on the progressive development of consciousness, speech, and imagination, specificity of the memory system and expansion of stressful factors. The necessity to give meaning to everything and therefore to give affective response by mechanism of stress genesis (three-phase stress-response by H. Selye) resulted in "the expansion of biological possibilities of stress response." With relatively inflexible limitations of biological adaptation its expansion occurs by the mechanism of dissociation, which causes disintegration of the united stressor process (GAS by Selye). Its constituents (phases of anxiety, tension and asthenia) have acquired evolutionary functional autonomy. The conclusion about stress phase-oriented model of the disease inevitably arises. This model includes the whole diversity of anxious, depressive, psychosomatic and co-morbid disorders. The material is presented as a comparison of achievements of neurophysiology, psychoanalysis and psychopathology. This approach allows to offer to the physician a single concept of "human in medicine" and to develop pathogenesis tactics of combined therapy. This monograph is the continuation of Stressology" (2002), translated into English ("Man under Stress", 2003). Practical and theoretical frameworks outlined in the book are of practical importance in the development of Stressology as a form of preventive medicine. INFLAMMATORY MEDIATORS IN THE BLOOD OF KARABAKH COMBAT VETERANS WITH POST-TRAUMATIC STRESS DISORDER. Hovhannisyan L, Mkrtchyan G, Tadevosyan M, Sukiasyan S, Boyajyan A, Institute of Molecular Biology NAS RA, Armenian Society of Biological Psychiatry WFSBP, Stress Center MLSA RA, Yerevan, Armenia

INTRODUCTION: Development of evidence-based methods for treatment and prevention of posttraumatic stress disorder (PTSD; DSM-IV-TR code: 309.81) and disease-associated complications is an important healthcare, social and economic problem worldwide. In spite of many publications related to the risk factors, prevalence, clinical psychopathological symptoms and progression of PTSD, molecular pathogenesis of this disease is not clear yet. Promising findings indicate that the immune system alterations, particularly low-grade inflammatory reactions, exacerbate the course of PTSD and cause complications. The molecular mechanisms of the above mentioned alterations and their relation to the clinical psychopathological criteria of PTSD are not defined yet, which in a sufficient degree is limiting the progress in development of efficient measures of therapy and prevention of complications for this disease. OBJECTIVES: The aim of this study was to investigate the molecular mechanisms of immune system alterations in PTSD and clarify their role in the disease pathogenesis. To achieve this goal we assessed functional activities of the immune response mediators, the complement system and cytokine network, in PTSD and evaluate their potential association with psychopathological symptoms of PTSD. SUBJECTS AND METHODS: Study population includes 120 PTSD patients (Karabakh war veterans) and the equal number of age- and sex-matched healthy controls. The study was approved by the Ethical Committee of the Institute of Molecular Biology NAS RA (IRB 00004079). The enzyme-linked immunosorbent and hemolytic assays were used in this study. Data analysis was based on a broad range of methods of parametric and non-parametric statistics. Correlation of measured parameters with the expression of PTSD clinical symptoms was evaluated using Structured Clinical Interview for the DSM-IV Axis I Disorders (PTSD module) and Clinician-Administered PTSD Scale (CAPS). RESULTS: The results of this study demonstrated that PTSD is characterized by the disturbance of complement system functional activity, which is manifested at the level of hyperactivation of the classical and terminal complement pathways, deficiency of the complement key component C3 and hypo-activation of the complement alternative pathway. It was also shown that PTSD is characterized by increased levels of proinflammatory and chemotactic cytokines interleukin(IL)-1, IL-6, tumor necrosis factor-, monocyte chemotactic protein-1, and IL-8. A positive correlation between the levels of interleukin-1 and interleukin-6 and degree of expression of clinically significant PTSD symptoms was detected. CONCLUSIONS: Our study provides further evidence on implication of the immune system alterations in the pathogenesis of PTSD. The results obtained demonstrated that the complement cascade as well as proinflammatory and chemotactic cytokines are implicated in low-grade systemic inflammatory reactions associated with PTSD. Based upon the results obtained we also concluded that proinflammatory cytokines IL-1 and IL-6 may be considered as new therapeutic targets for PTSD treatment. ANALYSIS OF GENE EXPRESSION IN POSTTRAUMATIC STRESS DISORDER BY APPLICATION OF BIOINFORMATICS TOOLS. Arakelyan A, Nersisyan L, Aslanyan L, Insitute of Molecular Biology NAS RA, Institute for Informatics and Automation Problems NAS RA, Armenian Society of Biological Psychiatry WFSBP, Yerevan, Armenia We evaluated the functional gene sets affected in peripheral blood mononuclear cells of patients with posttraumatic stress disorder (PTSD) at both early and advanced stages of the disease. Data sets were obtained from Gene Expression Omnibus repository. Principal component analysis was used for identification of key component explaining the gene expression variability between patients and controls. Functional gene sets deregulated in disease as well as in mouse model have been assessed using Gene Set Enrichment Analysis (GSEA). The data obtained here provide strong evidence on the involvement of the inflammatory, neuronal, cytokine/growth factor signaling, and nucleotide metabolism pathways in PTSD progression. In addition, we showed that PCA/GSEA is an efficient tool to discriminate phenotype related gene expression variability from other factors and to have more complex look into molecular mechanisms underlying pathological condition. EFFECTIVENESS OF PHYTOTHERAPY AFTER PERIPHERAL NERVE INJURY. Gevorgyan LR, Chavushyan VA, Simonyan KV, Orbeli Institute of Physiology, Yerevan, Armenia INTRODUCTION: The repairing and regeneration of peripheral nerves is a very complex cytological process, its mechanism is unclear so far, and thus results in the lack of specific and effectual therapy and medicament. The herbs and their effective components have their own inimitable predominance in promoting peripheral nerve regeneration, such as their multi-factorial, multi-target and multi-functional action, abundant source, inexpensive. Thymus serpyllum have antioxidant, anti-inflammatory, anticholinesterase and GABA-modulating activity. The aim of present study was to estimate the neuroprotective effectiveness of Thymus serpyllum after crush-injury of rats sciatic nerve (SN). METHODS: Experiments were carried out on male albino rats divided into 3 groups: 1) intact 2) subjected to unilateral crush injury to the SN (control) and 3) the same in conditions of administration of Thymus serpyllum ethyl extract daily for 3 weeks from the day after SN injury. After 30 days we have recorded extracellular spike activity of lateral vestibular nucleus (LVN) neurons, regulatory start and execution of motor activity under high frequency stimulation of injured SN distal part. Before and after i/m single administration of therapeutic doses of Thymus was recorded spike activity of neurons in the LVN.

Analysis of sensory (test reflex abduction) and motor (static sciatic index) indices of functional recovery after crush injury in dynamics of development of neurodegeneraton and regeneration and morphohistochemical study of injured site of SN were carried out. RESULTS AND DISCUSSION: Electrophysiological data by revealing of relevant to norm a balance of types and values of expressness of excitatory and inhibitory responses of LVN neurons provide evidence of a recovery of conducting through the region of injury and normal neurotransmitters status of LVN neurons in Thymus group. Slowing of background and an increase of post-stimulus spiking in neurons of LVN was recorded within 15-80 min of Thymus action. A comparative analysis of indices in intact and injured lower extremities revealed the recovery of motor and sensory function on the 30 day under action of Thymus. Thin myelinated fibers, which bind distal and proximal parts of injury exist on morphohistochemical slices. CONCLUSION: These findings suggest that that administration of Thymus serpyllum to the crushinjured rat may accelerate the functional recovery of CNS by increasing the rate of SN regeneration. The stimulating effect of Thymus in excitatory and inhibitory neurotransmitters system is an important component of neuronal plasticity and protection. THE ROLE OF BRAIN INJURY IN PTSD WITH PSEUDOSEIZURES: A CASE STUDY. Margaryan SP, Stress Center MLSA RA ISTSS, Armenian Society of Biological Psychiatry WFSBP, Yerevan, Armenia INTRODUCTION: The differential diagnosis of true epileptic versus psychogenic fits is difficult. The research to date suggests a) PTSD is important in the development of pseudo-seizures; b) brain injury increases the risk of PTSD. Very few data is available about the interrelations of brain injury, PTSD and pseudo seizures. METHOD: Single case study. RESULTS: Patient S, 41 years old. There was no relevant medical history. At age 28, he saw the death of his wife and son under the ruins of earthquake. He was reserved, but then became rather quiet, and he was married after 4 months. At age 31, he joined volunteer soldiers, was affected by severe psychogenic factors during the war. At age 34 he was wounded associated with serious brain contusion. After the return he developed PTSD, had a strong guilt for the loss of friend, saw clearly his face. He became explosive, at the height of headache he became aggressive, once he struck heavily his daughter. He had amnesia what had happened. The GP referred him to the Center of Epileptology, where they did not reveal epilepsy, and diagnosed PTSD. During the inpatient treatment he became quiet, but continued to have periodically seizures on the height of headaches, lasting 3-10 min and ending with amnesia and expressed fatigue. He did well only after the antidepressive treatment. CONCLUSION: PTSD and pseudo seizures not developed after severe personal misfortune without brain injury, but developed after war distresses and brain contusion. Traumatic brain injury + traumatic stress = pseudo epilepsy? Clearly, this problem warrants prospective longitudinal studies. HETEROGENEITY AMONG PTSD PATIENTS: TOWARDS UNDERSTANDING A DISSOCIATIVE SUBTYPE OF PTSD. Zaba M, Gall-Kleebach D, Schubert C, Schmidt U, Max-Planck-Institute of Psychiatry, Munich, Germany INTRODUCTION: The dissociative subtype of PTSD was proposed to constitute a distinct nosologic entity among trauma spectrum disorders. However, empirical findings on this are scarce and the phenomenon of dissociation is still poorly understood. METHODS: PTSD patients and age and sex matched healthy controls were assessed with clinical interviews and standardized questionnaires for evaluation of their stress coping strategies and dissociative symptoms. Additionally, in PTSD patients, coping with the traumatic event as well as modulation of their stress coping strategies and dissociative symptoms in the course of therapy were analyzed. RESULTS AND DISCUSSION: PTSD patients employed more maladaptive stress coping strategies and exhibited more dissociative symptoms in comparison to healthy controls. Interestingly, PTSD patients exhibiting a high intensity of dissociative symptoms had a higher incidence of avoidance symptoms and maladaptive coping strategies providing support for the existence of the dissociative subtype of PTSD. Additionally, preliminary evaluation of the development of stress coping and dissociative symptoms in the course of therapy suggests that PTSD treatment might influence stress coping strategies. RESEARCH SUPPORT: Max-Planck-Institute of Psychiatry in Munich, Germany. SOME LIFELONG SOCIAL CHARACTERISTICS OF EX-COMBATANTS OF KARABAKH WAR WITH POST-TRAUMATIC STRESS DISORDER. Margaryan SP, Sukiasyan SH, Stress Center MLSA RA ISTSS, Armenian Society of Biological Psychiatry WFSBP, Yerevan, Armenia OBJECTIVE: To find out the peculiarities of different social factors in Armenian ex-combatants with posttraumatic stress disorder to highlight the main directions of therapy and healing. METHOD: The data of 105 patients - former participants of combat actions in Karabakh were analyzed. Using specially designed questionnaires the psychiatric and social states of the mentioned patients have assessed. They also completed Hopkins Symptom Inventory (SCL-90). RESULTS: The posttraumatic stress disorder (PTSD) was diagnosed according to ICD-10 criterion. Practically all combatants had traumatic brain injuries (including contusions). In unfavorable social situation both organic and psychogenic factors caused respective changes of personality of ex-combatants. The average age of examined

