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Background and objectives: Hashimoto's thyroiditis is an important autoimmune thyroid condition. It is characterized by lymphocytic congestion of the thyroid gland followed by progressive deterioration and fibrous substitution of the thyroid in the parenchymal structure. This study has provided insight into the variations of blood pro-inflammatory cytokine levels in patients with Hashimoto's disease and the key role of vitamin D levels among selected patients. Materials and Methods: A total of 144 participants including healthy controls and patients were studied in the current study in which 118 were female and 26 were male. The thyroid profile was evaluated in patients with Hashimoto's thyroiditis and healthy controls. Results: The mean ± SD Free T4 in the patients was recorded as 14.0 ± 4.9 pg/mL, and TSH was 7.6 ± 2.5 IU/L, whereas the median ± IQR thyroglobulin antibodies (anti-TG) were 285 ± 142. Thyroid peroxidase antibodies (anti-TPO) were 160 ± 63.5, whereas in the healthy controls, the mean ± SD Free T4 was recorded as 17.2 ± 2.1 pg/mL, and TSH was 2.1 ± 1.4 IU/L, whereas the median ± IQR anti-TGs were 56.30 ± 46.06, and anti-TPO was 5.6 ± 5.12. The assessment of pro-inflammatory cytokines (pg/mL) and total Vitamin D levels (nmol/L) in patients with Hashimoto's thyroiditis was recorded with values IL-1B 6.2 ± 0.8, IL-6 9.4 ± 0.4, IL-8 7.5 ± 0.5, IL-10 4.3 ± 0.1, IL-12 3.8 ± 0.5, TNF-α 7.6 ± 1.1, and total vitamin D 21.89 ± 3.5, whereas in healthy controls the mean ± SD IL-1B was 0.6 ± 0.1, IL-6 2.6 ± 0.5, IL-8 3.0 ± 1.2, IL-10 3.3 ± 1.3, IL-12 3.4 ± 0.4, TNF-α 1.4 ± 0.3 and total vitamin D was 42.26 ± 5.5. Conclusions: It was found that individuals with Hashimoto's thyroiditis had raised serum levels of IL-1B, IL-6, IL-8, IL-10, IL-12, and TNF-α as compared to the healthy controls, whereas the total vitamin D levels were remarkably low as compared to health controls. Serum TSH, anti-TG, and anti-TPO levels were typically lower in controls and much higher in individuals with Hashimoto's thyroiditis. The current study's findings might aid in future studies and in the diagnosis and management of autoimmune thyroid disease. [ABSTRACT FROM AUTHOR]

Byline: Qudsia Bashir Amir Rashid Abdul Khaliq Naveed Rao Saad Ali Khan Suhail Razak and Aqeel Younis Abstract OBJECTIVE: to investigate the role of STAT1 in treatment failure in interferon [...]

Background: 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. Methods: It was an experimental study carried out at the Department of Biochemistry & Molecular 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 II & III 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. Results: 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. Conclusion: 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. [ABSTRACT FROM AUTHOR]

Warfarin is administered as a racemic preparation of R- and S-enantiomers. S-warfarin is more potent than Rwarfarin, so changes in blood levels of S-warfarin affect the anticoagulant response. This study was carried out to determine the effect of CYP2C9*2 and CYP2C9*3 polymorphisms on S/R warfarin ratio. A single blood sample was collected 12-16 hours after drug administration from 170 stable patients fulfilling the criteria. Genotyping of the CYP2C9 polymorphisms was done by polymerase chain reaction-restriction fragment length polymorphism assay. Sand R-warfarin enantiomers extraction from plasma was accomplished by a validated HPLC method. The concentration of S-warfarin was significantly different among CYP2C9 genotypes (p =0.018) whereas there was no effect on Rwarfarin (p =0.134). There was statistically significant effect of different CYP2C9 genotypes on S/R warfarin ratio (p=0.000). It is concluded that CYP2C9 polymorphisms influence CYP2C9 enzymatic activity in turn affecting Swarfarin levels but not R-warfarin, thus leading to different S/R warfarin enantiomers ratio among different CYP2C9 genotypes. [ABSTRACT FROM AUTHOR]

