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Background The severity and complexity of autoimmune disorders is dependent on the genetic capability of individuals. Genetic studies have revealed association between polymorphisms of Vitamin D Receptor gene and individuals’ predisposition to autoimmune diseases. Therefore, this study aimed to develop relationship between vitamin D receptor gene polymorphisms and hypothyroidism. Materials and Methods A total of 144 individuals were studies, including 72 patients presenting with symptoms of Hashimoto’s disease. The amplicon sequencing was performed on samples bearing M13 tail tags. Statistical analysis was carried out using SPSS software to establish correlations of genotypes and alleles among control and diseased individuals. Results The mean concentrations of vitamin D were observed to be critically low in patients with Hashimoto’s thyroiditis. Of the four SNPs studied, only rs7975232 was found to be significantly related with disease progression. Other three rs1544410, rs731236 and rs2228570 did not show significant correlation in the individuals studied. Conclusion Altered VDR expressions because of various VDR polymorphisms have been shown to exhibit differently among various races and ethnic groups. Furthermore, these VDR polymorphisms also vary among populations in different environmental and genetic predispositions. This study suggests the homozygous CC genotype at rs7975232 to be more as a risk factor for development of Hashimoto’s disease in the population studied.

Objective: To assess the association of miR-146a and its target protein RhoA expression levels in breast cancer. Methods: The case-control study was conducted at Riphah International University, Islamabad, Pakistan, from March 2017 to November 2018, and comprised confirmed breast cancer cases and controls who were matched for age and ethnicity. Genotyping and expression profiling of archived samples was performed. Data was analysed using SPSS 22.. Results: Of the 590 subjects, 295(50%) each were cases and controls. Among the cases, there were 195(66%) Punjabis, 59(20%) Pathans and 41(14%) Kashmiris. The corresponding numbers among the controls were 198(67%), 58(19.7%) and 39(13.2%). The association between genotypes of the cases and controls was significant (p0.05). No association was found with the receptor status and miR-146a polymorphism. Conclusion: The miR-146a gene polymorphism rs2910164 G/C was found to have increased susceptibility to breast cancer at genotype and allelic levels. Key Words: Single nucleotide polymorphism, Breast cancer, miR-146a, RhoA.

OBJECTIVE To investigate the association of receptor for advanced glycation end products gene polymorphism 429T/C (rs1800625) with diabetic retinopathy and serum soluble receptor for advanced glycation end products levels in patients with type 2 diabetes. METHODS The case-control study was conducted from January 2017 to December 2018 at Pakistan Railway Hospital, Rawalpindi, and the Multidisciplinary Laboratories of Islamic International Medical College, Riphah International University (RIU), Islamabad, Pakistan. Those included were healthy controls in group A, diabetics without retinopathy in group B and patients having diabetic retinopathy in group C. Genotyping for 429T/C was done by tetra-primer amplification refractory mutation system-polymerase chain reaction. Serum soluble receptor for advanced glycation end products levels were measured using enzyme-linked immunosorbent assay. Data was analysed using SPSS 22. RESULTS Of the 450 subjects, 150(33.3%) were in each of the three groups. The frequency of TT, TC and CC genotypes of 429T/C polymorphism were 137(91.3%), 10(6.7%) and 3(2%) in group A; 133(88.6%), 13(8.7%) and 4(2.7%) in group B; and 127(84.7%), 18(12%) and 5(3.3%) in group C. No significant association of 429T/C genotypic and allelic frequencies were found with groups B and C (p>0.05). Serum soluble receptor for advanced glycation end products levels were significantly high in patients with proliferative diabetic retinopathy and were positively correlated with fasting plasma glucose in group C (p

Objectives: To determine differential expression of microRNAs (miRNAs) in plasma of 2, 4, 6-Trinitrotoluene (TNT) exposed ordnance factory workers. Methods: A case control study was conducted at the Department of Toxicology, Armed Forces Institute of Pathology, Rawalpindi from July to December 2020. A total 30 subjects were recruited from an ordnance factory that were directly exposed to TNT and 120 non-exposed individuals from non-factory healthy population. Plasma levels of five miRNAs including miRNA-let-7a-2, miRNA-34a-1, miRNA-21-2, miRNA-106b-1, miRNA-122a-1 were measured by quantitative real-time polymerase chain reaction (RT-PCR). Results: Micro RNAs showed a wide range of Ct (cycle threshold) values ranging from 23.48 to 41.94. Among the five miRNAs let-7a-2 and miRNA-122a-1 displayed relatively high expression with Ct values ranging from 26.58 ± 2.25 to 27.18 ± 0.80 respectively. Relative fold change expression for all five miRNAs of exposed individuals were found high (p 10, the individuals having ≤ 10-fold change expression were19 (63.3%) in miRNA-let-7a-2, 30 (100%) in miRNA-34a-1 and 23 (76.7%) in miRNA-122a-1 while in miRNA-21-2 and miR-106b-1, 23 (76.7%) and 18(60%) individuals had > 10-fold change expression respectively. Among the five miRNAs in exposed individuals, miRNA-let-7a-2, miR-21-2, miR-106b-1 and miR-122a-1 were found highly expressed with fold change expression > 10 (p

