Association of Visfatin gene polymorphism with obesity related metabolic disorders among Pakistani population: a case control study.

In recent years, the global prevalence of obesity and its associated metabolic disorders has reached alarming levels, presenting a significant challenge to public health worldwide. Visfatin, also known as pre-B cell colony-enhancing factor (PBEF) or nicotinamide phosphoribosyltransferase (NAMPT), is an adipokine that has been implicated in various physiological processes, including glucose homeostasis, lipid metabolism, and inflammation. The main objective of this proposed study is to find out the association between visfatin genetic variants and metabolic syndrome. The sample size of the study consisted of 300 blood samples (150 control and 150 cases). This study found that the genotypic frequency of visfatin SNPs, including rs2302559 (OD: 18.222; 95% CI 10.228-32.466; p-value < 0.001) and rs1215113036 (OD: 129.40; 95% CI 44.576-375.693; p-value < 0.001) were significantly associated with metabolic syndrome. Moreover, the frequency of the mutant alleles of both visfatin SNPs was found to be higher in patients with metabolic syndrome as compared to controls. Results of the current study indicate that people with any genetic variation of Visfatin, such as rs2302559 and rs1215113036, are more likely to develop metabolic syndrome. Visfatin genetic variants are linked to an increased risk of metabolic syndrome, implying it's role in disease pathophysiology.

Interplay of "leukemia inhibitory factor receptor gene" (rs3099124) polymorphism, leukemia inhibitory factor and ovarian steroids with unexplained infertility.

OBJECTIVE OF THE STUDY: To explore the association of leukemia inhibitory factor receptor (LIFR) gene variant rs3099124, ovarian steroids, and leukemia inhibitory factor with unexplained infertility in Pakistani females. METHODOLOGY: A case-control investigation in which eighty-one (81) females with unexplained infertility and one hundred and sixty-two (162) fertile counterparts (age and body mass index compared) were recruited between October 2016 and 2018. Ten milliliters of venous blood was collected from all participants. "Genomic DNA" was taken out from lymphocytes in peripheral blood samples. "Tetra Amplification Refractory Mutation System Polymerase Chain Reaction (T-ARMS-PCR)" was constructed through software "Primer-I". Amplification was carried out by "T-ARMS-PCR" followed by subsequent sequencing for confirmation and extensive consonance. Estradiol, Progesterone and Leukemia Inhibitory Factor (LIF) were measured in serum by ELISA. RESULTS: Statistically significant difference was noticed in genotype frequency in "LIFR-gene variant; rs3099124" (χ2 = 28.222, P value < 0.01) between research participants. Although, rs "3099124" "AA" (OR = 0.000; 95%CI = 0-0) and "GA" genotypes (OR = 0.525; 95%CI = 0.226-1.22) showed non-significant safety/protection against unexplained infertility yet minor/risk allele "A" frequency was greater in women with unexplained infertility suggesting a possible explanation of implantation failure. LIF concentration varied between fertile and infertile groups (χ2 = 9.857, P < 0.05) revealing significant threat of unexplained infertility in women with decreased LIF concentration (OR = 2.316, 95%CI = 1.214-4.416). Progesterone was significantly related to unexplained infertility in both study groups (χ2 = 20.347, P < 0.05). High progesterone reduced the possibility of unexplained infertility (OR = 0.306; 95% CI = 0.166-0.567). CONCLUSION: LIFR gene variation (rs3099124) and reduced LIF secretion may cause implantation failure in women with unexplained infertility.

Associations of vitamin D receptor encoding gene variants with premenopausal breast cancer risk.

