Personalized Medicine: A Tailor Made Medicine 2 2 In the past, disease diagnosis was based on symptoms that might be indicative of several diseases. Nowadays, diagnosis of some diseases has become more accurate because we are able to test for genes known to be associated with the disease. This method not only clearly identifies the presence of a particular disease; it can also precisely determine the subtype of the disease. Throughout history, the practice of medicine has largely been reactive. Even now days, we have to wait until the onset of diseases and then try to treat or cure them. As we don’t fully understand the genetic and environmental factors that cause major diseases such as cancer, Alzheimer’s and diabetes, our efforts to treat them are often imprecise, unpredictable and ineffective. In addition, the drugs and treatments we devise are tested on broad populations and are prescribed using statistical averages. For example, on average, any given prescription drug now on the market only works for half of those who take it. Anti-depressants are effective in only about 60 percent of those who take them 1-6. Personalized medicine is beginning to transform the practice of medicine. Personalized medicine is the tailoring of medical treatment to the individual characteristics of each patient. The approach relies on scientific breakthroughs in our understanding of how a person’s unique molecular and genetic profile makes them susceptible to certain diseases. This same research is increasing our ability to predict which medical treatments will be safe and effective for each patient, and which ones will not be. Personalized medicine may be considered an extension of traditional approaches to understanding and treating disease. Personalized medicine has the potential to change the way we think about, identify and manage health problems. It is already having an exciting impact on both clinical research and patient care, and this impact will grow as our understanding and technologies improve. 191 191 Shahin Akhondzadeh Tehran University of Medical Sciences Tehran University of Medical Sciences Iran Editorial 185.pdf Akhondzadeh S, Jafari S, Raisi F, Nasehi AA, Ghoreishi A, Salehi B, et al. Clinical trial of adjunctive celecoxib treatment in patients with major depression: a double blind and placebo controlled trial. Depress Anxiety 2009;26(7):607-611.##Abolfazli R, Hosseini M, Ghanizadeh A, Ghaleiha A, Tabrizi M, Raznahan M, et al. Double-blind randomized parallel-group clinical trial of efficacy of the combination fluoxetine plus modafinil versus fluoxetine plus placebo in the treatment of major depression. Depress Anxiety 2011;28(4):297-302. ##Abbasi SH, Hosseini F, Modabbernia A, Ashrafi M, Akhondzadeh S. Effect of celecoxib add-on treatment on symptoms and serum IL-6 concentrations in patients with major depressive disorder: randomized double-blind placebo-controlled study. J Affect Disord 2012;141(2-3):308-314.##Sepanjnia K, Modabbernia A, Ashrafi M, Modabbernia MJ, Akhondzadeh S. Pioglitazone adjunctive therapy for moderate-to-severe major depressive disorder: randomized double-blind placebo-controlled trial. Neuropsychopharm-acology 2012;37(9):2093-2100.##Khajavi D, Farokhnia M, Modabbernia A, Ashrafi M, Abbasi SH, Tabrizi M, et al. Oral scopolamine augmentation in moderate to severe major depressive disorder: a randomized, double-blind, placebo-controlled study. J Clin Psychiatry 2012;73(11):1428-1433.##Shahmansouri N, Farokhnia M, Abbasi SH, Kassaian SE, Noorbala Tafti AA, Gougol A, et al. A randomized, double-blind, clinical trial comparing the efficacy and safety of Crocus sativus L. with fluoxetine for improving mild to moderate depression in post percutaneous coronary intervention patients. J Affect Disord 2014;155:216-222.##

New Insights into VEGF-A Alternative Splicing: Key Regulatory Switching in the Pathological Process 2 2 Vascular endothelial growth factor (VEGF-A) is one of the most important regulatory factors in pathological and physiological angiogenesis. Alternative splicing is a complicated molecular process in VEGF-A gene expression which adds complexity to VEGF-A biology. Among all VEGF-A exons, alternative splicing of exon 8 is the key determinant of isoform switching from pro-angiogenic VEGF-xxx to anti-angiogenic VEGF-xxxb. This is known as a key molecular switching in many pathological situations. In fact, the balance between VEGF-xxx and VEGF-xxxb isoforms is a critical controlling switch in both conditions of health and disease. Here, the properties of VEGF-xxx and VEGF-xxxb isoforms were discussed and their regulatory mechanism and their roles in certain pathological processes were evaluated. In summary, it was suggested that C-terminal VEGF-A alternative splicing can provide a new treatment opportunity in angiogenic diseases. 