Avicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001Sortilin as a Novel Diagnostic and Therapeutic Biomarker in Chronic Lymphocytic Leukemia270276249728ENLiaFarahiMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IranFatemehGhaemimaneshMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IranSaeidehMilaniMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IranSeyed MohsenRazaviClinic of Hematology and Oncology, Firoozgar Hospital, Faculty of Medicine, Iran University of Medical Sciences, Tehran, IranMohammad MehdiAkhondiMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IranHodjattallahRabbaniMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IranJournal Article20180815Background: The overexpression of sortilin/neurotensin receptor 3 has previously been reported in various human solid tumors but not in hematological malignancies. Here, we report the overexpression of sortilin in leukemic cells from patients with Chronic Lymphocytic Leukemia (CLL). Methods: Flow cytometry was used to compare the expression of sortilin in CLL pa-tients (n=52) and healthy individuals (n=26). Also, in vitro apoptosis induction was assessed in CLL Peripheral Blood Mononuclear Cell (PBMCs) following directly targeting of sortilin. Results: The results showed a significant expression of sortilin on the surface of CLL PBMCs (range from 2.2 to 71.5%) in comparison to healthy individuals (range from 0.03 to 7.4%) (p≤0.0001). The optimal cut-off value of sortilin expression was deter-mined at 7.2% with high sensitivity and specificity. Treatment of leukemic cells with anti-sortilin antibody could induce apoptosis without any effect on normal cells. Conclusion: Apoptosis induction in CLL cells together with a significant correlation between the expression of sortilin and CD23 represent a possible functional role of sortilin in leukemogenesis of CLL cells. Therefore, sortilin might be considered as a promising novel biomarker in diagnosis, monitoring, and therapy of patients with CLL.https://www.ajmb.org/article_249728_97cb3d24859e4b8a69c1d0a0dd3b9ec6.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001Design and Fabrication of a Novel Microfluidic System for Enrichment of Circulating Tumor Cells with the Assistance of Computer Simulations277284249729ENDinaDorrigivDepartment of Chemical Engineering, Sharif University of Technology, Tehran, IranManouchehrVossoughiDepartment of Chemical Engineering, Sharif University of Technology, Tehran, IranIranAlemzadehDepartment of Chemical Engineering, Sharif University of Technology, Tehran, IranJournal Article20181020Background: Cancer is the first cause of death in developed countries. The heteroge-neous nature of cancer requires patient-specified treatment plans. One reliable ap-proach is collecting Circulating Tumour Cells (CTCs) and using them for prognosis and drug response assessment purposes. CTCs are rare and their separation from normal cell requires high-accuracy methods. Methods: A microfluidic cell capture device to separate CTCs from peripheral blood is presented in this study. The CTC separation device applies hydrodynamic forces to categorize cells according to their sizes. The proposed device is designed and evaluated by numerical simulations and validated experimentally. The simulation modified design was fabricated by soft lithography which allows prototyping the device in a few hours. For experimental setup two solutions: 1) fixed cells spiked in Phosphate Buffered Saline (PBS), and 2) fixed cells in blood were used. The CTC separation device was validated by tracking the flow and separation of cancer cell lines in the solutions. Results: It is demonstrated that the setup is capable of CTC enrichment up to 50 times. Conclusion: The presented CTC enrichment method reduces costs by eliminating the use of antibodies. The high-throughput method has the potential to be used in pre-clinical studies of cancer.https://www.ajmb.org/article_249729_011d0145b6ca8c123aa0cf3470bb8bb0.