10

patients was 44.6; of them 95 (90.48%) unemployed, moreover, practically all patients explain this fact mainly by their unsociable and aggressive nature. Practically nobody of other 10 working patients do not perform completely their professional duties they were just "tolerated at the workplace" since very often were consisted in the staff of their former more lucky combat comrade in arms. Herewith 28 (26.7%) patients did not have any group of disability, in spite of sufficiently long course of the disease (the average duration of disease is about 13.5 years). The course of their life and personality changes during the postwar years allows to suppose that hard social-economic conditions of life have receded into the background of moral motivations, having brought forth onward "the problems of the belly", in other words the motivations of ex-combatants inclined to compensation interrelations. 75 patients (71.43%) were married already before the war and continued to be married. Other 7 (6.7%) married during postwar years. 8 patients (7.62%) divorced, and 2 (1.9%) married again. 13 patients (12.4%) are not married. Patients in marriage and their family members, as well, noted extremely hard, unbearable, traumatizing character of relations in the family. For the whole period of stay in hospital (8-32 days) practically in half of the cases wives of patients never visited them in the hospital. CONCLUSION: The obtained data highlight the problems of therapy and healing of former war participants, which besides doubly medical, includes also social and humanistic aspects. NEW VIBROACOUSTIC SENSOR BASED ON THE SFCO TECHNOLOGY: ITS APPLICATION IN BIOMEDICAL INVESTIGATION. Khachunts AS, Gevorgyan SG, Muradyan ST, Gevorgyan VS, Institute of Physiology NAS, Precision Sensors/Instrumentation D&P Ltd. PSI, Faculty of Physics, Yerevan State University, Yerevan, Armenia INTRODUCTION: The progress in science, causing breakthrough changes in cutting edge technologies, is to a large extent formed by the use of the same technological innovations. METHODS: The creation of sensors based on the Single-layer Flat-Coil-Oscillators, activated by a low-power (backward) tunnel diode (SFCO-technology) is not an exception as well. The pioneering results are obtained in a basic superconductivity on the base of the highly sensitive this technology, and the new generation seismic detector is created, as well. Actually, the SFCO sensors with nano-metric level resolution can be classified into three groups. The absolute position and vibration sensors (the seismic sensors), the pressure sensors (the differential pressure gauges) and the vibro-acoustic sensors (the microphones), with a very large frequency band of sensitive operation ranging, theoretically, from 0 hertz (infrasound) up to 20 kHz. For all the three abovementioned groups detection is based on the frequency shift of the measuring SFCO oscillator. The precision of the measurements at the use of such sensors is determined by the 1-2 Hz stability of the oscillator base frequency F~20 MHz. Even negligibly small changes in the level of vibration and/or sound pressure result in a significant shift in a frequency of the SFCO oscillator measuring. RESULTS AND DISCUSSION: The feasibility of the use of the vibroacoustic SFCO sensor in medical and biological investigations is examined. The experiments were conducted on healthy volunteers from the laboratory staff (6 individuals, aged 27 to 59 years old). Vibroacoustic effects were recorded from the left carotid artery for the following states: relaxed, temporary voluntary breathing pause, and during Latin (Roman) alphabet letters pronunciation. The registered information, in general, is well correlated with the classical curves of sphygmogram waveform, however it contains much more sets of subcomponents. The spectral analysis of signals shows distinct peaks at the frequencies of breathing (~0.3 Hz) and Heart Rates (~1.2 Hz). In addition to main harmonics, we found 3-4 sub-harmonics of HR (~2.4, 3.6 and 4.8 Hz, respectively). The pause of breathing not only causes the respiratory component to disappear, but also modifies sub-harmonics of the HR. During the pronunciation of the letters on the recorded curves and on the corresponding spectra the ingredients of the specific pronounceable letter are observed (and their harmonics on the corresponding spectra, as well). Thus, the above-mentioned results provide sufficient evidence to conclude that the new vibro-acoustic SFCO sensor (microphone) can be effectively used in medical and biological studies. Note, that reducing the sensor size and modernization of the signal processing interface will enlarge the range of research and diagnostic use of such detectors and may reveal new interesting details of the functioning of various organs and systems of humans and animals. RESEARCH SUPPORT: Precision Sensors/Instrumentation D&P Ltd (www.psi.am, Armenia). HISTOLOGICAL PECULARITIES OF HUMAN PLACENTA AFTER CHRONIC PSYCHO-EMOTIONAL STRESS. Aznauryan AV, Khudaverdyan AD, Heratci Yerevan State Medical University, Yerevan, Armenia INTRODUCTION: The problem of stress influence on fetus, duration of pregnancy and labor has been widely investigated recently. Special place in these investigations can be given to examination of processes, which occur in mother-fetus system and, particularly, in placenta. The aim of this research has been histological examination of placenta of women who were exposed to chronic psychoemotional stress during pregnancy. METHODS: The research involved 10 women with normal pregnancy course and 60 women who were exposed to psychoemotional stress. The state of psychoemotional stress of pregnant women was evaluated based upon anamnesis and testing method of Spilberger, with estimation of anxiety degree. Fragments of a placental tissue were fixed in 10% formalin. Paraffin

