Polymerase Chain Reaction-restriction Length Polymorphism Research Articles

Gene Variants of Vitamin D Receptor (TaqI T > C (rs731236) and BsmI G > A (rs1544410) in Urolithiasis

Background: Urolithiasis is the most prevalent uronephrologic disorder caused by genetic, environmental factors, and metabolic defects. Objectives: The present study aimed to find a possible association between the vitamin D receptor (VDR) TaqI and the BsmI gene polymorphisms in relation to the serum levels of calcium (Ca) and vitamin D and the risk of urolithiasis. Methods: This case-control study was conducted on 68 children with urolithiasis and 67 healthy controls for the VDR gene polymorphisms using the polymerase chain reaction-restriction length polymorphism method. Results: The frequency of TaqI C allele was 36% in patients compared to 32.1% in controls (P = 0.49). A significantly higher frequency of the TaqI CC genotype was found among patients in the age group > 5 to 10 years. No significant difference was detected in the frequency of the BsmI A allele between patients (41.9%) and controls (42.5%, P = 0.91). However, a significantly lower frequency of the BsmI AA genotype was detected compared to those patients with Ca levels of > 10.8 mg/dL among patients with Ca levels of ≤ 10.8 mg/dL. Conclusions: Based on the results, a lack of an association between the VDR TaqI and BsmI polymorphisms with the risk of urolithiasis among children from Western Iran. However, a higher frequency of the TaqI CC genotype was found in the age group > 5 to 10 years. In addition, lower levels of Ca in patients were related to a significantly lower frequency of the BsmI AA genotype. The VDR gene BsmI polymorphism might affect the calcium level and the calcium metabolism in urolithiasis.

Effect of DGAT1 gene polymorphisms in coarse-haired fat-tailed lambs of different genotypes.

The aim of this study was to investigate the DGAT1 gene polymorphism and its effects on lamb weight in kazakh and tajik sheep breeds. A total of 97 blood samples were collected from purebred (еdilbay х еdilbay) and crossbred lambs (еdilbay x gissar) breеd by the Baiserke Agro Scientific and Production Center in the Talgar District of the Almaty Region of Kazakhstan. Animals were genotyped for DGAT1-AluI polymorphism using the polymerase chain reaction-restriction length polymorphism (PCR-RFLP) method. The result of PCR-RFLP showed that purebred (еdilbay х еdilbay) sheep had three genotypes (CC, CT and TT) and crossbred sheep had two genotypes (CC and CT). The predominant genotype was CC with a frequency of 0.70 and 0.58 in purebred sheep and crossbred sheep breeds, respectively. The DGAT1 gene showed no significant association with live weight of lambs at different times in both breeds studied. However, the study showed that the CC genotype produced higher live weight at day 60 in purebred sheep (CC: 33,668 kg and CT: 32,444) and at day 120 (CC: 41,487 and CT: 40,929) in crossbred lambs. The present study was the first to investigate the polymorphism and relationships between genotypes and lamb live weights for DGAT1 gene in sheep breeds, purebred and crossbred. We conclude that further comprehensive investigations should be done for the exact evidence of the effects of DGAT1/Aluı polymorphism on lamb live weights.

A significant implication of intronic SNP (rs11246213) of IRF7 gene with disease susceptibility in Kashmiri (North-Indian) patients with Systemic Sclerosis

ObjectiveA study was undertaken to investigate whether the Interferon regulatory factor (IRF7) genomic region is involved in the pathogenesis of Systemic sclerosis (SSc). MethodsWe selected two single nucleotide polymorphisms (SNPs) of IRF7, namely rs11246213 which is an intronic variant, and rs1061501 located on the promoter for genotyping. Confirmed 100 cases were tested against 100 controls for IRF7 polymorphism study by Polymerase chain reaction-restriction length polymorphism method (PCR-RFLP) method. ResultsThe mean age of SSc patients were 36.87 ± 9.4 years; 90 females and 10 males (F:M 9:1); they were 52 (52 %) limited and 48 (48 %) diffuse subtypes. The distribution of SNP rs11246213 by genotypes (GG, AG, and AA) and by allele (G, A) was significantly different between the SSc and the control group (p ≤0.0001 for genotypes; p = 0.0002 for allele). A higher frequency of AG genotypes of rs11246213 (OR = 3.5; p < 0.0001) was observed in SSc cases than those in controls. Subgroup analysis also showed that the AG genotype and the minor allele G frequency of rs11246213 both were found higher in dc SSc than those of lc SSc subtype. Furthermore, the allele G frequency was associated with interstitial lung disease (OR = 5.21; p ≤0.001) and anti-topo I positivity (OR = 2.82; p = 0.005). However, the genotypic and allelic distribution of rs1061501 among the cases and controls was found to be non-significant. ConclusionA significant association of the IRF7 (rs11246213) variant allele with clinical variables may serve as a novel biomarker for the severity of SSc and hence bear important clinical implications.

