Coronavirus Vaccination and Mortality in the Omicron Outbreak in Iran: Mortality Reduction due to Attenuated Pathogenicity and Booster Vaccine Doses 2 2 More than two years after the first report of the rapidly spreading coronavirus disease 2019 (COVID-19), the coronavirus is still unpredictable by emerging highly divergent variants 1. Through the highly contagious viral sickness the virus causes, it imposes significant morbidity and mortality on global populations, becoming the second cause of death in people aged 25 to 44 years in January 2021 in the United States. Although mortality rates in disease peaks were significantly reduced as vaccination rates increased, mortality increased again with the outbreak of a variant of concern called Omicron so that the virus became the second leading cause of death in January 2022 in the United States 1,2. The novel variant, first identified in Botswana and named Omicron (B.1.1.529) by World Health Organization on November 26, 2021, exhibited 36 mutations in the spike protein, the target of antibodies, and a total of 59 mutations throughout its genome. More importantly, 15 mutations occurred in the Receptor-Binding Domain (RBD), which can increase infectivity and mediate virus escape from vaccine-induced antibodies 3. In other words, the highly mutated omicron variant can evade immunity in vaccinated individuals and is associated with vaccine breakthroughs 1,3. The Omicron has a 13-fold increase in viral infectivity, and compared to the Delta variant, it is 2.8 times more contagious and its R0, the indicator of contagiousness and transmissibility, can be as high as 10, while for the Delta, it did not reach 7 4. Fortunately, the mortality of this variant is lower than the wild type and Alpha, Beta and Delta variants 5. About two years after the first official report of the SARS-CoV-2 outbreak in Iran, like in other countries, the virus is still raging. As in other countries, the peak of the Omicron variant in Iran increased infection, hospitalization, and mortality once again. During the Omicron outbreak in Iran, the maximum number of newly identified cases in one day was more than 39,000, so that the maximum number of active cases in one day reached about 375,000 by mid-February 2022. Although in the previous peak, the maximum number of new cases detected in a day and the maximum number of deaths per day were about 50,000 and more than 700, respectively, the number of deaths per day did not reach 250 in this peak 6. This difference may be due to the lower mortality5 and progress of the vaccination process, although vaccine efficacy is negatively affected by new variants 3. Although the percentages of people who have received at least one dose of a vaccine, have been fully vaccinated, and given the booster dose are now about 75%, 65%, and 25%, respectively, these percentages on August 24, 2021, when the highest number of deaths due to COVID-19 were reported in Iran, were about 20%, 7%, and 0, respectively 7. The fastest and most accessible ways to protect against the Omicron variant seem to be wearing medical-grade masks and receiving the third dose of vaccine 8. The protection provided by two shots of vaccines is reduced to less than 40% a few months after the second shot, but the third shot seems to cause about 60 to 70% protection in the two weeks after the injection and protect against severe COVID-19 9. It has been shown that vaccine efficacy decreases with increasing time after vaccination 3. There is not much difference in the treatment of the disease caused by this variant, but of the monoclonal antibodies, only sotrovimab appears to be effective and is authorized 4. Eventually, updating the current vaccines and pursuing a universal vaccine should be of concern to those in charge 8,10. 102 103 Ahmad Shamabadi School of Medicine, Tehran University of Medical SciencesPsychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences School of Medicine, Tehran University of Medical SciencesPsychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences IranIran Shahin Akhondzadeh Psychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences Psychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences Iran Coronavirus spike proteinCOVID-19 PandemicMass active immunizationSARS-CoV-2 60492.pdf Kannan S, Shaik Syed Ali P, Sheeza A. 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Cellular Therapy: The Hope for Covid-19 2 2 Coronaviruses (CoVs) are a group of very diverse viruses that cause a broad spectrum of diseases from mild to severe enteric, respiratory, systemic diseases, and common cold or pneumonia among humans and animals. This virus is associated with Middle East Respiratory Syndrome (MERS), Severe Acute Respiratory Syndrome (SARS), and lung disease that lead to Acute Respiratory Distress Syndrome (ARDS). In December 2019, researchers identified a novel coronavirus type, called Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2), which was associated with symptoms of high fever, dry cough, headache, diarrhea, and reduction of White Blood Cells (WBC). Coronavirus-associated acute respiratory disease was named Coronavirus Disease 19 (COVID-19). No proven treatment has been discovered for COVID-19 so far, but researchers are trying to find the best effective way to treat this disease. Therefore, therapeutic strategies that facilitate the recovery of COVID-19 patients and reduce life-threatening