DNA Methylation and Its Role in the Development of Leukemia 2 2 Epigenetic changes play an essential role in cancer pathogenesis. It has been established by next-generation sequencing that more than 50% of the human cancers carry mutations in mechanisms involved in the organization of the chromatin and epigenetic regulations. DNA methylation is among the most common epigenetic changes in leukemia. In contrast to DNA mutations which are passively inherited from DNA replication, epimutations, for example, the hypermethylation and epigenetic silencing of tumor suppressor genes, must be actively maintained because of being reversible. Actually, the reversibility of epimutations by small-molecule inhibitors provides the basis for the use of such inhibitors in new cancer therapy strategies. However, DNA methylation mechanism and its role in leukemia are not fully understood; there are some serious concerns about the use of these drugs. In this study, we will review the mechanisms of DNA methylation and the genes that are methylated in leukemia. Moreover, new interesting findings of the epigenetic changes causeed by adult T-cell leukemia/lymphoma have been fully discussed. 153 165 2019/02/9 1397/11/20 2019/07/20 1398/4/29 پریسا باقری Parisa Bagheri Department of Hematology and Blood banking, School of Allied Medical Sciences, Mashhad university of Medical Sciences, Mashhad, Iran ایران Bagheri.parisa2020@yahoo.com شاهین آغه میری Shahin Aghamiri Student research committee, Department of medical biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران shahin.aghamiri@gmail.com علی جعفرپور ali Jafarpour Students’ Scientific Research Center, Virology Division, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran ایران aligafarpour10101370@yahoo.com محمد علی اسماعیلی Mohammad Ali Esmaili Department of Hematology, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran ایران esmailionline@gmail.com محمد حدادی اقدم Mohammad Haddadi Aghdam Molecular Medicine Department, Biotechnology Research Center, Pasteur Institute of Iran ایران Mohammad858786@gmail.com فرهاد ذاکر Farhad Zaker 6Department of Hematology and Blood Banking, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran. ایران سوده نامجو Soodeh Namjoo Department of Hematology and Blood Banking, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran. ایران nsoodeh@yahoo.com Epigenetic Leukemia Methylation Neoplasm Treatment [1]. 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Anti-Trichomonas Vaginalis Activity of Ethanolic Extracts of Medicago Sativa and Satureja Hortensis, In Vitro Study 2 2 Background and Aims: Trichomonas vaginalis is a flagellated protozoa that is associated with vaginitis, cervicitis, urethritis and other vaginal disorders. Current study aimed to evaluate the anti-Trichomonas activity of Medicago sativa and Satureja hortensis, in vitro. Materials and Methods: Ethanolic extract of Medicago sativa and Satureja hortensis were obtained by rotary evaporator. anti-Trichomonas vaginalis activities of the extracts in different concentrations were evaluated after 24, 48 and 72 hr of incubation of the cultured media. Results and Conclusions: The data showed a significant difference between concentration and time regarding the Satureja hortensis and Medicago sativa extracts compared to the negative control (p<0.05). According to the results, the anti- trichomonas activity of the Medicago sativa and Satureja hortensis extracts may make it possible to use them in the treatment of trichomoniasis. 166 171 2019/02/92018/04/16 1397/1/27 2019/07/202019/07/15 1398/4/24 فرزانه میرزایی Farzaneh Mirzaei Department of Parasitology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران mirzaei.farzaneh@yahoo.com وحید رئیسی Vahid Raissi Department of Medical Parasitology and Mycology, School of Public Health, Tehran university of Medical Sciences, Tehran, Iran ایران vahidraissi66@gmail.com عارف تیموری Aref Teimouri Department of Medical Parasitology and Mycology, School of Public Health, Tehran university of Medical Sciences, Tehran, Iran ایران پریسا موسوی Parisa Mousavi Department of Medical Parasitology and Mycology, School of Public Health, Tehran university of Medical Sciences, Tehran, Iran ایران محمدعلی محقق Mohammad Ali Mohaghegh Department of Laboratory Sciences, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran. ایران mohaghegh1982@yahoo.com مهدی دهقان منشادی Mahdi Dehghan-manshadi Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران فاطمه زارع Fateme zare Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران بهمن رحیمی اسبوی Bahman Rahimi Esboei Department of Medical Parasitology and Mycology, School of Public Health, Tehran university of Medical Sciences, Tehran, Iran. ایران bahman51640@yahoo.com Medicago sativa Satureja hortensis Trichomonas vaginalis Satureja hortensis Medicago sativa Trichomonas vaginalis invitro [1]. Frasson AP, dos Santos O, Duarte M, da Silva Trentin D, Giordani RB, da Silva AG, et al. First report of anti-Trichomonas vaginalis activity of the medicinal plant Polygala decumbens from the Brazilian semi-arid region, Caatinga. Parasitol Res. 2012; 110(6): 2581-587.##[2]. World Health Organization. Guidelines for predicting dietary intake of pesticide residues. 1997.##[3]. Tagboto S, Townson S. Antiparasitic properties of medicinal plants and other naturally occurring products. Adv Parasitol. 2001; 50: 199-295.##[4]. El Kassas HY, Attia AA. Bactericidal applic-ation and cytotoxic activity of biosynthesized silver nanoparticles with an extract of the red seaweed Pterocladiella capillacea on the HepG2 cell line. Asian Pac J Cancer Prev. 2014; 15(3): 1299-306.##[5]. DD’addabbo T, Carbonara T, Leonetti P, Radicci V, Tava A, Avato P. Control of plant parasitic nematodes with active saponins and biomass from Medicago sativa. Phytochemistry Reviews. 2011; 10(4): 503-19.##[6]. Upcroft JA, Dunn LA, Wright JM, Benakli K, Upcroft P, Vanelle P. 5-Nitroimidazole drugs effective against metronidazole-resistant Tricho-monas vaginalis and Giardia duodenalis. Antimicrobial agents and chemotherapy 2006; 50(1): 344-47.##[7]. Mirzaei F, Bafghi AF, Mohaghegh MA, Jaliani HZ, Faridnia R, Kalani H. In vitro anti-leishmanial activity of Satureja hortensis and Artemisia dracunculus extracts on Leishmania major promastigotes. J Parasitic Dis. 2016; 40(4): 1571-574.##[8]. Upcroft P, Upcroft J. Drug targets and mechanisms of resistance in the anaerobic protozoa. Clin. Microbiol. Rev. 2001; 14(1): 150.##[9]. Brandelli CL, Vieira PD, Macedo AJ, Tasca T. Remarkable anti-trichomonas vaginalis activity of plants traditionally used by the Mbyá-Guarani indigenous group in Brazil. BioMed Res Int. 2013(2013): 1-7.##[10]. El-Sherbini GT, El BG, Abdel-Hady NM, Morsy TA. Efficacy of two plant extracts against vaginal trichomoniasis. J Egypt Society Parasitol. 2009; 39(1): 47-58.##[11]. Calzada F, Yépez-Mulia L, Tapia-Contreras A. Effect of Mexican medicinal plant used to treat trichomoniasis on Trichomonas vaginalis trophozoites. J Ethnopharmacol. 2007; 113(2): 248-51.##[12]. . Diba K, Ghabaie K, Heshmatian B, Sharbatkhori M. Antifungal activity of Satureja hortensis alcoholic extract against Aspergillus and Candida species. J. Med. Plants Res. 2013; 7(30): 2271-274.#### ##