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001Anti-Influenza Virus Activity and Phenolic Content of Pomegranate (Punica granatum L.) Peel Extract and Fractions285291249730ENMohammad-TaghiMoradiMedical Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran0000-0002-3103-3740AliKarimiCellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, IranMehrdadShahraniCellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, IranLeilaHashemiClinical Biochemistry Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, IranMohammad-salehGhaffari-gooshehCellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, IranJournal Article20180610Background: Influenza virus, associated with high level of morbidity and mortality, has been recently considered a public health concern while the choices for the control and treatment of the disease are limited. The present study was conducted to evaluate activity of pomegranate peel extract and its fractions against Influenza A virus in vitro. Methods: In this research, ethyl alcohol extract of pomegranate peel was prepared and subjected to fractionation with different polarities. The potential in vitro anti-influenza A virus activity of the extract and fractions was assessed using Cytopathic Effect (CPE) reduction assay, Hemagglutinin Assay (HA), and 50% Tissue Culture In-fectious Doses (TCID50) method in Madin-Darby Canine Kidney (MDCK) cells. Results: The crude pomegranate peel extract and its n-butanol and ethyl acetate fractions had the highest inhibitory effect against influenza A virus with IC50 value of 6.45, 6.07 and 5.6 μg/ml in MDCK cells, respectively. Our results also showed that, the production of virus was significantly reduced upon treatment with crude extract, n-butanol and ethyl acetate fractions in a dose-dependent manner (p<0.05). Conclusion: Based on our results, the ethyl alcohol extract and its polar fractions of pomegranate peel can inhibit influenza A virus replication in vitro. Therefore, further characterization of its active ingredients and the mechanism of action should be carried out.https://www.ajmb.org/article_249730_e5410c077c88e310d472efe79aa3ff66.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001G2 Dendrimer as a Carrier Can Enhance Immune Responses Against HCV-NS3 Protein in BALB/c Mice292298249731ENFooziehJavadiDepartment of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran; Depratment of Biology, Science and Research Branch, Islamic Azad University, Tehran, IranPoonehRahimiDepartment of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, IranMohammad HossienModarresiDepartment of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, IranAzamBolhassaniDepartment of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, IranMehdiShafiee-ArdestaniDepartment of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, IranSeyed MehdiSadatDepartment of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, IranJournal Article20181119Background: Hepatitis C virus (HCV) infection is a major issue of public health. It seems of paramount importance to find an effective vaccine against HCV infection. The best vaccine candidate should induce robust cellular responses. The aim of the current study was to evaluate immunogenicity effects of novel conjugated dendrimer G2 with the recombinant NS3 antigen as a vaccine candidate for eliciting Th1-oriented cellular responses. Methods: Female BALB/c mice were immunized with different regimes especially with NS3 conjugated with G2 dendrimer. The humoral responses (Total IgG and IgG isotyping) and cellular responses (Ex vivo IFN-γ and IL-4 ELISpot assays, in vitro CTL assay and proliferation) were evaluated and compared in immunized mice. Results: The results indicated that induced specific total IgG in all mice groups im-munized with rNS3 formulated with different adjuvants and IgG2a subclass was the predominant isotype in rNS3-G2 (p≤0.05). For preliminary evaluation of cellular re-sponse, ex vivo ELISpot assay has shown that the higher frequency of IFN-γ producing cells was in groups immunized with rNS3+M720 and rNS3-G2 (p= 0.0012) than control groups. Finally, the rNS3-specific CTLs activity showed the highest percentage of specific lysis (LDH release) of the target cells in rNS3-G2 and rNS3+M720 groups. Conclusion: In the present study, as our knowledge, this is first time that the immunogenicity of nanodendrimer G2 as a biocompatible adjuvant with the HCV-NS3 antigen was evaluated. The results showed high capability of the regimen to induce strong Th1-orinted cellular response in mice model, indicating the dendrimer G2 as a novel adjuvant candidate for HCV vaccine studies.https://www.ajmb.org/article_249731_f44d49d5c7195b2acef61ce3288e3f2c.