The Requisiteness for not Sacrificing Medical Biotechnology in the Coronavirus Era 2 2 The coronavirus storm, first reported more than a year ago, has overshadowed all societies' parts and become a challenge to all of the world's health systems 1. Extremely high contagiousness, significant mortality rates, and the lack of a definitive cure have prioritized overcoming this outbreak. In this regard, studies related to coronavirus and especially its clinical studies, became a priority for researchers and decision-makers at the request of governments and the people, as well as by the logic. The superiority of an emergency is not a wrong decision. Still, the problem arose when other research areas were neglected, and their budgets were reduced by decision-makers, resulting in damage to the research and researchers in other fields 2,3. The clinical studies currently being conducted on coronavirus disease 2019 (COVID-19) are due to previous studies in the basic sciences that have provided the background. Obviously, without scientists' tasks over the years and the allocation of research funds to the fields of genetics, biochemistry, immunology, etc., studies and advances would not be possible today 4,5. Medical biotechnology is a type of applied science that produces or creates products that improve human health, mainly through genetic engineering and tissue culture, using biological systems or living organisms 5,6. However, this area of science can also be harmful through studies' unintended consequences, the production of products without genetic diversity, and deliberate biological manipulation. In medical biotechnology, using basic sciences such as biochemistry, biology, and genetics, and by modifying cells or cell subsets, the prevention of diseases-including the production of vaccines-and the treatment of diseases, especially by creating novel agents, are studied 6-10. It should be noted that the background of these lofty goals has been years of basic science studies, many of which have not reached a positive conclusion or have been rejected by more recent research. Even they more often lead to more questions instead of answers 4. In other words, the passing of many years and spending on scientific and research projects has enabled human beings today to create advanced products for fighting pathogens and improving society's health. The development of insulin, the production of advanced monoclonal antibodies, and vaccines' production against an RNA virus are some of the notable novel products 6,7. Since the beginning of the coronavirus outbreak, many ways have been suggested to prevent and treat the disease. Today, after more than a year and the reported deaths of nearly two million people from COVID-19, the highest hopes for overcoming the disorder are with the proposed vaccines 1,11. Science achieves patients' treatment through basic studies 4, so the right decision is a decision that, while meeting the need, does not disrupt the long-established science and research system. We can see that the time and money spent in the past is helping all sections of society today with the production of the coronavirus vaccine, and if those studies had not been done in the past for whatever reason, today we were a few steps behind. Likewise, suppose today, for any reason, even the allocation of all time and budget to the emergency situation, the pace of progress in this area slows down. 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Using CRISPR/Cas9 System to Knock out Exon 48 in DMD Gene 2 2 Background: Out of frame mutations in DMD gene cause Duchenne Muscular Dystrophy (DMD) which is a neuromuscular progressive genetic disorder. In DMD patients, lack of dystrophin causes progressive muscle degeneration, which results in heart and respiratory failure leading to premature death. At present, there is no certain treatment for DMD. DMD gene is the largest gene in human genome by 2.2 mega base pairs and contains 79 exons. In the past few years, gene therapy has been considered a promising DMD treatment, and among various gene-editing technologies, CRISPR/Cas9 system is shown to be more precise and reliable. The aim of this study was to assess the possibility of knocking out exon 48 by using a pair of sgRNAs. Methods: A pair of guide RNAs (gRNAs) was designed to cleave DMD gene and induce deletion of exon 48. gRNAs were transfected to the HEK-293 cell line and then the deletion in genomic DNA was analyzed by PCR and subsequent Sanger sequencing. Results: Exon 48 was successfully deleted and therefore exon 47 was joined to exon 49. Conclusion: This result indicated that CRISPR/Cas9 system could be used to edit DMD gene precisely. 