The Relationship Between Bridging Integrator 1 Gene Polymorphism and Susceptibility to Alzheimer’s Disease

Background: Alzheimer’s Disease (AD) is the most common type of dementia. The role of genetic factors in AD development remains non-demonstrated. Objectives: In this study, we aimed to investigate the association between one of the BIN1 gene’s single-nucleotide polymorphisms (SNP) rs744373 and Late-Onset Alzheimer’s Disease (LOAD) in an Iranian population in Guilan Province. Materials &amp; Methods: In this case-control study, 110 patients with LOAD and 110 unrelated healthy controls were recruited. Polymerase chain reaction-restriction length polymorphism (PCRRFLP) was performed for genotyping the BIN1 gene’s SNP rs744373. Electrophoresis was thereafter conducted using agarose gel and DNA-safe stain, and the gels were visualized under an Ultraviolet (UV) trans-illuminator. The allelic and genotypic frequencies were determined. Results: The frequency of allele T (Wild-type allele) in the control and the LOAD groups was 70.9% (n=159) and 58.6% (n=129), respectively (P=0.007). The frequency of allele C in the LOAD group (41.4%) (n=91) was significantly higher than that of the control group (29.1%) (n=64) (P=0.007). BIN’s homozygous genotype (CC) frequency was significantly higher in the LOAD group than in the control group (P=0.043). Conclusion: The rs744373 SNP of the BIN1 gene is significantly associated with the risk of developing AD in the studied population.

Association between adiponectin and ADIPOQ (rs 1501299) gene polymorphism in gestational diabetes mellitus patients

Abstract Introduction In this study, we seek to determine the association between the polymorphism of the ADIPOQ (rs1501299) gene and adiponectin levels in gestational diabetes mellitus (GDM) patients. Methods The study participants included 200 pregnant women, 100 healthy subjects as the control group, and 100 having GDM as the case group. The enzyme-linked immunosorbent assay (ELISA) was used to examine adiponectin levels. The polymerase chain reaction-restriction length polymorphism (PCR-RFLP) was also used for genotyping examination. Results Findings showed that GDM patients had significantly lower serum adiponectin levels (p=0.001). The T/T genotype ho-mozygotes prevalence for rs1501299 polymorphism was significantly lower in GDM patients. (p=0.047). Conclusion Low serum adiponectin levels are associated with GDM. The T allele of the rs1501299 (+276 G/T) polymorphism of the ADIPOQ gene protects against GDM.

Vitamin D receptor and estrogen receptor gene polymorphisms in men with type 2 diabetes: Effects on Bone Metabolism.

There is an increased fracture risk in type 2 diabetes mellitus [DM] patients independent of bone mineral density [BMD], both in men and women. Estrogen receptor [ER]-alpha and vitamin D receptor [VDR] gene polymorphisms may predispose patients to increased osteoporosis and fracture risk. This study aims to analyze the relationship of the ER-alpha gene and VDR gene polymorphisms with indicators of bone turnover and BMD in male type 2 diabetic patients. Type 2 diabetic men diagnosed with diabetes for at least one year and healthy controls were included in this cross-sectional study. BMD was measured by dual X ray absorptiometry. Gene polymorphisms were evaluated with polymerase chain reaction-restriction length polymorphism. Serum iPTH, calcium, beta-CrossLaps (cTx), osteocalcin, and free testosterone levels were also evaluated. Participants were 141 type 2 diabetic men [55 ± 8 years] and 100 healthy controls [53 ± 7 years]. BMD measurements were not statistically different between the groups. While iPTH [p < 0.05] and serum calcium levels [p = 0.03] were higher in men with type 2 DM; beta-CrossLaps [p = 0.0001], osteocalcin [p = 0.005], and free testosterone [p = 0.04] were lower than controls. The differences in terms of the frequencies of VDR Apa, Taq, Bsm, Fok and ER-alpha polymorphisms were not statistically significant between the groups. No relationship was observed between polymorphisms and BMD in both groups. VDR and ER-alpha gene polymorphisms seem to have no effect on BMD and bone turnover in men with DM.

