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Development in Immunoprophylaxis against Rabies for Animals and Humans 2 2 Rabies is a fatal neurological disease and a persistent global problem. It is spread primarily by domestic dogs but other canid, viverrid (skunks and raccoons) and chiropteran species are considered as the most efficient vectors of the disease. Since dogs are the main perpetuator of rabies, special attention has to be given to bring all the dogs including unauthorized stray dogs under immunization umbrella in order to control rabies. Vaccination is the only way to combat the disease before and after exposure or infection as there is no treatment available once the symptoms have appeared. After the first crude nerve tissue vaccine developed by Pasteur in 1885, a number of rabies vaccines for animal and human use have been developed with varying degree of safety and efficacy over the years. Presently, cell culture based inactivated rabies vaccines are largely used in most of the parts of the world. However, these vaccines are too expensive and unaffordable for vaccination of people and animals in developing countries. The comparatively cheaper inactivated nerve tissues vaccines can cause serious side-effects such as autoimmune encephalomyelitis in inoculated animals and production has been discontinued in several countries. Although attenuated live vaccines can efficiently elicit a protective immune response with a smaller amount of virus, they sometimes can cause rabies in the inoculated animals by its residual virulence or pathogenic mutation during viral propagation in the body. New-generation rabies vaccines generated by gene manipulation although in experimental stage may be a suitable alternative to overcome the disadvantages of the live attenuated vaccines. So, awareness must be created in general public about the disease and the cell culture based vaccines available in the market should be recommended for wide scale use to prevent and control this emerging and reemerging infectious disease in foreseeable future. 3 21 Sukdeb Nandi Virology Laboratory, Center for Animal Disease Research and Diagnosis (CADRAD), Indian Veterinary Research Institute (IVRI) India Manoj Kumar Virology Laboratory, Center for Animal Disease Research and Diagnosis (CADRAD), Indian Veterinary Research Institute (IVRI) India LyssavirusRabies virusRabiesVaccinationZoonoses 29.pdf Knobel DL, Cleaveland S, Coleman PG, Fevre EM, Meltzer MI, Miranda ME, et al. Reevaluating the burden of rabies in Africa and Asia. Bull WHO 2005;83(5):360-368. ##Wilde H, Khawplod P, Khamoltham T, Hemachudha T, Tepsumethanon V, Lumlerdacha B, et al. Rabies control in South and Southeast Asia. Vaccine 2005;23(17-18):2284-2289. ##Hampson K, Dushoff J, Bingham J, Bruckner G, Ali YH, Andy D. 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The Effect of Human Chorionic Gonadotropin Treatment on Recipient Mouse Germ Cell Proliferation Following Spermatogonial Stem Cell Transplantation of Neonatal Donor Mice 2 2 Spermatogonia are the male germ line stem cells whose life long expansion is needed for permanent production of spermatozoa. The present study was designed to examine the effect of hCG treatment on germ cell proliferation following stem cell transplantation in mice. Spermatogonial stem cells were isolated from neonatal mice testes and characterized by alkaline phosphatase, immunoreactivity and morphological analysis. hCG was injected into normal and cell transplanted mice. We then evaluated the testosterone levels and cell number in normal mice. After that, cyclin B1 gene expression was investigated in transplanted mice. Different doses of busulfan were injected to investigate the effects of chemotherapy on morphological criteria and preparation of recipient mice for transplantation. In this report we show proliferative potential of spermatogonial stem cells after cytotoxic treatment, transplantation efficiency by semi-quantitative RT-PCR, and hCG effect on stem cell regeneration in normal mice and following cell transplantation. The results indicate that spermatogonial stem cells can proliferate after transplantation, and the efficiency of their transplantation depends on hormonal treatment. Therefore, hormonal treatment after stem cell transplantation will be a powerful avenue for increasing the efficiency of transplantation and fertility restoration. 