Cellular and Molecular Neurobiology (02724340)44(1)
The global public health addiction crisis has been stark, with over 932,400 deaths in the USA and Canada from opioid overdose since 1999–2020, surpassing the mortality rates at the top of the HIV/AIDS epidemic. Both nations exhibit opioid consumption rates significantly above the norm for developed countries. Analgesic type of opioids present both therapeutic benefits and substantial health risks, necessitating balanced drug regulation, careful prescribing, and dedicated opioid stewardship. The role of the cytochrome P450 2D6 (CYP2D6) system (Enzymatic functions) in metabolizing opioids highlights the potential of genotype-guided analgesia. By integrating Pharmacogenomics (PGx), this approach aims to optimize pain management, enhance safety, and reduce addiction risks. This understanding prompted the utilization of multifactor dimensionality reduction (MDR) to explore a range of phenotypes including PGx and gene–gene interactions (GGI) in a healthy cohort, thereby personalizing pain management strategies. The study sampled 100 unrelated healthy Western Iranians and 100 individuals from the 1000 Genome Project. Pre-testing involved searching for PGx annotations (variants associated with drug-gene-diseases) related to pain sensitivity and inflammation using the PharmGKB database, which identified 128 relevant genes. A questionnaire helped select 100 participants who had never used potent opioids but also other psychoactive agents (e.g., nicotine, amphetamines, etc.) and disease-related drugs. Whole-exome sequencing (WES) was then employed to analyze these genes in an Iranian cohort. Further analyses included MDR for identifying synergistic gene annotations and GGI for exploring complex gene interactions through the Visualization of Statistical Epistasis Networks (ViSEN). The study identified a Pain, Anti-Inflammatory, and Immunomodulating agents (PAIma) panel from the 128 genes, resulting in 55,590 annotations across 21 curated pathways. After filtering, 54 significant structural or regulatory variants were identified. This research also highlighted novel gene relationships involving the CYP3A5 gene, hsa-miR-355-5p, Paliperidone, and CYP2D6, which warrant further investigation. This study offers a novel pharmacogenetic framework that could potentially transform opioid prescribing practices to mitigate misuse and enhance personalized pain management. Further validation of these findings from multi countries and ethnic groups could guide clinicians in implementing DNA-based opioid prescribing, aligning treatment more closely with individual genetic profiles. Graphical abstract: (Figure presented.) © The Author(s) 2024.
Frontiers in Neurology (16642295)14
Introduction: Multiple sclerosis (MS), a non-contagious and chronic disease of the central nervous system, is an unpredictable and indirectly inherited disease affecting different people in different ways. Using Omics platforms genomics, transcriptomics, proteomics, epigenomics, interactomics, and metabolomics database, it is now possible to construct sound systems biology models to extract full knowledge of the MS and recognize the pathway to uncover the personalized therapeutic tools. Methods: In this study, we used several Bayesian Networks in order to find the transcriptional gene regulation networks that drive MS disease. We used a set of BN algorithms using the R add-on package bnlearn. The BN results underwent further downstream analysis and were validated using a wide range of Cytoscape algorithms, web based computational tools and qPCR amplification of blood samples from 56 MS patients and 44 healthy controls. The results were semantically integrated to improve understanding of the complex molecular architecture underlying MS, distinguishing distinct metabolic pathways and providing a valuable foundation for the discovery of involved genes and possibly new treatments. Results: Results show that the LASP1, TUBA1C, and S100A6 genes were most likely playing a biological role in MS development. Results from qPCR showed a significant increase (P < 0.05) in LASP1 and S100A6 gene expression levels in MS patients compared to that in controls. However, a significant down regulation of TUBA1C gene was observed in the same comparison. Conclusion: This study provides potential diagnostic and therapeutic biomarkers for enhanced understanding of gene regulation underlying MS. Copyright © 2023 Karimi, Motovali-Bashi and Ghaderi-Zefrehei.
Changizian, M.,
Nourisanami, F.,
Hajpoor, V.,
Parvaresh, M.,
Bahri, Z.,
Bashi naeini, M.M. Clinica Chimica Acta (00098981)536pp. 112-125
The significance of long non-coding RNAs (lncRNAs) in the development and progression of human cancers has attracted increasing attention in recent years of investigations. Having versatile interactions and diverse functions, lncRNAs can act as oncogenes or tumor-suppressors to actively regulate cell proliferation, survival, stemness, drug resistance, invasion and metastasis. LINC00467, an oncogenic member of long intergenic non-coding RNAs, is upregulated in numerous malignancies and its high expression is often related to poor clinicopathological features. LINC00467 facilitates the progression of cancer via sponging tumor-suppressive microRNAs, inhibiting cell death cascade, modulating cell cycle controllers, and regulating signalling pathways including AKT, STAT3, NF-κB and Wnt/β-catenin. A growing number of studies have revealed that LINC00467 may serve as a novel prognostic biomarker and its inhibitory targeting has a valuable therapeutic potential to suppress the malignant phenotypes of cancer cells. In the present review, we discuss the importance of LINC00467 and provide a comprehensive collection of its functions and molecular mechanisms in a variety of cancer types. © 2022
International Journal Of Reproductive Biomedicine (24764108)20(5)pp. 399-404
Background: Some previous human and animal studies have supported the idea that KDM3A down-regulation might be the main cause of male infertility, especially in non-obstructive azoospermia (NOA). The regulatory role of micro-RNAs (miRNA) has been investigated in the development of male infertility. Objective: The expression level of hsa-miR-30a-5p in azoospermia was evaluated to reveal its possible association with the etiology of male infertility. Materials and Methods: In this case-control study, 30 men with azoospermia (19 of whom had NOA) were selected as the case individuals, and 11 men with obstructive azoospermia (OA) were selected as control individuals. The best miRNA with the strongest ability to target the KDM3A gene was detected via comprehensive bioinformatics analysis. Reverse transcriptase quantitative polymerase chain reaction was used to assess the expression level of hsa-miR-30a-5p. After analyzing the data, the expression level of hsa-miR-30a-5p was compared between men with NOA and men with OA. Results: The findings supported the idea that hsa-miR-30a-5p is the miRNA with the best ability to target the KDM3A transcript. The expression analysis of hsa-miR-30a-5p indicated a significant overexpression (p = 0.04) in men with NOA compared to in men with OA. Conclusion: Hsa-miR-30a-5p was overexpressed in men with NOA compared to in control individuals. Hsa-miR-30a-5p could target the KDM3A transcript and may suppress its expression. © 2022, Research and Clinical Center for Infertitlity. All rights reserved.
Dehbashi, M.,
Hojati najafabadi, Z.,
Bashi naeini, M.M.,
Ganjalikhany, M.R.,
Cho, W.C.,
Shimosaka, A.,
Navabi, P.,
Ganjalikhani-hakemi, M. Frontiers in Oncology (2234943X)11
For many years, high-affinity subunit of IL-2 receptor (CD25) has been considered as a promising therapeutic target for different pathologic conditions like allograft rejection, autoimmunity, and cancers. Although CD25 is transiently expressed by newly-activated T cells, it is the hallmark of regulatory T (Treg) cells which are the most important immunosuppressive elements in tumor microenvironment. Thus, Tregs can be considered as a potential target for chimeric antigen receptor (CAR)-based therapeutic approaches. On the other hand, due to some profound adverse effects pertaining to the use of CAR T cells, CAR NK cells have caught researchers’ attention as a safer choice. Based on these, the aim of this study was to design and develop a CAR NK cell against CD25 as the most prominent biomarker of Tregs with the prospect of overcoming immune escape mechanism in solid and liquid cancers. In the current study, an anti-CD25 CAR was designed and evaluated by comprehensive in silico analyses. Then, using lentiviral transduction system, NK-92 cell line was engineered to express this anti-CD25 CAR construct. In vitro functional analyses of anti-CD25 CAR for its reactivity against CD25 antigen as well as for cytotoxicity and cytokine production assays against CD25 bearing Jurkat cell line were done. In silico analyses demonstrated that the anti-CD25 CAR transcript and scFv protein structures were stable and had proper interaction with the target. Also, in vitro analyses showed that the anti-CD25 CAR-engineered NK-92 cells were able to specifically detect and lyse target cells with an appropriate cytokine production and cytotoxic activity. To conclude, the results showed that this novel CAR NK cell is functional and warrant further investigations. © Copyright © 2021 Dehbashi, Hojati, Motovali-bashi, Ganjalikhany, Cho, Shimosaka, Navabi and Ganjalikhani-Hakemi.
Dehbashi, M.,
Hojati najafabadi, Z.,
Bashi naeini, M.M.,
Ganjalikhani-hakemi, M.,
Shimosaka, A.,
Cho, W.C. Biological Chemistry (14316730)402(2)pp. 167-178
Cancer recurrence presents a huge challenge in cancer patient management. Immune escape is a key mechanism of cancer progression and metastatic dissemination. CD25 is expressed in regulatory T (Treg) cells including tumor-infiltrating Treg cells (TI-Tregs). These cells specially activate and reinforce immune escape mechanism of cancers. The suppression of CD25/IL-2 interaction would be useful against Treg cells activation and ultimately immune escape of cancer. Here, software, web servers and databases were used, at which in silico designed small interfering RNAs (siRNAs), de novo designed peptides and virtual screened small molecules against CD25 were introduced for the prospect of eliminating cancer immune escape and obtaining successful treatment. We obtained siRNAs with low off-target effects. Further, small molecules based on the binding homology search in ligand and receptor similarity were introduced. Finally, the critical amino acids on CD25 were targeted by a de novo designed peptide with disulfide bond. Hence we introduced computational-based antagonists to lay a foundation for further in vitro and in vivo studies. © 2020 Walter de Gruyter GmbH, Berlin/Boston 2020.
