Editorial 2 2 Genetically modified organisms (GMOs) have received a lot of attention in recent years, initially for their great potential in aiding the food shortage for a growing world population and later for the lower price, durability (resistance to insects, herbicides, viruses) and nutritional value of GM plants. In this issue of AJMB, a review article has addressed some of the major issues and concerns related to uses of GMOs. Due to recent progress in production of several GMOs by the agricultural biotechnology institutes in Iran, it is appropriate that issues such as potential health and environmental risks of GMOs are discussed in a biotechnology Journal such as AJMB. Since there are many issues that need to be addressed on the production and uses of GMOs, I have summarized some of the issues of concern for our readers and encourage submission of articles on this topic to AJMB. One of the major issues in GM foods is potential risks to human health. Safety assessment is an important method to address direct health effects (toxicity), allergenicity, nutritional or toxic properties, stability of the inserted gene, nutritional effects associated with genetic modification. Gene transfer and outcrossing are the two major concerns in the area of food safety and food security. If gene transfer (particularly antibiotic resistance genes) from GM foods to cells or bacteria in the gastrointestinal tract occurs, it would cause concern if the transferred genetic material adversely affects human health. The outcrossing can also occur if the movement of genes from GM plants into conventional crops or related species in the wild occurs. This problem has occurred when traces of a maize type only approved for feed use appeared in maize products for human consumption in the United States of America. Another major issue of concern is how GMO affects the environment? It is known that GMOs are capable to escape and potentially introduce the newly engineered genes into wild populations which then can potentially lead to loss of biodiversity on earth. Intellectual property rights and monopolization are also issues that are likely to become very significant as competition increases among nations and companies in applying biotechnology in improving food and medicine. The release of GMOs into the environment and the marketing of GM foods have in recent years resulted in public debates in many parts of the world, especially in the European Union. This debate is predicted to continue and intensify even more in the coming years mainly due to uses of biotechnology in medicine and its implications for human societies. It is appropriate that the leaders of biotechnology institutes and appropriate governmental bodies supervising the production and release of GMOs in Iran, actively participate in a national dialogue on the use and safety of GMOs. Furthermore, they should provide the right information to the public on matters of safety and security of GM foods produced domestically or imported to Iran from GM food producing countries. 108 108 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 175.pdf ####

Genetically Modified Foods and Social Concerns 2 2 Biotechnology is providing us with a wide range of options for how we can use agricultural and commercial forestry lands. The cultivation of genetically modified (GM) crops on millions of hectares of lands and their injection into our food chain is a huge global genetic experiment involving all living beings. Considering the fast pace of new advances in production of genetically modified crops, consumers, farmers and policymakers worldwide are challenged to reach a consensus on a clear vision for the future of world food supply. The current food biotechnology debate illustrates the serious conflict between two groups: 1) Agri-biotech investors and their affiliated scientists who consider agricultural biotechnology as a solution to food shortage, the scarcity of environmental resources and weeds and pests infestations; and 2) independent scientists, environmentalists, farmers and consumers who warn that genetically modified food introduces new risks to food security, the environment and human health such as loss of biodiversity; the emergence of superweeds and superpests; the increase of antibiotic resistance, food allergies and other unintended effects. This article reviews major viewpoints which are currently debated in the food biotechnology sector in the world. It also lays the groundwork for deep debate on benefits and risks of Biotech-crops for human health, ecosystems and biodiversity. In this context, although some regulations exist, there is a need for continuous vigilance for all countries involved in producing genetically engineered food to follow the international scientific biosafety testing guidelines containing reliable pre-release experiments and post-release track of transgenic plants to protect public health and avoid future environmental harm. 109 117 Behrokh Mohajer Maghari Biotechnology Department, Iranian Research for Science and Technology (IROST) Iran Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran FoodGenetically EngineeredGenetically modifiedGMOsHealthHumans 64.pdf Halford NG, Shewry PR. Genetically modified crops: methodology, benefits, regulation and public concerns. Br Med Bull 2000;56 (1):62-73.##Caiping MA, Stauss SH, Meilan R. Agrobacterium-mediated transformation of the genome-sequenced polar clone, nisqually-1 (Populus trichocarpa). Plant Mol Biol Rep 2004; 22:1-9.##Lee YS, Wetzel ED, Wagner NJ. The ballistic impact characteristics of Kevlar® woven fabrics impregnated with a colloidal shear thickening fluid. J Mater Sci 2003;38(13):2825-2833.##Obert B, Ponya Z, Pret’ova A, Barnabas B. Optimization of electroporation conditions for maize microspores. Maydica 2004;49:15-19. ##Darabani B, Farajnia S, Toorchi M, Zakerbostanabad S, Noeparvar S, Stewart N. DNA-delivery methods to produce transgenic plants. Biotechnology 2008;7(3):385-402.##Meli VS, Ghosh S, Prabha TN, Chakraborty N, Chakraborty S, Datta A. Enhancement of fruit shelf life by suppressing N-glycan processing enzymes. PNAS 2010;107(6):2413-2418. ##Consensus Document on Molecular Characterization of Plants Derives from Modern Biotechnology. Organization for Economic Co-operation and Development: Series on Harmonisation of Regulatory Oversight in Biotechnology No. 51 and Series on the Safety of Novel Foods and Feeds No. 2220. ENV/JM/MONO 2010; 41. Available from: http://www.oecd.org/dataoecd/16/29/46815346.pdf.##Knight B. Agricultural biotechnology in Europe. Crop Protection Monthly 2007 July.