OBJECTIVE: This study evaluates whether vitamin D receptor (VDR) gene variations are associated with premenopausal breast cancer in Pakistani cohorts. METHODS: Genomic DNA was isolated from 228 breast cancer patients and 500 non-cancer controls. Six polymorphic variants (rs11568820, rs4516035, rs2228570, rs1544410, rs7975232, rs731236) of the VDR gene were genotyped using PCR-RFLP analysis. All statistical analysis was carried out on IBM-SPSS 23 at p-value <.05. Chi-square test and odds ratios (ORs) along with 95% confidence interval (CIs) were applied to evaluate the relationship between VDR gene polymorphisms and breast cancer. RESULTS: Results showed that the A/A genotype of EcoRV (OR = 2.125, 95% CI = 1.024 to 4.412) and the A/a genotype of Apa1 (OR = 6.094, 95%CI = 4.111 to 9.033) gene polymorphism had an increased risk of premenopausal breast cancer. No associations of the Bsm1 and Taq1 polymorphisms were observed in premenopausal women. Moreover, the Cdx2 GG (OR = 0.34, 95%CI = 0.192-0.602) genotype had a significant protective effect on breast cancer. However, strong LD was existed between Bsm1/Taq1 (D' = 0.757, CI = 0.67-0.82) and Apa1/Taq1 (D' = 0.695, CI = 0.6-0.77). Haplotype analysis showed no association between premenopausal breast cancer and VDR haplotypes. CONCLUSION: These analyses indicates that the VDR EcoRV A/A and Apa1 A/a genotypes may be risk factors for breast cancer development among premenopausal women.

Influence of hs-CRP, IL-6 and TNF-α and it's role in dyslipidemia and type 2 diabetes in population of Karachi, Pakistan.

The present study was conducted to estimate the prevalence of pro-inflammatory cytokine interleukin-6, highly sensitive C-reactive protein, and tumor necrotic factor alpha and evaluate the association and role of these inflammatory markers in the pathogenesis of type 2 diabetes mellitus. A retrospective case-control study was conducted in Karachi. 400 individuals participated in the study having 200 diabetic patients and 200 controls. The subjects' profile and anthropometric indices were recorded and the levels of FPG, fasting insulin, lipid profile, IL-6, and hs-CRP were determined. Insulin resistance, beta-cell function and sensitivity were calculated by HOMA analysis using the HOMA calculator. Using independent t-test BMI, percent body fats, HbA1c, FPG, and fasting insulin were found significant (p<0.05). HOMA-IR, percent beta cell, total cholesterol, triglyceride, and HDL showed significant (p<0.05) results among cases and controls. Similarly, TNF-α and hs-CRP were also found significant (p<0.05) in cases than controls. Multiple linear regression was performed to predict the values of FPG, fasting plasma insulin, and IL-6. All models were statistically significant (p<0.05). The current study reveals that inflammation is the fundamental mechanism in obesity-induced insulin resistance, and T2DM, expanded fat stores in the body, and sedentary lifestyle are involved in the alteration of metabolic processes.

Association of BDNF and SERT gene polymorphisms with depression among Pakistani population.

Brain-derived neurotrophic factor (BDNF) and serotonin transporter (SERT) is implicated in the adverse life events which lead to depression. The variation in genetic make-up of BDNF (Val66Met) and SERT (5'-HTTLPR) are potential biomarkers in the development of neuropsychiatric disorders including depression. The purpose of this study was to investigate the correlation of functional polymorphisms of BDNF and SERT genes with depression among Pakistani population. A total of 373 participants (204 cases with depressive episodes and 169 healthy controls) with age between 14 and 65yrs were recruited from Pakistani population. BDNF and SERT gene polymorphisms were genotyped using PCR-RFLP analysis. The result showed that lack of association of Val66Met (χ2: 3.596, p>0.05) and 5'-HTTLPR (χ2: 0.634, p>0.05) gene polymorphisms were found with depression. However, SERT 'SL' (OR: 1.150, 95%CI: 0.601-2.201) and BDNF 'AA' (OR: 1.651, 95%CI: 0.585-4.660) and 'GA' (OR: 2.279, 95%CI: 0.825-6.298) genotypes might be a risk genotypes for depression. Hence, it is concluded that the functional BDNF (Val66Met) and SERT (5'-HTTLPR) gene polymorphisms may not be associated with depression. Replication studies on these polymorphisms with large sample size are needed.