23 35 Mohammad Mehdi Akhondi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Reza Akbarzadeh Najar Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mahmood Jeddi-Tehrani Immune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska Institute Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR SwedenIran Mohammad Reza Sadeghi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Amir-Hassan Zarnani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Hodjattallah Rabbani Immune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska Institute Sweden Sheida Salehkhou Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Leila Eini Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Fatemeh Hoseinzadeh Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mahnaz Heidari Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran BusulfanCell proliferationhCGTransplantation 27.pdf Kubota H, Avarbock MR, Brinster RL. 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The effect of three alkylating agents on the seminiferous epithelium of rodents. II. Cytotoxic effect. Virchows Archiv B Cell Pathol 1970;4(4):276-282. ##Towns R, Azhar S, Peegel H, Menon KM. LH/hCG-stimulated androgen production and selective HDL-cholesterol transport are inhibited by a dominant-negative CREB construct in primary cultures of rat theca-interstitial cells. Endocrine 2005;27(3):269-277. ##Troiano L, Fustini MF, Lovato E, Frasoldati A, Malorni W, Capri M, et al. Apoptosis and spermatogenesis: evidence from an in vivo Model of testosterone withdrawal in the adult rat. Biochem Biophys Res Commun 1994; 202(3):1315-1321. ##Dunkel L, Perheentupa J, Apter D. Kinetics of the steroidogenic response to single versus repeated doses of human chorionic gonadotropin in boys in prepuberty and early puberty. Pediatr Res 1985;19(1):1-4. ##Kerr JB, Sharpe RM. Focal disruption of spermatogenesis in the testis of adult rats after a single administration of human chorionic gonadotrophin. 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Production and Characterization of Mouse Monoclonal Antibodies Recognizing Multiple Subclasses of Human IgG 2 2 Different IgG subclass profiles are produced in response to different antigenic stimuli in a variety of diseases. IgG subclass levels may reflect disease severity. Quantification of IgG subclasses depends on the availability of specific Monoclonal antibodies (MAbs). In the present study seven hybridoma clones producing MAbs reactive with multiple subclasses of human IgG were established. Splenocytes from Balb/c mice immunized with Fc fractions of human IgG1 or IgG2 myeloma proteins were fused with mouse myeloma cells. Fused cells were selected and cloned by limiting dilution assay. Antibody secreting cells were screened by Enzyme-linked immunosorbent assay (ELISA) and the specificity of secreted MAbs was further analyzed, using a panel of purified human myeloma paraproteins of different IgG subclasses by ELISA and immunoblotting. Cross-reactivity to immunoglobulins (Igs) of other species was studied by indirect ELISA using serum samples collected from 9 animals. The MAbs were found to react with triple IgG subclasses, including IgG1,2,4 (n=4) and IgG1,2,3 (n=3). Immunoblotting studies revealed recognition of linear (n=4) or conformational (n=3) epitopes by these MAbs. The most abundant cross-reactivity (71.4%) was observed with monkey Ig while no cross-reactivity was detected with hen and cat sera. The MAbs mostly displayed a restricted pattern of cross-reactivity and one of them did not bind to any of the animal sera tested. The affinity constant of 3 MAbs was measured by ELISA. Based on the data obtained from this study, mouse MAbs reactive with multiple human IgG subclasses are directed to a variety of immunogenic epitopes, mostly shared with IgG of other species. These MAbs are valuable tools for purification of non-reactive IgG subclasses through negative affinity chromatography. These MAbs could also provide an opportunity for epitope mapping of the Fc region of IgG, as well as serological phylogenetic studies. 37 45 Fatemeh Hajighasemi Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Medicine, Shahed University Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Medicine, Shahed University IranIran Fazel Shokri Department of Immunology, School of Public Health, Tehran University of Medical Sciences Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Department of Immunology, School of Public Health, Tehran University of Medical Sciences Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR IranIran ELISAHybridomaIgG IsotypeMonoclonal antibodySubclass 28.pdf Hajighasemi F, Khoshnoodi J, Shokri F. Development of two murine monoclonal antibodies recognizing human nG1m(a)-like isoallotypic markers. Hybridoma 2008;27(6):473-479. ##Hussain A, Pankhurst T, Goodall M, Colman