Objective: To investigate the genotoxic effect of cigarette smoking in Sprague Dawley rats. Study Design: Randomized experimental study. Place and Duration of Study: Study was conducted at Army Medical College Rawalpindi, from Jan 2013 to Jul 2013. Material and Methods: Seventy healthy Sprague Dawley rats were randomly divided into two groups at National Institute of Health (NIH). Group-I rats were not exposed while Group-II rats were exposed to cigarette smoke for 3 months. Cytokinesis Block Micronucleus Assay (CBMN) was done to determine genotoxicity. Data were subjected to statistical analysis using independent sample t-test. Results: The mean frequency of micronuclei in peripheral blood lymphocytes of control and smoker groups was 3.06 ± 0.639 and 6.77 ± 0.731 respectively. The difference was not statistically significant (p>0.05). Conclusion: The mean micronuclei frequency in peripheral blood lymphocytes of the smoke exposed rats was higher than that of the control rats however the difference was not significant. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of cigarette smoking on interleukin-6 in Sprague Dawley rats. Study Design: Randomized controlled study. Place and Duration of Study: Study was conducted at Army Medical College Rawalpindi, from Jan to Apr 2015. Material and Methods: Seventy healthy male Sprague Dawley rats were randomly divided into two groups at National Institute of Health (NIH). Group-I rats were not exposed while group-II rats were exposed to cigarette smoke for 3 months. Interleukin-6 (IL-6) was determined using commercially available enzyme linked immunosorbent assay (ELISA) kit. Data was analyzed using t-test. Results: Higher levels of IL-6 were observed in exposed than in non-exposed. The mean of IL-6 levels of control and smoker groups was 40.84 ± 9.09 and 49.48 ± 19.69 respectively. The difference was statistically significant (p<0.05). Conclusion: Positive association was found between cigarette smoking and IL-6. This shows that IL-6 may be used as a biochemical marker to assess inflammatory damage in smoke related diseases. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Objective: To find out whether Casein Kinase 2 (CK2) and Survivin co-express and correlate in prostate as well as in breast cancer. Study Design: A cross sectional analytical study. Place and Duration of Study: Study was done at Army Medical College and in collaboration with Armed Forces Institute of Pathology (AFIP). The duration of this research was two years. Material and Methods: The research was authorized by the Ethical Committee of AFIP. CK2 expression was determined by immunohistochemistry in paraffin embedded tissues from established patients of prostate cancer (n=30) and breast adenocarcinoma (n=30). Correlation of CK2 with Survivin was evaluated. Data were analyzed through SPSS version 20. For studying the correlation between the two proteins, pearson correlation coefficient was calculated. Data was considered at p-value =0.05. Results: An expressively strong and affirmative correlation was found among expression of total CK2 and total Survivin in invasive along with non-invasive cases of both prostate and breast cancers. A significantly strong and positive correlation was found between nuclear CK2 and Survivin expression in non-invasive cases of prostate cancer and invasive as well as non-invasive cases of breast cancer. A significantly strong and positive correlation was found only between cytoplasmic CK2 and Survivin expression in non-invasive cases of prostate cancer. Conclusion: CK2 and Survivin expressions have strong and positive correlation in both prostate and breast cancer particularly in non-invasive stage of the cancer. [ABSTRACT FROM AUTHOR]

Organic anion transporter polypeptide 1B1 (OATP1B1) encoded by (SLCO1B1) gene, an uptake transporter involved in the transport of drugs and endogenous compounds and located in hepatocyte sinusoidal membrane. Objective of study was to investigate the effects of two functionally significant SNPs (388A>G and 521T>C) and their respective genotypes of SLCO1B1 gene encoding OATP1B1 on the pharmacokinetics of atorvastatin. A total of 100 subjects divided into 6 groups as per their genotype profile were recruited. A single dose of 80mg atorvastatin was orally administered and plasma concentration measured up to 48 hours. The 388A>G and 521T>C genotypes were significantly associated with each other when compared for AUC and Cmax but exhibited no significant variations in Tmax and t1/2. 521 SNP is rather more strongly associated with altered pharmacokinetics of atorvastatin when compared with the 388 SNP, though the homozygous bi-allelic variant of 388 SNP also exhibited a fairly significant variation along with homozygous bi-allelic variant of 521 SNP. The inter-individual variation in pharmacokinetics can be explained by SLCO1B1 polymorphism. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

