Editorial 2 2 As our valued readers you may well be aware that, the Avicenna Journal of Medical Biotechnology (AJMB) has recently been approved by the Commission for the Accreditation and Improvement of Iranian Medical Journals (CAIMJ) in the Ministry of Health and Medical Education. The AJMB is now included in the final list of accredited scientific and research journals in Iran and articles published in the AJMB will receive the appropriate credits from all the research/teaching institutes and Universities throughout the country. This is a great news for all our readers, contributors and particularly members of the editorial board and associates whose efforts in the past two years have brought this recognition from the Ministry of Health and Medical Education. I hope such acknowledgement will further facilitate the submission of high quality articles from the researchers in Iran who are working in the field of Medical Biotechnology. I must also inform our dear readers that the AJMB in the past year has been abstracted/ indexed in the databases as follows: Google Scholar, Genamics JournalSeek, Index Copernicus, EBSCO, Academic Search Complete, Serials Solution, Ulrichs Periodical Directory, Open J-Gate, Electronic Journals Library EZB, Scientific Information Database (SID), EMRmedex, IranMedex and Magiran. The AJMB will be added to further databases gradually as the number of published issues increases in the near future. Inclusion in the databases that index so many international journals in the field of medicine and medical biotechnology would certainly bring wider readership worldwide for the articles published in the AJMB. Therefore, I would like to invite all scientists worldwide who work in the field of Medical Biotechnology to submit their works for publication in the AJMB. We would like to consider this journal as an international journal rather than a journal of a particular geographical region. 134 134 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 154.pdf ####

Hepatic Tissue Engineering Using Scaffolds: State of the Art 2 2 Severe hepatic failure accounts for many deaths and raises medical costs each year worldwide. Currently, liver transplantation is the most common therapeutic option for patients with end-stage chronic liver disease. Due to decrease in the number of organ donors, many in need of transplantation continue to remain on the waiting list. Hepatic Tissue Engineering is a step toward alleviating the need for organ donors. Regenerative medicine and tissue engineering require two complementary key ingredients as follows: 1) biologically compatible scaffolds that can be readily adopted by the body system without harm, and 2) suitable cells including various stem cells or primary cells that effectively replace the damaged tissues without adverse consequences. Yet many challenges must be overcome such as scaffold choice, cell source and immunological barriers. Today, hepatogenic differentiation of stem cells has created trust and promise for use of these cells in hepatic tissue engineering and liver replacement. However, using suitable scaffolds is an important key to achieving the necessary functions required for hepatic replacement. In recent years, different scaffolds have been used for liver tissue engineering. In this review, we have presented different concepts in using cell /scaffold constructs to guide hepatic tissue engineering. 135 145 Somaieh Kazemnejad Department of Embryology and Stem Cells, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Department of Embryology and Stem Cells, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran HepatocyteLiverStem cellsTissue engineeringTissue scaffold 19.pdf Kuling KM, Vacanti JP. Hepatic tissue engin-eering. Transpl Immunol 2004;12(3-4):303-310. ##Lorenzini S, Andreone P. 