R, Jefferis R, Savage CO, et al. Chimeric IgG4 PR3-ANCA induces selective inflammatory responses from neutrophils through engagement of Fcgamma receptors. Immunology 2009;128:236-244. ##Bartel G, Wahrmann M, Exner M, Regele H, Schillinger M, Horl WH, et al. Determinants of the complement-fixing ability of recipient presensitization against HLA antigens. Transplantation 2007;83(6):727-733. ##Rynnel-dagoo B, Freud A, Hammarstrom L, Oxelius V, Persson MA, Smith CI. Pneumococcal antibodies of different immunoglobulin subclasses in normal and IgG subclass deficient individuals of various ages. Acta Otolaryngol 1986;101(1-2):146-151. ##Soderstrom T, Enskog A, Samuelsson BE, Cedergren B. Immunoglobulin subclass (IgG3) restriction of anti-P and anti-Pk antibodies in patients of the rare p blood group. J Immunol 1985;134(1):1-3. ##Saha S, Mazumdar T, Anam K, Ravindran R, Bairagi B, Saha B, et al. Leishmania promastigote membrane antigen-based enzyme-linked immunosorbent assay and immunoblotting for differential diagnosis of Indian post-kala-azar dermal leishmaniasis. J Clin Microbiol 2005;43(3):1269-1277. ##Beenhouwer DO, Yoo EM, Lai CW, Rocha MA, Morrison SL. Human immunoglobulin G2 (IgG2) and IgG4, but not IgG1 or IgG3, protect mice against Cryptococcus neoformans infection. Infect Immun 2007;75(3):1424-1435. ##Kaneko Y, Nimmerjahn F, Ravetch JV. Anti-inflammatory activity of immunoglobulin G resulting from Fc sialylation.Science 2006;313(5787):670-673. ##Abd El-Aal AA, El-Arousy MH, El-Gendy AM, Tunkul Ael-K, Ismail SA, El-Badry AA. Early post-treatment immunoglobulin profile in human schistosomiasis. J Egypt Soc Parasitol 2005;35(1):167-180. ##Tangteerawatana P, Montgomery SM, Perlmann H, Looareesuwan S, Troye-Blomberg M, Khusmith S. Differential regulation of IgG subclasses and IgE antimalarial antibody responses in complicated and uncomplicated Plasmodium falciparum malaria. Parasite Immunol 2007;29:475-483. ##Suresh L, Kumar V. Significance of IgG4 in the diagnosis of mucous membrane pemphigoid. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2007;104(3):359-362. ##Windle HJ, Ang YS, Athie-Morales V, McManus R, Kelleher D. Human peripheral and gastric lymphocyte responses to Helicobacter pylori NapA and AphC differ in infected and uninfected individuals. Gut 2005;54(1):25-32. ##Morais CG, Soares IS, Carvalho LH, Fontes CJ, Krettli AU, Braga EM. IgG isotype to C-terminal 19 kDa of Plasmodium vivax merozoite surface protein 1 among subjects with different levels of exposure to malaria in Brazil. Parasitol Res 2005;95(6):420-426. ##Chevrier MC, Chateauneuf I, Guerin M, Lemieux R. Sensitive detection of human IgG in ELISA using a monoclonal anti-IgG-peroxidase conjugate. Hybrid Hybridomics 2004;23(6):362-367. ##Burton DR, Gregory L, Jefferis R. Aspects of the molecular structure of IgG subclasses. Monogr Allergy 1986;19:7-35. ##Hajighasemi F, Shokri F. Generation and characterization of mouse hybridomas secreting monoclonal antibodies specific for human IgG3. AJMB 2009;1(1):19-26. ##Loirat MJ, Gourbil A, Frioux Y, Muller JY, Blanchard D. A murine monoclonal antibody directed against the Gerbich 3 blood group antigen. Vox Sang 1992;62(1):45-48. ##Hajighasemi F, Gharagozlou S, Ghods R, Khoshnoodi J, Shokri F. Private idiotypes located on light and heavy chains of human myeloma proteins characterized by Monoclonal antibodies. Hybridoma 2006;25(6):329-335. ##Hajighasemi F, Saboor-Yaraghi AA, Shokri F. Measurement of affinity constant of anti-human IgG monoclonal antibodies by an Elisa-based method. Iranian J Immunol 2004;1(3):154-161. ##Beatty JD, Beatty BG, Vlahos WG. Measurement of monoclonal antibody affinity by non-competitive enzyme immunoassay. J Immunol Meth 1987;100(1-2):173-179. ##Hajighasemi F, Gharagozlou S, Ghods R, Roohi A, Khoshnoodi J, Shokri F. Generation and characterization of a mouse monoclonal antibody with specificity similar to staphylococcal protein A (SPA). Hybrid Hybridomics 2003;22(1):33-39. ##Jefferis R, Lowe J, Ling NR, Porter P, Senior S. Immunogenic and antigenic epitopes of immunoglobulins. I. Cross-reactivity of murine monoclonal antibodies to human IgG with the immunoglobulins of certain animal species. Immunology 1982;45(1):71-77. ##Reimer CB, Phillips DJ, Aloisio CH, Moore DD, Galland GG, Wells TW, et al. Evaluation of thirty-one mouse monoclonal antibodies to human IgG epitopes. Hybridoma 1984;3(3):263-275. ##Lowe J, Bird P, Hardie D, Jefferis R, Ling NR. Monoclonal antibodies (McAbs) to determinants on human gamma chains: Properties of antibodies showing subclass restriction or subclass specificity. Immunology 1982;47(2):329-336. ##Girkontaite