investigation of mutation in Exon 27 and exon 29 of MYBPC3 gene by using PCR-SSCP/HA in cardiomyopathic hypertrophy patients in chahahrmahal va bakhtiari province

Background and aims: Hypertrophy cardiomyopathy (HCM) is the most common type of heart disease with monogenic inheritance distinguished by thickening of left ventricle, contractile dysfunction and potentially fatal arrhythmias. Great progress in clarifying the genetic basis of HCM obtained. It was identified more than 900 unique mutations in 20 genes. Mutations in the gene MYBPC3 (which encodes the cardiac Myosin binding protein C) are about 40% of clinical cases. This study aimed to investigate the presence of mutations in exons 27 and 29 of MYBPC3 gene in patients with hypertrophy cardiomyopathy in Chahar Mahal and Bakhtiari province. Methods: 30 probands of hypertrophy cardiomyopathy were selected from patients referred to cardiac clinic of the Shahrekord Medical University. To extract DNA from blood samples of patients we used standard phenol – chloroform protocol. The exons 27 and 29 were amplified by Polymerase Chain Reaction and were converted to single-stranded with Single Strand Conformation Polymorphism and they electrophoresed with double-stranded samples on polyacrylamide gels. Results: Extracted DNA was electrophoresed and the attraction ratio was examined and the outcomes indicate that the DNA extraction and quality of samples are respectable. By investigation of obtained results from SSCP/HA electrophorese in Polyacrylamide, no change was found in exons 27 and 29 of MYBPC3. Conclusion: Any of the patients had no mutation in exons 27 and 29 gene MYBPC3. Based on the results of this study there is no mutation in exons 27 and 29 gene MYBPC3 of HCM patients with autosomal dominant inheritance. It concluded that exons 27 and 29 of MYBPC3 do not have any role to cause HCM in examined patients

Screening LRTOMT gene (DFNB63 locus) in patients with recessive nonsyndromic hearing loss in hormozgan province, Iran

Background: Congenital hearing loss is the most common sensory disorder is modern societies. Two modes of syndromic and nonsyndromic genetic hearing loss can be seen. Autosomal recessive nonsyndromic inheritance hearing loss (ARNSHL) is the most common form of inheritance hearing loss. So, far mapping it, 95 loci of the 41 regions of deafness genes have been identified. Despite numerous studies in this field, for DFNB63 (Leucine rich transmembrane and O-methyltransferase or LRTOMT gene), few studies have been conducted. Thus, the locus mutations can help us to better understand the genes involved in hearing loss. Methods: 90 cases of hearing loss in Hormozgan province, Iran, were studied. DNA extracted via phenol-chloroform method. Polymerase chian reaction (PCR) was performed. Then, the product was used to perform polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and polymerase chain reaction-heteroduplex analysis (PCR-HA) methods. Samples with different bands were sequenced to determine the nucleotide changes. Findings: In different exons, 8 samples (each in a specific exon) were sequenced to determine the type of changes that shift single-strand conformation polymorphism bands. No change was found in nucleotide sequencing of exons in any of the groups. Conclusion: The results showed no relationship between non-syndromic deafness and LRTOMT gene mutations. As this gene is discovered in recent years, there are few studies on it. So far, only 5 mutation of this gene have been identified in the world that can determine pale relationship of the gene and hearing loss. © 2014, Isfahan University of Medical Sciences(IUMS). All rights reserved

Study the presence of mutations in exons 30 and 33 MYBPC3 gene in patients with hypertrophic cardiomyopathy by PCR-SSCP/HA method in Chaharmahal and Bakhtiari

Background and aims: HCM is a common form of hereditary heart disease with Mendelian inheritance that is a frequent cause of sudden cardiac death in the people younger than 35 years. In more than 50% of cases the cause of HCM identified as gene mutations. Mutations in the gene MYBPC3 (which encodes the cardiac Myosin binding protein C) are about 40% of clinical cases. This study was aimed to investigate the presence of mutations in exons 30 and 33 MYBPC3 gene in patients with hypertrophic cardiomyopathy in Chaharmahal and Bakhtiari. Methods: 30 probands of hypertrophic cardiomyopathy were selected from patients referred to the cardiac clinic of the Shahrekord Medical University. DNA extraction was done by using of standard phenol - chloroform protocol from blood samples of patients. The exons 30 and 33 were amplified by PCR and were converted to single-stranded with SSCP and they electrophoresed with double-stranded sample on polyacrylamide gels. Results: All of selected patients had hypertrophic cardiomyopathy with genetic cause. Qualities of extracted DNA were certified by electrophoresis and determination of their absorption ratio. By investigation of obtained results from SSCP/HA electrophorese in polyacrylamide, we did not find any change in exons 30 and 33. Conclusion: None of the patients had mutations in exons 30 and 33 gene MYBPC3. Based on the results of this study, there is no change in exons 30 and 33 of MYBPC3 in hypertrophic cardiomyopathy patients with autosomal dominant heritage. As the result, the mentioned exons do not consider as suspicious exons for prognosis and cause of HCM in Chaharmahal and Bakhtiari province