The interaction between Alternative Healthy Eating Index and MC4R rs17782313 gene variants on central and general obesity indices in women: A cross-sectional study.

Previous studies have shown that the C allele of melanocortin 4 receptor (MC4R) rs17782313 and the Alternative Healthy Eating Index (AHEI) are separately associated with obesity. However, the present study aimed to investigate the interaction between MC4R rs17782313 variants and the AHEI and their association with central and general obesity indices, which has not been assessed previously. In total, 291 women with body mass index (BMI) ≥ 25 kg m-2 and aged 18-48 years enrolled in this cross-sectional study. All participants were assessed for body composition, anthropometric measures, dietary intake, and blood parameters. After obtaining data of dietary intake from the 147-item food frequency questionnaire, the AHEI was calculated. MC4R rs17782313 single-nucleotide polymorphisms were assessed using the polymerase chain reaction-restriction length polymorphism method. After adjustment for age, energy intake, physical activity, marital status, and economic status, the interaction between MC4R rs17782313 and the AHEI was associated with hip circumference [β = -0.41, 95% confidence interval (CI) = -0.77 to -0.05, p = 0.02], BMI (β = -0.15, 95% CI = -0.29 to -0.02, p = 0.02), fat mass (kg) (β = -0.28, 95% CI = -0.56 to -0.01, p = 0.03), visceral fat area (β = -5.68, 95% CI = -9.55 to -1.80, p = 0.004). The other measures that appear to be suggestively related to this interaction (0.05 < p < 0.07) are waist circumference, waist-to-height ratio, trunk fat (%), trunk fat (kg), fat mass (%) and fat mass index. The interaction between MC4R rs17782313 and the AHEI can be related to central and general obesity indices in overweight/obese women.

Association of Bridging Integrator 1 Gene Polymorphism with Susceptibility to Alzheimer’s Disease

Background: Alzheimer's disease (AD) is the most common type of dementia. The role of genetic factors in the development of AD has not been demonstrated yet. Objectives In this study, our aim was to investigate the association between one of the BIN1 gene’s single-nucleotide polymorphism (SNP( (rs744373) and late-onset Alzheimer's disease (LOAD) in an Iranian population in the Guilan province. Materials &amp; Methods: In this case-control study, 110 patients with LOAD, and 110 unrelated healthy controls were recruited. Polymerase chain reaction-restriction length polymorphism (PCR-RFLP) was performed for genotyping BIN1 gene’s SNP (rs744373). Electrophoresis was there after carried out using agarose gel and DNA safe stain and the gels were visualized under an ultraviolet (UV) trans-illuminator. The allelic and genotypic frequencies were determined. Results: The frequency of allele T (Wild-type allele) in the control and the LOAD groups was 70.9% (n = 159) and 58.6% (n = 129), respectively (p = 0.007). The frequency of allele C in the LOAD group (41.4%) (n = 91) was significantly higher than that of the control group (29.1%) (n=64) (p=0.007). Frequency of homozygous genotype (CC) of BIN was significantly higher in the LOAD group than in the control group (p = 0.043). Conclusion: rs744373 SNP of BIN1 gene is significantly associated with the risk of developing AD in the studied population.

Interrelation between the rs2200733 polymorphism of the ATFB5 gene and atrial fibrillation in Uzbek patients.

