Genetic Polymorphism and serum levels of Insulin like Growth Factor-1 (Igf-1) in patients of rheumatoid arthritis.

Objectives: Insulin like growth factor-1(IGF-1), is a modulator of immunity and inflammation, it promotes the anabolic role of growth hormone (GH) on bone and skeletal tissue. Genetic polymorphism in IGF-1 gene is reported to affect the transcriptional efficiency affecting its serum level. In this study we aim: 1) To study the presence of 192bp polymorphism of IGF-1 gene in patients of rheumatoid arthritis (RA), 2) To study the association of 192 bp polymorphism of IGF-1 gene with serum IGF-1 levels and disease severity in patients of RA. Methods: A cross-sectional study was carried out at University of Health Sciences (UHS), Lahore. Diagnosed RA cases who fulfilled the American College of Rheumatology (ACR) criteria were recruited from Fatima Memorial Hospital (FMH) and Behbud Rheumatology Clinics, Lahore during 2018-2019. Serum IGF-1 levels were determined by ELISA in blood samples of 200 RA patients and 200 healthy individuals. DNA was extracted and genetic polymorphism was determined. Results: The serum IGF-1 level in RA group was significantly lower compared to healthy group. Our study shows presence of 192bp allele of IGF-1in 77% of the studied population. Carriers of 192bp allele of IGF-1 had a significantly higher serum level of IGF-1 as compared to non-carriers in the RA patients. Rheumatoid factor (RF) positive patients had a higher number of 192bp carriers in comparison to RF negative patients. Significant difference was also seen in severity of disease between carrier and non-carriers of 192bp allele with the disease being more severe in male carriers. Conclusions: There is an association of IGF-1gene polymorphism with variation in serum IGF-1 levels and severity of RA.

Copy number variations and protein expression of Cyclin E1 and Epithelial cell transforming sequence 2 genes predict the chemotherapeutic response in patients with serous ovarian carcinoma.

Background & Objectives: Serous ovarian carcinoma (SOC) is characterized by extreme genomic instability, chromosomal rearrangements and copy number variations (CNVs) leading to the development of early metastasis and chemo-resistance. The present study was designed to observe the role of CNVs of Cyclin E1 (CCNE1) and Epithelial cell transforming sequence- 2 (ECT2) genes and their encoded proteins in predicting the chemotherapeutic response in SOC patients. Methods: This observational analytical study was conducted at University of Health Sciences, Lahore, Pakistan from December 2019 till June 2022.The study included twenty-five SOC patients with resectable ovarian tumors and twenty-five control subjects. The patients were followed-up for six months for their response to chemotherapy. The CNVs in CCNE1 and ECT-2 genes were determined by real time PCR while serum levels of encoded proteins were determined in controls and cases, before and after six months of treatment, through ELISA. The response to chemotherapy was categorized as sensitive or resistant based on serum CA-125 levels and radiological scans. Results: The copy number variations in CCNE1 and ECT2 genes showed association with the clinic-pathological characteristics and chemotherapy response. Statistically significant difference was found between the mean pre-chemotherapy protein levels of CCNE1 in cases than controls (p-value <0.001) and between the mean pre and post-chemotherapy protein levels of CCNE1 and ECT2 (p-value <0.001) in SOC patients. Conclusion: The copy number variations of CCNE1 and ECT2 genes and their protein expression are positively associated with chemotherapeutic response in SOC patients.

Moringa oleifera alleviated oxidative stress, physiological and molecular disruption induced by acute thermal stress in grass carp, Ctenopharyngodon idella.