The association of serum levels of folic acid and homocysteine in pregnant women with Pre-Eclampsia

Introduction: Pre-eclampsia is one of the main cause of death in pregnant mothers and their babies, but its cause is still unknown. Studies have shown that increased maternal homocysteine in the blood is associated with various diseases such as pre-eclampsia. For inhibition from serum homocysteine elevation, high dose folic acid can be prescribed in these cases. This study aimed to assess the association of serum levels of folic acid and homocysteine with pre-eclampsia. Methods: This case-control study was conducted on 125 women with pre-eclampsia and 125 healthy pregnant women referred to Hajar hospital of Shahrekord, Iran during 2011-2012. 5 ml of blood sample was taken and serum levels of folic acid and homocysteine were measured. Data were analyzed using SPSS software version 20 and t-test, chi-square, two-way ANOVA and correlation regresion. P value less than 0.05 was considered significant. Results: Mean levels of homocysteine were elevated in cases with pre-eclampsia compared to control group (p=0.01) (11.27±4.8 vs. 9.9±3.9 μmol/l) and mean level of folic acid were significantly lower in cases with pre-eclampsia (p=0.002) (9.96±4.3 vs. 8.39±3.6 ng/ml). BMI, age, gestational age and neonatal weight have been shown to be different statistically in two groups (p<0.001) and other variables such as education level revealed no significant differences between two groups (p=0.257). Conclusion: Mothers with high serum levels of homocysteine and folic acid and high age and BMI are more at risk of pre-eclampsia. It seems that upper dose of folic acid can prevent or decrease the risk of pre-eclampsia

Mutation screening of 3’UTR and exons 1-2 of vsx1 gene by PCR-SSCP/HA and sequencing in patients with Vernal Keratoconjuctivis (VKC) in Shahrekord

Background and Objective: Vernal Keratoconjuctivis is an immune response in relation to environmental antigens, leading to inflammation of the conjunctiva. One of the presumable genetic factors in VKC is VSX1 gene. In this study, mutations in exon 1, exon 2 and 3'UTR of VSX1 gene in patients with VKC in Shahrekord were investigated by PCR-SSCP and PCR-HA. Materials and Methods: In this cross- sectional study, peripheral blood samples of 100 patients with VKC and 100 individuals with no confirmed eye disease as control group were investigated. Genomic DNA was extracted by phenol-chloroform method and then PCR was carried out. Then, SSCP and HA were performed and the samples with shifted bands were sequenced for the type of nucleotide change. Afterwards, to investigate the observed nucleotide change, RFLP method was used. Results: Our SSCP findings revealed six patients with shifted band in exons 1 and 2 and 13 patients in 3'UTR, which were sequenced for nucleotide change. Analysis of sequencing data showed a frameshift change (g. 25057561delG) in 3'UTR. There was no change in other sequences. Conclusion: The findings of this study showed that, VSX1 gene most probably has no effective role in VKC pathogenesis in the studied population. Therefore, the role of VSX1 genes in VKC pathogens needs further investigation

Mutation screening of exons 7 and 13 of the TMC1 gene in autosomal recessive non-syndromic hearing loss (ARNSHL) in Iran

Mutation screening of exons 7 and 13 of th

Association analysis of-416G > C polymorphism of T-cell immunoglobulin and mucin domain-1 gene with asthma in Iran

TIM (T-cell immunoglobulin (Ig) and mucin domain)-1, one of the members of TIM family, expresses on Th2 cells and promotes the production of Th2 signature cytokines. This can increase a series of responses in these cells which could be one of the causes of asthma or asthma-related phenotypes. The aim of this study was to investigate whether a TIM-1 promoter single nucleotide polymorphism (SNP), -416G>C, is associated with asthma in Iranian population. In this case-control study, existence of the -416G>C polymorphism was assessed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) in 300 patients with asthma (97 atopic, 203 nonatopic) and 309 healthy volunteers. Additionally, the relationship between these polymorphism genotypes and total serum IgE levels in this Iranian population was evaluated. We discovered a significant association between the -416G>C polymorphism and atopic asthma susceptibility in the population, but this SNP showed no connection with nonatopic asthma (PC polymorphism in TIM-1 gene could be a predisposing factor for atopic asthma in Iranian population, and CC genotype of this SNP can be associated with increased level of IgE in patients with asthma in the same population

