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Objectives: To evaluate and compare the pharmacokinetic parameters, especially bioavailability, of annatto-based tocotrienol with palm tocotrienol-rich fraction in healthy human volunteers for better therapeutic outcome. Method: The systematic review was conducted between April and August 2021 in accordance with the Preferred Reporting Items for Systematic Review and Meta Analysis guidelines, and comprised search on PubMed, Google Scholar, Pakmedinet and Google search engines for open-label or double-blind randomised controlled trials involving healthy human volunteers published till January 2021. Key words used included annatto-based tocotrienol, palm tocotrienol-rich fraction, absorption and bioavailability. Boolean operators were also used, like tocotrienol AND bioavailability, annatto tocotrienol AND pharmacokinetics. Results: Of the 230 articles identified, 50(21.7%) articles met the eligibility criteria. Of them, 7(14%) were selected for data extraction and detailed analysis. Pharmacokinetic parameters of annatto-based tocotrienol were better than palm-derived tocotrienol. Oral administration of all the isomers of annatto-based tocotrienols resulted in dose-dependent increase in area under curve and plasma levels. Amongst all the isomers of annatto-based and palm-derived tocotrienol, delta isomer of annatto-based tocotrienol had the highest bioavailability with area under curve 7450±89 ng/ml, time to reach peak plasma levels 4 hours, maximum plasma concentration 1591±43 ng/nl and elimination half-life 2. 68 ±0.29 hrs. Pharmacokinetic parameters of delta isomer of annatto-based tocotrienol was greater than palm tocotrienol-rich fraction. Conclusion: Bioavailability of annatto-based tocotrienol was better than that of palm-derived tocotrienol-rich fraction. Delta isomer of annatto-based tocotrienol had the highest bioavailability amongst all isomers of tocotrienol.

Early diagnosis of coronary artery disease (CAD) in patients with chest pain is a challenge. Currently diagnosis of CAD is confirmed by coronary angiography, which is invasive and not easily available in developing countries. Therefore, it is imperative to establish noninvasive biomarker for early diagnosis of CAD in patients with angina and determine the diagnostic accuracy of inflammatory biomarkers of atherosclerosis in comparison to angiography and correlate with severity of CAD in patients with angina. Diagnostic accuracy study was carried out in tertiary care hospitals, Rawalpindi, Pakistan. Total of 58 patients aged 55.24 ± 11.61 years, with chest pain and troponin-I -ve, having coronary artery stenosis ≥50% were included as cases of CAD and 55 subjects having stenosis50% were included as controls. Nuclear factor kappa-B (NF-κB), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and high-sensitivity C-reactive protein (hs-CRP) were analyzed on immunoassay analyzers. The receiver operating characteristic curve analysis revealed significant (

Objective: To determine the frequency of disorders leading to sudden cardiac death (SCD). Study Design: Cross-sectional study. Place and duration of study: Pathology Lab in collaboration with the Cardiology Department, AFIC & NIHD, Rawalpindi Pakistan, from Jan 2017 to Dec 2018. Methodology: A total of 305 cases of sudden cardiac death reported within the last 24 hours of a death to AFIC Rawalpindi were included consecutively. Cases with a history of extracardiac diseases leading to sudden death were excluded. Most of the included cases were diagnosed phenotypically based on clinical examination, biochemistry, ECG, echocardiography, angiography, thallium scan, electro-physiological studies, cardiac CT scan, MRI and routine autopsy whenever recommended. Results: Out of 305 cases, 197 (65%) were males and 108 (35%) females. Disorders leading to SCD were found in 178 (58.3%) cases due to ischemic heart disease (IHD), aortic dissection (0.7%), hypertrophic cardiomyopathy (HCM) (0.3%), dilated cardiomyopathy (DCM) (10.5%), congenital heart disease (12.1%) and valvular heart disease (7.9%). While in cardiac channelopathies, catecholaminergic polymorphic ventricular tachycardia (CPVT) in 5 (1.6%) and congenital long QT syndrome (LQTS) in 2 (0.7%) cases. However, 24 (7.9%) cases remained as sudden unexplained deaths (SUD). Conclusion: In our setup, ischemic heart disease and dilated cardiomyopathy were the commonest causes of sudden cardiac death, followed by congenital heart disease and valvular heart diseases.