Molecular Biology Research Communications (2322181X)10(2)pp. 45-53
Hemophilia A is an X-linked bleeding disorder that occurs due to the deficiency of Factor VIII (FVIII) protein clotting activity. The mutations in the F8 gene, which encodes FVIII coagulating protein have been widely reviewed. However, there is a wide range of criteria that in addition to F8 gene mutations, different molecular mechanisms may be associated with hemophilia A. Various functions of FVIII could be related to the hypothetical small non-coding RNAs, located within the F8 gene sequence. Therefore, miRNAs that can post-transcriptionally regulate gene expression might confer susceptibility to developing hemophilia A. Here, we have selected a bioinformatically predicted hairpin structure sequence in the first intron of the F8 gene that has the potential to produce a real miRNA (named put-miR1). We tried to experimentally detect the predicted miRNA via RT-PCR following its precursor overexpression in HEK 293 cell lines. Despite the accuracy of miRNA prediction, according to the reliable bioinformatics studies, we couldn’t confirm the existence of considered mature miRNA in transfected cells. We hope that through changing experimental conditions, designing new primers, or altering cell lines and expression vectors, the exogenous and endogenous expression of the predicted miRNA will be confirmed. © 2021. All Rights Reserved
Iranian Journal Of Biotechnology (23222921)19(2)pp. 64-70
Background: Hemophilia A is an X-linked bleeding disorder resulting in a deficiency of plasma clotting factor VIII and caused by mutations in the FVIII gene (F8 gene). MicroRNAs (miRNAs) in body fluids are promising biomarker candidates for Hemophilia A, due to their stability in body fluids and accessibility by non-or minimally-invasive procedures. Therefore; Advances in miRNA analysis methods resulted in a wide range of publications on miRNAs as putative biomarkers. Objective: Here we tried to scan the F8 gene region to predict a novel miRNA and identify it as a regulator of the F8 gene. Materials and Methods: To this aim, the ability to express novel miRNAs in F8 locus was assessed via reliable bioinformatics databases such as SSCprofiler, RNAfold, miREval, FOMmiR, MaturBayes, miRFIND, UCSC genome browser, Deep Sequencing, and miRBase. Results: Data analysis from the relevant databases offers one stem-loop structure that is predicted to express a novel miRNA. Conclusions: The diagnosis of Hemophilia A with the help of these types of biomarkers is a non-invasive procedure that has been demonstrated to have a significant role in the early diagnosis of the disease. Hopefully, the proposed candidate sequence will be confirmed in vitro and become a non-invasive biomarker in the near future. © 2021 The Author(s);.
Cell Journal (Yakhteh) (22285806)23(3)pp. 341-348
Objective: Hemophilia-A is a common genetic abnormality resulted from decreased or lack of factor VIII (FVIII) pro-coagulant protein function caused by mutations in the F8 gene. Majority of molecular studies consider screening of mutations and their relevant impacts on the quality and expression levels of FVIII. Interestingly, some of the functions in FVIII suggest a probable involvement of small non-coding RNAs embedded within the sequence of F8 gene. Therefore, microRNAs which are encoded within the F8 gene might have a role in hemophilia development. In this study, miRNAs production in the F8 gene was investigated by bioinformatics prediction and experimental validation. Materials and Methods: In this experimental study, bioinformatics tools have been utilized to seek the novel microRNAs inserted within human F8 gene. The ability to express new microRNAs in F8 locus was studied through reliable bioinformatics databases such as SSCProfiler, RNA fold, miREval, miR-FIND, UCSC genome browser and miRBase. Then, expression and processing of the predicted microRNAs were examined based on bioinformatics methods, in the HEK293 cell lines. Results: We are unable to confirm existence of the considered mature microRNAs in the transfected cells. Conclusion: We hope that through changing experimental conditions, designing new primers or altering cell lines as well as the expression of vectors, exogenous and endogenous expressions of the predicted miRNA will be confirmed. © 2021 Royan Institute (ACECR). All rights reserved.
Scientific Reports (20452322)11(1)
Non-enzymatic glycation of DNA and the associated effects are among pathogenic factors in diabetes mellitus. Natural polyphenols have anti-diabetic activity. Herein, the protective role of one of the phytochemicals, rosmarinic acid (RA), was evaluated in glycation (with fructose) of human DNA and expression of Akt genes in the hippocampus of diabetic rats. In-vitro studies using fluorescence, agarose gel electrophoresis, fluorescence microscopy, and thermal denaturation analyses revealed that glycation causes DNA damage and that RA inhibits it. In-vivo studies were performed by induction of diabetes in rats using streptozotocin. The diabetic rats were given RA daily through gavage feeding. The expression of Akt genes (inhibitors of apoptosis) in the hippocampus was evaluated using RT-qPCR. In diabetic rats, Akt1 and Akt3 were significantly down-regulated compared to the control group. Treating the diabetic rats with RA returned the expression of Akt1 and Akt3 relatively to the normal condition. Past studies have shown that diabetes induces apoptosis in the hippocampal neurons. Given that glycation changes the genes expression and causes cell death, apoptosis of the hippocampal neurons can be due to the glycation of DNA. The results also suggest that RA has reliable potency against the gross modification of DNA under hyperglycemic conditions. © 2021, The Author(s).
Dehbashi, M.,
Hojati najafabadi, Z.,
Bashi naeini, M.M.,
Cho, W.C.,
Shimosaka, A.,
Ganjalikhani-hakemi, M. Gene Reports (24520144)24
The regulatory T cells function in the immune homeostasis, these cells express high level of CD25. It is known that CD25 expression levels are various among different cancers. Thus, we intended to dissect systems biology of a lncRNA pertained to CD25 and CD25 protein interactors-targeting miRNAs. Using RNA-seq data, co-expression analyses of the lncRNA pertained to some cancers were performed. Our analysis was done for protein interactors of CD25 by STRING 11.0, ShinyGO v0.60 and KEGG web servers were used for enrichment and network analysis of CD25. TargetScan 7.2, miRTargetLink Human and mirDIP were applied for determining the CD25 and CD25 interactors-targeting miRNAs. To find the lncRNA-miRNA and lncRNA-protein interactions, starBase v3.0, LncBase Predicted v.2 and SFPEL-LPI were utilized, respectively. Also, using Co-LncRNA, the co-expressed lncRNA analysis and the relative signaling pathways in some cancers including bladder, breast, head and neck, kidney, liver, lung, prostate and thyroid cancers using RNA-seq data were achieved. OIP5-AS1 was shown to have the interaction with CD25 and CD25 protein interactors-targeting miRNAs. In addition, the co-expression of OIP5-AS1 in cancers and their signaling pathways was identified. In conclusion, OIP5-AS1 can effect on CD25 expression in all relative signaling pathways of these cancers. © 2021 Elsevier Inc.
Fazeli, S.,
Bashi naeini, M.M.,
Peymani, M.,
Hashemi, M.,
Etemadifar, M.,
Nasr-esfahani, M.H.,
Ghaedi, K. PLoS ONE (19326203)15(11 November)
Parkinson’s disease (PD) is diagnosed when motor symptoms emerges, which almost 70% of dopamine neurons are lost. Therefore, early diagnosis of PD is crucial to prevent the progress of disease. Blood-based biomarkers, which are minimally invasive, potentially used for diagnosis of PD, including miRNAs. The aim of this study was to assess whether SRRM2 and miR-27a/b-3p could act as early diagnostic biomarkers for PD. Total RNAs from PBMCs of 30 PD’s patients and 14 healthy age and gender matched subjects was extracted. The expression levels of respective genes were assessed. Data were presented applying a two-tailed unpaired t-test and one-way ANOVA. We observed significant downregulation of SRRM2 (p = 0.0002) and miR-27a-3p (p = 0.0001), and up-regulation of miR-27b-3p (p = 0.02) in PBMCs of Parkinson’s patients. Down-regulation of miR-27a-3p is associated with increasing disease severity, whereas the up-regulation of miR-27b-3p was observed mostly at HY-1 and disease duration between 3–5 years. There was a negative correlation between SRRM2 and miR-27b-3p expressions, and miR-27a-3p positively was correlated with miR-27b-3p. Based on functional enrichment analysis, SRRM2 and miR-27a/b-3p acted on common functional pathways. miR-27a/b-3p could potentially predict the progression and severity of PD. Although both miRs had no similarity on expression, a positive correlation between both miRs was identified, supporting their potential role as biomarkers in clinical PD stages. Of note that SRRM2 and miR-27a-3p were able to distinguish PD patients from healthy individuals. Functional analysis of the similarity between genes associated with SRRM2 and miR-27a/b-3p indicates common functional pathways and their dysfunction correlates with molecular etiopathology mechanisms of PD onset. © 2020 Fazeli et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Hemoglobin (03630269)44(1)pp. 27-30
β-Thalassemia intermedia (β-TI) is a clinical condition characterized by moderate, non transfusional anemia and hepatosplenomegaly. The main objective of this study was to determine the molecular basis of the clinical phenotype of β-TI in Iran. To elucidate the mild phenotype of many patients with β-TI, we screened for three prevalent β-globin gene mutations [IVS-II-1 (G>A) HBB: c.315+1G>A, IVS-I-110 (G>A) HBB: c.93-21G>A and IVS-I-5 (G>C) [HBB: c.92+5G>C], deletions on the α-globin genes, XmnI polymorphisms and restriction fragment length polymorphism (RFLP) haplotypes on the β-globin gene cluster in 50 β-TI patients. Fifty-eight percent of the patients (29 cases) were associated with the mentioned mutations. We showed that the HBB: c.315+1G>A mutation is linked to haplotype [+–+ +] (57.69%). This haplotype is in linkage disequilibrium with the XmnI polymorphism (NG_000007.3: g.42677C>T) and has been associated with increased expression of Hb F in β-TI patients. The XmnI polymorphism is defined in association with this prevalent mutation. Two patients had a single α-globin gene deletion [–α3.7 (rightward) deletion]. The main genetic factor in mild phenotype β-TI patients is the linkage of an XmnI polymorphism (NG_000007.3: g.42677C>T) with the HBB: c.315+1G>A (80.76%), which is associated with increased production of Hb F and coinheritance of haplotype [+–+ +] with β-TI, especially with the homozygous HBB: c.315+1G>A mutation. Molecular basis of β-TI could be explained by the involvement of different factors that tend to develop the disease phenotype. © 2020, © 2020 Informa UK Limited, trading as Taylor & Francis Group.
Fazeli, S.,
Bashi naeini, M.M.,
Peymani, M.,
Hashemi, M.,
Etemadifar, M.,
Nasr-esfahani, M.H.,
Ghaedi, K. PLoS ONE (19326203)15(12 December)
There is an error in affiliation 2 for author Maryam Peymani. The correct affiliation 2 is: Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran. © 2020 Public Library of Science. All rights reserved.