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Sperm Chromatin Integrity: Etiologies and Mechanisms of Abnormality, Assays, Clinical Importance, Preventing and Repairing Damage 2 2 The standard semen analysis is the first line and the most popular laboratory test in the diagnosis of male fertility. It evaluates sperm concentration, motility, morphology and their vitality. However, it is well-known that normal results of semen analysis can not exclude men from the causes of couples? infertility. One of the most important parameters of sperm in its fertilizing potential is "Sperm chromatin integrity" that has direct positive correlation with Assisted Reproductive Techniques (ART) outcomes including; fertilization rate, embryo quality, pregnancy and successful delivery rate. It seems that sperm DNA chromatin integrity provides better diagnostic and prognostic approaches than standard semen parameters. For these reasons under-standing the sperm chromatin structure, etiology of sperm chromatin abnor-mality, identification factors that disturbs sperm chromatin integrity and the mechanism of their action can help in recognizing the causes of couples? infertility. Various methods of its evaluation, its importance in male fertility, clinical relevance in the outcomes of ART and application of laboratory and medical protocols to improve this integrity have valuable position in diagnosis and treatment of male infertility. There has recently been interest in the subject and its application in the field of andrology. Therefore, with regard to the above mentioned importance of sperm chromatin integrity, this review article describes details of the useful information pertaining to sperm DNA damage including the origins, assessments, etiologies, clinical aspects, and prevention of it. 147 160 Azita Hekmatdoost Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Department of Human Nutrition, Faculty of Nutrition & Food Industry, Shaheed Beheshti University of Medical Sciences Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Department of Human Nutrition, Faculty of Nutrition & Food Industry, Shaheed Beheshti University of Medical Sciences IranIran Niknam Lakpour Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mohammad Reza Sadeghi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran ChromatinDNA DamageFertilityMaleSpermatozoa 20.pdf Kidd SA, Eskenazi B, Wyrobek AJ. 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TSGA10 is Specifically Expressed in Astrocyte and Over-expressed in Brain Tumors 2 2 In this study TSGA10 has been demonstrated as a testis-specific human gene that encodes a protein localized in sperm-tail and conserved in ciliary structure. Further investigations showed TSGA10 signalling and expression during embryogenesis, brain development and some malignancies including brain tumors. Given the role of this protein in neuronal development and in certain tumors, it could potentially serve as a diagnostic marker and therapeutic target in brain tumors. Therefore, using immunohistochemistry, we evaluated the localization of TSGA10 in different regions of brain, and its pattern/level of expression in tissue microarray (Cybrdi) containing human brain tumors and normal brain. In rat specimens, TSGA10 was mainly expressed in subventricular zone, hippocampus and granular layer of cerebellum of the brain. The antibody also stained the diverse and different types of human brain cancers. The TSGA10 was strongly over-expressed in glioblastoma and astrocytoma when compared to normal human brain. The expression of TSGA10 was also confirmed in astrocyte derived from a human astroctyoma cell line by immunocytochemistry. This study indicates that TSGA10 can be used as an immunohistochemical marker for human neuroglia and astrocyte cells and is over-expressed in brain tumors. 