I, Leckiene M, Mauricas M. Immunochemical study of human immunoglobulin G Fc region. Cancer Biother Radiopharm 1996;11(1):87-96. ##Recht B, Frangione B, Franklin E, Van LE. Structural studies of a human ?3 myeloma protein ( Goe ) that binds Staphylococcal protein A. J Immunol 1981;127(3):917-923. ##Boyle MDP. Bacterial immunoglobulin-binding proteins (Vol 1). San Diego: Academic press; 1990,17-28. ##Eliasson M, Olsson A, Palmcrantz E, Wiberg K, Inganäs M, Guss B, et al. Chimeric IgG-binding receptors engineered from staphylococcal protein A and streptococcal protein G. J Biol Chem 1988;263:4323-4327. ##Burton DR, Gregory L, Jefferis R. Aspects of the molecular structure of IgG subclasses. Monogr Allergy 1986;19:7-35. ##Hamilton RG, Morrison SL. Epitope mapping of human immunoglobulin-specific murine monoclonal antibodies with domain- switched, deleted and point-mutated chimeric antibodies. J Immunol Meth 1993;158(1):107-122. ##

Intermittent Feeding Attenuates Clinical Course of Experimental Autoimmune Encephalomyelitis in C57BL/6 Mice 2 2 Multiple Sclerosis (MS) is an autoimmune inflammatory, demyelinating disease of human central nervous system. Experimental Autoimmune Encephalomyelitis (EAE) is the commonly used animal model of MS. Calorie restriction has been found to reduce inflammation and autoimmune responses and promote neuroprotection. In this study we evaluated the effects of intermittent feeding protocol of the calorie restriction in a mouse model of EAE. Fifty four female mice (C57BL/6) were used in this study. The animals were divided into two dietary groups: ad libitum (AL) (n=29) with free access to food and water and intermittent feeding (IF) (n=25) with access to food on alternate days. After 8 weeks, EAE was induced in animals by immunization with MOG antigen (Hooke labs, Lawrence, MA, USA) subcutaneously. AL and IF groups were then further divided into two groups each: AA (ad libitum until the end of study) (n=16) and AI (subjected to intermittent feeding regimen after immunization day) (n=13). The IF group was divided into II (continued intermittent feeding regimen until the end of study) (n=13) and IA (changed to AL regimen after immunization day) (n=12). All the animals were behaviorally monitored for 35 days after immunization and observed daily for the signs and severity of disease with EAE scoring scale [0-5] and cumulative disease index (CDI) score. Intermittent feeding significantly reduced the incidence of EAE in IF groups (AI 0%, II 18.5%, IA 22.2%, p<0.05). In addition, intermittent feeding significantly delayed the onset of EAE in AI group (p<0.05) and also, intermittent feeding significantly reduced the severity of disease in II and IA groups (AA vs. II, p<0.05 & AA vs. IA p<0.05) groups. The CDI was also significantly reduced in intermittent feeding fed groups [AI, II and IA compared to AA group (P<0.05, <0.01, <0.05 respectively)]. Intermittent feeding regimen protocol of the calorie restriction significantly suppressed EAE incidence, induction, and severity. The results of this study suggest possible role of intermittent feeding in the treatment of Multiple Sclerosis patients. 47 52 Laya Kafami Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences Iran Mohsin Raza Applied Neuroscience Research Center, Baqiyatallah University of Medical Sciences Applied Neuroscience Research Center, Baqiyatallah University of Medical Sciences Iran Alireza Razavi Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences Iran Abbas Mirshafiey Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences Iran Mansooreh Movahedian Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Iran Mohammad Reza Khorramizadeh Department of Medical Biotechnology, School of Advanced Medical Technologies, Tehran University of Medical Sciences Department of Medical Biotechnology, School of Advanced Medical Technologies, Tehran University of Medical Sciences Iran Experimental autoimmune encephalomyelitisIntermittent feedingMiceMultiple sclerosis 30.pdf Sospedra M, Martin R. Immunology of multiple sclerosis. Annu Rev Immunol 2005;23:683-747. ##Lutton JD, Winston R, Rodman TC. Multiple sclerosis: etiological mechanisms and future directions. Exp Biol Med 2004;229(1):12-20. ##Pugh TD, Klopp RG, Weindruch R. Controlling caloric consumption: protocols for rodents and rhesus monkeys. Neurobiol Aging 1999;20:157-165. ##Kealy RD, Lawler DF, Ballam JM, Mantz SL, Biery DN, Greeley EH, et al. Effects of diet restriction on life span and age-related changes in dogs. J Am Vet Med Assoc 2002;220(9):1315-1320. ##Jolly CA. Dietary restriction and immune function. J