OBJECTIVES: To evaluate role of Interleukin-6 (-174 G/C) gene promoter polymorphism and the serum level of (IL-6) in the identification of premature coronary artery disease (PCAD). METHODS: The case-control study was carried out at Army Medical College, Rawalpindi, Pakistan, from July, 2014 to Jan, 2015 in collaboration with University College London, UK. One hundred and fifty patients, <45 years of age with greater than seventy percent blockade in at least one major coronary artery on angiography, were labeled as cases. While 150 subjects who were declared negative for coronary artery disease on coronary angiography were taken as controls. Genotyping was performed using Taqman Assay while serum IL-6 was measured using Enzyme Linked Immuno-sorbent Assay (ELISA). RESULTS: Total 364 subjects participated in the study. Mean±SD age of PCAD patients was 41±3.80 while in controls it was 36±7.8 years. Serum IL-6 levels were high in the cases (p<0.01). In IL-6 -174(G/C), polymorphism homozygous CC demonstrated significant association with occurrence of PCAD. Serum IL-6 levels showed a significant rise in CC genotype subjects when compared to GC and GG (p<0.05). The serum IL-6 levels showed significant correlation with the atherosclerotic burden (p<0.05). Sensitivity-specificity of IL-6 at cut off value of 3.7 ng/ dl was 77% and 81% respectively. CONCLUSION: IL-6 has significant diagnostic potential for PCAD with moderately high sensitivity and specificity. IL-6 (-174 G/C) polymorphism CC genotype can aid in the risk prediction of PCAD. [ABSTRACT FROM AUTHOR]

Objective: To see the relationship between C-reactive protein (CRP) and microalbuminuria in type 2 diabetic patients. Study Design: Cross sectional comparative study. Place and Duration of Study: Department of Biochemistry and Molecular Biology and Centre for Research in Experimental and Applied Medicine, Army Medical College Rawalpindi over a period of 2 years from January 2009 to December 2010. Material and Methods: Non-probability convenience sampling was done. A total of 80 (n=80) subjects were recruited in the study and were divided into four groups. Each group consisted of 20 patients. Group I consisted of normal healthy individuals, group II comprised normoalbuminuric type II diabetics, group III had microalbuminuric type II diabetics, and group IV consisted of overt albuminuric type II diabetic subjects. The albuminuria status was defined as (i) normoalbuminuria: albumin excretion rate (AER) less than 30 mg / 24h (ii) microalbuminuria: (AER) between 30-300 mg / 24h (iii) overt albuminuria: (AER) greater than 300 mg / 24h. Samples of 10 ml venous blood and 24 hour urine were collected. Plasma CRP was measured by turbidimetric method using Electa. Albuminuria was determined using Microlab semi-analyser (Randox, UK). Data were analysed using SPSS version 17 and Microsoft Excel worksheet 2010 with add on Statistical Package. Mean and standard error of mean (SEM) were used to describe numeric variables among different groups. Minimum and maximum values were also obtained to evaluate variations in the collected data. Analysis of variance (ANOVA) was applied to find out significant differences among groups. ANOVA was followed by Post Hoc Tuckey's test for multiple comparisons among groups. A p-value of less than 0.05 was considered significant and a p-value of less than 0.01 was considered highly significant. Pearson's correlation coefficient (r) was applied to find out the correlation of various parameters in different groups. A positive and negative value near 1 represented strong direct and inverse relationship respectively. Results: The mean ± SEM value of urinary albumin in group I was 27.91 ± 2.26 mg/day. In group II it was 27.37 ± 2.21 mg/day, in group III it was 220.52 ± 17.83 mg/day, in group IV it was 330.78 ± 26.75 mg/day. There was significant difference between all the groups (p<0.001). In group I, the mean ± SEM value of CRP was 1806.6 ± 183.32 ng/mL. In groups II, III and IV, the mean ± SEM value of CRP was 1987.26 ± 201.66 ng/mL, 2384.712 ± 241.99 ng/mL and 3338.60 ± 338.79 ng/mL respectively. There was significant difference between all the groups (p<0.001). Conclusion: The levels of CRP are higher in diabetics as compared to normal healthy controls and there is a progressive increase in the levels of CRP with increase in microalbuminuria in type 2 diabetics. [ABSTRACT FROM AUTHOR]