Introduction:Alpha-Cypermethrin is a synthetic pyrethroid insecticide. Pyrethroids are derived from pyrethrins, which are toxic components found in theChrysanthemum cinerariaefoliumflowers. They have been employed in agriculture, home and veterinary formulations to control the insect vectors of disease, agricultural insects and topically applied to eradicate veterinary pests. The extensive use of pyrethroids in agriculture results in severe pyrethroid poisonings in developing countries. Objectives:This study aimed to evaluate the serum biochemical changes induced by oral exposure of cypermethrin and the protective effects of selenium in female Sprague-Dawley rats. Methodology:The female rats (n=20) were equally divided into four groups. Group 1, 2, 3 and 4 received 1 ml corn oil, 55 mg/kg/b.w of cypermethrin, 1 ppm of sodium selenite, both cypermethrin and sodium selenite respectively for 21 days. The serum was utilized for estimation of alkaline phosphatase (ALP), alanine transaminase (ALT), urea, creatinine and bilirubin by using Selectra E (Germany). Levels of sodium (Na+) and potassium (K+) in serum were measured using Easylyte (Na+/K+) analyzer (Japan). Results:Results obtained showed that activities of ALP and ALT were significantly (P Conclusion:Oral exposure of cypermethrin induces injuries in liver and kidney of rats as well as alterations in the levels of serum electrolytes which were modulated by selenium co-administration.

MicroRNAs (miRNAs) play a key role in the regulation of genes for normal metabolism in the liver. Dysregulation of miRNAs is involved in the development and progression of non-alcoholic fatty liver disease (NAFLD). We aimed to explore changes in circulating miRNA expression in response to delta-tocotrienol (δT3) and alpha-tocopherol (αTF) supplementation and correlate them with relevant biochemical markers in patients with NAFLD. In total, 100 patients with NAFLD were randomized to either receive δT3 (n = 50) 300 mg or αTF (n = 50) 268 mg twice/day for 48 weeks. Plasma expression of miRNA-122, -21, -103a-2, -421, -375 and -34a were determined at baseline, 24 and 48 weeks of intervention using RT-qPCR. Both δT3 and αTF significantly downregulated expression of miRNA-122, -21, -103a-2, -421, -375 and -34a. Moreover, δT3 was more effective than αTF in reducing expression of miRNA-375 and -34a. A significant correlation was observed between miRNA expression and biochemical markers of hepatic steatosis, insulin resistance (IR), oxidative stress (OS), inflammation and apoptosis. δT3 and αTF exert hepato-protective effects by downregulating miRNAs involved in hepatic steatosis, IR, OS, inflammation and apoptosis in patients with NAFLD. Furthermore, δT3 has more pronounced effects than αTF in reducing miR-375 and miR-34a, which are linked to regulation of inflammation and apoptosis.

The Vitamin D External Quality Assessment Scheme (DEQAS) distributes human serum samples four times per year to over 1000 participants worldwide for the determination of total serum 25-hydroxyvitamin D [25(OH)D)]. These samples are stored at −40 °C prior to distribution and the participants are instructed to store the samples frozen at −20 °C or lower after receipt; however, the samples are shipped to participants at ambient conditions (i.e., no temperature control). To address the question of whether shipment at ambient conditions is sufficient for reliable performance of various 25(OH)D assays, the equivalence of DEQAS human serum samples shipped under frozen and ambient conditions was assessed. As part of a Vitamin D Standardization Program (VDSP) commutability study, two sets of the same nine DEQAS samples were shipped to participants at ambient temperature and frozen on dry ice. Twenty-eight laboratories participated in this study and provided 34 sets of results for the measurement of 25(OH)D using 20 ligand binding assays and 14 liquid chromatography–tandem mass spectrometry (LC–MS/MS) methods. Equivalence of the assay response for the frozen versus ambient DEQAS samples for each assay was evaluated using multi-level modeling, paired t-tests including a false discovery rate (FDR) approach, and ordinary least squares linear regression analysis of frozen versus ambient results. Using the paired t-test and confirmed by FDR testing, differences in the results for the ambient and frozen samples were found to be statistically significant at p

Purpose:Cypermethrin (class II) belongs to pyrethroid insecticides which have been in wide use to control various pests including moth pests of cotton, fruit and vegetable crops. This study was aimed to investigate the effects of cypermethrin on haematology of female rats and their modulation by co-administration of sodium selenite. Methodology:Four equal groups of female rats were made (n=20). Group 1 received 1 ml corn oil, Group 2 was administered 55 mg/kg/b.w of cypermethrin, Group 3 was provided with 1 ppm of sodium selenite and Group 4 was given both cypermethrin and sodium selenite. Haematological analysis of blood samples was performed employing Sesmex Kx-21 (Japan). Results:Red blood cell (RBC) count, haemoglobin concentration (Hb) and hematocrit (Hct) were significantly reduced (P Conclusion:Hence, it can be concluded from this study that cypermethrin administration can cause hematological alterations in rats. Co-administration of selenium provided protection against cypermethrin toxicity to some extent.