Atrial fibrillation (AF) is one of the most common cardiac arrhythmias and a major predictor of morbidity and mortality. AF is a polygenic and polyetiological disease. In various ethnic groups, the strongest and most independent relationship with the development of AF was found with the 4q25 locus, where the ATFB5 gene is located. An analysis of the literature data showed that the carriage of the TT genotype of the rs2200733 ATFB5 gene polymorphism is the most unfavorable genotype for the development of AF. The purpose of the study was to identify the prevalence of genotypes and alleles of the rs2200733 polymorphism of the ATFB5 gene in Uzbek patients with AF. The study included 69 Uzbek patients with paroxysmal (n=20) and persistent AF (n=49). The control group (n=30) was composed of Uzbek patients without AF. Genotyping for the carriage of allelic variants of the rs2200733 polymorphism of the ATFB5 gene was performed using the Polymerase Chain Reaction-Restriction Length Polymorphism (PCR-RFLP) method. The distribution of the C and T alleles and the CC, CT, and TT genotypes of the rs2200733 polymorphism of the ATFB5 gene in patients with AF and controls were compared. After genotyping 69 patients with AF, the following distribution of the ATFB5 gene polymorphism rs2200733 was revealed: the CC genotype was detected in 35 (50.72%) patients, the CT genotype in 25 (36.23%) patients, and the TT genotype in 9 (13.05%) patients (p<0.001, χ²=22.435). Moreover, the C allele was detected in 95 (68.8%) patients, and the T allele was detected in 43 (31.2%) patients (p<0.001, χ²=37.696). The distribution of genotypes in the control group was as follows: the CC genotype was detected in 17 individuals (56.7%), the CT genotype was detected in 12 individuals (40%), and the TT genotype was detected in 1 individual (3.3%) (p<0.001, χ²=20.100). Moreover, the C allele was detected in 46 (76.7%) patients, and the T allele was detected in 14 (23.3%) patients (p<0.001, χ²=32.033). The TT genotype of the ATFB5 gene was found to be significantly more prevalent in patients with AF than in controls (13.1% vs 3.3%, p=0.0001). The TT genotype of the rs2200733 polymorphism of the ATFB5 gene was found to be significantly more prevalent in Uzbek patients with AF than in controls.

Culture-dependent diversity profiling of spoilage yeasts species by PCR-RFLP comparative analysis.

Spoilage caused by yeasts is a constant, widespread problem in the beverage industry that can result in major economic losses. Fruit juices provide an environment that allows the proliferation of yeast. Some factories in South Africa are not equipped with laboratory facilities to identify spoilage yeasts and outsourcing becomes a prolonged process which obstructs corrective action planning. This study aimed to establish yeast diversity and apply a rapid method for preliminary identification of spoilage yeasts associated with a small-scale fruit juice bottling factory. Yeast population in the factory was determined by isolation from the production environment, process equipment and spoiled products. PCR-RFLP analysis targeting the 5.8S-ITS region and D1/D2 sequencing was used for identification. A total of 207 yeasts belonging to 10 different genera (Candida, Lodderomyces, Wickerhamomyces, Yarrowia, Zygosaccharomyces, Zygoascus, Cryptococcus, Filobasidium, Rhodotorula/Cystobasidium and Trichosporon) were isolated and identified from the production environment and processing equipment. Candida intermedia, C. parapsilosis and Lodderomyces elongisporus were widely distributed in the factory. Zygosaccharomyces bailii, Z. bisporus, Zygoascus hellenicus and Saccharomyces cerevisiae were isolated from the spoiled products. The data provided a yeast control panel that was used successfully to identify unknown yeasts in spoiled products from this factory using polymerase chain reaction-restriction length polymorphism (PCR-RFLP) comparative analysis.

Thiopurine Methyltransferase Genetic Polymorphisms and Activity and Metabolic Products of Azathioprine in Patients with Inflammatory Bowel Disease.