192 199 Fariba Dehghanian Department of Biology, Faculty of Sciences, University of Isfahan Department of Biology, Faculty of Sciences, University of Isfahan Iran Zohreh Hojati Department of Biology, Faculty of Sciences, University of Isfahan Department of Biology, Faculty of Sciences, University of Isfahan Iran Maryam Kay Department of Biology, Faculty of Sciences, University of Isfahan Department of Biology, Faculty of Sciences, University of Isfahan Iran Alternative splicingAngoigenesisDiseaseHealthVascular endothelial growth factor A 186.pdf Hoeben A, Landuyt B, Highley MS, Wildiers H, Van Oosterom AT, De Bruijn EA. Vascular endothelial growth factor and angiogenesis. Pharmacol Rev 2004;56(4):549-580.##Kajdaniuk D, Marek B, Borgiel-Marek H, Kos-Kudla B. Vascular endothelial growth factor (VEGF) part 1: in physiology and pathophysiology. 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Silky, sticky chimeras-designer VEGFs display their wares. Circ Res 2007;100(10):1402-1404. ##Dehghanian F, Hojati Z. Synthesis, cloning and mRNA expression of human VEGF-111 and VEGF-111b recombinant cDNA. (Unpublished data).##Claffey KP, Senger DR, Spiegelman BM. Structural requirements for dimerization, glycosylation, secretion, and biological function of VPF/VEGF. Biochim Biophys Acta 1995;1246(1):1-9.##Keyt BA, Berleau LT, Nguyen HV, Chen H, Heinsohn H, Vandlen R, et al. The carboxyl-terminal domain (111-165) of vascular endothelial growth factor is critical for its mitogenic potency. J Biol Chem 1996;271(13):7788-7795. ##Lee S, Jilani SM, Nikolova GV, Carpizo D, Iruela-Arispe ML. Processing of VEGF-A by matrix metalloproteinases regulates bioavailability and vascular patterning in tumors. J Cell Biol 2005;169(4):681-691. ##Lacal PM, Ruffini F, Pagani E, D'Atri S. An autocrine loop directed by the vascular endothelial growth factor promotes invasiveness of human melanoma cells. Int J Oncol 2005;27(6):1625-1632. ##Ruhrberg C, Gerhardt H, Golding M, Watson R, Ioannidou S, Fujisawa H, et al. Spatially restricted patterning cuesprovided by heparin-binding VEGF-A control blood vessel branching morphogenesis. Genes Dev 2002;16(20):2684-2698. ##Konopatskaya O, Churchill AJ, Harper SJ, Bates DO, Gardiner TA. VEGF165b, an endogenous C-terminal splice variant of VEGF, inhibits retinal neovascularisation in mice. Mol Vis 2006;12:626-632.##Seifi T, Tanhaei S, Tavassoli M, Baharvand H, Ghaedi K, Hojati Z, et al. Amplification of GC-rich putative mouse PeP promoter using betaine and DMSO in ammonium sulfate polymerase chain reaction buffer. Avicenna J Med Biotechnol 2012;4(4):206-209.##Matlin AJ, Clark F, Smith CW. Understanding alternative splicing: towards a cellular code. Nat Rev Mol Cell Biol 2005;6(5):386-398. ##Black DL. Mechanisms of alternative pre-messenger RNA splicing. Annu Rev Biochem 2003;72(1):291-336. ##Hojati Z, Dehghanian F. Enhanced production of bioactive recombinant VEGF-111 protein in Mammalian Cell Lines. Int J Biochem Cell Biol. (Unpublished data). ##Houck KA, Ferrara N, Winer J, Cachianes G, Li B, Leung DW. The vascular endothelial growth factor family: identification of a fourth molecular species and characterization of alternative splicing of RNA. Mol Endocrinol 1991;5(12):1806-1814. ##Ferrara N. Binding to the extracellular matrix and proteolytic processing: two key mechanisms regulating vascular endothelial growth factor action. Mol Biol Cell 2010;21(5):687-690. ##Krilleke D, Ng YS, Shima DT. The heparin-binding domain confers diverse functions of VEGF-A in development and disease: a structure-function study. Biochem Soc Trans 2009;37(Pt 6):1201-1206.##Bates DO, Cui TG, Doughty JM, Winkler M, Sugiono M, Shields JD, et al. VEGF165b, an inhibitory splice variant of vascular endothelial growth factor, is down-regulated in renal cell carcinoma. Cancer Res 2002;62(14):4123-4131. ##Delcombel R, Janssen L, Vassy R, Gammons M, Haddad O, Richard B, et al. New prospects in the roles of the C-terminal domains of VEGF-A and their cooperation for ligand binding, cellular signaling and vessels formation. Angiogenesis 2013;16(2):353-371.##Gu F, Li X, Kong J, Pan B, Sun M, Zheng L, et al. VEGF111b, a new member of VEGFxxxb isoforms and induced by mitomycin C, inhibits angiogenesis. Biochem Biophys Res Commun 2013;441(1):18-24.##Dehghanian F, Hojati Z. Molecular cloning and expression of VEGF-111b: A newly identified anti-angiogenic VEGF-A isoform. (Unpublished data).##Cébe Suarez S, Pieren M, Cariolato L, Arn S, Hoffmann U, Bogucki A, et al. A VEGF-A splice variant defective for heparan sulfate and neuropilin-1 binding shows attenuated signaling through VEGFR-2. Cell Mol Life Sci 2006;63(17):2067-2077. ##Dehghanian F, Hojati Z. A new approach to synthesize recombinant cDNA of VEGF-A gene by Eco31I enzyme. 