Sabir Hussain

A novel deletion mutation in CENPJ gene in a Pakistani family with autosomal recessive primary microcephaly

AbstractAutosomal recessive primary microcephaly (MCPH) is a rare human genetic disorder in which the head circumference is reduced because of abnormality in fetal brain growth. To date, six loci and four genes have been identified for this condition. Our study of primary MCPH led to the identification of 33 Pakistani families with different ethnic backgrounds. Most of these families showed linkage to MCPH5 locus on chromosome 1q31. Only one family with Pashtoon origin from a remote region in Pakistan linked to MCPH6 locus on chromosome 13q12.12-q12.13. Sequence analysis of exon 11 of CENPJ gene, located at MCPH6 locus, revealed a novel four base pair deletion mutation, which is predicted to be protein truncating.

Association of Interleukin-6 Gene Promoter Polymorphism with Coronary Artery Disease in Pakistani Families

Interleukin-6 (IL-6) is a well-known inflammatory cytokine and suggested to be involved in the development of coronary artery disease (CAD). IL-6 gene expression has been investigated with controversy in CAD patients. This study investigates the association of the IL-6 gene expression with CAD, the molecular basis for the regulation of interleukin-6 expression in a Pakistani population. Our data show that the serum IL-6 levels were increased in patients with CAD compared with healthy controls and that the IL-6 gene polymorphism at -174 was more prevalent in CAD cases. There was a statistically significant association between the IL-6 gene polymorphism and CAD, which may be associated with an increased risk for the disease. Moreover, circulating IL-6 and hs-CRP levels were significantly higher in patients with CC genotype (P < 0.0001 and P < 0.0001, resp.). In a binary logistic-regression model, an independent association was found between CAD and increased serum IL-6 and hs-CRP levels and -174G>C polymorphism. This is the first report on the IL-6 expression and the IL-6 gene polymorphism in patients with CAD from Pakistan, and hence it highlights a novel risk factor for the disease.

Resistin gene promoter region polymorphism and the risk of hypertrophic cardiomyopathy in patients

Resistin, a novel cytokine, is associated with an inflammatory process and is suggested to induce hypertrophy in rat cardiomyocytes. Resistin gene expression has not been investigated in patients with hypertrophic cardiomyopathy (HCM). This study investigates resistin levels in HCM patients and healthy controls and the molecular basis for the regulation of the resistin gene (RETN) in a Pakistani population. Patients with HCM (n = 105) and healthy individuals (n = 110) were enrolled in this investigation. Serum resistin levels were determined by enzyme-linked immunosorbent assay (ELISA). RETN genotyping was performed by polymerase chain reaction (PCR) and DNA sequencing. Our data showed a statistically significant increase in resistin levels from HCM patients compared with healthy subjects (6.3 +/- 2.7 ng/mL in patients vs 3.4 +/- 2.1 ng/mL in controls, P < 0.0001). The RETN -420 C > G polymorphism was significantly high in patients with HCM compared with the control group (P < 0.001). There was a significant difference between the C and G alleles from HCM cases and controls (odds ratio [OR] = 3.54, 95% confidence interval [CI] = 2.36-5.30, P < 0.0001). Logistic-regression analysis showed that the increased resistin levels, and the RETN-420 C > G polymorphism were significantly associated with HCM. Our data suggest that the elevated resistin levels and the RETN -420 C > G polymorphism may be associated with cardiac hypertrophy in the study population.

Heritability of genetic variants of resistin gene in patients with coronary artery disease: a family-based study.

OBJECTIVE The resistin gene (RETN) -420C>G and +299G>A polymorphism was investigated in a case-control study from forty complex Pakistani families with coronary artery disease (CAD) history. Heritability of the susceptible/variant alleles was investigated from parent-offspring trios in these families. METHOD Resistin levels were determined from 239 individuals by enzyme-linked immunosorbent assay. Genotyping was performed using polymerase chain reaction and restriction fragment length polymorphism. RESULTS Elevated resistin levels were observed from CAD cases vs. controls (p<0.0001). The RETN -420C>G and +299G>A polymorphism was more prevalent in cases vs. controls (p<0.0001). The transmission disequilibrium test revealed a significant association of the -420 and +299 polymorphism with CAD (χ(2)=34.4, p<0.0001 and χ(2)=31.6, p<0.0001, respectively). CONCLUSION Elevated resistin, and the RETN -420C>G and +299G>A polymorphism may contribute to familial CAD. The -420 and +299 variant alleles are transmitted more frequently from parent to affected offspring. This is the first report on the association of the RETN +299G>A polymorphism with CAD.