To investigate the association of receptor for advanced glycation end products gene polymorphism 429T/C (rs1800625) with diabetic retinopathy and serum soluble receptor for advanced glycation end products levels in patients with type 2 diabetes.The case-control study was conducted from January 2017 to December 2018 at Pakistan Railway Hospital, Rawalpindi, and the Multidisciplinary Laboratories of Islamic International Medical College, Riphah International University (RIU), Islamabad, Pakistan. Those included were healthy controls in group A, diabetics without retinopathy in group B and patients having diabetic retinopathy in group C. Genotyping for 429T/C was done by tetra-primer amplification refractory mutation system-polymerase chain reaction. Serum soluble receptor for advanced glycation end products levels were measured using enzyme-linked immunosorbent assay. Data was analysed using SPSS 22.Of the 450 subjects, 150(33.3%) were in each of the three groups. The frequency of TT, TC and CC genotypes of 429T/C polymorphism were 137(91.3%), 10(6.7%) and 3(2%) in group A; 133(88.6%), 13(8.7%) and 4(2.7%) in group B; and 127(84.7%), 18(12%) and 5(3.3%) in group C. No significant association of 429T/C genotypic and allelic frequencies were found with groups B and C (p0.05). Serum soluble receptor for advanced glycation end products levels were significantly high in patients with proliferative diabetic retinopathy and were positively correlated with fasting plasma glucose in group C (p0.05). TC and CC genotypes were significantly associated with raised serum soluble receptor for advanced glycation end products, and TC with raised fasting plasma glucose in group C.The 429T/C receptor for advanced glycation end products gene polymorphism was found to be associated with severe non-proliferative diabetic retinopathy, and serum soluble receptor for advanced glycation end products levels had a positive correlation with severity of diabetic retinopathy.

Objectives: to determine the relationship of 374T/A (rs1800624) polymorphism in the gene encoding RAGE with Type-2 diabetes mellitus (T2DM), diabetic retinopathy (DR) and serum soluble RAGE (sRAGE) level in Pakistani patients. Methods: A case-control study, conducted from January 2017 to December 2018, involving 150 healthy controls (HC), 150 T2DM patients with no retinopathy (DNR) and 150 DR patients diagnosed by coloured fundus photography. Tetra-primer amplification refractory mutation system – polymerase chain reaction (T-ARMS-PCR) was used for genotyping. Serum sRAGE levels were measured by enzyme-linked immunosorbent assays (ELIZA). Results: The frequency of TT, TA and AA genotypes of rs1800624 polymorphism were: 92.7%, 6%, 1.3% in HC, 80%, 17.3%, 2.7% in DNR and 76.7%, 19.3%, 4.3% in DR groups. Heterozygous TA genotype and mutant A allele showed significant association with diabetes and DR vs HC. In dominant model, mutant allele showed significant association with DNR and DR vs HC. No significant association of rs1800624 was detected with DR and its sub-groups, non-proliferative DR (NPDR) and proliferative DR (PDR) vs DNR. Dividing NPDR into mild, moderate and severe, heterozygous TA genotype showed significant association with moderate and severe NPDR vs DNR. In DNR and DR groups, TA genotype was significantly associated with raised sRAGE. Conclusion: rs1800624 RAGE gene polymorphism might be a risk factor for T2DM and NPDR in Pakistani patients. Raised sRAGE levels have a positive correlation with PDR and are associated with heterozygosity of rs1800624 polymorphism in DNR and DR groups. doi: https://doi.org/10.12669/pjms.37.3.3670 How to cite this:Qayyum S, Afzal M, Naveed AK. Association analysis of 374T/A (rs1800624) receptor for advanced glycation end-products (RAGE) gene polymorphism with diabetic retinopathy in Pakistani patients. Pak J Med Sci. 2021;37(3):733-739. doi: https://doi.org/10.12669/pjms.37.3.3670 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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OBJECTIVE To determine the association of single nucleotide polymorphism in three CC, TT and TC genotypes of transforming growth factor β1 T29C in breast cancer patients. METHODS The case-control study was conducted from April 2017 to April 2018 at the Islamic International Medical College, Rawalpindi, Pakistan, in collaboration with Nuclear Oncology Medicine and Radiotherapy Institute and Holy Family Hospital, Rawalpindi. Using convenience sampling, breast cancer cases and healthy controls were enrolled. All investigations were done using standardized laboratory protocols. The outcomes were determined in terms of association of single nucleotide polymorphism of transforming growth factor β1with breast cancer. Data was analysed using SPSS 21. RESULTS Of the 150 subjects, 80(53.3%) were cases and 70(47.7%) were healthy controls. Among the cases, the most frequent genotype was CC 38(47.5%) followed by TC 26(32.5%) and TT 16(20%). Among the controls, the corrsesponding values were 50(71.42%), 13(18.5%) and 7(10%). Transforming growth factor β1 TC genotype was strongly associated with the increased risk of developing breast cancer (odds ratio: 3.79). CONCLUSIONS The incidence of breast cancer was markedly lower among women with CC genotype compared to those with CT or TT genotypes.