The present study was designed to investigate the protective effect of dietary Moringa oleifera leaf meal (MLM) supplementation against high temperature-induced stress in grass carp (Ctenopharyngodon idella). A total of 180 apparent healthy juvenile grass carp (15.48 g ± 0.054) were divided into three groups in triplicate (20 fish in each replicate). Fish were fed with diets containing 0, 1, and 5% MLM for 60 days and then subjected to a high-temperature challenge for 48 h (32-33 °C). The results revealed that feeding fish with 1 and 5% MLM resulted in a significant increase in weight gain and specific growth rate compared to the control. In addition, feed conversion ratio was significantly reduced in groups fed with MLM. No significant difference was reported in the serum cortisol level among different experimental groups before heat stress while serum glucose level significantly decreased in fish fed with 5% MLM. Serum alanine transaminase, aspartate transaminase, and alkaline phosphatase significantly decreased in fish fed with 1 and 5% MLM before and after heat stress. Hepatic lipid peroxidation significantly decreased in fish fed with MLM for 60 days. A non-significant increase in hepatic reduced glutathione level was reported in fish fed with 1 and 5% MLM before heat stress. Catalase and superoxide dismutase activities increased significantly in the liver of fish fed with 5% MLM. No significant change was observed in the expression profile of heat shock protein (hsp) 70 and 90 before heat stress. Meanwhile, after heat stress, up to a fivefold increase was recorded in mRNA level of hsp 70 and fourfold increase in the expression level of hsp 90 in the liver of the control fish which were not fed with MLM-supplemented diets. Fish fed with 1 and 5% MLM showed a significant decrease in the expression of hsp 70 and a non-significant decrease in the expression of hsp 90. Results of the present study suggest that supplementing the diet of grass carp with 5% MLM could improve growth and physiological performance and provide resistance against high temperature-induced stress.

Correlation of insulin resistance with neutrophil to lymphocyte ratio and serum ferritin in male patients of metabolic syndrome.

OBJECTIVE: To determine the correlation of insulin resistance with neutrophil-to-lymphocyte ratio and serum ferritin, and to evaluate whether NLR and serum ferritin can predict insulin resistance in metabolic syndrome. METHODS: The cross-sectional analytical study was conducted at the University of Health Sciences, Lahore, Pakistan, from July 2016 to 2019, and comprised male patients of metabolic syndrome and healthy controls. The correlation involving insulin resistance, serum ferritin and neutrophil-to-lymphocyte ratio was determined. Data was analysed using SPSS 22. RESULTS: Of the 210 subjects, 160(76.2%) were cases with a median age of 45 years (interquartile range: 39-50 years), and 50(23.8%) were controls with a median age of 41 years (interquartile range: 35-50 years). Serum ferritin, alanine aminotransferase, total neutrophil count, lymphocyte count and neutrophil-to-lymphocyte ratio were significantly higher among the cases than the controls (p<0.05). Significant positive correlation of insulin resistance was observed with serum ferritin and neutrophil-to-lymphocyte ratio (p<0.05)) among the cases. Neutrophil-to-lymphocyte ratio significantly predicted insulin resistance among the cases (p<0.05). Conclusion: Neutrophil-to-lymphocyte ratio was fund to be a significant predictor of insulin resistance in metabolic syndrome.

Salivary Irisin and periodontal clinical parameters in patients of chronic periodontitis and healthy individuals: A novel salivary myokine for periodontal disease.

OBJECTIVE: To evaluate changes in the levels of salivary irisin in chronic periodontitis, and to correlate the two. METHODS: The analytical cross-sectional study was conducted at Fatima Memorial Hospital & College of Dentistry, Lahore, Pakistan, from September 2017 to March 2018, and comprised patients of either gender visiting the periodontic out-patient department. The subjects were divided into group I, which had periodontally healthy controls, and group II, which had an equal number of chronic periodontitis patients. Chronic periodontitis was assessed on basis of pocket probing depth, clinical attachment level, plaque percentage and bleeding on probing. Also, 4ml of un-stimulated saliva was collected for the quantification of irisin protein using enzyme-linked immunosorbent assay. Data was analysed using SPSS 25. RESULTS: Of the 40 subjects, there were 20 (50%) in group I with 10 (50%) males and 10 (50%) females having an overall mean age of 37.60±2.58 years. The remaining 20 (50%) subjects were in group II with 16 (80%) males and 4 (20%) females having an overall mean age of 43.25±6.10 years. Mean salivary irisin level in group II was 6.80±3.97ng/ml compared to 3.99±2.48 ng/ml in group I (p=0.009). Periodontal clinical parameters in both the groups were positively but non-significantly correlated with salivary irisin levels (p>0.05) except for decreased plaque percentage in group I (p<0.05). CONCLUSION: Salivary irisin levels increased in chronic periodontitis and decreased with decreasing plaque percentage in healthy individuals, indicating that this myokine can act as a biomarker for chronic periodontal disease.

Pathogenesis of Diabetic Cardiomyopathy and Role of miRNA.