International Journal Of Molecular And Cellular Medicine (22519645)9(1)pp. 33-49
A major complication in treating hemophilia A is the development of neutralizing antibodies (inhibitors) against therapeutic administered factor VIII (FVIII), which occurs in approximately 20-30% of patients with severe disease. These inhibitors render FVIII replacement therapy ineffective and increase the morbidity and mortality risk. The currently accepted method to eradicate inhibitors is immune tolerance induction (ITI), and frequent intensive administration of FVIII until inhibitor titers drop. Current ITI protocols are extremely costly and not effective in all patients. During the last decade, many types of research have been accomplished to clarify the mechanisms that mediate immune tolerance induction. Novel experimental therapies including monoclonal antibodies, viral vector-mediated gene therapy, regulatory T cell induction using immunosuppressive drugs, and nanoparticle-based immune modulation show promising results in hemophilia A clinical trials. This review focuses on treatment options towards the anti-FVIII immune responses and current novel therapies in clinical trials. © 2020, Babol University of Medical Sciences.
International Journal Of Reproductive Biomedicine (24764108)18(11)pp. 961-968
Background: The role of KDM3A and its downstream genes in male fertility has been approved in animal models. Additionally, the expression shrinkage of KDM3A is significantly correlated with human azoospermia phenotype. Aberrant expression of micro-RNAs could mislead spermatogenesis and mostly lead to diverse phenotypes of male infertility. Objective: The aim of this study was to evaluate the expression level of hsa-miR-27a-3p in azoospermic men to reveal its possible association with infertility. Materials and Methods: This case-control study was conducted on 30 azoospermic men, of whom, 19 had non obstructive azoospermia (NOA) and 11 obstructive azoospermia (OA) according to the pathological examinations. Comprehensive bioinformatics investigations were performed securely and hsa-miR-27a-3p was selected afterward. Reverse Transcriptase-quantitative polymerase chain reaction (RT-qPCR) method was used and statistical analysis was performed to compare the expression level of hsa-miR-27a-3p in both OA and NOA individuals. Results: In silico analysis suggested hsa-miR-27a-3p, with its potential binding ability to target KDM3A transcripts. The expression analysis of candidate hsa-miR-27a-3p indicated its significant overexpression in NOA men. Conclusion: The hsa-miR-27a-3p was overexpressed in NOA men compared to OA-control individuals. As a consequence, the overexpressed micro-RNA could downregulate directly KDM3A and indirectly TNP1 and PRM1. Therefore, spermatogenesis could be misled and male infertility could be developed. © Norioun et al.
Analytica Chimica Acta (00032670)1092pp. 66-74
A sensitive and selective electrochemical method for simultaneous detection of two hemophilia A-related microRNAs (miR-1246 and miR-4521) was developed. This detection tactic was based on gold nanoparticles (AuNPs), heavy metals quantum dots-encapsulated metal-organic frameworks (QDs@ZIF-8), and catalytic hairpin assembly (CHA) for signal application. Primarily, hairpins H1 and H2 were hybridized with targets miR-1246 (T1) and miR-4521 (T2) for forming H1-T1 and H2-T2 duplex stranded DNAs (dsDNAs) that were able to open the hairpins H3 and H4 for the formation of H1–H3 and H2–H4 dsDNAs. Meanwhile, lots of H1–H3 and H2–H4 dsDNAs were created by releasing the target to take part in the next cycle for signal amplification. And then single stranded fragments of H1–H3 and H2–H4 dsDNAs were utilized for hybridizing the PbS@ZIF-8-S1 and CdS@ZIF-8-S2 in order to amplify the electrochemical signal. The diagnosis of corresponding target miRs using differential pulse voltammetry has been possible by releasing Pb (II) and Cd (II) ions from PbS@ZIF-8 and CdS@ZIF-8 tags by HCI leaching. In this context, encapsulation of heavy metals quantum dots (QDs) was done in zeolitic imidazolate framework-8 (ZIF-8) to form QDs@ZIF-8 muti-core-shell particles by in situ growth of ZIF-8 in the presence of QDs. Since the quantity of QDs tagged to each target miRs grows massively, being resulted from a huge number of QDs that encapsulated in each QDs@ZIF-8 label, the sensitivity of the biosensor using QDs@ZIF-8 particles as signal tags is about 15 times that of a biosensor using QDs as signal tags. Several conditions of determination like incubation time for labeling and capture probe, HCl leaching time, and reaction time of CHA were optimized. Under the optimized conditions, this assay allowed the detection of target miRs in the range of 1 fM to 1 μM with detection limits of 0.19 fM and 0.28 fM for miR-1246 and miR-4521 (S/N = 3). The biosensor can discriminate complementary, 1-base mismatched and non-complementary sequences quite well, according to the catalytic hairpin assembly. Furthermore, the biosensor was utilized efficiently for quick and direct analysis of microRNAs in human serum. Thus, this tactic presents an innovative platform for microRNAs expression profiling in biomedical research and clinical diagnosis. © 2019 Elsevier B.V.
Gene (18790038)687pp. 272-279
Purpose: MicroRNAs are involved in diverse biological processes and their dysregulation is a common event in various diseases including breast cancer. Breast cancer is a major threat to women's health. This study was designed to examine the expression levels of miR-9 and miR-34a in breast tumor tissue samples and plasma of breast cancer patients, compare their expression pattern between tissue samples and plasma samples of patients and analyze their relationship with tumor clinical features. Also, the potential of these miRNAs as diagnostic biomarkers for breast cancer was investigated. Materials and methods: The expression levels of miR-9, miR-34a and CDH1 were measured by real-time reverse transcription polymerase chain reaction and ΔΔct method. Data were analyzed using t-test and one-way ANOVA. The sensitivity and specificity of miRNAs were determined by receiver operating characteristic (ROC) curve. Results and discussion: The expression levels of miR-9 and miR-34a were significantly down-regulated in tumor tissues compared to healthy tissues (fold change = 0.26, p = 0.0051 for miR-9 and fold change = 0.55, p = 0.021 for miR-34a). While no significant difference was observed in the expression levels of miR-9 (p = 0.205) and miR-34a (p = 0.132) in plasma samples of patients compared to normal plasma. CDH1 expression in tumor tissue was not significantly different from normal tissue (p = 0.33). We found that expression level of miR-9 in patients with tumor size larger than 5 cm (p = 0.026) and expression level of miR-34a in patients with higher stage (lll & lV, p = 0.03) were significantly down-regulated. Also miR-34a expression level was positively correlated with patient's age (p = 0.03). Conclusion: According to the ROC curves, the area under the curve (AUC) of miR-9 in tissue was 0.71 (p = 0.009) with sensitivity 83.33% and specificity 70.37%. The AUC for miR-34a in tissue was 0.72 (p = 0.007) with sensitivity 72% and specificity 76%. Thus miR-9 and miR-34a have the capability for distinguishing tumor tissues from healthy tissues and the study of their expression levels in tissue may be used as a biomarker for the diagnosis of breast cancer patients from healthy women. © 2018
Biomolecular Concepts (1868503X)10(1)pp. 150-159
Typically, CD25 is expressed on the cellular surface of regulatory T (Treg) cells. These cells are significant in regulating the self-tolerance and also preventing the immune system from attacking a person's own tissues and cells. They promote the cancer progression by playing an important role in evading the immune system. Thus, the experimental procedures was aimed to clone and express human CD25 in HEK293 cell line, as the available cellular model, for the purpose of developing assays to facilitate and enhance the studies on an available CD25 positive cell. The secondary RNA structure of CD25 was evaluated by in silico analysis. Then, cDNA of human CD25 were synthesized from isolated total mRNA of cultured and stimulated PBMCs from blood donors. After cloning the cDNA of CD25 into a pcDNA3.1(+) plasmid, using the effective transfection of the recombinant pcDNA3.1(+) in HEK293, qRT-PCR and flow cytometry methods were used to quantitatively evaluate CD25 transcripts and protein level. There was a 4.8 fold increase in transcripts and a 76.2% increase in protein levels of CD25 when comparing the transfected and control cell lines. The genetically engineered HEK293 cell line expressing Treg cell surface marker of CD25 was introduced in this study for the first time. This cell line can be used to overcome the problematic issues for studying Treg cells including low population of Tregs in peripheral blood, low recovery methods for Treg isolation, time-consuming and non-cost benefit methods in the conditions of in vitro cell culture experiments for the studies focused on the binding of IL-2 to CD25. © 2019 Moein Dehbashi et al.
International Journal Of Hematology-Oncology And Stem Cell Research (17351243)13(2)pp. 61-67
Introduction: Beta-thalassemia is one of the most prevalent inherited blood diseases among Iranians. The aim of this study was to elucidate the chromosomal background of beta-thalassemia mutations in Esfahan province, Iran. Materials and Methods: In this study, we investigated three frequent mutations (c.315+1G>A, c.93-21G>A and c.92+5G>C in β-globin gene, the frequency of RFLP haplotypes, and LD between markers at β-globin gene cluster) in 150 beta-thalassemia patients and 50 healthy individuals. The molecular and population genetic investigations were performed on RFLP markers HindIII in the c.315+1G>A of Gã (HindIIIG) and Aã (HindIIIA) genes, AvaII in the c.315+1G>A of β-globin gene and BamHI 3' to the β-globin gene. All statistical analyses were performed using Power Marker software and SISA server. Results: Fifty percent of beta-thalasemia patients were associated with these mutations. Haplotype I was the most prevalent haplotype among beta-thalassemia patients (39.33%) and normal individuals (46%). The commonest c.315+1G>A mutation in our population was tightly linked with haplotype III (43.75%) and haplotype I (31.25%). The second prevalent mutation, c.92+5G>C, was 90%, 6.66%, and 3.33% in linkage disequilibrium with haplotypes I, VII, and III, respectively. The c.93-21G>A mutation indicated a strong association with haplotype I (80%). Conclusion: Our study participants like beta-thalassemia patients from Kermanshah province was found to possess a similar haplotype background for common mutations. The emergence of most prevalent mutations on chromosomes with different haplotypes can be explained by gene conversion and recombination. High linkage of a mutation with specific haplotype is consistent with the hypothesis that chromosomes carrying beta-thalassemia mutations experienced positive selection pressure, probably because of the protection against malaria experienced by beta-thalassemia carriers. © 2019, Tehran University of Medical Sciences (TUMS). All rights reserved.