54 57 Mahintaj Dara Department of Molecular Medicine, Shiraz University of Medical Sciences Department of Molecular Medicine, Shiraz University of Medical Sciences Iran Vahid Razban Department of Molecular Medicine, Shiraz University of Medical Sciences Department of Molecular Medicine, Shiraz University of Medical Sciences Iran Mahdieh Talebzadeh Department of Molecular Medicine, Shiraz University of Medical Sciences Department of Molecular Medicine, Shiraz University of Medical Sciences Iran Sepideh Moradi Department of Molecular Medicine, Shiraz University of Medical Sciences Department of Molecular Medicine, Shiraz University of Medical Sciences Iran Mehdi Dianatpour Department of Medical Genetics, Shiraz University of Medical SciencesStem Cell Technology Research Center, Shiraz University of Medical Sciences Department of Medical Genetics, Shiraz University of Medical SciencesStem Cell Technology Research Center, Shiraz University of Medical Sciences IranIran CRISPR/Cas9DystrophinGene editingMuscular dystrophies 40450.pdf Nelson CE, Gersbach CA. Genome editing for Duchenne muscular dystrophy. Mus Gen Thera 2019:383-403.##Falzarano MS, Scotton C, Passarelli C, Ferlini A. Duchenne muscular dystrophy: from diagnosis to therapy. Molecules 2015;20(10):18168-84.##Mias-Lucquin D, Morais RDS, Chéron A, Lagarrigue M, Winder SJ, Chenuel T, et al. How the central domain of dystrophin acts to bridge F-actin to sarcolemmal lipids. J Struc Biol 2020;209(1):107411.##Wang L, Xu M, Li H, Zhu Y, He R, Lin J, et al. Genotypes and phenotypes of DMD small mutations in chinese patients with dystrophinopathies. Front Genet 2019;10:114.##Schreiber A, Brochard S, Rippert P, Fontaine‐Carbonnel S, Payan C, Poirot I, et al. Corticosteroids in Duchenne muscular dystrophy: impact on the motor function measure sensitivity to change and implications for clinical trials. Dev Med Child Neurol 2018;60(2):185-91.##Savić N, Schwank G. Advances in therapeutic CRISPR/Cas9 genome editing. Trans Res 2016;168:15-21.##O’Brien A, Cohn RD. Genome editing for muscle gene therapy. Mus Gen Thera 2019;275-87.##Moretti A, Fonteyne L, Giesert F, Hoppmann P, Meier A, Bozoglu T, et al. Somatic gene editing ameliorates skeletal and cardiac muscle failure in pig and human models of Duchenne muscular dystrophy. Nat Med 2020;26(2):207-14.##López-Hernández LB, Gómez-Díaz B, Luna-Angulo AB, Anaya-Segura M, Bunyan DJ, Zúñiga-Guzman C, et al. Comparison of mutation profiles in the duchenne muscular dystrophy gene among populations: Implications for potential molecular therapies. Int J Mol Sci 2015;16(3):5334-46.##Iskandar K, Dwianingsih EK, Pratiwi L, Kalim AS, Mardhiah H, Putranti AH, et al. The analysis of DMD gene deletions by multiplex PCR in Indonesian DMD/BMD patients: the era of personalized medicine. BMC Res Notes 2019;12(1):704. ##Vieitez I, Gallano P, González-Quereda L, Borrego S, Marcos I, Millán J, et al. Mutational spectrum of Duchenne muscular dystrophy in Spain: Study of 284 cases. Neurología 2017;32(6):377-85.##Min YL, Li H, Rodriguez-Caycedo C, Mireault AA, Huang J, Shelton JM, et al. CRISPR-Cas9 corrects Duchenne muscular dystrophy exon 44 deletion mutations in mice and human cells. Sci Adv 2019;5(3):eaav4324.##Tabebordbar M, Zhu K, Cheng JKW, Chew WL, Widrick JJ, Yan WX, et al. In vivo gene editing in dystrophic mouse muscle and muscle stem cells. Science 2016;351(6271):407-11. ##Barzegar M, Habibi P, Bonyady M, Topchizadeh V, Shiva S. Exon deletion pattern in duchene muscular dystrophy in north west of Iran. Iran J Child Neurol 2015;9(1):42-8. ##Shariati Gh, Shakerian S, Anaie MM, Abdorasouli N, Nanvazadeh F, Sedaghat A, et al. Deletion and duplication mutations spectrum in Duchenne muscular dystrophy in the southwest of Iran. Meta Gene 2020;23:100641.##