Diabetic cardiomyopathy is characterized as abnormal function and structure of myocardium associated with diabetes irrespective of other cardiac risk factors like hypertension or coronary artery disease (CAD). The pathogenesis of DCM was not well understood in the past due to its complexity but it has been discovered recently. Various factors are found to be associated with the onset of DCM including impaired calcium handling, remodeling of extracellular matrix (ECM), increased oxidative stress, altered metabolism, mitochondrial dysfunction, and endothelial dysfunction. Micro-RNAs (miRNAs) are also found to be of great importance in the pathogenesis of DCM. Different miRNAs like miR-126, miR-24, miR-1, miR-155, miR-499, and miR-199a are found to be associated with different types of heart diseases like CAD and myocardial infarction. Studies have shown that the miRNA plays a crucial role in the development of DCM and it was found that the expression levels of different miRNAs differ in patients as compared to healthy individuals. This review focuses on the pathogenesis of DCM and various factors involved in the onset of diabetic car-diomyopathy. Moreover, the probable role of miRNA in the pathogenesis of DCM is also discussed.

Serum resistin levels as predictor of low bone mineral density in postmenopausal women.

Resistin, a novel adipokine may play an important role in bone metabolism. The study is designed to discover the association of bone mineral density (BMD) with serum resistin levels, anthropometric measures and to elucidate serum resistin as a predictor of BMD in postmenopausal women. Postmenopausal women (n = 160) were recruited and divided into two groups, non-osteoporotic (n = 70) and osteoporotic (n = 90). BMD was evaluated by DXA scan. High serum resistin levels and low weight are independent contributors to low BMD and can influence BMD at lumbar spine, right femoral neck, right hip, left femoral neck, and left hip in postmenopausal women.

Association between hypoxia-inducible factor-1 alpha rs11549465 (1772 C>T) polymorphism and metabolic syndrome.

OBJECTIVE: To find the association of single nucleotide polymorphism of hypoxia-inducible factor-1 alpha, rs11549465 (1772 Cytosine > Thymine) with metabolic syndrome, and to compare the anthropometric and biochemical variables in different genotypes of hypoxia-inducible factor-1 alpha. METHODS: The cross-sectional comparative study was conducted at the University of Health Sciences, Lahore, Pakistan, from July 2016 to April 2019, and comprised patients of metabolic syndrome selected from the Sheikh Zayed Hospital, Lahore. Healthy controls were also enrolled. Fasting venous sample was taken for the determination of study parameters. The genetic variant of hypoxia-inducible factor-1 alpha was analysed by restriction fragment length polymorphism polymerase chain reaction. Data was analysed using SPSS 22. RESULTS: Out of 400 subjects, 200(50%) each were patients and controls. The frequency of CC genotype of hypoxia-inducible factor-1 alpha Cytosine > Thymine in patients was 166(83%) and in controls 147(73.5%); CT genotype was 34(17%) and 53(26.5%) respectively, while TT genotype was not observed. There was a significant association of the C allele and CC genotype (p=0.03) with the increased risk of metabolic syndrome (p=0.02). On comparison of study variables in the two genotypes, systolic blood pressure, anthropometric and lipid parameters were significantly higher in the wild CC genotype compared to CT in the control group (p<0.05), but there was no significant difference in the patients (p>0.05). CONCLUSIONS: Major allele C of hypoxia-inducible factor-1 alpha 1772 Cytosine > Thymine was found to be associated with increased risk of metabolic syndrome.

Expression levels of MiRNA-16, SURVIVIN and TP53 in Preeclamptic and Normotensive women.

OBJECTIVE: To measure and compare micro ribonucleic acid-16, survivin and tumour protein p53-regulated apoptosis-inducing protein 1 expression levels in preeclamptic and normotensive pregnancies, and to check the correlation of micro ribonucleic acid-16 with messenger ribonucleic acid expression of survivin and tumour protein p53. METHODS: The observational cross-sectional comparative study was conducted at the Department of Physiology and Cell Biology, University of Health Sciences, Lahore, Pakistan, from 2016 to 2018, and comprised preeclamptic women in group A and normotensive women in group B. The preeclamptic patients were further categorised into early-onset preeclampsia subgroup A1and late-onset preeclampsia group A2. Expression of micro ribonucleic acid-16, messenger ribonucleic acid expression of survivin and tumour protein p53 in preeclamptic and normotensive pregnancies were analysed using real time polymerase chain reaction. Data was analysed using SPSS 22. RESULTS: Of the 54 patients, 27(50%) were in each of the two groups. Within group A, 14(52%) patients were in group A1 and 13(48%) in group A2. The expression of micro ribonucleic acid 16 showed significant increase in group A compared to group B (p<0.05). The difference was not significant between the subgroups A1 and A2. The levels of messenger ribonucleic acid expression of survivin and tumour protein p53 were deregulated in group A, with a decrease in survivin and an increase in tumour protein p53. The messenger ribonucleic acid expression of survivin and tumour protein p53 showed statistically significant differences across subgroups A1 and A2 (p<0.05). The micro ribonucleic acid-16 expression correlated negatively with messenger ribonucleic acid expression of survivin, but exhibited a positive correlation with tumour protein p53. CONCLUSIONS: Deregulated micro ribonucleic acid-16 along with differentially expressed apoptotic genes, survivin and tumour protein p53 might result in altered apoptosis implicated in the pathogenesis of preeclampsia.