The Effect of Follicular Fluid on the Proliferation and Osteoblastic Differentiation of Human Bone Marrow Mesenchymal Stem Cells 2 2 Background and Aims: Bone marrow-derived mesenchymal stem cells (BM-MSCs) are a well-known source of multipotent adult stem cells. Despite using different methodologies of MSCs preparing for clinical applications, the top safest procedure to manipulate these cells, has not yet been determined. Recently, ex-vivo expansion of MSCs for their subsequent implantation, using some biological product, is suggested instead of fetal bovine serum (FBS). Previous studies have shown the effect of follicular fluid (FF) (a dynamic fluid in ovarian follicle) as an additive component in cell culture. Hence, this study aimed to decipher its role on the human BM-MSC proliferation. Materials and Methods: In this study, BM-MSCs at 3rd passage were cultivated in the presence of 20% FF (group I), 10% FF+ FBS 10% (group II) and FBS 20% as control group. The capacity of proliferation as calculating population doubling times and gene expression levels of stem cell factor, stromal cell-derived factor 1, and transforming growth factor beta were analyzed in osteogeneic media to examine the impacts of FF on osteogenesis of MSCs. Results: Our results corroborated an up-regulatory effect of FF on the proliferation of BM-MSCs by shorter population doubling times in the group II of treated cells and an increase in gene expression level of osteocalcin and transforming growth factor beta in the presence of higher concentrations of FF in cell culture FF 20% and 10%, respectively. Conclusions: FF is a potent mitogen for cell proliferation. FF may be an efficient substitution of FBS in ex-vivo cell culture, eliminating zoonotic infections and immunological reactions. 172 183 2019/02/92018/04/162019/03/13 1397/12/22 2019/07/202019/07/152019/07/20 1398/4/29 عاطفه سلطانی Atefeh Soltani Department of Hematology and Blood Banking, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران atefeh.soltani@modares.ac.ir سعید آبرون Saeid Abroun Department of Hematology and Blood Banking, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران abroun@modares.ac.ir مجتبی رضازاده ولوجردی Mojtaba Rezazadeh Valojerdi Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. Department of Embryology, Royan Institute, Tehran, Iran. ایران mr_valojerdi@modares.ac.ir بهاره وحیدیان فر Bahareh Vahidianfar Department of Hematology and Blood Banking, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران Bahareh.vahidianfar@modares.ac.ir الهه حسینی Elahe Sadat Hosseini Department of Hematology and Blood Banking, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران elahehosseini@modares.ac.ir Differentiation Follicular fluid Mesenchymal stem cells Osteogenesis [1]. Payushina O, Domaratskaya E, Starostin V. Mesenchymal stem cells: sources, phenotype, and differentiation potential. Biology Bulletin 2006; 33(1): 2-18.##[2]. Baksh D, Song L, Tuan R. Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med. 2004; 8(3): 301-16.##[3]. Porcellini A. Regenerative medicine: a review. Revista Brasileira de Hematologia e Hemo-terapia 2009; 31(2): 63-6.##[4]. Kandoi S, Patra B, Vidyasekar P, Sivanesan D, Vijayalakshmi S, Rajagopal K, et al. Evaluation of platelet lysate as a substitute for FBS in explant and enzymatic isolation methods of human umbilical cord MSCs. Scientific Rep. 2018; 8(1): 12439.##[5]. Rohban R, Pieber TR. Mesenchymal stem and progenitor cells in regeneration: tissue specificity and regenerative potential. Stem Cells Int. 2017; 2017: 5173732.##[6]. Gentile P, Garcovich S. Advances in regenerative stem cell therapy in androgenic alopecia and hair loss: Wnt pathway, growth-factor, and mesenchymal stem cell signaling impact analysis on cell growth and hair follicle development. Cells 2019; 8(5): 466.##[7]. Dessels C, Potgieter M, Pepper MS. Making the switch: alternatives to fetal bovine serum for adipose-derived stromal cell expansion. Front Cell Dev Biol. 2016; 4(5): 115.##[8]. Revelli A, Delle Piane L, Casano S, Molinari E, Massobrio M, Rinaudo P. Follicular fluid content and oocyte quality: from single biochemical markers to metabolomics. Reproduct Biol Endocrinol. 2009; 7(1): 40.##[9]. Ambekar AS, Nirujogi RS, Srikanth SM, Chavan S, Kelkar DS, Hinduja I, et al. Proteomic analysis of human follicular fluid: a new perspective towards understanding folliculo-genesis. J Proteomic. 2013; 87(1): 68-77.##[10]. Rodgers RJ, Irving-Rodgers HF. Formation of the ovarian follicular antrum and follicular fluid. Biol Reproduc. 2010; 82(6): 1021-1029.##[11]. Qiu P, Bai Y, Liu C, He X, Cao H, Li M, et al. A dose-dependent function of follicular fluid on the proliferation and differentiation of umbilical cord mesenchymal stem cells (MSCs) of goat. Histochem Cell Biol. 2012; 138(4): 593-603.##[12]. Bedaiwy M, Shahin AY, AbulHassan AM, Goldberg JM, Sharma RK, Agarwal A, et al. Differential expression of follicular fluid cytokines: relationship to subsequent pregnancy in IVF cycles. Reproduct Biomed Onlin. 2007; 15(3): 321-25.##[13]. Chen FP, Hu CH, Wang KC. Estrogen modulates osteogenic activity and estrogen receptor mRNA in mesenchymal stem cells of women. Climacteric 2013; 16(1): 154-60.##[14]. Qiu X, Jin X, Shao Z, Zhao X. 17β-estradiol induces the proliferation of hematopoietic stem cells by promoting the osteogenic differentiation of mesenchymal stem cells. Tohoku J Exper Med. 2014; 233(2): 141-48.##[15]. Leskela HV, Olkku A, Lehtonen S, Mahonen A, Koivunen J, Turpeinen M, et al. Estrogen receptor alpha genotype confers interindividual variability of response to estrogen and testosterone in mesenchymal-stem-cell-derived osteoblasts. Bone 2006; 39(5): 1026-1034.