Specific Polymerase Chain Reaction Primers Research Articles

Interleukin-6-174 promoter polymorphism and susceptibility to hepatitis B virus infection in Javanese individuals

Hepatitis B virus infection is a significant risk factor for cirrhosis and hepatocellular carcinoma. Interleukin-6, a cytokine with broad pleiotropic actions, plays important role in cellular and humoral immune responses to viral infection. The -174 G/C polymorphisms of interleukin-6 is a frequent polymorphism and may determine the outcome of viral infection. To find out the genetic association between the -174 G/C polymorphism and susceptibility to hepatitis B virus infection in Javanese individuals, blood samples from Javanese hepatitis B virus-infected individuals and from Javanese healthy individuals confirmed by serological and molecular assays were evaluated. Genomic DNA was extracted from peripheral blood and the sequence specific primer-polymerase chain reaction method was applied for genotyping. The frequencies of genotypes C/C, C/G and G/G in hepatitis B virus-infected samples were 5.3%, 31.6%, 63.1% and in controls were 20%, 38%, and 42%, respectively. The frequencies of G allele in hepatitis B virus-infected samples was higher than that of healthy samples (OR 2.4, 95%CI: 1.289-4.459, p = 0.0057). These findings suggest that the G allele was associated with susceptibility to hepatitis B virus infection in Javanese individuals.

SRNAPrimerDB: comprehensive primer design and search web service for small non-coding RNAs.

Small non-coding RNAs (ncRNAs), especially microRNAs (miRNAs) and piwi-interacting RNAs (piRNAs), play key roles in many biological processes. However, only a few tools can be used to develop the optimal primer or probe design for the expression profile of small ncRNAs. Here, we developed sRNAPrimerDB, the first automated primer designing and query web service for small ncRNAs. The primer online designing module of sRNAPrimerDB is composed of primer design algorithms and quality evaluation of the polymerase chain reaction (PCR) primer. Five types of primers, namely, generic or specific reverse transcription primers, specific PCR primers pairs, TaqMan probe, double-hairpin probe and hybridization probe for different small ncRNA detection methods, can be designed and searched using this service. The quality of PCR primers is further evaluated using melting temperature, primer dimer, hairpin structure and specificity. Moreover, the sequence and size of each amplicon are also provided for the subsequent experiment verification. At present, 531306 and 2941669 primer pairs exist across 223 species for miRNAs and piRNAs, respectively, according to sRNAPrimerDB. Several primers designed by sRNAPrimerDB are further successfully validated by subsequent experiments. sRNAPrimerDB is a valuable platform that can be used to detect small ncRNAs. This module can be publicly accessible at http://www.srnaprimerdb.com or http://123.57.239.141. Supplementary data are available at Bioinformatics online.

Polymorphism of FCGR3A gene in chronic beryllium disease.

Previously we showed that alveolar macrophages(AMs) from patients with chronic beryllium disease(CBD) and beryllium sensitization(BeS) demonstrated significantly greater cell surface CD16(encoded by the FCGR3A gene) than controls. We hypothesized that these differences were related to polymorphisms in the FCGR3A gene. This study was to determine the association between FCGR3A polymorphisms in CBD, BeS versus controls as well as clinical data, providing potential information about disease pathogenesis, risk and activity. 189 CBD/154 BeS /150 controls (92 Be-exposed-non-diseased and 58 healthy controls) were included in this study. Sequence specific primers polymerase chain reaction(PCR-SSP) was used to determine FCGR3A 158V/F polymorphisms. We found significantly higher frequencies of the 158V allele (OR:1.60 (CI: 1.17-2.19),p=0.004) and 158VV homozygotes (OR:2.97 (CI:1.48-5.97) p=0.007) in CBD versus controls. No differences were found in the frequencies of FCGR3A alleles or genotypes between BeS versus controls and CBD versus BeS. Average changes in exercise testing maximum workload(Wlm),maximum oxygen consumption(VO2m) and diffusion capacity of carbon monoxide(DLCO) demonstrated greater decline over time in those CBD cases with the 158VV gene, modelled between 10 and 40 years from first beryllium exposure. The FCGR3A V158F polymorphism is associated with CBD compared to BeS and controls and may impact lung function in CBD.