##Global Biotech Area Surges Past 100 Million Hectares on 13 Percent Growth: Interanational Service for the Acquisition of Agri. Biotech Applications ISAAA (US);2007 Jan. Available from: http://www.bionity.com/en/ news/61027/.##Brookes G, Barfoot P. GM crops The First Ten Years-Global Socio-economic and Environmental Impacts. PG Economics Ltd., UK: The International Service for the Acquisition of Agri-biotech Applications (ISAAA); 2006.##Rikki Stancich. GM food special report: Crops that survive climate change. Climate Change group; 2008 Feb. Available from: http://www.climatechangecorp.com/content.asp?ContentID=5157.##Improving the Regulation of Genetically Modified Foods and Other Novel Foods in Canada. Canadian Biotechnology Advisory Committee, Report to the Government of Canada Biotechnology Ministerial Coordinating Committee; 2002 Aug. Available from: http://dsp-psd.pwgsc.gc.ca/Collection/C2-589-2001-1E.pdf.##Altieri MA, Rosset P. Strengthening the case for why biotechnology will not help. The developing world: A response to MCGloughlin. AgBioForum 1999;2(3-4):226-236. ##Pusztai A. Genetically Modified Foods: Are They a Risk to Human/Animal Health? ActionBioscience; 2001 June. Available from: http://www.ask-force.org/web/Pusztai/Pusztai-GM-Foods-Risk-Human-Animal-Health-2001.PDF. ##GM Crops-the health effects. Soil Association. www.soilassociation.org. February (2008).##Mae-Wan Ho. Genetic Engineering Dream or Nightmare?: Turning the Tide on the Brave New World of Bad Science and Big Business. 2 Rev Upd edition. International Publishing Group Continuum; 2000.##Mae-Wan Ho. Stability of All Transgenic Lines in Doubt. ISIS Report. Institute of science in society; 2003 March. Available from: http://www.i-sis.org.uk/MON810GenomeRearranged.php.##Mae-Wan Ho. Transgenic Lines Unstable hence Illegal and Ineligible for Protection. ISIS Report. Institute of science in technology; 2008 March. Available from: http://www.i-sis.org.uk/transgenicLinesUnstable2.php.##Braun R. Antibiotic Resistance Markers in Genetically Modified (GM) Crops. European Federation of Biotechnology. Task Group On Public Perceptions of Biotechnology; 2001 Sep. Available from: http://www.biosafety.be/ARGMO/Documents/EFB_AntibioticRM_English.pdf.##Chen C, Thiruvengadam V, Lin W, Chang H, Hsu W. Lysine racemase: a novel non-antibiotic selectable marker for plant transformation. Plant Mol Biol 2010;72(1-2):153-169.##Non-Human Antimicrobial Usage and Antimicrobial Resistance: Scientific assessment. Joint FAO/OIE/WHO Expert Workshop: Geneva, 2003 Dec. Available from: http://www.who.int/foodsafety/publications/micro/en/amr.pdf.##Ho MW, Cummins J. New evidence links CaMV 35S promoter to HIV transcription. Microb Ecol Health Dis 2009;21(3-4):172-174. ##Myhre MR, Fenton KA, Eggert J, Nielsen KM, Traavik T. The 35S CaMV plant virus promoter is active in human enterocyte-like cells. Eur Food Res Technol 2006;222(1-2): 185-193.##Lai MM. RNA recombination in animal and plant viruses. Microbiol Mol Biol Rev 1992; 56(1):61-79.##Teycheney PY, Tepper M. Possible roles of endogenous plant viral sequences and transgenes containing viral sequences in both virus resistance and virus emergence. Environ Biosafety Res 2007;6(4):219-221.##Fares NH, El-Sayed AK. Fine structural changes in the ileum of mice fed on delta-endotoxin-treated potatoes and transgenic potatoes. Nat Toxins 1998;6(6):219-233. ##Ewen SWB, Pusztai A. Effects of diets containing genetically modified potatoes expressing Galanthus nivalis lectin on rat small intestine. The Lancet 1999;354(9187): 1353-1354.##Hashimoto W, Momma K, Katsube T, OhkawaY, IshigeT, Kito M, et al. Safety assessment of genetically engineered potatoes with designed soybean glycinin: compositional analyses of the potato tubers and digestibility of the newly expressed protein in transgenic potatoes. J Sci Food Agric 1999;79(12):1607-1612.##Momma K, Hashimoto W, Ozawa S, Kawai S, Katsube T, Takaiwa F, et al. Quality and safety evaluation of genetically engineered rice with soybean glycinin: Analyses of the grain composition and digestibility of glycinin in transgenic rice. Biosci Biotechnol Biochem 1999;63(2):314-318.##Velimirov A, Binter C, Zentek J. Biological effects of transgenic maize NK603xMON810 fed in long term reproduction studies in mice. Report by Institute fur Ernahrung, Austria. 2008 Nov. Available from: http://www.biosicherheit.de/pdf/aktuell/zentek_studie_2008.pdf.##Munro S. GM food debate. The Lancet 1999;354(9191):1727-1729. ##Daniel KT. The Hidden Dangers of Soy Allergens. Nexus Magazine 2004 Sep. Available from: www.nexusmagazine.com.##Ho MW, Cummins J. Agrobacterium & Morgellons Disease, A GM Connection? Global Research 2008 Aug. Available from: http://www.globalresearch.ca/index.php?context=va&aid=9891.##Kurunganti K. Mass Protests against GM Crops in India. Institute of science in technology. 2008 Apr. Available from: http://www.i-sis.org.uk/gmProtestsIndia.php.##Bernstein IL, Bernstein JA, Miller M, Tierzieva S, Bernstein DI, Lummus Z, et al. Immune responses in farm workers after exposure to Bacillus thuringiensis pesticides. Environ Health Perspect 1999;107(7):575-582. ##Vazquez-Padron RI, Moreno-Fierros L, Neri-Bazan L, Martinez-Gil AF, de la Riva, GA, Lopez-Revilla R. Characterization of the mucosal and systemic immune response induced by Cry1Ac protein from Bacillus thuringiensis HD 73 in mice. Braz J Med Biol Res 2000; 33(2):147-155. ##Pasini G, Simonato B, Curioni A, Vincenzi S, Cristaudo A, Santucci B, et al. IgE-mediated allergy to corn: a 50 kDa protein, belonging to the reduced soluble proteins, is a major allergen. Allergy 2002;57(2):98-106. ##Pusztai A. Can science give us the tools for recognizing possible health risks of GM foods. Nutr Health 2002;16:73-84. ##Millstone E, Brunner E, Mayer S. Beyond substantial equivalence. Nature 1999;401(6753) 525-526.##Redenbaugh K, Hatt W, Martineau B, Kramer M, Sheehy R, Sanders R, Houck C, Emlay D. A case study of the FLAVR SAVRTM tomato. In: Safety Assessment of Genetically Engineered Fruits and Vegetables. USA: CRC Press, Inc. Boca Raton; 1992.##Padgette SR, Taylor NB, Nida DL, Bailey MR, MacDonald J, Holden L, Fuchs RL. The composition of glyphosate-tolerant soybean seeds is equivalent to that of conventional soybeans. J Nutr 1996;126(3):702-716.##Taylor NB, Fuchs RL, MacDonald J, Shariff AB, Padgette SR. Compositional analysis of glyphosate-tolerant soybeans treated with glyphosate. J Agric Food Chem 1999; 47(10):4469-4473.