##[16]. Tsao YT, Huang YJ, Wu HH, Liu YA, Liu YS, Lee OK. Osteocalcin mediates biomineralization during osteogenic maturation in human mesenchymal stromal cells. Int J Mol Sci. 2017; 18(1): 1-10.##[17]. Figueira MI, Cardoso HJ, Correia S, Maia CJ, Socorro S. Hormonal regulation of c-KIT receptor and its ligand: implications for human infertility? Progress in histochemistry and cytochemistry. 2014; 49(1-3): 1-19.##[18]. Li X, Jin L, Cui Q, Wang GJ, Balian G. Steroid effects on osteogenesis through mesenchymal cell gene expression. Osteoporos Int. 2005; 16(1): 101-108.##[19]. Chabanon A, Desterke C, Rodenburger E, Clay D, Guerton B, Boutin L, et al. A cross-talk between stromal cell-derived factor-1 and transforming growth factor-beta controls the quiescence/cycling switch of CD34(+) progenitors through FoxO3 and mammalian target of rapamycin. Stem Cells (Dayton, Ohio). 2008; 26(12): 3150-161.##[20]. Knight PG, Glister C. Local roles of TGF-beta superfamily members in the control of ovarian follicle development. Animal Reproduct Sci. 2003; 78(3-4): 165-83.##[21]. Virant-Klun I, Skutella T, Kubista M, Vogler A, Sinkovec J, Meden-Vrtovec H. Expression of pluripotency and oocyte-related genes in single putative stem cells from human adult ovarian surface epithelium cultured in vitro in the presence of follicular fluid. Biomed Res Int. 2013; 2013: 861460.##[22]. Zajdel A, Kalucka M, Kokoszka-Mikolaj E, Wilczok A. Osteogenic differentiation of human mesenchymal stem cells from adipose tissue and Wharton's jelly of the umbilical cord. Acta biochimica Polonica. 2017;64(2): 365-69.##[23]. da Silva Meirelles L, Chagastelles PC, Nardi NB. Mesenchymal stem cells reside in virtually all post-natal organs and tissues. J Cell Sci. 2006; 119(11): 2204-213.##[24]. Barry FP. Biology and clinical applications of mesenchymal stem cells. Birth Defects Res: Embryo Today 2003; 69(3): 250-56.##[25]. Lai D, Guo Y, Zhang Q, Chen Y, Xiang C. Differentiation of human menstrual blood-derived endometrial mesenchymal stem cells into oocyte-like cells. Acta Biochimica Biophysica Sinica. 2016; 48(11): 998-1005.##[26]. Yoo SW, Savchev S, Sergott L, Rezai T, Lopez MF, Von Wald T, et al. A large network of interconnected signaling pathways in human ovarian follicles is supported by the gene expression activity of the granulosa cells. Reprod Sci. 2011; 18(5): 476-84.##[27]. Mirzaeian L, Eftekhari-Yazdi P, Esfandiari F, Eivazkhani F, Rezazadeh Valojerdi M, Moini A, et al. Induction of mouse peritoneum mesenchymal stem cells into germ cell-like cells using follicular fluid and cumulus cells conditioned media. Stem Cells Dev. 2019; 28(8): 554-64.##[28]. Basuino L, Silveira CF. Human follicular fluid and effects on reproduction. JBRA Assist Reproduct. 2016; 20(1): 38-40.##[29]. Hsieh M, Zamah AM, Conti M. Epidermal growth factor-like growth factors in the follicular fluid: role in oocyte development and maturation. Seminars Reproduct Med. 2009; 27(1): 52-61.##[30]. Volarevic V, Gazdic M, Simovic Markovic B, Jovicic N, Djonov V, Arsenijevic N. Mesenchymal stem cell-derived factors: Immuno-modulatory effects and therapeutic potential. BioFactors 2017; 43(5): 633-44.##[31]. Lennartsson J, Ronnstrand L. Stem cell factor receptor/c-Kit: from basic science to clinical implications. Physiol Rev. 2012; 92(4): 1619-649.## ##

Genotyping of Mycobacterium Tuberculosis Isolated from Suspected Patients in Tehran in 2015-2017 2 2 Background and Aims: Unlike many global efforts to eradicate tuberculosis caused by Mycobacterium, it remains as a life-threatening infection with a worldwide incidence of 1.5 million cases each year. However, due to the lack of information about Mycobacterium tuberculosis characterization, more studies are required to evaluate strain diversity and epidemiology of tuberculosis to improve the therapeutic approaches. This study aimed to genotype the Mycobacterium tuberculosis isolated from suspected patients in Tehran, Iran through 2015-2017. Materials and Methods: In the current study, 30 isolates (sputum, broncho-alveolar lavage and biopsy) were collected from different tuberculosis patients at Massoud Clinical Lab of Tehran from 2015 to 2017. To find the single nucleotide polymorphisms and mutated regions, polymerase chain reaction (PCR) was performed on all the isolates to amplify the katG and gyrA genes. Then, PCR products were sequenced and analyzed. Results: The majority of isolates were assigned to PGG2 (90%), followed by PGG3 (10%) but no isolate belonging to PGG1 was found. Conclusions: Our findings demonstrate a remarkable epidemiological pattern of tuberculosis in Tehran. In group 2, isolates showed a considerably higher frequency compared to isolates in group 3, which is consistent with other findings reported in Iran. However, in contrast to other Iranian studies, no isolated strains were categorized in principal PGG1. 184 191 2019/02/92018/04/162019/03/132019/01/9 1397/10/19 2019/07/202019/07/152019/07/202019/07/2 1398/4/11 سعید ذاکر Saeed Zaker Bostanabad Department of Biology and Biotechnology, Islamic Azad University, Parand branch, Iran. ایران saeedzaker20@yahoo.com هانیه باقریفرد Hanieh Bagherifard Department of Microbiology, Islamic Azad University, Damghan branch, Iran. ایران saeedzaker20@yahoo.com Genotyping gyrA katG Mycobacterium tuberculosis [1]. Frieden TR, Sterling TR, Munsiff SS, Watt CJ, Dye C. Tuberculosis. Lancet 2003; 362(9387): 887-99.##[2]. Lari N, Rindi L, Sola C, Bonanni D, Rastogi N, Tortoli E, et al. Genetic diversity, determined on the basis of katG463 and gyrA95 polymorphisms, Spoligotyping, and IS6110 typing, of Mycobacterium tuberculosis complex isolates from Italy. J clin microbiol. 2005; 43(4): 1617-624.##[3] . World Health Organization. Global health observatory data repository 2013. Available at: who int/gho/data/node main A. 2013; 364.##[4] . Sreevatsan S, Pan X, Stockbauer KE, Connell ND, Kreiswirth BN, Whittam TS, et al. Restricted structural gene polymorphism in the Myco-bacterium tuberculosis complex indicates evolutionarily recent global dissemination. Proc Natl Acad Sci USA 1997; 94(18): 9869-874.##[5]. Kim S, Misra A. SNP genotyping: technologies and biomedical applications. Ann Rev Biomed Eng. 2007; 9(1): 289-320.##[6] . Royo JL, Galan JJ. Pyrosequencing for SNP genotyping. Methods in molecular biology 2009; 578: 123-33. ##[7]. Li R, Li Y, Fang X, Yang H, Wang J, Kristiansen K, et al. SNP detection for massively parallel whole-genome resequencing. Genome Res. 2009; 19(6): 1124-132. ##[8] . Ravansalar H, Tadayon K, Ghazvini K. Molecular typing methods used in studies of Mycobacterium tuberculosis in Iran: a systematic review. Iran J Med Microbiol. 