Nutr 2004;134:1853-1856. ##Iwashige K, Kouda K, Kouda M, Horiuchi K, Takahashi M, Nagano A, et al. Calorie restricted diet and urinary pentosidine in patients with rheumatoid arthritis. J Physiol Anthropol Appl Human Sci 2004;23(1):19-24. ##Jolly CA. Diet manipulation and prevention of aging, cancer and autoimmune disease. Curr Opin Clin Nutr Metab Care 2005;8(4):382-387. ##Muthukumar A, Zaman Kh, Lawrence R, Barnes JL, Fernandes G. Food restriction and fish oil suppress atherogenic risk factors in lupus-prone (NZB x NZW) F1 mice. J Clin Immunol 2003;23 (1):23-33. ##Muthukumar AR, Jolly CA, Zaman Kh, Fernandes G. Calorie restriction decreases proinflammatory cytokines and polymeric Ig receptor expression in the submandibular glands of autoimmune prone (NZB x NZW)F1 mice. J Clin Immunol 2000;20 (5):354-361. ##Sun D, Krishnan A, Su J, Lawrence R, Zaman Kh, Fernandes G. Regulation of immune function by calorie restriction and cyclophosphamide treatment in lupus-prone NZB/NZW F1 mice. Cell Immunol 2004;228(1):54-65. ##Esquifino AI, Cano P, Jiménez-Ortega V, Cutrera RA, Cardinali DP. Experimental allergic encephalomyelitis in male Lewis rats subjected to calorie restriction. J Physiol Biochem 2004; 60(4):245-252. ##Esquifino AI, Cano P, Jiménez-Ortega V, Fernández Mateos M, Cardinali DP. Immune response after experimental allergic encephalomyelitis in rats subjected to calorie restriction. J Neuroinflammation 2007;4(1):6. ##Piccio L, Stark JL, Cross AH. Chronic calorie restriction attenuates experimental autoimmune encephalomyelitis. J Leukoc Biol 2008;84(4):940-948. ##Lee J, Seroogy KB, Mattson MP. Dietary restriction enhances neurotrophin expression and neurogenesis in the hippocampus of adult mice. J Neurochem 2002;80(3):539-547. ##Mattson M. Neuroprotective signaling and the aging brain: take away my food and let me run. Brain Res 2000;886(1-2):47-53. ##Bruce-Keller AJ, Umberger G, Robert McFall R, Mattson MP. Food restriction reduces brain damage and improves behavioral outcome following excitotoxic and metabolic insults. Ann Neurol 1999; 45(1): 8-15. ##Duan W, Mattson MP. Dietary restriction and 2-deoxyglucose administration improve behavioral outcome and reduce degeneration of dopaminergic neurons in models of Parkinson's disease. J Neurosci Res 1999;57(2):195-206. ##Yu ZF, Mattson MP. Dietary restriction and 2-deoxyglucose administration reduce focal ischemic brain damage and improve behavioral outcome: evidence for a preconditioning mechanism. J Neurosci Res 1999;57(6):830-839. ##Mattson MP, Duan W, Guo Z. Meal size and frequency affect neuronal plasticity and vulnerability to disease: cellular and molecular mechanisms. J Neurochem 2003;84(3):417-431. ##Policies on the use of animals and humans in neuroscience research. Guidelines and policies [cited; Available from: http://www.sfn.org/index.aspx?pagename=guidelinesPolicies_Use of Animals and Humans. ##21. Becher B, Durell BG, Noelle RJ. Experimental autoimmune encephalitis and inflammation in the absence of interleukin-12. J Clin Invest 2002:110(4):493-497. ##Garay L, Gonzalez Deniselle MC, Lima A, Roig P, De Nicola AF. Effects of progesterone in the spinal cord of a mouse model of multiple sclerosis. J Steroid Biochem Mol Biol 2007;107(3-5):228-237. ##Chacon F, Esquifino AI, Perell M, Cardinali DP, Spinedi E, Alvarez MP. 24-hour changes in ACTH, corticosterone, growth hormone, and leptin levels in young male rats subjected to calorie restriction. Chronobiol Int 2005;22(2):253-265. ##

Editorial 2 2 New advances in Genetic Technologies are changing the lives of millions of people around the world in important ways. These advances have also raised difficult ethical and legal questions for policy makers in many countries. Therefore, there is a great interest and need by experts in both governmental and non-governmental institutions to have a deeper understanding of the issues involved for establishing the necessary societal rules to regulate the use of genetic technologies in the fields of Medicine, Veterinary Science and Agriculture. To address these issues and provide a forum for a scientific discussion at a national level, the Avicenna Research Institute is planning to hold a conference in November 2010, entitled 'Genetics: Law, Ethics and Psychology'. The conference will particularly focus on the use of new genetic technologies and its impacts on the society from the legal and ethical point of view. In view of the fact that better understanding of the genetic basis of human behavior and physiology is imperative to comprehend the more complex topics of the conference, the genetics of human behavior