Thiopurine S-methyltransferase (TPMT) is a cytoplasmic enzyme that catalyzes thiopurine drugs such as 6-mercaptopurine, 6-thioguanine, and azathioprine. There is a correlation between thiopurine drug metabolism, response, and toxicity and genetic polymorphism of TPMT. The aim of this study is to assess TPMT genetic polymorphisms activity and metabolic products of AZA in patients with IBD. Blood samples were obtained from 50 IBD unrelated patients from a private laboratory. We used polymerase chain reaction-restriction length polymorphism (PCR-RFLP) and allele-specific PCRbased assays to determine the TPMT gene for the different variants. A high-performance liquid chromatography system (HPLC) was carried out to determine the whole blood 6-TGN concentration. Determination of serum TMPT activity was done by ELISA kit. In IBD patients, 46/50 (92%) subjects were homozygous for the wild-type allele (TPMT*1/*1). Mutant TPMT*1/*2 and TPMT*1/*3C alleles were found in 4/46 (8%) and 3/47 (6%) of IBD patients, respectively. TPMT*1/*3B variant was not detected in any of the IBD patients. TPMT enzyme activity was higher in wild-type than that mutant variants TPMT*1/*2 and TPMT*1/*3C, suggesting that there are statistically significant differences between 6-TG levels and polymorphisms of TMPT enzyme. 6-TG levels significantly increased in IBD patients mutant variants TPMT*1/*2 and TPMT*1/*3C. Our results showed that TPMT polymorphisms are associated with 6-TGN levels in patients using AZA. This study suggests that AZA dosage may be determined according to the high or low prevalence of a TPMT genotype. Moreover, the results present the determination of metabolite for assessing possible safe effective dosage of the drug.

The clinical effectiveness of essential oils and aromatherapy for the treatment of Acne Vulgaris (Acne): A randomised controlled trial

Hypertriglyceridemia is associated with an increased risk of cardiovascular disease. The potential favorable effects of artichoke leaf extract (ALE) on anthropometric and metabolic indices may affect by fat mass and obesity associated (FTO)-rs9939609 polymorphism. This study was designed to evaluate the effects of ALE supplementation on cardiometabolic risk factors in hypertriglyceridemic patients regarding the interaction of rs9939609-FTO polymorphism with intervention outcomes.In this double-blind placebo-controlled randomized clinical trial, 52 patients with hypertriglyceridemia randomly allocated to receive ALE (1800 mg/day as four tablets) or matching placebo (consisting of corn starch, lactose, and avicel) for 12 weeks. The measurement of anthropometric indices, fasting blood sugar (FBS), and lipid profile was performed before and after the intervention. The FTO-rs9939609 polymorphism was genotyped by polymerase chain reaction-restriction length polymorphism (PCR-RFLP). The interaction was tested using two-way ANOVA.Forty-eight patients completed the trial (intervention, n = 24, placebo = 24). ALE and placebo groups were similar in the baseline characteristics. ALE supplementation did not change anthropometric indices and metabolic parameters. However, there was a significant interaction between FTO-rs9939609 polymorphism and TC, LDL-C, and TG level response to ALE supplementation. Moreover, significant changes in TG level were observed in A allele carriers compared to subjects with TT genotype.No significant effect of ALE supplementation was shown on anthropometric and biochemical indices in Iranian hypertriglyceridemic patients. However, rs9939609 variant of FTO gene seems to affect lipid profile response to ALE supplementation. Further clinical trials with larger sample size are suggested to clarify the possible interaction between rs9939609 variant or other variants of FTO gene and ALE supplementation in hypertriglyceridemia.

Association of Activity Altering Genotypes - Tyr113His and His139Arg in Microsomal Epoxide Hydrolase Enzyme with Esophageal Squamous Cell Carcinoma

The study aimed to explore the relationship of microsomal epoxide hydrolase (mEH) exon 3 (Tyr113His) and exon 4 (His139Arg) polymorphisms and predicted mEH activity with esophageal squamous cell carcinoma (ESCC) risk. 482 histologically confirmed cases and equal number of matched controls were analyzed by polymerase chain reaction-restriction length polymorphism (PCR-RFLP). Conditional logistic regression models were used to examine the association of polymorphisms with ESCC. We noted exon 3 slow genotype (OR = 6.57; CI 3.43–12.57) as well as predicted low mEH activity (OR = 3.99; CI 2.32–6.85) was associated with the ESCC risk. Elevated ESCC risk estimates were seen in smokers independent of genotypes but the association was stronger among smokers with exon 3 variant (OR = 6.67; 3.29–13.53) and low activity (OR = 7.52; CI 3.46–16.37) genotypes. Positive family history of cancer synergistically increased ESCC risk in the individuals who harbored exon 3 (OR = 13.59; CI 5.63–32.81) or altered mEH activity genotypes (OR = 13.35; CI 5.10–34.94). Significant interaction was seen between mEH exon 3 and exon 4 genotypes (P = 0.006) and between predicted mEH activity and positive family history of cancer (P = 0.018). These findings suggest association of ESCC risk with mEH polymorphisms which get modified by tobacco smoking and positive family history of cancer.