complications are urgently needed now. Today, Mesenchymal Stem Cells (MSCs) and their secretion are utilized as one of the most applied tools to treat various diseases such as inflammation and cancer. MSC-derived vesicles are rich in various growth factors, cytokines, and interleukins that are produced and secreted under different physiological or pathological conditions. These vesicles were considered a suitable and effective tool in regeneration medicine because of their high power in repairing damaged tissues and modulating immune responses. Recently, evidence has shown MSC-derived vesicles through reduced expression of pro-inflammatory cytokines could improve damaged tissues in COVID-19 patients. In addition to MSCs and MSC-derived exosomes, Natural Killer (NK) cells, T cells, and platelet lysates were used against viral infection. In this review, we tried to provide an overview of MSC secretion and immune cells for COVID-19 therapy. 104 113 Sima Nobari Research Center for Molecular Medicine, School of Medicine, Hamadan University of Medical Sciences Research Center for Molecular Medicine, School of Medicine, Hamadan University of Medical Sciences Iran Motahareh Rezvan Department of Photo Healing and Regeneration, Medical Laser Research Center, Yara Institute, ACECR Iran Fariba Dashtestani Institute of Biochemistry and Biophysics, University of Tehran Institute of Biochemistry and Biophysics, University of Tehran Iran Mahdieh Gangi Department of Biology, Payame Noor University Department of Biology, Payame Noor University Iran Hoda Keshmiri Neghab Department of Photo Healing and Regeneration, Medical Laser Research Center, Yara Institute, ACECR Iran COVID-19Mesenchymal stem cell Natural killer cellsPlatelet lysesSARS-CoV-2 60493.pdf Mastri M, Lin H, Lee T. Enhancing the efficacy of mesenchymal stem cell therapy. 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Designing a Multi-Epitope Antigen for Serodiagnosis of Strongyloides stercoralis Based on L3Nie.01 and IgG Immunoreactive Epitopes 2 2 Background: Serological diagnosis of Strongyloides stercoralis  (S. stercoralis) is fre-quently challenging because of cross-reactivity with other parasitic nematodes. There-fore, it is necessary to introduce novel serological tests with high performance to properly diagnose this neglected parasitic infection. The purpose of the current study was to design a multi-epitope construct for the diagnosis of S. stercoralis. Methods: For the purpose of this study, first, highly antigenic segments and potential immunodominant epitopes of S. stercoralis were identified from two antigenic proteins, and then all of the selected parts were linked by an appropriate linker. Next, the physico-chemical features of the designed construct were analyzed. Then, tertiary structures of the construct were built and evaluated to find out the best one. Lastly, the amino acid sequence was reverse-translated and optimized for over-expression in Escherchia coli (E. coli). Results: The bioinformatic evaluation indicated that the designed protein construct could be hydrophilic, thermostable, and acidic and the estimated half-life was more than 10 hr in E. coli. Conclusion: According to the results of the study, the designed construct could be used as an efficient antigen in the ELISA system for serological diagnosis of human strong-yloidiasis. 114 124 Ahmad Movahedpour Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical SciencesStudent Research Committee, Shiraz University of Medical Sciences Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical SciencesStudent Research Committee, Shiraz University of Medical Sciences IranIran Zohreh Mostafavi-Pour Recombinant Protein Laboratory, Department of Biochemistry, School of Medicine, Shiraz University of Medical SciencesAutophagy Research Center, Shiraz University of Medical Sciences Recombinant Protein Laboratory, Department of Biochemistry, School of Medicine, Shiraz University of Medical SciencesAutophagy Research Center, Shiraz University of Medical Sciences IranIran Bahador Sarkari Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical SciencesBasic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical SciencesBasic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences IranIran Mortaza Taheri-Anganeh Cellular and Molecular Research Center, Cellular and Molecular Medicine Institute, Urmia University of Medical Sciences Cellular and Molecular Research Center, Cellular and Molecular Medicine Institute, Urmia University of Medical Sciences Iran Navid Nezafat Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences Iran Amir Savardashtaki Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences Infertility Research Center, Shiraz University of Medical Sciences Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences IranIran Younes Ghasemi Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences Iran AntigensMulti-epitopesSerological diagnosisStrongyloides stercoralis 60494.pdf Casavechia MTG, Lonardoni MVC, Venazzi EAS, Cam-panerut-Sá PAZ, da Costa Benalia HR, Mattiello MF, et al. 