Current Molecular Medicine (15665240)19(7)pp. 461-472
The demands for genotyping techniques with acceptable precision, accuracy, cost-effectiveness in high throughput formats made driving forces for continuous development of novel technologies. A wide range of mutation detection techniques based on polymerase chain reaction (PCR) have been introduced. The best alternatives were the isothermal amplification technologies that those did not require a thermal cycler. In this review, we aimed to describe the most known isothermal amplification techniques for SNP genotyping. © 2019 Bentham Science Publishers.
Mehraban, M.H.,
Mansourian, M.,
Ahrari, S.,
Hajiebrahimi, A.,
Odooli, S.,
Bashi naeini, M.M.,
Yousefi, R.,
Ghasemi, Y. Computational Biology and Chemistry (14769271)82pp. 25-36
The prevalence of diabetes mellitus has been incremented in the current century and the need for novel therapeutic compounds to treat this disease has been significantly increased. One of the most promising approaches is to inhibit intestinal alpha glucosidases. Based on our previous studies, four pyrimidine-fused heterocycles (PFH) were selected as they revealed satisfactory inhibitory action against mammalian α-glucosidase. The interaction of these compounds with both active domains of human maltase-glucoamylase (MGAM) and their effect on human Caco-2 cell line were investigated. The docking assessments suggested that binding properties of these ligands were almost similar to that of acarbose by establishing hydrogen bonds especially with Tyr1251 and Arg526 in both C-terminal and N-terminal MGAM, respectively. Also, these compounds indicated a stronger affinity for C-terminal of MGAM. L2 and L4 made tightly complexes with both terminals of MGAM which in turn revealed the importance of introducing pyrimidine scaffold and its hinge compartment. The results of molecular dynamics simulation analyses confirmed the docking data and showed deep penetration of L2 and L4 into the active site of MGAM. Based on cell cytotoxicity assessments, no significant cell death induction was observed. Hence, these functional MGAM inhibitors might be considered as new potential therapeutic compounds in treatment of diabetes and its complications. © 2019
Biochemical and Biophysical Research Communications (0006291X)519(1)pp. 192-197
Type II diabetes is a metabolic disease that has affected 460 million people around the globe and become a heavy burden on health care system. Diabetic patients suffer from hyperglycemia and hyperinsulinemia which can damage vital organs in body like heart, kidneys, eyes and nervous system. Different strategies have been introduced to control or lessen these diabetic complications in which one of the most promising approaches is the inhibition of intestinal sucrase-isomaltase (SI). Inhibition of this enzyme will block the release of glucose into bloodstream and lead to reduced postprandial hyperglycemia. MicroRNAs are small regulatory molecules that play critical roles in different cellular pathways and molecular mechanisms. It is proved that microRNAs have significant effects on cellular mechanisms involved in diabetes and can be used as biomarkers for diagnosis of this metabolic disease. Based on bioinformatics analysis miR-26a and miR-26b can interact with a conserved 3′-UTR region of SI mRNA which lead to a hypothesis that these miRs may have negative regulatory effect on this enzyme. In this study, we investigated the impact of high glucose conditions on expression of sucrase-isomaltase, miR-26a and miR-26b in caco-2 cell line. It is proved that in a simulated diabetic condition there is a reverse correlation between the expression pattern of these miRs and SI. QRT-PCR method was used to evaluate the expression of our target molecules. Interestingly, transfection of miR-26a and miR-26b in caco-2 cell line reduced the transcription of SI mRNA and decreased the sucrase and maltase activity of its active sites. To sum up, our results demonstrate the first evidence of the significant effect of miR-26a and miR-26b on SI expression and activity. We proved that these microRNAs may directly inhibit this enzyme and can be used as a new scaffold in search of finding novel treatments for type II diabetes. © 2019 Elsevier Inc.
The etiology of Multiple Sclerosis (MS) as a multifactorial neurodegenerative disease is still mostly unknown. Various “omic” approaches, including genome-wide association studies, transcriptome analysis, exome sequencing, and epigenome studies are considered to be helpful for better understanding of MS progression and pathophysiology as well as more accurate determination of different MS subtypes. In recent years, the importance of epigenetics in MS pathogenesis, as well as other autoimmune diseases, has been surprisingly well received. Epigenetic therapy, which includes drugs that have the ability to influence the reversible and dynamic epigenetic characteristics, is also identified as a promising therapeutic tool in MS. This chapter will provide a summary of recent studies considering potential roles of epigenetic alterations as biomarkers in MS. © 2018 Elsevier Inc. All rights reserved.
Molecular Biology Reports (03014851)45(4)pp. 413-417
Infertility occurs in 10–15% of couples worldwide and close to half of it is caused by male factors. One of the genes that can affect male infertility is CGA. Polymorphisms in CGA gene may affect gene expression, therefore affecting male infertility by disrupting the regulation of this gene. One of the polymorphisms is the substitution of T with A in the miR-1302 binding site in the 3′ untranslated region of the CGA gene. In this study, we explored this polymorphism in Isfahan population. In this case-control study, by the use of Tetra primer-ARMS–PCR technique, rs6631 has been investigated in 224 infertile men and 196 controls. Infertile men were recruited from Isfahan Fertility and Infertility Center. Analysis of genotype and allele frequencies indicated that the differences between case and control populations were significant for rs6631 because P = 0.00 which is above the threshold. We found a significant relationship between this polymorphism and male infertility. This study which performed for the first time in Iran suggests that polymorphism in CGA gene can affect male infertility. Also, this polymorphism has high heterozygosity, so it can be used for further studies in different populations. © 2018, Springer Science+Business Media B.V., part of Springer Nature.
Journal of Isfahan Medical School (10277595)36(475)pp. 366-371
Background: Colorectal cancer is the third cancer which results in death in western countries. Some of the risk factors of this cancer are age, inadequate diet, obesity, inactivity, genetic changes, etc. Considering the relationship between genetic changes with colorectal cancer is the subject of recent studies, and one of the subjects which have been focused is T241M polymorphism in Xrcc3 gene. Methods: In this case-control study, 90 patients with colorectal cancer and 83 healthy people were included. Genomic DNA was extracted from peripheral blood, and genotype distribution in the region of polymorphism was studied. By designing of primers, the considered area was amplified and genotyped using polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) method. Findings: We observed the association between T241M polymorphism and colorectal cancer. Besides, there was a meaningful relationship between aging and family history with this cancer. On the other hand, smoking was not associated with colon cancer, but it also showed a significant role in the incidence and metastasis of rectal cancer. Conclusion: The relationship between T241M polymorphism and increased risk of colorectal cancer, along with family history of cancer, old age, and smoking were reported as risk factors for colorectal cancer. Therefore, the results of this research can show the need for further investigation in subsequent studies. © 2018, Isfahan University of Medical Sciences(IUMS). All rights reserved.
Gene (18790038)674pp. 98-103
Beta-thalassemia (β-thalassemia) is a globally genetic diseases, and is most prevalent in the Middle East, particularly in Iran. Carrier detection and prenatal diagnosis are the best ways to managing it, and to prevent new community cases from emerging. We report on a simple method for rapid detection of the worst β-thalassemia point mutation in Iran (IVS-II-1 G>A), using a nano-based ligation assay, this was performed using probes with labeled magnetic nanoparticles and quantum dots. After optimizing the technique, 50 DNA samples were genotyped with this method. We found a frequency of 72% for IVSII-1 (G˃A) mutation (42% heterozygote, and 30% mutant homozygote) with a highly sensitive nano-based ligation genotyping system, offering excellent sensitivity and specificity for point mutation detection; it has been demonstrated to be inaccurate, sensitive, cost-effective, and rapid technique for single nucleotide polymorphism (SNP) genotyping. © 2018
RSC Advances (20462069)7(41)pp. 25665-25672
A nanodiagnostic genotyping method was presented for point mutation detection directly in human genomic DNA based on ligase reaction coupled with quantum dots and magnetic nanoparticle-based probes. For this purpose, allele-specific probes, including a biotin-labeled common probe and two biotin-labeled allele-specific probes were designed for mutant and wild alleles of human beta globin gene (IVS-II-I G → A point mutation). When genomic DNA carried the mutation site, the common probe and allele-specific probe were ligated to form exponential amplified biotin-labeled fluorescence ligation products. These ligated products were captured by streptavidin-coated magnetic nanoparticles at one end and then attached to a QD 605-streptavidin conjugate at the other end to be detected fluorescently. Thereafter, the genotypes were identified conveniently according to the fluorescence color of quantum dots using a rotor-gene 6000Q real-time rotary analyzer. The results demonstrated the sensitivity and specificity percentages of this nanomolecular mutation detection method were 85.45% and 95.77% respectively. In addition, this method could be a high throughput and high sensitivity detection system that represents suitable non-PCR based nanodiagnostics for detection of other point mutations. © 2017 The Royal Society of Chemistry.
International Journal Of Fertility And Sterility (2008076X)10(4)pp. 390-394
The genetic association between cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations and male infertility due to congenital bilateral absence of vas deferens (CBAVD) is well established. Mutant CFTR, however may also be involved in the etiology of male infertility in non-CBAVD cases. The present study was conducted to estimate the frequency of ΔI507 and ΔF508 CFTR gene mutations in Iranian infertile males. We undertook the first study of association between these CFTR mutations and non-obstructive azoospermia in Iran. In this case-control study, 100 fertile healthy fathers and 100 non-obstructive azoospermia’s men were recruited from Isfahan Infertility Center (IIC) and Sari Saint Mary’s Infertility Center, between 2008 and 2009. Screening of F508del and I507del mutations was carried out by the multiplex-ARMS-PCR. Significance of differences in mutation frequencies between the patient and control groups was assessed by Fisher’s exact test. The ΔF508 was detected in three patients. No significant association was however not found between the presence of this mutated allele and infertility [OR=9.2 (allele-based) and 7.2 (individual-based), P=0.179]. None of the samples carried the ΔI507 mutation. Altogether, we show that neither delta I507 nor delta F508 is involved in this population of Iranian infertile males with non-obstructive azoospermia. © 2016 Royan Institute (ACECR). All rights reserved.