##Berberich SA, Ream JE, Jackson TL, Wood R, Stipanovic R, Harvey P, et al. The composition of insect-protected cottonseed is equivalent to that of conventional cottonseed. J Agric Food Chem 1996;44(1):365-371. ##NovakWK,Haslberger AG. Substantial equivalence of antinutrients and inherent plant toxins in genetically modified novel foods. Food Chem Tox 2000;38(6):473-483. ##LappeMA, Bailey EB, Childress C,Setchell KDR. Alterations in clinically important phytoestrogens in genetically modified, herbicide-tolerant soybeans. J Med Food 1999; 1(4);241-245. ##Harrison LA, Bailey MR, Naylor MW, Ream JE, Hammond BG, Nida DL, et al. The expressed protein in glyphosate-tolerant soybean, 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4, is rapidly digested in vitro and is not toxic to acutely gavaged mice. J Nutr 1996;126(3):728-740.##Momma K, Hashimoto W, Ozawa S, Kawai S, Katsube T, Takaiwa F, et al. Quality and safety evaluation of genetically engineered rice with soybean glycinin: Analyses of the grain composition and digestibility of glycinin in transgenic rice. Biosci Biotechnol Biochem 1999;63(2):314-318.##Nordlee JA, Taylor SL, Townsend JA, Thomas LA, Bush RK. Identification of a Brazil nut allergen in transgenic soybean. N Engl J Med 1996;334:688-692. ##Bindslev-Jensen C, Poulsen LK. Hazards of unintentional/intentional introduction of allergens into foods. Allergy 1997;52(12):1184-1186. ##Burks AW, Fuchs RL. Assessment of the endogenous allergens in glyphosate-tolerant and commercial soybean varieties. J Allergy Clin Immunol 1995;96(6 pt 1):1008-1010. ##Nakamura R, Matsuda T. Rice allergenic protein and molecular-genetic approach for hypoallergenic rice. Biosci Biotech Biochem 1996;60(8):1215-1221. ##Carter CA, Gruere GP. International Approaches to the Labeling of Genetically Modified Foods. Agricultural Marketing Resource Center. University of California Department of Agricultural and Resource Economics 2003 Mar. Available from: http://www.agmrc.org/media/cms/cartergruere_929BEB69BA4EE.pdf.##Byrne P. Labeling of Genetically Engineered Foods. (2010). Colorado State University Extension. Fact sheet. No. 9.371. 2010 Sep. Available from: http://www.ext.colostate.edu/pubs/foodnut/09371.html.##Indepth: Genetic Modification, Genetically Modified Foods: a primer, CBC News Online. 2004 May. Available from: http://www.cbc.ca/news/background/genetics_modification/.##Most people want to know if their food has GM ingredients. Relax News. 2009 Dec. Available from: http://www.food.gov.uk.##Lendman S. Potential Health Hazards of Genetically Engineered Foods. 2008 Feb. Available from: www.truehealthfacts.com. ##Genetically modified foods. Better health Channel. 2010 Feb. Available from: www.betterhealth.vic.gov.au##Soybeans Monsanto. Available from: http://www.monsanto.com/products/Documents/pipeline-brochures/soybeans.pdf. ##Bourgeois F. Drug trials funded by industry are more likely to publish favorable results. Ann Int Med 2010;153:158-166.##Saunders P. Corporate Monopoly of Science. Institute of Science for Society. ISIS Report. 2009 Apr. Available from: http://www.i-sis.org.uk/corporateMonopolyOfScience.php.##Qaim M. Benefits of genetically modified crops for the poor: household income, nutrition, and health. New Biotechnol 2010;27(5):552-557.##Ho MW. Schmeiser’s Battle for the Seed. Institute of Science for Society. ISIS Report. Available from: http://www.i-sis.org.uk/SLBFTS.php.##

Comparative Expression Profile of Orphan Receptor Tyrosine Kinase ROR1 in Iranian Patients with Lymphoid and Myeloid Leukemias 2 2 It has recently been shown that ROR1, a member of the receptor tyrosine kinase family, is overexpressed in leukemic B cells of Chronic Lymphocytic Leukemia (CLL) and a subset of Acute Lymphoblastic Leukemia (ALL). In this comparative study the expression profile of ROR1 mRNA was investigated in Iranian patients with CLL and Acute Myelogenous Leukemia (AML) and the results were compared with those previously reported in our Iranian ALL patients. RT-PCR was performed on bone marrow and/or peripheral blood samples of 84 CLL and 12 AML patients. CLL samples were classified into immunoglobulin heavy chain variable region (IGHV) gene mutated (n=55) and unmutated (n=29) and also indolent (n=42) and progressive (n=39) subtypes. ROR1 expression was identified in 94% of our CLL patients, but none of the AML patients expressed ROR1. No significant differences were observed between different CLL subtypes for ROR1 expression. Taken together the present data and our previous results on ROR1 expression in ALL, our findings propose ROR1 as a tumor-associated antigen overexpressed in a large proportion of lymphoid (CLL and ALL), but not myeloid (AML) leukemias. Expression of ROR1 seems to be associated to lineage and differentiation stages of leukemic cells with a potential implication for immunotherapy. 119 125 Mahdi Shabani Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Public Health, Tehran University of Medical Sciences Iran Hossein Asgarian-Omran Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Public Health, Tehran University of Medical Sciences Iran Mohammad Hojjat-Farsangi Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Public Health, Tehran University of Medical Sciences Iran Parvaneh Vossough Clinic of Hematology, Ali-Asghar Hospital, Faculty of Medicine, Iran University of Medical Sciences Clinic of Hematology, Ali-Asghar Hospital, Faculty of Medicine, Iran University of Medical Sciences Iran Ramazan A.Sharifian Clinic of Hematology and Oncology, Vali-Asr Hospital, Faculty of Medicine, Tehran University of Medical Sciences Clinic of Hematology and Oncology, Vali-Asr Hospital, Faculty of Medicine, Tehran University of Medical Sciences Iran Gholam Reza Toughe Clinic of Hematology and Oncology, Vali-Asr Hospital, Faculty of Medicine, Tehran University of Medical Sciences Clinic of Hematology and Oncology, Vali-Asr Hospital, Faculty of Medicine, Tehran University of Medical Sciences Iran Seyed Mohsen Razavi Clinic of Hematology and Oncology, Firozgar Hospital, Faculty of Medicine, Iran University of Medical Sciences Clinic of Hematology and Oncology, Firozgar Hospital, Faculty of Medicine, Iran University of Medical Sciences Iran Jalal Khoshnoodi Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Public Health, Tehran University of Medical Sciences Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Hodjattallah Rabbani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECRImmune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska Hospital, Karolinska Institutet Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR IranSweden Fazel Shokri Clinic of Hematology, Ali-Asghar Hospital, Faculty of Medicine, Iran University of Medical SciencesMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR Clinic of Hematology, Ali-Asghar Hospital, Faculty of Medicine, Iran University of Medical SciencesMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR IranIran Acute myelocytic leukemiaChronic lymphocytic leukemiaROR1RT-PCR 65.pdf Masiakowski P, Carroll RD. A novel family of cell surface receptors with tyrosine kinase-like domain. J Biol Chem 1992;267(36):26181-26190.