Role of MyD88-Adaptor-Like (MAL) Gene Polymorphism rs8177374 and Cytokine (IFN-γ, TNF-α, IL-10, TGF-ß) Levels in Diverse Malaria Manifestations upon P. falciparum and P. vivax Infections.

This study investigated the role of genetic variant rs8177374 in MAL/TIRAP gene in mediating the cytokine levels of IFN-γ, TNF-α, IL-10, and TGF-ß in malaria patients due to Plasmodium falciparum or P. vivax infection. The study included human blood samples collected from patients with malaria (n = 228) and healthy controls (n = 226). P. falciparum and P. vivax groups were established based on the causative species of Plasmodium. Malaria samples were divided into mild and severe malaria groups based on the symptoms that appeared in the patients, according to the WHO criteria. In a previous study, we genotyped rs8177374 via allele specific PCR strategy. In this study, cytokine levels were estimated in the blood plasma of rs8177374 genotype samples via Sandwich Enzyme Linked Immunosorbent Assay kits. Increased IFN-γ and TNF-α levels in presence of CC genotype indicates the role of CC genotype in both severe and mild malaria groups. Enhanced IL-10 levels in the CT genotype and mild malaria groups suggest a role of CT genotype and IL-10 in the mild clinical outcomes of malaria. The rs8177374 polymorphism in MAL/TIRAP plays an important role in malaria pathogenesis.

Association of rs146292819 Polymorphism in ABCA1 Gene with the Risk of Coronary Artery Disease in Pakistani Population.

Coronary artery disease (CAD) is an inflammatory heart disease characterized by the narrowing of coronary arteries. ATP-binding cassette transporter A1 (ABCA1) is a gene involved in regulation of cholesterol efflux and formation of high-density lipoprotein cholesterol (HDL-C). Present study aimed to explore the association of ABCA1 rs146292819 polymorphism with CAD development as well as its effect on serum lipid levels in the Pakistani population. Study subjects included 300 CAD patients and 300 age- and sex-matched healthy individuals. Methods involved genomic DNA extraction, amplification of rs146292819 polymorphism using allele-specific PCR, analyzing PCR product by agarose gel electrophoresis and determination of serum lipids. In this study, genotype frequencies of rs146292819 polymorphism in CAD patients were GG (43%), GT (27%), TT (30%) as compared to GG (25%), GT (31%), TT (44%) in healthy subjects. GG genotype increased the risk of developing CAD by 2.2326 times (OR 2.2326; 95% CI 1.5775-3.1597) and caused decrease in HDL-C levels by 2.6348 times. GT genotype was neither associated with CAD development (OR 0.8504; 95% CI 0.5974-1.2106) nor HDL-C levels. TT genotype lowered the risk of CAD development by 0.5381 times (OR 0.5381; 95% CI 0.3846-0.753) and protected from drop in HDL-C levels by 0.5086 times (OR 0.5086; 95% CI 0.3429-0.7544). It can be concluded that GG genotype of rs146292819 polymorphism and altered lipid profile act as risk factors in the pathogenesis of CAD in the Pakistani population.

Role of rs4986790 Polymorphism of TLR4 Gene in Susceptibility towards Malaria Infection in the Pakistani Population.