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Effects of Treatment with Platinum Azidothymidine and Azidothymidine on Telomerase Activity and Bcl-2 Concentration in Hepatocellular Carcinoma- Induced Rats 2 2 Background: Telomerase activity increases in cancer cells. Bcl-2 is an antiapoptotic factor that its concentration grows in many cancer cells including hepatocellular carcinoma cells. In this study, an attempt was made to investigate the effects of a new synthetic compound, platinum azidothymidine (Pt-AZT) on treatment of rats with Hepatocellular Carcinoma (HCC) and to compare its effects with azidothymidine (AZT) in alteration of telomerase activity and Bcl-2 concentration in HCC. Methods: Healthy adult male Wistar rats (n=100) were randomly divided into 4 groups (A, B, C, and D). Group A contained 25 healthy rats and was considered as the control group. Liver preneoplastic lesions were induced in remaining animals (n=75) using Solt-Farber resistant hepatocyte protocol. These animals were randomly allocated in groups B, C and D. Group B was negative control (untreated), groups C and D were treated by intraperitoneal injection (IP) of Pt-AZT (0.9 mg/kg/day) and AZT (0.3 mg/kg/day), respectively for 14 days. After the treatment period, telomerase activity and Bcl-2 concentration were determined in the rats’ liver. Results: No HCC was developed in group A, but tumors were present in all other groups. Telomerase activity and Bcl-2 concentration were significantly lower in group C compared to groups B (0.1590.06 vs. 0.5770.116 IU/L, p<0.001, respectively and 0.9310.388 vs. 3.940.74 ng/ml, p<0.001, respectively). Similar results were observed in comparison with group D (0.3310.06 vs. 0.5770.116 IU/L, p<0.001, respectively and 0.9310.388 vs. 2.940.594 ng/ml, respectively). There was a significant negative correlation between telomerase activity and Bcl-2 concentration only in untreated cancer group (p=0.034). Conclusion: In this study, higher anticancer activity of Pt-AZT in comparison to AZT was demonstrated. It effectively inhibits the growth of liver tumor in rats through extending apoptosis. 200 209 Abdolreza Sabokrouh Department of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences Department of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences Iran Mohammad Taghi Goodarzi Research Center for Molecular Medicine, Hamadan University of Medical Sciences Research Center for Molecular Medicine, Hamadan University of Medical Sciences Iran Asad Vaisi-Raygani Molecular Diagnostic Research Center, Kermanshah University of Medical Sciences Molecular Diagnostic Research Center, Kermanshah University of Medical Sciences Iran Shohreh Khatami Department of Clinical Biochemistry, Pasteur Institute of Iran Iran Masoud Taghizadeh-Jahed Department of Tissue Engineering, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Department of Tissue Engineering, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Hepatocellular carcinomaPlatinum azidothymidineTelomerase activity 187.pdf Cong YS, Wright WE, Shay JW. 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Mitochondrial Distribution and ATP Content of Vitrified, In vitro Matured Mouse Oocytes 2 2 Background: The objective of this study was to investigate the effect of vitrification and in vitro maturation on the mitochondrial distribution and ATP content of oocytes. Methods: The oocytes at Germinal Vesicle (GV) and Metaphase II (MII) stages were recovered from 6-8 week old NMRI strain female mice. The oocytes were divided into vitrified and non-vitrified groups. Vitrification was done by the cryotop method using ethylene glycol, dimethylsulfoxide and sucrose as cryoprotectants. The GV oocytes were cultured in maturation medium for 24 hrs. The collected in vitro matured oocytes (IVM-MII) and ovulated metaphase II (OV-MII) oocytes were inseminated with capacitated sperm. The ATP content of the oocytes was measured by luciferin-luciferase reaction. Distribution of oocyte mitochondria was studied using Mito Tracker Green staining under fluorescent microscope. Results: The survival rates of vitrified oocytes at GV and MII stages were 87.39 and 89.5%, respectively. There was no significant difference in the developmental and hatching rates of vitrified and non-vitrified oocytes. The ATP content of GV and MII oocytes derived from in vivo and in vitro condition was not significantly different in vitrified and non-vitrified samples. The pattern of mitochondrial distribution in vitrified and non-vitrified GV and MII oocytes was similar but it was different between MII oocytes collected from fallopian tube and in vitro matured MII oocytes. However, the florescent intensity of mitochondrial staining was different in all the groups in the study. Conclusion: Vitrification did not affect mouse oocyte developmental competence, ATP content at different developmental stages but some alteration was seen in mitochondria distribution of in vitro matured oocytes in comparison to their controls. 