##Nomi M, Oishi I, Kani S, Suzuki H, Matsuda T, Yoda A, et al. Loss of mRor1 enhances the heart and skeletal abnormalities in mRor2-deficient mice: redundant and pleiotropic functions of mRor1 and mRor2 receptor tyrosine kinases. Mol Cell Biol 2001;21(24):8329-8335.##Wang HY, Liu T, Malbon CC. Structure-function analysis of Frizzleds. Cell Signal 2006;18(7):934-941.##Grumolato L, Liu G, Mong P, Mudbhary R, Biswas R, Arroyave R, et al. Canonical and noncanonical Wnts use a common mechanism to activate completely unrelated coreceptors. Genes Dev 2010;24(22):2517-2530.##Fukuda T, Chen L, Endo T, Tang L, Lu D, Castro JE, et al. Antisera induced by infusions of autologous Ad-CD154-leukemia B cells identify ROR1 as an oncofetal antigen and receptor for Wnt5a. Proc Natl Acad Sci USA 2008;105 (8):3047-3052.##Klein U, Tu Y, Stolovitzky GA, Mattioli M, Cattoretti G, Husson H, et al. Gene expression profiling of B cell chronic lymphocytic leukemia reveals a homogeneous phenotype related to memory B cells. J Exp Med 2001;194(11):1625-1638.##Rosenwald A, Alizadeh AA, Widhopf G, Simon R, Davis RE, Yu X, et al. Relation of gene expression phenotype to immunoglobulin mutation genotype in B cell chronic lymphocytic leukemia. J Exp Med 2001;194(11):1639-1647.##Haiat S, Billard C, Quiney C, Ajchenbaum-Cymbalista F, Kolb JP. Role of BAFF and APRIL in human B-cell chronic lymphocytic leukaemia. Immunology 2006;118(3):281-292.##Dighiero G, Hamblin TJ. Chronic lymphocytic leukaemia. Lancet 2008;371(9617):1017-1029.##Hamblin TJ. Prognostic markers in chronic lymphocytic leukaemia. Best Pract Res Clin Haematol 2007;20(3):455-68.##Baskar S, Kwong KY, Hofer T, Levy JM, Kennedy MG, Lee E, et al. Unique cell surface expression of receptor tyrosine kinase ROR1 in human B-cell chronic lymphocytic leukemia. Clin Cancer Res 2008;14(2):396-404.##Hojjat-Farsangi M, Jeddi-Tehrani M, Razavi SM, Sharifian RA, Mellstedt H, Shokri F, et al. Immunoglobulin heavy chain variable region gene usage and mutational status of the leukemic B cells in Iranian patients with chronic lymphocytic leukemia. Cancer Science 2009;100(12):2346-2353.##Daneshmanesh AH, Mikaelsson E, Jeddi-Tehrani M, Bayat AA, Ghods R, Ostadkarampour M, et al. Ror1, a cell surface receptor tyrosine kinase is expressed in chronic lymphocytic leukemia and may serve as a putative target for therapy. Int J Cancer 2008;123(5):1190-1195.##Rabbani H, Ostadkarampour M, Daneshmanesh AH, Basiri A, Jeddi-Tehrani M, Forouzesh F. Expression of ROR1 in patients with renal cancer--a potential diagnostic marker. Iran Biomed J 2010;14(3):77-82.##Hudecek M, Schmitt TM, Baskar S, Lupo-Stanghellini MT, Nishida T, Yamamoto TN, et al. The B-cell tumor-associated antigen ROR1 can be targeted with T cells modified to express a ROR1-specific chimeric antigen receptor. Blood 2010;116(22):4532-4541.##Barna G, Mihalik R, Timar B, Tombol J, Csende Z, Sebestyen A, et al. ROR1 expression is not a unique marker of CLL. Hematol Oncol 2011;29(1):17-21. ##Shabani M, Asgarian-Omran H, Jeddi-Tehrani M, Vossough P, Faranoush M, Sharifian RA, et al. Overexpression of orphan receptor tyrosine kinase Ror1 as a putative tumor-associated antigen in Iranian patients with acute lymphoblastic leukemia. Tumour Biol 2007;28(6):318-326.##Hamblin TJ, Davis Z, Gardiner A, Oscier DG, Stevenson FK. Unmutated Ig V(H) genes are associated with a more aggressive form of chronic lymphocytic leukemia. Blood 1999;94(6):1848-1854.##Kazemi T, Asgarian-Omran H, Hojjat-Farsangi M, Shabani M, Memarian A, Sharifian RA, et al. Fc receptor-like 1-5 molecules are similarly expressed in progressive and indolent clinical subtypes of B-cell chronic lymphocytic leukemia. Int J Cancer 2008;123(9):2113-2119.##Asgarian Omran H, Shabani M, Shahrestani T, Sarafnejad A, Khoshnoodi J, Vossough P, et al. Over-expression of Wilm’s Tumor Gene 1(WT1) in Iranian patients with acute myeloblastic leukemia. Iran J Immunol 2005;2(4):182-190.##Farsangi MH, Jeddi-Tehrani M, Sharifian RA, Razavi SM, Khoshnoodi J, Rabbani H, et al. Analysis of the immunoglobulin heavy chain variable region gene expression in Iranian patients with chronic lymphocytic leukemia. Leuk Lymphoma 2007;48(1):109-116.##Greaves MF. Biological models for leukaemia and lymphoma. IARC Sci Publ 2004(157):351-372.##Thiel E. Cell surface markers in leukemia: biological and clinical correlations. Crit Rev Oncol Hematol 1985;2(3):209-260.##Dunphy CH. Gene expression profiling data in lymphoma and leukemia: review of the literature and extrapolation of pertinent clinical applications. Arch Pathol Lab Med 2006;130(4):483-520.##Virtaneva K, Wright FA, Tanner SM, Yuan B, Lemon WJ, Caligiuri MA, et al. Expression profiling reveals fundamental biological differences in acute myeloid leukemia with isolated trisomy 8 and normal cytogenetics. Proc Natl Acad Sci USA 2001;98(3):1124-1129.##Wang J, Coombes KR, Highsmith WE, Keating MJ, Abruzzo LV. Differences in gene expression between B-cell chronic lymphocytic leukemia and normal B cells: a meta-analysis of three microarray studies. Bioinformatics 2004;20(17):3166-3178.##Scrideli CA, Cazzaniga G, Fazio G, Pirola L, Callegaro A, Bassan R, et al. Gene expression profile unravels significant differences between childhood and adult Ph+ acute lymphoblastic leukemia. Leukemia 2003;17(11):2234-2237.##Alizadeh AA, Eisen MB, Davis RE, Ma C, Lossos IS, Rosenwald A, et al. Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. Nature 2000;403(6769):503-511.##Basso K, Liso A, Tiacci E, Benedetti R, Pulsoni A, Foa R, et al. Gene expression profiling of hairy cell leukemia reveals a phenotype related to memory B cells with altered expression of chemokine and adhesion receptors. J Exp Med 2004;199(1):59-68.##Mikaelsson E, Daneshmanesh AH, Luppert A, Jeddi-Tehrani M, Rezvany MR, Sharifian RA, et al. Fibromodulin, an extracellular matrix protein: characterization of its unique gene and protein expression in B-cell chronic lymphocytic leukemia and mantle cell lymphoma. Blood 2005;105(12):4828-4835.##Memarian A, Hojjat-Farsangi M, Asgarian-Omran H, Younesi V, Jeddi-Tehrani M, Sharifian RA, et al. Variation in WNT genes expression in different subtypes of chronic lymphocytic leukemia. Leuk Lymphoma 2009;50(12):2061-2070.##Yoda A, Oishi I, Minami Y. Expression and function of the Ror-family receptor tyrosine kinases during development: lessons from genetic analyses of nematodes, mice, and humans. J Recept Signal Transduct Res 2003;23(1):1-15.##Li P, Harris D, Liu Z, Liu J, Keating M, Estrov Z. Stat3 activates the receptor tyrosine kinase like orphan receptor-1 gene in chronic lymphocytic leukemia cells. PloS One 2010;5(7):e11859.##MacKeigan JP, Murphy LO, Blenis J. Sensitized RNAi screen of human kinases and phosphatases identifies new regulators of apoptosis and chemoresistance. Nat Cell Biol 2005;7(6):591-600.##Muller-Tidow C, Schwable J, Steffen B, Tidow N, Brandt B, Becker K, et al. High-throughput analysis of genome-wide receptor tyrosine kinase expression in human cancers identifies potential novel drug targets. Clin Cancer Res 2004;10(4):1241-1249.##