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CpG-Containing Oligodeoxynucleotides and Freund Adjuvant in Combination with Alum Augment the Production of Monoclonal Antibodies Against Recombinant HBsAg 2 2 Background: Adjuvants are essential to potentiate the immune response to inoculated antigens and play a central role in vaccine development. Alum is generally used as a classic adjuvant, although it does not stimulate proper immunity, and some of the immunized subjects have low or no antibody response. Efforts have been continued to find more efficient adjuvants for better antibody responses. In the present study, the efficacy of three formulations of adjuvants, i.e. Cysteine p Guanine Oligodeoxynucleotide (CpG ODN), alum, and Freund, in the production of monoclonal anti Hepatitis B Surface Antigen (HBsAg) antibodies was investigated. Methods: To immunize mice, regular hepatitis B vaccine containing recombinant HBsAg and alum was used with CpG ODN or Freund adjuvants, and splenocytes of hyperimmunized mice were fused with murine myeloma Sp2/0 cells. Positive hybridomas were selected by Enzyme-Linked Immunosorbent Assay (ELISA) using HBsAg as coating antigen followed by a limited dilution process. Results: The results showed that by using all three formulations of adjuvants, monoclonal antibody (mAb) specific to HBsAg was successfully generated. It was also found that the mice immunized with (HBsAg + Alum) + CpG had the highest concentration of antibody production in serum and hybridoma supernatants as well as positive clones. Based on these findings, the addition of CpG ODN also induced a higher antibody response compared with Complete Freund’s Adjuvant (CFA). 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The Inhibitory Effect of Thioridazine on adeB Efflux Pump Gene Expression in Multidrug-Resistant Acinetobacter baumannii Isolates Using Real Time PCR 2 2 Background: The purpose of the present study was to investigate the antimicrobial effects of berberine and thioridazine, as well as their effect on the gene expression of the AdeABC efflux pump system in Multidrug-Resistant (MDR) Acinetobacter baumannii (A. baumannii) isolates. Methods: This study was carried out in five MDR clinical isolates of A. baumannii and a sample of standard strain (A. baumannii PTCC1797). The effect of Minimum Inhibitory Concentration (MIC) and the Minimum Bactericidal Concentration (MBC) of berberine, thioridazine, and ciprofloxacin alone and their combination on A. baumannii was evaluated by broth microdilution method. Also, their effect on the expression of adeB efflux pump gene was evaluated using real time PCR method.        Results: The MIC of thioridazine, berberine, ciprofloxacin+thioridazine, ciprofloxacin+ berberine, thioridazine+berberine, and ciprofloxacin+thioridazine+berberine on MDR A. baumannii isolates was 64, 256, 128, 256, 128, and 128 μg/ml, respectively. The results showed that treatment of strains with thioridazine alone and in combination with berberine and ciprofloxacin significantly (p<0.05) decreased the expression of adeB efflux pump gene. Conclusion: Due to the inhibitory effects of thioridazine on bacterial isolates and adeB efflux pump gene, this compound can be used as a potential antimicrobial agent against MDR A. baumannii. 132 136 Fereshteh Ahmadi Student Research Committee, Yasuj University of Medical Sciences Student Research Committee, Yasuj University of Medical Sciences Iran Bahman Khalvati Medicinal Plants Research Center, Yasuj University of Medical Sciences Medicinal Plants Research Center, Yasuj University of Medical Sciences Iran Saba Eslami Shiraz University of Medical Sciences Shiraz University of Medical Sciences Iran Mehdi Mirzaii Faculty of Medicine, Shahroud University of Medical Sciences Faculty of Medicine, Shahroud University of Medical Sciences Iran Narges Roustaei Department of Epidemiology and Biostatistics, School of Health and Nutrition Sciences, Social Determinants of Health Research Center, Yasuj University of Medical Sciences Department of Epidemiology and Biostatistics, School of Health and Nutrition Sciences, Social Determinants of Health Research Center, Yasuj University of Medical Sciences Iran Farzad Mazloomirad Student Research Committee, Yasuj University of Medical Sciences Student Research Committee, Yasuj University of Medical Sciences Iran Seyed Sajjad Khoramrooz Cellular and Molecular Research Center, Yasuj University of Medical SciencesDepartment of Microbiology, School of Medicine, Yasuj University of Medical Sciences Cellular and Molecular Research Center, Yasuj University of Medical SciencesDepartment of Microbiology, School of Medicine, Yasuj University of Medical Sciences IranIran Acinetobacter baumanniiCiprofloxacinGene expressionReal time polymerase chain reactionThioridazine 60496.pdf Galac MR, Snesrud E, Lebreton F, Stam J, Julius M, Ong AC, et al. A diverse panel of clinical Acinetobacter bau-mannii for research and development. Antimicrob Agents Chemother 2020;64(10):e00840-20. ##Behdad R, Pargol M, Mirzaie A, Karizi SZ, Noor-bazargan H, Akbarzadeh I. Efflux pump inhibitory activity of biologically synthesized silver nanoparticles against multidrug‐resistant Acinetobacter baumannii cli-nical isolates. J Basic Microbiol 2020;60(6):494-507. ##Nowak J, Schneiders T, Seifert H, Higgins PG. The Asp20-to-Asn substitution in the response regulator AdeR leads to enhanced efflux activity of AdeB in Acinetobacter baumannii. Antimicrob Agents Chemother 2016;60(2):1085-90. ##Xu C, Bilya S, Xu W. adeABC efflux gene in Aci-netobacter baumannii. New Microbes New Infect 2019; 30:100549. ##Jin J, Hua G, Meng Z, Gao P. Antibacterial mechanisms of berberine and reasons for little resistance of bacteria. Chin Herb Med 2010;3:27-35.