210 217 Zohreh Nazmara Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Iran Mojdeh Salehnia Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Iran Saman Hosseinkhani Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University Iran ATP contentMitochondriaOocytesVitrification 188.pdf Sanchez-Partida LG, Kelly RD, Sumer H, Lo CY, Aharon R, Holland MK, et al. The generation of live offspring from vitrified oocytes. PLoS One 2011;6(6):e21597.##Rodriguez-Wallberg KA, Oktay K. Recent advances in oocyte and ovarian tissue cryopreservation and transplantation. Best Pract Res Clin Obstet Gynaecol 2012;26(3):391-405.##Wang Z, Sun Z, Chen Y, He F. A modified cryoloop vitrification protocol in the cryo preservation of mature mouse oocytes. Zygote 2009;17(3):217-224.##Campos-Chillòn LF, Suh TK, Barcelo-Fimbres M, Seidel GE Jr, Carnevale EM. Vitrification of early-stage bovine and equine embryos. Theriogenology 2009;71(2):349-354.##Turathum B, Saikhun K, Sangsuwan P, Kitiyanant Y. Effects of vitrification on nuclear maturation, ultrastructural changes and gene expression of canine oocytes. Reprod Biol Endocrinol 2010;8:70.##Vieira AD, Forell F, Feltrin C, Rodrigues JL. Calves born after direct transfer of vitrified bovine in vitro-produced blastocysts derived from vitrified immature oocytes. Reprod Domest Anim 2008;43(3):314-318.##Hurtt AE, Landim-Alvarenga F, Seidel GE, Squires EL. Vitrification of immature and mature equine and bovine oocytes in an ethylene glycol, ficoll and sucrose solution using open-pulled straws. Thriogenology 2000;54(1):119-128.##Nagai S, Mabuchi T, Hirata S, Shoda T, Kasai T, Yokota S, et al. Correlation of abnormal mitochondrial distribution in mouse oocytes with reduced developmental competence. Tohoku J Exp Med 2006;210(2):137-144.##Salehnia M, Töhönen V, Zavareh S, Inzunza J. Does cryopreservation of ovarian tissue affect the distribution and function of germinal vesicle oocytes mitochondria?. Biomed Res Int 2013:489032. ##Stojkovic M, Machado SA, Stojkovic P, Zakhartchenko V, Hutzler P, Gonçalves PB, et al. Mitochondrial distribution and adenosine triphosphate content of bovine oocytes before and after in vitro maturation: correlation with morphological criteria and developmental capacity after in vitro fertilization and culture. Biol Reprod 2001;64(3):904-909.##Liu S, Li Y, Gao X, Yan JH, Chen ZJ. Changes in the distribution of mitochondria before and after in vitro maturation of human oocytes and the effect of in vitro maturation on mitochondria distribution. Fertil Steril 2010;93(5):1550-1555.##Wang LY, Wang DH, Zou XY, Xu CM. Mitochondrial functions on oocytes and preimplantation embryos. J Zhejiang Univ Sci B 2009;10(7):483-492. ##Yu Y, Dumollard R, Rossbach A, Lai FA, Swann K. Redistribution of mitochondria leads to bursts of ATP production during spontaneous mouse oocyte maturation. J Cell Physiol 2010;224(3):672-680.##Kuwayama M, Vajta G, Leda S, Kato O. Vitrification of human embryos using the cryotip method. Reprod Biomed Online 2005;11(5):608-614.##Zavareh S, Salehnia M, Saberivand A. Comparison of different vitrification procedures on developmental competence of the mouse germinal vesicle oocytes in the presence or absence of cumulus cells. Int J Fertil Steril 2009;3(3):111-118.##Wang CY, Hitz S, Andrade JD, Stewart RJ. Specific immobilization of firefly Luciferase through a biotin carboxyl carrier protein domain. Anal Biochem 1997;246(1):133-139.##Cao Y, Xing Q, Zhang ZG, Wei ZL, Zhou P, Cong L. Cryopreservation of immature and in-vitro matured human oocytes by vitrification. Reprod Biomed Online 2009;19)3(:369-373.##Rao BS, Mahesh YU, Charan KV, Suman K, Sekhar N, Shivaji S. Effect of vitrification on meiotic maturation and expression of genes in immature goat cumulusoocyte complexes. Cryobiology 2012;64(3):176-184.##Wu G, Jia B, Mo X, Liu C, Fu X, Zhu S, et al. Nuclear maturation and embryo development of porcine oocytes vitrified by cryotop: Effect of different stages of in vitro maturation. Cryo biology 2013;67(1):95-101. ##Seet VY, Al-Samerria S, Wong J, Stanger J, Yovich JL, Almahbobi G. Optimising vitrification of human oocytes using multiple cryoprotectants and morphological and functional assessment. Reprod Fertil Dev 2013;25(6):918-926.