Mehraban, M.H.,
Odooli, S.,
Yousefi, R.,
Roghanian, R.,
Bashi naeini, M.M.,
Moosavi-movahedi, A.,
Ghasemi, Y. Journal of Biomolecular Structure and Dynamics (07391102)35(9)pp. 1968-1978
A vast research has been conducted to find suitable and safe carriers for vital and pH-sensitive drugs including antibiotics. This article reports the use of easily accessible and abundant purified beta-lactoglobulin (β-LG) protein as the potential carrier of widely used Kanamycin (Kana) and Ciprofloxacin (Cip) antibiotics. Spectroscopic techniques (Fluorescence, UV–vis, Circular Dichroism) combined with molecular docking were used to determine the binding mechanism of these drugs. Fluorescence studies showed moderate binding affinity with the calculated binding constants KCip = 60.1 (±0.2) × 103 M−1 and Kkana = 2.5 (±0.6) × 103 M−1 with the order of Cip > Kana. Results of UV–vis were consistent with fluorescence measurements and demonstrated a stronger complexation for Cip rather than Kana. The secondary structure of β-LG was preserved upon interaction with Kana; however, a reduction in β-sheet content from 39.1 to 31.9% was convoyed with an increase in α-helix from 12.8 to 20.5% due to complexation of Cip. Molecular docking studies demonstrated that preferred binding sites of these drugs are not the same and several amino acids are involved in stabilizing the interaction. Based on the achieved results, Kana and Cip can spontaneously bind to β-LG and this protein may serve as their transport vehicle. © 2016 Informa UK Limited, trading as Taylor & Francis Group.
Tumor Biology (14230380)36(6)pp. 4757-4762
HO-1 gene encodes heme oxygenase-1 enzyme that catalyzes the oxidation of heme to carbon monoxide (CO). It has also been suggested that cells could be protected by the enzyme against stress. A (GT)n dinucleotide repeat at HO-1 promoter is a polymorphic region and modulates gene transcription and associated with some of diseases. In this study, length of polymorphism GT tandem repeat has been determined and classified into two alleles short (≤28) and long (≥29). In present study, association between GT-repeat polymorphism at heme oxygenase-1 gene promoter and increased risk of gastric cancer and metastasis was investigated. Blood samples from 100 control individuals and 60 gastric cancer cases had taken. Genotypic frequencies of (GT)n repeat for samples were determined using PCR technique and polyacrylamide PAGE electrophoresis. At final, higher frequency alleles were sequenced. Our results show that S-allele is significantly higher in cases in comparison with control groups (p = 0/000, odds ratio (OR) = 4/154). It has been shown that individuals with S/S and S/L genotypes are at high risk of having gastric cancer (p = 0/000, OR = 3/789). Statistic data show association between SS genotype and risk of gastric cancer metastasis (p = 0.017, OR = 3.889). But, there is no significant association between clinicopathological characteristics of the patients and risk of gastric cancer metastasis (p > 0.05). Significant association was found between short allele (SS + SL genotypes) and risk of gastric cancer, and also strong association was found between SS genotype and risk of gastric cancer metastasis. © 2015, International Society of Oncology and BioMarkers (ISOBM).
Avicenna Journal Of Medical Biotechnology (20084625)7(4)pp. 173-178
Background: The risk of developing female infertility has been associated with gene polymorphisms that decrease the activity of enzymes involved in systemic Oxidative Stress (OS). In this study, PON1 L55M polymorphism for association with susceptibility to infertility was investigated among Iranian female population. Methods: Samples from 120 Iranian females [20 endometriosis; 30 Polycystic Ovary Syndrome (PCO); 70 controls] were analyzed and PCR-RFLP assay was used to determine the PON1 rs854560 (L55M) frequencies. The paraoxonase (PONase) and arilesterase (AREase) activities of PON1 enzyme were also assessed in order to investigate the association between serum PON1 activities, female infertility, and PON1 L55M polymorphism. Results: The women with a MM genotype (p=0.021; OR=2.55) showed more possibilities of experiencing infertility than those with a LM genotype (p=0.039; OR=1.91). According to LSD test, endometriosis subjects had significantly lower paraoxonase enzyme activity compared to control group (p=0.0024; CI=95%). No significant difference was found in women with PCOS for both PONase and AREase activity in comparison with control group (p=0.469; CI=95%). Furthermore, PON1 activities were the highest in LL genotype followed by LM and then MM genotype (MM
Iranian Journal of Reproductive Medicine (16806433)13(9)pp. 563-570
Background: Infertility is a health problem which affects about 10-20% of married couples. Male factor infertility is involved approximately 50% of infertile couples. Most of male infertility is regarding to deletions in the male-specific region of the Y chromosome. Objective: In this study, the occurrence of deletions in the AZF region and association between infertility and paternal age were investigated in Iranian men population. Materials and Methods: To assess the occurrence of Y chromosomal microdeletions and partial deletions of the AZF region, 100 infertile men and 100 controls with normal spermatogenesis were analyzed. AZFa, AZFb, AZFc and partial deletions within the AZFc region were analyzed using multiplex PCR method. Finally, the association between paternal age and male infertility was evaluated. Results: No AZFa, AZFb or AZFc deletions were found in the control group. Seven infertile men had deletions as the following: one AZFb, five AZFc, and one AZFab. Partial deletions of AZFc (gr/gr) in 9 of the 100 infertile men (9/100, 9%) and 1 partial AZFc deletions (gr/gr) in the control group (1/100, 1%) were observed. In addition, five b2/b3 deletions in five azoospermic subjects (5/100, 5%) and 2 partial AZFc deletions (b2/b3) in the control group (2/100, 2%) were identified. Moreover, the risk of male infertility was influenced by the paternal age. Conclusion: The results of this study suggested that the frequency of Y chromosome AZF microdeletions increased in subjects with severe spermatogenic failure and gr/gr deletion associated with spermatogenic failure. © 2015 Research and Clinical Center for Infertitlity. All rights reserved.
Iranian Biomedical Journal (2008823X)19(3)pp. 177-182
Background: β-thalassemia is the most common monogenic disorder in human. The (C→T) polymorphism at -158 upstream region of the γG-globin gene and pharmacological factors such as hydroxyurea have been reported to influence γ-globin gene expression and the severity of clinical symptoms of β-thalassemia. Methods: In the present study, 51 β-thalassemia intermediate patients were studied. Xmn1γG polymorphism genotype was determined using Tetra-Primer ARMS-PCR technique. Hemoglobin (Hb) and fetal hemoglobin (HbF) levels were determined by gel electrophoresis. Results: Of 51 patients, 35 (68.6%) patients were heterozygous (CT) and 16 (31.4%) patients were homozygous (CC). Of 30 patients under treatment by hydroxyurea, 20 (66.7%) patients were heterozygous (CT) and 10 (33.3%) patients were homozygous (CC). Our results demonstrated that in the heterozygous (CT) genotype, the Hb (9.58 ± 1.25 gm/dl) and HbF (89.30 ± 21.87) levels were significantly higher in comparison with homozygous (CC) genotype (7.94 ± 1.34 gm/dl and 70.32 ± 40.56, respectively). Furthermore, we observed that after drug usage, the Hb and HbF levels in patients with heterozygous (CT) genotype (0.7 ± 1.26 gm/dl and 5.95±14.8, respectively) raised more in comparison with homozygous (CC) genotype (0.26 ± 1.43 gm/dl and 0.8±1.31, respectively). Conclusion: Hb and HbF levels in the patients carrying T allele are increased significantly, and they also response to hydroxyurea treatment. © 2015, Pasteur Institute of Iran. All rights reserved.
Iranian Journal Of Public Health (22516085)44(3)pp. 380-387
Background: β -thalassemia, a monogenic autosomal recessive disorder, is prevalent in Middle East, particularly in Iran. In Iran, near to 20 mutations in the β-globin gene are introduced as common mutations with varying incidence frequencies in each city. Therefore, detection and screening for couples at high risk can help to solve the problems of this disease. In this study, optimized genotyping of two common mutations in Isfahan Province, IVSII-I (G-A) and FSC-8/9 insG, was performed using the T-ARMS method. Methods: In this case-control study, 10 healthy individuals and 30 patients affected by β-thalassemia major with a mean 24.76 + 4.5 years were selected from Omid Hospital in Isfahan Province. After designing tetra primers for two prevalent mutations IVSII-I (G-A) and FSC-8/9 insG, samples were genotyped using tetra-primers ARMS PCR tech-nique. Results: We have developed a sensitive single tube tetra-primers PCR assay to detect both IVSII-1 (G-A) and FS8-9 insG mutations. Moreover, we have distinguished homozygous and heterozygous forms of these mutations successful-ly. The frequency of IVSII-1 (G-A) mutation from 30 patients in Isfahan was 86.6% (33.3% heterozygote, and 53.3% mutant homozygote) and for FS8-9 insG mutation was 16.6% (13.3% heterozygote, and 3.3% mutant homozygote). Conclusion: Tetra-primers ARMS PCR could be a reliable, accurate and simple technique for genotyping SNP and different mutations. So far, no study was done on optimization methods for genotyping mutations in β-thalassemia by T-ARMS. Here, we successfully adjusted and enhanced this method for recognizing two common mutations (FSC-8/9 insG and IVSII-I (G-A)) of β-thalassemia in Isfahan population. © 2015 Iran J Public Health All Right reserved.
Cell Journal (Yakhteh) (22285806)16(3)pp. 309-314
Objective: People are usually susceptible to carcinogenic aromatic amines, present in cigarrette smoke and polluted environment, which can cause DNA damage. Therefore, maintenance of genomic DNA integrity is a direct result of proper function of DNA repair enzymes. Polymorphic diversity could affect the function of repair enzymes and thus augment the risk of different cancers. Xeroderma pigmentosum group D (XPD) gene encodes one of the most prominent repair enzymes and the polymorphisms of this gene are thought to be of importance in lung cancer risk. This gene encodes the helicase, which is a component of transcription factor IIH and an important part of the nucleotide excision repair system. Studies reveal that individuals with Lys751Gln polymorphism of XPD gene have a low repairing capacity to delete the damages of ultraviolet light among other XPD polymorphisms.
Rabiee, F.,
Forouzanfar, M.,
Ghazvini zadegan, F.,
Tanhaei, S.,
Ghaedi, K.,
Bashi naeini, M.M.,
Baharvand, H.,
Nasr-esfahani, M.H. Gene (18790038)551(2)pp. 127-137
Fibronectin type III domain-containing 5 protein (Fndc5) is an exercise hormone and its transcript profile in mouse showed high degree of expression in heart, skeletal muscle and brain. Our previous studies indicated a significant increase (approximately 10 fold) in mRNA level of Fndc5 when embryonic stem cells were differentiated into beating bodies. As a step closer to identify the involvement of Fndc5 in the process of cardiomyocyte differentiation, we generated a stably inducible transduced mouse embryonic stem cell (mESC) line that overexpressed Fndc5 following Doxycycline induction. Our results indicated that the overexpression of Fndc5 during spontaneous cardiac differentiation significantly increased not only at RNA levels for mesodermal markers but also at the transcriptional levels for cardiac progenitor and cardiac genes. These data suggest that Fndc5 may be involved in cardiomyocyte differentiation. Therefore, a new hope will be arisen for potential application of this myokine for regeneration of damaged cardiac tissues especially in cardiac failure. © 2014 Elsevier B.V.All rights reserved.