Genomic structure and expression of the porcine ACTC1 gene

A partial cDNA (~1200 bp) of the porcine ACTC1 gene was identified in the subtracted foetal hind limb muscle cDNA library (44 days of gestation; using m. biceps femoris cDNA as the driver). Using specific polymerase chain reaction (PCR) primers, a bacterial artificial chromosome (BAC) clone containing the genomic ACTC1 gene was identified and the gene was sequenced. Specific PCR primers designed from the BAC and cDNA sequences were used for amplification and comparative sequencing of ACTC1 of Pietrain and Meishan pigs. The gene is approximately 5.4 kb in length, is composed of 7 exons, and has a coding sequence containing 1134 bp. The gene was mapped using the INRA-Minnesota porcine radiation hybrid (IMpRH) panel to chromosome 1, with SW65 as the closest marker (41 cR; LOD = 7.73). Differences were observed in tissue-specific expression of ACTC1 that was studied by transcription profiling in 28 porcine tissues. Developmental differences in muscle and heart were analysed by real-time quantitative PCR (RT-qPCR). Two single nucleotide polymorphisms (SNPs) were found in intron 1. One adequately informative SNP (FM212567.1:g.901C>G) was genotyped by PCR-restriction fragment length polymorphism, and allele frequencies in eight pig breeds were calculated.

Association of tumor necrosis factor-alpha gene promoter polymorphism and its mRNA expression level in coronary artery disease

Backgroundumor necrosis factor-alpha (TNF-α) is a pro-inflammatory cytokine that may play an important role in inflammatory diseases like coronary artery disease (CAD). As a result of controversy on the impact of polymorphisms in the promoter region of TNF-α gene on transcription, this case control study was designed to evaluate the effects of TNF-α -308G > A (rs1800629) and TNF-α − 238G > A (rs361525) on mRNA expression level and risk of CAD. MethodsOverall, 379 patients with CAD and 461 healthy controls participated in this study. Subjects were matched in terms of age, sex and ethnic origin. Genotyping was performed using single specific primer-polymerase chain reaction (SSP-PCR) and TNF-α mRNA expression analysis was carried out by quantitative real time PCR. ResultsA/A and A/G genotypes in TNF-α (−308G > A) gene were significantly more frequent among patients than controls. The TNF-α (−238G > A) did not differ between CAD patients and controls (the TNF-α (−238G > A) genotypes and allele frequency showed no significant differences). Combination of AA TNF-α (−308) /GG TNF-α (−238) and AG TNF-α (−308) /AG TNF-α (−238) genotypes demonstrated higher frequencies in patients than in controls (P < .05). TNF-α mRNA expression level showed significantly higher in patients carrying A/A genotype at TNF-α (−308) compared to healthy individuals. ConclusionAlthough there was no significant association between TNF-α -238 G > A polymorphism with TNF-α expression level and pathogenesis of CAD in the studied population, The TNF-α (−308G > A) gene polymorphism was associated with its expression and development of CAD.

Molecular characterization of Alternaria alternata population isolated from Upper Egyptian tomato fruits

AbstractAlternaria alternata is the most common fungal pathogen of tomatoes in Upper Egypt. Morphological identification of this fungus is challenging; therefore, this study searched for new classification tools based on molecular techniques. Using a dilution plating method, 67 strains of A. alternata were isolated from 34 samples of rotten tomato fruits representing the Giza 80 and Edkawy cultivars. The collected strains were identified using the amplification products of the internal transcribed spacer (ITS) region, glyceraldehyde 3‐phosphate dehydrogenase (Gpd) and Alt a1, which is a gene involved in the production of most of the allergens produced by A. alternata. The screening revealed that A. alternata constituted more than half of the total fungi recovered from rotten tomatoes in this study. According to the phylogenetic analysis using these three loci, the collected strains clustered in accordance with the host cultivar type from which they had been isolated. Specific gene random primer polymerase chain reaction (SGRP‐PCR) techniques indicated that the A. alternata population in the tested region has a high genetic diversity. The pathogenicity test showed that most of the A. alternata isolates (67.2%) were highly pathogenic, and no correlation was found between the phylogenetic analysis and pathogenicity. In addition, the influence of the fungicide Disan 80% on the collected strains showed significant differences that were attributed to the source of isolation.