##Habibi A, Farrokhi N, Moreira da Silva F, Bettencourt BF, Bruges-Armas J, Amidi F, et al. The effects of vitrification on gene expression in mature mouse oocytes by nested quantitative PCR. J Assist Reprod Genet 2010;27(11):599-604.##Van Blerkom J, Davis PW, Lee J. ATP content of human oocytes and developmental potential and outcome after in-vitro fertilization and embryo transfer. Hum Reprod 1995;10(2):415-424.##Brevini TA, Vassena R, Francisci C, Gandolfi F. Role of adenosine triphosphate, active mitochondria, and microtubules in the acquisition of developmental competence of parthenogenetically activated pig oocytes. Biol Reprod 2005;72(5):1218-1223.##Nayudu R, Gook D, Lopata A, Sheather S, Lloyd-Smith C, Cadusch P, et al. Follicular characteristics associated with viable pregnancy after in vitro fertilization in humans. Gamete Res 1987;18(1):37-55.##Chi MY, Manchester J, Yang V, Curato A, Strickler R, Lowry O. Contrast in levels of metabolic enzymes in human and mouse ova. Biol Reprod 1988;39(2);295-307.##Brad AM, Bormann CL, Swain JE, Durkin RE, Johnson AE, Clifford AL, et al. Glutathione and adenosine triphosphate content of in vivo and in vitro matured porcine oocytes. Mol Reprod Dev 2003;64(4):492-498.##Salvetti P, Buff S, Afanassieff M, Daniel N, Gu_erin P, Joly T. Structural, metabolic and developmental evaluation of ovulated rabbit oocytes before and after cryopreservation by vitrification and slow freezing. Theriogenology 2010;74(5):847-855.##, Carter J, DeCherney A. Effect of vitrification and thawing on human oocyte ATP concentration. Fertil Steril 2011;95(5):1839-1841.##Zhao XM, Du WH, Wang D, Hao HS, Liu Y, Qin T, et al. Recovery of mitochondrial function and endogenous antioxidant systems invitrified bovine oocytes during extended in vitro culture. Mol Reprod Dev 2011;78(12):942-950. ##Chankitisakul V, Somfai T, Inaba Y, Techakumphu M, Nagai T. Supplementation of maturation medium with L-carnitine improves cryo-tolerance of bovine in vitro matured oocytes. Theriogenology 2013;79(4):590-598.##Nishi Y, Takeshita T, Sato K, Araki T. Change of the mitochondrial distribution in mouse ooplasm during in vitro maturation. J Nippon Med Sch 2003;70(5):408-415.##

Differentiation of Human Umbilical Cord Matrix-Derived Mesenchymal Stem Cells into Germ-Like Cells 2 2 Background: Mesenchymal Stem Cells (MSCs) are multipotent cells that can be collected from different sources. Under specific conditions, MSCs can be differentiated to tissue specific cells in vitro. Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) can easily be harvested and cultured in in vitro conditions. Production of germ cells from mesenchymal stem cells is a very interesting and promising area in the field of reproductive medicine. In the present study, the possible trans-differentiation of hUCMSCs into Primordial like Germ Cell (PGC) was performed in vitro under specific condition. Methods: Human umbilical cord mesenchymal stem cells were cultured and expanded in DMEM medium containing 10% FBS. The cultured cells were studied for differentiation ability to adipocytes and osteocytes. Furthermore, MSCs related markers were identified by flow cytometry method. For PGC differentiation, hUCMS cells were cultured in differentiation medium containing Bone Morphogenetic Protein 4 (BMP4) and it was followed by retinoic acid (RA). Real time PCR and immunocytochemistry analysis were performed to evaluate the expression of PGC specific genes and proteins, respectively. Results: Our results showed that hUCMSCs cultured in the presence of BMP4 and RA are able to transdifferentiate in to PGC like cells in vitro. Real time PCR and immunocytochemistry results showed that differentiated cells expressed PGC specific markers after 14 days of culture. Conclusion: Based on these results, it was concluded that hUCMSC may be considered as a promising alternative cell source in reproductive medicine. More studies including laboratory and also animal models are needed to evaluate the functionality of differentiated PGCs before introducing them to clinical applications. 218 227 Mostafa Latifpour Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Yadollah Shakiba Department of Immunology, School of Medicine, Tehran University of Medical Sciences Department of Immunology, School of Medicine, Tehran University of Medical Sciences Iran Fardin Amidi Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Zohreh Mazaheri Department of Anatomy, School of Medicine, Tarbiat Modares University Department of Anatomy, School of Medicine, Tarbiat Modares University Iran Aligholi Sobhani Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Bone morphogenetic proteinGerm cellsMesenchymal stem cellsRetinoic acid 189.pdf International Committee for Monitoring Assisted Reproductive T, Adamson GD, de Mouzon J, Lancaster P, Nygren KG, Sullivan E, et al. World collaborative report on in vitro fertilization, 2000. Fertil Steril 2006;85(6):1586-1622.