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Genetic variations in a disintegrin and metalloprotease 12 (ADAM12) gene may contribute to develop Osteoarthritis (OA) that is characterized by cartilage matrix degradation and osteophytes formation. Therefore, the aim of present study was to analyze the association between the ADAM12 gene variants and knee OA predisposition. Tetra-primers ARMS-PCR was employed, to genotype the ADAM12 gene polymorphisms (rs1044122 and rs1871054) in 400 knee OA patients and equal number of age-matched controls. The association between ADAM12 gene variants and OA susceptibility was estimated using the Chi-square, logistic regression, haplotypes and linkage analyses. A significant association of rs1044122 (genotype: χ

Objective: To assess the relationship of tumour necrosis factor-alpha with obstructive sleep apnoea and its severity in Pakistani population. Method: The cross-sectional study was conducted at the Sleep Laboratory of Dow University Hospital, Karachi, from December, 2018, to March, 2020, and comprised patients of either gender having symptoms of snoring, witnessed apnoea or daytime sleepiness. They were divided into four groups on the basis of obstructive sleep apnoea status. Those without obstructive sleep apnoea were in Group A, mild status in Group B, moderate in Group C and severe obstructive sleep apnoea in Group D. Polysomnography was done to confirm obstructive sleep apnoea status and to categorise the subjects using apnoea-hypopnea index, while enzyme-linked immunosorbent assay was used to assess their tumour necrosis factor alpha levels.. Data was analysed using SPSS 20. Results: Of the 150 subjects, 94(63%) were males. The overall mean age was 49.68±12.14 years. There were 50(33.33%) subjects in Group A, 19(12.66%) Group B, 23(15.33%) Group C and 58(38.66%) in Group D. Mean tumour necrosis factor-alpha level was 3.88±1.65pg/mL in Group A, 9.97±4.33pg/mL in Group B, 12.65±4.46pg/mL in Group C and 12.83±4.33pg/mL in Group D. Mean tumour necrosis factor-alpha levels had significant association with the severity of obstructive sleep apnoea (p

Different pesticide chemicals are used to enhance crop yield by protecting from pests. Organophosphate (OPs) and Pyrethroid (PYR) are used in fields of Sanghar, Sindh Pakistan. PON1 an antioxidant enzyme implicated in OPs detoxification may predispose by OPs chronic exposure. This study was conducted to evaluate the toxic potential of active pesticide chemicals at cellular and genetic levels. To examine toxic potential, locally consumed pesticide n = 2 and reference pesticide compounds organophosphate (OPs): Chloropyrifos, Malathion and Pyrethroid (PYR): Cyprmethrin, Cyhalothrin n = 4 were tested against NIH 3T3 cells using MTS assay. Local consumer pesticides demonstrated relevance for half-maximum inhibitory concentration (IC50) 0.00035 mg/mL with selected compound. Malathion IC50 exhibited the highest cytotoxicity among four compounds at 0.0005 mg/mL. On genotoxicity analysis in exposed subjects n = 100 genotypes and alleles n = 200 exhibited significant differences in genotypic and allelic frequencies of pesticide exposed subjects and controls n = 150 (X2 = 22.9, p = 0.001). Screening of genotypes were performed by PCR- RFLP. Statistical assessment carried out using online software and tools. Results suggested that higher heterozygous genotype A/G (74%) may confer low PON1 metabolic activity towards pesticides in exposed subjects. Findings could be helpful to establish health plans by avoiding toxic chemicals that harming exposed population.

Objective: To investigate the correlation of Adenine-1012-Guanine (rs4516035) promoter region polymorphism of vitamin-D receptor gene with serum levels of omentin-1, vitamin-D and vitamin-D receptor protein in patients with coronary artery disease. Method: The case-control study was conducted January to June 2020 at the cardiac unit of Civil hospital Karachi (CHK), and comprised coronary artery disease patients and controls. The tetra-primer amplification refractory mutation system polymerase chain reaction method was used to genotype Adenine-1210Guanine polymorphism in the vitamin D receptor gene. Serum levels of omentin-1, vitamin-D, and vitamin-D receptor protein were measured in both the groups using an enzyme-linked immunosorbent assay. Data was analysed using SPSS 17. Results: Of the 1,000 subjects, there were 500(50%) cases; males 306(61.2%) and 194(38.8%) females with overall mean age of 51.08±9.55 years. The remaining 500(50%) were controls; 290(58%) males and 210(42%) females with overall mean age of 50.9±10.78 years. The mutant Guanine allele was more prevalent in controls 261(52.2%), and had a non-significant correlation with coronary artery disease (p=0.45). Among the cases, the wild Adenine-Adenine genotype had a higher prevalence 402(80.4%) and had a significant correlation with coronary artery disease (p