##Mahmoudi H, Zare Fahim N, Alikhani MY, Shokoohi-zadeh L. Investigation of antimicrobial effect of ber-berine on ciprofloxacin and imipenem resistance Acine-tobacter baumannii isolated from Hamadan hospitals. Iranian J Med Microbiol 2020;14(1):44-54.##Luan W, Liu X, Wang X, An Y, Wang Y, Wang C, et al. Inhibition of drug resistance of Staphylococcus aureus by efflux pump inhibitor and autolysis inducer to strengthen the antibacterial activity of β-lactam drugs. Polish J Microbiol 2019;68(4):477-91. ##Patel JB. Performance standards for antimicrobial sus-ceptibility testing: Clinical and Laboratory Standards Institute; 2017.##Schwalbe R, Steele-Moore L, Goodwin AC. Antimicro-bial susceptibility testing protocols. CRC Press; 2007. 432 p.##Moghadasi M, Kalantar-Neyestanaki D, Rahdar H, Jasemi S, Karami-Zarandi M, Feizabadi MM. Investi-gation of antimicrobial susceptibility patterns and fre-quency of bla OXA genes in carbapenem resistant Acinetobacter baumannii strains. Scientific Journal of Kurdistan University of Medical Sciences 2018;23(5).##Hornsey M, Ellington MJ, Doumith M, Thomas CP, Gordon NC, Wareham DW, et al. AdeABC-mediated efflux and tigecycline MICs for epidemic clones of Acinetobacter baumannii. J Antimicrob Chemother 2010; 65(8):1589-93##Abdi SN, Ghotaslou R, Asgharzadeh M, Mehramouz B, Hasani A, Baghi HB, et al. AdeB efflux pump gene knockdown by mRNA mediated peptide nucleic acid in multidrug resistance Acinetobacter baumannii. Microb Pathog 2020;139:103825. ##Ruzin A, Immermann FW, Bradford PA. RT-PCR and statistical analyses of adeABC expression in clinical iso-lates of Acinetobacter calcoaceticus–Acinetobacter bau-mannii complex. Microb Drug Resist 2010;16(2):87-9. ##Jørgensen NS, Saaby L, Andersson AM, Kromann S, Sheikhsamani E, Permin A, et al. A novel derivative of thioridazine shows low toxicity and efficient activity against Gram-positive pathogens. Antibiotics (Basel) 2020;9(6):327. ##Mahmoudi H, Shokoohizadeh L, Fahim NZ, Bardebari AM, Moradkhani S, Alikhani MY. Detection of adeABC efllux pump encoding genes and antimicrobial effect of Mentha longifolia and Menthol on MICs of imipenem and ciprofloxacin in clinical isolates of Acinetobacter baumannii. BMC Complement Med Ther 2020;20(1):1-7. ##Aghayan SS, Mogadam HK, Fazli M, Darban-Sarokhalil D, Khoramrooz SS, Jabalameli F, et al. The effects of ber-berine and palmatine on efflux pumps inhibition with dif-ferent gene patterns in Pseudomonas aeruginosa iso- lated from burn infections. Avicenna J Med Biotechnol 2017;9(1):2-7.##Gao WW, Gopala L, Bheemanaboina RRY, Zhang GB, Li S, Zhou CH. Discovery of 2-aminothiazolyl berberine derivatives as effectively antibacterial agents toward cli-nically drug-resistant Gram-negative Acinetobacter bau-manii. Eur J Med Chem 2018;146:15-37. ##

A Motif-Based Network Analysis of Regulatory Patterns in Doxorubicin Effects on Treating Breast Cancer, a Systems Biology Study 2 2 Background: Breast cancer is the most common malignancy worldwide. Doxorubicin is an anthracycline used to treat breast cancer as the first treatment choice. Never-theless, the molecular mechanisms underlying the response to Doxorubicin and its side effects are not comprehensively understood so far. We used systems biology and bio-informatics methods to identify essential genes and molecular mechanisms behind the body response to Doxorubicin and its side effects in breast cancer patients. Methods: Omics data were extracted and analyzed to construct the protein-protein interaction and gene regulatory networks. Network analysis was performed to iden-tify hubs, bottlenecks, clusters, and regulatory motifs to evaluate crucial genes and molecular mechanisms behind the body response to Doxorubicin and its side effects. Results: Analyzing the constructed PPI and gene-TF-miRNA regulatory network showed that MCM3, MCM10, and TP53 are key hub-bottlenecks and seed proteins. Enrichment analysis also revealed cell cycle, TP53 signaling, Forkhead box O (FoxO) signaling, and viral carcinogenesis as essential pathways in response to this drug. Besides, SNARE interactions in vesicular transport and neurotrophin signaling were identified as pathways related to the side effects of Doxorubicin. The apoptosis in-duction, DNA repair, invasion inhibition, metastasis, and DNA replication are sug-gested as critical molecular mechanisms underlying Doxorubicin anti-cancer effect. SNARE interactions in vesicular transport and neurotrophin signaling and FoxO signaling pathways in glucose metabolism are probably the mechanisms responsible for side effects of Doxorubicin. Conclusion: Following our model validation using the existing experimental data, we recommend our other newly predicted biomarkers and pathways as possible mole-cular mechanisms and side effects underlying the response to Doxorubicin in breast cancer requiring further investigations. 137 153 Zeinab Dehghan Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences IranIran Seyed Amir Mirmotalebisohi Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences IranIran Marzieh Sameni Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences IranIran Maryam Bazgiri Department of Animal Science, Agriculture and Natural Resources University of Khuzestan Department of Animal Science, Agriculture and Natural Resources University of Khuzestan Iran Hakimeh Zali Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences Iran Breast cancerDoxorubicinProtein-protein interaction networkRegulatory motifSystems biology 60497.pdf Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. 