Genomic alterations associated with HER2+ breast cancer risk and clinical outcome in response to trastuzumab.

Human epidermal growth factor receptor 2 positive (HER2+) breast cancer (BC) is an aggressive BC subtype characterized by HER2 overexpression/amplification. Genomic alterations of HER2 and others have been reported to be associated with, HER2 overexpression and prediction of trastuzumab-response. Here, we aimed at identifying germline and somatic alterations associated with HER2+ BC and evaluating their association with clinical outcome in response to trastuzumab therapy given to HER2+ BC patients. Global Sequencing Array (GSA) and polymerase chain reaction-restriction length polymorphism (PCR-RFLP) techniques were used to determine alterations in HER2 and other HER2-interacting as well as signaling-related genes in HER2+ BC. In addition, 20 formalin fixed paraffin-embedded tissue samples were also evaluated by GSA for identifying significant variations associated with HER + BC as well as response to trastuzumab therapy. A germline variant in HER2 (I655V) was found to be significantly associated with the risk of the disease (p < 0.01). A nonsense mutation in PTPN11 (K99X), a pathogenic CCND1 splice site variant (P241P), a hotspot missense mutation in PIK3CA (E542K) and a hotspot missense mutation in TP53 (R249S); were observed in 25%, 75%, 30% and 40% of the HER2+ BC tissue samples, respectively. Mutant CCND1 (P241P) and PIK3CA (E542K) were found to be significantly associated with reduced disease-free survival (DFS) in patients treated with trastuzumab (p: 0.018 and 0.005, respectively). These results indicate that HER2, PTPN11, CCND1 and PIK3CA genes are important biomarkers in HER2+ BC. Moreover, the patients harboring mutant CCND1 and PIK3CA exhibit a poorer clinical outcome as compared to those carrying wild-type CCND1 and PIK3CA.

PPARγ Pro12Ala Polymorphism Influences the Relationship between Dietary Fat Intake, Adiposity and Lipid Profile in Patients with Type 2 Diabetes.

Background: Peroxisome proliferator-activated receptor γ (PPARγ) Pro12Ala polymorphism (rs1801282) has been associated with metabolic syndrome components in some studies. Moreover, the PPARγ gene may mediate the physiological response to dietary fat intake in a ligand-dependent manner. Methods: Metabolic syndrome components (body mass index, waist circumference, and lipid profile) were determined in 290 type 2 diabetes mellitus patients in a cross-sectional study. DNA genotyping for determining PPARγ Pro12Ala polymorphism was conducted using the polymerase chain reaction-restriction length polymorphism method. A semi-quantitative food frequency questionnaire was used to assess the participants' dietary intakes in the previous year. Results: There were significant differences between the two genotype groups of PPARγ Pro12Ala polymorphism, Ala carriers (Pro/Ala+Ala/Ala) versus non-Ala carriers (Pro/Pro), in terms of mean body mass index (p=0.04) and waist circumference (p=0.02). Below the median percentage of energy from monounsaturated and polyunsaturated fatty acids, Ala carriers had a higher body mass index (p=0.01) compared to non-Ala carriers. Furthermore, a significant interaction between this single-nucleotide polymorphism and polyunsaturated fatty acids intake on serum triglyceride levels (p=0.01) was seen, and in higher polyunsaturated fatty acids intake (≥ median) Ala carriers had lower triglyceride levels than non-Ala carriers (p=0.007). Conclusions: The findings of the current study support a significant association between PPARγ Pro12Ala polymorphism and metabolic syndrome components, and they suggest that this polymorphism can modulate the biological response of dietary fat intake on body mass index and triglyceride levels.