Journal of Isfahan Medical School (10277595)31(264)
Background: Hydroxyurea is a chemotherapeutic agent for treatment of cancer. This drug induces globin-γ synthesis, so it could be used for treatment of thalassemia. Several studies have been shown that treatment with hydroxyurea increases Hb and HbF levels in patients with intermediate betathalassemia. However, the efficiency of hydroxyurea treatment in patients with beta-thalassemia is unclear. In the present study, clinical response of these patients to the drug was investigated. Methods: In this prospective study, the samples were patients with beta-thalassemia intermedia admitted to Sayed-al-Shohada hospital, Isfahan, Iran, during the years 2011-13. Efficiency of hydroxyurea in 46 patients was studied by determining the changes of Hb and HbF levels before and after one year of treatment with the drug. Treatment was performed using 500 mg capsule with dosage of 20 mg/day/kg. Patients were monitored for side effects, too. Findings: After treatment, the means of Hb and HbF levels increased at a rate of 0.47 ± 1.12 g/dl and 6.04 ± 1.43 percent, respectively; where the first was statistically significant, but the latter was not. Use of drug improved the quality of the patient's condition and there was no side effect. Conclusion: According to our results, it is suggested that treatment with hydroxyurea could be effective in majority of patients with intermediate beta-thalassemia.
Journal of Isfahan Medical School (10277595)32(279)pp. 359-367
Background: Fibroblast growth factor receptor 2 (FGFR2) is a receptor of tyrosine kinase with a pivotal role in the cell growth and differentiation. FGFR2 gene was identified as susceptibility gene for breast cancer by Genome-wide associated study. rs1219648 polymorphisms in intronic region are associated with breast cancer. FGFR2 gene is amplified in 15-10% of breast tumors. Single-nucleotide polymorphisms (SNPs) of this region are involved in FGFR2 amplification. In this study, the association of rs1219648 in intron 2 region of FGFR2 gene and breast cancer was assessed. Methods: In the present study, 80 cases of breast cancer and 100 healthy controls were studied. After DNA extraction from blood, specific sequence was amplified by tetra primer ARMS-PCR (amplification-refractory mutation system-polymerase chain reaction) technique and genotype of C/T polymorphism was determined by agarose gel electrophoresis. Findings: Individuals with G/G and A/G genotype were at a significantly higher risk of breast cancer (OR = 5.32, P = 0.018). G allele frequency in case patients were greater than controls but this increase did not show significant relationship with breast cancer (P = 0.230). Conclusion: Single nucleotide polymorphism of G/A in intron 2 of the FGFR2 tyrosine kinase receptor gene may play a role as a risk factor for breast cancer susceptibility.
Iranian Journal Of Biotechnology (23222921)11(3)pp. 199-204
Background: Lung cancer is considered as one of the most frequent cancers worldwide, and has been the cause of more than one million mortalities each year. Exposure to tobacco smoke is the primary cause of most lung cancers, since it contains several thousand compounds, including more than 50 known carcinogens. However, a small fraction of individuals who are exposed to tobacco smoke develop lung cancer, therefore genetic factors may render some tobacco smokers more susceptible to cancer. Objectives: Genetic polymorphism in genes that encode metabolizing enzymes may be related to differentiated susceptibility of malignancy. CYP1B1 protein is a member of the more significant CYP1 subfamily enzymes, involved in environmental carcinogenmetabolic activation. The most studied polymorphism in CYP1B1 gene includes 4325 C→G, resulting in an amino acid change from leucine to valine amino acid. Materials and Methods: A case-control study (included 65 lung cancer cases and 80 healthy controls) was designed based on the RFLP-PCR method to estimate the possible association of this polymorphism with lung cancer susceptibility in the Iranian population. Results: Regarding the distribution of CYP1B1 L432V genotypes, there were no meaningful differences among controls and lung cancer patients, however among patients carrying the CC genotype, tobacco smokers had a considerable elevated risk for lung cancer compared to those who had the GG genotype. Conclusions: CYP1B1 L432V polymorphism has an important role in lung cancer risk. Therefore, further studies are recommended for investigation of other related CYP1B1 gene polymorphisms, their association with affective genes and regulatory factors in the Iranian population. © 2013, National Institute of Genetic Engineering and Biotechnology; Licensee Kowsar Ltd.
Journal Of Kerman University Of Medical Sciences (20082843)20(5)pp. 460-469
Background & Aims: The most significant cause of infertility in men is the genetic deletion in the azoospermia factor (AZF) region that is caused by the process of intra- and inter-chromosomal homologous recombination in amplicons. Homologous recombination could also result in partial deletions in AZF region. The aim of this research was to determine the association between the partial AZFc deletions and infertility. Methods: The blood samples were taken from 100 infertile men' who referred to the Infertility Center of Isfahan' Iran. 100 healthy matched people were also selected as the control group. The five markers of sY1201' sY1206' sY1161' sY1291, and sY1191 were applied in order to study partial deletions. Partial deletions were analyzed in AZF region using the Multiplex-STS-PCR technique. The chi-square test was conducted to check the difference between pretest and posttest. Differences were considered significant if P < 0.05. Results: 9% of studied persons showed gr/gr deletion (in the patient group). Only one case of gr/gr deletion was observed in the control group. Five patients showed b2/b3 deletion. One b2/b3 deletion was seen in the control group. The b2/b4 deletion was observed in 3 patients. In conclusion' partial deletions were observed in 14% of the patients. The statistical analysis of the gr/gr deletion in the study indicates a meaningful difference, but b2/b3 deletion does not represent a meaningful difference. Conclusion: Our results suggest that gr/gr deletions are associated with spermatogenic failure, and there is no association between b2/b3 deletion and infertility.
Rezaei, H.,
Bashi naeini, M.M.,
Khodadad, K.,
Elahi, A.,
Emami, H.,
Naddaffnia, H. Gastroenterology And Hepatology From Bed To Bench (20084234)6(1)pp. 18-24
Aim: In our study, we analyzed the allelic frequency of XPD Lys751Gln polymorphism of the XPD gene and the correlation between its variant alleles with colorectal cancer in patients and control groups. Background: Human cells are routinely exposed to mutagenic and carcinogenic aromatic amines via smoking, pollution areas and other sources. These chemicals can form DNA adducts in vivo and thus lead to DNA damage. Amongst the known genetic polymorphisms of the DNA-repair genes the xeroderma pigmentosum group D (XPD, also known as ERCC2) has been the most extensively studied most commonly. Patients and methods: This study has examined the relationship between the XPD Lys 751 Gln polymorphism and colorectal cancer in 88 patients and their 88 age and sex-matched controls. Genomic DNA from peripheral whole blood was extracted using Miller method to determine the genotype of subjects with RFLP-PCR analysis. Results: This study shows cancer patients have more of the heterozygous genotype (XPD Lys 751 Gln) compared to control group. However the results are not statistically significant. Furthermore, colorectal cancer was less common in individuals with recessive homozygous genotype (P< 0.0001). Conclusion: This study suggests that individuals with heterozygous polymorphism (Lys/Gln) may have an increased susceptibility to colorectal cancer compared to other polymorphisms (Lys/Lys and Gln/Gln). © 2013 RIGLD.
Iranian Journal of Reproductive Medicine (20082177)10(4)pp. 315-320
Background: About 10% of infertilities with obstructive azoospermia are congenital and caused by CF gene mutations. M469I mutation was observed for the first time in Taiwanese patients. This mutation not only causes CF, but also may be the origin of infertility too. Objective: In this study, we aimed in designing a rapid, reliable RFLP-PCR procedure for detection of M469I mutation. The correlation and association between M469I mutation with infertility was investigated in this study. Materials and Methods: one hundred ten patients (90 non obstructive and 20 obstructive) and 60 normal individuals were considered in this study. M469I mutation was detected using RFLP-PCR. This technique was completely designed for M469I genotyping, for the first time in our study. Amplification of the region surrounding the mutation in exon 10 of CFTR gene was then performed. RFLP analysis was carried out using the NdeI restriction enzyme. Results: All genomic DNA samples were genotyped successfully. M469I mutation was observed only in patients group. Therefore, genotype containing mutant allele (GT) has been detected only in the patients group. There was no significant correlation between GT and TT genotypes with infertility (p=0.437). Conclusion: The M469I mutation has only been observed in Exon 10 CFTR gene of infertile patients, not in the control group. This mutation causes congenital bilateral absence of vaz deferens and finally infertility. This indicates a strong association between the M469I mutation and male infertility. Therefore, this is a CF-causing CFTR mutation that could be considered as a cause of infertility.
Journal Of Research In Medical Sciences (17357136)17(SUPPL.2)
BACKGROUND: Lung cancer has remained the most prevalent malignancy worldwide. It is the fifth leading cause of cancer death in Iran. Nevertheless, during last few years a gradual permanent increase in its incidence has been reported. Although the crucial role of tobacco smoke in lung cancer initiation has long been established, it is tempting to hypothesize that genetic polymorphisms may contribute to lung cancer predisposition. CYP1A1 gene encodes the main enzyme responsible for metabolic activation of several tobacco carcinogens. CYP1A1 MspI (6235T→C) polymorphism is the most studied variation within the CYP1A1, impacts on the basal levels of metabolism and is believed to be associated with elevated lung cancer risk, mainly in Asian population. METHODS: We investigated the frequency of this genetic variation in Iranian lung cancer patients through a cross-sectional study. 65 lung cancer cases and 80 healthy controls were recruited. RESULTS: The present findings confirmed the low frequency of the variant CYP1A1*2A allele in the control group. A significant increased risk for lung cancer was observed among those who possessed heterozygous (*1/*2A) genotype (Odds ratio = 2.79, 95% CI: 1.01-7.65). Adenocarcinoma was more frequent in non-smoker group (p = 0.00064); however, no significant increased risk was observed for squamous cell carcinoma and small cell carcinoma with respect to smoking. CONCLUSIONS: heterozygous (*1/*2A) genotype may increase the risk of lung cancer.
