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Abstract Background Psoriasis is a chronic, autoimmune, papulosquamous skin disorder, characterized by the formation of drop-like papules and silvery-white plaques surrounded by reddened or inflamed skin, existing predominantly on the scalp, knees and elbows. The characteristic inflammation and hyperproliferation of keratinocytes in psoriasis is regulated by progranulin (PGRN), which suppresses the expression and release of inflammatory cytokines, such as TNF-α. Methodology In this study mutation analysis of the PGRN gene was performed by extracting the genomic DNA from blood samples of 171 diagnosed psoriasis patients and controls through standard salting-out method, followed by amplification and sequencing of the targeted region of exon 5–7 of PGRN gene. Results Three single nucleotide polymorphisms, rs25646, rs850713 and a novel point mutation 805A/G were identified in the PGRN gene with significant association with the disease. The variant alleles of the polymorphisms were significantly distributed among cases and controls, and statistical analysis suggested that the mutant genotypes conferred a higher risk of psoriasis development and progression. Multi-SNP haplotype analysis indicated that the CAA (OR = 8.085, 95% CI = 5.16–12.66) and the CAG (OR = 3.204, 95% CI = 1.97–5.21) haplotypes were significantly associated with psoriasis pathogenesis. Conclusions These findings demonstrate that polymorphisms in PGRN might act as potential molecular targets for early diagnosis of psoriasis in susceptible individuals.

Objective: To assess the link between tumour necrosis factor-alpha -308 guanine/adenine polymorphism and tumour necrosis factor-alpha plasma levels in relation to obstructive sleep apnoea. Method: The cross-sectional study was conducted from December 2018 to March 2021 at the sleep clinic of Dow University Hospital, Karachi, on obstructive sleep apnoea patients and healthy controls. Epworth Sleep Scale score was used to determine daytime sleepiness, while full-night polysomnography was carried out for obstructive sleep apnoea confirmation and categorisation according to severity. Blood sample collection was followed by deoxyribonucleic acid extraction and plasma tumour necrosis factor-alpha measurement using enzyme-linked immunosorbent assay. Genotype distribution and allelic frequency were assessed. Data was analysed using SPSS 20. Results: Out of the 225 subjects, with a mean age of 47.68±9.88 years, 132 (58.7%) were males, and 93 (41.3%) were females. Among them, 150 (66.7%) were patients, and 75 (33.3%) were controls. Heterozygous tumour necrosis factor-alpha -308 guanine/adenine genotypes were significantly higher among the patients (p

Rheumatoid arthritis (RA) is an autoimmune disorder with unknown aetiology. Patients suffering from RA face persistent pain due to joint inflammation, and tissue destruction. Behavioural phenotyping is an approach to target the role of different behavioural traits associated with disease progression. The study aimed to assess behavioural patterns associated with decreased muscle strength in the adjuvant-induced rheumatoid arthritis animal model. The study was conducted on male Albino Wister rats (n = 30) [Control, Vehicle, and Disease groups]. After taking ethical approvals RA was induced by complete Freund's adjuvant (CFA) intradermally base of tail. The weight of animals, macroscopic analysis of inflammatory signs, and arthritic scores were measured weekly. Grip strength, ganglia-based movement, cataleptic activity, and motor-coordination-related behaviours were assessed among the groups. Radiographs and spleen index assay were performed followed by data analysis using one-way and two-way ANOVA (Analysis of Variance). A significant decrease in weight and an increase in arthritic scores among the diseased group was observed. Behavioural analyses confirmed that diseased animals had significantly decreased grip strength and increased cataleptic activity with less motor coordination. Radiographic images and spleen index assay confirmed the pattern of RA. Therefore, it can be suggested that the development of the disease animal model is an effective approach to identifying the disease progression and associated behavioural changes. Moreover, this prepared laboratory animal model may be utilised for pathway analyses to understand the key role of immune regulators and genetic insight into molecular pathways associated with acute and chronic phases of RA.

Abstract 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 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

BackgroundThe 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.MethodologyThe 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.ResultsThe 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.ConclusionsCurrent 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.