The interaction of FTO-rs9939609 polymorphism with artichoke leaf extract effects on cardiometabolic risk factors in hypertriglyceridemia: A randomized clinical trial

ObjectiveHypertriglyceridemia is associated with an increased risk of cardiovascular disease. The potential favorable effects of artichoke leaf extract (ALE) on anthropometric and metabolic indices may affect by fat mass and obesity associated (FTO)-rs9939609 polymorphism. This study was designed to evaluate the effects of ALE supplementation on cardiometabolic risk factors in hypertriglyceridemic patients regarding the interaction of rs9939609-FTO polymorphism with intervention outcomes. MethodsIn this double-blind placebo-controlled randomized clinical trial, 52 patients with hypertriglyceridemia randomly allocated to receive ALE (1800 mg/day as four tablets) or matching placebo (consisting of corn starch, lactose, and avicel) for 12 weeks. The measurement of anthropometric indices, fasting blood sugar (FBS), and lipid profile was performed before and after the intervention. The FTO-rs9939609 polymorphism was genotyped by polymerase chain reaction-restriction length polymorphism (PCR-RFLP). The interaction was tested using two-way ANOVA. ResultsForty-eight patients completed the trial (intervention, n = 24, placebo = 24). ALE and placebo groups were similar in the baseline characteristics. ALE supplementation did not change anthropometric indices and metabolic parameters. However, there was a significant interaction between FTO-rs9939609 polymorphism and TC, LDL-C, and TG level response to ALE supplementation. Moreover, significant changes in TG level were observed in A allele carriers compared to subjects with TT genotype. ConclusionNo significant effect of ALE supplementation was shown on anthropometric and biochemical indices in Iranian hypertriglyceridemic patients. However, rs9939609 variant of FTO gene seems to affect lipid profile response to ALE supplementation. Further clinical trials with larger sample size are suggested to clarify the possible interaction between rs9939609 variant or other variants of FTO gene and ALE supplementation in hypertriglyceridemia.

The Effect of Demographic Factors and VKORC1 1639 G>A Genotypes on Estimated Warfarin Maintenance Dose in Iranian Patients Under Warfarin Therapy.

Warfarin is an anticoagulant that inhibits vitamin K-dependent clotting factors including factor (F) II, FVII, FIX and FX. Different factors can change the effect of this anticoagulant in clinic. Therefore we assessed impact of VKORC1 -1639 G>A polymorphism and demographic factors on required maintenance dose in Iranian patients under warfarin therapy. The study population included 95 patients with a mean age of 61.3 ± 12.6years. Target INR range of 2-3 was considered for these patients. The frequency of VKORC1 -1639 G>A polymorphism was assessed by polymerase chain reaction-restriction length polymorphism(PCR-RFLP). Finally the obtain data were analyzed by SPSS software. Our study revealed that 30.5%, 49.5%, and 20% of the patients had VKORC1 (G/G), (G/A), and (A/A) genotypes, respectively. Carriers of VKORC1 G/G genotype required a higher warfarin dose as compared to A/A carriers (4.48 ± 1.32 and 2.7 ± 1.16mg/day, respectively; P < 0.01). In addition, patients with higher age required lower warfarin therapeutic dose (r = - 0.3, P < 0.01). It seems that -1639 G>A polymorphism and demographic variables had significant effects on warfarin maintenance dose in Iranian patients under warfarin therapy.

PPARγ Pro12Ala and C161T polymorphisms in patients with acne vulgaris: Contribution to lipid and lipoprotein profile