Journal of Isfahan Medical School (10277595)30(205)pp. 1393-1402
Background: Gelatinase B is not only involved in metastasis, but also alters and processes growth factors, growth factor receptors, angiogenic factors, and other proteinases and thus affects early carcinogenesis. In other words, it has a fundamental role in initiation and development of cancer. Genetic polymorphism in the promoter of gelatinase B has been reported to be associated with the risk of several cancers including lung cancer. Methods: Genotyping of gelatinase B was carried out by taking blood samples from 172 patients with lung cancer and 100 controls using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique. The observed numbers of each gelatinase B genotype were compared with that expected for a population in Hardy-Weinberg equilibrium by χ2 test. The significance of the differences of the observed alleles and genotypes between groups was tested using the odds ratio (OR) analysis. Findings: The percentage of smokers in patients was more than controls (57.5% vs. 30%). Distribution of gelatinase B genotype was significantly associated with the risk of lung cancer [OR = 2.56; 95% confidence interval (CI) = 0.06-23.82]. Conclusion: Our results indicated that smokers who carry TT and CT+TT genotypes have 4 (OR = 3.45; 95% CI = 1.28-9.24) and 15 (OR = 14.66; 95% CI = 3.95-53.47) fold risks of lung cancer, respectively.
Journal of Isfahan Medical School (10277595)30(203)
Background: Type IV collagenase gene is capable of degrading type IV collagen which is the major structural component of basement membrane. It also increased the bioavailability of pro-angiogenic factors including vascular endothelial growth factor and transforming growth factor-β. A cytosine (C) thymine (T) single nucleotide polymorphism (SNP) at position 1562 of the type IV collagenase promoter has been reported to affect gene expression. The purpose of this study was to investigate the association between the C(-1562)T polymorphism and risk of lung cancer in different age groups in Iran population. Methods: Genotyping of C(-1562)T polymorphism in type IV collagenase gene was performed by taking out genomic DNA from blood samples of 120 patients with lung cancer and 100 age-matched controls by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). Chi-square test was used to calculate adjusted odds ratio (OR) and 95% confidence interval (95% CI). All analyses were performed in SPSS. Findings: Distribution of C(-1562)T genotype in type IV collagenase promoter was significantly associated with the risk of lung cancer in the age group of < 60 years (OR = 19.89; 95% CI = 3.21-120.60). Conclusion: Our results indicated that C(-1562)T polymorphism in type IV collagenase gene affects the risk of lung cancer in different age groups, i.e. aging less than 60 years was significantly related with initiation of lung cancer.
Journal Of Research In Medical Sciences (17357136)17(10)pp. 962-966
Background: Matrix metalloproteinases comprise a family of enzyme degrade components of extra cellular matrix. There are single nucleotide polymorphisms in the promoter regions of several genes with ability to influence cancer susceptibility. The aim of this study was to analyze association between MMP3 promoter polymorphisms and colorectal cancer occurrence and progression. Materials and Methods: In this case-control study 120 colorectal cancer patients and 100 controls were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) on the genomic deoxyribonucleic acid (DNA). The patients group was divided into different subgroups: a subgroup without metastatic activity (M-) and a subgroup that had developed metastasis (M+). Results: There was a significant difference in frequency of the MMP-3 genotype between cases and controls (χ2 =16.17; P=0.0003). The 5A homozygote in patients and controls was significantly different. The frequency of the 5A allele among affected patients (67.91%) was significantly higher than among the healthy controls (49%; χ2 =16.17, P=0.00005). At the time of diagnosis, individual who was carrying the 5A allele was more represented in the M+ subgroup than in M- subgroup (χ2 =7.49; P=0.006, OR=3.86; 95% CI, 1.43-10.33). The difference between M- and controls did not observe statistically significant (χ2 =0.009; P=0.92). Conclusions: Our results suggest that the presence of 5A polymorphism at the MMP-3 promoter region may favor the growth and the metastasis process in colorectal cancer patients and could be looked at as a risk factor for a worse prognosis.
Journal of Isfahan Medical School (10277595)29(137)
Background: Colorectal cancer is the third cause of cancer death in western countries. Age, inadequate diet, obesity, inactivity and genetic changes are some of the risk factors of colorectal cancer. Corelation of genetic diversity in homologous recombination repair system with cancer was evaluated in many recent studies. This study was done to investigate the correlation of T241M polymorphism in Xrcc3 gene and colorectal cancers. Methods: In this cross-sectional study after collecting blood samples and extraction genomic DNA, genotype distribution of the polymorphism was determined by RFLP-PCR (Restriction fragment lengh-polymerase-Polymerase chain reaction) method. Finding: A significant corelation between T241M polymorphism with colorectal cancer was seen. Age and family history were also corelated with this cancer. Although, there was no statistically relationship between smoking status and colon cancer, but it showed correlation with rectum cancer and it has been also observed that the most occurance of metastatic activity is in the rectum. Conclusion: According to our study T241M polymorphism in Xrcc3 gene from homologous recombination repair systm could be a suitable factor for early diagnosis of colorectal cancer especially rectum and its co-operation with smoking status.
Journal of Isfahan Medical School (10277595)29(142)
Background: Infertility is one of the important human problems. One of the main genetic factors of infertility is the deletions in the chromosome Y' which is reported in 10-15% of men with severe azoospermia and oligospermia. Three regions in azoospermia factor (AZFa), (AZFb) and (AZFc) are specified as the spermatogenetic regions. In this study we investigated the effect of changes in Yq11.223 (DAZ) region in chromosome Y on infertility. Methods: In this study the blood samples of 100 infertile men who referred to the Infertility Center of Isfahan and 100 healthy people, as the control group were taken. The genomic DNA was extracted from blood samples. The analyses were performed by applying the polymerase chain reaction (PCR) technique in AZF region of Yq11.223. Finding: In this study 70 azoospermia and 30 oligospermia patients were investigated. The deletions in AZF region of Yq11.223 were recognized in 7 azoospermia patients but there were not detected in oligospermia patients and the control group. It was also observed that the intensity of bands were changed comparing between azoospermia patients. Conclusion: Deletion frequency in the region of Yq11.223 was observed in 7 of 100 (7%) infertile Oligospermia and Azoospermia men. Our results confirm the effect of DAZ in the man's infertility.
Journal of Mazandaran University of Medical Sciences (17359260)21(84)pp. 23-31
Background and purpose: Polymerase chain reaction (PCR) is a rather quick and accurate method employed for gene detection and isolation. Primer designing is an important issue in this technique and plays a critical role in considering both the genome properties and cloning of the isolated genes. Streptomycin antibiotic is produced by Streptomyces griseus using str gene cluster with more than 25 genes. This gene cluster contains StrR gene encoding a specific protein regulator of this cluster. The pathway specific transcriptional activator then induces transcription of other genes in the str gene cluster. In this study, the researchers aimed at isolating promoterless StrR2 gene and then cloning it. Materials and methods: To isolate promoterless StrR gene, a set of primers (StrR2) was designed. One pair of these primers (St Nes) detected the StrR from the genome. Moreover, Nested-PCR was then used to detect amplified StrR2 gene. Sites for BamHI and XbaI were designed in other primers (Str nP1and Str nP2). These primers not only amplified the StrR2 gene but also created restriction enzyme sites in the amplified fragments. Results: Using PCR, the promoterless StrR2 gene was amplified and its structure was confirmed. This gene was successfully cloned, too. The correct structure of the recombinant plasmids was confirmed using different techniques such as gel electrophoresis, PCR and restriction digestion analysis. Conclusion: Using this vector, one can subclone the promoterless StrR2 gene in the Streptomyces expression vectors containing inducible promoters.
Cell Journal (Yakhteh) (22285806)13(3)pp. 179-186
Objective: The clavulanic acid regulatory gene (claR) is in the clavulanic acid biosynthetic gene cluster that encodes ClaR. This protein is a putative regulator of the late steps of clavulanic acid biosynthesis. The aim of this research is the molecular cloning of claR, isolated from the Iranian strain of Streptomyces clavuligerus (S. clavuligerus). Materials and Methods: In this experimental study, two different strains of S. clavuligerus were used (PTCC 1705 and DSM 738), of which there is no claR sequence record for strain PTCC 1705 in all three main gene banks. The specific designed primers were subjected to a few base modifications for introduction of the recognition sites of BamHI and ClaI. The claR gene was amplified by polymerase chain reaction (PCR) using DNA isolated from S. clavuligerus PTCC 1705. Nested-PCR, restriction fragment length polymorphism (PCR-RFLP), and sequencing were used for molecular analysis of the claR gene. The confirmed claR was subjected to double digestion with BamHI and ClaI. The cut claR was ligated into a pBluescript (pBs) vector and transformed into E. coli. Results: The entire sequence of the isolated claR (Iranian strain) was identified. The presence of the recombinant vector in the transformed colonies was confirmed by the colony-PCR procedure. The correct structure of the recombinant vector, isolated from the transformed E. coli, was confirmed using gel electrophoresis, PCR, and double digestion with restriction enzymes. Conclusion: The constructed recombinant cassette, named pZSclaR, can be regarded as an appropriate tool for site directed mutagenesis and sub-cloning. At this time, claR has been cloned accompanied with its precisely selected promoter so it could be used in expression vectors. Hence the ClaR is known as a putative regulatory protein. The overproduced protein could also be used for other related investigations, such as a mobility shift assay.
Journal of Biological Research (Greece) (1790045X)13pp. 113-118
Matrix metalloproteinase-9 (MMP-9) in blood is a promising new tumor marker. Previously, a correlation between C/T-1562 MMP-9 polymorphism and tumor progression of breast cancer was reported. In the present study, we examined the association between the C/T polymorphism and plasma MMP-9 level in breast cancer patients. This study included 124 breast cancer patients, 52 of which with initial breast cancer and 72 with progressive breast cancer. Different allelic genotypes of MMP-9 promoter polymorphism were determined by PCR-RFLP and the plasma MMP-9 concentrations were measured using ELISA. Plasma MMP-9 levels were significantly increased in breast cancer patients with the MMP-9 promoter T allele compared with patients with the MMP-9 promoter C allele (p < 0.001). The plasma MMP-9 levels were correlated with progression of breast cancer and lymph node involvement (p = 0.002). According to our findings, individuals with MMP-9 promoter T allele are at a risk of higher plasma MMP-9 and they are more susceptible to breast cancer.