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

Taxonomic characterization of bacterial 16S rRNA gene has become an accepted norm for inference of complex microbial communities and ecological shifts in gut. Recently, 16S rRNA gene sequences has emerged as a taxonomic marker to investigate crosstalk between gut bacteria and host via variations in feed. Here, broiler chickens (n = 240) were subjected to dietary modulation approach to substitute antibiotics with natural plant-derived products. Five experimental groups were control, antibiotic growth promoter, organic acids, phytogenic feed additives and combination (organic acids + phytogenic feed additives). Bacterial DNA was extracted and 16S rRNA gene was amplified through polymerase chain reaction using universal primers followed by observation through gel electrophoresis and then, confirmation by direct DNA sequencing. The sequence data was analyzed for homology search, multiple sequence alignment, and construction of phylogenetic clades using software BLASTn, CLUSTAL W and MEGA 7.0. respectively. Based on the phenetic relationship, bacterial phylotypes from organic acid and phytogenic feed additive groups belongs to Enterococcus faecium and Enterococcus faecalis in both cecum and ileum. The bacterial isolate obtained from antibiotic feeding group characterized as Escherichia coli. Furthermore, sequences of probiotic bacteria (Enterococcus faecalis) were retrieved from online repositories (GenBank and RDP) and aligned with indigenously isolated strains of this study. Comparative phylogenetic analysis revealed high sequence similarities (99%) of Enterococcus faecalis KIBGE AB-6 and Enterococcus faecalis KIBGE AB5 with Enterococcus faecalis ATCC 19433, and Enterococcus faecalis DD28 respectively. Hence, under the appropriate conditions of current investigation, these bacterial strains might serve as potential candidates for probiotics in animals. To the best of our knowledge, this is the first study in Pakistan depicting taxonomic classification of chicken gut bacterial diversity from cecal and ileal samples linked with dietary modulation. Keywords: 16S rRNA gene, Broiler chicken, Gut microbiota, Probiotic bacteria, Taxonomy

Background & Objective: Coronary artery disease (CAD) is a most important cause of morbidity and mortality worldwide as well as in Pakistan. Recent studies have shown that the combination of obesity, insulin resistance and fluctuation in circulating adipocytokines levels is associated with the pathogenesis of coronary artery disease. Omentin-1 is recently found adipocytokine that is highly expressed in visceral adipose tissue. It has anti- inflammatory properties and is negatively correlated with ischemic heart disease. Therefore, this study was designed to investigate the relationship between omentin-1 Val109Asp polymorphism and CAD in Pakistani population. Methods: A total of 350 subjects were included in the study. Two hundred fifty were diagnosed with coronary artery disease while 100 served as healthy controls. PCR-RFLP was performed at Dr. A Q. Khan Institute of Biotechnology (KIBGE) to analyze Val109Asp polymorphism. In this, case control study SPSS software version 16 (Chicago, IL, USA) was used for data analysis. Continuous variables and categorical variables were presented as mean±SD or in percentage. Independent sample test and chi-square test was performed to compare the differences in means between cases and controls. Genotype distribution was analyzed by chi-square test and results were presented as percentage and frequency. Multivarible regression analysis indicated that Val109Asp SNP might be an independent risk factor for CAD susceptibility after adjustment for some well- known CAD risk factors including age, gender, body mass index, smoking, hypertension, diabetes mellitus and lipid abnormalities. There was estimation of odd ratios (OR) and 95% confidence intervals (CIs) to determine the correlation between genotypes and the risk of CAD. (p> 0.05). Genotype frequencies were compared by Chi-square test. Results: There was prevalence of Omentin-1 Val109Asp polymorphism in both case and control groups. However, Val/Asp (heterozygous mutant) genotype was detected more frequently in patients with CAD, OR(95%)=1.921; CI=1.173-3.1469 in comparison of Asp/Asp and Val/Val genotypes. Conclusion: Individuals having Val/Asp heterozygous gemotype of omentin-1 gene polymorphism are at more risk of developing CAD in Pakistani population, further studies are required in different populations and ethnicities to confirm our findings. [ABSTRACT FROM AUTHOR]