161 166 Babak Behnam Department of Biology, University College London (UCL) Department of Microbiology and Molecular Biology, College of Medicine, University of Central Florida (UCF), Orlando Department of Genetics and Molecular Biology, College of Medicine, Iran University of Medical Sciences (IUMS) Department of Biology, University College London (UCL) Department of Microbiology and Molecular Biology, College of Medicine, University of Central Florida (UCF), Orlando Department of Genetics and Molecular Biology, College of Medicine, Iran University of Medical Sciences (IUMS) UKUSAIran Ali Chahlavi St. Vincent's Spine and Brain Institute, Jacksonville USA Jogi Pattisapu Department of Microbiology and Molecular Biology, College of Medicine, University of Central Florida (UCF), Orlando Department of Microbiology and Molecular Biology, College of Medicine, University of Central Florida (UCF), Orlando USA Jonathan Wolfe Department of Biology, University College London (UCL) Department of Biology, University College London (UCL) UK AstrocytesBrain neoplasmsImmunohistochemistryNeuron-gliaTSGA10 protein 21.pdf Behnam B, Modarressi MH, Conti V, Taylor KE, Puliti A, Wolfe J. Expression of Tsga10 sperm tail protein in embryogenesis and neural development: from cilium to cell division. Biochem Biophys Res Commun 2006;344(4):1102-1110. ##Modarressi MH, Behnam B, Cheng M, Taylor KE, Wolfe J, van der Hoorn FA. Tsga10 encodes a 65-kilodalton protein that is processed to the 27-kilodalton fibrous sheath protein. Biol Reprod 2004;70(3):608-615. ##Hagele S, Behnam B, Borter E, Wolfe J, Paasch U, Lukashev D, et al. TSGA10 prevents nuclear local-ization of the hypoxia inducible factor (HIF)-1 alpha. FEBS Lett 2006;580(15):3731-3738. ##Tanaka R, Ono T, Sato S, Nakada T, Koizumi F, Hasegawa K, et al. 2004. Over-expression of the testis-specific gene TSGA10 in cancers and its immunogenicity. Microbiol Immunol 2004;48(4): 339-345. ##Theinert SM, Pronest MM, Peris K, Sterry W, Walden P. Identification of the testis-specific pro-tein 10 (TSGA10) as serologically defined tumour associated antigen in primary cutaneous T-cell lymphoma. Br J Dermatol 2005;153(3):639-641. ##Mobasheri MB, Modarressi MH, Shabani M, Asgarian H, Sharifian RA, Vossough P, et al. Expression of the testis-specific gene, TSGA10, in Iranian patients with acute lymphoblastic leukemia (ALL). Leuk Res 2006;30(7):883-889. ##Mobasheri MB, Jahanzad I, Mohagheghi MA, Aarabi M, Farzan S, Modarressi MH. Expression of two testis-specific genes, TSGA10 and SYCP3, in different cancers regarding to their pathological features. Cancer Detect Prev 2007;31(4):296-302. ##

Antinociceptive and Antipyretic Activities of Amaranthus Viridis Linn in Different Experimental Models 2 2 Methanolic extract of whole plant of Amaranthus viridis L (MEAV), was screened for antinociceptive activity using acetic acid induced writhing test, hot plate test and tail immersion test in mice. In a similar way a screening exercise was carried out to determine the antipyretic potential of the extract using yeast induced pyrexia method in rats. Administration of the extracts was applied to both laboratory animals at the doses of 200 and 400 mg/kg body weight, respectively. The results of the statistical analysis showed that MEAV had significant (p<0.01) dose dependent antinociceptive and antipyretic properties at 200 and 400 mg/kg. Hence present investigation reveals the antinociceptive and antipyretic activities of methanolic extract of Amaranthus viridis. 167 171 Bagepalli Srinivas Ashok Kumar Department of Pharmacognosy, Sri K.V.College of Pharmacy, Chickballapur India Kuruba Lakshman Department of Pharmacognosy, PES College of Pharmacy, Bangalore India Korala Konta Narsimha Jayaveera Department of Chemistry, Jawaharlal Nehru Technological University of College of Engineering, Anantapur Department of Chemistry, Jawaharlal Nehru Technological University of College of Engineering, Anantapur India Devangam Sheshadri Shekar Department of Pharmacology, Sri K.V.College of Pharmacy, Chickballapur India Chinna SwamyVel Muragan Department of Pharmacology, Sri K.V.College of Pharmacy, Chickballapur India Bachappa Manoj Department of Pharmacognosy, Sri K.V.College of Pharmacy, Chickballapur India AnalgesicsAnimalsDose response relationshipMethanolPlant extracts 22.pdf Kirtikar KR, Basu BD. Indian Medicinal Plants. 