Using Barnacle-Specific Primers to Quantify Larva Attachment on Nano-Filled Coatings during the Early Stages of Marine Biofouling

This study tried to set up an approach to quantify the attached barnacle larvae in marine biofilms. Biofilms were collected from nano-filled coatings that previously characterized with different antifouling scores for 3-months. Quantification was achieved by 18S rDNA copy number measurement in quantitative real-time PCR (qPCR) using barnacle-species specific PCR (polymerase chain reaction) primers. Interestingly, the coatings with best anti-fouling scores were not the ones with less barnacle larva attachment in the early stage of marine biofouling, suggesting that some nano-filled coating be able to attract barnacle larvae’s attachment but can prevent their further growth either by directly inhibiting the growth or evacuating the larvae through modulated biofilm dynamics, and the relative amount of barnacle larvae attachment cannot be regarded as a prediction for long-term antifouling efficacy of a coating.

A simple and cost-effective method for screening of CRISPR/Cas9-induced homozygous/biallelic mutants

BackgroundThe CRISPR/Cas9 system is being used for genome editing purposes by many research groups in multiple plant species. Traditional sequencing methods to identify homozygous mutants are time-consuming, laborious and expensive.ResultsWe have developed a method to screen CRISPR/Cas9-induced mutants through Mutation Sites Based Specific Primers Polymerase Chain Reaction (MSBSP-PCR). The MSBSP-PCR method was successfully used to identify homozygous/biallelic mutants in Nicotiana tabacum and Arabidopsis thaliana, and we speculate that it can be used for the identification of CRISPR/Cas9-induced mutants in other plant species. Compared to traditional sequencing methods, MSBSP-PCR is simpler, faster and cheaper.ConclusionsThe MSBSP-PCR method is simple to implement and can save time and cost in the screening of CRISPR/Cas9-induced homozygous/biallelic mutants.

RANCANGAN PRIMER SPESIFIK GEN MACROPHAGE MANNOSE RECEPTOR (MMR) UNTUK POLYMERASE CHAIN REACTION (PCR) DAN SEKUENSING DEOXYRIBO NUCLEIC ACID (DNA)

The order (sequencing) determinationof DeoxyribonucleicAcid (DNA) bases is the gene's most basic information, using the methodof Polymerase Chain Reaction (PCR) as its stage. A key factor of successful detection by PCR is specific PCR primer design choice. Thedetection of diversity of Mycobacterium Mannose Receptor (MMR) gene, responsible for recognizing mannosylated antigen structureof Mycobacterium tuberculosis (M.tb) by DNA sequencing of exon 7 chromosome 10p12, related to susceptiblity for PulmonaryTuberculosis(TB), was first performed in China in 2012. The purpose of this study was to find specific primerfromboth design originatedfrom the research in China/primer I and my own design/primer IIby using Primer3 software. This study was based on 10 healthy subjectsand was a preliminary study of a research titled. The Relationship of Single Nucleotide Polymorphisms (SNPs) of Macrophage MannoseReceptor Gene to Pulmonary Tuberculosis Cases. The examination materials consist of 3 mL of EDTA blood and DNA extraction from itsbuffy coat. The resulting DNA was processed by PCR to amplify MMR gene with primer I and II. The primer I successfully amplified DNAfragments up to 780bp while primer II only 329 bp. The MMR gene DNA sequencing analysis was performed on the amplification resultof both kinds primers by using DNA Baser and Ensembl−BLAST software. The results were different, DNA sequencing result by using theprimer I was found in several chromosomes and also in several loci. Whereas, by using the primer II, it was only found in chromosome10 and in the same locus. Based on this study, it can be concluded that the specific primer design is one of the most important factorsin the success of DNA sequencing.