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Bioinformatic Investigation of Micro RNA-802 Target Genes, Protein Networks, and Its Potential Prognostic Value in Breast Cancer 2 2 Background: An increasing number of studies have suggested that unveiling the molecular network of miRNAs may provide novel therapeutic targets or biomarkers. In this study, we investigated the probable molecular functions that are related to microRNA-802 (miR-802) and evaluated its prognostic value in breast cancer utilizing bioinformatics tools. Methods: PPI network, pathway enrichment and transcription factor analysis were applied to obtain hub genes among overlapping genes of four miRNA target prediction databases. Prognosis value assessments and expression analysis of hub genes using bioinformatics tools, as well as their literature validation were performed.  Results: Our results showed a significant correlation of the miR-802 overexpression with poor patient survival rate (BC, p=2.7e-5). We determined 247 target genes significant for GO and KEGG terms. Analysis of TFs by TRUST showed that RUNX3, FOXO3, and E2F1 are possible TFs that regulate the miR-802 expression and target genes network. According to our analysis; 21 genes might have an important function in miR-802 molecular processes and regulatory networks. The result shows that among these 21 genes, 8 genes (CASC3, ITGA4, AGO3, TARDBP, MED13L, SF1, SNRPE and CRNKL1) are positively correlated with patient survival. Therefore these genes could be considered and experimentally evaluated as a prognostic biomarker for breast cancer. Conclusion: The comprehensive bioinformatics study on miR-802 target genes provided insight into miR-802 mediated pathways and processes. Furthermore, representing candidate target genes by prognostic values indicates the potential clinical application of miR-802 in breast cancer. 154 164 Maryam Eini Department of Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences Department of Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences Iran Sepideh Parsi Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School WorcesterDepartment of Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School WorcesterDepartment of Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences USAIran Mahmoud Barati Pasteur Institute of Iran Iran Golnaz Bahramali Pasteur Institute of Iran Iran Marziyeh Alizadeh Zarei Gametogenesis Research Center, Kashan University of Medical Sciences Gametogenesis Research Center, Kashan University of Medical Sciences Iran Jafar Kiani Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences Iran Asaad Azarnezhad Liver and Digestive Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences Liver and Digestive Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences Iran Arshad Hosseini Department of Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences Department of Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences Iran BioinformaticsBreast cancerCell cyclemiR-802PrognosisWnt signaling 60498.pdf Ruhl J, Callaghan C, Hurlbut A. 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Association of Aberrant Promoter Methylation Changes in the Suppressor of Cytokine Signaling 3 (SOCS3) Gene with Susceptibility to Crohn's Disease 2 2 Background: Growing evidence supports that changes in the methylation state of Inflammatory Bowel Disease (IBD)-associated genes could significantly alter levels of gene expression, potentially contributing to disease onset and progression. We supposed that alterations in DNA methylation status at promoter region within the suppressor of cytokine signaling 3 (SOCS3) gene in intestinal tissues may be involved in the susceptibility to Crohn's Disease (CD). Methods: DNA methylation status in the promoter region of the human SOCS3 gene of intestinal tissues from 15 patients with CD and 15 age- and sex-matched healthy controls were profiled using the real-time Quantitative Multiplex Methylation Specific PCR (QM-MSP) assay. Results: Based on methylation assay data profiling, we found that patients with CD showed a higher degree of methylation of the SOCS3 gene promoter region than did the healthy controls (unmethylated DNA in CD vs. healthy controls; 0.00048±0.0011 vs. 0.07±0.142, p<0.000). Conclusion: The data presented here demonstrate that aberrant methylation of the CpG islands within promoter regions of SOCS3 gene in colonic mucosa of CD was associated with mucosal inflammatory status, providing insights into the involvement of methylation could contribute to the initiation of the inflammatory process and development of CD. 165 169 Golshid Sanati Department of Immunology, School of Medicine, Tehran University of Medical Sciences Department of Immunology, School of Medicine, Tehran University of Medical Sciences Iran Davood Jafari Department of Immunology, School of Medicine, Zanjan University of Medical SciencesImmunogenetics Research Network (IgReN), Universal Scientific Education and Research Network (USERN) Department of Immunology, School of Medicine, Zanjan University of Medical Sciences IranIran Mehrdad Noruzinia Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Iran Naser Ebrahimi-Daryani Department of Internal Medicine, Division of Gastroenterology, Imam Khomeini Hospital, Tehran University of Medical Sciences Iran Mohammad Ahmadvand Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Iran Shahram Teimourian Department of Genetics, School of Medicine, Iran University of Medical Sciences Department of Genetics, School of Medicine, Iran University of Medical Sciences Iran Nima Rezaei Department of Immunology, School of Medicine, Tehran University of Medical SciencesResearch Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical SciencesNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN) Department of Immunology, School of Medicine, Tehran University of Medical Sciences IranIranUSA Crohn's diseaseDNA methylationEpigeneticImmune regulationSOCS3 60499.pdf Bojic D, Markovic S. 