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Effects of Sperm Chromatin Integrity on Fertilization Rate and Embryo Quality Following Intracytoplasmic Sperm Injection 2 2 Sperm chromatin integrity has been being recognized as an important factor in male fertility. During normal fertilization, high quality sperm with intact chromatin are selected through natural selection in journey from vagina to fallopian tube. However, using Assisted Reproductive Techniques, particularly ICSI, the natural selection is bypassed. Therefore sperm with DNA breakage have the opportunity to fertilize the egg which may lead to decreased embryo quality and implantation rate. The aim of this study was to evaluate the effects of sperm chromatin integrity on ICSI outcomes. A total of 200 semen samples were collected from couples undergoing ICSI and were analyzed according to WHO criteria. Each sample was evaluated for sperm chromatin integrity using four cytochemical assays and semen processing by swim up method. The ICSI was carried out according to a long-term pituitary down-regulation protocol. The correlation between sperm parameters, sperm chromatin integrity and ICSI outcomes (fertilization rate and embryo quality) was examined. The mean number of oocyte, fertilization rate and cleavage embryos per cycles was 7.5±5.0, 74.06%±25 and 5.4±3.6, respectively. There was not significant correlation between the results of chromatin assays (AO, AB, TB, and CMA3) and fertilization outcomes following ICSI. The fertilization rate was significantly higher for a group with less than 10% chromatin abnormality (p<0.05). Sperm chromatin integrity is essential for successful fertilization, embryo development and normal pregnancy. A protamine deficiency appeared to affect fertilization rate and embryo quality. However, the presence of confounding factors such as selection of spermatozoa according to normal morphology may influence the effect of sperm chromatin status on ICSI outcomes. 173 180 Mohammad Reza Sadeghi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mahshid Hodjat Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Niknam Lakpour Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Soheila Arefi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Naser Amirjannati Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Tahereh Modarresi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Hossain Hossaini Jadda Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mohammad Mehdi Akhondi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran ChromatinEmbryoFertilizationIntracytoplasmic sperm injection (ICSI)Reproductive techniquesSemen analysis 23.pdf Andersen AN, Goossens V, Ferraretti AP, Bhatta-charya S, Felberbaum R, de Mouzon J, et al. Assis-ted reproductive technology in Europe, 2004: results generated from European registers by ESHRE. Hum Reprod 2008;23(4):756-771. ##Jain T. Socioeconomic and racial disparities among infertility patients seeking care. Fertil Steril 2006;85(4):876-881. ##Palermo G, Joris H, Devroey P, Van Steirteghem AC, et al. Pregnancies after intracytoplasmic injec-tion of single spermatozoon into an oocyte. Lancet 1992;340(8810):17-18. ##Tarlatzis BC, Bili H. Survey on intracytoplasmic sperm injection: report from the ESHRE ICSI Task Force. European Society of Human Reproduction and Embryology. Hum Reprod 1998;13(Suppl 1): 165-177. ##Patient's Guide National Data Statistic. In: The pa-tient's guide to IVF clinics. Human Fertilization and Embryology Authority, London, UK, 1999. ##Karpman E, Williams DH, Lipshultz LI. IVF and ICSI in male infertility: update on outcomes, risks, and costs. Scientific WorldJournal 2005;5:922-932. ##Sills ES, Fryman JT, Perloe M, Michels KB, Tuc-ker MJ. Chromatin fluorescence