Background: CK2, a serine/threonine, protein kinase, targets over and above300 substrates including c-Myc. CK2 expression is elevated in human cancers includingbreast cancer and prostate cancer. c-Myc protooncogene expression is also up-regulated inthese cancers. Objectives: To evaluate the co expression and correlation of CK2 and c-Mycin prostate cancer as compared to their correlation in breast cancer. Study Design: Crosssectional analytical study. Setting: Army Medical College and AFIP, Duration: Two years.Methods: A retrospective study of immunohistochemical analysis, approved by Armed ForcesInstitute of Pathology Ethical Committee. Paraffin embedded tissues of diagnosed prostatecancer, 30 in number, 30 cases of Benign Prostatic Hypertrophy (BPH) and 30 cases of breastadenocarcinoma, were included in the study. We stained tissue sections for CK2 and c-Mycand measured staining intensity for each protein expression. Data analysis was done by SPSSversion 20. Pearson correlation coefficient was used for correlating the expression of bothproteins. P-value was calculated. Results: A strong correlation of CK2 with c-Myc was seen inprostate cancer tissue, in comparison to BPH. There was a very significant correlation presentbetween CK2 and c-Myc, especially in invasive cases of breast cancer. Conclusion: CK2 andc-Myc expressions are highly and significantly correlated in prostate cancer and breast cancerespecially in invasive cases. CK2 has influence over c-Myc and both can be used for forecastingthe cancer phenotype and aggression of disease.

Introduction: Integrative genomics may help in the identification of novelbiological pathways in the pathogenesis of CAD. Objectives: To find out the association of 5Cytokine SNPs and 13 CAD SNPs gene risk scores with serum cytokine levels in PrematureCoronary Artery Disease (PCAD). To identify the direct and indirect protein interactions of the 13CAD risk genes and the 5 cytokine genes in PCAD. Study Design: Case-control study. Setting:Army Medical College, in association with University College London (UCL), London, UnitedKingdom (UK). Materials and Methods: 340 PCAD patients and 310 age and sex matchedcontrols were recruited. Serum IL18, TNFA, IL6 and IL10 levels were measured using ELISA(Invitrogen). The SNPs were genotyped using TAQMAN and KASPar assays. Data analysis wasdone using standard SPSS software version-21 (SPSS Inc, Chicago, Illinois, USA). The proteinproteininteraction (PPI) network was generated using STRING version 9.0, Genemania andI-Tessar web. Results: The patients of PCAD had mean ± SD age of 42 ± 3.80 years consistingof 329 males and 11 females. The 5 SNP cytokine gene risk score correlated significantly withthe serum IL-18, IL-6, TNF-alpha, IL-18: IL-10 and TNF-alpha: IL-10 ratios (p

Objectives: To study the effect of maternal high-fat, high-cholesterol diet on thelipid profiles of rat offspring. Study Design: Randomized control trial (RCT). Place and durationof study: Department of Biochemistry, Army Medical College, Rawalpindi in collaborationwith Chemical Pathology Laboratory (CPL), Army Medical College, Rawalpindi and NationalInstitute of Health (NIH), Islamabad. The study was completed in six months. Material andMethods: Sixty adult female albino rats of Sprague-Dawley strain were divided into two groupsof 30 each. Male rats of the same strain were used for breeding. Ten days before expectedmating experimental group was shifted to a high-fat, high-cholesterol diet, keeping the controlgroup on normal rat diet. Lipid profiles of pregnant dams of both groups were done at day20 of gestation. Offspring of both groups from weaning onwards were fed the normal rat diet.One male and one female offspring from each litter of both groups (30 male and 30 femalesfrom each group) were selected and their lipid profiles were evaluated at post-weaning andadult stage. Results: At day 20 of gestation, high-fat and cholesterol fed dams showed highlysignificant increase in TC, LDL-c, HDL-c and LDL-c / HDL-c ratio than the control dams (p

Vanadyl sulphate is available as herbal medicine against diabetes mellitus and body building supplement, over the counter worldwide. The available data on its safety is controversial and inadequate. The objective of this study was to analyse its safety in usual therapeutic dose range.It was an experimental study carried out at the Department of BiochemistryMolecular Biology, Army Medical College, National University of Medical Sciences (NUMS), Rawalpindi, Pakistan, from Jun 2014 to Oct 2018. The study was carried out on 105 Sprague Dawley rats for duration of 24 weeks. The animals were randomly distributed in three groups of 35 each. The group I rats were marked as control while rats of group IIIII were administered vanadyl sulphate 0.06mg/day and 0.3mg/day respectively. Alanine amino transferase (ALT) and Malondialdehyde (MDA) were measured in serum while comet assay was performed on WBCs.The plasma levels of ALT and MDA were significantly raised in group II and III subjects. Single cell gel electrophoresis (SCGE) / comet assay showed minimal "tail moment" in control group and increased tail moment in group II and III in a dose dependent manner which indicates dsDNA breaks.It was observed that vanadyl sulphate causes hepatocellular toxicity, oxidative stress and damage to the DNA in usual therapeutic/ supplemental doses. Due to hazardous effects, its use in humans as alternate medicine may be reviewed.