Designing and Development of a Tandem Bivalent Nanobody against VEGF165 2 2 Background: Inhibition of angiogenesis using monoclonal antibodies is an effective strategy in cancer therapy. However, they could not penetrate sufficiently into solid tumors. Antibody fragments have solved this issue. However, they suffer from short in vivo half-life. In the current study, a tandem bivalent strategy was used to enhance the pharmacokinetic parameters of an anti-VEGF165 nanobody. Methods: Homology modeling and MD simulation were used to check the stability of protein. The cDNA was cloned into pHEN6C vector and the expression was investigated in WK6 Escherichia coli (E. coli) cells by SDS-PAGE and western blot. After purification, the size distribution of tandem bivalent nanobody was investigated by dynamic light scattering. Moreover, in vitro antiproliferative activity and pharmacokinetic study were studied in HUVECs and Balb/c mice, respectively. Results: RMSD analysis revealed the tandem bivalent nanobody had good structural stability after 50 ns of simulation. A hinge region of llama IgG2 was used to fuse the domains. The expression was induced by 1 mM IPTG at 25°C for overnight. A 30 kDa band in 12% polyacrylamide gel and nitrocellulose paper has confirmed the expression. The protein was successfully purified using metal affinity chromatography. MTT assay revealed there is no significant difference between the antiproliferative activity of tandem bivalent nanobody and the native protein. The hydrodynamic radius and terminal half-life of tandem bivalent nanobody increased approximately 2-fold by multivalency compared to the native protein. Conclusion: Our data revealed that the physicochemical as well as in vivo pharmacokinetic parameters of tandem bivalent nanobody was significantly improved. 58 64 Farnaz Khodabakhsh Department of Genetics and Advanced Medical Technology, Medical Biotechnology Research Center, Faculty of Medicine, AJA University of Medical Sciences Department of Genetics and Advanced Medical Technology, Medical Biotechnology Research Center, Faculty of Medicine, AJA University of Medical Sciences Iran Morteza Salimian Department of Medical Laboratory, Kashan University of Medical Sciences Department of Medical Laboratory, Kashan University of Medical Sciences Pardis Ziaee Department of Biology, Central Tehran Branch, Islamic Azad University Department of Biology, Central Tehran Branch, Islamic Azad University Iran Fatemeh Kazemi-Lomedasht Venom and Biotherapeutics Molecules Laboratory, Department of Biotechnology, Biotechnology Research Center, Pasteur Institute of Iran Iran Mahdi Behdani Reza Ahangari Cohan CancerPharmacokineticsSingle domain antibodyVascular endothelial growth factor 40451.pdf Kijanka M, Dorresteijn B, Oliveira S, van Bergen en Henegouwen PM. 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Methylation of TGM-3 Promoter and Its Association with Oral Squamous Cell Carcinoma (OSCC) 2 2 Background: Oral Squamous Cell Carcinoma (OSCC) is among the ten most common cancers worldwide. Hypermethylation of CpG sites in the promoter region and subsequent down-regulation of a tumor suppressor gene, TGM-3 has been proposed to be linked to different types of human cancers including OSCC. In this study, methylation status of CpG sites in the promoter region of TGM-3 has been evaluated in a cohort of patients with OSCC compared to normal controls. Methods: Forty fresh tissue samples were obtained from newly diagnosed OSCC patients and normal individuals referred to dentistry clinic for tooth extraction. DNA was extracted, bisulfite conversion was performed and it was subjected to PCR using bisulfite-sequencing PCR (BSP) primers. Prepared samples were sequenced on a DNA analyzer with both forward and reverse primers of the region of interest. The peak height values of cytosine and thymine were calculated and methylation levels