11

sections were stained by hematoxylin/eosin, picrotuxin by Van-Gieson. RESULTS: Distinct changes of placental villi vessels and surrounding connective tissue were detected in the placentas of majority of women, who were exposed to psychoemotional stress. It was expressed by the presence of necrosis loci, infarction, fibrinous accretion, luminal occlusion of villi vessels, erythrocytes aggregation and syncytiotrophoblast nuclei accumulations. Described changes represent morphologic substrate of chronic placental insufficiency. Such insufficiency is based upon activation of immunological alteration of placental tissue, as well as implicit changes of vessels, which provide blood flow in mother-placentafetus system. Certain observations show, that fibroblasts and large quantities of collagen fibers were detected in sclerotic villi, which increase pressure upon capillaries. In this case significant extension and induration of placental barrier was observed. Full obliteration of arterioles was observed in separate anchoring villi. In such areas epithelium of villi was partially or fully absent and stroma with compressed capillaries was observed in fibrinoid islets. It is important to mention that the morphological changes were low-grade or even absent in the placentas of some women, who were exposed to psychoemotional stress, which indicate the dependence of discovered changes from individual adaptive abilities of pregnant women. CONCLUSION: Summarizing stated it is possible to conclude, that damages of placental barrier in the mother-fetus system take place in overwhelming majority of women, who were exposed to psycho-emotional stress. POSTER PRESENTATION: DEPRESSION AMONG THE ROMANIAN ADULTS. Abram Z, University of Medicine and Pharmacy, Department of Hygiene, Tirgu-Mures, Romania INTRODUCTION: In the last time it was hardly increased the prevalence of psychical diseases and disorders which reduce work capacity, life quality and life expectancy. The aim of our research is to study the psychical health state of the Romanian adults living in the middle part of the country and the role of some economical, psychological and social factors, especially in relationship with depression. METHOD: The study is based on a complex anonymous questionnaire, including Beck depressive scale, which was completed by a representative sample among adult population. The applied method was a combination between stratification and more-steps sampling. RESULTS: After our results depression is the most common psychical illness with 9.1% diagnosis, but the tendency to depression, the existence of depressive symptoms is much higher than the treated illness. The percentage of suicide attempt among the studied population was 2.9%. We analysed how gender, age, professional and social status, living and working conditions and different social factors are influencing the health state. According to Beck score, it was established a significant difference in the favour of female, elderly people, lower educational level, urban population. CONCLUSION: In our study it is underlined the importance of health promotion and education. It is concluded that improving living standards, modifying in a proper way the lifestyle of the population, we can positively influence the physical and mental health state of the Romanian adult population. PLENARY PRESENTATION 3: FEASIBILITY STUDY OF FEAR CONDITIONING PROTOCOL (Laboras/Sonotrack presentation) Bachdasarian L, Metris B.V., Netherlands SESSION II: PRE-CLINICAL ASPECTS OF PTSD, STRESS AND BEHAVIOR Chairs: Aghajanov MI, Bulthuis R CHANGES IN HIPPOCAMPUS AND CEREBELLUM DEPENDENT BEHAVIOR OF RATS IN HYPOKINETIC CONDITIONS. Navasardyan GA, Yeritsyan NB, Safaryan KS, Grigoryan AS, Yerevan State Medical University, Yerevan, Armenia INTRODUCTION: Sedentary lifestyle became feature of the modern human society, causing a set of various shifts in the body. Besides being a universal risk factor for a host of diseases it is also considered as an important stressor. We consider it as a factor significantly contributing to the allostatic load of a person with corresponding behavioral shifts. The occurring changes in various behaviors significantly depend on the model of movement restriction and are time dependent. The aim of this study was to evaluate the influence of experimental hypokinesia on behavior of rats. METHODS: In a series of studies white male non-linear rats were randomly assigned into 2 main groups: hypokinetic and control groups. Experimental hypokinesia was caused by placing rats in special narrow individual cages (22 h per day) for various terms (2, 7, 15, 30, 45 and 70 days). All rat groups received similar food and water ad libitum. Behavioral tests included X-maze, Y-maze, radial arm maze, active avoidance, bridge- and beam walking, rotorod running, holeboard, walking initiation. RESULTS AND DISCUSSION: At the early stages of hypokinesia (2-15 days - hypokinetic stress) the most pronounced changes were considered to be the increased level of anxiety, decreased locomotion and motor learning. Motor coordination and hippocampal dependent memory were minimally affected. At later stages (45-70 days hypokinetic syndrome) some allostatic balance was reached, general locomotor activity and motor coordination was improved (almost reaching control levels), but hippocampus dependent memory deficits became more prominent (Y- and radial arm mazes and active avoidance). The mentioned time dependent behavioral shifts are driven by various pathophysiological processes,

12

including activation of stress systems and influence of stress hormones; oxidative stress and damage to neurons; glial activation/dysfunction; neuroinflammation and low grade systemic inflammation; impaired afferentation of corresponding neural control centers. The most vulnerable structures out of studied ones were cerebellar Purkinje cells and hippocampal C3 cells. RESEARCH SUPPORT: Yerevan State Medical University, State Scientific Committee. ACTH AND CORTISOL LEVELS IN BLOOD OF PREGNANT RATS AND THEIR OFFSPRING IN THE ACTION OF CHRONIC STRESS. Khudaverdyan DN, Saroyan MY, Khudaverdyan AD, Ghambaryan HK, Heratci Yerevan State Medical University, Yerevan, Armenia INTRODUCTION: The chronic stressor action during prenatal development of the fetus can be one of the reasons for the increasing incidence of the population in the early and late periods of postnatal development. It was found, that prenatal stress causes disturbances in the various systems of the body and, in particular, in neurohormonal regulating mechanisms in mothers and fetal organisms. Based upon this a comparative study of the functional state at the hypothalamic-pituitary-adrenal level of the stress response was done in intact and exposed to chronic emotional stress pregnant rats and their offspring. METHODS: As a stressor we used combined effect of the sound power of 70 dB at 4 kHz, and the light frequency 13Gts, exposure time 350 s, intervals between exposures of 1500 s (6 exposures per day) during pregnancy. The used model of stress was as much as possible close to the stressful conditions of the effects observed in routine life and differs from commonly used in the experiment rigid stress models (e.g., trauma, immobilization), which are rare in the real life. RESULTS AND DISCUSSION: ELISA analysis of ACTH and cortisol levels in the blood showed that stress action within the first 4-5 days of pregnancy was accompanied by a significant increase in the level of hormones in the blood of pregnant rats, and decrease at the end of pregnancy (21- 22 days) in 63% of pregnant rats and a slight increase in 37%, respectively. The content of ACTH and cortisol in the blood of newborn rats (taken 3-5 days after birth) from these groups of pregnant mother rats showed a relatively constant level of hormones in rats born to the first group, with a slight increase in their content of the rats of the second group, respectively. These findings point to the involvement of the mother-fetus system in the implementation mechanisms of stress reaction in pregnant females undergoing stress. The observed tendency of ACTH and cortisol level to decrease in the blood of rats at end of pregnancy can be a consequence of depletion of the corticoid link of stress response in these animals. This, along with the established in some pregnant rats increased levels of hormones in the blood, is indicating the individual characteristics of functioning of the mother-fetus system, and thus the mechanisms for maintaining hormonal balance in the blood of mother and fetus in pregnant females undergoing stress. CONCLUSION: The combined, periodically acting throughout pregnancy sound and light stimuli of moderate strength, although accompanied by typical dynamic stress fluctuations of ACTH and cortisol levels in the blood of pregnant rats, in most cases, does not lead to a significant disruption of the balance in the blood of rats given birth and newborn rats, with the exception of a small part of the examined animals. EXPERIMENTAL ANIMAL MODELS OF PTSD: UPDATE AND FUTURE CHALLENGES. Kalueff AV, Stewart AM, ZENEREI Institute, Slidell, USA The well-recognized fight/freeze/flight response aims to protect an organism from harm associated with danger, such as a terrifying ordeal, threat, abuse, or physiocal harm (NIMH, 2013). However, this normal stress response is markedly affected in post-traumatic stress disorder (PTSD), where exaggerated stress persists long time after the actual danger was present. PTSD is currently affecting approximately 5-10% of general population, rapidly emerging as a key mental health and societal problem. The domain structure of PTSD is based on a strong involvement (and pathogenetic linkage) of both affective and cognitive domains. Animal (experimental) models have become indispensable tools to probe the neurobiology of various brain disorders, including PTSD and its cellular, molecular, and behavioral mechanisms. Several groups of animal models, developed to mimic the symptoms of PTSD, include Pavlovian fear conditioning paradigms, severe physical stress (e.g., restraint), predator exposure-based tests (e.g., cat/cat odor, rat/rat odor exposure, snake exposure, etc). with various degrees of contact, and early developmental stress exposure-based paradigms. To various extend, in addition to robust behavioral phenotypes, these models also model dysregulated fear response neurophysiology, neuromorphological phenotypes (e.g., altered hippocampal plasticity) and endocrine responses associated with PTSD-like states in humans. Collectively, these mounting data provide important practical grounds and conceptual framework for fostering further translational research of PTSD, bridging animal and human findings to increase our understanding of the disorders pathobiology. POSTER PRESENTATION: BAROREFLEX SYSTEM AS A TARGET OF INTERVENTION OF THE ORIGINAL VEGETOTROPIC DRUGS AND THE OBJECT OF MATHEMATICAL MODELING. Shirinyan ME, Shirinyan EA, Mnjoyan Institute of Fine Organic Chemistry (IFOC) STCOPC NAS RA, Yerevan, Armenia