2016; 8(5): 338-46.##[9]. McEvoy CR, van Pittius NCG, Victor TC, van Helden PD, Warren RM. The role of IS6110 in the evolution of Mycobacterium tuberculosis. Tuberculosis 2007; 87(5): 393-404.##[10]. Roychowdhury T, Mandal S, Bhattacharya A. Analysis of IS6110 insertion sites provide a glimpse into genome evolution of Mycobacterium tuberculosis. Sci Rep. 2015; 5(1): 12567.##[11] . Poulet S, Cole ST. Repeated DNA sequences in mycobacteria. Arch Microbiol. 1995; 163(2): 79-86.##[12] . Poulet S, Cole ST. Characterization of the highly abundant polymorphic GC-rich-repetitive sequence (PGRS) present in Mycobacterium tuberculosis. Arch Microbiol. 1995; 163(2): 87-95.##[13] . Stucki D, Malla B, Hostettler S, Huna T, Feldmann J, Yeboah-Manu D, et al. Two new rapid SNP-typing methods for classifying Mycobacterium tuberculosis complex into the main phylogenetic lineages. PloS One 2012; 7(7): 41253.##[14]. Ford CB, Lin PL, Chase MR, Shah RR, Iartchouk O, Galagan J, et al. Use of whole genome sequencing to estimate the mutation rate of Mycobacterium tuberculosis during latent infection. Nat Genet. 2011; 43(5): 482-86.##[15] . Petroff S. A new and rapid method for the isolation and cultivation of tubercle bacilli directly from the sputum and feces. J Exp Med. 1915; 21(1): 38-42.##[16]. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. 2nd ed, NY: Cold spring harbor laboratory Cold Spring Harbor; 1989.##[17]. Bouakaze C, Keyser C, De Martino S, Sougakoff W, Veziris N, Dabernat H, et al. Identification and genotyping of Mycobacterium tuberculosis complex species by use of a SNaPshot Minisequencing-based assay. J Clin Microbiol 2010; 48(5): 1758-766.##[18] . The NCBI handbook. National Library of Medicine (US), N.C.f.B.I.O.C., The Reference Sequence (RefSeq) Project [Internet]. Available at:http://www.ncbi.nlm.nih.gov/nuccore/448814763/?report=genbank.##[19] . Dou HY, Lin CH, Chen YY, Yang SJ, Chang JR, Wu KM, et al. Lineage-specific SNPs for genotyping of Mycobacterium tuberculosis clinical isolates. Sci Rep. 2017; 7(1): 1425.##[20]. Coll F, McNerney R, Guerra-Assuncao JA, Glynn JR, Perdigao J, Viveiros M, et al. A robust SNP barcode for typing Mycobacterium tuberculosis complex strains. Nat commun. 2014; 5(2): 4812.##[21]. de Viedma DG, Mokrousov I, Rastogi N. Innovations in the molecular epidemiology of tuberculosis. Enferm Infecc Microbiol Clin. 2011; 29(1): 8-13. ##[22] . Eshghinejad A, Farazi A, Eshrati B, Khalili H, Shojapour M, Ahmadi A, et al. Detection of major genetic groups of mycobacterium tuberculosis isolated from tuberculosis patients in Markazi province by polymorphism determination in kat G and gyr A genes. J Arak Uni Med Sci. 2012; 15 (5): 26-34.##[23] . Zaker Bostanabad S, Titov LP, Slizen VV, Taghikhani M, Bahrmand A. katG mutations in isoniazid-resistant strains of Mycobacterium tuberculosis isolates from Belarusian patients. Tuberkuloz ve toraks. 2007; 55(3): 231-37.## ##

Molecular Detection of Adenoviruses in the Sinus Tissues of Patient by Nested-PCR in Shiraz, Southwest Iran 2 2 Background and Aims: Rhinosinusitis is an inflammation of the mucous membrane that may be caused by infectious agents such as bacteria, fungi and viruses. Few studies have been carried out on the role of viruses in Rhinosinusitis patients .The aim of this study was the molecular detection of Adenoviruses in sinus tissues by nested polymerase chain reaction (PCR) in Shiraz. Materials and Methods: In the present study, 103 paraffin-embedded biopsy specimens of sinus tissues were subjected to DNA extraction and tested for adenovirus DNA using Nested PCR. The amplification of a β-globin gene by PCR-based method was used to confirm the quality of extracted DNA. Results: A total of 103 samples of sinus tissues were examined. Of these patients, 50 (48.54%) were male and the rest were female (51.46%). The patients’ age ranged between 2 and 82 years and the mean age was 42.15±1.56 years. The adenovirus DNA was detected in 13 of 103 (12.6%) samples. Conclusions: The results of this study showed that Adenoviruses have high prevalence in rhinosinusitis patients. As a results, it is an important to investigate clinical significance of viral infections especially Adeno viruses in these patients. 192 198 2019/02/92018/04/162019/03/132019/01/92018/09/25 1397/7/3 2019/07/202019/07/152019/07/202019/07/22019/07/2 1398/4/11 شهاب محمد وند Shahab Mahmoudvand Department of Bacteriology & Virology, Faculty of Medicine, Shiraz University of Medical Sciences Shiraz, Iran. ایران mahmoudvandsh100@yahoo.com محمد کرد Mohammad Kord Department of Mycology, Faculty of Medicine, Shiraz University of Medical Sciences Shiraz, Iran. ایران mahmoudvandsh100@yahoo.com ندا پیربنیه Neda Pirbonyeh Department of Bacteriology & Virology, Faculty of Medicine, Shiraz University of Medical Sciences Shiraz, Iran. ایران mahmoudvandsh100@yahoo.com افاق معطری Afagh Moattari Department of Bacteriology & Virology, Faculty of Medicine, Shiraz University of Medical Sciences Shiraz, Iran. ایران mahmoudvandsh100@yahoo.com سمیه شکری Somayeh Shokri Department of Virology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran. ایران mahmoudvandsh100@yahoo.com کامران زمردیان Kamiar Zomorodian Department of Mycology, Faculty of Medicine, Shiraz University of Medical Sciences Shiraz, Iran. ایران mahmoudvandsh100@yahoo.com Adenovirus Nested-PCR Rhinosinusitis [1]. Meltzer EO, Hamilos DL, Hadley JA, Lanza DC, Marple BF, Nicklas RA, et al. Rhinosinusitis: establishing definitions for clinical research and patient care. J Allergy Clinic Immunol. 2004; 114(6): 155-212.##[2]. Brook I. Microbiology of sinusitis. Proceed Am Thoracic Society. 2011; 8(1): 90-100.##[3]. Pedersen M, Sakakura Y, Winther B, Brofeldt S, Mygind N. Nasal mucociliary transport, number of ciliated cells, and beating pattern in naturally acquired common colds. Euro J Respirat Dis Suppl. 1983; 128(1): 355-65.##[4]. Gwaltney JM, Sydnor A, Sande MA. Etiology and antimicrobial treatment of acute sinusitis. Ann Otol, Rhinol Laryngol Suppl. 1981; 90(3): 68-71.##[5]. Wald ER, Milmoe GJ, Bowen A, Ledesma-Medina J, Salamon N, Bluestone CD. Acute maxillary sinusitis in children. New Eng J Med. 1981; 304(13): 749-54.##[6]. Ghebremedhin B. Human adenovirus: Viral pathogen with increasing importance. Euro J Microbiol Immunol. 