An interlaboratory comparison study was conducted by the Vitamin D Standardization Program (VDSP) to assess the performance of ligand binding assays (Part 2) for the determination of serum total 25-hydroxyvitamin D [25(OH)D]. Fifty single-donor samples were assigned target values for concentrations of 25-hydroxyvitamin D(2) [25(OH)D(2)], 25-hydroxyvitamin D(3) [25(OH)D(3)], 3-epi-25-hydroxyvitamin D(3) [3-epi-25(OH)D(3)], and 24R,25-dihydroxyvitamin D(3) [24R,25(OH)(2)D(3)] using isotope dilution liquid chromatography – tandem mass spectrometry (ID LC-MS/MS). VDSP Intercomparison Study 2 Part 2 includes results from 17 laboratories using 32 ligand binding assays. Assay performance was evaluated using mean % bias compared to the assigned target values and using linear regression analysis of the test assay mean results and the target values. Only 50% of the ligand binding assays achieved the VDSP criterion of mean % bias ≤ |±5%|. For the 13 unique ligand binding assays evaluated in this study, only 4 assays were consistently within ±5% mean bias and 4 assays were consistently outside ±5% mean bias regardless of the laboratory performing the assay. Based on multivariable regression analysis using the concentrations of individual vitamin D metabolites in the 50 single-donor samples, most assays underestimate 25(OH)D(2) and several assays (Abbott, bioMérieux, DiaSorin, IDS-EIA, and IDS-iSYS) may have cross-reactivity from 24R,25(OH)(2)D(3). The results of this interlaboratory study represent the most comprehensive comparison of 25(OH)D ligand binding assays published to date and is the only study to assess the impact of 24R,25(OH)(2)D(3) content using results from a reference measurement procedure.

An interlaboratory study was conducted through the Vitamin D Standardization Program (VDSP) to assess commutability of Standard Reference Materials(®) (SRMs) and proficiency testing/external quality assessment (PT/EQA) samples for determination of serum total 25-hydroxyvitamin D [25(OH)D] using ligand binding assays and liquid chromatography – tandem mass spectrometry (LC-MS/MS). A set of 50 single-donor serum samples were assigned target values for 25-hydroxyvitamin D(2) [25(OH)D(2)] and 25-hydroxyvitamin D(3) [25(OH)D(3)] using Reference Measurement Procedures (RMPs). SRM and PT/EQA samples evaluated included SRM 972a (four levels), SRM 2973, six College of American Pathologists (CAP) Accuracy-Based Vitamin D (ABVD) samples, and nine Vitamin D External Quality Assessment Scheme (DEQAS) samples. Results were received from 28 different laboratories using 20 ligand binding assays and 14 LC-MS/MS methods. Using the test assay results for total serum 25(OH)D [i.e., the sum of 25(OH)D(2) and 25(OH)D(3)] determined for the single-donor samples and the RMP target values, the linear regression and 95% prediction intervals (PIs) were calculated. Using a subset of 42 samples that had concentrations of 25(OH)D(2) below 30 nmol/L, one or more of the SRM and PT/EQA samples with high concentrations of 25(OH)D(2) were deemed non-commutable using 5 of 11 unique ligand binding assays. SRM 972a (Level 4), which has high exogenous concentration of 3-epi-25(OH)D(3) was deemed non-commutable for 50% of the LC-MS/MS assays.