for each CpG site within the DNA sequence was quantified. Results: Quantitative DNA methylation analyses in CpG islands revealed that it was significantly higher in OSCC patients compared to controls. DNA methylation at CpG1/CpG3/CpG5 (p=0.004-0.01) and CpG1/CpG3 (p=0.001-0.019) sites was associated with tumor stage and grade, respectively. Male OSCC patients had higher methylation rate at CpG3 (p=0.032), while smoker patients showed higher methylation rate at CpG6 (p=0.045). Conclusion: These results manifested the contribution of DNA methylation of TGM-3 in OSCC and its potential association with clinico-pathologic parameters in OSCC. 65 73 Sorour Shojaeian Department of Biochemistry, Alborz University of Medical Sciences Department of Biochemistry, Alborz University of Medical Sciences Iran Abdolkarim Moazeni-Roodi Department of Clinical Biochemistry, Iranshahr University of Medical Sciences Department of Clinical Biochemistry, Iranshahr University of Medical Sciences Iran Abdolamir Allameh epartment of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University epartment of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University Iran Ata Garajei Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Tehran University of Medical SciencesDepartment of Head and Neck Surgical Oncology and Reconstructive Surgery, The Cancer Institute, Faculty of Medicine, Tehran University of Medical Sciences Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Tehran University of Medical SciencesDepartment of Head and Neck Surgical Oncology and Reconstructive Surgery, The Cancer Institute, Faculty of Medicine, Tehran University of Medical Sciences IranIran Anoshirvan Kazemnejad Department of Bio-statistics, Faculty of Medical Sciences, Tarbiat Modares University Department of Bio-statistics, Faculty of Medical Sciences, Tarbiat Modares University Iran Kourosh Kabir Department of Community Medicine, Alborz University of Medical Sciences Department of Community Medicine, Alborz University of Medical Sciences Iran Amir-Hassan Zarnani Department of Immunology, Faculty of Public Health, Tehran University of Medical SciencesReproductive Immunology Research Center, Avicenna Research Institute, ACECR Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences IranIran DNA methylationGeneticOral squamous cell carcinomaPromoter regions 40452.pdf Scully C. 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Developmental Toxicity of the Neural Tube Induced by Titanium Dioxide Nanoparticles in Mouse Embryos 2 2 Background: This study investigated the potential effects of Titanium dioxide nanoparticles (Tio2NPs) followed by maternal gavage on fetal development and neural tube formation during pregnancy in mice. Methods: Thirty pregnant mice were randomly divided into five main study groups including the untreated control and 4 experimental groups (n=6 per group). The control group was treated with normal saline and the experimental groups were orally treated with doses of 30, 150, 300, and 500 mg/kg Body Weight (BW) of Tio2NPs during pregnancy. On gestational day 16 and 19 (n=3 per group), pregnant mice were euthanized and then examined for neural tube defects and compared with control. Serial transverse sections were prepared in both cranial region and in lumbar region of spinal cord. Results: Treatment with Tio2NPs resulted in low fetal weight and short length, dilation of lateral ventricle, thinning of cerebral cortex and spinal cord, spina bifida occulta and an increase in the number of apoptotic neurons in exposed embryos at doses of 300 and 500 mg/kg (p<0.05). Conclusion: It seems that exposure to nanoparticles of Tio2 during pregnancy induces growth retardation and for the first time, teratogenicity of this nanomaterial in neural tube development and induction of defects such as spinal bifida, reduction in cortical thickness and  dilatation of lateral ventricles were verified which can be related to incidence of apoptosis in central nervous system. 