will also be discussed in the convention. The conference will aim to address the specific areas of concern in the use of genetic technologies in human health as follows: • Assisted Reproduction Techniques (e.g. in-vitro fertilization)- ART are used to help fertility problems. Ethical issues are around the creation, selection, and disposal of embryos. These technologies can also require the use of sperm, eggs, or wombs from other women who are unrelated to the real parents and are not expected to play a role in raising the child. • Pre-implantation Genetic Diagnosis – In this technique an embryo at 6-10 cell stage can be tested and selected for or against a specific sex, disease and physical condition. Although, this method can be used for treatment purposes, however the ethical issues are around the use of this technology by parents to select a specific baby based on personal desires. • Cloning and Stem cells - Cloning is an essential tool of modern biology which has led to important drugs and new therapies. Cloning also has helped the understanding of genetic basis of human development and disease. Cloning has a potential to be used in producing a lifetime supply of therapeutic stem cells that are genetically matched to a patient. The ethical issues around cloning concern the production and destruction of a two-to-four-day-old embryo to make a line of embryonic stem cells. Another, concern is assuring that women donating eggs for research give proper informed consent. Some fear that a cloned embryo could be implanted into a woman resulting in a baby; a cloned human being. • Animal and Plant Cloning – Cloning animals and plants for specific purposes are becoming possible with new genetic technologies. Specific recombinant proteins to be used as therapeutic drugs are being produced in some animals and specific plants with certain characteristics are now possible to be made and some are currently available in the market for human use. The ethical issues around the animal and plant cloning are the consequence of releasing such new species in the environment and its impact on the society and human health. • Biobank – Biospecimens are being stored in public and private repositories and contain genetic material to identify gene variations associated with human diseases and lead to diagnostic tests and targeted treatments for specific diseases. Ethical issues are around the methods used in obtaining informed consent, protect privacy and disclose of research results, ownership of biospecimens as intellectual property and the ethical use of the biospecimens. • New frontiers in genetics research–There have been many new genetic technologies developed and there are plans to be applied in human health. • The research areas such as Human Behavior, Epigenetics, Nutrigenomics are some of the topics that are planned to be addressed in the forum. I look forward to your active participation in the conference and receive articles examining the issues related to ethical and legal aspects of Medical Biotechnology. 52 52 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 157.pdf ####

Flow Cytometric Analysis of 4-HPR-induced Apoptosis and Cell Cycle Arrest in Acute Myelocytic Leukemia Cell Line (NB-4) 2 2 In many acute leukemias, normal differentiation does not occur. However, in many cell lines derived from hematologic malignancies, differentiation or programmed cell death (apoptosis) can be induced by variety of agents including: Vitamin analogs, demethylating agents, cyclic AMP analogs and anti-proliferative agents. To the best of our knowledge there has been not any study specifically to analyze apoptotic and anti-proliferative effects of 4-HPR (a vitamin analog) in NB-4 cell line. To test whether this drug has activity in acute myeloid leukemia (AML), we first analyzed the anti-proliferative effect of 4-HPR in one AML cell line (NB-4) using MTT Assay. Next we tested whether this drug induced apoptotic cell death. The ability of this compound to induce apoptosis of cancer cells was examined by Annexin V-FITC Assay using Flow cytometry. We also analyzed the cell cycle progression by PI staining using flow cytometry. Using MTT assay, NB-4 cells exhibited increased inhibition of proliferation at micromolar concentrations of 4-HPR at 24, 48 and 72 hrs post treatment. Flow cytometry analysis indicates that 4-HPR is a potent inducer of in vitro apoptotic cell death, and cell cycle analysis revealed an increase in S phase population. In total, the results indicate that 4-HPR is a strong inhibitor of AML cell proliferation and a potent inducer of in vitro apoptotic cell death. Further studies are required to evaluate the in vitro effects of 4-HPR in AML blasts derived from AML patients. 