Expression and Purification of Functionally Active Recombinant Human Alpha 1-Antitrypsin in Methylotrophic Yeast Pichia Pastoris 2 2 Human alpha 1-antitrypsin (AAT) cDNA was obtained from HepG2 cell lines. After PCR and construction of expression vector pPICZα-AAT, human AAT was expressed in the yeast Pichia pastoris (P.pastoris) in a secretary manner and under the control of inducible alcohol oxidase 1 (AOX1) promoter. The amount of AAT protein in medium was measured as 60 mg/l 72 hr after induction with methanol. Results indicated the presence of protease inhibitory function of the protein against elastase. Purification was done using His-tag affinity chromatography. Due to the different patterns of glycosylation in yeast and human, the recombinant AAT showed different SDS-PAGE patterns compared to that of serum-derived AAT while pI shifted from 4.9 in native AAT compared to 5.2 in recombinant AAT constructed in this study. 127 134 Sareh Arjmand Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares UniversityNational Institute of Genetic Engineering and Biotechnology (NIGEB)School of Pharmacy, Zanjan University of Medical Science Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares UniversitySchool of Pharmacy, Zanjan University of Medical Science IranIranIran Elham Bidram Department of Clinical Biochemistry, Tarbiat Modares University Department of Clinical Biochemistry, Tarbiat Modares University Iran Abbas Sahebghadam Lotfi National Institute of Genetic Engineering and Biotechnology (NIGEB)Department of Clinical Biochemistry, Tarbiat Modares University Department of Clinical Biochemistry, Tarbiat Modares University IranIran Mehdi Shamsara National Institute of Genetic Engineering and Biotechnology (NIGEB) Iran Seyed Javad Mowla Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University Iran Alpha 1-antitrypsinPichia PastorisProtease inhibitorsRecombinant proteins 66.pdf Crowther DC, Belorgey D, Miranda E, Kinghorn KJ, Sharp LK, Lomas DA. Practical genetics: alpha-1-antitrypsin deficiency and the serpinopathies. Eur J Hum Genet 2004;12(3):167-172.##Crystal RG. The alpha 1-antitrypsin gene and its deficiency states. Trends Genet 1989;5(12):411-417.##Nita I, Hollander C, Westin U, Janciauskiene SM. Prolastin, a pharmaceutical preparation of purified human alpha1-antitrypsin, blocks endotoxin-mediated cytokine release. Respir Res 2005;3(1):12-18.##Wewers MD, Casolaro MA, Sellers SE, Swayze SC, McPhaul KM, Wittes JT, et al. Replacement therapy for alpha 1-antitrypsin deficiency associated with emphysema. N Engl J Med 1987;316(17):1055-1062.##Tirado-Conde G, Lara B, Miravitlles M. Augmentation therapy for emphysema due to alpha-1-antitrypsin deficiency. Ther Adv Respir Dis 2008;2(1):13-21.##Stocks JM, Brantly ML, Wang-Smith L, Campos MA, Chapman KR, Kueppers F, et al. Pharmacokinetic comparability of Prolastin(R)-C to Prolastin(R) in alpha-antitrypsin deficiency: a randomized study. BMC Clin Pharmacol 2010;10:13-24.##Dodd RY, Leiby DA. Emerging infectious threats to the blood supply. Annu Rev Med 2004;55:191-207.##Karnaukhova E, Ophir Y, Golding B. Recombinant human alpha-1 proteinase inhibitor: towards therapeutic use. Amino Acids 2006;30(4):317-332.##Kwon KS, Song M, Yu MH. Purification and characterization of alpha 1-antitrypsin secreted by recombinant yeast Saccharomyces diastaticus. J Biotechnol 1995;42(3):191-195.##Massoud M, Bischoff R, Dalemans W, Pointu H, Attal J, Schultz H, et al. Expression of active recombinant human alpha 1-antitrypsin in transgenic rabbits. J Biotechnol 1991;18(3):193-203.##Macauley-Patrick S, Fazenda ML, McNeil B, Harvey LM. Heterologous protein production using the Pichia pastoris expression system. Yeast 2005;22(4):249-270.##Ghaedi M, Lotfi AS, Soleimani M, Shamsara M, Arjmand S, Adibi B. Expression of recombinant alpha-1 antitrypsin in CHO and COS-7 cell line using antiviral vector. Iranian J Biotechnol 2009;7(3):148-156.##Cregg JM. Introduction: distinctions between Pichia pastoris and other expression systems. Methods Mol Biol 2007;389:1-10.##Bretthauer RK, Castellino FJ. Glycosylation of Pichia pastoris-derived proteins. Biotechnol Appl Biochem 1999;30( Pt 3):193-200.##Mattanovich D, Graf A, Stadlmann J, Dragosits M, Redl A, Maurer M, et al. Genome, secretome and glucose transport highlight unique features of the protein production host Pichia pastoris. Microb Cell Fact 2009;8:29-42.##Celis JE, Carter N, Hunter T, Simons K, Small JV, Shotton D, (eds). Cell biology: A Laboratory Handbook. San Diego: Elsevier, Academic Press; 2006,219-224.##Cos O, Ramon R, Montesinos JL, Valero F. Operational strategies, monitoring and control of heterologous protein production in the methylotrophic yeast Pichia pastoris under different promoters: a review. Microb Cell Fact 2006;5:17-37.##Senerovic L, Stankovic N, Spizzo P, Basso A, Gardossi L, Vasiljevic B, et al. High-level production and covalent immobilization of Providencia rettgeri penicillin G acylase (PAC) from recombinant Pichia pastoris for the development of a novel and stable biocatalyst of industrial applicability. Biotechnol Bioeng 2006;93(2):344-354.##Cregg JM, Vedvick TS, Raschke WC. Recent advances in the expression of foreign genes in Pichia pastoris. Biotechnology 1993;11(8):905-910.##Morton CL, Potter PM. Comparison of Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, Spodoptera frugiperda, and COS7 cells for recombinant gene expression. Application to a rabbit liver carboxylesterase. Mol Biotechnol 2000;16(3):193-202.##Powell LM, Pain RH. Effects of glycosylation on the folding and stability of human, recombinant and cleaved alpha 1-antitrypsin. J Mol Biol 1992;224(1):241-52.##Kwon KS, Yu MH. Effect of glycosylation on the stability of alpha1-antitrypsin toward urea denaturation and thermal deactivation. Biochim Biophys Acta 1997;1335(3):265-272.##Kang HA, Sohn JH, Choi ES, Chung BH, Yu MH, Rhee SK. Glycosylation of human alpha 1-antitrypsin in Saccharomyces cerevisiae and methylotrophic yeasts. Yeast 1998;14(4):371-381.##Trimble RB, Atkinson PH, Tschopp JF, Townsend RR, Maley F. Structure of oligosaccharides on Saccharomyces SUC2 invertase secreted by the methylotrophic yeast Pichia pastoris. J Biol Chem 1991;266(34):22807-22817.##Renversez JC, Lebrun-Fourcy C, Rondot J, Revol C. Determination of alpha 1 antitrypsin phenotypes in plasma using isoelectric focusing on this agarose gel. Pathol Biol 1998;46(1):15-20.##Zerimech F, Hennache G, Bellon F, Barouh G, Jacques Lafitte J, Porchet N, et al. Evaluation of a new Sebia isoelectrofocusing kit for alpha 1-antitrypsin phenotyping with the Hydrasys system. Clin Chem Lab Med 2008;46(2):260-263.##