Reproductive Sciences (19337191)17(6)pp. 585-589
Blood matrix metalloproteinase 9 (MMP-9) level is a promising new diagnostic marker. The aim of the current caseg-control study is to investigate the correlation between the serum MMP-9 level with occurrence and progression of breast cancer. The serum MMP-9 level was investigated by gelatin zymography in 100 patients with breast cancer and 120 healthy participants. The average value of MMP-9 activity was significantly higher in patients with breast cancer than in control ( P <.002), this value correlates with tumor stage ( P =.005) and tumor size ( P =.012). Our results suggest serum MMP-9 level is a potential diagnostic marker for predicting breast cancer occurrence and progression. © 2010 The Author(s).
Iranian Journal Of Biotechnology (23222921)7(2)pp. 112
The claR of Streptomyces clavuligerus in the clavulanic acid gene cluster encodes a transcriptional regulator that controls clavulanic acid biosynthesis. The main goal of this study was isolation and molecular detection of the claR gene and its cloning in the Streptomyces specific vector (pMA:: hyg). By cinsideration of the claR gene's start codon, the specific primers were designed. After genomic DNA extraction from S. clavuligerus, the claR gene was amplified by Polymerase Chain Reaction (PCR). The structure of the amplified claR was confirmed by nested-PCR, PCR-restriction fragment length polymorphism (PCR-RFLP), and sequencing. A ligation mixture was prepared with the isolated claR gene and cut pMA::hyg vector. Escherichia coli competent cells were finally transformed with the ligation mixture. Presence of the recombinant vector in the transformed colonies was then confirmed by the colony-PCR procedure. The claR gene was also isolated from S. clavuligerus DSM41826, cloned and sequenced in the same manner. The pMA::hyg vector is a shuttle vector, which exists as a multicopy plasmid in E. coli, and as an integrative plasmid in Streptomyces. Therefore, the newly constructed vectors of this study can be regarded as an appropriate tool for site-directed mutagenesis and gene replacement strategies in S. clavuligerus.
International Journal of Integrative Biology (discontinued) (09738363)6(1)pp. 33-37
Matrix metalloproteinase-9 (MMP-9) gene plays an important role in several cancers development and progression including breast cancer. A single nucleotide substitution (C→T) in the MMP-9 promoter results in high expression of this gene and so affects susceptibility of tumor invasion and metastasis in some cancers. The aim of this study was to investigate the influence of C/T polymorphism on the occurrence and progression of breast cancer. This case-control study included 180 breast cancer patients and 100 healthy age-matched controls. Polymorphism in the promoter region (C-1562T) was genotyped by polymerase chain reaction-restriction fragment length polymorphism assay and sequencing. After data analyzing, a correlation was observed between the T allele (OR=3.27; P=0.004) and breast cancer occurrence. And also a high significant association was found between the occurrence of the T allele and progression and invasion of breast cancer (OR=5/85; P=0.000). The results suggest that the T allele of the C/T MMP-9 promoter polymorphism can be associated with the tumor development, progression and invasive phenotype of breast cancer, therefore it could be considered as a progression marker in this disease. © IJIB, All rights reserved.
Korbekandi, H.,
Abedi, D.,
Pourhosein, M.,
Bashi naeini, M.M.,
Hejazi m., ,
Narimousaei m., ,
Kabiri m., Biotechnology (discontinued) (1682296X)7(1)pp. 112-117
We aimed to optimize esterase production of Candida rugosa lipase (CRL). Active culture of C. rugosa (DSM 2031) was revived and the culture medium containing the most frequently used ingredients was optimized using a fraction of factorial design method, Taguchi. Temperature and pH of the culture was also optimized using one factor at a time method. The optimum combination of the major medium ingredients, in order of their magnitude, was (g L-1): Corn Steep Liquor (CSL) powder, (40), triolein (glyceril trioleate) (10), glucose (0) and oleic acid (2). The optimum temperature and pH were 30°C and 7, appropriately. Using this combination and conditions, esterase activity of the enzyme preparation was increased up to 9 U mL-1, which was equivalent to 20611 U mL-1 of Sigma® lipase lipolytic activity. © 2008 Asian Network for Scientific Information.
Cancer Investigation (07357907)26(8)pp. 836-842
Interstitial collagenas-1 degrades a variety of extracellular matrix components. A single Guanine insertion polymorphism in the promoter has been found that influences on the transcription and expression level of the gene. It is suggested that this polymorphism may enhance susceptibility to some types of cancer. Therefore, this case-control study evaluated the association of this genotype polymorphism with susceptibility to initiation and invasion of colorectal cancer. For this reason, whole blood samples were obtained from 150 CRC patients and 100 control subjects in Tehran. Genomic DNA was extracted and genotyped by PCR-RFLP method. We showed that 2G allele and 2G/2G genotype had higher frequencies in patients (60% and 39%, respectively) than in controls (47% and 23%, respectively). The CRC patients were divided into two groups: with metastasis (M+) and without metastasis (M-) groups. The 2G allele was more frequent in M+ group compared with control group. However, no significantly difference was observed between M-group and control (χ2 = 0.48, P = 0.78 for 2G/2G genotype). Further stratification analyses showed that only gender (OR = 2.58, 95% CI = 0.89-7.52 for women and OR = 4.12, 95% CI = 1.62-10.42 for men) and smoking (OR = 3.03, 95% CI = 1.28-7.16 for non-smokers and OR = 4.09, 95% CI = 1.18-4.15 for smoker) may modify the risk of colorectal invasion related to 2G/2G genotype. Furthermore, individual with 2G/2G genotype seems to spread metastasis, 3 years earlier than those who were 1G/1G and 1G/2G. In conclusion, to our knowledge, the present epidemiological study for the first time indicates the relationship of 2G/2G genotype polymorphism with invasion risk of colorectal cancer in subgroups of gender and smoking, especially in smoker men. Copyright © Informa Healthcare USA, Inc.
Iranian Journal Of Biotechnology (23222921)6(1)pp. 45-49
In the human genome, chromosome 11 contains a cluster of matrix metalloproteinase (MMP) genes. Single nucleotide polymorphisms in the promoter region of MMP genes are important for MMP expression. A common adenine deletion polymorphism (5A) at position -1171 of the MMP-3 gene promoter (5′-AAAAAACCAT-3′ change to 5′-AAAAACCAT-3′) facilitates transcriptional factor binding and MMP-3 promoter activity. A case-control study was performed including 120 breast cancer patients (60 patients with metastatic activity and 60 patients without metastatic activity); and 60 healthy controls. Whole blood samples were obtained from patients and healthy controls. Genomic DNA was extracted from samples and the MMP-3 5A/6A genotypes were determined using PCR-RFLP. MMP-3 genotype distributions between patients and controls were similar (OR= 0.89, 95%CI, 0.43-1.84, P= 0.047). It was observed that the 5A allele was more frequent among patients with metastatic activity than controls (OR= 2.9, 95%CI, 0.94-8.9, P= 0.074). Therefore, the 5A polymorphism in the MMP-3 promoter showed correlation with the metastasis group than patients without metastasis; both at the time of diagnosis. However our results do not show evidence for correlation between 5A/6A polymorphism and breast cancer susceptibility.
Pakistan Journal of Biological Sciences (18125735)10(18)pp. 3079-3084
The genetics of streptomycin production is well characterized in Streptomyces griseus. More than 25 clustered genes encode proteins involved in biosynthesis, regulation and transport functions. StrR, the pathway specific transcriptional activator or regulator that located in this cluster, then induces transcription of other streptomycin production genes by binding multiple sites in the gene cluster. We aim to put strR gene in to different multicopy and integrated expression vector specifically designed for Streptomyces. To start with, the isolated strR gene was ligated into pBluescript (pBs) vector and transformed into different strains of Escherichia coli. The correct structure of the recombinant plasmid, isolated from transformed E. coli, was confirmed using gel electrophoresis, PCR and double digested with restriction enzymes BamHI and EcoRI. Finally the plasmid map, named pFDstrR. This unique vector has a much expanded Multiple Cloning Site (MCS), which makes it suitable for different purposes of gene cloning and also site directed mutagenesis or gene targeting. This gene will be lifted up and transfer into different varieties of Streptomyces specific vectors in order to make different transgenic or genetically manipulated Streptomyces. © 2007 Asian Network for Scientific Information.
Journal of Biological Sciences (discontinued) (18125719)7(7)pp. 1092-1101
Homologous recombination repair starts with Double-strand Breaks (DSBs) followed by crossing-over and recombination. The expected frequency of meiotic chromosomal exchange in the region of chromosome XII encoding ribosomal DNA in Saccharomyces cerevisiae is 3.5 to 5 events per cell per meiosis. However interchromosomal meiotic recombination in the rDNA gene is very rare, suggesting repression of DSB and crossing-over. On the other band, mitotic events such as intrachromosomal recombination producing 3 μm rDNA circles (which accumulate with cellular age) and unequal sister chromatid exchanges appear to be quite common. This study looked at the rDNA breakage in the strain ORD 1181, a rad50S mutant with SK1 background, which does a relatively fast and near synchronous meiosis. The fine analysis of the rDNA array was performed using restriction endonuclease enzymes that do not cleave within the rDNA array. The results suggest that there are at least two hot regions for chromosome breakage within the rDNA array. According to our previous studies we suggest that the DSB hot regions are in one homologue. However, there is possibility that other homologue is involving in DSB too. © 2007 Asian Network for Scientific Information.
Mutation Research - Genetic Toxicology and Environmental Mutagenesis (18793592)564(2)pp. 129-137
In the yeast Saccharomyces cerevisiae the nucleolar organiser region (NOR) is located on chromosome XII. It contains 100-200 copies of rDNA - a minimum of 20 rDNA genes in tandem - and is termed the RDN locus. Yeast cells may exist in either haploid or diploid form. There are two forms of life cycle: haploid and diploid cells double by mitosis, and diploid cells are reduced to the haploid state by meiosis. Diploid cells have two homologous chromosomes for each of the 16 chromosomes. They are usually of the same size. However, in this study it is shown that homologous chromosomes XII can become different in size due to unequal sister chromatid exchange during mitosis in 'old' cells. © 2004 Elsevier B.V. All rights reserved.