Background & Objectives: Triple negative and triple positive breast cancer have adverse effects than other types of breast cancer. However, triple negative has poor prognosis with short survival as compared with triple positive breast cancer. Good prognosis is one of the key factors for successful treatment trial. This study aimed to find out the association of sociodemographic and reproductive features like parity, menopause, number of child bearing as risk factors in the development and prognosis of triple negative and triple positive breast cancer., Methods: This study is a part of an ongoing project which is being conducted in Karachi from 2013 to 2020. Informed consent from triple negative breast cancer (n=134) and triple positive breast cancer (n=87) patients were taken prior to their recruitment into the study. Demographic, anthropometric, reproductive and disease history of patients were recorded. Means, frequency distribution, data classification and association analyses were done by SPSS version 17.0., Results: Statistical analyses revealed that delayed first child bearing age and lower number of children are associated with the development of triple negative breast cancer. However, no significant effect of these parameters has been observed on the outcomes of triple positive breast cancer., Conclusions: Reproductive factors have more pathological implications than sociodemographic factors in both triple positive and triple negative breast cancer development. These findings might prove to be beneficial for effective and better breast cancer management., Competing Interests: Conflict of Interest: None.

Objective: The myokine interleukin-15 (IL-15) is capable of modifying the metabolism of both skeletal and adipose tissue. This study compares the change in serum levels of IL-15 in obese and non-obese after a single session of submaximal exercise., Methods: A cross-sectional study was carried out at Jinnah Medical and Dental College, Karachi, during Aug-Dec 2015, comprising of 133 medical students (aged 17-24 years). Cardiorespiratory fitness was evaluated by Queen's College Step Test. Blood was obtained both before and just after exercise and serum levels of IL-15 determined by enzyme-linked immunosorbent assay., Results: Mean serum level of IL-15 was 3.64±1.59 pg/mL. Higher levels of IL-15 were seen in lean subjects compared to overweight/obese, both before and after three minutes of exercise (all P trend <.001). The percent increase in IL-15 upon exercise was 12.7% higher in lean. Significant negative association was seen between interleukin-15 and adiposity, especially visceral fat ( r = -.288, p =.001)., Conclusion: Interleukin-15 correlates negatively with adiposity indices, especially visceral fat. With the proven benefit of IL-15 in terms of adipose tissue stores and skeletal muscle mitochondrial biogenesis, endurance exercises, even of short duration, may possess therapeutic potential towards producing a healthier body.

Objective: Biotransformation of xenobiotics are critical for their metabolism and removal from the body which is carried out by xenobiotic metabolizing enzymes. Individuals carrying variants of genes that encode these enzymes have an altered ability to metabolize xenobiotics which may lead to an increased risk of acute lymphoblastic leukemia. The current study aimed to investigate the impact of GSTM1 and GSTT1 gene deletions in causing predisposition to adult ALL., Methods: The current case-control study involved 62 adult ALL patients and 62 age and gender matched healthy controls. Whole blood samples processed with standard phenol chloroform protocol for DNA isolation were genotyped using multiplex PCR approach for simultaneous identification of GSTM1 and GSTT1 deletions. The genotype frequency obtained for patients was compared to controls using odds ratio and chi-square., Results: The null genotype frequency of GSTM1 and GSTT1 in a group of adult ALL patients from Pakistan were 47% and 11% respectively. Deletion of GSTM1 and GSTT1 did not show statistically significant association with adult ALL ( p =0.86 and p =0.35 respectively). The combined GSTM1/GSTT1 deletion was observed in 2% patients and was not significantly associated with ALL in adults ( p =0.85)., Conclusions: The results reveal that homozygous null polymorphism of GSTM1 and GSTT1 genes does not influence ALL susceptibility among adult patients. Cancer susceptibility associated with GST polymorphism varies with ethnic and geographic differences. Therefore, further investigation on different populations is needed to understand the role of these genetic variations in modifying adult ALL risk.