TNF-alpha-308G>A polymorphism and the risk of familial CAD in a Pakistani population.

A case-control and trio-families study was performed to establish a potential association between TNF-alpha gene promoter SNPs at -308 and -238, and occurrence of CAD in a Pakistani population. In the first phase, 150 patients and 150 controls were enrolled in the case-control association study. In the second phase, heritability of susceptible alleles was investigated from 88 trio-families with CAD affected offspring. Biochemical analysis of lipids and hs-CRP was carried out spectrophotometrically, while serum TNF-alpha concentrations were determined by enzyme-linked immunosorbent assay. Genotyping of the TNF-alpha SNPs were determined by PCR-RFLP method. Elevated serum TNF-alpha and hs-CRP were observed from CAD vs. controls (P<0.0001; for both). The evaluation of TNF-alpha-308G>A polymorphism in case-control study revealed that the said SNP was significantly associated with the increased risk of CAD. The findings demonstrated a significant link between the TNF-alpha variant allele A at -308 and CAD (P=0.0035), whereas the -238 SNP was not associated with the disease. Haplotype A-G of the TNF-alpha gene at -308G>A and -238G>A showed higher frequency in the patient group compared with controls (P<0.05). Moreover, data showed preferential transmission of the disease susceptible allele A at TNF-alpha-308 from parent to affected offspring in a trio-family study (P<0.0001). The current research leads to conclusion that the TNF-alpha-308G>A polymorphism is associated with CAD in the study population. Furthermore, for the first time, we showed that the TNF-alpha-308A allele was significantly associated with the familial CAD in our high risk population.

Association between tumour necrosis-α gene polymorphisms and acne vulgaris in a Pakistani population

The cytokine tumour necrosis factor (TNF)‐α is a well‐studied potent candidate mediator that is systemically involved in a variety of inflammatory diseases. Several single nucleotide polymorphisms (SNPs) of the TNF‐α gene have been studied with regard the pathogenesis of acne vulgaris, but the results have been inconclusive.

Variants of resistin gene and the risk of idiopathic dilated cardiomyopathy in Pakistan

Background In cardiovascular disease phenotypes, a genetic factor is an important determinant of both familial and non-familial dilated cardiomyopathies. Resistin is a novel adipocyte derived peptide, associated with inflammation and suggested to be involved in contractile abnormalities of cardiomyocytes. Methods In this study, we examined the association of the RETN SNPs in − 420 and + 299 in patients with idiopathic dilated cardiomyopathy (IDCM). Patients with IDCM (n = 250) and healthy controls (n = 250) were enrolled in this study. RETN genotyping was performed by using PCR-RFLP method. Results RETN − 420C > G and + 299G > A polymorphisms were significantly more prevalent in patient group vs. controls (P < 0.0001 and P = 0.0007, respectively). GG genotype at − 420 and AA genotype at + 299 were higher in the patient group compared with healthy controls (OR = 11.4, P < 0.0001, and OR = 2.3, P = 0.030, respectively). We found that the − 420G allele increased the risk of developing IDCM in patients (P < 0.0001). Moreover, there was a significant difference between G and A alleles at RETN + 299 from IDCM cases and controls (P = 0.0032). The RETN − 420G and + 299A haplotypes were more prevalent in the patient vs. control group (P < 0.0001). Conclusion The results suggest that the RETN − 420C > G and + 299G > A polymorphisms may have a role in the pathogenesis of IDCM.

Heritability of IL-1A Gene Promoter Polymorphism in Patients With Coronary Artery Disease: A Trio-Family Study.

OBJECTIVE To investigate the potential role of IL-1A gene variations in pathogenesis of coronary artery disease (CAD) with familial history. MATERIALS AND METHODS We investigated the IL-1A-889C>T polymorphism in 335 patients with CAD and 335 healthy individuals for case-control association analysis. In this study, we also investigated the heritability of the susceptible allele from 130 trio families with CAD-affected offspring. We performed genotyping using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS The IL-1A-889 C/T polymorphism was significantly associated with CAD in patients compared with healthy controls. The minor allele T at -889 was more prevalent in cases vs controls. The results of a transmission disequilibrium test revealed a significant association between the IL-1A-889 polymorphism and CAD. The mutant genotype CT+TT was significantly associated with high levels of high-sensitivity C-reactive protein (CRP) and other relative markers from patients with CAD. CONCLUSION For the first time in the literature, to our knowledge, we demonstrate a significant association of the IL-1A-889 functional polymorphism with CAD.