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Strong Association of Polymorphism in SPRED2 Gene with Disease Susceptibility and Clinical Characteristics of Rheumatoid Arthritis in the Iranian Population 2 2 Background: The high heritability of Rheumatoid Arthritis (RA) has been estimated from different studies. Recently, Genome-Wide Association Studies (GWAS) show a large number of Single Nucleotide Polymorphisms (SNPs) loci affecting susceptibility to RA. The rs934734 polymorphism in the SPRED2 gene is one of these loci. Studies have shown that the SPRED2 gene is involved in the regulation of inflammatory response, leukocyte infiltration, and local chemokine production. In the current study, the possible association between SNP rs934734 (intronic variant) in the SPRED2 gene with RA risk in the Iranian population was evaluated. Methods: One hundred fourteen RA patients and 120 healthy counterparts were recruited in this case-control study to evaluate rs934734 genotypes using the real-time PCR High Resolution Melting method (HRM). Results: Logistic regression analysis demonstrated that GG and AG genotypes compared with AA genotype increase the risk of RA (GG vs. AA; OR=4.61; 95%CI [2.21-9.35]; p<0.001 and AG vs. AA; OR=2.54; 95%CI [1.36-4.76]; p=0.004). Furthermore, subjects with allele G were more frequently affected with RA than subjects with A allele (OR=2.33; 95%CI [1.61-3.38]; p<0.001). Besides, in the patient group, there was a significant correlation between Erythrocyte Sedimentation Rate (ESR) and C-reactive protein (CRP) concentration with rs934734 polymorphism (p<0.05). Conclusion: Our findings suggest that rs934734 in SPRED2 strongly underlies RA development and is associated with clinicopathological characteristics of this disease. 170 174 Bahram Pakzad Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Iran Hamed Moghadammanesh Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Iran Mansour Salesi Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Iran Rasoul Salehi Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences Iran Autoimmune diseaseGenotypeIranRheumatoid arthritisSingle nucleotide polymorphisms 60500.pdf Silman AJ, Pearson JE. 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Association of MTHFR, BMP4, TGFA and IRF6 Polymorphisms with Non-Syndromic Cleft lip and Palate in North Indian Patients 2 2 Background: Non-Syndromic Cleft Lip and Palate (NSCL/P) is a multifactorial birth defect. The world-wide prevalence of NSCL/P is 1 in 1000 live births; it differs with race, ethnicity and gender. The aim of the present study was to find out the status of candidate gene polymorphisms in NSCL/P cases and its association in phenotype of the patients. Methods: We have screened five polymorphisms in four candidate genes MTHFR (rs1801133, rs1801131) BMP4 (rs17563), TGFA (rs1146297) and IRF6 (rs2235371) by restriction fragment length polymorphism and results were validated by Sanger sequencing. Our dataset consists of 200 NSCL/P cases and 200 healthy controls from the Indian population. Statistical data analysis was performed by SPSS software. Results: MTHFR (rs1801133), BMP4 (rs175563) and TGFA (rs11466297) gene polymorphisms showed significant association with NSCL/P and act as a risk factor in the Indian population (p=<0.05). However, MTHFR (rs1801131), and IRF6 (rs2235371) gene polymorphisms did not show significant association with NSCL/P in the Indian population. Conclusion: The result of the study suggests an association between MTHFR (rs1801133), BMP4 (rs175563) and TGFA (rs11466297) polymorphisms with NSCL/P in Indian population. 175 180 Kapil Avasthi Department of Medical Genetics, Sanjay Gandhi Postgraduate Institute of Medical Sciences (SGPGIMS) Department of Medical Genetics, Sanjay Gandhi Postgraduate Institute of Medical Sciences (SGPGIMS) India Amit Agarwal Department of Burn and Plastic Surgery, Vivekananda Polyclinic and Institute of Medical Sciences (VPIMS) Department of Burn and Plastic Surgery, Vivekananda Polyclinic and Institute of Medical Sciences (VPIMS) India Sarita Agarwal Department of Medical Genetics, Sanjay Gandhi Postgraduate Institute of Medical Sciences (SGPGIMS) Department of Medical Genetics, Sanjay Gandhi Postgraduate Institute of Medical Sciences (SGPGIMS) India BMP4Indian populationIRF6MTHFRNSCL/PTGFA 60501.pdf Balaji SM. Burden of orofacial clefting in India, 2016: A global burden of disease approach. 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Plast Surg Int 2012;2012:247104. ##Saket M, Saliminejad K, Kamali K, Moghadam FA, Anvar NE, Khorram Khorshid H. R. 2016. BMP2 and BMP4 variations and risk of non-syndromic cleft lip and palate. Arch Oral Biol 2016;72:134-7. ##Vieira AR. Association between the transforming growth factor alpha gene and nonsyndromic oral clefts: a HuGE review. Am J Epidemiol 2006;163(9):790-810. ##Lu XC, Yu W, Tao Y, Zhao PL, Li K, Tang LJ, et al. Contribution of transforming growth factor α polymor-phisms to nonsyndromic orofacial clefts: a HuGE review and meta-analysis. Am J Epidemiol 2014;179(3):267-81.