Background & Objective: Catalase (CAT) is an important endogenous antioxidant enzyme that detoxifies H 2 O 2 into water and oxygen, consequently limiting the deleterious effects of reactive oxygen species. It has suggested that CAT-21A/T (rs7943316) OMIM: 115500 gene promoter polymorphism is predominantly associated with different human disorders such as hypertension, cancers, diabetes, nephropathy, and other diseases accompanied by oxidative stress. This study was designed to investigate the prevalence of mutant T allele frequency in healthy individuals., Methods: The study group consisted of 110 healthy individuals were enrolled from Baqai Institute of Diabetology and Endocrinology (BIDE), Karachi, Pakistan, during the period of April 2010 to May 2013. DNA was isolated from leukocytes. Genotyping of CAT-21A/T (rs7943316) gene promoter polymorphism was carried out using thermal cycler followed by RFLP. Blast N analysis was performed for the confirmation of gene sequences., Results: In CAT-21A/T (rs7943316) gene promoter polymorphism, wild type genotype (AA) was observed in 18.26% and alterered genotype (AT/TT) found in 81.74% cases., Conclusions: Data demonstrates that frequency and distribution of mutant T allele was more prevalent as compared to wild type A allele in the study group., Competing Interests: Declaration of Interest: None.

Background & Objective: Coronary artery disease (CAD) is a most important cause of morbidity and mortality worldwide as well as in Pakistan. Recent studies have shown that the combination of obesity, insulin resistance and fluctuation in circulating adipocytokines levels is associated with the pathogenesis of coronary artery disease. Omentin-1 is recently found adipocytokine that is highly expressed in visceral adipose tissue. It has anti- inflammatory properties and is negatively correlated with ischemic heart disease. Therefore, this study was designed to investigate the relationship between omentin-1 Val109Asp polymorphism and CAD in Pakistani population., Methods: A total of 350 subjects were included in the study. Two hundred fifty were diagnosed with coronary artery disease while 100 served as healthy controls. PCR-RFLP was performed at Dr. A Q. Khan Institute of Biotechnology (KIBGE) to analyze Val109Asp polymorphism. In this, case control study SPSS software version 16 (Chicago, IL, USA) was used for data analysis. Continuous variables and categorical variables were presented as mean±SD or in percentage. Independent sample test and chi-square test was performed to compare the differences in means between cases and controls. Genotype distribution was analyzed by chi-square test and results were presented as percentage and frequency. Multivarible regression analysis indicated that Val109Asp SNP might be an independent risk factor for CAD susceptibility after adjustment for some well- known CAD risk factors including age, gender, body mass index, smoking, hypertension, diabetes mellitus and lipid abnormalities. There was estimation of odd ratios (OR) and 95% confidence intervals (CIs) to determine the correlation between genotypes and the risk of CAD. (p> 0.05). Genotype frequencies were compared by Chi-square test., Results: There was prevalence of Omentin-1 Val109Asp polymorphism in both case and control groups. However, Val/Asp (heterozygous mutant) genotype was detected more frequently in patients with CAD, OR(95%)=1.921; CI=1.173-3.1469 in comparison of Asp/Asp and Val/Val genotypes., Conclusion: Individuals having Val/Asp heterozygous gemotype of omentin-1 gene polymorphism are at more risk of developing CAD in Pakistani population, further studies are required in different populations and ethnicities to confirm our findings.

Objective: This study investigated the relationship between cardiorespiratory fitness (CRF) and adiposity in young adults., Methods: Data was collected from 133 students of a medical college of Pakistan. The study was conducted on young adults, aged 17-24 years, recruited from Jinnah Medical & Dental College, Karachi, between Aug-Dec, 2015. Queen's College Step Test was conducted to measure CRF and maximal oxygen uptake (VO 2max ) evaluated. Anthropometric measurements (body mass index, body fat, visceral fat, waist circumference) were taken to assess adiposity. Associations of VO 2max and adiposity were analyzed., Results: The prevalence of overweight/obesity was 44% overall. The VO 2max (ml/kg/min) of males and females was 55.41±9.45 and 39.91±3.14, respectively, the gender difference being highly significant ( p <0.001). Quartiles of VO 2max showed strong inverse relationship between adiposity and VO 2max, obese individuals having low VO 2max (1 st quartile) and normal weight individuals having high VO 2max (4 th quartile). VO2max correlated greatest with body fat in males (r = -0.600; p<0.001), and waist circumference in females (r = -0.319; p=0.004)., Conclusion: The results indicate low CRF in young females and a strong inverse relationship between fitness levels and adiposity in young adults of both genders. Improving these parameters in our young population may prevent development of chronic non-communicable disease in later life., Competing Interests: Declaration of interest: The authors declare no conflict of interest.