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001A Metabolomic Study to Identify Potential Tissue Biomarkers for Indomethacin-Induced Gastric Ulcer in Rats299307249732ENReyhanehFarrokhi-YektaProteomics Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IranNasrinAmiri-DashatanProteomics Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Student Research Committee, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, IranMehdiKoushkiDepartment of Clinical Biochemistry, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, IranMasoomehDadpayDepartment of Pathology, AJA University of Medical Sciences, Tehran, IranFatemehGoshadrouDepartment of Basic Sciences, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, IranJournal Article20180707Background: Gastric Ulcer (GU) is the most prevalent gastrointestinal disorder in-duced by various factors and Non-Steroid Anti-Inflammatory Drugs (NSAIDs) as one of the most common reasons. Due to the absence of appropriate molecular markers for GU, the aim of this study was to utilize a metabolomics approach in order to find potential metabolite markers for the disease. Methods: Stomach tissue samples from indomethacin-treated rats and normal con-trols were used to perform a 1H-NMR metabolomics study. The altered metabolites were identified using random forest multivariate analysis. Results: ROC curves showed that the random forest model had a good predictive performance with AUC of 1 for the test and 0.708 for the training sets. Seventeen differentially expressed metabolites were found between GU and normal tissue sample. These metabolites included trimethylamine, betaine, carnitine, methionine, acetylcho line, choline, N,N-Dimethylglycine, cis-aconitate, tryptophan, spermidine, acetylcar-nitine, creatinine, pantothenate, taurine, isoleucine, glucose and kynurenine. Conclusion: The results of the study demonstrated that metabolomics approach could serve as a viable method to find potential markers for GU. Surely, further studies are needed for the validation of the results.https://www.ajmb.org/article_249732_9f7672bb4ec757bdcc2d939b2fe9645b.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001Menstrual Blood Stem Cell Transplantation in Mice Model of Acute Liver Failure: Does Gender of Recipient Affect the Outcome?308316249733ENMinaFathi-KazerooniDepartment of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, IranGholamrezaTavoosidanaDepartment of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, IranJournal Article20180813Background: There exists a dramatic rise in liver failure and numerous patients un-dergo liver transplant for life-saving reasons annually. Introducing alternatives to allograft transplantation is necessary due to present limitations. Recently, a noninvasive stem cell population from Menstrual blood-derived Stem Cells (MenSCs) has been identified. There is an increasing interest in the application of MenSCs in tissue engineering; however, the fact that these gender-specific stem cells are safe for use in male sex is still not well defined. Methods: In this research, a model of acute liver failure was created in male and fe-male immunocompetent Balb-C mice through intraperitoneal injection of Carbon tetrachloride (CCl4) and MenSCs were transplanted intravenously 48 hrs after induc-tion of liver injury to evaluate their therapeutic potential. All mice were sacrificed on days 1, 7, and 30 post-transplantation to examine biochemical and molecular markers and pathological appearances. Results: Results showed the liver engraftment of MenSCs by immunofluorescence staining using anti-human mitochondrial antibody in both male and female treated groups. The restoration of serum markers of liver injury, aspartate aminotransferase and alanine aminotransferase, as well as expression levels of liver-specific genes, tyrosine aminotransferase and cholesterol 7 alpha-hydroxylase, were more significant in the female treated group compared with the male treated group on day 7 (p<0.05); however, after 30 days, there were no significant differences. Furthermore, hematoxylin and eosin and periodic acid-Schiff staining of liver sections demonstrated the considerable liver regeneration post cell therapy in both groups. Notably, data has shown that MenSCs could engraft into injured liver tissues and result in the same effect in the regeneration of liver function in both genders. Conclusion: Results of this study introduce MenSCs therapy as an attractive alterna-tive approach for liver repairing and regeneration which has no gender constraints.https://www.ajmb.org/article_249733_25955c055bff652b1412fce18a42af4a.