2014; 4(1): 26-33.##[7]. Jin Y, Zhang RF, Xie ZP, Yan KL, Gao HC, Song JR, et al. Prevalence of adenovirus in children with acute respiratory tract infection in Lanzhou, China. Virol J. 2013; 10: 271.##[8]. Lynch JP, Kajon AE. Adenovirus: epidemiology, global spread of novel serotypes, and advances in treatment and prevention. Seminars Respirat Critic Care Med. 2016; 37(4): 586-602.##[9]. Hamory BH, Sande MA, Sydnor A, Seale DL, Gwaltney JM. Etiology and antimicrobial therapy of acute maxillary sinusitis. J Infect Dis. 1979; 139(2): 197-202.##[10]. Evans FO, Sydnor JB, Moore WE, Moore GR, Manwaring JL, Brill AH, et al. Sinusitis of the maxillary antrum. New Eng J Med. 1975; 293(15): 735-39.##[11]. Spector SL, English GM, McIntosh K, Farr RS. Adenovirus in the sinuses of an asthmatic patient with apparent selective antibody deficiencies. Am J Med. 1973; 55(2): 227-31.##[12]. Osur SL. Viral respiratory infections in association with asthma and sinusitis: a review. Annals of allergy, asthma & immunology. Official Pub Am College Allergy, Asthma, Immunol. 2002; 89(6): 553-60.##[13]. Cho GS, Moon BJ, Lee BJ, Gong CH, Kim NH, Kim YS, et al. High rates of detection of respiratory viruses in the nasal washes and mucosae of patients with chronic rhinosinusitis. J Clinic Microbiol. 2013; 51(3): 979-84.##[14]. Abshirini H, Makvandi M, Seyyed Ashrafi M, Hamidifard M, Saki N. Prevalence of rhinovirus and respiratory syncytial virus among patients with chronic rhinosinusitis. Jundishapur J microbiol. 2015; 8(3): e20068.##[15]. Allard A, Albinsson B, Wadell G. Rapid typing of human adenoviruses by a general PCR combined with restriction endonuclease analysis. J Clinic Microbiol. 2001; 39(2): 498-505.##[16]. Mahmoudvand S, Safaei A, Erfani N, Sarvari J. Presence of human papillomavirus DNA in colorectal cancer tissues in Shiraz, Southwest Iran. Asian Pac J Cancer Prev. 2015; 16(17): 7883-887.##[17]. Mahmoudvand S, Zamani K, Safaei A, Khashei R, Motamedifar M, Azizi Z, et al. No detection of Streptococcus gallolyticus and Helicobacter pylori in colorectal cancer tissue samples in Shiraz, Iran. Iranian Journal of Cancer Prevention. 2017;10(1): 6337.##[18]. Moattari A, Pirbonyeh N, Yaghoobi R. Detection of Adenovirus infection among children with acute respiratory disease during 2010-2012 in Shiraz, Iran. Archiv Pediatr Infect Dis. 2014; 2(4): 15930.##[19]. Dykewicz MS, Hamilos DL. Rhinitis and sinusitis. J Allergy Clinic Immunol. 2010; 125(S 2): 103-15.##[20]. Wald ER. Epidemiology, pathophysiology and etiology of sinusitis. Pediatric Infect Dis. 1985; 4(S 6): 51-4.##[21]. Aring AM, Chan MM. Acute rhinosinusitis in adults. Am Family Physic. 2011; 83(9): 1057-1063.##[22]. Gwaltney JM, Hendley JO, Phillips CD, Bass CR, Mygind N, Winther B. Nose blowing propels nasal fluid into the paranasal sinuses. Clinical infectious diseases. Official Pub Infect Dis Society Am. 2000; 30(2): 387-91.##[23]. Jin Y, Zhang RF, Xie ZP, Yan KL, Gao HC, Song JR, et al. Prevalence of adenovirus in children with acute respiratory tract infection in Lanzhou, China. Virol J. 2013; 10: 271-77.##[24]. Lim LM, Woo YY, de Bruyne JA, Nathan AM, Kee SY, Chan YF, et al. Epidemiology, clinical presentation and respiratory sequelae of adenovirus pneumonia in children in Kuala Lumpur, Malaysia. PloS one 2018; 13(10): 205795.##[25]. Ramadan HH, Farr RW, Wetmore SJ. Adenovirus and respiratory syncytial virus in chronic sinusitis using polymerase chain reaction. Laryngoscope 1997; 107(7): 923-25.##[26]. Moattari A, Emami A, Pirbonyeh N, Yaghoobi R. Detection of adenovirus infection among children with acute respiratory disease during 2010-2012 in Shiraz, Iran. Archiv Pediat Infect Dis. 2014; 2(4): 1-10.##[27]. Haque E, Banik U, Monowar T, Anthony L, Adhikary AK. Worldwide increased prevalence of human adenovirus type 3 (HAdV-3) respiratory infections is well correlated with heterogeneous hypervariable regions (HVRs) of hexon. PloS one 2018; 13(3): 0194516.## ##

The Effect of Incubation Time on the Activity and Stability of Factor VIII during the Preparation Process 2 2 Background and Aims: Hemophilia is a rare autoimmune disorder caused by autoantibodies directed in the majority of the cases against clotting factor VIII (FVIII). FVIII is extracted from human plasma or engineered from mammalian cell cultures using recombinant DNA technology. In Iran, most of the used FVIII is prepared from human plasma in Iranian Blood Transfusion Organization. It was seen important to estimate its stability and activity from bleeding time until product preparation. Material and Methods: In the analytical study, 60 healthy male donors (20-50 years old), 15 donors from each blood group, were selected after obtaining informed consent. Donors' blood was collected in QUADRI-PACKs and centrifuged after keeping at 24°C for 2 hours. The separated plasma was divided into three groups and incubated in the lab (22°C) for 0, 90, and 180 minutes, respectively. Then, samples were stored at -20°C for one month. Afterward, the plasma was thawed, and FVIII activity was assayed. Results: The activity of FVIII significantly (p<0.05) reduced by delay in freezing; after the time of 0 min: 134.84%±42, after 90 min: 126.88%±38, after 180 min: 120.22%±34. At all incubation times, the highest and the lowest FVIII activity were observed in A and O blood groups, respectively (p<0.05). FVIII activity was increased along with increasing age up to 35-40, but it decreased in subjects of 40-50 years old. These experiments confirmed that the longer the delay in freezing fresh frozen plasma, the greater the decrease in FVIII stability. Conclusions:. According to the results of this study, the best blood donors for FVIII product are those with blood group A in the age range of 40-35 years. 