13

INTRODUCTION: Arterial baroreceptor reflex (ABR) is the one of the short-term regulation mechanisms to reduce arterial blood pressure (AP) variability in different extreme situations of the body. ABR system works as a negative feedback mechanism and performs a buffer function by regulation of levels of AP and sympathetic activity. The hyperactivity of the sympathetic nervous system (SNS) in stress situations triggers the increase in AP and develops the concomitant metabolic disorders. In this respect, the discovery of new drugs decreasing or blocking SNS activity effects is topical, and mathematical modeling of the equilibrium state in the ABR system and forecasting of its feedback conditions are very promising. METHODS: The sensitivity of arterial baroreceptors was tested on anaesthetized rats weighing 220-250 g by invasive AP registration and heart rate and R-R interval electrocardiographic registration. The drugs were administered through the catheter implanted into the femoral vein. The test of the sensitivity of arterial baroreceptors and its cardiochronotropic component evaluation were carried out under artificial increase (phenylephrine, i.v. 10-30 mcg/kg) or decrease (sodium nitroprusside, i.v. 515 mcg/kg) in AP. Quantitative evaluation of ABR was conducted using the methods of correlation and regression analysis. To suppress sympathetic activity both known (propranolol, idazoxan) and original adreno- and ganglia blockers, synthesized in the Mndjoyan IFOC were used. Some studies have been conducted on rats previously subjected to sympathectomy by 6-hydroxidophamin. The mathematical model of the equilibrium state regulation of the ABR system has been developed on the basis of hybrid automata. RESULTS AND DISCUSSION: The detected changes in hemodynamic and ABR parameters in rats after the application of aforementioned blockers are discussed in terms of the possibility of ABR regulation when exposed to various parts of the autonomic nervous system. The influence of the inclination angle of the functional curves of the interrelationship between arterial pressure and cardiovascular sympathetic activity on model behavior and time of return into the state of equilibrium has been described. POSTER PRESENTATION: ENHANCING CAPABILITIES FOR STUDY MEMORY AND LEARNING IN THE SHUTTLE BOX AVOIDANCE TASK. Berezhnoy DS, Inozemtsev AN, Department of Physiology of Higher Nervous Activity, Lomonosov Moscow State University, Moscow, Russia The shuttle box active avoidance test has been extensively used in fundamental neurobiological research, regarding fear and learning, and either in experimental psychopharmacology while evaluating new drugs, affecting learning and memory under the stress conditions. Nevertheless, traditional parameters used avoidance reaction rate and reaching the criterion 9 out of 10 correct reactions characterize only the overall learning speed, not allowing the detailed study of the underlying processes. We have developed the methodical approach, allowing us, apart from evaluation of the overall learning effectiveness, to distinguish particular stages of the continuous learning process and characterize corresponding psychophysiological processes. Three distinct stages were identified: the fixation of the memory trace was determined by the following criterion the manifestation of the avoidance reaction in the first trial of the session (generally, is reached in the 2-3 session); the beginning of the habit formation reproduction of the avoidance reaction 3 times in a row (reached by 4-5 session); and the formation of the stable habit reaching the criterion of 75% avoidance reproduction. The use of the suggested learning criterions on the example of antioxidant drugs study revealed that changes, modulating the learning process, could be expressed locally on the stage of memory trace fixation, or stable habit formation, with other stages staying unaffected. The objective of the following analysis was to distinguish behavioral processes, specific for each learning stage. At the first stage, limited by fixation of the memory trace, the correct reaction determination was observed: animal showed the consecutive development of the two reactions escape from the electric shock and, subsequently, avoidance reaction. This process was characterized by the number of trials before the first avoidance manifestation, and dynamics of the reaction latency. After the beginning of habit formation animal behavior expressed the following regularity: the probability of serial avoidance reproduction gradually increased - from 3 to 9-12 reactions in a row. This process reflected the rise in stability of habit error-free reproduction, and continued even after the learning curve (avoidance reaction rate) has reached the plateau. Thus, the developed criteria enable the objective differentiation of the whole learning curve and characterization of the underlying psychophysiological processes, that could be of interest either on their own or in the context of psychopharmacology. POSTER AND DEMONSTRATION (LABORAS, SONOTRACK, DSI) THE NEED FOR MULTI-MODAL AND MULTI-FUNCTIONAL MEASUREMENT SYSTEMS IN LABORATORY ANIMAL RESEARCH AND MATRIX ANALYSIS I. Bachdasarian L, Bulthuis R, Molenwijk E, Karkishenko V, Revyakin A, Metris B.V., Hoofddorp, Netherlands, Scientific Centre Biomedical Technology Moscow, Russia Current trends in the Pharmaceutical industry are requiring not only shorter lead times but also better quality of pre-clinical test results. To achieve this animal experiments will have to collect data from different domains at the same time, for example animal behavior, animal physiology and animal vocalizations. To make this practically possibly automation and integration of different measurement

14

technologies becomes is becoming crucial in preclinical research. Behavior = function {internal stimuli / external stimuli}; Behavior = function {dynamic internal stimulus /from drug effects}; if external factors = constant. Constant environment, modular vivarium and laboratory individually ventilated racks and cages for animals, GLP standard, constant environmental factors (temperature, humidity, ventilation/airflow, odor, no vibration). LABPRODUCTS - provides laboratory products (cages, individual ventilated racks, work benches, change stations, washing machine and many other laboratory (vivarium) Accessories). A constant environment is essential to build reliable behavioral study and analysis. To enhance the quality of the study and have better statistical probability, it is important to analyze many parameters from the same behavior (i.e. group of parameters or matrix). MATRIX METHOD FOR BEHAVIOR ANALYSIS: Proper analysis and statistics of data is very important in invivo experiments. The use of many independent parameters in the automated recognition of a behavior of the animal is therefore crucial. To recognize a behavior automatically, LABORAS applies the Matrix Method involving the analysis of several parameters that are derived from the measurement system. Your specific behavior = matrix / X1, X2, X3 - - -- - Xn; Y1, Y 2, Y 3 - - - - - Y n; E1, E2, E3 - - - - - En/ The above matrix shows an example of the different parameters for a specific behavior. Where X1, X2, X3 - - - - - - Xn, ; E1, E2, E3 - - - - - - En are functions from the specific behavior (e.g. amplitude, frequency, total energy, locomotion energy, locomotion energy / oscillation energy, etc.). Measuring matrix parameters and all kinetic energy in in-vivo experiments is very important to obtain a full ethogram of all behaviors that are shown by the laboratory animal. Traditional methods based on observation or video analysis offer only limited information. The Metris Laboras system enables measurement of all types of kinetic/movement energy while other non-invasive automated systems for behavior detection can only measure the locomotion component of the kinetic energy (e.g. locomotion energy mv/2). In addition, the matrix method and technology used in Laboras provides a way to measure more behaviors and to recognize them completely automatic and more. By combining parameters from different systems the matrix will get better and further improve the quality of the research results. LABORAS - system for fully automatic recognition, recording and analysis of the behavior of small laboratory rodents (rats, mice), based on the analysis of vibration and energy. SONOTRACK-system for recording, playback and visualization of ultrasounds vocalizations in laboratory animals. DSI - system for measuring physiological parameters remotely (without wire measuring pressure, temperature, ECG, EEG, EMG, identification, activity, respiration). SLEEPSIGN / Kissei-SLEEPSIGN software for automatic detection and deep analysis of sleep stages in the animals on the basis of EEG, EMG signals. POSTER PRESENTATION: STRESS-FREE MONITORING OF DEVELOPMENT USING THE LITTER [DAM+PUPS] AS MEASURING UNIT. A PERINATAL STUDY IN RATS WITH METHYLMERCURY. de Groot D, Slieker R, Bulthuis R, Bachdasarian L,TNO Zeist, Zeist, Metris BV, Hoofddorp, Leiden University Medical Center, Leiden, Netherlands Maternal care plays a crucial role in the development of an individual, especially in the response to stress in early and late life. In rats, different types of maternal behavior, particularly grooming and licking, regulate in the offspring the development of emotional, cognitive and endocrine responses to stress. As such, rat models have been widely used to mimic situations in humans and have so contributed to our understanding of the impact of stress. Stress early in life a priori occurs when separating mother and child and was proven to increase the vulnerability of an individual to the toxic potential of environmental chemicals (e.g. drugs and food contaminants). Guidelines for (mandatory) developmental toxicity studies prescribe the daily testing of hundreds of rat pups during the pre-weaning period (i.e. lactation period of the dam) in order to study normal development of senses and reflexes. These tests are considered non-invasive; however, they do greatly disturb the housing and nursing of the litter [dam+pups] in the home cage and induce stress in the litter resulting from separation of pups and dam, and the individual handling of the pups by the bio-technician. Here we present an approach to study the development of rat pups avoiding the stress that is normally induced when separating rat pups from their mother and littermates. In a regulatory toxicity study [US EPA OPPTS 870.6300/8600] with methylmercury (MeHg) known to affect the developing nervous system we explored the usefulness of so called Position Distributions (PDs) of the rat litter in the Laboras system (Metris B.V., Netherlands). The Laboras system is actually developed to measure specific behaviours of an individual animal. In our test set-up, we consider the litter [dam+pups] to be the individual measuring unit. Position Distributions (PDs) were recorded per 24hours from postnatal day (PND) 0 to 21. The PDs represent the activity distribution of the dam and her pups over the home cage. In addition to PDs, we explored with Sonotrack (Metris B.V., Netherlands) the ultrasonic vocalizations emitted by rat pups for communication as marker for development (PND 4 to 18). Body weight and physical /sensory landmarks, conventional markers for development proposed in current test guidelines, were measured for comparison (PND 0 to 21). The results illustrated that the position distributions (DPs) in litters of the control group showed local (bright spot) activity during the first days of the lactation period, and a more generalized (homogenous) activity during the second half of the lactation period. In the MeHg treated group, a delay in the development of the PDs was observed, suggesting delayed development.

15

Ultrasonic vocalizations (USVs) in pups of the control group showed a typical curve of development from PND 4 to 18, peaking on PND 12. In MeHg treated animals a reduced maximum number of calls was observed and a delayed day on which the maximum number of calls was reached. Over time, a dose-related shift of the normal pattern of calling was clearly visible. This was also found in the minimum frequency of calls, showing a delayed day of minimum of frequency of calls. Sensory landmarks in the MeHg groups showed tendency towards developmental delay whereas physical landmarks appeared earlier (not significant); all landmarks were reached at significantly lower body weight. These results support the idea that position distributions (PDs) and ultrasonic vocalisations (USVs) may form fast, animal-friendly markers to study normal development and developmental (neuro)toxicity. Combination of Laboras and Sonotrack (Metris B.V., Netherlands) may form a basic test set-up for a stress-free automated recording of normal development and effects thereon by external stimuli, using the rat litter [dam and pups] as the principle measuring unit. RESEARCH SUPPORT: Dutch Ministries of Health, Welfare and Sports, the Social Affairs and Employment, and the Economic Affairs.