Objective: To evaluate the association of miR-196a rs11614913 C/T genetic variation and its target gene annexin A1 mRNA expression with breast cancer risk in Pakistani female ethnicities. Methods: This case control study, conducted from March 2017 to November 2018 included 295 breast cancer patients, 295 controls of three Pakistani ethnicities and archived 100 samples of cohort group for genotyping and expression profiling. Genotyping of miR-196a (rs11614913 C/T) was done by ARMS PCR technique. Annexin-A1 (ANXA1) mRNA expression was measured with qRT-PCR and detection of protein expression of ANXA1 was done by immunohistochemistry. Results: CC homozygous genotype of miR-196a rs11614913 was present in 81.4% of cases and 73.9% controls. C/T polymorphism was found to be significantly associated with decrease risk of breast cancer (OR=0.25 (0.11- 0.58, p

Objective: To assess the knowledge and practice of gynecologists in relation to dental and oral healthcare during pregnancy. Material and methods: This cross sectional survey was conducted among gynecologists of Lahore and Bahawalpur from 3-9-19 to 9-9-19. Total 200 gynecologists (from private and government hospitals) were selected for this study. A self-structured questionnaire was distributed to all the selected gynecologists and then collected 3–4 days after the first visit. Knowledge and practice of gynecologists in relation to dental and oral healthcare during pregnancy was assessed. Results: In present study, mean age of the gynecologists was 47.25 years. Total 126 (63%) gynecologists were between 41-50 years. Total 86 (43%) having experience >10 years. About 64% gynecologists agreed that pregnancy increases the likelihood of gingival inflammation. 72% reported that their patients come with bleeding gums, small swelling, and tooth mobility during pregnancy. About 27% gynecologists believe that local anesthetic solution containing vasoconstrictor for pregnant patients is safe. Gingival periodontal inflammation can affect the outcome of pregnancy believed by 63% respondents. Conclusion: Results of present study revealed that majority of gynecologists were having good knowledge, attitude and practices but still there is a need for active participation and more involvement of medical specialists like gynecologists and pediatricians, in continuing education programs and forums on dentistry. Keywords: Oral health, low birth weight, pregnancy, gynecologist.

Objective: To evaluate the association of miR-196a rs11614913 C/T genetic variation and its target gene annexin A1 mRNA expression with breast cancer risk in Pakistani female ethnicities. Methods: This case control study, conducted from March 2017 to November 2018 included 295 breast cancer patients, 295 controls of three Pakistani ethnicities and archived 100 samples of cohort group for genotyping and expression profiling. Genotyping of miR-196a (rs11614913 C/T) was done by ARMS PCR technique. Annexin-A1 (ANXA1) mRNA expression was measured with qRT-PCR and detection of protein expression of ANXA1 was done by immunohistochemistry. Results: CC homozygous genotype of miR-196a rs11614913 was present in 81.4% of cases and 73.9% controls. C/T polymorphism was found to be significantly associated with decrease risk of breast cancer (OR=0.25 (0.11- 0.58, p

Polymorphisms in vitamin K epoxide reductase complex subunit 1 (VKORC1) gene lead to interindividual variability in warfarin dose requirement. The characterization of genotype frequency distribution is required in different populations for construction of customized dosing algorithms to enhance the efficacy and reduce the toxicity of warfarin therapy. This study was carried out in Pakistani population to evaluate the contribution of common VKORC1 polymorphisms to warfarin therapy. A total of 550 stable patients taking warfarin were enrolled after medical history, physical examination, and laboratory investigations. Single blood sample was collected after informed consent. Genomic DNA was extracted and genotype analysis for VKORC1 1173C>T and VKORC1-1639G>A polymorphisms was done by polymerase chain reaction-restriction fragment length polymorphism assay. A number of samples were also analyzed by direct DNA sequencing for validation of results. Data were analyzed using SPSS version 20. Genotype frequency distributions of VKORC1 1173C>T and VKORC1-1639G>A were found to be different from other populations. Both of these polymorphisms did not demonstrate significant effect on warfarin dose requirement. Although Cytochrome P450 2C9 (CYP2C9) and VKORC1 polymorphisms together attributed only 3.8% variability in warfarin dose but it was statistically significant ( p value = .004). It is concluded that there is a need to study genotype frequency distribution and their effect on warfarin dose variability among different populations due to diversity in outcome. At the same time, no effect on warfarin dose variation explained by VKORC1 polymorphisms and small variability explained by studied genotypes stresses the need for exploration of more genetic and nongenetic factors in Pakistani population.

Background Genetic information has the potential to create a more personalised, prompt, early and accurate risk evaluation. The effect of these genetic variants on the serum biomarker levels (phenotype) needs to be studied to assess their potential causal role in the pathogenesis of premature coronary artery disease (PCAD). Objectives were to determine the genotypic distribution of interleukin (IL) 18, tumour necrosis factor-α (TNFA), IL6 and IL10 single nucleotide polymorphisms (SNPs) in Pakistani PCAD cases and disease free controls and to study the effect of these gene polymorphisms on the serum cytokine levels (IL18, TNFA, IL6 and IL10) and cytokine imbalance (IL18:IL10 and TNFA:IL10). Material and methods The case–control study was carried out in National University of Sciences and Technology, Islamabad in collaboration with the Cardiovascular Genetics Institute, University College London, UK. Subjects (n=340) with >70% stenosis in at least a single major coronary artery on angiography were taken as PCAD cases along with 310 angiographically verified controls. ELISA was performed for measuring the concentrations of serum IL18, TNFA, IL6 and IL10. Genotyping was done using TAQMAN assay. Results The risk allele frequencies (RAFs) of rs1800795 (IL6) and rs187238 (IL18) cytokine gene promoter SNPs were significantly higher in the PCAD cases as compared with the controls. Serum IL18 and IL10 levels were significantly greater in the IL18 rs187238 GG genotype patients while serum IL18 and IL6 levels were significantly higher in patients having the IL6 rs1800795 CC genotype. IL18 SNP rs1946519 significantly altered the IL18, TNFA, IL6, IL18/IL10 and TNFA/IL10 ratio levels followed by TNFA SNP rs1800629 which significantly altered the serum levels of IL18, IL18:IL-0 and TNFA:IL10 ratios. Conclusions The association of the selected SNPs with differential serum cytokine levels especially the cytokine imbalance points towards their potential causal role in the immune inflammatory pathogenic pathway of PCAD.