74 80 Nahid Mohamadzadeh Department of Anatomical Sciences, Faculty of Medicine, Urmia University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Urmia University of Medical Sciences Iran Masoumeh Zirak Javanmard Mojtaba Karimipour Department of Anatomical Sciences, Faculty of Medicine, Urmia University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Urmia University of Medical Sciences Iran Gholamhosain Farjah Department of Anatomical Sciences, Faculty of Medicine, Urmia University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Urmia University of Medical Sciences Iran Fetal developmentMiceNeural tube defectsTitanium dioxide 40453.pdf Ybot-Gonzalez P, Copp AJ. 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Reproductive toxicity provoked by titanium dioxide nanoparticles and the ameliorative role of Tiron in adult male rats. Biochem Biophys Res Commun 2017;486(2):595-600. ##Yamashita K, Yoshioka Y, Higashisaka K, Mimura K, Morishita Y, Nozaki M. Silica and titanium dioxide nanoparticles cause pregnancy complications in mice. Nat Nanotechnol 2011;6(5):321-8. ##Chen IC, Hsiao IL, Lin HC, Wu CH, Chuang CY, Huang YJ. Influence of silver and titanium dioxide nanoparticles on in vitro blood-brain barrier permeability. Environ Toxicol Pharmacol 2016;47:108-18. ##Di Bona KR, Xu Y, Ramirez PA, DeLaine J, Parker C, Bao Y, et al. Surface charge and dosage dependent potential developmental toxicity and biodistribution of iron oxide nanoparticles in pregnant CD-1 mice. Reprod Toxicol 2014;50:36-42. ##Philbrook NA, Winn LM, Afrooz AN, Saleh NB, Walker VK. The effect of TiO2 and Ag nanoparticles on reproduction and development of Drosophila melanogaster and CD-1 mice. 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The Effect of Sodium Selenite on Expression of Mitochondrial Transcription Factor A during In Vitro Maturation of Mouse Oocyte 2 2 Background: The aim of the present study was to investigate the effect of Sodium Selenite (SS) supplemented media on oocyte maturation, expression of mitochondrial transcription factor A (TFAM) and embryo quality. Methods: Mouse Germinal Vesicle (GV) oocytes were collected after administration of Pregnant Mare Serum Gonadotropin (PMSG); in experimental group 1, oocytes were cultured and then subjected for in vitro maturation in the absence of SS, and in experimental group 2, they were matured in vitro in the presence of 10 ng/ml of SS up to 16 hr. The control group included MII oocytes obtained from the fallopian tubes after ovarian stimulation with PMSG, followed by human chorionic gonadotropin. Then, the expression of TFAM in MII oocytes in all three groups was investigated using real-time RT-PCR. The fertilization and embryo developmental rates were assessed, and finally the quality of the blastocysts was evaluated using propidium iodide staining. Results: The oocyte maturation rate to MII stage in SS treated group was significantly higher than non-treated oocytes (75.65 vs. 68.17%, p<0.05). Also, the rates of fertilization, embryo development to blastocyst stage as well as the cell number of blastocyst in SS supplemented group were higher than other experimental group (p<0.05). There was a significant decrease in TFAM gene expression in both in vitro groups compared to the group with in vivo obtained oocytes (p<0.05). Moreover, there was a significant increase in TFAM gene expression in oocytes that matured in the presence of SS compared to that of the group without SS (p<0.05). Conclusion: Supplementation of oocyte maturation culture media with SS improved the development rate of oocytes and embryo and also enhanced TFAM expression in MII oocytes which can affect the mitochondrial biogenesis of oocytes. 