Epilepsy is one of the most common neurological disorders with the incidence rate higher in developing states. It is a multifactorial ailment in which genetic diversity along with other factors plays an important role. The objective of this study was to assess the involvement of different risk factors including single nucleotide polymorphisms (SNPs) present in GABRA1 (rs2279020) and GABRG2 (rs211037) genes with the susceptibility to epilepsy in the targeted population. Blood samples of 180 subjects were taken and genotyped through tetra-primer amplification refractory mutation system-polymerase chain reaction technique. The obtained demographic and genotypic data were analyzed through different statistical tools including χ

BACKGROUND Coronary artery disease (CAD) is a leading cause of mortality in Pakistan and also worldwide. Vitamin D receptor (VDR) regulates the transcription of many genes and has a significant impact on inflammation and the morphology of cardiac cells. Genetic variation in the VDR gene such as the TaqI polymorphism (rs731236) may have an impact that causes adverse effects. Accordingly, it is important to determine possible association of the TaqI polymorphism (rs731236) with CAD. METHODS The study included blood samples from 1016 subjects: 516 from CAD patients and 500 from age- and gender-matched controls. Genomic DNA was extracted by standard salting out method. Targeted variation was amplified by an allele-specific polymerase chain reaction (PCR). PCR products were examined and genotyped on agarose gel electrophoresis represented by an amplified product size of 148 bp followed by Sanger sequencing to validate variations. RESULTS Serum vitamin levels, as observed using enzyme-linked immunosorbent assay, were found to be insufficient in both CAD patients (20.52 ± 0.06 ng/ml) and controls (21.6981 ± 0.05 ng/ml). The TaqI polymorphism (rs731236) T>C was found to be significantly associated with CAD (p

Background The genetic polymorphisms (rs708035, rs3844283) of Interleukin-1 receptor associated kinases 2 (IRAK2) is involved in the NFκB regulatory pathway. The frequencies of IRAK2 gene are unknown in Pakistani population. Therefore, the study was designed to examine the association of targeted single nucleotide polymorphism(s) in IRAK2 gene of RA patients. Methodology The study participants were selected by ACR/EULAR 2010 standards. After ethical approval, the blood samples of patients and healthy controls were collected for the extraction of DNA followed by the amplification of targeted polymorphism(s) via Tetra-primer Amplification Refractory Mutation System (T-ARMS PCR). Desired products were observed via agarose gel electrophoresis. Results The allele frequency of wild type A and C is frequent among patients and mutant T and G is frequent among controls. The rs708035 showed significant protective association while rs3844283 was found to be associated with risk of RA. Genetic model associations were applied to determine the role of genotypes. In combination analyses of alleles revealed AC haplotype was found to be associated with risk and TG provide protection against RA. Moreover, targeted SNPs were found to be in 61% Linkage Disequilibrium among the targeted population. Conclusions Current study revealed the protective and risk association of targeted SNPs (rs708035, rs3844283). Study might be beneficial as it provides baseline data regarding targeted SNPs and their role in the disease progression. This could be served as potential biomarker for diagnostic purpose and effectively utilized in precision medicine approach.

The study was designed to investigate the association between obesity and the risk of knee osteoarthritis, recruiting 400 knee osteoarthritis patients and an equal number of controls. After the informed consent, diagnosed patients from Jinnah Post Graduate Medical Centre, Karachi were included as "cases". Age-matched individuals without the disease were included as "controls". Sociodemographic data were taken from each participant. Characteristics were compared by odds ratio and chi-square using SPSS 20 software. Obesity (OR 3.29; 95% CI 2.40-4.51), female gender (OR 2.87; 95% CI 1.94-4.25) and family history (OR 3.61; 95% CI 2.69-4.85) were found to be significantly associated with osteoarthritis (p0.001). Highest OR was found in case of stair climbing10 flights/d (OR 6.08; 95% CI 4.16-8.89; p0.001), whereas heavy lifting (25 kg/d for4 hr) was observed as another major factor with OR of 5.24 (95% CI 3.54-7.75; p0.001) that elevates the risk. The study concluded that obesity is significantly associated with osteoarthritis and obese individuals (BMI25 kg/m2) are at high risk of disease development. Furthermore, family history, prolonged standing (2 h/d for1 yr), heavy lifting (25 kg/d for4 hr), stair climbing (10 flights/d) and sitting on the floor (5 h/d) might also be associated with knee osteoarthritis.