Construction and In vitro Expression Analyses of a DNA Plasmid Encoding Dense Granule GRA5 Antigen of Toxoplasma gondii 2 2 Toxoplasmosis is an infection caused by the protozoan parasite Toxoplasma gondii (T.gondii) throughout the world. Although usually asymptomatic, the infection can cause serious medical problems in immunocompromised individuals and fetuses. Toxoplasmosis also causes considerable economic loss because of abortion in livestock. DNA vaccination is a promising approach against intracellular parasites such as T.gondii. The goal of this study was to construct and evaluate functionality of a mammalian plasmid expressing GRA5 anti-gen of T.gondii as a possible DNA vaccine. GRA5 gene fragment, devoid of the signal sequence, was amplified from genomic DNA of T.gondii RH strain, and cloned into pcDNA3.1 plasmid. The pcDNA3.1-GRA5 (pGRA5) was analyzed by restriction enzyme digestion followed by sequence determination. The pGRA5 was transfected into HEK 239-T human kidney cells, and expression of GRA5 antigen was investigated by Western blotting and immunofluorescence staining. The sequence encoding GRA5 was cloned into pcDNA3.1 plasmid. Restriction digestion of pGRA5 with Pst I enzyme showed correct insertion of GRA5 DNA into the plasmid. Sequence analysis revealed 100% homology with the published sequence of gra5. Immunofluorescence and Western blotting analyses of HEK 293-T cells transfected with pGRA5 showed specific expression of GRA5. Immunogenicity of pGRA5 will be evaluated in mice. 135 141 Jalal Babaie Molecular Parasitology Laboratory, Pasteur Institute of Iran Iran Ghazaleh Sadeghiani Molecular Parasitology Laboratory, Pasteur Institute of Iran Iran Majid Golkar Molecular Parasitology Laboratory, Pasteur Institute of Iran Iran Toxoplasma gondiiDNA vaccineGRA5 protein 67.pdf Kean BH. Clinical Toxoplasmosis-50 years. Trans R Soc Trop Med Hyg 1972;66(4):549-571.##Montoya JG, Liesenfeld O. Toxoplasmosis. Lancet 2004;363(9425):1965-1976.##Montoya JG, Remington JS. Management of Toxoplasma gondii infection during pregnancy. Clin Infect Dis 2008;47(4):554-566.##Unoki M, Brunet J, Mousli M. Drug discovery targeting epigenetic codes: the great potential of UHRF1, which links DNA methylation and histone modifications, as a drug target in cancers and toxoplasmosis. Biochem Pharmacol 2009;78(10):1279-1288.##Mamidi A, DeSimone JA, Pomerantz RJ. Central nervous system infections in individuals with HIV-1 infection. J Neurovirol 2002;8(3):158-167.##Dubey JP. Long-term persistence of Toxoplasma gondii in tissues of pigs inoculated with T gondii oocysts and effect of freezing on viability of tissue cysts in pork. Am J Vet Res 1988;49(6):910-913.##Buxton D, Thomson K, Maley S, Wright S, Bos HJ. Vaccination of sheep with a live incomplete strain (S48) of Toxoplasma gondii and their immunity to challenge when pregnant. Vet Rec 1991;129(5):89-93.##Mevelec MN, Ducournau C, Bassuny IA, Olivier M, Seche E, Lebrun M, et al. Mic1-3 Knockout Toxoplasma gondii is a good candidate for a vaccine against T. gondii-induced abortion in sheep. Vet Res 2010;41(4):49.##Kur J, Holec-Gasior L, Hiszczynska-Sawicka E. Current status of toxoplasmosis vaccine development. Expert Rev Vaccines 2009;8(6):791-808.##Jongert E, Roberts CW, Gargano N, Forster-Wald E, Petersen E. Vaccines against Toxoplasma gondii: challenges and opportunities. Mem Inst Oswaldo Cruz 2009;104(2):252-266.##Carvalho JA, Rodgers J, Atouguia J, Prazeres DM, Monteiro GA. DNA vaccines: a rational design against parasitic diseases. Expert Rev Vaccines 2010;9(2):175-191.##Liu MA. DNA vaccines: a review. J InternMed 2003;253(4):402-410.##Liu MA, Wahren B, Karlsson Hedestam GB. DNA vaccines: recent developments and future possibilities. Hum Gene Ther 2006;17(11):1051-1061.##Liu MA. Gene-based vaccines: Recent developments. Curr Opin Mol Ther 2010;12(1):86-93.##Angus CW, Klivington-Evans D, Dubey JP, Kovacs JA. Immunization with a DNA plasmid encoding the SAG1 (P30) protein of Toxoplasma gondii is immunogenic and protective in rodents. J Infect Dis 2000;181(1):317-324.##Nielsen HV, Di CM, Beghetto E, Spadoni A, Petersen E, Gargano N. Toxoplasma gondii: DNA vaccination with bradyzoite antigens induces protective immunity in mice against oral infection with parasite cysts. Exp Parasitol 2006;112(4):274-279.##Jongert E, De CS, Dewit J, Huygen K. GRA7 provides protective immunity in cocktail DNA vaccines against Toxoplasma gondii. Parasite Immunol 2007;29(9):445-453.##Desolme B, Mevelec MN, Buzoni-Gatel D, Bout D. Induction of protective immunity against toxoplasmosis in mice by DNA immunization with a plasmid encoding Toxoplasma gondii GRA4 gene. Vaccine 2000;18(23):2512-2521.##Cesbron-Delauw MF, Gendrin C, Travier L, Ruffiot P, Mercier C. Apicomplexa in mammalian cells: trafficking to the parasitophorous vacuole. Traffic 2008;9(5):657-664.##Mercier C, Adjogble KD, Daubener W, Delauw MF. Dense granules: are they key organelles to help understand the parasitophorous vacuole of all apicomplexa parasites? Int J Parasitol 2005;35(8):829-849.##Bivas-Benita M, Laloup M, Versteyhe S, Dewit J, De BJ, Jongert E, et al. Generation of Toxoplasma gondii GRA1 protein and DNA vaccine loaded chitosan particles: preparation, characterization, and preliminary in vivo studies. Int J Pharm 2003;266(1-2):17-27.##Martin V, Supanitsky A, Echeverria PC, Litwin S, Tanos T, De Roodt AR, et al. Recombinant GRA4 or ROP2 protein combined with alum or the gra4 gene provides partial protection in chronic murine models of toxoplasmosis. Clin Diagn Lab Immunol 2004;11(4):704-710.##Mevelec MN, Bout D, Desolme B, Marchand H, Magne R, Bruneel O, et al. Evaluation of protective effect of DNA vaccination with genes encoding antigens GRA4 and SAG1 associated with GM-CSF plasmid, against acute, chronical and congenital toxoplasmosis in mice. Vaccine 2005;23(36):4489-4499.