81 86 Tina Moshaashaee Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Iran Saeed Zavareh Faculty of Biology, Damghan University Faculty of Biology, Damghan University Iran Shahram Pourbeiranvand Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University Iran Mojdeh Salehnia Gene expressionIn vitro oocyte maturationMiceMitochondrial transcription factor AOocytesSodium selenite 40454.pdf Hatirnaz S, Ata B, Hatirnaz ES, Dahan MH, Tannus S, Tan J, et al. Oocyte in vitro maturation: A sytematic review. J Turkish Soc Obstet Gynecol 2018;15:112-25. ##Costello MF, Garad RM, Hart R, Homer H, Johnson L, Jordan C, et al. A review of second- and third-line infertility treatments and supporting evidence in women with polycystic ovary syndrome. Med Sci (Basel) 2019;7(7):75. ##Abbara A, Clarke SA, Dhillo WS. Novel concepts for inducing final oocyte maturation in in vitro fertilization treatment. Endocr Rev 2018;39(5):593-628. ##Creux H, Monnier P, Son WY, Buckett W. Thirteen years' experience in fertility preservation for cancer patients after in vitro fertilization and in vitro maturation treatments. J Assist Reprod Genet 2018;35(4):583-92. ##Son WY, Henderson S, Cohen Y, Dahan M, Buckett W. Immature oocyte for fertility preservation. Front Endocrinol 2019;10:464. ##Sato A, Sarentonglaga B, Ogata K, Yamaguchi M, Hara A, Atchalalt K, et al. Effects of insulin-like growth factor-1 on the in vitro maturation of canine oocytes. J Reprod Devel 2018;64(1):83-8. ##Alvarez GM, Barrios Exposito MJ, Elia E, Paz D, Morado S, Cetica PD. Effects of gonadotrophins and insulin on glucose uptake in the porcine cumulus-oocyte complex during IVM. Reprod Fertil Dev 2019. ##Asaadi A, Kafi M, Atashi H, Azari M, Hostens M. Frozen-thawed ampullary cell monolayer improves bovine embryo in vitro development and quality. Zygote 2019;27(5):337-46. ##Heydarnejad A, Ostadhosseini S, Varnosfaderani SR, Jafarpour F, Moghimi A, Nasr-Esfahani MH. Supplementation of maturation medium with CoQ10 enhances developmental competence of ovine oocytes through improvement of mitochondrial function. Mol Reprod Devel 2019;86(7):812-24. ##John JC. Mitochondria and female germline stem cells, a mitochondrial DNA perspective. Cells 2019;8(8):852. ##Schatten H, Sun QY, Prather R. The impact of mitochondrial function/dysfunction on IVF and new treatment possibilities for infertility. Reprod Biol Endocrinol 2014;12:111. ##Kasapoglu I, Seli E. Mitochondrial dysfunction and ovarian aging. Endocrinology 2020;161(2):bqaa001. ##Ghorbanmehr N, Salehnia M, Amooshahi M. The effects of sodium selenite on mitochondrial DNA copy number and reactive oxygen species levels of in vitro matured mouse oocytes. Cell J 2018;20(3):396-402. ##Sharma N, Pasala MS, Prakash A. Mitochondrial DNA: epigenetics and environment. Environ Mol Mutagen 2019;60(8):668-2. ##Farge G, Falkenberg M. Organization of DNA in mammalian mitochondria. Int J Mol Sci 2019;20(11):2770. ##Ngo HB, Lovely GA, Phillips R, Chan DC. Distinct structural features of TFAM drive mitochondrial DNA packaging versus transcriptional activation. Nat Commun 2014;5:3077. ##Ghaffari Novin M, Noruzinia M, Allahveisi A, Saremi A, Fadaei Fathabadi F, Mastery Farahani R, et al. Comparison of mitochondrial-related transcriptional levels of TFAM, NRF1 and MT-CO1 genes in single human oocytes at various stages of the oocyte maturation. Iran Biomed J 2015;19(1):23-8. ##Nie J, Yan K, Sui L, Zhang H, Zhang H, Yang X, et al. Mogroside V improves porcine oocyte in vitro maturation and subsequent embryonic development. Theriogenology 2020;141:35-40. ##Dai X, Qiu L, Zhao B, Gao Y, Mu Y, Chu Z, et al. Melatonin ameliorates the fertilization capacity of oocytes exposed to 17alpha-ethynylestradiol. Reprod Toxicol 2020;93:61-7. ##Venditti P, Meo SD. The role of reactive oxygen species in the life cycle of the mitochondrion. Int J Mol Sci 2020;21(6):2173. ##Sena LA, Chandel NS. Physiological roles of mitochondrial reactive oxygen species. Mol Cell 2012;48(2):158-67. ##Yang S, Lian G. ROS and diseases: role in metabolism and energy supply. Mol Cell Biochem 2020;467(1-2):1-12. ##Xiong X, Lan D, Li J, Lin Y, Li M. Selenium supplementation during in vitro maturation enhances meiosis and developmental capacity of yak oocytes. Anim Sci J 2018;89(2):298-306. ##Lizarraga RM, Anchordoquy JM, Galarza EM, Farnetano NA, Carranza-Martin A, Furnus CC, et al. Sodium selenite improves in vitro maturation of bos primigenius taurus oocytes. Biol Trace Elem Res 2020;197(1):149-58. ##Malekzadeh M, Salehnia M, Zavareh S. Effect of sodium selenite on the apoptotic genes expression in in vitro matured mouse metaphase II oocyte. Veter Res Forum 2020:(In press).