characteristics and standard semen analysis parameters: correl-ations observed in andrology testing among 136 males referred for infertility evaluation. J Obstet Gynaecol 2004;24(1):74-77. ##Bianchi PG, Manicardi GC, Urner F, Campana A, Sakkas D. Chromatin packaging and morphology in ejaculated human spermatozoa: evidence of hid-den anomalies in normal spermatozoa. Mol Hum Reprod 1996;2(3):139-144. ##Boe-Hansen GB, Fedder J, Ersbøll AK, Christen-sen P. The sperm chromatin structure assay as a diagnostic tool in the human fertility clinic. Hum Reprod 2006;21(6):1576-1582. ##Evenson DP, Jost LK, Marshall D, Zinaman MJ, Clegg E, Purvis K, et al. Utility of the sperm chro-matin structure assay as a diagnostic and progno-stic tool in the human fertility clinic. Hum Reprod 1999;14(4):1039-1049. ##Silva PF, Gadella BM. Detection of damage in mammalian sperm cells. Theriogenology 2006;65 (5):958-978. ##Talbot P, Chacon RS. A triple-stain technique for evaluating normal acrosome reactions of human sperm. J Exp Zool 1981;215(2):201-208. ##Fernández JL, Muriel L, Goyanes V, Segrelles E, Gosálvez J, Enciso M, et al. Simple determination of human sperm DNA fragmentation with an im-proved sperm chromatin dispersion test. Fertil Steril 2005;84(4):833-842. ##Shoukir Y, Chardonnens D, Campana A, Sakkas D. Blastocyst development from supernumerary embryos after intracytoplasmic sperm injection: a paternal influence? Hum Reprod 1998;13(6):1632-1637. ##Larson KL, DeJonge CJ, Barnes AM, Jost LK, Evenson DP. Sperm chromatin structure assay parameters as predictors of failed pregnancy fol-lowing assisted reproductive techniques. Hum Re-prod 2000;15(8):1717-1722. ##Razavi S, Nasr-Esfahani MH, Mardani M, Mafi A, Moghdam A. Effect of human sperm chromatin anomalies on fertilization outcome post-ICSI. Andrologia 2003;35(4):238-243. ##Velez de la Calle JF, Muller A, Walschaerts M, Clavere JL, Jimenez C, Wittemer C, et al. Sperm deoxyribonucleic acid fragmentation as assessed by the sperm chromatin dispersion test in assisted reproductive technology programs: results of a large prospective multicenter study. Fertil Steril 2008;90(5):1792-1799. ##Lin MH, Kuo-Kuang Lee R, Li SH, Lu CH, Sun FJ, Hwu YM. Sperm chromatin structure assay parameters are not related to fertilization rates, embryo quality, and pregnancy rates in in vitro fertilization and intracytoplasmic sperm injection, but might be related to spontaneous abortion rates. Fertil Steril 2008;90(2):352-359. ##World Health Organization. WHO Laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 4th ed. New York: Cambridge University Press; 1999. ##Younglai EV, Holt D, Brown P, Jurisicova A, Cas-per RF. Sperm swim-up techniques and DNA frag-mentation. Hum Reprod 2001;16(9):1950-1953. ##Al-Hasani S, Demirel LC, Schöpper B, Bals-pratsch M, Nikolettos N, Küpker W, et al. Preg-nancies achieved after frozen-thawed pronuclear oocytes obtained by intracytoplasmic sperm injec-tion with spermatozoa extracted from frozen-thawed testicular tissues from non-obstructive azoospermic men. Hum Reprod 1999;14(8):2031-2035. ##Ebner T, Yaman C, Moser M, Sommergruber M, Pölz W, Tews G. Embryo fragmentation in vitro and its impact on treatment and pregnancy out-come. Fertil Steril, 2001;76(2):281-285. ##Tejada RI, Mitchell JC, Norman A, Marik JJ, Friedman S. A test for the practical evaluation of male fertility by acridine orange (AO) fluores-cence. Fertil Steril 1984;42(1):87-91. ##Hodjat M, Akhondi MA, Al-Hasani S, Mobaraki M, Sadeghi MR. Increased sperm ubiquitination correlates with abnormal chromatin integrity. Reprod Biomed Online 2008;17(3):324-30. ##Erenpreisa J, Erenpreiss J, Freivalds T, Slaidina M, Krampe R, Butikova J, et al. Toluidine blue test for sperm DNA integrity and elaboration of image cyto-metry algorithm. Cytometry A 2003;52(1):19-27. ##Singleton S, Zalensky A, Doncel GF, Morshedi M, Zalenskaya IA. Testis/sperm-specific histone 2B in the sperm of donors and