Objectives: Interleukin-6 receptor (IL-6R) gene (A>C, rs8192284) polymorphismhas been associated with inflammatory biomarkers. We sought to investigate the association ofIL-6R gene (A>C, rs8192284) polymorphism with IL-6, IL-18 and hS-CRP levels in PCAD. StudyDesign: Case control study. Setting: Army Medical College, National University of Sciencesand Technology, Islamabad, Pakistan Methods: Total 520 subjects were recruited. 281 PCADpatients aged ≤45 years with >70% stenosis in at least one major coronary vessel along with239 age and sex matched controls were recruited. IL-6R polymorphism was determined byTaqMan genotyping while IL-6 and IL-18 levels were measured using ELISA technique and hSCRPon Immulite 2000. Results: The genotype distribution of IL-6R(rs8192284) in the cases andcontrols was: AA-56%(n=143);AC-36% (n=102); and CC-13% (n=36) and AA-59%(n=139);AC-33% (n=80); CC-8% (n=20) respectively. The risk allele frequency was significantly differentbetween the cases and controls (p=0.038) .IL-6 levels were significantly high (p

Polymorphisms in cytochrome P450 (CYP) 2C9 (CYP2C9) gene result in interindividual variability in warfarin dose requirement. There is a need for characterization of genotype frequency distribution in different populations for construction of customized dosing algorithms to enhance the efficacy and reduce the toxicity of warfarin therapy. This study was carried out in Pakistani population to evaluate the contribution of common CYP2C9 polymorphisms to warfarin therapy. A total of 550 stable patients taking warfarin were enrolled after medical history, physical examination, and laboratory investigations. Single blood sample was collected after informed consent. Genomic DNA was extracted, and genotype analysis for CYP2C9*2 and CYP2C9*3 polymorphisms was done by polymerase chain reaction-restriction fragment length polymorphism assay. A number of samples were also analyzed by direct DNA sequencing for validation of the results. Data were analyzed using SPSS version 20. Genotype frequency distribution of CYP2C9*2 and CYP2C9*3 was found to be different from other populations. Of these 2 polymorphisms, CYP2C9*2 did not demonstrate significant effect on warfarin dose requirement, whereas CYP2C9*3 did show significant effect ( P value = .012). It is concluded that there is a need to study genotype frequency distribution and their effect on warfarin dose variability among different populations due to diversity in outcome.

To determine the association of human resistin gene RETN C-420G single nucleotide polymorphism with type 2 diabetes mellitus in a specific ethnic population..The controlled study was conducted from June 2012 to January 2015 at Military Hospital, Rawalpindi, Army Medical College, Rawalpindi, and the Institute of Biomedical and Genetic Engineering, Islamabad, Pakistan. Patients with type 2 diabetes and healthy controls belonging to Pakistani Punjabi Rajput ethnic group were genotyped for human resistin gene RETNC-420G single nucleotide polymorphism. Serum resistin, serum insulin, fasting blood sugar, lipid profile, body mass index and insulin resistance was determined and correlated with genotypes. SPSS 18 was used for data analysis.Of the 789 subjects, 539(68%) were diabetics and 250(32%) were controls. Serum resistin levels were significantly higher in diabetics than controls (p0.05). The frequency of GG, GC and CC was 15(2.8%), 322(59.75%) and 202(37.5%) in diabtics. This single nucleotide polymorphism was associated with diabetes (p0.02).Human resistin gene RETN C-420G single nucleotide polymorphism was not associated with serum resistin, insulin, body mass index, insulin resistance and dyslipidaemia in both groups (p0.05 each).Human resistin gene RETN C-420G single nucleotide polymorphism was found to be a risk factor for type 2 diabetes in Pakistani Punjabi Rajput population..

Objective To compare the efficacy of albumin and C3 complement as serum marker for the early diagnosis of endometriosis in women of reproductive age. Study design Cross-sectional comparative study. Place and duration of study Biochemistry Department, Islamic International Medical College, Rawalpindi, in collaboration with the Gynecology and Obstetrics Department, Pakistan Railways Hospital, Rawalpindi, from March 2017 to February 2018. Methodology Forty-four patients with endometriosis and 44 controls of reproductive age group were enrolled for the study. Those having other comorbid systemic or endocrine diseases were excluded from the study. Levels of albumin and C3 complement were measured by using Micro Lab 300 and ELISA, respectively; and the results were compared by using SPSS version 21. groups, respectively (p=0.201). The mean serum C3 levels were 1120.9±265 μg/ml and 2241.0±293 μg/ml in the control and endometriosis patient groups, respectively (p Results The mean serum albumin levels were 4.62±0.88 g/dl and 4.42±0.52 g/dl in the control and endometriosis Conclusion: Serum C3 complement were significantly increased in endometriosis patients but not in controls; whereas, serum albumin levels were not significantly different in endometriosis cases as compared to the controls.