PurposeThe aim of present study was to clarify the role of peroxisome proliferator-activated receptor γ (PPARγ) Pro12Ala and C161T variants in the pathogenesis of acne vulgaris (AV) and their influence on lipid and lipoprotein profile. MethodsThe present case-control study consisted of 393 individuals including 198 patients with AV (mild-, moderate-, and severe-AV) and 195 unrelated age-matched healthy individuals from Western Iran. The PPARγ Pro12Ala and C161T polymorphisms were identified using polymerase chain reaction-restriction length polymorphism method. Also, serum lipid and lipoprotein profile and fasting blood sugar (FBS) were detected in studied individuals. ResultsIn women patients with AV significantly higher serum levels of FBS, total cholesterol, low density lipoprotein-cholesterol (LDL-C) and high density lipoprotein-cholesterol compared to healthy women were detected. Neither PPARγ Pro12Ala nor C161T polymorphism was associated with the risk of AV but the Pro allele was a risk factor for AV among all men and women patients ≥20years. The variant genotype of PPARγ CG (Pro/Ala) was associated with significantly higher levels of total cholesterol and triglycerides compared to CC (Pro/Pro) genotype. We detected a significantly lower level of FBS in the presence of CT+TT genotype of PPARγ C161T compared to CC genotype. Also, carriers of PPARγ TT genotype had significantly lower serum level of total cholesterol and LDL-C compared to CC genotype. ConclusionsOur results demonstrated the association of PPARγ Pro allele with susceptibility to AV in patients ≥20years and the influence of PPARγ Pro12Ala and C161T polymorphisms on the lipid and lipoprotein profile.

Differences in the interaction between CETP Taq1B polymorphism and dietary fat intake on lipid profile of normolipedemic and dyslipidemic patients with type 2 diabetes mellitus

Dyslipidemia is one of the major complications in patients with type 2 diabetes mellitus (T2DM). Dietary fat intake and genetic factors including CETP Taq1B polymorphism could also affect lipid profile concentrations, in particular HDL-c. We decided to study the frequency of this polymorphism and its interaction with dietary fat intake on HDL-c concentration among Iranian T2DM patients with and without dyslipidemia. In this comparative study, serum samples were collected from 55 patients with dyslipidemia and 129 patients without dyslipidemia. Validated semi-quantitative FFQ was used for food consumption data. CETP Taq1B polymorphism was studied by polymerase chain reaction-restriction length polymorphism (PCR-RFLP). We used χ2 and two-way ANOVA tests for statistical analysis. The frequency of B1B1 genotype was higher in patients with dyslipidemia (p=0.01). There was no significant relationship between CETP Taq1B polymorphism and lipid profile concentrations. In patients without dyslipidemia, the interaction between the polymorphism and total fat intake on HDL-c concentration as well as TG/HDL ratio was significant (p=0.02 and p=0.009 respectively). This was more evident in B1B1 genotype. Moreover, HDL-c concentration was significantly higher in B2B2 genotype with low total fat intake. Higher total fat intake may affect the relationship between CETP Taq1B polymorphism and HDL-c concentration in patients with normolipidemic T2DM.

DHA-enriched fish oil upregulates cyclin-dependent kinase inhibitor 2A (P16INK) expression and downregulates telomerase activity without modulating effects of PPARγ Pro12Ala polymorphism in type 2 diabetic patients: A randomized, double-blind, placebo-controlled clinical trial

The present study investigated the effects of docosahexaenoic acid (DHA)-enriched fish oil supplement on telomerase activity, mRNA expression of P16INK, IL-6, and TNF-α considering Pro12Ala polymorphism in the PPARγ gene. In this double-blind randomized controlled trial, 72 PPARγ Pro12Ala polymorphism genotyped type 2 diabetic patients aged 30-70 years were randomly assigned to receive 2.4 gr of DHA-enriched fish oil or a placebo for 8 weeks. Genotyping of the Pro12Ala polymorphism in the PPARγ gene was assessed using polymerase chain reaction-restriction length polymorphism (PCR-RFLP), telomerase activity in the peripheral blood mononuclear cell (PBMC) was measured using PCR-ELISA based on the telomeric repeat amplification protocol (TRAP), and changes in the mRNA expression of P16, IL-6, and TNF-α were measured using real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). In the DHA group, telomerase activity was decreased (p=0.001) during the intervention. In addition, between-group comparisons showed significant differences in the changes in telomerase activity (p=0.003) and P16 mRNA expression (p=0.028) and non-significant differences in TNF-α and IL-6 mRNA expression. The gene*DHA interaction could not affect changes in P16, IL-6, or TNF-α mRNA expression or in telomerase activity in PBMC. Short-time DHA-enriched fish oil supplementation caused increased levels of P16 expression and a decline in telomerase activity compared with the control group without modulating the effects of Pro12Ala polymorphism on the PPARγ gene. Because of the positive correlation between P16 activity and cellular senescence, the possibility of senescence stimulation by DHA is proposed.