TCF7-L2 rs7903146 polymorphism in metabolic syndrome with and without acute coronary syndrome.

OBJECTIVE: To determine the frequency and association of single nucleotide polymorphism of transcription cell factor7-like2 rs7903146 (C>T) in metabolic syndrome patients with and without acute coronary syndrome. METHODS: The cross-sectional comparative study was conducted at the University of Health Sciences, Lahore, Pakistan, from July to December 2017. Patients of metabolic syndrome with and without acute coronary syndrome were selected from Sheikh Zayed Hospital, Lahore, and the Punjab Institute of Cardiology, Lahore. Healthy subjects were enrolled to act as controls. A fasting blood sample of 8ml was taken for deoxyribonucleic acid extraction and estimation of biochemical parameters. Single nucleotide polymorphism of transcription cell factor7-like2 rs7903146 C>T was determined using restriction fragment length polymorphism. SPSS 22 was used for data analysis. RESULTS: Of the 500 subjects, 200(40%) were group A patients without acute coronary syndrome, 100(20%) were in group B with acute coronary syndrome and 200(40%) were group C controls. Overall, 385(77%) were males and 115(23%) were females. The frequency of CC variant in group A was 35(17.5%) and in group C 22(11%), while CT was 32(16%) and 65(32.5%), and TT was 133(66.5%) and 113(56.5%), respectively. There was significant association of TT genotype with increased risk of metabolic syndrome (p=0.031), and CC genotype had no association (p=0.121). There was no significant difference of genotype frequency between groups A and B (p=0.246), but TT variant was significantly higher in group A compared to group B (p=0.009). CONCLUSIONS: TT genotype of transcription cell factor7-like2 rs7903146 C>T was found to be associated with increased risk of metabolic syndrome in patients without acute coronary syndrome compared to those with acute coronary syndrome and healthy controls.

Validation of Housekeeping Genes for Gene Expression Profiling in Fish: A Necessity.

Real-time polymerase chain reaction (qPCR) is a reliable tool for gene expression analysis in various organisms. Its use in aquaculture and study of fish physiology is increasing day by day. Relative quantification of mRNA through real-time qPCR requires suitable housekeeping genes/reference genes. Expression of housekeeping genes were thought to be constant in different tissues under various physiological and experimental conditions. In light of the literature reviewed here, it is clear that expression of most of the genes varied in different physiological conditions, under different experimental conditions, and in different gender. Use of housekeeping genes without proper validation may lead to erroneous results and misinterpretation of data. To solve this problem, various types of user-friendly software are now available for validation of housekeeping genes. Moreover, it is recommended to use more than one housekeeping gene for relative quantification. Using the geometric mean of the three most stable genes as a housekeeping gene can also solve this problem to some extent. Therefore, there is a dire need to validate a set of housekeeping genes in the species under study and in various tissues depending on the experimental design to obtain valid and reproducible results.

Adrenergic receptor beta-3 rs4994 (T>C) and liver X receptor alpha rs12221497 (G>A) polymorphism in Pakistanis with metabolic syndrome.