The study aimed to ascertain the effects of delta-tocotrienol (δT3) supplementation on glycemic control, oxidative stress, inflammation and related micro-ribonucleic acid (miRNA) expression in patients with type 2 diabetes mellitus (T2DM). Total 110 patients of T2DM on oral hypoglycemic agents, were randomly divided into tocotrienol and placebo groups and given 250 mg δT3 or cellulose soft gel capsule once daily respectively for 24 weeks. Glycemic control, oxidative stress, inflammatory biomarkers, and miRNAs expression were measured in serum at baseline and end of the intervention by using standard laboratory methods. Compared to the placebo, δT3 supplementation resulted in a significant (p ≤ .05) reduction [mean difference (95% confidence interval)] in plasma glucose [-0.48 (-0.65, -0.30)], insulin [-1.19 (-1.51, -0.87)], homeostatic model assessment of insulin resistance [-0.67 (-0.86, -0.49)], glycosylated hemoglobin [-0.53 (-0.79, -0.28)], malondialdehyde [-0.34 (-0.45, -0.22)], high sensitive-C-reactive protein[-0.35 (-0.54, -0.16)], tumor necrosis factor-alpha [-1.22 (-1.62, -0.83)], and interleukin-6[-2.30 (-2.91, -1.68)]. More than twofold downregulation in miRNA-375, miRNA-34a, miRNA-21, and upregulation in miRNA-126, miRNA-132 expression was observed in the δT3 group compared to the placebo. The study demonstrated that δT3 supplementation in addition to oral hypoglycemic agents, improved glycemic control, inflammation, oxidative stress, and miRNA expression in T2DM without any adverse effect. Thus, δT3 might be considered as an effective dietary supplement to prevent long-term diabetic complications.

The Vitamin D External Quality Assessment Scheme (DEQAS) distributes human serum samples four times per year to over 1000 participants worldwide for the determination of total serum 25-hydroxyvitamin D [25(OH)D)]. These samples are stored at -40 °C prior to distribution and the participants are instructed to store the samples frozen at -20 °C or lower after receipt; however, the samples are shipped to participants at ambient conditions (i.e., no temperature control). To address the question of whether shipment at ambient conditions is sufficient for reliable performance of various 25(OH)D assays, the equivalence of DEQAS human serum samples shipped under frozen and ambient conditions was assessed. As part of a Vitamin D Standardization Program (VDSP) commutability study, two sets of the same nine DEQAS samples were shipped to participants at ambient temperature and frozen on dry ice. Twenty-eight laboratories participated in this study and provided 34 sets of results for the measurement of 25(OH)D using 20 ligand binding assays and 14 liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. Equivalence of the assay response for the frozen versus ambient DEQAS samples for each assay was evaluated using multi-level modeling, paired t-tests including a false discovery rate (FDR) approach, and ordinary least squares linear regression analysis of frozen versus ambient results. Using the paired t-test and confirmed by FDR testing, differences in the results for the ambient and frozen samples were found to be statistically significant at p 0.05 for four assays (DiaSorin, DIAsource, Siemens, and SNIBE prototype). For all 14 LC-MS/MS assays, the differences in the results for the ambient- and frozen-shipped samples were not found to be significant at p 0.05 indicating that these analytes were stable during shipment at ambient conditions. Even though assay results have been shown to vary considerably among different 25(OH)D assays in other studies, the results of this study also indicate that sample handling/transport conditions may influence 25(OH)D assay response for several assays.

Objective: To study the effects of delta-tocotrienol on glycaemic control parameters in individuals with pre-diabetes. Method: The randomised control trial was conducted at the Armed Forces Institute of Pathology, Rawalpindi, Pakistan, from July 15 to November 15, 2019, and comprised individuals aged 18-60 years having fasting plasma glucose of 5.6 to 6.9mmol/L or glycosylated haemoglobin of 5.7 to 6.4%. They were randomised into group A receiving 300mg delta-tocotrienol and group B receiving a placebo once daily for 12 weeks. Weight, height, waist circumference, fasting plasma glucose, insulin and glycosylated haemoglobin were measured at the beginning and end of the trial to assess any change. Body mass index and homeostatic model assessment-insulin resistance were also calculated. Data was analysed using SPSS 21. Results: Of the 77participants, 40(52%) were in group A and 37(48%) in group B. Group A showed significantly greater reduction in terms of fasting plasma glucose, glycosylated haemoglobin, insulin and homeostatic model assessment-insulin resistance index (p?0.001) post-intervention. Conclusion: Delta-tocotrienol supplementation was found to have a significant effect in improving glycaemic control parameters in persons with pre-diabetes. Futures larger scale clinical trials are needed to confirm these findings. Continuous...