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001Effect of Sodium Butyrate on LHX1 mRNA Expression as a Transcription Factor of HDAC8 in Human Colorectal Cancer Cell Lines317324249734ENMahsaGhiaghiDepartment of Genetics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, IranFloraForouzeshDepartment of Genetics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran0000-0002-2747-3479HamzehRahimiDepartment of Molecular Medicine, Pasteur Institute of Iran, Tehran, IranJournal Article20180926Background: LHX1 is an important transcription factor for the HDAC8 gene. The aim of this study was to investigate the effect of Sodium Butyrate (SB), as a histone deacetylase inhibitor, on the expression of LHX1 gene in colorectal cancer cell lines. Methods: HT-29 and HCT-116 cell lines were treated with 6.25 to 200 mM concentrations of SB at 24, 48, and 72 hr. The cytotoxicity effect on cell viability was evaluated by MTT assay. The 50% Inhibiting Concentration (IC50) was determined graphically. Quantitative real-time PCR was performed to investigate the LHX1 mRNA expression level. Results: Our study revealed that SB inhibited the proliferation of these cell lines in a concentration and time-dependent manner. The IC50 values for HT-29 cell line were 65, 18.6, and 9.2 mM after 24, 48, and 72 hr of treatment, respectively. The IC50 values for HCT-116 cell line were 35.5, 9.6, and 10 mM after 24, 48, and 72 hr of treatment, respectively. Furthermore, real-time PCR findings demonstrated that the LHX1 mRNA expression in treated HT-29 cell line significantly increased in comparison with untreated cells (p<0.05). However, in treated HCT-116 cell line, SB led to a significant decrease in the level of LHX1 mRNA (p<0.05), as compared to untreated cells. Conclusion: In this study, different effects of SB on LHX1 mRNA expression level were revealed in two distinct human colorectal cancer cell lines.https://www.ajmb.org/article_249734_499b3c9c6c8950b55783165ec6472fc7.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001Interleukin 10 and Transforming Growth Factor Beta Polymorphisms as Risk Factors for Kawasaki Disease: A Case-Control Study and Meta-Analysis325333249735ENFarzanehRahmaniResearch Center for Immunodeficiencies, Children’s Medical Center, Tehran University of Medical Sciences, Tehran, Iran; NeuroImaging Network (NIN), Universal Scientific Education and Research Network (USERN), Tehran, IranVahidZiaeePediatric Rheumatology Research Group, Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran; Department of Pediatrics, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of MRahelehAssariPediatric Rheumatology Research Group, Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran; Department of Pediatrics, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of MMaryamSadrMolecular Immunology Research Center, School of Medicine, Tehran University of Medical Sciences, Tehran, IranArezouRezaeiResearch Center for Immunodeficiencies, Children’s Medical Center, Tehran University of Medical Sciences, Tehran, IranZeinabSadrMolecular Immunology Research Center, School of Medicine, Tehran University of Medical Sciences, Tehran, IranSeyed RezaRaeeskaramiDepartment of Pediatrics, Vali-e-Asr Hospital, Tehran University of Medical Sciences, Tehran, IranMohammad HassanMoradinejadDepartment of Pediatrics, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of Medical Sciences, Tehran, IranYahyaAghighiDepartment of Pediatrics, Imam Khomeini Hospital, Tehran University of Medical Sciences, Tehran, IranNimaRezaeiResearch Center for Immunodeficiencies, Children’s Medical Center, Tehran University of Medical Sciences, Tehran, Iran; Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran; Network oJournal Article20180825Background: Alteration in serum expression of Transforming Growth Factor-beta (TGF-β) and IL-10 have been suggested to play a role in the pathogenesis of Kawa-saki Disease (KD). Inconsistent reports exist on the association of IL-10 polymorphisms with KD susceptibility and Coronary Artery Aneurysms (CAA). Methods: A number of 110 paediatric patients with KD and 140 healthy individuals were recruited to investigate the frequency of Single Nucleotide Polymorphisms (SNPs) of TGF-β C/T at codon 10 (rs1982073), C/G at codon 25 (rs1800471) and IL-10 A/G at -1082 (rs1800896), C/T at -819 (rs1800871) and A/C at -592 (rs1800872) and their respective genotype and haplotypes. A comprehensive search was performed in MEDLINE and SCOPUS using the keywords of interleukin 10, transforming growth factor beta, and Kawasaki disease. Moreover, previous studies investigating the TGF-β and IL-10 polymorphisms in KD were evaluated. Review Manager Version 5.1 Software was used to perform meta-analysis. Results: There was no significant association between allelic or genotypic variants in the mentioned polymorphisms in TGF-β or IL-10 with KD or CAA. The only significant haplotypic variant was TC variant at codon 10, and 25 of TGF-β polymorphisms were associated with higher risk of KD. Meta-analysis of a total number of 770 patients vs. 1471 healthy controls showed no difference in the frequency of any of the IL-10 genetic variants in KD patients, regardless of the presence of CAA. Conclusion: Polymorphisms of TGF-β or IL-10 are not associated with additional risk for KD in Iranian population. IL-10 polymorphisms at -1082, -819 and -592 positions are not associated with KD, nor do they predict coronary artery aneurysm formation.https://www.ajmb.org/article_249735_c71d3b7bc8b31436b42a1cfa02a4cc2a.