##Ardinger HH, Buetow KH, Bell GI, Bardach J, VanDe-mark DR, Murray JC. Association of genetic variation of the transforming growth factor-alpha gene with cleft lip and palate. Am J Hum Genet 1989;45(3):348-53.##Ebadifar A, Khorram Khorshid HR, Saliminejad K, Kama-li K, Aghakhani Moghadam F, Esmaeili Anvar N, et al. Association of transforming growth factor alpha polymor-phisms with nonsyndromic cleft lip and palate in Iranian population. Avicenna J Med Biotechnol 2015;7 (4):168-72. ##Zucchero TM, Cooper ME, Maher BS, Daack-Hirsch S, Nepomuceno B, Ribeiro L, et al. Interferon regulatory fac-tor 6 (IRF6) gene variants and the risk of isolated cleft lip or palate. N Engl J Med 2004;351(8):769-80. ##Kondo S, Schutte BC, Richardson RJ, Bjork BC, Knight AS, Watanabe Y, et al. Mutations in IRF6 cause Van der Woude and popliteal pterygium syndromes. Nat Genet 2002;32(2):285-9. ##Rahimov F, Marazita ML, Visel A, Cooper ME, Hitchler MJ, Rubini M, et al. Disruption of an AP-2alpha binding site in an IRF6 enhancer is associated with cleft lip. Nat Genet 2008;40(11):1341–7.##Nan X, Liu M, Yuan G. [Zhonghua zheng xing wai ke za zhi Zhonghua zhengxing waike zazhi]. Zhonghua Zheng Xing Wai Ke Za Zhi 2014;30(4):265-9. Chinese. ##Zhao M, Ren Y, Shen L, Zhang Y, Zhou B. Association between MTHFR C677T and A1298C polymorphisms and NSCL/P risk in Asians: a meta-analysis. PloS One 2014;9(3):e88242. ##Mills JL, Kirke PN, Molloy AM, Burke H, Conley MR, Lee YJ, et al. Methylenetetrahydrofolate reductase thermo-labile variant and oral clefts. A m J Med Genet 1999;86(1):71-4.##Murthy J, Gurramkonda VB, Karthik N, Lakkakula BV. MTHFR C677T and A1298C polymorphisms and risk of nonsyndromic orofacial clefts in a south Indian popula-tion. Int J Pediatr Otorhinolaryngol 2014;78(2):339-42. ##Ali A, Singh KS, Raman R. MTHFR 677TT alone and IRF6 820GG together with MTHFR 677CT, but not MTHFR A1298C, are risks for nonsyndromic cleft lip with or without cleft palate in an Indian population. Genet Test Mol Biomarkers 2009;13(3):355-60. ##Wang W, Jiao XH, Wang XP, Sun XY, Dong C. MTR, MTRR, and MTHFR gene polymorphisms and suscepti-bility to nonsyndromic cleft lip with or without cleft palate. Genet Test Mol Biomarkers 2016;20(6):297-303. ##Sözen MA, Tolarova MM, Spritz RA. The common MTHFR C677T and A1298C variants are not associated with the risk of non-syndromic cleft lip/palate in northern Venezuela. Journal of genetics and genomics. J Genet Ge-nomics 2009 May;36(5):283-8. ##Shotelersuk V, Ittiwut C, Siriwan P, Angspatt A. Mater-nal 677CT/1298AC genotype of the MTHFR gene as a risk factor for cleft lip. J Med Genet 2003;40(5):e64.##Pezzetti F, Martinelli M, Scapoli L, Carinci F, Palmieri A, Marchesini J, et al. Maternal MTHFR variant forms in-crease the risk in offspring of isolated nonsyndromic cleft lip with or without cleft palate. Hum Mutat 2004; 24(1):104-5. ##de Araujo TK, Secolin R, Félix TM, de Souza LT, Fontes MÍ, Monlleó IL, et al. A multicentric association study be-tween 39 genes and nonsyndromic cleft lip and palate in a Brazilian population. J Craniomaxillofac Surg 2016; 44(1):16-20. ##Hu YY, Qin CQ, Deng MH, Niu YM, Long X. Associa-tion between BMP4 rs17563 polymorphism and NSCL/P risk: a meta-analysis. Dis Markers 2015;2015:763090. ##Chen Q, Wang H, Hetmanski JB, Zhang T, Ruczinski I, Schwender H, et al. BMP4 was associated with NSCL/P in an Asian population. PLoS One 2012;7(4):e35347.##Holder SE, Vintiner GM, Farren B, Malcolm S, Winter RM. Confirmation of an association between RFLPs at the transforming growth factor-alpha locus and non-syndromic cleft lip and palate. J Med Genet 1992;29(6): 390-2. ##Tanabe A, Taketani S, Endo-Ichikawa Y, Tokunaga R, Ogawa Y, Hiramoto M. Analysis of the candidate genes responsible for non-syndromic cleft lip and palate in Japa-nese people. Clin Sci (Lond) 2000;99(2):105-11. ##Stoll C, Qian JF, Feingold J, Sauvage P, May E. Genetic variation in transforming growth factor alpha: possible as-sociation of BamHI polymorphism with bilateral sporadic cleft lip and palate. Human Genet 1993;92(1):81-2. ##Lidral AC, Murray JC, Buetow KH, Basart AM, Schearer H, Shiang R, et al. Studies of the candidate genes TGFB2, MSX1, TGFA, and TGFB3 in the etiology of cleft lip and palate in the Philippines. Cleft Palate Cra-niofac J 1997;34(1):1-6.##Jugessur A, Rahimov F, Lie RT, Wilcox AJ, Gjessing HK, Nilsen RM,et al. Genetic variants in IRF6 and the risk of facial clefts: single-marker and haplotype-based analyses in a population-based case-control study of facial clefts in Norway. Genet Epidemiol 2008;32(5):413-24. ##Huang Y, Wu J, Ma J, Beaty TH, Sull JW, Zhu L, et al. Association between IRF6 SNPs and oral clefts in West China. J Dent Res 2009;88(8):715-8. ##Gurramkonda VB, Syed AH, Murthy J, Lakkakula B. IRF6 rs2235375 single nucleotide polymorphism is asso-ciated with isolated non-syndromic cleft palate but not with cleft lip with or without palate in South Indian popu-lation. Braz J Otorhinolaryngol 2018;84(4):473-477.##