OBJECTIVE To find out the use of screening and confirmatory tests for diagnosis of multiple myeloma as ordered by clinicians. STUDY DESIGN An Audit. PLACE AND DURATION OF STUDY Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Rawalpindi, from January 2012 to January 2017. METHODOLOGY Data retrieved from Laboratory Information Management system (LIMs) by selecting serum protein electrophoresis (SPE) as screening test and immunoelectophoresis (IE) and immunofixation (IF) as confirmatory tests. RESULTS There were 3,108 tests of serum protein electrophoesis and 1,329 tests of immunoelectophoresis had been performed in last five year. Cost-effective clinical audit of SPE tests showed that only 17.1% tests of SPE were either used for proper diagnosis of multiple myeloma (totally justified tests 13%) or useful for diagnosis of other important diseases whose early diagnosis were helpful for patients management (partially justified tests 4.1%). The cost related to these tests were justified while 82.8% tests of SPE were either normal (total unjustified tests 24.4%), or diagnosed irrevalent and nonspecific diseases (partially unjustified 58.4%). IE and IF audit revealed that only 26.6% tests were properly utilised for diagnosis and differentiation of multiple myeloma and its subtypes and cost attributable to these tests were justified while 73.4% of these confirmatory tests were normal and cost related to them was not justified. CONCLUSION Overutilisation of laboratory tests for diagnosis of multiple myeloma can be minimised by proper clinical scrutiny of request forms.

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

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

The aim of this study was to examine the effects of delta-tocotrienol (δ-tocotrienol) supplementation on biochemical markers of hepatocellular injury and steatosis in patients with nonalcoholic fatty liver disease (NAFLD).The study design was a two-group, randomized, double-blind, placebo-controlled trial. The patients with NAFLD were randomly assigned to receive δ-tocotrienol 300 mg twice daily or placebo for 24 weeks.The primary endpoints were change from baseline in fatty liver index (FLI) and homeostasis model of insulin resistance (HOMA-IR) after 24 weeks. Secondary endpoints included change from baseline in high sensitivity C-reactive protein (hs-CRP), malondialdehyde (MDA), alanine transaminase (ALT), aspartate transaminase (AST) and grading of hepatic steatosis on ultrasound. Between-group differences were tested for significance using ANCOVA. Mean differences (MD) with 95 % CIs are reported.A total of 71 patients (tocotrienol=35, placebo=36) were randomized and included in the intention to treat analysis. Compared with placebo, δ-tocotrienol significantly reduced (MD [95 % CI]) FLI (-8.52 [-10.7, -6.3]; p0.001); HOMA-IR (-0.37 [-0.53, -0.21]; p0.001), hs-CRP (-0.61[-0.81, -0.42]; p0.001), MDA (-0.91 [-1.20, -0.63]; p0.001), ALT (-8.86 [-11.5, -6.2]; p0.001) and AST (-6.6 [-10.0, -3.08]; p0.001). Hepatic steatosis was also reduced by a significantly greater extent with tocotrienol than with placebo (p =0.047). No adverse events were reported.δ-tocotrienol effectively improved biochemical markers of hepatocellular injury and steatosis in patients with NAFLD. δ-tocotrienol supplementation might be considered as a therapeutic option in the management of patients with NAFLD.Sri Lankan Clinical Trials Registry (SLCTR/2015/023, 2015-10-03).

Introduction: Genetic information which is specific to an individual has thepotential to improve Coronary Artery Disease (CAD) risk prediction. 13 CAD risk SNPs wereselected by removing SNPs in loci which had not been identified in CARDIoGRAMplusC4DGWAS. Linkage disequilibrium patterns differ between ethnic groups pointing towards theneed to investigate how the gene score would perform in different populations which is stilllargely unknown. Objective of the study was to investigate whether the 13 SNP CAD risk genescore has a role in the risk prediction of Pakistani Premature Coronary Artery Disease (CAD)cases and controls and to compare the CAD risk allele frequency between Pakistanis andCaucasians (samples obtained from the Northwick Park Heart Study II). Study Design: Casecontrolstudy. Setting: Army Medical College, National University of Sciences and Technology(NUST) in collaboration with the Cardiovascular Genetics Institute, University College London,UK. Materials and Methods: Total of 650 subjects with a history of chest pain were selected bynon-probability convenience sampling. Out of these subjects with > 70% stenosis in at least 1coronary vessel on angiography were labelled as Premature coronary Artery disease (PCAD)cases (n=340). The 13 SNPs were genotyped in a Pakistani case-control study (n=340 CADcases, 310 controls) using KASPar and Taqman assays. The use of 13 SNP gene score wastested in the prospective Northwick Park Heart Study (NPHSII) of 2775 healthy UK men (284cases) and the Pakistani case-control study subjects (n=650). Results: Mean ± SD age ofCAD patients was 42.7±3.80yrs while in controls it was 39.0±7.8yrs. Complete genotypingwas obtained for 635 samples (333 cases, 302 controls). The mean 13 SNP gene score wassignificantly higher in cases compared to controls (p=0.044). Odds ratio for CAD for eachquintile of 13 SNPs gene score showed a trend for higher quintiles of gene score to haveincreased odds ratio for CAD (p-value for trend=0.01) especially after adjusting for age, sexand ethnicity. There was a significant difference in risk allele frequency between Pakistanis andCaucasians (NPHSII) for all CAD risk SNPs except rs599839 (SORT1) (p=0.08). Conclusion:A 13 SNP gene score has significant potential role at differentiating between Pakistani PCADcases and controls. Risk allele frequencies for CAD differ significantly between Pakistanis andCaucasians stressing the need to develop population specific gene score keeping in view theethnic stratification.