Hepatitis C virus (HCV) vaccines, designed to augment specific T-cell responses, have been designated as an important aspect of effective antiviral treatment. However, despite the current satisfactory progress of these vaccines, extensive past efforts largely remained unsuccessful in mediating clinically relevant anti-HCV activity in humans. In this study, we used a series of immunoinformatics approaches to propose a multiepitope vaccine against HCV by prioritizing 16 conserved epitopes from three viral proteins (i.e., NS34A, NS5A, and NS5B). The prioritised epitopes were tested for their possible antigenic combinations with each other along with linker AAY using structural modelling and epitope–epitope interactions analysis. An adjuvant (β-defensin) at the N-terminal of the construct was added to enhance the immunogenicity of the vaccine construct. Molecular dynamics (MD) simulation revealed the most stable structure of the proposed vaccine. The designed vaccine is potentially antigenic in nature and can form stable and significant interactions with Toll-like receptor 3 and Toll-like receptor 8. The proposed vaccine was also subjected to an in silico cloning approach, which confirmed its expression efficiency. These analyses suggest that the proposed vaccine can elicit specific immune responses against HCV; however, experimental validation is required to confirm the safety and immunogenicity profile of the proposed vaccine construct.

CK2 enzyme is up regulated in several cancers. It has many substratesincluding survivin which is up regulated in cancers. Objectives: To find out the correlationbetween expression of CK2α and survivin and evaluate it as a prospective prognostic marker inpathogenesis of the breast cancer and to find if positive correlation between CK2 and survivinwas associated with advancing disease. Study Design: Cross Sectional Analytical type of study.Setting: Department of Biochemistry & Molecular Biology, Army Medical College, Rawalpindiand Armed Forces Institute of Pathology, Rawalpindi. Duration of study: January 2013-December 2014. Methods: The research protocol was approved by Armed Forces Institute ofPathology Ethical Committee. Paraffin embedded tissue sections of diagnosed breast cancer,obtained from AFIP, were used. Immunohistochemistry was performed to determine nuclearand cytoplasm expression of survivin, and CK2 .Scoring done by three histopathologists,independently. Results: Total CK2 expression was high in invasive as compared to noninvasivecases (p =0.209). Cytoplasm and nuclear localization of CK2 in invasive group was alittle higher too (p = 0.092) and (p=0.286) respectively. Total survivin expression was high ininvasive as compared to non-invasive cases (p= 0.449). Cytoplasm and nuclear localizationof survivin in invasive group was higher as compared to noninvasive group with no significantdifferent (p=0.472) and (p=0.367) respectively. A positive and strong correlation was found inCK2 and survivin expression and localization in both non-invasive as well as invasive groups.Conclusion: CK2 and survivin correlation in cancers can be used in predicting the cancerphenotype and aggression at early stages.

Evaluation of a single nucleotide polymorphism (SNP) and single nucleotide mutation in cancer patients and other diseases is crucial for genotypic characterization in order to select therapy and genetic counseling. The study aim is to develop and optimization for the detection of miR-146a gene rs2910164 C/G polymorphism in breast cancer. Tetra-primer Amplification Refractory Mutation System (T-ARMS) PCR was applied on 60 individuals to determine the genotype and allele frequencies. Genotype frequencies of miR-146a gene rs2910164 C/G polymorphism were homozygous CC 65%; heterozygous CG 6.65%; homozygous GG 28.35%; allele C and G frequencies were 68.5% and 31.5% respectively. These were in Hardy–Weinberg’s equilibrium; p=0.18. Our study provides a simple rapid and a cost-effective T-ARMS-PCR molecular technique that could be used for the detection allele specific DNA polymorphisms and mutations in certain diseases and normal population at large scale.

OBJECTIVE To investigate the relation between serum amyloid A-low density lipoprotein (SAA-LDL) and genotoxicity in smokers. STUDY DESIGN An experimental study. PLACE AND DURATION OF STUDY Army Medical College, Rawalpindi and National Institute of Health (NIH), Islamabad, from June 2014 to February 2015. METHODOLOGY Seventy healthy Sprague Dawley rats were purchased from NIH and exposed to cigarette smoke in smoke chamber for three months. Blood samples were drawn from each rat at the end of the study period. SAA-LDL was determined by enzyme-linked immunosorbent assay (ELISA). Genotoxicity was assessed by cytokinesis block micronucleus (CBMN) assay. Pearson correlation was used to find correlation between SAA-LDL and genotoxicity. RESULTS Strong positive correlation was found between SAA-LDL and micronuclei frequency in smoke-exposed rats (r=0.799, N=70, p

Among solid tumors, hepatocellular carcinoma (HCC) emerges as a prototypical therapy-resistant tumor. Considering the emerging sorafenib resistance crisis in HCC, future studies are urgently required to overcome resistance. Recently noncoding RNAs (ncRNAs) have emerged as significant regulators in signalling pathways involved in cancer drug resistance and pharmacologically targeting these ncRNAs might be a novel stratagem to reverse drug resistance. In the current study, using a hybrid Petri net based computational model, we have investigated the harmonious effect of miR-17-92 cluster inhibitors/mimics and circular RNAs on sorafenib resistant HCC cells in order to explore potential resistance mechanisms and to identify putative targets for sorafenib-resistant HCC cells. An integrated model was developed that incorporates seven miRNAs belonging to miR-17-92 cluster (hsa-miR-17-5p, hsa-miR-17-3p, hsa-miR-19a, hsa-miR-19b, hsa-miR-18a, hsa-miR-20a and hsa-miR-92) and crosstalk of two signaling pathways (EGFR and IL-6) that are differentially regulated by these miRNAs. The mechanistic connection was proposed by the correlation between members belonging to miR-17-92 cluster and corresponding changes in the protein levels of their targets in HCC, specifically those targets that have verified importance in sorafenib resistance. Current findings uncovered potential pathway features, underlining the significance of developing modulators of this cluster to combat drug resistance in HCC.