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001Integrity and Quantity Evaluation of Plasma Cell-Free DNA in Triple Negative Breast Cancer334338249736ENMahdiehSalimiDepartment of Medical Genetics, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, IranSomayehSedaghati-BurkhaniDepartment of Medical Genetics, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, IranJournal Article20180815Background: Triple-Negative Breast Cancer (TNBC) is a subtype of breast cancer that lacks expression of the estrogen and progesterone receptor and does not overexpress human epidermal growth factor 2 receptor protein. TNBC is associated with special characteristics, including aggressiveness, poor prognosis, and treatment response. Non-invasive blood-based molecular markers such as cell-free DNA (cfDNA) variables have been shown to be putative markers in breast cancer prognosis. Methods: The cfDNA quantity and integrity were assessed in a case-control study of 96 breast cancer patients including 46 triple negative and 50 non-triple negative compared with 50 unaffected controls. A quantitative real-time PCR approach based on the quantification of two amplicons of the β-actin gene with different lengths (99 and 394 bp) was used to evaluate the integrity index 394/99. Results: Both cfDNA integrity index and quality were significantly elevated in breast cancer patients but integrity index can be considered as the more reliable diagnostic marker. The statistically significant increase of cfDNA quantity and integrity was ob-served in TNBC patients, somehow associated with nodal metastasis (p<0.001). Conclusion: Elevated cfDNA concentration and integrity index in breast cancer pa-tients compared with normal control and significant difference observed between TNBC and non-TNBC may be considered as a possible effective non-invasive diagnostic and prognostic molecular marker in breast cancer.https://www.ajmb.org/article_249736_bd986cafdf5138cf05ce9814685b48be.pdfAvicenna Research InstituteAvicenna Journal of Medical Biotechnology2008-283511420191001Importance of Fluctuating Amino Acid Residues in Folding and Binding of Proteins339344249737ENRenganathanSenthilDepartment of Bioinformatics, Faculty of Biosciences, The Marudupandiyar Institutions, Thanjavur-613403, Tamil Nadu, India0000-0002-8451-9832SingaraveluUshaDepartment of Bioinformatics, Bharathiar University, Coimbatore-641046, Tamil Nadu, IndiaKondaSaravananCentre of Advanced Study in Crystallography and Biophysics, University of Madras, Guindy Campus, Chennai 600025, Tamilnadu, IndiaJournal Article20180728Background: Conformational flexibility of proteins remains as one of the major events in protein-protein/DNA/ligand/small molecule binding to achieve its biological function in the cell. The availability of high-resolution structures of protein complexes is a valuable resource for researchers to understand the mechanisms behind such interactions and it is found that the flexibility of amino acid residues at binding sites is crucial for many important functions in the cell. Methods: In this article, our statistical method (PreFRP) developed based on fluctuating amino acid residues and various amino acid indices related to flexibility/rigidity were used to study the importance of fluctuating amino acid residues in thermonuc-leases from pathogenic bacteria, cell penetrating peptides and intrinsically disordered proteins responsible for many neural disorders. Results: The results from our analysis reveal the importance of fluctuating amino acid residues in folding and binding of proteins. The role of moderate and high fluctuating residues in themonucleases, cell penetrating peptide and disordered regions are discussed in detail. Conclusion: Therefore, our analysis will help in understanding the importance of fluc-tuating amino acid residues in proteins which undergo a conformation change phenomenon.https://www.ajmb.org/article_249737_a4c5e5a4c21c59eccc30148ee19f05fc.pdf