The present study aimed to determine the association of adrenergic receptor beta-3 (ADRB3) rs4994 T>C and liver X receptor alpha (LXR-α) rs12221497 G>A polymorphism with metabolic syndrome (Met S) and the related traits in Pakistanis. Patients of Met S were recruited from the Endocrinology and Diabetic Clinic of Sheikh Zayed Hospital Lahore, over the time span of 6 months from July to December 2016. Single-nucleotide polymorphism (SNP) of ADRB3 was determined by restriction fragment length polymorphism and of LXR-α by amplification refractory mutation system polymerase chain reaction. The frequency of TT variant of ADRB3 T>C in Met S was 69 (34.5%) and in controls 89 (44.5%), frequency of TC 103 (51.5%) and 96 (48%), and of CC 28 (14%) and 15 (7.5%), respectively. In the recessive model (CC: TT + TC), CC genotype was found to be associated with the increased risk of Met S (P = 0.027; odds ratio [OR] = 2.09; confidence interval [CI] =1.08-4.03) and the association remained significant after controlling for the confounders such as age and sex. The frequency of GG variant of LXR-α G>A in Met S was 35 (17.5%) and in controls 15 (7.5%), GA 129 (64.5%) and 137 (68.5%), and AA 36 (18%) and 48 (24%), respectively. In the recessive model (GG: GA + AA), GG genotype was found to be associated with the increased risk of Met S (P = 0.004; OR = 2.52; CI = 1.33-4.80) and the association remained significant after controlling for the confounders such as age and sex. It was concluded that SNP of ADRB3 (190 T>C) and LXR-α (-115 G>A) were associated with the risk of Met S and might increase the susceptibility to the obesity-related traits.

Altered expression of vascular endothelial growth factor, vascular endothelial growth factor receptor-1, vascular endothelial growth factor receptor-2, and Soluble Fms-like Tyrosine Kinase-1 in peripheral blood mononuclear cells from normal and preeclamptic pregnancies.

Preeclampsia (PE) is the leading cause of maternal and fetal morbidity and mortality. It complicates around 2%-10% pregnancies worldwide due to imbalance between proangiogenic and anti-angiogenic factors, leading to incomplete placentation, ischemia, and endothelial dysfunction. The study was aimed to analyze the mRNA expression of vascular endothelial growth factor (VEGF) and its receptors, i.e., VEGF receptor-1 (VEGFR-1), VEGF receptor-2 (VEGFR-2), and soluble Fms-like tyrosine kinase-1 (sFlt-1) from maternal peripheral blood mononuclear cells (PBMCs) of PE patients. This was a cross-sectional comparative study comprising 18 normotensive and 18 PE patients; the patients were further divided as early-onset preeclampsia (EOP) and late-onset preeclampsia (LOP). The expression level of VEGF, its receptors (VEGFR-1 and VEGFR-2), and sFlt-1 was investigated using real-time polymerase chain reaction. There was a significant change in the mRNA expression with a decrease in VEGF, VEGFR-1, and VEGFR-2 and an increase in sFlt-1 in PBMCs of PE and normal pregnancies (P < 0.001). sFlt-1 mRNA expression was increased by 2.95-fold in the PE group with an inverse correlation with expression of VEGFR-2 (Spearman's rho = 0.68). Based on these findings, we conclude that PE is associated with decrease in the mRNA expression of VEGF, VEGFR-1, and VEGFR-2 as compared to an increase in sFlt-1 in PBMCs.

Modulation of brain kisspeptin expression after bisphenol-A exposure in a teleost fish, Catla catla.

Endocrine-disrupting chemicals (EDCs) affect the neuroendocrine system which in turn influences the reproductive regulation. Neuronal genes disrupted by EDCs are the gonadotropin-releasing hormone (gnrh2), the Kiss/GPR54 system that regulates gonadotropin release and cyp19b gene encoding brain aromatase. In the present study, pubertal Catla catla expected to spawn for first the time in the coming season were exposed to graded concentration of bisphenol-A (10, 100, 1000 µg/l) for 14 days. Messenger RNA (mRNA) levels of neuroendocrine genes, i.e., kisspeptins and their receptors, gonadotropin-releasing hormone type II and brain aromatase were studied after 14 days exposure. Results showed that bisphenol-A (BPA) strongly upregulated expression of kiss1, kiss2, gpr54a, and gnrh2 in fish exposed to 10 µg/l BPA. Fish exposed to 1000 µg/l BPA, expression of kiss1 and gnrh2 were comparable to control while kiss2 mRNA increased compared to controls. Brain aromatase (cyp19b) mRNA expression increased in fish exposed to both 10 and 1000 µg/l BPA. These results indicate that BPA exposure can disrupt organization of the kisspeptin signaling pathways. This neuroendocrine disruption may be the underlying mechanism by which a suite of reproductive abnormalities are induced.

Peroxisome proliferator activated receptor gamma 34C>G variant and anthropometric parameters in metabolic syndrome.