199 206 2019/02/92018/04/162019/03/132019/01/92018/09/252019/05/25 1398/3/4 2019/07/202019/07/152019/07/202019/07/22019/07/22019/07/17 1398/4/26 کوروش کبیر Kourosh Kabir Social Determinants of Health Research Center, Alborz University of Medical Sciences, Karaj, Iran. ایران kabi.korosh@yahoo.com حسن حسینی Hassan Hosseini Blood Transfusion Researcher Center, Institute for Higher Education and Research in Transfusion Medicine, Tehran, Iran. ایران Hosseini.h@yahoo.com مهدی جاهد زرگر Mehdy Jahed Zargar Department of Hematology, Faculty of Paramedical, Alborz University of Medical Sciences, Karaj, Iran. ایران mahdizargar@hotmail.com زینب مانده Zeynab Mandeh Blood Transfusion Researcher Center, Institute for Higher Education and Research in Transfusion Medicine, Tehran, Iran. ایران zeynabmandeh@yahoo.com فاطمه امرالهی Fatemeh Amrollahi Blood Transfusion Researcher Center, Institute for Higher Education and Research in Transfusion Medicine, Tehran, Iran. ایران amrollahi@gmail.com نوید فرهمندیان Navid Farahmandian Department of Biochemistry, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran. ایران farahmandiyan.n@tak.iums.ac.ir الهام بحرینی Elham Bahreini Department of Biochemistry, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran. ایران Bahreini.e@iums.ac.ir Blood Transfusion Factor VIII Hemophilia Incubation time [1]. Wakabayashi H, Freas J, Zhou Q, Fay PJ. Residues 110–126 in the A1 domain of factor VIII contain a Ca2+ binding site required for cofactor activity. J Biol Chem. 2004; 279(13): 12677-2684.##[2]. Fay PJ. Activation of factor VIII and mechanisms of cofactor action. Blood Rev. 2004; 18(1): 1-15.##[3]. Lusher JM, Milestones in hemophilia and concepts in future clinical trial design. Semin Hematol. 2006; 43(1): 83-7.##[4]. Fijnvandraat K, Peters M, Ten Cate JW. Inter‐individual variation in half‐life of infused recombinant factor VIII is related to pre‐infusion von Willebrand factor antigen levels. Br J Haematol. 1995; 91(2): 474-76.##[5]. Franchini M, Mannucci PM. Past, present and future of hemophilia: a narrative review. Orphanet J Rare Dis. 2012; 7(1): 24-29.##[6]. Lynch CM, Israel DI, Kaufman RJ, Miller AD. Sequences in the coding region of clotting factor VIII act as dominant inhibitors of RNA accumulation and protein production. Human Gene Therapy 1993; 4(3): 259-72.##[7]. Lusher JM, Arkin S, Abildgaard CF, Schwartz RS. Recombinant factor VIII for the treatment of previously untreated patients with hemophilia A-safety, efficacy, and development of inhibitors. New Eng J Med. 1993; 328(7): 453-59.##[8]. Dasgupta S, Repessé Y, Bayry J, Navarrete AM, Wootla B, Delignat S, et al. VWF protects FVIII from endocytosis by dendritic cells and subsequent presentation to immune effectors. Blood 2007; 109(2): 610-12.##[9]. Scharrer I, Bray G, Neutzling O. Incidence of inhibitors in haemophilia A patientsa review of recent studies of recombinant and plasma-derived factor VIII concentrates. Haemophilia 1999; 5: 145-54.##[10]. Mannucci PM, Chediak J, Hanna W, Byrnes J, Ledford M, Ewenstein BM, et al. Treatment of von Willebrand disease with a high-purity factor VIII/von Willebrand factor concentrate: a prospective, multicenter study. Blood 2002; 99(2): 450-56.##[11]. Pool JG, Shannon AE. Production of high-potency concentrates of antihemophilic globulin in a closed-bag system: assay in vitro and in vivo. New Eng J Med. 1965; 273(27): 1443-447.##[12]. Rodgers SE, Duncan EM, Barbulescu DM, Quinn DM, Lloyd JV. In vitro kinetics of factor VIII activity in patients with mild haemophilia A and a discrepancy between one‐stage and two‐stage factor VIII assay results. Br J Haematol. 2007; 136(1): 138-45.##[13]. Carlebjörk G, Oswaldsson U, Rosén S. A simple and accurate microplate assay for the determination of factor VIII activity. Thrombos Res. 1987; 47(1): 5-14.##[14]. Smith JF, Ness PM, Moroff G, Luban NL. Retention of coagulation factors in plasma frozen after extended holding at 1-6 C. Vox Sanguinis. 2000; 78(1): 28-30.##[15]. Swärd‐Nilsson AM, Persson PO, Johnson U, Lethagen S. Factors influencing factor VIII activity in frozen plasma. Vox sanguinis 2006; 90(1): 33-9.##[16]. Franchini M, Capra F, Targher G, Montagnana M, Lippi G. Relationship between ABO blood group and von Willebrand factor levels: from biology to clinical implications. Thrombos J. 2007; 5(1): 14.##[17]. Gallinaro L, Cattini MG, Sztukowska M, Padrini R, Sartorello F, Pontara E, et al. A shorter von Willebrand factor survival in O blood group subjects explains how ABO determinants influence plasma von Willebrand factor. Blood 2008; 111(7): 3540-545.##[18]. Gill JC, Endres-Brooks J, Bauer PJ, Marks WJ, Montgomery RR. The effect of ABO blood group on the diagnosis of von Willebrand disease. Blood 1987; 69(6): 1691-695.##[19]. O'donnell J, Laffan M. The relationship between ABO histo‐blood group, factor VIII and von Willebrand factor. Transf Med. 2001; 11(4): 343-51.##[20]. Song J, Chen F, Campos M, Bolgiano D, Houck K, Chambless LE, et al. Quantitative influence of abo blood groups on factor viii and its ratio to von willebrand factor, novel observations from an aric study of 11,673 subjects. PloS one 2015; 10(8): e0132626.##[21]. Smith NL, Chen MH, Dehghan A, Strachan DP, Basu S, Soranzo N, et al. Novel associations of multiple genetic loci with plasma levels of factor VII, factor VIII, and von Willebrand factor. Circulation 2010; 121(12): 1382-392.##[22]. Wang Z, Dou M, Du X, Ma L, Sun P, Cao H, et al. Influences of ABO blood group, age and gender on plasma coagulation factor VIII, fibrinogen, von Willebrand factor and Adamts13 levels in a Chinese population. Peer J. 2017; 5: 3156.##[23]. Cohen W, Castelli C, Alessi MC, Aillaud MF, Bouvet S, Saut N, et al. ABO blood group and von Willebrand factor levels partially explained the incomplete penetrance of congenital thrombophilia. Arterioscleros, Thrombos, Vascul Biol. 2012; 32(8): 2021-2028.##[24]. Gonskikh Y, Polacek N. Alterations of the translation apparatus during aging and stress response. Mech Ageing Dev. 2017; 168(5): 30-6.## ##

Use of Mesenchymal Adult Stem Cell for Cartilage Regeneration by Hydrogel تمایز سلولهای بنیادی بالغ به غضروف با استفاده از داربست زیستی مهندسی بافت غضروف 2 1 زمینه و هدف: غضروف یک بافت بسیار تخصصی است که قابلیت ترمیم خود را در مواقع آسیب ندارد. اگر چه روشهای جراحی با پیوند کندروسیت اتولوگ، روشهایی توسعه یافته برای درمان ضایعات غضروفی هستند، اما نمی توانند در مواجهه با مشکلات ناشی از آسیب ناحیه پیوندی موفقیتهای بزرگی حاصل نمایند. با پیشرفت در زمینه مهندسی بافت و دانش استفاده از سلول های بنیادی مزانشیمی مشتق از چربی و کشت و تمایز آنها در داربست های مناسب به عنوان استراتژی موفق بالینی در نظر گرفته میشوند. مواد و روشها: در این مطالعه پس از آماده سازی دو داربست چسب فیبرینی و آلژینات، سلول های بنیادی مزانشیمی جدا شده از بافت چربی جداگانه بر روی این دو داربست قرار گرفتند و در محیط کندروژنی کشت داده شدند. 1، 7 و 14 روز پس از تمایز سلولی، توانایی زنده ماندن سلول های تمایز یافته توسط MTT کندروژنی مورد تجزیه و تحلیل قرار گرفت. همچنین بیان کلاژن نوع I،نوع II و SOX9 و اگریکان به روش Real time PCr مورد بررسی قرار گرفت. همچنین تشکیل غضروف ها نیز به لحاظ بافت شناسی مورد ارزیابی قرار گرفت. یافته ها: بر اساس نتایج حاصل از این مطالعه، داربست چسب فیبرینی با داربست آلژینات در سلول های تمایزی اختلاف معنی داری داشت. با توجه به تجزیه و تحلیل MTT و علاوه بر این، چسب فیبرینی بالاترین بیان در ژن های کندروژنیک را در میان دیگر داربست ها بود. نتیجه گیری: با توجه به نتایج این مطالعه میتوان چنین عنوان نمود که استفاده از داربست های طبیعی مانند چسب فیبرینی میتواند همانند پروتئین های مزانشیمی سلول های بنیادی به عنوان استراتژی کارآمد و جدید در مهندسی بافت و ترمیم عمل نماید. 