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.

To determine the mean hemoglobin HbA1C levels of disease-free adults in a local population and its optimum cutoff for the diagnosis of diabetes.Cross-sectional study.Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Rawalpindi, from January to September 2015.Healthy subjects aged 18 years and above of either gender were recruited from local population. Pregnant ladies and individuals with known diabetes, chronic kidney disease, chronic liver disease, congestive cardiac failure, anemia, hemoglobinopathies, mental illness and individuals on glucocorticoid therapy were excluded. Fasting plasma glucose (FPG) or 2-hour plasma glucose (2-h PG) was analyzed using hexokinase methodology and glycated hemoglobin (Hb A1C) was also analyzed using turbidimetric inhibition immunoassay technique. Receiver operating characteristic (ROC) curves were plotted. Differences among the groups were tested by one-way ANOVA, and p0.05 was considered statistically significant.Among 558 subjects, 88.8% (496) were normoglycaemic (NG), 5.7% (32) were with impaired glucose fasting (IFG), and 5.4% (30) were diagnosed with diabetes mellitus (DM). A1C was 5.00 ±0.44% in NG and 6.28 ±1.16% in diabetics. FPG in NG was 4.55 ±0.95 mmol/Land in diabetics was 8.28 ±1.78 mmol/L. The optimal HbA1C cutoff value for diagnosis of DM was at 6.05% (AUC 0.827 95% CI 0.732 to 0.923, p ≤0.05 with its sensitivity of 53.3% and specificity of 98.5%. However, HbA1C showed suboptimal sensitivity and specificity for prediabetes.The mean HbAIC and cutoff point for diabetes in the study population is 5.07 ±0.58% and 6.05%, respectively (AUC 0.827, 95% CI: 0.732 to 0.923, p0.001) with 53.3% sensitivity and 98.5% specificity.

Cardiomyopathy is the leading cause of mortality in beta-thalassaemia major (BTM) patients. Brain natriuretic peptide (BNP) has been used for latent cardiac dysfunction in heart patients other than thalassaemic patients. The objective was to determine its role in subclinical detection of iron-induced cardiotoxicity in BTM patients.EDTA plasma was taken from 33 thalassaemic patients and 29 healthy controls, stored at -20 degrees C until analysis. The median (range) age of thalassaemic major children was 10 (6-21) years and mean serum ferritin levels were 3956 (1929-6979) microg/L. The BTM children had significantly (P0.01) higher BNP median (range) 83.94 (45.93-196.80) pg/mL as compared to controls 55.62 (32.58-99.84) pg/mL. Mitral E-wave velocities were found higher in patients rather than controls (132.12 +/- 29.40 vs. 117.70 +/- 24.81; P0.05). The E/Ea ratio was significantly higher in BTM patients than in the control group (16.35 +/- 6.01 vs. 19.26 +/- 4.67; P = 0.001). We found a significant positive correlation between BNP and E/Ea ratio (r = 0.53; P0.01). BNP at a cut-off value of 84.39 pg/mL was highly accurate in ruling out diastolic dysfunction (E/Ea8) with a sensitivity of 80% and a specificity of 88%.BNP has a good predictive value in detecting latent LV dysfunction in BTM patients.