This study was conducted to find the association between leptin and adiposity indices. Secondly, to identify optimal threshold of various anthropometric indices for obesity, as assessed by 75th percentile of leptin levels, within a clinic sample of non-diabetic and diabetic Pakistani adults. Fasting serum leptin levels were compared with anthropometric markers of obesity in 164 diabetic and non-diabetic subjects (90 male, 74 female), aged 35 to 65 years. Obesity was defined by body mass index (BMI) of 25 kg/m2 in either sex. The cutoff point of leptin was taken as the 75th percentile in non-obese subjects. Diagnostic accuracy for detecting excess fatness was evaluated through receiver operating characteristics (ROC) analyses with leptin taken as reference test against anthropometric indices as test variables. The 75th percentile of leptin in male and female was 7.0ng/mL and 17.9ng/mL, respectively. Leptin levels were significantly higher in females (p0.001) and had strong positive correlation (p0.001) with most anthropometric indices of obesity in both sexes; hip circumference (HC) being most prominent among these. Largest area under ROC curve (AUC) was between WC and leptin (AUC=0.844; CI=0.764, 0.925) in males and BMI and leptin (AUC=0.832; CI=0.740, 0.923) in females. The optimum thresholds for obesity indices in our study were: BMI, WC and HC as 25 kg/m2, 96.25cm, 99.25cm for males; 27 kg/m

Chemokine CC receptor type 5 (CCR5) is a cell surface receptor that has high affinity for chemotropic cytokines called chemokines. The CCR5 gene contains a 32 base pairs (bp) deletion (CCR5Δ32). This deletion may result in a malformed and nonfunctional receptor, reported to be responsible for the development and dissemination of different cancers. CCR5Δ32 exists in two allelic forms i.e. deletion (D) and wild type (WT). This study aims to detect the role of CCR5Δ32 in breast cancer development. Blood samples were collected from breast cancer patients (330) and controls of same gender (306). Along with this histopathologically diagnosed malignant tissue samples were also excised from breast lesions of 100 patients. Genetic variations within the blood and tissue samples were examined by PCR then observed through gel electrophoresis and confirmed by direct DNA sequencing. Obtained DNA sequences were aligned and analyzed by MEGA6 software. Genotypic and association analyses were done by SPSS software version 17.0. Deletion of 32 bp in CCR5 gene has been analyzed. Genotypic variations of CCR5Δ32 are; homozygous wild type (WT/WT), heterozygous deletion (WT/D) and homozygous deletion (D/D). Statistical analyses of CCR5Δ32 data revealed that WT/D was significantly higher in blood samples of breast cancer patients (7.27% (24/330)) as compare to controls (1.30% (4/306)). In tumor tissue samples WT/WT being the most frequent genotype (99.00% (99/100)) with 1.00 (1/100) of D/D which suggested that it may be acquired. Hence, association analysis showed that CCR5Δ32 is positively associated with breast cancer in Pakistan (p

To explore the usefulness of neck circumference as a supplemental tool for diagnosing metabolic syndrome while identifying its cut-off values.This case-control study was conducted at Dr. Essa's Laboratory and Diagnostic Centre, Karachi, from December 2014 to April 2015, and comprised subjects with and without metabolic syndrome aged between 35 and 65 years regardless of their diabetic status. Evaluation was done for metabolic syndrome by measuring anthropometric, clinical and biochemical parameters according to the criteria proposed by the International Diabetes Federation. Variables in both cases and controls were correlated with neck circumference and its cut-off values were determined for diagnosing metabolic syndrome. SPSS 20 was used for statistical analysis.Of the 215 subjects enrolled, 164(76.28%) were selected. Of them, 83(50.61%) were cases and 81(49.39%) were controls. Moreover, 90(55%) of them were men and 74(45%) were women. The overall mean age was 51.15±10.36 years (range: 35 to 65 years).The mean neck circumference was 36.13±2.14 cm and 31.59±1.18 cm in normal-weight men and women, respectively, compared with 40.0±2.13 cm and 35.75 ± 2.74 cm among obese men and women. The neck circumference correlated best with waist circumference in men (p=0.001) and with body surface area in women (p=0.001). The area under the curve of neck circumference for metabolic syndrome was 0.760 for men (p0.001) and 0.631 for women (p0.05). Optimal neck circumference cut-off points to determine metabolic syndrome were38 cm for men and34 cm for women. The odds ratio for metabolic syndrome was 12.44 (95% confidence interval: 4.13-37.41) among male cases and controls compared to 3.34 (1.26-8.80) among women.Neck circumference strongly correlated with adiposity indices and had a definite cut-off point. It can therefore be used as a useful adjunct for clinical screening of metabolic syndrome.

The identification of chromosomes for routine cytogenetic analysis is based on quality of metaphases and good banding pattern. Fresh slides of human lymphocytes have been shown to produce good bands for the identification of chromosomes morphology. G-bands by Trypsin using Giemsa (GTG) banding of aged slides is generally considered hard to get desired band pattern of chromosomes persistently. The current study is focused on GTG banding of aged slides. A total of 340 subjects including 290 primary infertile and 50 fertile were selected. The blood samples were drawn aseptically for cytogenetic analysis. Lymphocytes were cultured and GTG banding was done on 1440 glass slides. Giemsa trypsin banding of aged slides were done by adjusting average trypsin time for each month according to the slide age and metaphase concentration. Correlation analyses showed a significant and positive correlation between slide ageing and trypsin pre-treatment time. The results of this study suggest that, the fresh and aged human lymphocyte metaphases are equally usable for GTG banding.