DAY 2, SATURDAY, 21 SEPTEMBER 2013

SESSION III: MOLECULAR-GENETIC ASPECTS OF COGNITION, PHYSIOLOGY AND STRESS Chairs: Kalueff AV, Sargisyan S, Strekalova T COMPARATIVE CHARACTERISTICS OF DIFFERENT SUBPOPULATIONS OF THE IMMUNE COMPLEXES IN PSYCHO-NEUROLOGICAL DISORDERS: IMPLICATION TO DISEASE-RELATED MOLECULAR PATHOMECHANISMS. Chavushyan A, Hovhannisyan L, Mkrtchyan G, Hovsepyan M, Arakelyan A, Hakobyan S, Khoyetsyan A, Petrosyan M, Harutyunyan K, Musaelian R, Ayvazyan V, Hovhannisyan G, Aroutiounian R, Boyajyan A, Institute of Molecular Biology NAS RA, Armenian Society of Biological Psychiatry affiliated to the World Federation of Societies of Biological Psychiatry, "rtsakh" Scientific Center of Nagorno-Karabakh, Yerevan State University, Yerevan, Armenia INTRODUCTION: Numerous studies implicate upregulated inflammation and autoimmune sensitization in pathogenesis of many psycho-neurological disorders. Excess formation of the immune complexes (IC) and their different subpopulations in circulation are indicators of these processes. OBJECTIVES: The present report provides overview of our recent studies related to identification of the presence of abnormal subpopulations of IC in the blood of patients with posttraumatic stress disorder (PTSD), schizophrenia (Sch), and ischemic stroke (IS), determination of their levels, and comparative analysis of their structural and functional characteristics. SUBJECTS AND METHODS: Patients with PTSD, Sch, IS, and healthy volunteers (HV) were enrolled in these studies approved by the Ethical Committee of the Institute of Molecular Biology NAS RA (IRB #00004079). A broad range of preparative and analytical procedures and in vitro tests was applied including different electrophoretic and blotting procedures, cell culture technology, enzyme-linked immunosorbent assay, cytotoxicity and genotoxicity assays, etc. ANOVA statistical analysis was used to evaluate differences between groups. P values less than 0.5 were considered as significant. RESULTS: Total levels of IC in the blood serum of PTSD-, Sch-, and IS-affected subjects were significantly higher than in HV group (p<0.05). In all three groups of patients the presence of pathogenic small-size IC, absent in HV, were found. In patients with PTSD and Sch the increased levels of opsonin-containing IC were found. In addition, in patients with Sch and IS the presence of cryoglobulins was detected. Further isolation, purification of the abnormal subpopulations of the IC from the blood of study subjects and investigation of their immunochemical composition, ability to induce cytokine expression by peripheral blood mononuclear cells, as well as cytotoxic and genotoxic properties revealed significant differences in these parameter depending on pathology. CONCLUSIONS: PTSD, Sch and IS are associated with the excess production of IC and their different subpopulations in circulation. Each selected diseased condition has its specific spectrum of abnormal subpopulations of IC. Immunochemical composition of the abnormal subpopulations of IC reflects disease-related changes in homoeostasis of the human body. DISC1 REGULATION OF D-SERINE PRODUCTION IS SPECIFIC TO ASTROCYTES. Abazyan S, Ma TM, Abazyan B, Nomura J, Yang C, Seshadri S, Sawa A, Snyder SH, Pletnikov MV, Department of Psychiatry and Behavioral Sciences, The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA, Division of Molecular Medical Science, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan INTRODUCTION: Disrupted in schizophrenia 1 is a protein that is encoded by the DISC1 gene in humans. Porteus and associates discovered that disruption of two genes (DISC1 and DISC2) by a balanced chromosomal translocation (1;11) in a Scottish family co-segregates with schizophrenia, major depression and bipolar disorder. Our prior study demonstrates that two psychiatric genetic risk factors, Disrupted-in-schizophrenia (DISC1) and serine racemase (SR), partner to regulate production of D-

16

serine (Ma et al, 2013). Although our findings suggest that this interaction may take place in astrocytes, a direct functional evaluation of DISC1-SR interaction in neurons was incomplete. METHODS: We overexpressed dominant-negative mutant DISC1 or knocked down endogenous DISC1 using RNAi lentiviral vector and measured expression of SR and D-serine production in primary neurons or astrocytes or hippocampal brain tissue samples of mice that express mutant DISC1 selectively in astrocytes or neurons. RESULTS AND DISCUSSION: Our data confirm the hypothesis that DISC1 regulates SR expression and associated D-serine production predominantly in astrocytes. The interaction between these two psychiatric genetic risk factors in astrocytes but not neurons further supports the critical role of astrocytes in the pathophysiology of NMDA synaptic transmission in schizophrenia and related mental disorders. RESEARCH SUPPORT: R01MH-083728 EFFECT OF ELECTRIC FOOT SHOCK STRESS ON TRYPTOPHAN CONCENTRATION IN PLASMA AND RED BLOOD CELLS OF DIFFERENT AGE RATS. Nevoia A, Ciochina V, Garaeva S, Postolati G, Redcozubova G, Institute of Physiology and Sanocreatology MAS, Chisinau, Moldova INTRODUCTION: There are convincing data that long-term stress with disturbance of adaptation leads to stress disorders with cognitive and psychic declines. Tryptophan is the precursor for the neurotransmitter 5-hydroxytryptamine (5-HT) which have a major involvement in stress regulation. It is present in bound and free form in the blood, where the concentration is controlled by albumin binding to tryptophan. A large proportion of free tryptophan is storage in red blood cells (RBC) which are involved in depository and inter-organ transport of amino acids. The aim of this study was to explore the effect of chronic stress on tryptophan concentration in plasma and RBC in rats of different age. METHODS: 3week, 12- and 24- month old rats were stimulated by electric foot shock for 6 weeks. The shocked mice received 0.6 mA electric shocks in a communication box 2 times/min. during 30 min twice a day. The blood plasma and RBC concentrations of free tryptophan and the branched chain amino acids (BCAA), valine, leucine, izoleucine, which compete with tryptophan for entry into the brain across the blood brain barrier, have been investigated by the method of liquid chromatography. RESULTS AND DISCUSSION: The plasma concentration of free tryptophan was found to increase in all the groups (1.6 and 1.9 fold in adult and young animals, respectively) and more significantly in old ones (2.8 fold; p < 0.05). Simultaneously were noticed a considerable increase in tryptophan content in the RBC, especially in young and old rats (3.8- and 4.5-fold, respectively; p < 0.05) that might indicate an intensification inter-organ transport of tryptophan. The augmentation of free tryptophan concentration observed in young rats plasma, together with the rise in the plasma concentration rate of free tryptophan/BCAA suggest an increase in the rate of transport of tryptophan across the blood-brain barrier and consequently to an increase in the rate of synthesis of 5-hydroxytryptamine (5-HT) in the brain. CONCLUSION: Obtained dates denote the age-specific peculiarities of changes in tryptophan content under prolonged stress and point out the high responsiveness of young rats. MOLECULAR AND GENETIC INDICATORS OF APOPTOTIC AND SYNAPTIC PLASTICITY ALTERATIONS IN POSTTRAUMATIC STRESS DISORDER AND SCHIZOPHRENIA. Boyajyan A, Avetyan D, Atshemyan S, Sukiasyan S, Gevorgyan A, Mkrtchyan G, Zakharyan R, Chavushyan A, Melkumova M, Torosyan S, Institute of Molecular Biology NAS RA; Armenian Society of Biological Psychiatry WFSBP, Russian-Armenian (Slavonic) University, Stress Center MLSA RA, Psychiatric Medical Center MH RA, Yerevan, Armenia INTRODUCTION: Posttraumatic stress disorder (PTSD) and schizophrenia (Sch) are severe polygenic psychiatric diseases characterized by cognitive impartment, which may result from apoptotic and synaptic plasticity dysfunction. OBJECTIVES: In the present study blood levels of annexin-a5 and complexin-2 proteins involved in apoptosis and synaptic plasticity were determined and functional single nucleotide polymorphisms (SNPs) of genes encoding these proteins (ANXA5 and CPLX-2) were evaluated in patients with PTSD (DSM-IV-TR code: 309.81) and paranoid form of Sch (DSM-IV-TR code: 295.30) in comparison to healthy subjects (HS). SUBJECTS AND METHODS: In total, 100 patients with PTSD (Karabakh combat veterans), 260 Sch-affected subjects, and 260 HS were involved in this study. The study was approved by the Ethical Committee of the Institute of Molecular Biology NAS RA (IRB 00004079). The experiments were performed using blood serum/plasma and genomic DNA samples of study subjects. Methodological design was mainly based on application of the enzymelinked immunosorbent assay and polymerase chain reaction with sequence-specific primers. Data were evaluated using Hardy-Weinberg equilibrium, Pearsons Chi-square test, Bonferroni multiple correction approach, Mann-Whitney U-test, Kruskal-Wallis H-test, Dunn's multiple comparison test and Spearman correlation analysis. RESULTS: The obtained results indicated that: (1) PTSD is characterized by hypoactivity of apoptosis manifested by decreased blood evels of annexin-a5; (2) PTSD is characterized by decreased synaptic plasticity manifested by decreased plasma levels of complexin-2; (3) alterations in apoptosis rate and synaptic plasticity in PTSD are interrelated; (4) PTSD is associated with the SNP rs1366116 of CPLX-2; (5) the rs1366116*T mutant allele of CPLX-2 represents risk factor for PTSD; (6) decreased blood levels of complexin-2 protein in PTSD result from a prevalence of the rs11575945*T mutant allele of CPLX-2 in PTSD-affected subjects; (7) Sch is characterized by