A multi-tool approach to assess microalgal diversity in lichens: isolation, Sanger sequencing, HTS and ultrastructural correlations

AbstractLichen thalli represent the most conspicuous examples of fungal-algal interactions. Studies that describe phycobiont diversity within entire thalli are based mainly on Sanger sequencing. In some lichen species, this technique could underestimate the intrathalline coexistence of multiple microalgae. In this study different multi-tool approaches were applied to two lichen taxa,Circinaria hispidaandFlavoparmelia soredians, to detect algal coexistence. Here, we combined Sanger sequencing, a specific polymerase chain reaction (PCR) primer, 454-pyrosequencing, phycobiont isolation and ultrastructural characterization. Furthermore, we compared pyrenoid ultrastructural features of lichenized phycobionts with microalgae isolated in culture. An improved methodology was used to isolate and propagate phycobionts which, in combination with fast genetic identification, resulted in a considerable reduction in time and cost to complete the process. This isolation method, coupled with a specific PCR primer, allowed for the detection of coexisting algae inC. hispida(fourTrebouxialineages). 454-pyrosequencing detected only a fraction of such diversity, while Sanger sequencing identified only the primary phycobiont. Ultrastructural features of the isolated algae were observed by transmission electron microscopy; the maintenance of the pyrenoid characteristics suggested the existence of differentTrebouxialineages. InF. sorediansa singleTrebouxialineage was identified using all these approaches.In cases of lichens with algal coexistence, a combination of different molecular and ultrastructural approaches may be required to reveal the underlying algal diversity within a single thallus. The approach proposed in this study provides information about the relationship between molecular and ultrastructural data, and represents an improvement in the delimitation of taxonomic features which is needed to recognize intrathallineTrebouxiadiversity.

Role of TNF-Alpha and IL10 genes in the development and clinical manifestations of psoriatic arthritis

ObjectiveTo evaluate the impact of polymorphisms of TNF-alpha (TNFA) and IL10 genes and their association with clinical phenotypes of psoriatic arthritis (PsA). Materials and methodsThe study included 104 unrelated Venezuelan individuals, grouped into 52 patients with PsA, who fulfilled the CASPAR criteria, and 52 healthy individuals with no family history of psoriasis. The polymorphisms of the TNFA and IL10 genes were determined by Single Specific Primer-Polymerase Chain Reaction (SSP-PCR). ResultsThe GA genotype and A allele of the TNFA-238G/A polymorphism appears to confer protection against the development of PsA (OR: 0.31, 95% CI: 0.92–1.05, p=0.02). The GA genotype of the TNFA-308G/A polymorphism is associated with a late onset age of PsA (GA=60±13.17 years vs. GG=43.55±14.29 years, p=0.002), and the GG genotype of the IL10 -1082A/G polymorphism with a longer time interval between the onset of psoriasis and the development of PsA (GG=27.4±24.11 years, GA=5.47±7.23 years, AA=7.86±8.51 years, p=0.001). The CC genotypes of IL10-819 C/T and IL10-592 C/A confer risk of damage to distal interphalangeal joints (OR: 4.79, p=0.026). ConclusionsThe TNFA-238G/A polymorphism plays an important role in the development of PsA in mixed-race Venezuelans. Likewise, TNFA-308 G/A, IL10 -1082 A/G, -819C/T, -592C/A polymorphisms may modify the clinical expression of PsA.

Association of LTC4S gene rs730012 single nucleotide polymorphism with childhood asthma

Objective To investigate the association between the single nucleotide polymorphism(SNP) of leukotriene C4 synthase(LTC4S) rs730012 in the childhood asthma. Methods Sequence specific primers-polymerase chain reaction was used to assess the genetic polymorphism of LTC4S rs730012 in 105 asthma children with different order of severity and 128 non-asthma children in our hospital in the northeast of China to analyse the association between the SNP of LTC4S rs730012 and susceptibility, clinical phenotype in asthma children. Results (1)In case group, genotype frequencies of A/C, A/A and C/C were 71.4%、25.7%、2.9%, allele frequencies for A, C were 84.3%, 15.7%.In control group, the genotype frequencies of A/A, A/C, C/C, were 70.3%, 28.9%, 0.8%, allele frequencies for A, C were 84.8%, 15.2%.No significant difference was found in AA genotype and C allele frequencies between case and control group(χ2=0.035、0.020, P both>0.05). (2)C/C genotype or C allele frequencies in moderate-severe asthma group were significantly higher than the mild asthma group(χ2=5.859、5.641, P both<0.05); (3) SaO2 of A/A group was significantly higher than A/C and C/C group(t=2.976, Pboth<0.05), and FeNO and obstructive ventilatory disorder incidence rate in A/C, C/C group were higher than A/A group, the differences were statistically significant(t=2.946、χ2=5.564, P both<0.05). Conclusion The SNP of LTC4S rs730012 is associated with the order of severity, SaO2, FeNO, pulmonary function in asthma children of northeast China.However, the rs730012 is not associated with the susceptibility for asthma. Key words: LTC4S; Gene polymorphism; Childhood asthma; Severity