53 61 Shahrzad Soleymani Fard Biology Department, Faculty of Basic Science, Science and Research Branch, Islamic Azad University Biology Department, Faculty of Basic Science, Science and Research Branch, Islamic Azad University Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mohammad Mehdi Akhondi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mehrdad Hashemi Molecular Genetic Department, Tehran Medical Branch, Islamic Azad University Molecular Genetic Department, Tehran Medical Branch, Islamic Azad University Iran Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Iran Acute myelocytic leukemiaApoptosisCell differentiationFlow cytometry4-HPR 26.pdf McKenzie SB. Advances in understanding the biology and genetics of acute myelocytic leukemia. Clin Lab Sci 2005;18(1):28-37. ##Stone RM, O'Donnell MR, Sekeres MA. Acute myeloid leukemia. Hematology 2004;98-117. ##Lovino CS, Camacho LH. A classification and treatment update. Clin J Oncol Nurs 2003;7(5):535-540. ##Gale RP. Progress in acute myelogenous leukemia. Ann Intern Med l984;101:702-705. ##Weinstein HJ, Mayer RJ, Rosenthal DS, Coral FS, Camitta BM, Gelber RD. Chemotherapy for acute myelogenous leukemia in children and adults: VAPA update. Blood 1983;62(2):315-319. ##Foon KA, Gale RP. Therapy of acute myelogenous leukemia. Blood Reviews 1992;6(1):15-25. ##Deangelo DJ. The treatment of adolescents and young adults with acute lymphoblastic leukemia. Hematology 2005;123-130. ##Darwiche N, Hatoum A, Dbaibo G, Kadara H, Nasr R, Abou-Lteif G, et al. N-(4-hydroxyphenyl) retinamide induces growth arrest and apoptosis in HTLV-I-transformed cells. Leukemia 2004;18:607-615. ##Sun SY, Yue P, Lotan R. Induction of apoptosis by N-(4-hydroxyphenyl) retinamide and its associ-ation with reactive oxygen species, nuclear retinoic acid receptors, and apoptosis related genes in human prostate carcinoma cells. Mol Pharmacol 1999;55(3):403-410. ##Ulukaya E, Pirianov G, Kurt MA, Wood EJ, Mehmet H. Fenretinide induces cytochrome c release, caspase 9 activation and apoptosis in the absence of mitochondrial membrane depolar-isation. Cell Death Differ 2003;10:856-859. ##Yazdanparast R, Moosavi M, Mahdavi M, Lotfi A. Guanosine 5'-triphosphate induces differentiation dependent apoptosis in human leukemia U937 and KG1 cells. Acta Pharmacologica Sinica 2006;27(9):1175-1184. ##Sun SY, Hail N Jr, Lotan R. Apoptosis as a novel target for cancer chemoprevention. J Natl Cancer Inst 2004;96(9):662-672. ##Supino R, Crosti M, Clerici M, Warlters A, Cleris L, Zunino F, et al. Induction of apoptosis by fenretinide (4HPR) in human ovarian carcinomacells and its association with retinoic acid receptor expression. Int J Cancer 1996;65(4):491-497. ##Hail Jr N, Kim H J, Lotan R. Mechanisms of fenretinide-induced apoptosis. Apoptosis 2006 ;11:1677-1694. ##Paulson JD, Oldham JW, Preston RF, Newman D. Lack of genotoxicity of the cancer chemopreventive agent N-(4-hydroxyphenyl) retinamide. Fundam Appl Toxicol 1985;5(1):144-150. ##Lotan R. Retinoids and apoptosis: implications for chemoprevention and therapy. J Natl Cancer Inst 1995;87(22):1655-1657. ##Faderl S, Lotan R, Kantarjian HM, Harris D, Van Q, Estrov Z. N-(4 Hydroxylphenyl) retinamide (fenretinide, 4-HPR), a retinoid compound with antileukemic and proapoptotic activity in acute lymphoblastic leukemia (ALL). Leuk Res 2003;27(3):259-266. ##Maurer BJ, Metelitsa LS, Seeger RC, Cabot MC, Reynolds CP. Increased of ceramide and induction of mixed apoptosis/ necrosis by N-(4-hydroxyphenyl) retinamide in neuroblastoma cell lines. J Natl Cancer Inst 1999;91(13):1138-1146. ##Tamaoki T, Nomoto H, Takahashi I, Kato Y, Morimoto M, Tomita F. Staurosporine, a potent inhibitor of phospholipid/ Ca++dependent protein kinase. Biochem Biophys Res Commun 1986;135(2):397-402. ##Kruman I, Guo Q, Mattson MP. Calcium and reactive oxygen species mediate staurosporine-induced mitochondrial dysfunction and apoptosis in PC12 cells. J Neurosci Res 1998;51(3):293-308. ##DiPietrantonio AM, Hsieh T-C, Olson SC, Wu JM. Regulation of G1/ S transition and induction of apoptosis in HL-60 leukemia cells by fenretinide (4-HPR). 