Prevalence of hepatitis B virus in the Kurram Agency, Pakistan: A 5-year observational study in a war-affected region

BACKGROUND Hepatitis B is an inflammatory liver illness caused by the hepatitis B virus. The exact magnitude and extent of the hepatitis B virus in Pakistan is still unknown, but at least 10 million people are estimated to be infected with chronic hepatitis B virus. OBJECTIVES The objective of this study was to determine the seroprevalence of hepatitis B surface antigen (HBsAg) in the war-affected area since 2009. STUDY DESIGN Observational study. RESULTS A total of 4922 healthy subjects were tested for the detection of HBsAg during 2009-2013, and 14.95% subjects were found to be reactive for HBsAg. The highest seroprevalence (26.0%) of HBsAg was found in those individuals who were less than 35 years of age. Male subjects were more affected (16.6%) than females. The seroprevalence of HBsAg was significantly associated with <35 years of age and male gender (P<0.005 for both). Moreover, an increasing trend over 5 years was observed, as 8.6% subjects were positive for HBsAg in 2009 and subsequently 10.4% in 2010, 14.6% in 2011, 18.9% in 2012, and 21.7% were reactive for HBsAg in 2013. CONCLUSION This study concluded that HBsAg was more prevalent in the war-affected region. The prevalence rate was increasing with time as the highest rate was found in 2013. Present observations will help to provide the background for awareness and bring the increasing levels of hepatitis B to the attention of health professionals and government authorities in order to increase the capacity of the health systems in such troubled areas.

Association of Serum Uric Acid with Family History of Coronary Artery Disease in a Pakistani Population.

To the Editor, Coronary artery disease (CAD) or coronary atherosclerosis is a major cause of death and disability around the world. Exploring the complex nature of CAD is not an easy and simple task. In fact, CAD is a heritable trait and has remained the focus of advanced research for the last few decades. Multiple factors including hyperlipidemia, hypertension, diabetes mellitus, family history, and genetics are all assumed to be involved in the pathogenesis of CAD (1). Understanding all of the established and novel emerging risk factors by clinicians is critical in order to prevent cardiovascular disabilities. Despite the clear involvement of established risk factors, clinical studies have shown that serum uric acid (SUA) is associated with hypertension or other cardiovascular diseases (2). The pathological role of elevated SUA in CAD either independently or in association with other cardiovascular disease risk factors has been the topic of debate. Recently, a Mendelian-randomized study showed a positive relationship between body mass index, hyperuricemia and elevated SUA (3). However, the increased serum uric acid and its association with a family history of CAD has not been published. Therefore, the aim of the present study was to investigate the association of SUA with a family history of CAD. The present study protocol is adherent to the Helsinki Declaration of 1975 as revised in 1997 and was approved by the Ethical Committee, COMSATS Institute of Information Technology, Islamabad. A total of 330 subjects consisting of 110 patients with CAD (group 1), 110 unrelated healthy subjects without a family history of CAD (group 2), and 110 healthy controls with a family history of CAD in at least three generations (group 3) were analysed. Biochemical analyses of lipid profile, and SUA were determined by using the AMP Diagnostics kits (Austria) with the help of a Vitalab Selectra E chemistry analyzer (Netherlands). The inclusion criterion was applied for both the patients and controls in this study. Study subjects were evaluated by physical and clinical history, ECG, and coronary angiography. The results were evaluated by using Chi-square and One-Way ANOVA. The basic and clinical variables are listed in Table 1. Lipid biomarkers were significantly associated with CAD in the study population. SUA levels were 5.7±1.20 in CAD patients, 5.5±1.0 in controls with CAD family history and 3.8±0.65 in healthy subjects with no family history. SUA was significantly increased in CAD patients compared with controls with no family history (5.7±1.20 mg/dL vs 3.8±0.65 mg/dL, p<0.0001). The comparison of SUA was not significantly different between CAD patients and controls with a positive family history of CAD (5.7±1.20 vs 5.5±1.0, p=0.235). Beside, a significant difference was observed between the subjects with a family history of CAD and those with no such history (5.5±1.0 vs 3.8±0.65, p<0.0001). TABLE 1. Baseline and clinical characteristics of study population (n=330) The pathological role of elevated SUA in CAD has been suggested by several possible mechanisms (2). The underlying atherosclerotic mechanism may involve abnormal xanthine oxidase, oxidative stress, and the formation of free radicals causing a deleterious effect on endothelial cell function. Increased SUA stimulates inflammation and promotes platelet adhesiveness (4). However, recent data regarding the association of SUA with CAD in a dependent or independent manner is not very clear (5). In our study, for the first time, we have established a significant association of SUA with a family history of CAD, and this should be considered a risk factor for CAD in this high risk population.