Insecticide toxicity is the problem of every person in under developed countries. It is necessary to counteract its effect by natural and cheap remedies like green tea and vitamin C. In this manner common man can also enjoy blessings of life. The current research was performed to compare the protective function of green tea and vitamin C on experimental cypermethrin provoked nephrotoxicity.Forty healthy Balb/C mice purchased from National Institute of Health, Islamabad, Pakistan and divided in to four groups (10 each). Group a was control which received only normal diet. Group B, group C and group D were experimental groups which were given Cypermethrin, Cypermethrin with green tea and Cypermethrin with vitamin C respectively. These groups were also given normal diet. After 1 month blood was drawn by intra-cardiac method to assess renal parameters.One month research showed increase in serum urea to 6.8±.48 m.mol/l (n=3.9±.44) while green tea and vitamin C normalize them to 4.0±.83m.mol/l and 3.4±.33m.mol/l respectively. Serum creatinine increased to 42.90±3.28m.mol/l (n=29.50±3.95) while green tea and vitamin C normalize them to 28.80±4.58m.mol/l and 22.60±2.06m.mol/l correspondingly.The results showed that green tea and vitamin C neutralized toxicity induced by Cypermethrin in mice and their effect is comparable.

To determine the association of interleukin-6 C-174G single nucleotide polymorphism with type 2 diabetes mellitus and metabolic parameters.This case-control study was conducted from June 2012 to December 2013 at the Military Hospital Rawalpindi, the Centre for Research in Experimental and Applied Medicine, Army Medical College, Rawalpindi, and the Institute of Biomedical and Genetic Engineering, Islamabad, Pakistan. Two cohorts of subjects were genotyped for the single nucleotide polymorphism. One cohort comprised type 2 diabetics and other included healthy subjects. In these groups, serum interleukin-6, serum insulin, blood sugar fasting, lipid profile, body mass index and insulin resistance was determined and correlated with genotypes.Of the 789 participants, 539(68.3%) were in the study group and 250(31.7%) in the control group. Serum interleukin-6 was significantly higher in diabetics than healthy controls (p0.0001). The frequency of GG, GC and CC was 267(49.5%), 235(43.6%) and 37(6.9%) in diabetic patients and 128(51.2%), 74(29.6%) and 48(19.2%) in healthy controls, respectively. Interleukin-6 C-174G single nucleotide polymorphism was significantly associated with diabetes [odds ratio = 3.22 (95% confidence interval: 2.04-5.1; p0.0001). Genotypes were within Hardy-Weinberg equilibrium. Interleukin-6 C-174G single nucleotide polymorphism was significantly associated with serum interleukin-6 in the order of GCGGCC but was not associated with body mass index, insulin resistance, serum insulin and dyslipidaemia in diabetic patients (p0.05 each).Interleukin-6 C-174G single nucleotide polymorphism was a risk factor in type 2 diabetes and contributed to higher serum interleukin-6 levels among the participants.

According to World Health Organization, the prevalence of diabetes mellitus (DM) is on the rise. Tissue necrosis factor- alpha (TNF-α) is one of the inflammatory markers which play role in the pathogenesis of DM. Single nucleotide polymorphism (SNP) at 308G/A has been reported to be associated with type 2 DM. Frequency of TNF-α G308A polymorphism was determined in different ethnic groups and is found to be highly variable. Limited number of studies reported a positive association between type 2 DM and TNF-α SNP and many studies; including meta-analyses failed to find such association. TNF-α G308A polymorphism has been found to be associated with insulin resistance and BMI, although these findings are challenged by many studies. The probable explanation of higher BMI is the association of presence of TNF- α G308A polymorphism with higher rates of lipid synthesis, and suppression of FFA levels in obese persons. There is no single explanation for such highly variable results in different ethnic groups. Presence of yet unidentified gene polymorphism in linkage disequilibrium with TNF-α gene polymorphism could be responsible. The different results can also be contributed to the fact that the study groups differ in age, gender distribution, age of onset of disease, life style, degree of obesity and glucose tolerance. Comprehensive genetic studies of whole TNF-alpha promoter area are required to be done in large population samples in different ethnic groups. Polymorphisms are usually co-related in a complex manner and are co-inherited and conclusions are difficult to be drawn on small sample size.

Summary Background Autosomal recessive hypotrichosis is a rare human hereditary disorder presenting as sparse scalp hair or as woolly hair occurring on various parts of the body. Various forms of isolated hypotrichosis have been reported to date. Mutations in at least 11 genes have been reported to cause hypotrichosis. Aims To investigate the clinical and genetic basis of autosomal recessive hypotrichosis in two unrelated consanguineous families. Methods Genotyping by highly polymorphic microsatellite markers established linkage in both families to the DSG4 gene on chromosome 18q21. PCR amplification of exons and intron–exon borders of the DSG4 gene was performed, and the products sequenced to search for disease-causing sequence variants. Results Clinical investigation revealed typical hypotrichosis in the affected members of one family, while other affected members showed presence of monilethrix-like scalp hair. Sequence analysis of DSG4 revealed a novel deletion mutation (c.85-1_191del) in the affected subjects of both families. Conclusions This study further extends the body of evidence that mutations in the DSG4 gene result in both hypotrichosis and monilethrix-like scalp hair.