Chromium is widely used in the leather industry, and tannery workers are under constant threat of adverse health effects due to its excessive exposure. Our objective was to find out the toxic effects of chromium on tannery workers at Sialkot, Pakistan. A total of 240 males consisting of 120 workers from tanneries at Sialkot and equal number of controls were included. Blood complete counts, high-sensitive C-reactive protein, malondialdehyde and routine biochemical tests were carried out by routine procedures. Chromium levels in blood (BCr) and urine were analyzed using graphite furnace atomic absorption spectrophotometer Perkin Elmer analyst-200. Results revealed that all the workers were male with average age of 33 years and 15 (13%) had skin rashes, 14 (12%) had chronic bronchitis, 10 (8%) had gastritis and 4 (3%) conjunctivitis. The tannery workers had significantly raised median (interquartile range) of BCr 569 (377–726) nmol/L as compared to 318 (245–397) nmol/L in the control ( p

Background We aimed to elucidate the association between gamma glutamyl transferase (GGT) activity with prevalence of premature coronary artery disease (CAD) in young Pakistani patients undergoing diagnostic coronary angiography. Methods A total of 218 young adults (age≤45 years) underwent diagnostic angiography. Serum samples were taken from all the patients and analyzed for serum GGT activity, cholesterol and triglycerides. Results Coronary artery disease patients had significantly increased GGT activity ( P =.001) and exhibited a significant positive correlation with blood pressure, cholesterol, blood glucose, and smoking and negative correlation with total antioxidant status ( P Conclusion The study revealed good diagnostic accuracy at cutoff of 35 U/L with a sensitivity of 92%, specificity of 81%, and diagnostic odds ratio of 48 in estimation of premature CAD in young Pakistanis.

Background: Early diagnosis is the cornerstone of management of acute myocardial infarction (AMI). We aimed to compare the diagnostic accuracy of high-sensitivity troponin T (hs-cTnT) with myeloperoxidase (MPO) and pregnancy-associated plasma protein A (PAPP-A) for early diagnosis of AMI in patients at the time of presentation to the emergency department (ED). Methods: We enrolled 289 patients who presented at the ED of the National Institute of Heart Disease (NIHD) Rawalpindi, Pakistan, within 4 hr of onset of chest pain. Clinical assessment, electrocardiography (ECG), and angiography were carried out. Blood samples were collected at 0, 3, 6, and 12 hr. Analyses of plasma hs-cTnT, MPO, and PAPP-A were carried out using commercial kits. Results: Out of 289 subjects who presented to the ED, we diagnosed 180 patients with coronary heart disease as having AMI (N = 61) and 119 as without AMI (stable coronary artery disease, N = 61; unstable angina, N = 58). Compared to non-AMI patients, the patients with AMI had significantly higher levels (represented here as median [inter quartile range]) of plasma hs-cTnT (136 [39-370] vs. 12 [7-21] ng/L), MPO (906 [564-1,631] vs. 786 [351-1,299] pmol/L) and PAPP-A (5.78 [2.67-13.4] vs. 2.8 [1.8-4.9] mIU/L). Receiver operator characteristic curves (95% CI) for hs-cTnT (0.952 [0.909-0.978]) were significantly higher ( P < 0.001) than those for MPO (0.886 [0.830-0.929]) and PAPP-A (0.797 [0.730-0.854]), with AMI sensitivity and specificity percentages of 87% and 98% (hs-cTnT), 82% and 84% (MPO), and 65% and 87% (PAPP-A), respectively. Conclusions: The diagnostic performance of hs-cTnT was superior to that of MPO and PAPP-A for early triage and diagnosis of AMI among patients of coronary heart disease presenting with chest pain to the ED.

Lead poisoning is a well recognized environmental health problem in children. Independent association of iron deficiency and lead exposure with elevated blood lead level (BLL) has been reported. Whether iron deficiency in combination with chronic lead exposure increases BLL and susceptibility to its harmful effects in children needs to be elucidated.In this case-control study, 246 children were randomly recruited. They comprised 123 children of lead smelters/battery recycle plant workers living close to the industries at Wah/Gujranwala, Pakistan (lead exposed group) and 123 children living 30 km away from the industrial area (controls). Blood lead analysis was carried out on the anodic stripping voltammeter lead analyzer 3010B. Blood counting was done on a Sysmex hematological analyzer and serum ferritin was determined by kit method on Immulite-1000.Of the 123 children in each group, 42 (34%) were iron deficient in the exposed group while 35 (28%) in the controls. The children's median age was 4 years (69 males and 54 females in each group). Lead exposed iron deficient children had significantly higher BLL median (quartile) 13.1 μg/dL (10.1-16.8) as compared with 9.6 μg/dL (7.6-10.3) in the iron deficient controls (P0.05). Elevated BLL level was found in 31% of the lead exposed children and in 11% of the controls. Lead exposed children revealed a stronger negative correlation (r= -0.54; P=0.001) between BLL and serum ferritin than the controls (r=-0.36; P=0.01).Iron deficiency in combination with lead exposure synergistically elevates blood lead levels and susceptibility to its harmful effects in children.