OBJECTIVE: To determine the frequency of 34 Cytosine >Guanine (proline 12 alanine) variant of peroxisome proliferator activated receptor gamma, and to associate it with metabolic syndrome, insulin resistance and anthropometric obesity parameters. METHODS: The cross-sectional comparative study was conducted at the University of Health Sciences, Lahore, Pakistan, from September 2016 to 2017, and comprised patients of metabolic syndrome and healthy controls. Blood pressure and anthropometric measurements of all the subjects were recorded. Fasting blood sample of 4ml was taken for biochemical parameter and deoxyribonucleic acid extraction. The frequency of genetic variant was determined by amplification refractory mutation system polymerase chain reaction. Data was analysed using SPSS 22. RESULTS: Out of 400 subjects, 200 (50%) each were patients and controls. Overall, there were 308 (77%) males and 92 (23%) females. Patients had significantly higher blood pressure, body mass index, waist circumference, waist-to-hip ratio, mid-arm circumference and triceps skinfold thickness compared to the controls (p<0.0001). Insulin resistance was also significantly higher in the patients (p<0.0001) and showed significant correlation with body mass index, waist circumference, waist-to-hip ratio, mid-arm circumference and triceps skinfold thickness (p<0.05).Waist circumference and triceps skinfold thickness were significant predictors of homeostatic model assessment for insulin resistance. Overall, the frequency of homozygous dominant genotype CC of PPAR2 34C>G was 291 (72.75%), heterozygous CG was 93 (23.25%) and homozygous recessive GG was 16 (4%).There was no significant difference in frequency of genotypes between the groups (p=0.216). However, waist circumference and body mass index were significantly lower in GG genotype compared to the CC (p=0.006 versus p=0.02). CONCLUSIONS: Waist circumference and triceps skinfold thickness were found to be the significant predictors of homeostatic model assessment for insulin resistance, while no association was found between 34 C>G variant of peroxisome proliferator activated receptor gamma and metabolic syndrome.

Alkaline phosphatase is a predictor of Bone Mineral Density in postmenopausal females.

OBJECTIVES: The study was planned to determine whether serum calcium, phosphate and alkaline phosphatase (ALP) are predictors of bone mineral density (BMD) in postmenopausal non-osteoporotic, osteopenic, and osteoporotic females. METHODS: In this cross sectional study, conducted at Shaikh Zayed Hospital, Lahore in the year 2014-2015, postmenopausal females between 50-70 years of age were taken and divided into three groups non-osteoporotic (n=52), osteopenic (n=69) and osteoporotic (n=47). Serum ALP, phosphate and calcium were used in a stepwise multiple regression analysis to predict T-score in these groups. RESULTS: In normal postmenopausal females, the prediction model was statistically significant, F(2, 41) = 6.041, p < 0.05 and showed a T-score variance of 22%. T-score was primarily predicted by higher levels of phosphate and calcium. In postmenopausal osteopenic females, T-score was only predicted by lower levels of ALP. The model was statistically significant, F(1, 59) = 4.995, p < 0.05, and accounted for approximately 7% of the variance of T-score. In postmenopausal osteoporotic females, the prediction model contained no predictors. CONCLUSION: Our study suggested that calcium and phosphate are the strongest predictors of T-score in postmenopausal normal females, while in postmenopausal osteopenic females ALP was the strongest predictor of T-score. Elevated serum ALP levels may help in determining loss of BMD in postmenopausal females.

Distribution of IL28B and IL10 polymorphisms as genetic predictors of treatment response in Pakistani HCV genotype 3 patients.