subfertile patients: vari-ability and relation to chromatin packaging. Hum Reprod 2007;22(3):743-50. ##Hammadeh ME, al-Hasani S, Stieber M, Rosen-baum P, Küpker D, Diedrich K, et al. The effect of chromatin condensation (aniline blue staining) and morphology (strict criteria) of human spermatozoa on fertilization, cleavage and pregnancy rates in an intracytoplasmic sperm injection programme. Hum Reprod 1996;11(11): 2468-2471. ##Evenson D, Jost L. Sperm chromatin structure assay for fertility assessment: Current protocols in cyto-metry. Methods Cell Sci 2000;22(2-3):168-189. ##Evenson DP, Larson KL, Jost LK. Sperm chro-matin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J Androl 2002; 23(1):25-43. ##Aoki VW, Moskovtsev SI, Willis J, Liu L, Mullen JB, Carrell DT. DNA integrity is compromised in protamine-deficient human sperm. J Androl 2005; 26(6):741-748. ##Hodjat M, Akhondi MA, Amirjanati N, Savadi Shirazi E, Sadeghi MR. The comparison of four different sperm chromatin assays and their correl-ation with semen parameters. TUMJ 2008;65 (Suppl 3):33-41. ##Sculthorpe HH. Metachromasia. Med Lab Sci 1978;35(4):365-370. ##Erenpreisa J, Freivalds T, Selivanova G. Influence of chromatin condensation on the absorption spectra of nuclei stained with toluidine blue. Acta Morphol Hung 1992;40(1-4):3-10. ##Zini A, Meriano J, Kader K, Jarvi K, Laskin CA, Cadesky K. Potential adverse effect of sperm DNA damage on embryo quality after ICSI. Hum Re-prod 2005;20(12):3476-3480. ##Sakkas D, Urner F, Bianchi PG, Bizzaro D, Wag-ner I, Jaquenoud N, et al. Sperm chromatin anom-alies can influence decondensation after intracyto-plasmic sperm injection. Hum Reprod 1996;11(4): 837-843. ##Sun JG, Jurisicova A, Casper RF. Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro. Biol Reprod 1997;56(3):602-607. ##Avendaño C, Franchi A, Duran H, Oehninger S. DNA fragmentation of normal spermatozoa nega-tively impacts embryo quality and intracytoplas-mic sperm injection outcome. Fertil Steril 2009 Mar 30. [Epub ahead of print] ##Benchaib M, Lornage J, Mazoyer C, Lejeune H, Salle B, François Guerin J. Sperm deoxyribo-nucleic acid fragmentation as a prognostic indica-tor of assisted reproductive technology outcome. Fertil Steril 2007;87(1):93-100. ##Sharma RK, Said T, Agarwal A. Sperm DNA damage and its clinical relevance in assessing re-productive outcome. Asian J Androl 2004;6(2): 139-148. ##Collins JA, Barnhart KT, Schlegel PN. Do sperm DNA integrity tests predict pregnancy with in vitro fertilization? Fertil Steril 2008;89(4):823-831. ##Erenpreiss J, Spano M, Erenpreisa J, Bungum M, Giwercman A. Sperm chromatin structure and male fertility: biological and clinical aspects. Asian J Androl 2006;8(1):11-29. ##Agarwal A, Said TM. Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Update 2003;9(4):331-345. ##Oliva R. Protamines and male infertility. Hum Re-prod Update 2006;12(4):417-435. ##Cho C, Jung-Ha H, Willis WD, Goulding EH, Stein P, Xu Z, et al. Protamine 2 deficiency leads to sperm DNA damage and embryo death in mice. Biol Reprod 2003;69(1):211-217. ##Sakkas D, Urner F, Bizzaro D, Manicardi G, Bianchi PG, Shoukir Y, et al. Sperm nuclear DNA damage and altered chromatin structure: effect on fertilization and embryo development. Hum Re-prod 1998;13(Suppl 4):11-19. ##Cho C, Willis WD, Goulding EH, Jung-Ha H, Choi YC, Hecht NB, Eddy EM. Haploinsuffi-ciency of protamine-1 or -2 causes infertility in mice. Nat Genet 2001;28(1):82-86. ##Hammadeh ME, Zavos PM, Rosenbaum P, Schmidt W. Comparison between the quality and function of sperm after semen processing with two different methods. Asian J Androl 2001;3(2):125-130. ##Arny M, Quagliarello J. Semen quality before and after processing by a swim-up method: relation-ship to outcome of intrauterine insemination. Fertil Steril 1987;48(4):643-648. ##Boomsma CM, Heineman MJ, Cohlen BJ, Far-quhar C. Semen preparation techniques for intra-uterine insemination. Cochrane Database Syst Rev 2004;(3):CD004507. ##Zollner U, Zollner KP, Dietl J, Steck T. Semen sample collection in medium enhances the implantation rate following ICSI in patients with severe oligoasthenoteratozoospermia. Hum Reprod 2001;16(6):1110-1114. ##Ozmen B, Koutlaki N, Youssry M, Diedrich K, Al-Hasani S. DNA damage of human spermatozoa in assisted reproduction: origins, diagnosis, im-pacts and safety. Reprod Biomed Online 2007;14 (3):384-395. ##