Association of novel stop-gained leukaemia inhibitory factor receptor gene (rs121912501) variant, leukaemia inhibitory factor and ovarian steroids with unexplained infertility among Pakistani women.

AIMS AND OBJECTIVES: Embryo implantation is a complex process that requires sequential steps at the interface of embryo interaction with decidual endometrium. Many women after experiencing multiple attempts of assisted reproductive techniques fail to get implantation because of instability of leukaemia inhibitory factor and leukaemia inhibitory factor receptor-signal transducer and activator of transcription factor 3 (LIF-LIFR STAT3) signalling cascade. Therefore, this study explores the association of ovarian steroids, LIF and LIFR stop-gained variant using the tetra primer amplification refractory mutation system-polymerase chain reaction (TARMS-PCR) with unexplained infertility (UEX-IF) among Pakistani women. MATERIALS AND METHODS: This is a case-control study, a total of 81 unexplained infertile women and 162 fertile controls (with age and BMI matched) were inducted. Serum estradiol, progesterone and LIF were determined using enzyme-linked immunosorbent assay (ELISA). T-ARMS-PCR was designed using Primer 1 software. Genomic DNA was extracted from peripheral blood and amplified using T-ARMS-PCR followed by sequencing for validation and comprehensive concordance. RESULTS: This study established differences in LIF levels (χ2  = 9.857, P < .05) between patients and controls as well as explored the decreased LIF significantly raised the risk of UEX-IF (OR = 2.316; 95% CI = 1.214, 4.416). Progesterone (P) was significantly associated with UEX-IF between fertile and infertile counterparts (χ2  = 20.347, P < .05). It was also observed that increased Progesterone reduced the risk of UEX-IF (OR = 0.306; 95% CI = 0.166, 0.567). A rapid and inexpensive method for genotyping novel LIFR gene polymorphism through T- ARMS-PCR was successfully developed. LIFR gene SNP (rs121912501) had significant association (χ2  = 200.681, P < .05) with UEX-IF. LIFR rs121912501 "TT" genotype (OR = 5.417; 95% CI = 1.868, 15.709) and "CT" genotype (OR = 3.104, 95% CI = 1.586,6.076) were at increased risk of infertility. CONCLUSION: UEX-IF can be caused by LIFR gene variation irrespective of increased P. It may open the doors for the discovery of new management plans for infertile women.

A Functional Polymorphism (rs6265, G>A) of Brain-Derived Neurotrophic Factor Gene and Breast Cancer: An Association Study.

PURPOSE: The objective of this study was to evaluate the relationship between brain-derived neurotrophic factor (BDNF) gene (Val66Met, rs6265, G>A) polymorphism and breast cancer (BC) among females of Southern Pakistan. METHODS: This case-control study consisted of 300 females (BC cases [n = 100] and controls [n = 200]) with age range of 18 to 45 years. All participants were recruited during January to December 2014 and were screened for depression using Zung depression scale. Isolation of genomic DNA (gDNA) followed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was done. All statistical analysis was carried out on IBM-SPSS version 22 at P-value <.05. Hardy-Weinberg equilibrium (HWE), Pearson chi-square, and odds ratios (ORs) with 95% confidence interval (95% CI) were calculated. RESULTS: Genotype distribution of BDNF gene polymorphism lies in the goodness-of-fit model among controls. The statistical analyses reveal a significant association between genotype frequencies (χ2 = 12.709, P-value = .002) of BDNF and BC among cases and controls. The AA genotype (OR = 5.2, 95%CI = 0.632-42.804) increases the risk of having BC. CONCLUSIONS: Our results suggest that BDNF gene polymorphism may have an association with BC risk among Pakistani females. However, the present finding needs to be replicated with greater sample size with BC risk.

Association of oral Helicobacter pylori with gastric complications.