BACKGROUND: Toll-like receptors (TLRs) of the human immune mechanism play important role in the detection of invading pathogens. TLRs specifically recognize the pathogen-associated molecular patterns (PAMPs) from pathogens and start the effective response. Single nucleotide polymorphisms (SNPs) in the TLRs can mediate their functions. Present study evaluated the importance of rs4986790 polymorphism of TLR4 gene in susceptibility towards malaria, clinical outcomes of the disease and responsible species of malaria. METHODS: Blood samples of 228 malaria patients and 226 healthy volunteers were selected for the study. Sample collection was completed during Sep 2013 to Sep 2015 from different hospitals of Punjab, Pakistan. Patient's samples were divided into P. vivax group and P. falciparum group on the basis of causative species of Plasmodium. Malaria samples were also divided into mild and severe malaria group based on clinical outcomes of the disease according to WHO criteria. Healthy individuals were placed in the control group. Whole blood was used for the isolation of DNA. Genomic DNA was isolated and amplification of targeted SNP was performed using allele-specific PCR. RESULTS: Results indicate the protective role of AA genotype against the susceptibility of P. vivax infection, OR: 0.5, 95%CI: 0.285-0.876, P=0.038. CONCLUSION: rs4986790 polymorphism of TLR4 gene modulates the susceptibility towards P. vivax infection. AA genotype is found to be protective against the development of P. vivax infection in the local population of Pakistan.

Role of MyD88-adaptor-like gene polymorphism rs8177374 in modulation of malaria severity in the Pakistani population

Abstract Introduction: The present study was designed to investigate the association between rs8177374 polymorphism and malaria symptoms due to exposure of Plasmodium vivax and Plasmodium falciparum. Materials and methods: A total of 454 samples were included in the study (228 malaria patients and 226 healthy individuals). Malaria patients, divided into P. vivax and P. falciparum groups on the basis of the causative species of Plasmodium, were categorized into mild and severe on the basis of clinical outcomes according to WHO criteria. Healthy individuals were used as controls. Allele specific PCR based strategy was used for the identification of rs8177374 SNP. Results: MyD88-adaptor-like gene polymorphism was associated with susceptibility to malaria (p < 0.001). C allele frequency (0.74) was higher in the population compared to T allele frequency (0.26). CT genotype increased the susceptibility of malaria (OR: 2.661; 95% CI: 1.722-4.113) and was positively associated with mild malaria (OR: 5.609; 95% CI: 3.479-9.044, p = 0.00). On the other hand, CC genotype was associated with severe malaria (OR: 3.116; 95% CI: 1.560-6.224, p = 0.00). P. vivax infection rate was higher in CT genotype carriers compared to other genotypes (OR: 3.616; 95% CI: 2.219-5.894, p < 0.001). Conclusion: MyD88-adaptor-like/TIR domain containing adaptor protein polymorphism for single nucleotide polymorphism rs8177374 is related with the susceptibility of malaria.

Comparative Evaluation of Heavy Metals in Patients with Rheumatoid Arthritis and Healthy Control in Pakistani Population.

BACKGROUND: Exposure to heavy metals in development of many diseases has been investigated previously, specially created by oxidative stress. The etiology of Rheumatoid arthritis (RA) is still not fully understood but oxidative stress created by heavy metals may have role in development of RA. The aim of present study was to compare serum level of heavy metals in RA and healthy control individuals. METHODS: Blood samples of 100 RA patients were collected from different hospitals in district Sargodha, Punjab, Pakistan and 100 control individuals from Dec 2013 to May 2014.The serum samples were analyzed for determination of Pb, Cd, Cr and Ni through Atomic absorption spectrophotometer (AA 6600 Shimadzu). RESULTS: Statistically highly significant difference was observed between RA patients and healthy control individuals for Pb, Cd, Cr, and Ni level (P<0.01). The difference between the means of both sexes was not significant for Pb and Cd concentrations (P>0.01). For Cr the difference between the means of both sexes was statistically not significant in RA +ve patients and highly significant difference was observed between both sexes in healthy control group (P<0.01). The difference between the means of both sexes for Ni was statistically non-significant in healthy control group while significant difference was observed between both sexes in RA +ve group (P<0.05). Statistically non-significant difference for Pb, Cd, Cr and Ni level was found among the all three age groups of RA and healthy control individuals (P>0.01). CONCLUSION: Concentration of heavy metals in serum samples of RA patients and healthy control individuals differ significantly, which shows that heavy metals may contributes towards development of RA.

Role of MyD88-adaptor-like gene polymorphism rs8177374 in modulation of malaria severity in the Pakistani population.