##Dimarino AM, Caplan AI, Bonfield TL. Mesenchymal stem cells in tissue repair. Front Immunol 2013;4:201.##Murphy SV, Atala A. Amniotic fluid and placental membranes: unexpected sources of highly multipotent cells. Semin Reprod Med 2013;31(1):62-68.##Batsali AK, Kastrinaki MC, Papadaki HA, Pontikoglou C. Mesenchymal stem cells derived from Wharton's Jelly of the umbilical cord: biological properties and emerging clinical applications. Curr Stem Cell Res Ther 2013;8(2):144-155.##Can A, Karahuseyinoglu S. Concise review: human umbilical cord stroma with regard to the source of fetus-derived stem cells. Stem Cells 2007;25(11):2886-2895.##Saeidi M, Masoud A, Shakiba Y, Hadjati J, Mohyeddin Bonab M, Nicknam MH, et al. Immunomodulatory effects of human umbilical cord Wharton's jelly-derived mesenchymal stem cells on differentiation, maturation and endocytosis of monocyte-derived dendritic cells. Iran J Allergy Asthma Immunol 2013;12(1):37-49.##La Rocca G, Lo Iacono M, Corsello T, Corrao S, Farina F, Anzalone R. Human Wharton's jelly mesenchymal stem cells maintain the expression of key immunomodulatory molecules when subjected to osteogenic, adipogenic and chondrogenic differentiation in vitro: new perspectives for cellular therapy. Curr Stem Cell Res Ther 2013;8(1):100-113.##Zhou C, Yang B, Tian Y, Jiao H, Zheng W, Wang J, et al. Immunomodulatory effect of human umbilical cord Wharton's jelly-derived mesenchymal stem cells on lymphocytes. Cellular Immunol 2011;272(1):33-38.##Aghaee-Afshar M, Rezazadehkermani M, Asadi A, Malekpour-Afshar R, Shahesmaeili A, Nematollahi-mahani SN. Potential of human umbilical cord matrix and rabbit bone marrow-derived mesenchymal stem cells in repair of surgically incised rabbit external anal sphincter. Dis Colon Rectum 2009;52(10):1753-1761.##Latifpour M, Nematollahi-Mahani SN, Deilamy M, Azimzadeh BS, Eftekhar-Vaghefi SH, Nabipour F, et al. Improvement in cardiac function following transplantation of human umbilical cord matrix-derived mesenchymal cells. Cardiology 2011;120(1):9-18.##Park JH, Hwang I, Hwang SH, Han H, Ha H. Human umbilical cord blood-derived mesenchymal stem cells prevent diabetic renal injury through paracrine action. Diabetes Res Clin Pract 2012;98(3):465-473.##Wang S, Yu L, Sun M, Mu S, Wang C, Wang D, et al. The therapeutic potential of umbilical cord mesenchymal stem cells in mice premature ovarian failure. BioMed Res Int 2013;2013:690491.##Qing T, Shi Y, Qin H, Ye X, Wei W, Liu H, et al. Induction of oocyte-like cells from mouse embryonic stem cells by co-culture with ovarian granulosa cells. Differentiation 2007;75(10):902-911.##Clark AT, Bodnar MS, Fox M, Rodriquez RT, Abeyta MJ, Firpo MT, et al. Spontaneous differentiation of germ cells from human embryonic stem cells in vitro. Hum Mol Genet 2004;13(7):727-739.##Nayernia K, Lee JH, Drusenheimer N, Nolte J, Wulf G, Dressel R, et al. Derivation of male germ cells from bone marrow stem cells. Lab Invest 2006;86(7):654-663.##Nayernia K, Nolte J, Michelmann HW, Lee JH, Rathsack K, Drusenheimer N, et al. In vitro-differentiated embryonic stem cells give rise to male gametes that can generate offspring mice. Dev Cell 2006;11(1):125-132.##Danner S, Kajahn J, Geismann C, Klink E, Kruse C. Derivation of oocyte-like cells from a clonal pancreatic stem cell line. Mol Hum Reprod 2007;13(1):11-20.##Hubner K, Fuhrmann G, Christenson LK, Kehler J, Reinbold R, De La Fuente R, et al. Derivation of oocytes from mouse embryonic stem cells. Science 2003;300(5623):1251-1256.##Geijsen N, Horoschak M, Kim K, Gribnau J, Eggan K, Daley GQ. Derivation of embryonic germ cells and male gametes from embryonic stem cells. Nature. 2004;427(6970):148-154.##Lacham-Kaplan O, Chy H, Trounson A. Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes. 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Quantitative Analysis of ErbB1 and ErbB2 Genes Amplification by a High Performance Liquid Chromatography 2 2 Background: Genes for human epidermal growth factor receptors B1 (ErbB1) and B2 (ErbB2) were amplified in breast and ovarian cancers. Both of them were associated with aggressive disease and worse prognosis. The ErbB1 or ErbB2 status of a tumor may provide an indication of the response to ErbB1 and ErbB2 -targeted therapies. For accurate and rapid assessment of amplification of ErbB1 and ErbB2 oncogenes, a High Performance Liquid Chromatography (HPLC) method was developed in this study. Methods: DNA was extracted from 30 primary breast tumors and 20 blood samples of healthy donors. ErbB1 and ErbB2 