AIM: This study was aimed to identify the presence of Helicobacter pylori (H. pylori) genes in oral mucosa and find out their relationship between oral H. pylori infection and gastric complications. METHODS: This study is a case control study consists of 567 subjects with periodontal infection (278 gastric complication cases and 289 controls normal gastric intestinal mucosa) with age range of 20-80 years. Oral health status was recorded by calculating oral hygiene index (OHI), probing depths (PD) and clinical attachment loss (CAL). Each participant provided gastric biopsy and plaque samples which were subjected to H. pylori detection. Polymerase chain reaction (PCR) with different primers specifically ß globulin, 16SrRNA, babA, cagA, ureA, ureC and vacA gene was performed which were then analyzed using gel electrophoresis. RESULTS: No significant differences (χ2 = 11.873, p value > 0.05) were observed between oral H. pylori and gastric infections/complications. However, H. pylori increase the risk of developing gastro-esophageal reflux grade II (OR = 1.458, 95%CI = 0.659-3.226), normal upper GIT mucosa with lax esophageal sphincters (OR = 1.215, 95%CI = 0.285-5.181) and duodenal ulcer/duodenitis (OR = 2.187, 95%CI = 0.225-21.278). This study also showed a significant increased risk of gastritis with babA gene. CONCLUSION: Oral pathogenic H. pylori genes may enhance the severity of the gastric infection.

Association of low penetrance vitamin D receptor Tru9I (rs757343) gene polymorphism with risk of premenopausal breast cancer.

Objective The aim of this study was to determine whether a novel polymorphism ( Tru9I) in the low penetrance vitamin D receptor (VDR) gene is associated with risk of premenopausal breast cancer (BC). Methods This case-control study included 228 patients with BC and 503 healthy women living in Pakistan who were analyzed for the VDR Tru9I (rs757343) single nucleotide polymorphism. BC cases were histopathologically confirmed, and all healthy controls were age-matched with patients (age range, 20-45 years). DNA was extracted, and the polymerase chain reaction and restriction fragment length polymorphism assays were performed. Results The VDR Tru9I polymorphism was not significantly associated with premenopausal BC. However, the risk of BC was associated with the 'uu' genotype (odds ratio [OR], 1.141; 95% confidence interval [95% CI], 0.206-6.317). Further, mutant Tru9I was significantly associated with Grade IV carcinoma (OR, 5.36; 95% CI, 1.181-24.338). Conclusion The VDR Tru9I 'uu' genotype may increase the risk of premenopausal BC.

Association between Vitamin D receptor (Cdx2, Fok1, Bsm1, Apa1, Bgl1, Taq1, and Poly (A)) gene polymorphism and breast cancer: A systematic review and meta-analysis.

The purpose of this systemic review and meta-analysis was to examine the relationship between VDR gene polymorphisms and breast cancer. Literature was searched through PubMed database, Google scholar, and the web of knowledge from December 2015 to January 2017 and consists of 34 studies (26,372 cases and 32,883 controls). All statistical measures were done using STATA version 11.2. The heterogeneity among studies was tested using I2 statistics. Mantel-Haenszel method and DerSimonian-Laird method were used to combine data from studies using both random-effect model and fixed-effect model, respectively. Potential publication bias was evaluated by Egger's test. Sensitivity analysis was also performed to evaluate the quality and consistency in results. The results of this meta-analysis revealed that VDR gene polymorphisms (Bsm1 bb vs BB; SOR = 1.18, 95% CI = 1.054-1.322, Apa1 aa vs AA; SOR = 1.18, 95% CI = 0.87-1.59, Poly (A) LL vs SS; SOR = 1.41, 95% CI = 1.06-1.88, Fok1 ff + Ff vs FF; SOR = 1.25, 95% CI = 0.896-1.759, Apa1 aa+Aa vs AA; SOR = 1.13, 95% CI = 0.95-1.35, Poly (A) LL + LS vs SS; SOR = 1.19, 95% CI = 1.00-1.43, Poly (A) L vs S; SOR = 1.18, 95% CI = 1.03-1.35) are associated with the breast cancer. Cdx2, Bgl1, and Taq1 do not show association with breast cancer. Thus, the finding of this meta-analysis concluded that VDR Bsm1, Apa1, Fok1, and Poly (A) gene polymorphisms may be susceptible for breast cancer development.