Common variant of 5,10-methylenetetrahydrofolate reductase may increase risk of coronary artery disease in the Iranian population

BackgroundCoronary artery disease (CAD) is the most prevalent form of cardiovascular disease that is caused by the formation of plaque in the arteries walls. Both genetic and environmental factors play an important role in the development of CAD. AimThe aim of this study was to determine the association of 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism with CAD in an Iranian population. Subjects and methodsIn this case/control study, sequence specific primer-polymerase chain reaction (SSP-PCR) method was used for genotyping of 310 patients with CAD and 367 healthy controls. ResultsFrequency of C/T genotype was significantly higher in the patients group than the control group (P = .03, OR: 1.6, 95% CI: 1.04–2.47). Based on the assumption that T is a risk allele, dominant model compares C/C genotypes to C/T + T/T genotypes. A significant association was observed in MTHFR C677T when the effect of the polymorphism was considered under a dominant genetic model (OR = 1.59; 95% CI = 1.03–2.46; P = .02). Evaluating genotype frequencies in 4 different ethnic groups (Fars, Turkmen, Sistani, and others) demonstrated significant statistical association of C/T genotype in Fars sub-groups (OR = 1.8; 95% CI = 1.11–3.06; P = .01) but this association is not observed in other populations. Significant association of C/T (P = .01, OR: 2.21, 95% CI: 1.15–4.4) genotype was found in women, but this association was not observed in men. ConclusionThe results of this study showed that C/T genotype in MTHFR C677T position is a causative factor, especially in women, and might be associated with susceptibility to CAD in the Iranian population.

Gelatin speciation using real-time PCR and analysis of mass spectrometry-based proteomics datasets

Authentication of gelatin, an ingredient in food and drug products, is of interest with respect to some regulations and religion rules. Determination of the gelatin origin is particularly challenging because of the similarity in amino acid sequences of collagen types in different species. In this study, Polymerase Chain Reaction (PCR) technique using species-specific primers and also chemometrics analysis of mass spectral data were investigated to differentiate bovine, porcine and fish source of gelatin in commercial pure gelatin and gelatin-containing food and drug products. The specific PCR primer set was optimized using real-time PCR approach to routinely determine the gelatin source. Real-time PCR is sensitive to identify gelatin origin from traces of species-specific DNA. However, source determination is impossible when DNA is denatured or removed during production process. Alongside, chemometrics methods (PLS/DA and PCA) were used for mining LC/MS data sets to obtain patterns for discrimination of gelatin origins. LC/MS data sets involve MS spectrum of tryptic gelatin peptides generated from gelatin samples. Using PLS/DA method, a discrimination model is developed for identifying origin of the extracted gelatin. The method is practical to determine the halal authenticity of gelatin in the pure and processed products.

Papel de los genes TNFA e IL10 en el desarrollo y manifestaciones clínicas de la artritis psoriásica