INTRODUCTION: The present study was designed to investigate the association between rs8177374 polymorphism and malaria symptoms due to exposure of Plasmodium vivax and Plasmodium falciparum. MATERIALS AND METHODS: A total of 454 samples were included in the study (228 malaria patients and 226 healthy individuals). Malaria patients, divided into P. vivax and P. falciparum groups on the basis of the causative species of Plasmodium, were categorized into mild and severe on the basis of clinical outcomes according to WHO criteria. Healthy individuals were used as controls. Allele specific PCR based strategy was used for the identification of rs8177374 SNP. RESULTS: MyD88-adaptor-like gene polymorphism was associated with susceptibility to malaria (p<0.001). C allele frequency (0.74) was higher in the population compared to T allele frequency (0.26). CT genotype increased the susceptibility of malaria (OR: 2.661; 95% CI: 1.722-4.113) and was positively associated with mild malaria (OR: 5.609; 95% CI: 3.479-9.044, p=0.00). On the other hand, CC genotype was associated with severe malaria (OR: 3.116; 95% CI: 1.560-6.224, p=0.00). P. vivax infection rate was higher in CT genotype carriers compared to other genotypes (OR: 3.616; 95% CI: 2.219-5.894, p<0.001). CONCLUSION: MyD88-adaptor-like/TIR domain containing adaptor protein polymorphism for single nucleotide polymorphism rs8177374 is related with the susceptibility of malaria.

Association of rs1800668 polymorphism in glutathione peroxidase- 1 gene and risk of rheumatoid arthritis in Pakistani population.

OBJECTIVE: To investigate the role of glutathione peroxidase 1 (GPX1) C/T polymorphism (rs1800668) in modulating the chances of Rheumatoid arthritis (RA) in Pakistani population. METHODS: A total of 400 individuals including 200 controls and 200 patients of RA, were genotyped. Detection of rs1800668 polymorphism was carried out using PCR based amplification strategy (allele specific). RESULTS: The results for Hardy Weinberg Equilibrium (HWE) indicated that the allele frequencies for GPX1 polymorphism were not deviant from HWE in whole population under observation. The statistical analysis indicated that significant association existed between rs1800668 polymorphism and RA (p<0.01). CT genotype increased the risk of RA development by 1.8582 times (OR: 1.8582; 95% CI 1.2154 to 2.8409). CC genotype was found to have protective effect against the disease development (OR: 0.5133; 95% CI 0.3403 to 0.7742) while TT genotype was found to have association with RA development but the risk level was marginal (OR: 1.5319; 95% CI 0.6124 to 3.8322). CONCLUSION: The present finding suggests the importance of GPX1 C/T polymorphism (rs1800668) in development of RA in Pakistani population. The protective role of CC genotype against the development of RA in local population was also observed.

Role of S180L polymorphism in etiology of malaria caused by Plasmodium falciparum in a small group of Pakistani population.

The aim of our study was to investigate the role of S180L polymorphism in modulation of acquisition of malaria caused by Plasmodium falciparum in a small group of Pakistani population. A total of 133 individuals including 60 controls and 73 patients of malaria, caused by Plasmodium falciparum, were genotyped using allele-specific PCR. Ninety-two samples successfully demonstrated the PCR amplification results, while forty-one samples could not be genotyped due to failure in PCR amplification. The allele frequency for S180L polymorphism was deviant from Hardy-Weinberg equilibrium (HWE) of the population under observation. Association was found between the observed polymorphism and the occurrence of malaria caused by Plasmodium falciparum (p = 0.01). Chances of malaria caused by Plasmodium falciparum were low in CC genotype carriers in comparison to other genotypes (Odds ratio: 0.3016; 95% CI: 0.124-0.729). The present findings suggest that S180L polymorphism is important in modulating the probability of acquisition of malaria caused by Plasmodium falciparum in Pakistani population. The CC genotype plays a protective role in local population against this type of malaria.