##Amoushahi M, Salehnia M, Ghorbanmehr N. The mitochondrial DNA copy number, cytochrome c oxidase activity and reactive oxygen species level in metaphase II oocytes obtained from in vitro culture of cryopreserved ovarian tissue in comparison with in vivo-obtained oocyte. J Obstet Gynaecol Res 2018;44(10):1937-46. ##Ghaemi SR, Salehnia M, Valojerdi MR. The effect of progesterone and exogenous gonadotropin on preimplantation mouse embryo development and implantation. Exp Anim 2008;57(1):27-34. ##Abedelahi A, Salehnia M, Allameh AA, Davoodi D. 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Sodium selenite inhibits deoxynivalenol-induced injury in GPX1-knockdown porcine splenic lymphocytes in culture. Sci Rep 2018;8(1):17676. ##Colle D, Santos DB, de Souza V, Lopes MW, Leal RB, de Souza Brocardo P, et al. Sodium selenite protects from 3-nitropropionic acid-induced oxidative stress in cultured primary cortical neurons. Mol Biol Rep 2019;46(1):751-62. ##Yang L, Cai YS, Xu K, Zhu JL, Li YB, Wu XQ, et al. Sodium selenite induces apoptosis and inhibits autophagy in human synovial sarcoma cell line SW982 in vitro. Mol Med Rep 2018;17(5):6560-8. ##Piantadosi CA, Suliman HB. Mitochondrial transcription factor A induction by redox activation of nuclear respiratory factor 1. J Biol Chem 2006;281(1):324-33. ##Battaglia R, Vento ME, Ragusa M, Barbagallo D, La Ferlita A, Di Emidio G, et al. MicroRNAs are stored in human MII oocyte and their expression profile changes in reproductive aging. Biol Reprod 2016;95(6):131. ##

Benchmarking Datasets from Malaria Cytotoxic T-cell Epitopes Using Machine Learning Approach 2 2 Background: Epitope prediction remains a major challenge in malaria due to the unique parasite biology, in addition to rapidly evolving parasite sequence variation in Plasmodium species. Although several models for epitope prediction exist, they are not useful in Plasmodium specific epitope development. Hence, it was proposed to use machine learning based methods to develop a peptide sequence based epitope predictor specific for malaria. Methods: Model datasets were developed and performance was tested using various machine learning algorithms. Machine learning classifiers were trained on epitope data using sequence features and comparison of amino acid physicochemical properties was done to yield a valid prediction model. Results: The findings from the analysis reveal that the model developed using selected classifiers after preprocessing by Waikato Environment for Knowledge Analysis (WEKA) performed better than other methods. The datasets for benchmarks of performance are deposited in the repository https://github.com/githubramaadiga/epito-pe_dataset. Conclusion: The study is the first in-silico study on benchmarking Plasmodium cytotoxic T cell epitope datasets using machine learning approach. The peptide based predictors have been used for the first time to classify cytotoxic T cell epitopes in malaria. Algorithms has been evaluated using real datasets from malaria to obtain the model. 87 91 Rama Adiga BenchmarkingEpitopesMachine learningMalariaPlasmodium 40455.pdf Angermueller C, Pärnamaa T, Parts L, Stegle O. Deep learning for computational biology. Mol Syst Biol 2016;12(7):878. ##Riihimäki H, Chachólski W, Theorell J, Hillert J, Ramanujam R. 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Designing Two Synthetic Constructs for Real Time PCR Detection of Francisella tularensis and Ebola Virus 2 2 Background: Generally, timely diagnosis of micro-organisms is very important to prevent many diseases. Many methods can detect micro-organisms like culture-based methods and molecular methods. The molecular methods are usually preferred because they provide fast and reliable results. In some cases, microbial strains are not accessible, and there is no safety to work with them; therefore, synthetic constructs which are designed according to the available sequences in databases can be used as a positive control for detection of them. Methods: In this study, a synthetic construct was designed for molecular detection of Francisella tularensis (F. tularensis) and