##Scorza T, D'Souza S, Laloup M, Dewit J, De BJ, Verschueren H, et al. A GRA1 DNA vaccine primes cytolytic CD8(+) T cells to control acute Toxoplasma gondii infection. Infect Immun 2003;71(1):309-316.##Lecordier L, Mercier C, Sibley LD, Cesbron-Delauw MF. Transmembrane insertion of the Toxoplasma gondii GRA5 protein occurs after soluble secretion into the host cell. Mol Biol Cell 1999;10(4):1277-1287.##Lecordier L, Mercier C, Torpier G, Tourvieille B, Darcy F, Liu JL, et al. Molecular structure of a Toxoplasma gondii dense granule antigen (GRA 5) associated with the parasitophorous vacuole membrane. Mol Biochem Parasitol 1993;59(1):143-153.##Holec-Gasior L, Kur J. Toxoplasma gondii: Recombinant GRA5 antigen for detection of immunoglobulin G antibodies using enzyme-linked immunosorbent assay. Exp Parasitol 2010;124(3):272-278.##Zenner L, Estaquier J, Darcy F, Maes P, Capron A, Cesbron-Delauw MF. Protective immunity in the rat model of congenital toxoplasmosis and the potential of excreted-secreted antigens as vaccine components. Parasite Immunol 1999;21(5):261-272.##Igarashi M, Kano F, Tamekuni K, Machado RZ, Navarro IT, Vidotto O, et al. Toxoplasma gondii: evaluation of an intranasal vaccine using recombinant proteins against brain cyst formation in BALB/c mice. Exp Parasitol 2008;118(3):386-392.##Charif H, Darcy F, Torpier G, Cesbron-Delauw MF, Capron A. Toxoplasma gondii: characterization and localization of antigens secreted from tachyzoites. Exp Parasitol 1990;71(1):114-124.##Grangeot-Keros L, Mayaux MJ, Lebon P, Freymuth F, Eugene G, Stricker R, et al. Value of cytomegalovirus (CMV) IgG avidity index for the diagnosis of primary CMV infection in pregnant women. J Infect Dis 1997;175(4):944-946.##Kozak M. Regulation of translation via mRNA structure in prokaryotes and eukaryotes. Gene 2005;361:13-37.##Gendrin C, Mercier C, Braun L, Musset K, Dubremetz JF, Cesbron-Delauw MF. Toxoplasma gondii uses unusual sorting mechanisms to deliver transmembrane proteins into the host-cell vacuole. Traffic 2008;9(10):1665-1680.##

Biogenic Silver Nanoparticles by Gelidiella acerosa Extract and their Antifungal Effects 2 2 The synthesis, characterization and application of biologically synthesized nanomaterials are an important aspect in nanotechnology. The present study deals with the synthesis of silver nanoparticles (Ag-NPs) using the aqueous extract of red seaweed Gelidiella acerosa as the reducing agent to study the antifungal activity. The formation of Ag-NPs was confirmed by UV-Visible Spectroscopy, X-Ray Diffraction (XRD) pattern, Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The synthesized Ag-NPs was predominately spherical in shape and polydispersed. Fourier Transform Infra-Red (FT-IR) spectroscopy analysis showed that the synthesized nano-Ag was capped with bimolecular compounds which are responsible for reduction of silver ions. The antifungal effects of these nanoparticles were studied against Humicola insolens (MTCC 4520), Fusarium dimerum (MTCC 6583), Mucor indicus (MTCC 3318) and Trichoderma reesei (MTCC 3929). The present study indicates that Ag-NPs have considerable antifungal activity in comparison with standard antifungal drug, and hence further investigation for clinical applications is necessary. 143 148 Marimuthu Vivek Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore India Palanisamy Senthil Kumar Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore India Sesurajan Steffi Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore India Sellappa Sudha Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore India Gelidiella acerosaSilver nanoparticleNanotechnologySeaweed 68.pdf Balantrapu K, Goia D. Silver nanoparticles for printable electronics and biological applications. J Mater Res 2009;24(9):2828-2836.##Tripathi RM, Saxena A, Gupta N, Kapoor H, Singh RP. High antibacterial activity of silver nanoballs against E.coli MTCC 1302, S.typhimurium MTCC 1254, B.subtilis MTCC 1133 and P.aeruginosa MTCC 2295. Dig J Nanomater Bios 2010;5(2):323-330.##Patakfalvi R, Dekany I. Preparation of silver nanoparticles in liquid crystalline systems. Colloid Polym Sci 2010;280(5):461-470.##Rodriguez-Sanchez L, Blanco MC, Lopez-Quintela MA. Electrochemical synthesis of silver nanoparticles. J Phys Chem B 2000;104:9683-9688.##Taleb A, Petit C, Pileni MP. Optical properties of self-assembled 2D and 3D superlattices of silver nanoparticles. J Phys Chem B 1998;102(12):2214-2220.##Mondal S, Roy N, Laskar RA, Sk I, Basu S, Mandal D, et al. Biogenic synthesis of Ag, Au and bimetallic Au/Ag alloy nanoparticles using aqueous extract of Mahogany (Swietenia mahogani JACQ.) leaves. Colloids Surf B: Biointerfaces 2011;82(2):497-504.##Begum NA, Mondal S, Basu S, Laskar RA, Mandal D. Biogenic synthesis of Au and Ag nanoparticles using aqueous solutions of black tea leaf extracts. Colloids Surf B Biointerfaces 2009;71(1):113-118.##Kattumuri V, Katti K, Bhaskaran S, Boote EJ, Casteel SW, Fent GM, et al. Gum arabic as a phytochemical construct for the stabilization of gold nanoparticles: in vivo pharmacokinetics and X-ray-contrast-imaging studies. Small 2007;3(2):333-341.##Song JY, Kim BS. Biological synthesis of bimetallic Au/Ag nanoparticles using Persimmon (Diopyros kaki ) leaf extract. Korean J Chem Eng 2008;25(4):808-811.##Gilaki M. Biosynthesis of Silver nanoparticles using plant extracts. J Biol Sci 2010;10(5):465-467.##Govindaraju K, Kiruthiga V, Ganesh Kumar V, Singaravelu G. Extracellular synthesis of silver nanoparticles by a marine alga, Sargassum wightii Grevilli and their antibacterial effects. J Nanosci Nanotechnol 2009;9:5497-5501.##Premakumara GA, Ratnasooriya WD, Tillekeratne LM. Studies on the post-coital contraceptive mechanisms of crude extract of Sri Lankan marine red algae, Gelidiella acerosa. Contraception 1995;52(3):203-207.##Boulch F, Schouler MC, Donnadieu P, Chaix JM, Djurado E. Domain size distribution of Y-TZP nanoparticles using XRD and HRTEM. Image Anal Stereol 2001;20:157-161.##Sastry M, Patil V, Sainkar SR. Electrostatically controlled diffusion of carboxylic acid derivatized silver colloidal particles in thermally evaporated fatty amine films. J Phys Chem B 1998;102:1404-1410.##Mulvaney P. Surface plasmon spectroscopy of nanosized metal particles. Langmuir 1996;12(3):788-800.##Mukherjee P, Ahmad A, Mandal D, Senapati S, Sainkar SR, Khan MI, et al. Fungus-mediated synthesis of silver nanoparticles and their immobilization in the mycelial matrix: A novel biological approach to nanoparticle synthesis. Nano Lett 2001;1(10):515-519.