Genetic linkage analysis of DFNB93 locus in a group of families with autosomal recessive non-syndromic hearing loss in Chahar Mahal & Bakhtiari and Kohkiluyeh & Boyer Ahmad provinces of Iran

زمینه و هدف: ناشنوایی یک اختلال حسی، عصبی است و بیشترین اختلال موجود در هنگام تولد است. بیش از 60 موارد ناشنوایی ارثی است. انواع ژنتیکی آن به دو نوع سندرومی و غیر سندرومی تقسیم می شود که نا شنوایی غیر سندرومی مغلوب اتوزومی (ARNSHL) با بیشترین درصد (70) رخ می دهد. این مطالعه با هدف تعیین پیوستگی ژنتیکی به لوکوس DFNB93 در خانواده های مبتلا به ARNSHL انجام شد. روش بررسی: این مطالعه توصیفی آزمایشگاهی بر روی 40 شجره بزرگ مبتلا به ARNSHL دارای حداقل دو بیمار، والدین سالم و عمدتاً دارای ازدواج خویشاوندی و منفی از نظر جهش های ژن GJB2، از استان های چهارمحال و بختیاری و کهگیلویه و بویراحمد انجام گردید. سپس خانواده ها برای پیوستگی ژنتیکی به لوکوس DFNB93 با استفاده از نشانگرهای STR و روش PCR و سپس ژل پلی اکریل آمید بررسی شدند. یافته ها: از تعداد 40 خانواده، 1 خانواده (5/2) به لوکوس DFNB93 پیوستگی نشان داد. ارزش SLINK این خانواده 67/2 و LOD بیشینه دو نقطه ای 05/2 و LOD بیشینه چند نقطه ای 05/2 محاسبه شد. نتیجه گیری: بر اساس نتیجه پژوهش حاضر، این لوکوس احتمالاً نقش کمی در ایجاد ناشنوایی در جمعیت مورد مطالعه (دو استان) دارد ولی برای تعیین نقش دقیق تر این لوکوس در ایجاد ناشنوایی در جمعیت ایرانی، مطالعات بیشتری ضروری می باشد

The Study of SLC26A4 Gene Causing Autosomal Recessive Hearing Loss by Linkage Analysis in a Cohort of Iranian Populations.

Sensorineural non-syndromic hearing loss is the most common disorder which affects 1 in 500 newborns. Hearing loss is an extremely heterogeneous defect with more than 100 loci identified to date. According to the studies, mutations in GJB2 are estimated to be involved in 50- 80% of autosomal recessive non-syndromic hearing loss cases, but contribution of other loci in this disorder is yet ambiguous. With regard to studies, DFNB4 locus (SLC26A4) can be classified as the second cause of hearing loss. So, this study aimed to determine the contribution of this locus in hearing loss as well as the frequency of SLC26A4 gene mutations in a population in the west of Iran. In this descriptive laboratory study, we included 30 families from the west of Iran with no mutation in GJB2 gene. Linkage analysis was performed by DFNB4 (SLC26A4) molecular markers (STR). The families with hearing loss linked to this locus were further analyzed for mutation detection. SLC26A4 gene exons were amplified and analyzed using direct DNA sequencing. In studied families, 2 families displayed linkage to DFNB4 locus. Identified mutations include mutation in exon 5 (c.416 G>T) and in splicing site of exon 7 (IVS-2 A>G or c.919-2 A>G)

Human glutathione s-transferase enzyme gene variations and risk of multiple sclerosis in Iranian population cohort

Multiple sclerosis (MS) is an inflammatory disease with unknown etiology. Oxidative stress has been demonstrated to play a role in pathological and inflammatory mechanisms of MS. Cells activate antioxidant processes in response to oxidative stress. Glutathione is one of the antioxidant agents in the brain and serves as a cofactor for glutathione s-transferase (GST) enzymes for detoxifying nerve cells. Among different classes of GST, GSTM1 and GSTT1 are associated with the loss of function due to structural homozygous deletion. The aim of this study is to investigate GSTM1 and GSTT1 null genotypes in an Iranian population. In this study, 270 patients and 250 healthy controls were investigated. Patient's disabilities were assessed by Kurtzke Expanded Disability Status Scale (EDSS) and genotypes were determined by multiplex PCR. Association between genotype and MS, type of MS, gender, and inability level were surveyed. The findings demonstrated a highly significant association between the null genotypes and MS (OR = 6.89 for M1/T1). The combination of two genotypes increased the risk of MS by 6.8 times. The null genotypes were found to be more frequent in women than in men. Moreover, a significant association was observed between the null genotype and EDSS 6–10 (OR = 3.199). No significant association was noticed between MS type and the studied genotypes. According to this study, it can be proposed that people with GSTM1 and GSTT1 deletions are at a higher risk for developing MS, which can be due to a decrease in enzymatic activity and their levels in nerve cells and the brain