The Effect of the Duration of In Vitro Maturation (IVM) on Parthenogenetic Development of Ovine Oocytes 2 2 The aim of this study was to compare the effect of time of parthenogenetic activation (22 hr versus 27 hr after In Vitro Maturation-IVM) on in vitro development of ovine oocytes using either single (Ionomycin 5 ?M for 5 min or Ethanol 7% for 7 min) or combined (ionomycin and ethanol with 6-DMAP 2 mM for 3 hr) activation treatments. The abattoir-derived in vitro matured activated oocytes were cultured in modified synthetic oviductal fluid and assessed for the cleavage, blastocyst, and hatching rates. The single-activated oocytes had a reduction in cleavage, blastocyst and hatching rates compared to the combined-activated oocytes (except for the cleavage at 27 hr). In single-treated groups the rates of cleavage and blastocyst were increased as the maturation time was extended from 22 hr to 27 hr. The numbers of total cells and Inner Cell Mass (ICM), though insignificant, were greater in combined-treated groups compared to the single treatment. The number of ICM in Eth+6-DMAP group activated at 27 hr was lower than 22 hr. Nonetheless, irrespective of the activation protocol, development to the blastocyst stage, the numbers of total cell, ICM, and cell allocation (ICM/total cells) were significantly lower in parthenogenetic than fertilized embryos. In conclusion, though the cleavage and blastocyst rates in single-treated groups were positively influenced by the extension of duration of IVM (27 hr), there was a trend of decreased numbers of total cells and ICM in slightly aged oocytes. Moreover, developmental potential of ovine parthenotes, especially in young oocytes, was improved by the addition of 6-DMAP to the activation regimen. 181 191 Abolfazl Shirazi Research Institute of Animal Embryo Technology, Shahrekord University Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Research Institute of Animal Embryo Technology, Shahrekord University Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR IranIran Amin Bahiraee Research Institute of Animal Embryo Technology, Shahrekord University Research Institute of Animal Embryo Technology, Shahrekord University Iran Ebrahim Ahmadi Research Institute of Animal Embryo Technology, Shahrekord University Research Institute of Animal Embryo Technology, Shahrekord University Iran Hassan Nazari Research Institute of Animal Embryo Technology, Shahrekord University Research Institute of Animal Embryo Technology, Shahrekord University Iran Banafsheh Heidari Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Sara Borjian Research Institute of Animal Embryo Technology, Shahrekord University Research Institute of Animal Embryo Technology, Shahrekord University Iran BlastocystEthanolIonomycinParthenogenesisSheep 24.pdf Campbell KHS. 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Biol Reprod 2000;62(1):150-154. ##Takano H, Koyama K, Kozai C, Kato Y, Tsunoda Y. Effect of aging of recipient oocytes on the development of bovine nuclear transfer embryos in vitro. Theriogenology 1993;39(4):909-917. ##Liu L, Ju J-C, Yang X. Differential inactivation of maturation-promoting factor and mitogen acti-vated protein kinase following parthenogenetic activation of bovine oocytes. Biol Reprod 1998;59 (3):537-545. ##Gordo AC, Wu H, He CL, Fissore RA. Injection of sperm cytosolic factor into mouse metaphase II oocytes induces different developmental fates ac-cording to the frequency of [Ca2+] oscillations and oocyte age. Biol