##Mukherjee P, Senapati S, Mandal D, Ahmad A, Khan MI, Kumar R, et al. Extracellular synthesis of gold nanoparticles by the fungus Fusarium oxysporum. Chem Biochem 2002;3(5):461-463.##GonzaloJ, Serna R, Sol J, Babonneau D, Afonso CN. Morphological and interaction effects on the surface plasmon resonance of metal nanoparticles. J Phys Condens Matter 2003;15(42):3001-3002. ##Panacek A, Kolar M, Vecerova R, Prucek R, Soukupova J, Krystof V, et al. Antifungal activity of silver nanoparticles against Candida spp. Biomaterials 2009;30(31):6333-6340.##He L, Liu Y, Mustapha A, Lin M. Antifungal activity of zinc oxide nanoparticles against Botrytis cinerea and Penicillium expansum. Microbiol Res 2011;166(3):207-215. ##Klasen HJ. A historical review of the use of silver in the treatment of burns. II. Renewed interest for silver. Burns 2000;26(2):131-138.##Singh A, Jain D, Upadhyay MK, Khandelwal N, Vermahn. Green synthesis of silver nanoparticles using Argemone mexicana leaf extract and evaluation of their antimicrobial activities. Dig J Nanomater Bios 2010;5(2):483-489.##

Differentiation of Bovine Spermatogonial Stem Cells into Osteoblasts 2 2 Spermatogonial Stem Cell (SSC) technologies provide multiple opportunities for research in the field of biotechnology and regenerative medicine. The therapeutic use of Embryonic Stem Cells (ESCs) is restricted due to severe ethical and immunological concerns. Therefore, we need a new pluripotent cell type. Despite well-known role of germ cells in the gametogenesis, some facts apparently show their multipotentiality. In the present study, bovine SSCs were co-cultured with Sertoli cell for 7 days. Sertoli cells and SSCs were identified by Vimentin and Oct-4 immunocytochemical staining method, respectively. In order to differentiate SSCs into osteoblasts, we used consecutive inducer media without separation of the colonies. We characterized osteoblasts using Alizarin red staining. 149 153 Babak Qasemi-Panahi Department of Clinical Science, Faculty of Veterinary Medicine, University of Tehran Department of Clinical Science, Faculty of Veterinary Medicine, University of Tehran Iran Parviz Tajik Department of Clinical Science, Faculty of Veterinary Medicine, University of Tehran Department of Clinical Science, Faculty of Veterinary Medicine, University of Tehran Iran Mansooreh Movahedian Department of Anatomy, Faculty of Medical Science, University of Tarbiat Modares Department of Anatomy, Faculty of Medical Science, University of Tarbiat Modares Iran Gholamali Moghaddam Department of Animal Science, Faculty of Agriculture, University of Tabriz Department of Animal Science, Faculty of Agriculture, University of Tabriz Iran Younes Barzgar Department of Clinical Science, Faculty of Veterinary Medicine, University of Tabriz Department of Clinical Science, Faculty of Veterinary Medicine, University of Tabriz Iran Hamed Heidari-Vala Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran DifferentiationOsteoblastSpermatogonial stem cell 69.pdf Bruder SP, Fox BS. Tissue engineering of bone: Cell based strategies. Clin Orthop 1999;367:S68-S83.##Levenberg S, Langer R. Advances in tissue engineering. Curr Top Dev Biol 2004;61:113-134.##Mooney DJ, Mikos AG. Growing new organs. Sci Am 1999;280(4):60-65.##Carpenter MK, Rosler E, Rao MS. Characterization and differentiation of human embryonic stem cells. Cloning Stem Cells 2003;5(1):79-88.##Fenno LE, Ptaszek LM, Cowan CA. Human embryonic stem cells: emerging technologies and practical applications. Curr Opin Genet Dev 2008;18(4):324-329.##Murry CE, Keller G. Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development. Cell 2008;132 (4):661-680.##Zhang Y, Wu Y. Generation and application of pluripotent stem cells from spermatogonial stem cells. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi 2011;28(1):208-212.##Streckfuss-Bömeke K, Vlasov A, Hülsmann S, Yin D, Nayernia K, Engel W, Hasenfuss G, et al. Generation of functional neurons and glia from multipotent adult mouse germ-line stem cells. Stem Cell Res 2009;2(2):139-154. ##Huang YH, Chin CC, Ho HN, Chou CK, Shen CN, Kuo HC, et al. Pluripotency of mouse spermatogonial stem cells maintained by IGF-1- dependent pathway. FASEB J 2009;23(7):2076-2087.##Izadyar F, Spierenberg GT, Creemers LB, den Ouden K, de Rooij DG. Isolation and purification of type A spermatogonia from the bovine testis. Reproduction 2002; 124(1):85-94.##Tajik P, Barin A, Movahedin M, Zarnani AH, Hadavi R, Moghaddam G, et al. Nestin, a neuroectodermal stem cell marker, is expressed by bovine sertoli cells. Comp Clin Pathol 2010;doi:10.1007/s00580-010-1105-3. ##Kubota H, Avarbock MR, Brinster RL. Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells. Proc Natl Accad Sci USA 2004;101(47):16489-16494.##Tarfiei Gh A, Mahmoodinia Maymand M, Noruzinia M. The effect of differentiation inducing factors on osteoblastogenesis from mesenchymal stem cells. Gene 3 M 2010;7(4):1864-1870.##Liu XJ, Ren GH, Liao H, Yu L, Yuan L. Induced differentiation of adult human bone marrow derived mesenchymal stem cells in vitro toward osteoblasts. Di Yi Jun Yi Da Xue Xue Bao 2004;24(4):408-11.##Stevens LC. Spontaneous and experimentally induced testicular teratomas in mice. Cell Differ 1984;15(2-4):69-74.##Guan K, Nayernia K, Maier LS, Wagner S, Dressel R, Lee JH, et al. Pluripotency of spermatogonial stem cells from adult mouse testis. Nature 2006; 440(7088):1199-1203.##Matsui Y, Zsebo K, Hogan BL. Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture. Cell 1992;70(5):841-847.##Resnick JL, Bixler LS, Cheng L, Donovan PJ. Long-term proliferation of mouse primordial germ cells in culture. Nature 1992;359(6395):550-551.##Duplomb L, Dagouassat M, Jourdon P, Heymann D. Differentiation of osteoblasts from mouse embryonic stem cells without generation of embryoid body. In Vitro Cell Dev Biol Anim 2007;43(1):21-24.##Arpornmaeklong P, Brown SE, Wang Z, Krebsbach PH. Phenotypic characterization, osteoblastic differentiation and bone regeneration capacity of human embryonic stem cell derived mesenchymal stem cells. Stem Cells Dev 2009;18(7):955-968.##Parikka V, Vaananen A, Risteli J, Salo T, Sorsa T, Vaananen HK, et al. Human mesenchymal stem cell derived osteoblasts degrade organic bone matrix in vitro by matrix metalloproteinases. Matrix Biol 2005;24(6):438-447.##Sila-Asna M, Bunyaratvej A, Maeda S, Kitaguchi H, Bunyaratavej N. Osteoblast differentiation and bone formation gene expression in strontium-inducing bone marrow mesenchymal stem cell. Kobe J Med Sci 2007;53(1-2):25-35.##Li B, Wang XY, Tian Z, Xiao XJ, Xu Q, Wei CX, et al. Directional differentiation of chicken spermatogonial stem cells in vitro. Cytotherapy 2010;12(3):326-331.##