ObjectiveTo evaluate the impact of polymorphisms of TNF-alpha (TNFA) and IL10 genes and their association with clinical phenotypes of psoriatic arthritis (PsA). Materials and methodsThe study included 104 unrelated Venezuelan individuals, grouped into 52 patients with PsA, who fulfilled the CASPAR criteria, and 52 healthy individuals with no family history of psoriasis. The polymorphisms of the TNFA and IL10 genes were determined by Single Specific Primer-Polymerase Chain Reaction (SSP-PCR). ResultsThe GA genotype and A allele of the TNFA-238G/A polymorphism appears to confer protection against the development of PsA (OR: 0.31, 95% CI: 0.92 -1.05, P=.02). The GA genotype of the TNFA-308G/A polymorphism is associated with a late onset age of PsA (GA=60±13.17 years vs. GG=43.55±14.29 years, P=.002), and the GG genotype of the IL10 -1082A/G polymorphism with a longer time interval between the onset of psoriasis and the development of PsA (GG=27.4±24.11 years, GA=5.47±7.23 years, AA=7.86±8.51 years, P=.001). The CC genotypes of IL10-819C/T and IL10-592C/A confers risk of damage to distal interphalangeal joints (OR: 4.79, P=.026) ConclusionsThe TNFA-238G/A polymorphism plays an important role in the development of PsA in mixed-race Venezuelans. Likewise, TNFA-308 G/A, IL10 -1082 A/G, -819C/T, -592C/A polymorphisms may modify the clinical expression of PsA.

Human platelet antigens are associated with febrile non-hemolytic transfusion reactions

BackgroundFebrile non-hemolytic transfusion reaction (FNHTR) is the most common type of transfusion reactions, and it could be reduced by transfusing patients with leukocyte-poor blood products. However, FNHTR still occur in certain patients transfused with leukocyte-poor red blood cell (LPR) products. It is examined whether human platelet antigen (HPA) could be a potential membrane antigen that plays a role in FNHTR. MethodsA total of 120 inpatient subjects who transfused with LPR (60 in FNHTR group, 60 in control group) were typed for HPA-2, HPA-3, and HPA-15 using sequence specific primer-polymerase chain reaction (SSP-PCR) and electrophoresis. ResultsHPA-2 unmatched rate between donors and patients in FNHTR group was 18%, and only 3% unmatched rate was observed in control group (p=0.0082). FNHTR group was further classified according to the imputability. There was a significant difference (p=0.0041) between FNHTR (probable imputability, infection) group and control group, and more significant difference (p=0.0008) was seen between FNHTR (probable imputability, febrile neutropenia) group and control group. ConclusionsThose results indicated that HPA-2 might play roles on inducing FNHTR in patients suffering from infectious diseases and febrile neutropenia. HPA-2 genotyping between donors and recipients might be worth integrating in pre-transfusion testing to increase transfusion safety.

Human Leukocyte Antigen Class I and II Variants in Yemeni Patients with Chronic Renal Failure.

Human leukocyte antigens (HLAs) are found to be significant genetic factors concerning the susceptibility of an individual to certain diseases. To determine the association between variants of class I (A and B) and class II (DRB1) HLA alleles and chronic renal failure (CRF), compared with healthy controls, in Yemen. A case-control study in the Urology and Nephrology Center at Al-Thawra University Hospital in Sana'a, Yemen was carried out between January 2013 and December 2015 and included 187 CRF patients, and 194 healthy controls visiting the same center for kidney donation. All CRF patients in the study were on haemodialysis. The control group was confirmed to be healthy following a clinical examination by specialist physicians. Among both patients and controls, HLA class I (A and B) and class II (DRB1) HLA typing was carried out by Sequence Specific Primers (SSP) polymerase chain reaction (PCR). There was a significant protective function for HLA-A*30 gene (CRF 9.1% vs. con 16%, p=0.045) against CRF development. There was a high frequency of HLA-A*02, HLA-B*51 and HLA-DRB1*04 alleles in both patients and controls. No HLAs were located to have a significant association with genetic tendency to CRF in the current study population, however, certain HLA alleles, for instance in HLA-A*30, could be considered protective against CRF progress.

Association assessment of Interleukine-10 gene polymorphism and its expression status with susceptibility to coronary artery disease in Iran