17

hyperactivation of apoptosis manifested by increased blood levels of annexin-a5; (8) Sch is associated with the SNP rs11575945 of ANXA5 as well as the SNPs rs1366116 and rs3892909 of CPLX-2; (9) the rs11575945*T mutant allele of ANXA5 and rs1366116*T mutant allele of CPLX-2 represent risk factors for Sch, whereas rs3892909*T mutant allele of CPLX-2 decreases risk for development of Sch; (10) increased blood levels of annexin-a5 in Sch result from both hyperactivation of apoptosis and a prevalence of the rs11575945*T mutant allele of ANXA5 in Sch-affected subjects; (11) the age of the first manifestation of Sch in subjects homozygous for rs11575945*T mutant allele of ANXA5 is lower than in carriers of two other genotypes of the rs11575945 polymorphism of this gene. CONCLUSIONS: Alterations in apoptotic rate and synaptic plasticity are involved in pathogenesis of PTSD and Sch. Annexin-a5 and complexin-2 proteins blood levels may be considered as molecular markers of altered apoptosis and synaptic plasticity in PTSD and Sch. CPLX-2 may be nominated as a candidate gene for both PTSD and Sch, and ANX5 may be nominated as a candidate gene for Sch. ROUND TABLE: PTSD (POST-TRAUMATIC STRESS DISORDER) AND GENETICS: HOW GENOCIDE AND WAR INFLUENCE GENETICS AND BEHAVIOR Chairs: Bachdasarian L, Tadevosyan AS, Margaryan S PTSD AND SIGNALS EFECTS ON HUMANS AND ANIMALS. Bachdasarian L, Metris BV, Hoofddorp, Netherlands INTRODUCTION: The Signal influence from environment on the human and animal body is very high. The influence of the signal can be positive or negative. For example, negative stimuli are genocide, war, terrorism; and positive stimuli include music, dancing or sport competitions. The human body receives stimulus energy from the environment, and starts to analyze and make choices. Sometimes the choices are right, other times they are wrong. In some cases we see responses in form of behaviors or psychic effects. Environment affects the animal body just like it has effects on humans. For example different ultrasonic vocalizations create different types of stimuli on animals leading to different choices and responses. On a global scale, we need to work towards decreasing the negative stimulus and maximizing the positive stimulus. GOALS: Analyze the PTSD influences and effects. Make special scale of PTSD disease and effects, dependency from the traumatic events (TE) and traumatic brain injury (TBI) (signal stimulus from environment and effectors factors). METHODS: I researched history and several study areas of PTSD (from 1915 Armenian genocide, World War II, Veterans study from Vietnam, Rwanda, Iraq). I further investigated many PTSD influences and effects. The official 17 symptoms of PTSD can be placed into 3 broad groups. Re-experiencing: intrusive memories, nightmares, flashbacks, triggered distress; Avoidance: isolation, withdrawal, emotional numbing, detachment, memory gaps; and Hyperarousal: insomnia, irritability, anger outbursts, poor concentration, hypervigilance, exaggerated startle. Beyond the official diagnosis are associated features that result in poor occupational and social function. They include depression, suicidal ideation, alcohol and drug abuse, guilt, shame, inability to trust, over-controlling, few or no close relationships, extreme isolation, unemployment, divorce, domestic violence, and child abuse. If a person experiences long term PTSD than there is a higher chance that a person may develop Chronic PTSD with Hormonal and Genetic influences. The National Vietnam Veterans Readjustment Study (NVVRS) was conducted by the U.S. government to better understand the PTSD and psychological effect of being in the Vietnam War. The findings from this study were alarming. At the time of the study (middle to late 1980s), among Vietnam veterans, approximately 15% of men and 9% of women were found to currently have PTSD. Approximately 30% of men and 27% of women had PTSD at some point in their life following Vietnam. These findings, obtained approximately a decade after the end of the Vietnam War, found that for many veterans, their PTSD had become a chronic (that is, persistent and long-lasting) condition. In fact, fourteen years after this study, many of these veterans were interviewed again, and it was found that a substantial proportion continued to have PTSD, as well as a number of other difficulties, such as physical health problems, marital problems, and psychological difficulties. European researchers have identified a gene that is linked to improved memory, but also to increased risk of post-traumatic stress disorder (PTSD). RESULTS: Table 1. Average percentage of PTSD from war
After (TE), year 0-1 1-3 3-5 5-10 Total PTSD % 6-8% 25-38% 45-57% 43-55% Men% 5 20 35 30 Women% 3 18 22 25 long term effect% not visible 20 33 30

10-20 40-45% 25 20 28 Percentage of men is higher than women; The long term effect and the chance of chronic PTSD is growing

18

Table 2. Statistic age and PTSD percentage from war

Age, years 0-5 5-12 12-18 18-27 27-45 Total PTSD % 2 4 6 37 28 men 1 1 2 25 15 women 1 3 4 12 13 8

45-70 23 15 Average age of 18-27; the risk of PTSD is higher than older age Table 3. Average percentage of PTSD from Genocides After (TE), years 0-1 1-3 3-5 5-10 Total PTSD % 33-48% 68-78% 65-77% 50-65% Men% 25 40 35 30 Women% 23 28 30 25

Long term effect% not visible 30 45 35

10-20 45-55% 23 20 29 All percentages are very high; The chance of chronic PTSD and Genetic effect is very higher than from war Table 4. Statistic age and PTSD percentage from genocide Age, years 0-5 5-12 12-18 18-27 27-45 Total PTSD % 5 14 16 31 19 men 2 6 8 14 9 women 3 8 8 17 10

45-70 15 8 7 Average age of 18-27y. the risk of PTSD is higher than older, but in this case the percentage of PTSD growing from child

Figure: PTSD Evolution THE LONG-TERM IMPACT OF PTSD: From the study PTSD count found that almost 3 decades 1200 after the Vietnam War, many 1000 1960 veterans continued to 1950 800 2010 1920 1930 1990 600 experience problems with 1940 2000 1970 1980 400 PTSD. At the initial interview, 200 1900 1910 0 approximately 12% had 1880 1900 1920 1940 1960 1980 2000 2020 PTSD. Fourteen years later, ye ar the rates of PTSD had dropped only slightly to approximately 11%. Those who had experienced high levels of combat exposure were most likely to have PTSD at both interviews. Veterans who continued to have PTSD 14 years after their first interview were found to have considerably more psychological and social problems. They reported lower satisfaction with their marriage, sex life, and life in general. They also indicated having more parenting difficulties, higher divorce rates, lower happiness, and more physical health complaints, such as fatigue, aches, and colds. After the long-term PTSD, there are increased chance of Chronic PTSD and also the chance of genetic effects. GENETIC INFLUENCE OF PTSD: SCIENTISTS IDENTIFY GENETIC CHANGES THAT MAY INCREASE RISK OF PTSD: Natural disaster, violent crime and war are traumatic experiences for anyone to live through, yet some people recover quickly from these events while others struggle with the flashbacks and hypervigilance that characterize posttraumatic stress disorder (PTSD). Why? Scientists say they have pinpointed certain genetic changes that may make some people more vulnerable to PTSD than others. The finding could one day make it possible to screen people for increased risk of developing the disorder. Working with several generations of 12 families who survived the devastating Spitak earthquake in Armenia in 1988, Dr. Armen Goenjian and his colleagues at the UCLA, found that variations in genes that code for the brain chemical serotonin can affect how profoundly people are affected by trauma. Serotonin is implicated in a number of moodPTSD variation
PTSD count

19

related disorders, including depression and anxiety. The neurotransmitter helps regulate sleep, mood and feelings of satisfaction and satiety, and doctors often prescribe medications that increase the amount of serotonin available in the brain to treat both depression and anxiety. Because PTSD often occurs together with depression, Goenjian wondered whether serotonin-regulating genes might also be involved in PTSD. To find out, he took samples from 200 adults who survived the Armenian earthquake and conducted genomic analyses on three specific genes that regulate serotonin production. In his previous work on potential genetic contributors to PTSD, Goenjian reported that about 40% of vulnerability to the mental disorder was inherited. In the current study, he isolated two possible variations of genes that code for an enzyme involved in serotonin production that could contribute to this generational propensity to PTSD. Together, the gene changes may account for 7% to 8% of the variance in severity of PTSD symptoms. Goenjian is under no illusion that hes found the primary genetic driver behind PTSD; its likely that the disorder is the result of a combination of multiple genetic variants. But, he says, were pleased to have it. The next step is to pursue this finding in a larger population and probably other ethnic and racial groups, and to pursue other genes also. Still, the serotonin-related genes do provide a promising avenue for developing a way to screen people for increased risk of developing PTSD, and possibly to protect them from situations where they might experience severe emotional and physical trauma. If someone has a vulnerability to developing PTSD, then we might be careful in assigning them to a task, such as going to a high combat zone where he might be exposing himself to a situation where he could be at risk of developing [the disorder], says Goenjian. The findings could also help researchers create PTSD treatments that address changes in serotonin and help people who are more vulnerable to traumatic experiences recover from them more quickly. The Division has several projects that address the relationship between PTSD and aging, a research priority of the National Center. Division staff are collaborating with investigators at VA Boston Healthcare system to study the effects of exposure to traumatic events and other stressors on mental and physical health among aging military Veterans. Another, newly funded, project aims to organize and make available existing longitudinal data sets that can be used to address research questions about the impact of trauma on the course of aging. A multinational, multidisciplinary group of experts will advise this effort and launch the research agenda. SCALE OF PTSD: Scale 1 Genocide; Scale 2 - violence for child; Scale 3 war. The Scale 1 and 2 has higher chance to get chronic PTSD and Genetic effects. REFERENCES: Departments of Medicine (Seal), Epidemiology and Biostatistics (Sen), and Psychiatry (Marmar), University
of California, San Francisco, and Health Services Research and Development Research Enhancement Award Program (Bertenthal), San Francisco VA Medical Center, San Francisco (Seal, Miner, Sen, and Marmar). Dr. A Goenjian and his colleagues at the University of California, Los Angeles/Read more: www.healthland.time.com/2012/04/03/scientists-identifygenetic-changes-that-may-increase-risk-of-ptsd/#ixzz2WNAtiX00; www.nji.nl; www.nih.gov; www.nimh.nih.gov; www.tno.nl; www.cbs.nl; www.medicalnewstoday.com; www.ptss-support.nl; www.ggzingeest.nl; www.ptss.nl; www.genetics.nl; www.dare.uva.nl/document/336690; www.uva.nl/nieuws-agenda/nieuws/uva-nieuws; www.metris.nl/en/products/laboras