Reprod 2000;62(5):1370-1379. ##Vitullo AD, Ozil JP. Repetitive calcium stimuli drive meiotic resumption and pronuclear de-velopment during mouse oocyte activation. Dev Biol 1992;151(1):128-136. ##Yazawa S, Aoyagi Y, Konishi M, Takedomi T. Characterization and cytogenetic analysis of Ja-panese Black calves produced by nuclear transfer. Theriogenology 1997;48(4):641-650. ##Liu L, Ju J-C Yang X. Parthenogenetic de-velopment and protein patterns of newly matured bovine oocytes after chemical activation. Mol Reprod Dev 1998;49(3):298-307. ##Ware CB, Barnes FL, Maiki-Laurila M, First NL. Age dependence of bovine oocyte activation. Gamete Res 1989;22(3):265-275. ##Tsunoda Y, Kato Y. Nuclear transplantation of embryonic stem cells in mice. J Reprod Fertil 1993;98:537-540. ##Taniguchi T, Taniguchi R, Kanagawa H. Influence of oocyte aging on developmental ability of re-constituted embryos produced from oocyte cyto-plast and single blastomeres of two-cell stage em-bryos. J Vet Med Sci 1996;58(7):635-640. ##Takano H, Koyama K, Kozai C, Shimizu S, Kato Y, Tsunoda Y. Effects of cell cycle stage of donor nuclei on the development of bovine nuclear transferred embryos. J Reprod Dev 1996;42(1):61-65. ##Liu G, Kato Y, Tsunoda Y. Aging of recipient oocytes reduces the development of cloned em-bryos receiving cumulus cells. 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Lack of Association Between Tumor Necrosis Factor-alpha -308 G/A Polymorphism and Risk of Developing Late-Onset Alzheimer's Disease in an Iranian Population 2 2 Late-onset Alzheimer's Disease (LOAD) is a neurodegenerative disorder and the most common form of dementia affecting people over 65 years old. Alzheimer’s disease is a complex disease with multi-factorial etiology. Inflammation has been approved to have an important role in the pathogenesis of Alzheimer’s disease (AD). TNF-a is a main pro-inflammatory cytokine that plays an essential role in initiation and regulation of inflammatory responses. Several studies have shown the probable association of polymorphism at TNF-a gene’s promoter with AD pathogenesis. This study was performed to determine whether this polymorphism contributes to the risk for late-onset Alzheimer's disease (LOAD) in Iranian population. One hundred and forty AD patients and 158 healthy controls were recruited in the study. Following extraction of genomic DNA, using PCR/RFLP methods the genotype and allele frequencies were determined in case and control subjects. The statistical analysis showed no significant difference in the allele and genotype frequencies due to this polymorphism between the two groups. Also after stratifying the subjects by their APOE-e4 status, no significant association was observed. Our results suggest that Tumor necrosis factor-alpha (TNF-?) -308 G/A is not a risk or protective factor for late-onset Alzheimer’s disease in Iranian population. However, to confirm these results further study with a bigger sample size may be required. 193 197 Mehdi Manoochehri Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Koorosh Kamali Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Epidemiology and Biostatistics Department, Tehran University of Medical Sciences Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Epidemiology and Biostatistics Department, Tehran University of Medical Sciences IranIran Mehdi Rahgozar Epidemiology and Biostatistics Department, University of Social Welfare and Rehabilitation Sciences Epidemiology and Biostatistics Department, University of Social Welfare and Rehabilitation Sciences Iran Mina Ohadi Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Homa Farrokhi Karibozorg Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Hamid Reza Khorram Khorshid Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR IranIran Alzheimer’s diseaseGenetic association studyPolymorphismPopulationTNF-aGene 25.pdf Tuppo EE, Arias HR. 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