Background Premature Coronary Artery Disease (PCAD) occurs almost a decade earlier in the South Asian population as compared to the West. Inclusion of genetic information can prove to be a robust measure to improve early risk prediction of PCAD. Aim was to estimate the genotypic distribution and risk allele frequencies of 13 Coronary Artery Disease (CAD) risk Single Nucleotide Polymorphisms in loci identified by the CARDIoGRAMplusC4D consortium namely MIA3 rs17465637 ; 9p21 rs10757274 ; CXCL12 rs1746048 ; APOA5 rs662799 ; APOB rs1042031 ; LPA rs3798220 ; LPA 10455872 ; MRAS rs9818870 ; LPL rs328 ; SORT1 rs646776 ; PCSK9 rs11591147 ; APOE rs429358 ; APOE rs7412 in Pakistani PCAD patients and controls. Moreover, the differential serum cytokine levels (IL-18 , IL-10 , IL-6 , TNF-alpha , IL-18:IL-10 & TNF-alpha:IL-10 ratios) with respect to the genotypic distribution of these selected SNPs were determined. Material and methods The case-control study was carried out in National University of Sciences and Technology, Islamabad in collaboration with the Cardiovascular Genetics Institute, University College London, UK. Subjects (n = 340) with >70% stenosis in at least a single major coronary artery on angiography were taken as PCAD cases along with 310 angiographically verified controls. ELISA was performed for measuring the concentrations of serum IL18, TNFA, IL6 and IL10. Genotyping was done using TAQMAN and KASPar assays. Results The risk allele frequencies (RAF) of APOE rs7412, CXCL12 rs1746048, 9p21 rs10757274, MIA3 rs17465637 and SORT1 rs646776 were significantly higher in the PCAD cases as compared to the controls. APOE rs429358 had the greatest influence among the selected GWAS/CARDIoGRAMplusC4D consortium CAD risk SNPs by significantly altering the serum levels of TNF-alpha, IL-10 and TNF-alpha:IL-10 ratio. It was followed by APOE rs7412 and CXCL12 rs1746048 which significantly altered the serum levels of IL-18; TNF-alpha and IL-18; IL-18:IL-10 ratio respectively. The cytokine imbalance denoted by IL-18:IL-10 was significantly higher in the risk allele carriers MIA3 rs17465637 and CXCL12 rs1746048 while TNF-alpha:IL-10 ratio was significantly raised in the risk allele carriers of APOE rs429358 ; MRAS rs9818870 and LPL rs328 . Conclusion The association of the selected SNPs with differential serum cytokine levels especially the cytokine imbalance points towards their potential causal role in the immune inflammatory pathogenic pathway of PCAD.

Presence of different forms of ammonia in the wastewater treatment plants are toxic to aquatic life and promote eutrophication in receiving water bodies. It becomes imperative to lower the ammonia concentration in the effluents for which characterization of microbial diversity is foremost objective. Study was planned to identify the diversity of nitrifiers and denitrifiers from activated sludge using PCR technique. Cultural techniques revealed high bacterial diversity was found in the aeration tank where majority of the isolates belongs to class γ proteobacteria followed by β proteobacteria suggesting the adaptability of these groups. Presence of Nitrosomonas europaea revealed better ammonium oxidation condition at longer SRT of 6 and 7 d with DO concentration of 2.5 mg/L. As a part of NOB, Nitrospira sp. found to be more dominant than Nitrobacter winogradskyi as its detection was achieved in all samples of 6 and 7 d SRT, while later was detected only on 7 d SRT. Use of cultural techniques in combination with PCR analysis with 16S rRNA approach revealed Pseudomonas aeruginosa as important denitrifier. Highest removal efficiencies of COD, BOD, TN and NH 4 + – N were found to be 81, 78, 70 and 69% respectively at optimized SRT of 7 d.

Inflammation plays a role in the etiology of type 2 diabetes mellitus (DM). Interleukin-6 is one of the inflammatory markers which play role in the pathogenesis of diabetes mellitus DM. Single nucleotide polymorphism (SNP); -G174C in interleukin −6 (IL-6) gene promoter area has been reported to be associated with type 2 DM (T2DM). The frequency of polymorphism is found to be variable in various ethnic groups and also within an ethnic group. Many studies reported a positive association between T2DM and IL-6 SNP although quite a number of other studies failed to find such association. IL-6 polymorphism has been found to be associated with higher serum IL-6 levels, insulin resistance and BMI, although these findings are also challenged by many studies. There is no single explanation for such highly variable results in different studies. Presence of yet unidentified gene polymorphism in linkage disequilibrium with IL-6 SNP could be responsible. The different results can also be attributed to the study groups differing as age, gender distribution, age of onset of disease, life style, degree of obesity and glucose tolerance. This review highlights the varying results reported in association of IL 6 –G174C SNP with risk of T2DM, serum IL-6 levels, BMI and insulin resistance.