PRODUCT PRESENTATION (METRIS AND DSI) THE NEED FOR MULTI-MODAL AND MULTI-FUNCTIONAL MEASUREMENT SYSTEMS IN LABORATORY ANIMAL RESEARCH AND MATRIX ANALYSIS II. Bachdasarian L, Bulthuis R, Molenwijk E, Gaburro S, Meuth P, Metris B.V., Netherlands, Data Sciences International, St. Paul, USA, Department of Neurology, Inflammatory Disorders of the Nervous System and Neurooncology, Institute of Physiology I, Westflische Wilhelms-Universitt Mnster, Germany Current trends in the Pharmaceutical industry requires new translational approaches for pre-clinical test. Those aspects can be achieved by animal experiments in which not only a single variable (e.g. behavior) at the time is analyzed but rather a multidimensional approach (physiology+behavior) is applied. Therefore, automation and integration of different measuring technologies become the crucial aspects in this process. Behavior = function {internal stimuli / external stimuli}; Behavior = function {dynamic internal stimulus /from drug effects}; if external factors = constant. THE ANIMAL MODEL FOR PTSD RESEARCH IS FEAR CONDITIONING PROTOCOL: In this protocol, mouse models are very important. There are two ways of measuring fear responses in mice: Startle response and Freezing behavior. Metris BV developed two special algorithms for automated Startle and Freezing detection (See Video: presentation). Having an automated detection system is not sufficient at the behavioral level for excluding false results (e.g. sleeping phases -> less movement confounded as freezing). Therefore, the behavioral response needs to be integrated and synchronized with physiological parameter (e.g. EEG,ECG, BP, Datasciences Int., The best way to do so would be using Laboras system (for behavioral study), DSI (for Physiological parameters) and Sonotrack (for ultrasounds vocalization study). See more details on Metris B.V.s web-site www.metris.nl. STANDARDIZING THE ANALYSIS OF CONDITIONED FEAR IN RODENTS: A MULTIDIMENSIONAL SOFTWARE APPROACH: Data comparability between different laboratories strongly depends on the individually applied analysis method. This factor is often a critical source of variation in rodent phenotyping and has never been systematically investigated in Pavlovian fear conditioning paradigms. In such tests, emotional learning/memory performances or efficacy of novel anxiolytic compounds are assessed. In rodents fear is typically quantified in terms of freezing duration via manual observation or automated systems. While manual analysis includes biases like tiredness, anticipation or inter-personal scoring

20

variability, computer-assisted systems were yet unable to distinguish between freezing and immobility. Consequently, the novel software called MOVE follows a semi-automatized approach that pre-filters video sequences of interest for the final human judgment. Furthermore, MOVE allows integrating additional data sources from (force based LABORAS system platform (Metris B.V., Netherlands; www.metris.nl), ECG) to reach the most accurate and precise results. MOVE supports single- or multiangle video recordings with cost effective webcams, standard laboratory equipment or a hybrid solution. The integrated manual key logger and internal video player complement this all-in-one software solution. Calculating the inter-laboratory variability of manual freezing evaluation revealed significantly different freezing scores in 2 out of 6 laboratories. This difference was minimized when all experiments were analyzed with MOVE. Multi-angle video analysis compared to the single camera approach reached up to 15% higher accuracy and two-fold higher precision. Integrating a LABORAS system platform data source further improved the previous results. Based on that8 CD-1 mice were implanted telemetry devices (Data Sciences International; www.datasci.com) for HR recording and underwent Pavlovian fear conditioning. By using the data integration capability of Laboras system the combined signal of all data sources allowed distinguishing freezing-related events from those of pure quiescence. SESSION IV: NEUROBIOLOGY OF STRESS IMPACT ON BRAIN PLASTICITY AND MEMORY Chair: Yeritsyan NB THE IMPACT OF EARLY PHARMACOLOGICAL TRAUMA ON BRAIN ANATOMY IN GERBILS. Lehmann K, Institute for General Zoology and Animal Physiology, Friedrich-Schiller University, Jena, Germany Traumatic experiences in early childhood are considered as causative factors in the etiology of psychiatric diseases. Their consequences are, however, not compelling: Later living conditions may favor or prevent the outbreak of a manifest disease. We studied this interaction of early trauma and rearing environment in an animal model in the Mongolian gerbil. As a traumatic event, we used the injection of a single high dose of methamphetamine on postnatal day (PD) 14. As an environmental factor, animals were born and reared either in an impoverished environment with isolated housing after weaning, or socially in an enriched, large arena. In young adulthood (PD90), neuroanatomical assessments were done in cortical and subcortical areas. Especially, monoaminergic and cholinergic fiber densities were assessed, and prefrontocortical efferents were studied by anterograde tracing. We could show that both methamphetamine intoxication and impoverished rearing as single factors lead to widespread, but mostly balanced changes in fiber densities and cortical connections. If both challenges convened, however, balances were disrupted, and the observed changes were often opposite to those after single challenges. Our results show that the consequences of early traumatic experiences can to a certain degree be absorbed by a favorable rearing environment. An irksome environment during adolescence, in contrast, can result in severely disturbed brain maturation. The observation that this is reflected in a mis-wiring of prefrontal cortical and subcortical efferents suggests that our model shares neuroanatomical features with human schizophrenia. SPECIAL LECTURE III: STRESS, EARLY LIFE EVENTS AND MEMORY FORMATION. Krugers H, University of Amsterdam, Swammerdam Institute for Life Sciences, Amsterdam, Netherlands Memories for emotionally arousing and fearful events are generally well retained. From the evolutionary point of view this is a highly adaptive behavioral response aimed to remember relevant information. However, fearful memories can also be inappropriately and vividly (re)expressed, such as in posttraumatic stress disorder (PTSD). The memory formation of emotionally arousing events is largely modulated by corticosteroid hormones and neurotransmitters which are released during and after exposure to stressful events. In this presentation I will address that 1) Corticosteroid hormones dynamically regulate surface expression of AMPA receptors and synaptic function which is crucial for synaptic plasticity and memory formation. Behavioral studies in mutant mice suggest that corticosteroid hormones promote emotional memory formation via regulation of AMPA receptors; 2) Corticosteroid hormones and beta-adrenergic receptors interact to regulate AMPA receptor function; 3) Early postnatal life is an important period which determines synaptic function and memory later in life: negative experiences during early life lastingly regulate dendritic complexity, synaptic plasticity and emotional memory formation. ENVIRONMENTAL FACTORS TO INDUCE OR TO RELEAVE STRESS AND DEPRESSIVE TRAITS: A STUDY ON MICE. Anthony DC, Chernopyatko AS, Strekalova T, Department of Pharmacology, University of Oxford, Oxford, UK, Timantti AB, Stockholm, Sweden, Department of Neuroscience, School for Mental Health and Neuroscience, Maastricht University, Maastricht, Netherlands Dealing successfully with environmental challenges involves the initiation of multiple molecular, cellular and behavioral responses. Individuals who are unable to cope or are chronically stressed often develop psychiatric disorders, such as depression and anxiety. On another hand, some environmental factors can have protective effects. Given the vast numbers of people suffering from these disorders, it is of