BackgroundThe cytokines are potent inflammatory factors that regulate each stage of atherosclerosis leading to the disease development. Interleukin-10 (IL-10) as an anti-inflammatory cytokine can develop atherosclerosis by inhibiting the synthesis of metalloproteinase. Moreover, IL-10 promotes the plaque stability by preserving the extracellular matrix and fibrous cap. AimWe evaluated the association of the two IL-10 promoter gene polymorphisms with susceptibility to coronary artery disease (CAD) in Iranian population. Subjects and methodsWe used the Sequence Specific Primer-Polymerase Chain Reaction method to determine genotypes. We also studied mRNA expression of the IL-10 gene in Iranian CAD patients using quantitative real-time PCR. ResultsThere was a significant association between IL-10(−819) T allele and IL-10(−819) T/T genotype, and CAD (p=0.041 and p=0.042 respectively). There also was a significant association between IL-10(−1082) G allele and IL-10(−1082) G/G genotype, and CAD (p=0.017 and p=0.020 respectively). Genotype T/T of IL-10(−819) polymorphism significantly associated with the two vessel disease type (p=0.017). In addition, there was a significant association between IL-10(−1082) G/G genotype and the three vessel disease type (p=0.009). IL-10 mRNA expression was significantly decreased 3.36-fold in samples with IL-10(−819) polymorphism and 1.98-fold in individuals with IL-10(−1082) polymorphism. ConclusionsOur results suggest that IL-10 gene promoter polymorphisms may influence both coronary artery disease risks and severity in Iranian patients.

Role of angiotensin converting enzyme and angiotensinogen gene polymorphisms in angiotensin converting enzyme inhibitor-mediated antiproteinuric action in type 2 diabetic nephropathy patients.

AIMTo investigate the role of genetic variants of angiotensin converting enzyme (ACE) and angiotensinogen (AGT) genes in the antiproteinuric efficacy of ACE inhibitor therapy in diabetic nephropathy (DN) patients.METHODSIn the present study, 270 type 2 diabetes mellitus patients with nephropathy were enrolled and treated with ACE inhibitor (ramipril) and followed at 6 mo for renal function and albumin excretion by estimating serum creatinine, end stage renal disease, and albumin/creatinine ratio (ACR) in urine. Genotyping of ACE I/D and AGT M235T polymorphisms were performed by using primer specific polymerase chain reaction (PCR) and PCR-RFLP techniques, respectively.RESULTSForty-eight percent of DN patients (responders) benefited with respect to proteinuria from ACE inhibitor therapy at 6 mo follow-up. A significant reduction in ACR was observed after 6 mo treatment with ACE inhibitor irrespective of whether DN patients were micro-albuminuric (≥ 30 and < 300 mg/g creatinine) or macro-albuminuric (≥ 300 mg/g creatinine) at the time of enrollment. However, macro-albuminuric patients (55%) showed better response to therapy. A reduction in urinary ACR was found independent of genotypes of ACE I/D and AGT M235T polymorphisms although macro-albuminuric patients having TT genotype showed statistically insignificant increased response (72%).CONCLUSIONACE inhibitor therapy reduced urinary ACR by ≥ 30% in 50% of DN patients and the response is independent of ACE I/D and AGT M235T polymorphisms.

Predisposition of Angiotensin-converting Enzyme Deletion/Deletion Genotype to Coronary Artery Disease with Type 2 Diabetes Mellitus in South India.

Background:Worldwide, South Asians contribute to a high proportion of coronary artery disease (CAD) burden, mainly attributed to a high prevalence of diabetes. Early identification of such high-risk individuals would enable aggressive disease modification and prevention of complications. Definition of susceptible genotypes early in the course of disease may be one such avenue for reduction in morbidity and mortality from CAD.Aim:Our study was aimed to investigate the insertion/deletion polymorphism of angiotensin-converting enzyme (ACE I/D) gene and susceptibility to CAD in patients with type 2 diabetes mellitus (T2DM) in a South Indian population.Subjects and Methods:ACE (I/D) genotyping was performed by polymerase chain reaction specific primer for 187 CAD patients and 185 age- and sex-matched controls.Results:We observed that the ACE“II” genotype was found to be significantly associated with CAD patients (odds ratio [OR] = 1.689; P = 0.028). However, multiple logistic regression analysis revealed that ACE “DD” genotype was found to be most predominant risk factor for CAD patients with T2DM (OR = 6.118; P = 0.001).Conclusion:Our results showed that ACE (I/D) genotypes and alleles presented functional dimorphism in the development of CAD and CAD with T2DM patients in South India. This finding may be extremely useful in identifying subsets of patients where early aggressive treatment of risk factors is warranted.