2 Background and Aims: Cartilage is a very specific tissue, which does not have the capacity to heal and renew itself. Although the invention of the method of surgery with autologous chondrocyte transplantation, developed tools to treat the cartilage lesions, it couldn’t gain a great success due to problems such as damage to the area of donation. Using the mesenchymal stem cells derived from adipose and culturing and differentiating them on scaffolds was considered appropriate as a successful research and clinical strategy. Materials and Methods: In the present study, the mesenchymal stem cells were separated from adipose tissue and cultured in two scaffolds of fibrin glue and alginate medium. After 1, 7 and 14 days of cell differentiation, the survival ability of the differentiated cells were analyzed by Chondrogenic MTT. Moreover, type I and II collagen, aggrecan and Sox9 expression were measured via real time-polymerase chain reaction. In addition, cartilage reconstruction on scaffolds was shown by a histological investigation. Results: Our results showed that the expression of CD90 and CD105 as mesenchymal markers is at a high level whereas the expression of CD34 and CD45 reaches a low level. The LSD test demonstrated that there was no remarkable difference among the chondrogenic MTT, scaffolds groups and control in 7 and 14 days after cell differentiation (p<0.05), although, fibrin glue had the highest expression in chondrogenic gens. Conclusions: Finding suggests that in order to utilize a new strategy for tissue regeneration utilization of inherent scaffolds such as fibrin glue can act as a protector for mesenchymal stem cells. 207 218 2019/02/92018/04/162019/03/132019/01/92018/09/252019/05/252018/07/13 1397/4/22 2019/07/202019/07/152019/07/202019/07/22019/07/22019/07/172019/07/2 1398/4/11 مهدیه قیاثی Mahdieh Ghiasi Razi Drug Research Center, Iran University of Medical Sciences, Tehran, Iran. ایران mahdieh.ghiasi@yahoo.com محمد مهدی زاده Mohammad Mehdizadeh Dental and Oral Research Center, Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Qom University of Medical Sciences, Qom, Iran ایران علی محمدشریفی Ali Mohammadsharifi Razi Drug Research Center, Iran University of Medical Sciences, Tehran, Iran. ایران sharifal@yahoo.com غلامعباس نصر Gholamabbas Nasr Eurasia University of Science and Research, International Campus, Tehran, Iran ایران Abb13404@gmail.com محمد بیگدلو Mohammad Bigdelo Department of Surgery, Qom University of Medical Sciences, Qom, Iran. ایران Jhandark2010@yahoo.com سحر فرزانه Sahar Farzaneh Department of Biology, Islamic Azad University, Parand Branch, Parand, Iran. ایران Sahar.farzaneh@gmail.com فرزانه تفویضی Farzaneh Tafvizi Department of Biology, Islamic Azad University, Parand Branch, Parand, Iran. ایران صدیقه صفری Sedigheh Safari Department of Biology, Islamic Azad University, Tehran Science and Research Branch, Tehran, Iran . ایران ندا تکیه معروف Neda Tekiyeh Maroof Department of Molecular Biology, Islamic Azad Medical University of Tehran, Tehran, Iran. ایران Adipose tissue Alginates Fibrin tissue adhesive Mesenchymal stem cell سلول های بنیادی مزانشیمی بافت چربی چسب فیبرینی آلژینات [1]. Esteves Vieira Branquinho M, Caseiro AR, Santos Pedrosa S, Damásio Alvites R, Colette Maurício A. Synovia-derived mesenchymal stem cell application in musculoskeletal injuries: a review. Tissue Regenera. 2018; 6: 77.##[2]. Baugé C, Boumédiene K. Use of adult stem cells for cartilage tissue engineering: current status and future developments. Stem Cells Int. 2015; 2015: 438026. ##[3]. Francis SL, Duchi S, Onofrillo C, Di Bella C, Choong PFM. Adipose-derived mesenchymal stem cells in the use of cartilage tissue engineering: the need for a rapid isolation procedure. Stem Cells Int. 2018; 2018: 8947548. ##[4]. Estes BT, Diekman BO, Gimble JM, Guilak F. Isolation of adipose-derived stem cells and their induction to a chondrogenic phenotype. Nat Protoc. 2010; 5: 1294-311.##[5]. Im GI, Shin YW, Lee KB. Do adipose tissue-derived mesenchymal stem cells have the same osteogenic and chondrogenic potential as bone marrow-derived cells? Osteoarthritis Cartilage 2005; 13(10): 845-53.##[6]. Ullah I, Subbarao RB, Rho GJ. Human mesenchymal stem cells-current trends and future prospective. Biosci Rep. 2015; 35(2): 191. ##[7]. Duan, Bin, Hockaday LA, Das S, Xu C, Butcher JT. Comparison of mesenchymal stem cell source differentiation toward human pediatric aortic valve interstitial cells within 3d engineered matrices. Tissue Eng. 2015; 21 (8): 795-807. ##[8]. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2002; 13: 4279-295.##[9]. Vinatier C, Bouffi C, Merceron C, Gordeladze J, Brondello JM, Jorgensen C, et al. Cartilage tissue engineering: towards a biomaterial-assisted mesenchymal stem cell therapy. Curr Stem Cell Res Ther. 2009; 4(4): 318-29.##[10]. Cao N. Fabrication of alginate hydrogel scaffolds and cell viability in calcium-crosslinked alginate hydrogel [Master of Science Thesis]. Biomedical Engineering Department, Saskatchewan University; 2011. ##[11]. Awad HA, Wickham MQ, Leddy HA, Gimble JM, Guilak F. Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds. Biomaterials 2004; 25(16): 3211-222.##[12]. Ahmed TA, Dare EV, Hincke M .Fibrin: a versatile scaffold for tissue engineering applications. Tissue Eng Part B Rev 2008; 14(2): 199-215.##[13]. Ahmed TA, Griffith M, Hincke M. Characterization and inhibition of fibrin hydrogel-degrading enzymes during development of tissue engineering scaffolds. Tissue Eng. 2007; 13(7): 1469-477.##[14]. Ahmed TA, Halpenny M, Atkins H, Giulivi A, Dervin G, Griffith M, et al. Stabilization of fibrinmesenchymal stem cells (MSCs) constructs under hypoxic conditions during tissue engineering of articular cartilage. J Bone Joint Surg Br. 2010; 92(2): 2-3.##[15]. Park JS, Shim MS, Shim SH, Yang HN, Jeon SY, Woo DG, et al. Chondrogenic potential of stem cells derived from amniotic fluid, adipose tissue, or bone marrow encapsulated in fibrin gels containing TGF-beta3. Biomaterials 2011; 32(32): 8139-849.