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Development of a Latex Agglutination Test as a Simple and Rapid Method for Diagnosis of Trichomonas vaginalis Infection 2 2 Trichomoniasis is a worldwide infection and due to its complications rapid and accurate diagnosis of infection especially in pregnant women is very important. In this study, development of a latex agglutination test using native antigens for rapid diagnosis of trichomoniasis is investigated. Trichomonas vaginalis was harvested from TYIS33 culture medium and anti Trichomonas vaginalis antiserum was raised in rabbits. Salt precipitation method was used for antibody purification. Polyesteren latex particles coated with purified antibody and used for detection of Trichomonas vaginalis. Clinical samples of vaginal discharge were collected from 500 women and examined for Trichomonas vaginalis by using wet mount, culture and latex agglutination tests. Sensitivity and specificity of latex test was determined considering culture as golden standard. Sensitivity and specificity of latex agglutination test was 100% and 81% and those of wet mount were 33.3% and 100%, respectively. Positive and negative predictive values of latex agglutination test were 6% and 100%, respectively. Due to inconvenient sensitivity and specificity of the latex agglutination test developed in this study, further work is recommended to improve the test. 63 66 Hossein Yousofi Darani Department of Parasitology, Cell and Molecular Research Center, Faculty of Medicine, Shahrekord University of Medical Sciences Department of Parasitology, Cell and Molecular Research Center, Faculty of Medicine, Shahrekord University of Medical Sciences Iran Firuzeh Ahmadi Department of Parasitology, Shahrekord University of Medical Sciences Department of Parasitology, Shahrekord University of Medical Sciences Iran Nozhat Zabardast Department of Parasitology, Shahrekord University of Medical Sciences Department of Parasitology, Shahrekord University of Medical Sciences Iran Hossein Ali Yousefi Department of Parasitology, Isfahan University of Medical Sciences Department of Parasitology, Isfahan University of Medical Sciences Iran Hedayat Shirzad Department of Immunology, Faculty of Medicine, Shahrekord University of Medical Sciences Department of Immunology, Faculty of Medicine, Shahrekord University of Medical Sciences Iran Latex agglutination testSensitivitySpecificityTrichomonas vaginalis 31.pdf Alzanbagi NA, Salem HS, Al Braiken F. Trichomoniasis among women with vaginal discharge in Jeddah city, Saudi Arabia. J Egypt Soc Parasitol 2005;35(3):1071-1080. ##Sutton M, Sternberg M, Koumans EH, McQuillan G, Berman S, Markowitz L. The prevalence of Trichomonas vaginalis infection among reproductive age women in the United States 2001-2004. Clin Infect Dis 2007;45(10):1319-1326. ##Holmes King T. Sexually transmitted diseases. 4th ed. New York: McGraw Hill; 2008. ##Rein MF, Chapel TA. Trichomoniasis, candidiasis and the minor venereal diseases. Clin Obstet Gynecol 1975;18(1):73-88. ##Krelger JN. Urological aspects of trichomoniasis. Invest Urol 1981;18(8):411-417. ##Spence MR, Hollander DH, Smith J, McCaig L, Sewell D, Brockman MMT. The clinical and aboratory diagnosis of Trichomonas vaginalis infection. Sex Trans Dis 1980;7(4):168-171. ##Martens GM, Faro S, Soper D. Infectious disease in women. 1st ed. Philadelphia: Saunders; 2001. ##McCann JS. Comparison of direct microscopy and culture in the diagnosis of trichomoniasis. Br J Vener Dis 1974;50:450-452. ##Audo Sarkodie Y, Opoku BK, Danso KA, Weiss HA, Mabey D. Comparison of latex agglutination, wet preparation and culture for the detection of Trichomonas vaginalis. Sex Transm Infect 2004;80(3):201-203. ##Carney JA, Unadkat P, Yule A, Rajakumar R, Lacey CJ, Ackers JP. New rapid latex agglutination test for diagnosis of Trichomonas vaginalis infection. J Clin Pathol 1988;41(7):806-808. ##Darani HY, Doenhoff MJ. Anomalous immunogenic properties of serine proteases. Scand J Immunol 2009:70(4);384-388. ##Darani HY, Curtis RH, Mcneice C, Price HP, Sayers JR, Doenhoff MJ. Schistosoma mansoni: anomalous immunogenic properties of a 27 kDa larval serine protease associated with protective immunity. Parasitology 1997;115:237-247. ##Darani HY, Doenhoff MJ. An association between Schistosoma mansoni worms and an enzymatically-active protease/peptidase in mouse blood. Parasitology 2008;135:467-472. ##Hopwood V, Evans EGV, Carney JA. Rapid diagnosis of vaginal candidosis by latex particle agglutination. J Clin Pathol 1985;38:455-458. ##Lopez Abraham AM, Artiles Mde L, Ferandez Masso JR, San Martin DG, Alonso M, Alvarez RE. Evaluation of an agglutination method with latex particles sensitized for diagnosis of vaginal trichomoniasis. Rev Cubana Med Trop 2005;57(2):133-136. ##Fernandez Limia O, Lantero MI, Betancourt A, de Armas E, Villoch A. Prevalence of Candida albicans and Trichomonas vaginalis in pregnant women in Havana city by an immunologic latex agglutination test. MedGenMed. 2004;15;6(4):50. ##