There are over 10 million hepatitis C virus (HCV)-infected patients in Pakistan. For these patients, a combination of interferon with ribavirin is the most economical and easily available treatment. Single-nucleotide polymorphisms in interleukin genes have been reported to be associated with the pathogenesis and clearance of HCV, and sustained virologic response (SVR). An interleukin 28B (IL28B) gene polymorphism has been shown to modify treatment outcomes, but the effects of interleukin 10 (IL10) polymorphisms have not been previously assessed in the Pakistani population. The present study was conducted with 302 subjects categorized into two groups: 100 healthy volunteers (Group I) and 202 patients with chronic HCV (Group II). Patients within Group II were further divided into two subgroups according to therapeutic response: SVR (responders = 132) and NR (non-responders/relapsers = 70). IL28B (rs8099917, rs12979860) and IL10 (rs1800872, rs1800871, rs1800896) gene polymorphisms were studied in all subjects. A significant difference in the distribution of IL28B rs12979860C/T genotypes between the two groups (p<0.05) was observed, while of the three IL10 polymorphisms, a significant difference was only shown for rs1800896 A/G. Haplotype analysis (IL28B and IL10) showed a significant association of TTGTC and TTGTA when comparing the groups. There was a strong association of the favorable alleles rs8099917T and rs12979860C in the SVR group as compared with the NR group (p<0.05), and rs1800896 also showed an association with the SVR group as compared to the NR group (p<0.004). Haplotype analysis showed significant associations when comparing the SVR and NR subgroups, i.e. TCATC (p=0.009), TTGTA (p=0.005), TCATA (p<0.0005), TCACA (p=0.002), GTGCC (p=0.002) and TCGTC (p=0.005). IL28B (rs8099917 and rs12979860) and IL10 (rs1800896) polymorphisms alone, or in combination, are good predictors of therapeutic response in HCV-3a patients.

Association of TP53 codon 72 polymorphism in women suffering from endometriosis from Lahore, Pakistan.

OBJECTIVE: To investigate TP53 gene codon 72 polymorphism in women with endometriosis and compare it with healthy samples. METHODS: This case-control study was carried out at Jinnah Hospital, Services Hospital and Sheikh Zayed Hospital, Lahore, Pakistan, from 2014 to 2016, and comprised patients with endometriosis and healthy controls. SPSS 21 was used for statistical analysis. RESULTS: Of the 176 participants, 88(50%) were healthy controls and 88(50%) were endometriosis patients. The observed genotype frequencies for controls and patients were 14(15.9%) and 31(35.3%) for proline/proline, 46(52.3%) and 35(39.8%) for proline/arginine, and 28(31.8%) and 22(25%) for arginine/arginine, respectively. The association of different genotypes was not significant in patients with moderate-to-severe endometriosis (p=0.574). The presence of pro/pro genotype enhanced the chances/odds of getting the disease (p<0.05). However, the risk further increased with the advancement of age, particularly in the 27-46 age group (p<0.05). CONCLUSIONS: In Pakistani women the association of TP53 gene codon 72 arginine/proline polymorphism was present..

Current Status of Direct Acting Antiviral Agents against Hepatitis C Virus Infection in Pakistan.

In Pakistan, the burden of the hepatitis C virus (HCV) infection is the second highest in the world with the development of chronic hepatitis. Interferon-based combination therapy with ribavirin was the only available treatment until a few years back, with severe side-effects and high failure rates against different genotypes of HCV. Interferon-free all-oral direct-acting antiviral agents (DAAs) approved by the FDA have revolutionized the HCV therapeutic landscape due to their efficiency in targeting different genotypes in different categories of patients, including treatment naïve, treatment failure and relapsing patients, as well as patients with compensated and decompensated cirrhosis. The availability and use of these DAAs is limited in the developing world. Sofosbuvir (SOF), a uridine nucleotide analogue and inhibitor of HCV encoded NS5B polymerase, is now a widely available and in-use DAA in Pakistan; whereas daclatasvir was recently added in the list. According to the documented results, there is hope that this disease can be effectively cured in Pakistan, although a few concerns still remain. The aim of this article is to review the effectiveness of DAAs and the current status of this treatment against HCV genotype 3 infection in Pakistan; various factors associated with SVR; its limitations as an effective treatment regime; and future implications.

Analysis of toll-like receptors-9 (TLR9) gene polymorphism (rs5743836) in Pakistani patients with HCV.

Toll-like receptors (TLRs) are innate immune receptors that mediate the inflammatory response during HCV infections. The goal of this study was to evaluate the association of TLR9 gene polymorphism (rs5743836) in Pakistani patients infected with genotype 3a of HCV. Total 500 subjects were recruited, 400 HCV patients and 100 healthy individuals. Genotyping of TLR9 (-1237T/C, rs5743836) was carried out in 400 HCV patients (323 interferon responders and 77 interferon non responder) and control group by applying High resolution melting (HRM) curve assay. No remarkable differences in distribution of genotype between HCV (p<0.0001; OR= 3.21, 95% CI= (2.514.12) and control groups (p<0.0001; OR=0.092, 95%CI= (0.0580.14) were observed. In conclusion TLR9-1237T/C gene polymorphism may not be considered as a molecular risk for patients with HCV in Pakistan.