##[16]. Diederichs S, Baral K, Tanner M, Richter W. Interplay between local versus soluble TGF-beta and fibrin scaffolds: role of cells and impact on human mesenchymal stem cell chondrogenesis. Tissue Eng. 2012; 18(2): 1140-150.##[17]. Ahmed TA, Giulivi A, Griffith M, Hincke M. Fibrin glues in combination with mesenchymal stem cells to develop a tissue-engineered cartilage substitute. Tissue Eng. 2011; 17(3-4): 323-35.##[18]. Jung SN, Rhie JW, Kwon H, Jun YJ, Seo JW, Yoo G, et al. In vivo cartilage formation using chondrogenic-differentiated human adipose-derived mesenchymal stem cells mixed with fibrin glue. J Craniofac Surg. 2010; 21(2):468-72.##[19]. Yang HN, Park JS, Woo DG, Jeon SY, Do HJ, Lim HY, et al. Chondrogenesis of mesen-chymal stem cells and dedifferentiated chondro-cytes by transfection with SOX Trio genes. Biomaterials 2011; 32(30):7695-704.##[20]. Li Z, Kupcsik L, Yao SJ, Alini M, Stoddart MJ. Chondrogenesis of human bone marrow mesenchymal stem cells in fibrin-polyurethane composites. Tissue Eng. 2009; 15(2):1729-737.##[21]. Li Z, Yao SJ, Alini M, Stoddart MJ. Chond-rogenesis of human bone marrow mesenchymal stem cells in fibrin-polyurethane composites is modulated by frequency and amplitude of dynamic compression and shear stress. Tissue Eng. 2010; 16(3): 575-84.##[22]. Barry F, Boynton RE, Liu B, Murphy JM. Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. Exp Cell Res. 2001; 268(2): 189-200.##[23]. Ho ST, Cool SM, Hui JH, Hutmacher DW. The influence of fibrin based hydrogels on the chondrogenic differentiation of human bone marrow stromal cells. Biomaterials 2010; 31(1): 38-47.##[24]. Zanetti AS, Sabliov C, Gimble JM, Hayes DJ. Human adipose-derived stem cells and three-dimensional scaffold constructs: A review of the biomaterials and models currently used for bone regeneration. Journal of Biomedical Materials Research. Applied Biomaterials 2013; 101(1): 187-99.##[25]. Sheykhhasan M, Qomi RT, Kalhor N, Mehdizadeh M, Ghiasi M. Evaluation of the ability of natural and synthetic scaffolds in providing an appropriate environment for growth and chondrogenic differentiation of adipose-derived mesenchymal stem cells. Indian J Orthop. 2015; 49(5): 561-68.##[26]. Sheykhhasan M, Qomi RT, Ghiasi M. Fibrin scaffolds designing in order to human adipose-derived mesenchymal stem cells differentiation to chondrocytes in the presence of TGF-β3. Int J Stem Cell 2015; 8(2): 219-27.##[27]. Ghiasi M, Kalhor N, Tabatabaei Qomi R, Sheykhhasan M. The effects of synthetic and natural scaffolds on viability and proliferation of adipose-derived stem cells. Frontiers Life Sci. 2016; 9(1): 32-43.##[28]. Tigli RS, Cannizaro C, Gumusderelioglu M, Kaplan DL. Chondrogenesis in perfusion bioreactors using porous silk scaffolds and hESC-derived MSCs. J Biomed Mater Res. 2011; 96(1): 21-8.##[29]. Proulx MK, Carey SP, DiTroia LM, Jones C, Fakharzadeh M, Guyette JP, et al. Fibrin microthreads support mesenchymal stem cell growth while maintaining differentiation potential. J Biomed Mater Res. 2011; 96(5): 301-12.##[30]. Chaudhari AA, Vig K, Baganizi D, Sahu R, Dixit S, Dennis V, et al. Future prospects for scaffolding methods and biomaterials in skin tissue engineering: a review. Int J Mol Sci. 2016; 17(12): 1974.##[31]. Focaroli S, Teti G, Salvatore V, Orienti I, Falconi M. Calcium/cobalt alginate beads as functional scaffolds for cartilage tissue engineering. Stem Cells Int. 2017; 2(5): 2374-381.##[32]. Zhang L, Hu J, Athanasiou KA. The role of tissue engineering in articular cartilage repair and regeneration. Crit Rev Biomed Eng. 2009; 37(1-2): 1-57.##[33]. Ahmed TA, Hincke MT. Strategies for articular cartilage lesion repair and functional restoration. Tissue Eng Rev. 2010; 16(3): 305-29.## ##

Effects of Aqueous and Ethanolic Extracts of Myrtus Communis Leaves on Trophozoites and Cysts of Acanthamoeba: An In Vitro Study 2 2 Background and Aims: Acanthamoeba is a ubiquitous amphizoic organism which can cause lethal diseases such as granulomatous amoebic encephalitis and unfortunately, the infection has now increased in the world. The aim here was to evaluate in vitro anti-Acanthamoeba properties of crude aqueous and ethanolic extracts of Myrtus communis. Materials and Methods: In this experimental research, a clinical isolate of Acanthamoeba was cultured and genotyped. The aqueous and ethanolic extracts of Myrtus communis were prepared. Then, various concentrations of Myrtus communis extracts (1.25, 2.5, 5, and 10 mg/ml) were tested at three different times (24, 48 and 72 hr) on trophozoites and cysts of Acanthamoeba in vitro. The viability of trophozoites or cysts was tested by trypan blue method. Unstained (viable) and stained (nonviable) parasites were evaluated by counting with a neobar lam. Results: The percentage of viablity of trophozoites and cysts after adding ethanolic extract of Myrtus communis was 0% and 8.62%, respectively. Moreover, at 10 mg/ml concentration of aqueous extract of Myrtus communis, 0% trophozoites and 31.10% cysts lived after 72 h. Conclusions: This extract can be used as a safe anti-Acanthamoeba agent against trophozoites and cysts of Acanthamoeba and further investigations are recommended to show the effects of this plant as an antiparasitic drug in animal models and volunteer infected people. 219 225 2019/02/92018/04/162019/03/132019/01/92018/09/252019/05/252018/07/132018/12/13 1397/9/22 2019/07/202019/07/152019/07/202019/07/22019/07/22019/07/172019/07/22019/07/2 1398/4/11 توران نیری چگینی Tooran Nayeri Chegeni Department of Parasitology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran. ایران tooran.nayeri@ modares.ac.ir فاطمه غفاریفر Fatemeh Ghaffarifar Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران ghafarif@modares.ac.ir فریبا خوشزبان Fariba Khoshzaban Department of Parasitology, Shahed University of Medical Sciences, Tehran, Iran. ایران fkhosh_99@yahoo.com عبدالحسین دلیمی اصل Abdolhosein Dalimi Asl Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران dalimi_a@modares.ac.ir هدی میرزائیان Hoda Mirzaian Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران hoda.mirzaian@yahoo.com فرنوش جامعی Farnoosh Jameie Department of Parasitology, Razi Vaccine and Serum Reaserch Institute, Agricultural Reaserch, Edjucation and Extension Organization (AREEO), Karaj, Alborz, Iran. ایران f.jameie@ modares.ac.ir Acanthamoeba Keratitis In vitro Myrtus communis [1]. 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