genes along with a reference gene were co-amplificated by Polymerase Chain Reaction (PCR). The PCR products were separated and quantified using an anion- exchange column within 30 min and in a single step. Optimum resolution was obtained when a sodium chloride gradient and a column temperature of 35˚C were used. The results of HPLC analysis of ErbB1 and ErbB2 PCR products were compared with real time PCR method as a gold standard test for 7 tumor samples. Results: The proposed HPLC method was confirmed by real time PCR method. Twenty two and ten of the specimens in our breast cancer cohort showed more than a two-fold amplification of ErbB2 and ErbB1 oncogenes, respectively. Conclusion: Our results were confirmed by real time PCR and showed that HPLC method is a specific, cheap and clinically applicable analytical approach for assessment of ErbB1 and ErbB2 statuses in breast tumors. 228 237 Mozhgan Rasti Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Iran Zohreh Honardar Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Iran Mohsen Nikseresht Department of Biochemistry, School of Medicine, Yasuj University of Medical Sciences Department of Biochemistry, School of Medicine, Yasuj University of Medical Sciences Iran Aliakbar Owji Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Iran Epidermal growth factorGene amplificationHigh pressure liquid chromatographyMethodsPolymerase chain reaction 190.pdf Bublil EM, Yarden Y. 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Outcome and human epidermal growth factor receptor (HER) 1-4 status in invasive breast carcinomas with proliferation indices evaluated by bromodeoxyuridine labelling. Breast Cancer Res 2004;6(3):R246-R251. ##Roux-Dosseto M, Romain S, Dussault N, Martin PM. Correlation of erbB-2 gene amplification with low levels of estrogen and/or progesterone receptors in primary breast cancer: do erbB-2 products delineate hormone-independent tumors? Biomed Pharmacother 1989;43(9):641-649. ##van Agthoven T, Timmermans M, Dorssers LC, Henzen-Logmans SC. Expression of estrogen, progesterone and epidermal growth factor receptors in primary and metastatic breast cancer. Int J Cancer 1995;63(6):790-793. ##Badache A, Goncalves A. The ErbB2 signaling network as a target for breast cancer therapy. J Mammary Gland Biol Neoplasia 2006;11(1):13-25.##Bouchalova K, Cizkova M, Cwiertka K, Trojanec R, Friedecky D, Hajduch M. 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Induction of Strong and Specific Humoral and T-helper 1 Cellular Responses by HBsAg Entrapped in the Methanobrevibacter smithii Archaeosomes 2 2 Background: Application of adjuvants with microbial origins is a recently highlighted approach in the vaccinology trials. Archaeosomes are among these microbial compounds with both adjuvant and liposomal activities and features. Methods: In the present study, recombinant HBsAg encapsulated into Methanobrevibacter smithii (M. smithii) archaeosomes. Balb/c mice immunized with this compound and humoral and cytokine secretion pattern of immunized models analyzed. Results: Frequency of IFN-γ secreting cells in the HBsAg-containing archaeosomes group was significantly higher than HBsAg and HBsAg+C/IFA groups (p≤0.05). IgG2a titer in the sera of HBsAg-containing archaeosomes group was also significantly higher than this subclass titer in the other groups (p≤0.05). Conclusion: Analysis of induced responses revealed the Immunopotentiating characteristics of M. smithii archaeosomes in the induction of T-helper 1 responses according to the dominance of IgG2a subtype and IFN-γ secreting splenocytes of immunized mice. 238 245 Mohammad Reza Aghasadeghi Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Seyed Ali Delbaz Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Seyed Mehdi Sadat Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Seyed Davar Siadat Department of Microbiology, Pasteur Institute of Iran Iran Mehdi Shafiee Ardestani Department of Radiopharmacy, Tehran University of Medical Sciences Department of Radiopharmacy, Tehran University of Medical Sciences Iran Pooneh Rahimi Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Azam Bolhassani Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Rouhollah Vahabpour Roudsari Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Golnaz Bahramali Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Fateme Motevalli Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Mehdi Davari Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Habib Vakily Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran Ali Sharifat Salmani Department of Biology, Science and Research Branch, Islamic Azad University Department of Biology, Science and Research Branch, Islamic Azad University Iran Maryam Borhan Nobari Department of Hepatitis and AIDS, Pasteur Institute of Iran Iran CellularHepatitis B surface antigensHumoralImmunityMethanobrevibacter 191.pdf Rosenthal KS, Zimmerman DH. Vaccines: All things considered. Clin Vaccine Immunol 2006;13(8):821-829.