Association of rs2013162 and rs2235375 Polymorphisms in IRF6 Gene with Susceptibility to Non-Syndromic Cleft Lip and Palate 2 2 Background: Non-syndromic cleft lip occurs by the interaction of environmental and genetic factors. The purpose of the current study was to analyze the association of Single Nucleotide Polymorphisms (SNPs) in IRF6 and NSCL/P in an Iranian population. Methods: A group of 105 children with NSCL/P and 185 normal controls were included in the current study. Genotyping of IRF6 rs2013162 and rs2235375 was performed by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method. Results: A substantial association of AA and CA genotypes in rs2013162 with the risk of NSCL/P (AA vs. CC; OR=2.36; 95%CI [1.05-5.29], p=0.004; and CA vs. CC; OR=0.47; 95%CI [0.28-0.79], p=0.018) was found. However, there were no important associations between A allele and risk of NSCL/P (p=0.980). According to logistic regression analysis results, subjects with GG genotype and G allele in rs2235375 polymorphism had increased risk of NSCL/P. Conclusion: The IRF6 polymorphisms are associated with the susceptibility to NSCL/P in Iranian population. 181 185 Masoumeh Soleymani Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Asghar Ebadifar Dentofacial Deformities Research Center, Research Institute of Dental Sciences, Department of Orthodontics, Faculty of Dentistry, Shahid Behehsti University of Medical Sciences Iran Maryam Khosravi Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Emran Esmaeilzadeh Fetal Health Research Center, Hope Generation Foundation Iran Hamid Reza Khorram Khorshid Genetics Research Center, University of Social Welfare and Rehabilitation SciencesPersonalized Medicine and Genometabolomics Research Center, Hope Generation Foundation IranIran IRF6Non-syndromic cleft lip and palateOrofacial cleftsPolymorphism 60502.pdf Mossey PA, Little J, Munger RG, Dixon MJ, Shaw WC. Cleft lip and palate. The Lancet 2009;374(9703):1773-85. ##Aldhorae KA, Böhmer AC, Ludwig KU, Esmail AHA, Al‐Hebshi NN, Lippke B, et al. Nonsyndromic cleft lip with or without cleft palate in arab populations: Genetic analysis of 15 risk loci in a novel case–control sample recruited in Yemen. Birth Defects Res A Clin Mol Teratol 2014;100(4):307-13. ##Yang J, Yu X, Zhu G, Wang R, Lou S, Zhu W, et al. Integrating GWAS and eQTL to predict genes and pathways for non‐syndromic cleft lip with or without palate. Oral Dis 2021;27(7):1747-54. ##Hassani M, Dehani M, Rafie MZ, Esmaeilzadeh E, Davar S, Pakzad B, et al. Investigation of rs531564 polymorphism in the primary microRNA-124 gene in patients with systemic lupus erythematosus and rheu-matoid arthritis: association with disease susceptibility and clinical characteristics. Iran J Allergy Asthma Im-munol 2021 Jun 6;20(3):303-13. ##Ishorst N, Francheschelli P, Böhmer AC, Khan MFJ, Heilmann-Heimbach S, Fricker N, et al. Nonsyndromic cleft palate: an association study at GWAS candidate loci in a multiethnic sample. Birth Defects Res 2018;110 (10):871-82. ##Leslie EJ, Marazita ML, editors. Genetics of cleft lip and cleft palate. Am J Med Genet C Semin Med Genet 2013; 163C(4):246-58. ##Mukhopadhyay N, Feingold E, Moreno-Uribe L, Wehby G, Valencia-Ramirez LC, Muñeton CPR, et al. Genome-wide association study of non-syndromic orofacial clefts in a multiethnic sample of families and controls identifies novel regions. Front Cell Dev Biol 2021;9:621482. ##Wattanawong K, Rattanasiri S, McEvoy M, Attia J, Thak-kinstian A. Association between IRF6 and 8q24 polymor-phisms and nonsyndromic cleft lip with or without cleft palate: Systematic review and meta‐analysis. Birth Defects Res A Clin Mol Teratol 2016;106(9):773-88. ##Martinelli M, Palmieri A, Carinci F, Scapoli L. Non-syndromic cleft palate: an overview on human genetic and environmental risk factors. 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Oral Dis 2019;25(1):223-33. ##Zucchero TM, Cooper ME, Maher BS, Daack-Hirsch S, Nepomuceno B, Ribeiro L, et al. Interferon regulatory factor 6 (IRF6) gene variants and the risk of isolated cleft lip or palate. New Engl J Med 2004;351(8):769-80. ##Huang Y, Wu J, Ma J, Beaty T, Sull J, Zhu L, et al. Association between IRF6 SNPs and oral clefts in West China. J Dent Res 2009;88(8):715-8. ##Ibarra-Arce A, García-Álvarez M, Cortés-González D, de Zarate-Alarcón GO, Flores-Peña L, Sánchez-Camacho S, et al. IRF6 polymorphisms in Mexican patients with non-syndromic cleft lip. Meta Gene 2015;4:8-16. ##Gurramkonda VB, Syed AH, Murthy J, Lakkakula BV. IRF6 rs2235375 single nucleotide polymorphism is associated with isolated non-syndromic cleft palate but not with cleft lip with or without palate in South Indian population. Braz J Otorhinolaryngol 2018;84:473-7. ##Suazo J, Recabarren AS, Marín NR, Blanco R. Associ-ation between IRF6 variants and nonsyndromic cleft lip with or without cleft palate in Chile. Reprod Sci 2020; 27(10):1857-62. ##Xu W, Han W, Lu Y, Feng W, Dai M. Association of single-nucleotide polymorphisms, rs2235371 and rs-2013162, in the IRF6 gene with non-syndromic cleft palate in northeast China. Genet Mol Res 2016;15(3). ##Lu Y, Liu Q, Xu W, Li Z, Jiang M, Li X, et al. TGFA and IRF6 contribute to the risk of nonsyndromic cleft lip with or without cleft palate in northeast China. PLoS One 2013;8(8):e70754. ##Velázquez-Aragón JA, Angel AG-d, Alcántara-Ortigoza MA, Reyna-Fabián ME, Estandia-Ortega B. Screening of IRF6 variants in patients subjected to genetic association studies for nonsyndromic cleft lip/palate. Cleft Palate-Craniofac J 2021;58(9):1128-34. ##