21

major importance that the mehcanisms underlying the impact of stress on the brain are investigated. A chronic stress paradigm comprising exposure to predation, tail suspension and restraint induces a depressive syndrome in C57BL/6J mice that occurs in some, but not all, animals. Compared to resilient animals, susceptible mice display a unique molecular changes, including upregulated expression of tumour necrosis factor alpha (TNF), the 5-HT transporter (SERT), indoleamine-2,3-dioxygenase (IDO) in a number of brain regions. At the cellular level, increased numbers of Iba-1-positive microglial cells were also present in the prefrontal area of susceptible animals compared to resilient animals. Most of these changes can be reversed by various antidepressant therapies, and, interestingly, some environmental factors. Consequently, this altered profile provides a rationale for exploring antiinflammatory, and possibly, TNF-targeted therapy for major depression. This can help to develop better treatment strategies for depression and anxiety and hopefully in a not too distant future to prevent development of these disorders. LEARNING-ASSOCIATED STRESS: PLASTICITY UNDERLYING MEMORY. Yeritsyan NB, Department of Neurophysiology, Leibniz Institute for Neurobiology, Magdeburg, Germany Unveiling the influence of learning-associated stress on the synaptic plasticity in behaving animals appears to be an efficient approach in understanding the mechanisms of memory formation and its stress-induced impairment. Recent research has shown that functional synaptic plasticity events such as long-term potentiation (LTP) and depression (LTD) underlie the memory formation in experimental behavioral tasks mostly involving a stressful experience. This approach, when developed further, may support the understanding of a complex effect of stress administrated prior to or associated with encoding process on later remembering of a given event in humans. With this regard, in my talk I will address the mechanisms of hippocampal plasticity induced by swim stress in water maze taskuntrained rats. The role of the ventral tegmental area (VTA), a dopaminergic structure involved in the behavioral reinforcement and emotional processing, will be highlighted as in the modulation of swim stress-induced CA1-LTD, as in the mechanisms of spatial memory consolidation. The interference between hippocampal long-term memories in highly emotional one-trial inhibitory avoidance task and complex spatial water-maze task with hidden platform will be discussed in a context of 'ruthless competition' between synaptic events and brain circuits, which leads to the formation of a strong memory trace for emotionally arousing experience along with an impairment of other memories. Such interaction is believed to contribute to the pathogenesis of intrusive flashbulb memories and retrograde amnesia in patients with posttraumatic stress disorder. ARMENIAN MUSEUM OF GENOCIDE PRESENTATIONS IMPLICATIONS OF GENOCIDE DENIAL ON THE VICTIM GROUP: THE ARMENIAN CASE. Gzoyan E, International Relations, LLM, Yerevan, Armenia WHO ARE THEY? ARMENIAN WOMENS EXPERIENCE DURING AND AFTER THE GENOCIDE. Aleksanyan A, Armenian Genocide Museum-Institute, Yerevan, Armenia

PUBLISHED COMMUNICATION: IMPACT OF ACOUSTIC STRESS ON NEURONAL ACTIVITY OF THE CENTRAL AND LATERAL AMYGDALOID AND DORSAL RAPHE NUCLEI IN RATS. Arajyan G, Sargsyan R, Yeritsyan N, Department of Physiology, Abovyan Armenian State Pedagogical University, Yerevan, Armenia, Orbeli Institute of Physiology, National Academy of Sciences of Armenia, Yerevan, Armenia, Department of Neurophysiology, Leibniz Institute for Neurobiology, Magdeburg, Germany nalysis of a spontaneous firing activity (SFA) of neurons in the central (CAM) and lateral (LAM) nuclei of amygdala as well as the dorsal raphe (DR) nuclei has been performed under normal conditions and after a 2.5-h lasting acoustic stress. It has been shown that after acoustic stress the activity of all brain structures except the DR was challenged with the expressed alterations in the LAM. In intact animals the SFA parameters in the CAM and DR were quite similar, while in the LAM several differences (e.g. smaller average firing frequency, high coefficient of variation, etc.) were detected. Furthermore, we have investigated the changes in the CAM and LAM SFA electrolytic lesions of the dorsal raphe nucleus. Destruction of either the DR resulted in two-fold decrease in average firing frequency of CAM neurons, whereas in the LAM the two-fold increase of the same parameter was registered. The overall changes in the SFA were more profound for the lateral amygdaloid nucleus. The statistical analysis has revealed the positive correlation in the direction of SFA changes in the CAM and LAM after the acoustic impact and the DR lesion of, which assume a possible involvement of the DR in the brain response to the acoustic stress.

22

Metris BV, Hoofddorp, Netherlands, el: +31235623400, Fax:+31235623425, e-mail: info@metris.nl, web: www.metris.nl
Metris is a leading manufacturer of advanced system for animal behavior analysis (in-vivo experiments) that are sold globally. Main products are: LABORAS, SONOTRACK and Ultrasounds SmartChamber. LABORAS is an advanced system that automates behavior scoring, registration and analysis of small laboratory animals. The system tracks positions and simultaneously identifies more than 18 validated stereotypical and normal behaviors in mice and rats. Laboras does not use video or infrared beams! There are over 200 publications about the use of Laboras by several leading researchers, pharmaceutical companies, CROs and leading universities from around the world. SONOTRACK is an advanced system to record, analyze and playback ultrasound vocalizations. The system is highly valued for research in Anxiety, Stress, Memory, Learning, Pain, Sexual related, Safety Pharmacology, Developmental (Neuro) Toxicity and Social Interaction tests. Sonotrack is the best ultrasound vocalization system on the market today because of its full spectrum USV recording (15 kHz to 125 kHz) characteristics, extremely low noise, long duration recording capability and reliable detection of rodent calls fully automatically! Ultrasounds SmartChamber provides a sound isolated, ventilated and light controlled environment to perform high performance ultrasonic vocalization experiments. The chamber includes an ultrasonic microphone and the interior of the chamber effectively removes sound echoes, external noise, sounds and magnetic fields. SmartChamber can be seamlessly integrated with our product Sonotrack. In the CIS countries, Metris sells modular vivaria and laboratory cabins and several other vivarium and laboratory equipment, including cages, Individual Ventilated Cages (IVC racks), workstations, washing machines, wireless equipment for animal identification and temperature registration, systems for wireless measurement of physiology parameters (ECG, EEG, EMG, Blood pressure, Temperature, respiratory, activity ) and sleep analysis software. In addition several other systems for molecular and cellular analysis are being offered. Metris is distributor for DataSciences International (DSI-telemetry), LabProducts, Bio Medical Data systems (BMDS) and Kissei Comtec. - M . Bachdasarian L, Bulthuis R, Molenwijk E, Karkishenko V, Revyakin A, Metris B.V., Netherlands, Scientific Centre Bio Medical Technology, Moscow, Russia , . , , , . (). = { / }; = { / }; / = . - , GLP- , (, , , , ). LABPRODUCTS (, , , , , () ); . , ( ). : . . , LABORAS , , . = / X1, X2, X3 - - - - ; Y1, Y 2, Y 3 - - - - - Y , E1, E2, E3 - - - - En / . X1, X2, X3 - - - - - - - Xn, ; E1, E2, E3 - - - - - - - En (, , , , / , ..). invivo (. ) , . ,

23

 , . LABORAS Metris , (mv/2) . , , Laboras, , . , . LABORAS - , (, ), . SONOTRACK - , . DSI ( , , ECG, EEG, G, , , ). SLEEPSIGN/Kissei- SLEEPSIGN EEG, G .

OOO
(in-vivo ), . : LABORAS, SONOTRACK, . LABORAS , (, ), . 18 . Laboras ! 200 Laboras , , CRO . SONOTRACK - , , . , , /, , , , () , . Sonotrack - USV (15-125 ), , ! (Ultrasounds SmartChamber) , . , , . - Sonotrack. , , , IVC , , , , (ECG, EEG, EMG, Blood pressure, Temperature, respiratory, activity) . , . Metris DataSciences International (DSI-telemetry), LabProducts, Bio Medical Data systems (BMDS) Kissei Comtec.

24

THE INTERNATIONAL STRESS AND BEHAVIOR SOCIETY (ISBS)

Established in 2007 President: Allan V. Kalueff, PhD (2013-2015) Vice-President: Viktor M. Klimenko, MD, PhD (2013-2015) ISBS Executive Office Coordinator: Nataliya A. Zinevych www.stress-and-behavior.com info@stressandbehavior.com ISBS is the international society of experts working with a wide range of topics in the field of translational neuroscience, neurobehavioral sciences, biopsychology and biopsychiatry, with a particular focus on stress, stress-related neurobehavioral phenotypes, their neural, molecular and genetic mechanisms, as well as stress-evoked neuropsychiatric disorders. Anyone with an interest in stress-related human or animal behaviors, neurobehavioral disorders and their mechanisms, wishing to join the International Stress and Behavior Society can do so by paying dues of USD $100.00 regular member or $60.00 student member for a three-year term. Payment can be made following sending the e-mail form and payment request to the ISBS Secretariat at info@stressandbehavior.com. Once the form and the payment have been received, you will receive a membership confirmation. Membership: Regular membership dues are $ 100.00 for the period of three years, or $ 60.00 for the period of one year. Student (undergraduate and graduate) membership dues are $60.00 for the period of three years. Membership period starts January 1 of each year. New members registered from May 2013 will automatically be covered for the period of 2013-2016. Regular membership benefits include a $50.00 discount for registration for any of the ISBS Conferences, symposia, workshops and summer schools. Student members will benefit from a $25 discount for registration for any of the ISBS Conferences, symposia, workshops and summer schools. ISBS Membership application form (please fill in and send by e-mail to the ISBS Secretariat at info@stressandbehavior.com, with the subject ISBS Membership request ) Name, Family name: Position/Title: Institute/Company: Category: please select one: Regular member, 3-year term ($ 100.00) Regular member, 1 year term ($ 60.00) Student member, 3-year term ($ 60.00) Address (affiliation): City: Postal code: Country: Phone, Fax: E-mail address: www:

25

Please join our future conferences:

4th International Regional Neuroscience and Biological Psychiatry Conference "STRESS AND BEHAVIOR" (North America)
June 22-24, 2014, New Orleans, LA, USA

21st International Neuroscience and Biological Psychiatry Conference "STRESS AND BEHAVIOR"
May 16-19, 2014, St. Petersburg, Russia

5th International Regional Neuroscience and Biological Psychiatry Conference "STRESS AND BEHAVIOR" (North America)
June 22-24, 2015, Miami, FL, USA

www.stress-and-behavior.com
26