##Shen E, Li L, Li L, Feng L, Lu L, Yao Z, et al. PIKA as an adjuvant enhances specific humoral and cellular immune responses following the vaccination of mice with HBsAg plus PIKA. Cell Mol Immunol 2007;4(2):113-120.##Krishnan L, Dicaire CJ, Patel GB, Sprott GD. Archaeosome vaccine adjuvants induce strong humoral, cell-mediated, and memory responses: Comparison to conventional liposomes and alum. Infect Immun 2000;68(1):54-63.##Aghasadeghi MR, Salmani AS, Sadat SM, Javadi F, Memarnejadian A, Vahabpour R, et al. Application of outer membrane vesicle of Neisseria meningitidis serogroup B as a new adjuvant to induce strongly Th1-oriented responses against HIV-1. Curr HIV Res 2011;9(8):630-635. ##Sprott GD, Patel GB, Krishnan L. Archaeobacterial ether lipid liposomes as vaccine adjuvants. Methods Enzymol 2003;373:155-172.##Sprott, GD, Brisson JR, Dicaire CJ, Pelletier AK, Deschatelets AJ, Krishnan L, et al. 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Variable Number Tandem Repeat (VNTR) Genotyping of Hydatidiform Mole in Iranian Patients 2 2 Background: Classification of molar gestation into Complete Hydatidiform Mole (CHM) and Partial Hydatidiform Mole (PHM) is done according to clinical, ultrasonographic, histologic and genetic criteria. However, making a distinction between CHM and PHM using histologic criteria alone may be difficult and several studies have shown that misclassifications are frequent, even for experienced pathologists. CHM is the most common precursor to choriocarcinoma and heterozygous moles carry an increased predisposition to transformation. Methods: Formalin-fixed, paraffin-embedded tissue sections of patients as well as peripheral blood of patients and their partners’ were collected in EDTA tubes. Tissue samples were obtained by curettage. Histological evaluation was performed on routine section stained with Hematoxylin and Eosin. Variable Number Tandem Repeats (VNTRs) genotyping was performed for 30 cases in two groups of CHM (n=21) and PHM (n=9), with Polymerase Chain Reaction (PCR) amplification of 2 different polymorphic loci, namely the Col2A1 and D1S80. Results: The results of DNA analysis by VNTR genotyping showed that in 16 cases of CHM, amplification of the VNTR polymorphic loci showed androgenetic mono-spermic moles (homozygote) and in 5 cases of CHM androgenetic dispermic moles (heterozygote) in molar tissue. In cases of PHM, 6 samples were triploid dispermic and 3 samples were diploid biparental. Conclusion: This study confirmed that VNTR genotyping can identify the parental source of polymorphic alleles in hydatidiform mole. Compared to STR genotyping, VNTR genotyping was performed by PCR amplification of several minisatellite markers of DNA. This method significantly requires less time and is cost-effective. 246 253 Zahra Pakzad Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Iran Hossein Mozdarani Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Iran Narges Izadi-Mood Mirza Koochak Khan Hospital, Iran University of Medical Sciences Mirza Koochak Khan Hospital, Iran University of Medical Sciences Iran Shirin Niromanesh Mirza Koochak Khan Hospital, Iran University of Medical Sciences Mirza Koochak Khan Hospital, Iran University of Medical Sciences Iran Hydatidiform moleMinisatelliteVariable Number Tandem RepeatsGenotyping techniques 192.pdf Landolsia H, Missaouia N, Brahemc S, Hmissaa S, Gribaac M, Yacoubia MT. 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Int J Gynaecol Obstet 1978;15(5):390-395.##Slim R, Coullin Ph, Diatta AL,Chebaro W, Courtin D, Abdelhak S, and Garcia A. NLRP7 and the genetics of post-molar choriocarcinomas in Senegal. Mol Hum Reprod 2012;18(1):52-56.##Slim R, Wallace EP. NLRP7 and the genetics of hydatidiform moles: recent advances and new challenges. Front Immunol 2013;4:1-12.##Baasanjav B, Usui H, Kihara M, Kaku H, Nakada E, Tate S, et al. The risk of post-molar gestational trophoblastic neoplasia is higher in heterozygous than in homozygous complete hydatidiform moles. Hum Reprod 2010;25(5):1183-1191. ##Lai CY, Chan KY, Khoo US, Ngan HY, Xue WC, Chiu PM, et al. Analysis of gestational trophoblastic disease by genotyping and chromosome in situ hybridization. Mod Pathol 2004;17(1):40-48.##Devriendt K. Hydatidiform mole and triploidy: the role of genomic imprinting in placental development. Human Reprod Update 2005;11: 137-142. ##Fukunaga M. 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