Association of the rs10757274 SNP with coronary artery disease in a small group of a Pakistani population.

OBJECTIVE: The present study aimed to investigate the association between the rs10757274 SNP (present on locus 9p21 in the gene for CDKN2B-AS1) and coronary artery disease (CAD) in a local population of Pakistan. METHODS: It was a case-control study. An allele-specific PCR-based strategy was used for the identification of genotypes. A total of 350 samples were used for the investigation, out of which 220 samples were CAD patients and 130 samples were normal healthy individuals. Effects of parameters, like family history of CAD, smoking, presence of diabetes, and hypertension, in changing the chances of CAD were studied. Odds ratio was estimated with 95% confidence interval. RESULTS: A strong association was observed between CAD and factors, like smoking (OR: 1.666; 95% CI: 1.042-2.664), presence of hypertension (OR: 26.55; 95% CI: 15.95-44.20), diabetes (OR: 3.009; 95% CI: 1.841-4.920), and family history of CAD (OR: 4.9; 95% CI: 2.965-8.099). Results for the association between the genotype on the basis of rs10757274 showed a strong association between the GG genotype and the occurrence of CAD (OR: 9.603; 95% CI: 5.746-16.05). CONCLUSION: The present results suggest the importance of the 9p21 locus in modulating the chances of CAD.

Genetic etiology of coronary artery disease considering NOS 3 gene variant rs1799983.

Reduced production of nitric oxide due to rs1799983 single nucleotide polymorphism in nitric oxide synthase 3 gene (NOS3) may enhance the risk of coronary artery disease. The association of rs1799983 polymorphism with coronary artery disease was investigated in the local population of Pakistan. Study consisted of 376 individuals, out of which 198 were coronary artery disease patients and 178 were normal healthy individuals. Allele-specific polymerase chain reaction (PCR) based strategy was used for the detection of different genotypes of rs1799983 polymorphism. PCR amplification results were obtained for 354 samples. Frequency of T allele was higher as compared to G allele in our population. Strong association between rs1799983 and coronary artery disease was observed (p < 0.01). TT genotype was found to enhance 5.717 times the risk of coronary artery disease (odds ratio (OR): 5.717; 95% confidence interval (95% CI) 3.586-9.115). On the basis of present results, it can be concluded that rs1799983 is strongly associated with coronary artery disease in our population and TT genotype of this polymorphism enhanced the risk of coronary artery disease in Pakistani population.

Electrolyte Disturbance and the Type of Malarial Infection.

BACKGROUND: Electrolytes play an important role in the normal functioning of human body. Electrolyte imbalance and mineral disturbances is the common clinical manifestation in several infectious diseases including malaria. Malaria is a mosquito borne serious infectious disease of the world. Plasmodium vivax and P. falciparum are the main agents responsible for malaria in Pakistan. Electrolyte imbalance in malarial infection may lead towards the severity of disease. METHODS: The present study analyzed the electrolytes levels (Na, K, Ca and Mg) in malarial patients and healthy individuals. Patients were categorized into two groups, P. falciparum and P. vivax, based on causative species of Plasmodium. Study consisted of 173 individuals, out of which 73 were malarial patients and 100 were normal healthy individuals. RESULTS: Concentrations of Na, K, and Ca were low in the blood of malarial patients as compared to healthy individuals (P<0.05). No significant difference for these electrolytes exists between P. falciparum and P. vivax infected groups (P>0.05). The concentration of Mg was changed based on exposure to the type of parasite. In P. falciparum infection, the level of Mg was lower than healthy individuals was (P<0.05). Discordantly, in case of P. vivaxinfection, Mg level was higher than healthy individuals were (P<0.05). No variation was noticed in electrolytes levels due to gender differences (P>0.05). CONCLUSION: Variation in Mg levels occurs due to exposure of Plasmodium depending on its type. The levels of Na, K and Ca are also changed due to Plasmodium, regardless of its type.