the Ebola virus by multiplex real-time PCR reaction. For this, sequences were taken from databases and then multiple alignments were done by software. Also, conventional PCR and two models of real-time PCR (SYBR green and TaqMan) were applied. Finally, multiplex real-time PCR was performed. Results: The synthetic construct was designed and used for conventional PCR and multiplex PCR. The results of common PCR showed a single band at 148 bp and 167 bp in 1.5% agarose gel stained by ethidium bromide for F. tularensis and Ebola virus, respectively. Also, a dual-band at 148 and 167 bp was observed in multiplex PCR. Results of real-time PCR showed a limit of detection about 0.1 pg of plasmid/µl. Conclusion: In conclusion, the designed construct can be used as a positive control for an accurate diagnosis of these micro-organisms without any biological danger for laboratory staff. So, this method is useful for diagnosis of these agents in food, water, and blood samples. 92 97 Mohammadjavad Dehghan Esmat Abadi Hesam Motalebzadeh Department of Medical Biotechnology, Faculty of Allied Medical Sciences, Iran University of Medical Sciences Department of Medical Biotechnology, Faculty of Allied Medical Sciences, Iran University of Medical Sciences Iran Mahmoud Barati Department of Genetics, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University Department of Genetics, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University Iran Mohammadali Yaghobi Faculty of Chemistry and Chemical Engineering, Malek Ashtar University of Technology Faculty of Chemistry and Chemical Engineering, Malek Ashtar University of Technology Iran Ebola virusHumansMultiplex polymerase chain reactionReal time polymerase chain reaction 40456.pdf Baden LR, Kanapathipillai R, Campion EW, Morrissey S, Rubin EJ, Drazen JM. Ebola--an ongoing crisis. 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The Effect of Silver Nanoparticles on Pyocyanin Production of Pseudomonas aeruginosa Isolated From Clinical Specimens 2 2 Background: Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen causing a wide range of human infections. The organism is resistant to a wide range of antibiotics. The purpose of this study was to investigate the effect of AgNPs on pyocyanin pigment production of P. aeruginosa bacteria isolated from clinical specimens. Methods: In this study, 15 clinical isolates of P. aeruginosa were collected from different specimens of hospitalized patients. P. aeruginosa was detected by biochemical and molecular (detection of pbo1 gene by colony PCR method) methods and the MIC and MBC of AgNPs were determined by agar dilution method. Inhibition of P. aeruginosa pyocyanin production at AgNPs concentrations of 0, 0.3, 0.5, 1 and 1.5 mg/ml of was studied with OD of 520 nm. Results: The mean MIC and MBC of AgNPs were 1.229 and 1.687 mg/ml, respectively. Pyocyanin production was investigated for all isolates at different concentrations of nanoparticles, and their comparison showed that with increasing nanoparticle concentration, pyocyanin production significantly decreased (p<0.05). Conclusion: According to the results of this study, AgNPs had an inhibitory effect on P. aeruginosa and its pigment production and with increasing nanoparticles concentration, pigment production decreased; therefore, it seems that the nanoparticles can be used to treat and prevent diseases caused by P. aeruginosa. 98 103 Mahboobeh Najafi Department of Biology, Faculty of Science, Damghan Branch, Islamic Azad University Department of Biology, Faculty of Science, Damghan Branch, Islamic Azad University Iran Mahboobeh Nakhaei Moghaddam Department of Biology, Faculty of Science, Mashhad Branch, Islamic Azad University Department of Biology, Faculty of Science, Mashhad Branch, Islamic Azad University Iran Ehsan Yousefi NanoparticlesPolymerase chain reaction Pseudomonas aeruginosaPyocyanin 40457.pdf Hoseinzadeh E, Samarghandi MR, Alikhani MY, Asgari G, Roshanaei GHA. Effect of zinc-oxide nanoparticles on death kinetic of gram-negative and positive bacterium. 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