The study aimed to determine the hazardous health effects of pesticides exposure in the factory workers by measuring plasma cholinesterase (PChE), pesticides residues, and renal and hepatic biochemical markers. In addition, we also assessed the knowledge, attitudes, and safety practices adopted by the industrial workers. The study was conducted in three different sizes of factories located in Lahore (large), Multan (medium), and Karachi (small) in Pakistan. Total 238 adult males consisting of 184 pesticide industrial workers (exposed group) from large-sized (67), medium-sized (61), small-sized (56) industrial formulation factories, and 54 controls (unexposed) were included in the study. All the participants were male of aged 18 to 58 years. PChE levels were estimated by Ellmann’s method. Plasma pesticides residue analysis was performed by using reverse phase C-18 on high-performance liquid chromatograph and GC with NPD detector. Plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine, urea, and gamma glutamyltransferase (GGT) were measured on Selectra E auto analyzer. Plasma and C-reactive protein was analyzed by Immulite 1000. The results revealed a significant decrease in plasma post exposure PChE levels (

Lead is a persistent toxic metal and associated with impairment of various body functions in occupational workers. The main objective was to determine the lead-induced oxidative stress and adverse health effects by biochemical markers in industrial workers. One hundred and forty-eight males consisting of 87 lead-exposed industrial workers and 61 controls were included. Blood lead level (BLL) was determined on a 3010B ESA lead analyzer. Blood complete counts were done on a hematology analyzer. Biochemical markers including serum uric acid, urea, creatinine, phosphate, alanine aminotransferase (ALT), and gamma glutamyltransferase (GGT) were measured on a Selectra E auto analyzer. Serum malondialdehyde (MDA) was measured spectrophotometrically and C-reactive protein (CRP) on Immulite-1000. Results revealed that lead-exposed workers had significantly high BLLs, median (range), 29.1 (9.0-61.1) microg/dL compared with controls, 8.3 (1.0-21.7) microg/dL. Oxidative stress (MDA, GGT) and inflammatory markers (high-sensitivity CRP) were significantly increased (P < or = 0.05). Blood pressure was raised, whereas hemoglobin was decreased in exposed group (P < or = 0.002). Serum urea, uric acid, phosphate, and ALT were significantly raised in lead-exposed workers (P < or = 0.001). Serum albumin, total proteins, and glomerular filtration rate (GFR) were decreased. Blood lead showed a significant positive correlation with serum GGT (r = 0.63), MDA (r = 0.71), CRP (r = 0.75), urea (r = 0.34), creatinine (r = 0.51), and uric acid (r = 0.29) (P < or = 0.01). It is concluded that lead exposure increases oxidative stress that correlates with adverse changes in hematological, renal, and hepatic function in the occupational workers. Elevated blood lead has positive correlation with oxidative stress, inflammatory and biochemical markers that might be used to detect impairment in the body function in lead exposed workers.

To determine the serum 25-hydroxycalciferol levels [25(OH)D] in adults with pre-diabetes and normoglycaemia to examine a possible association of vitamin D deficiency with pre-diabetes.Case control study.Armed Forces Institute of Pathology, Rawalpindi, from November 2012 to July 2013.Atotal of 272 adults including 136 pre-diabetics and 136 normoglycaemics of either gender aged 20 years and above were consecutively inducted. Patients with diabetes mellitus, pregnancy, rickets and osteomalacia, ischemic heart disease, chronic kidney disease and chronic liver disease were excluded. Fasting Plasma Glucose (FPG) was estimated with hexokinase method on Modular p800 Roche chemistry analyzer while serum 25(OH)D was measured on Diasorin Liaison immunoassay analyzer using the chemiluminescent technique. Mean 25(OH)D levels in pre-diabetic and normoglycaemic groups were compared using Mann-Whitney U test. Spearman's correlation coefficient 'rs' was determined between serum 25(OH)D and FPG. Odds ratio for vitamin D deficiency was also calculated.Mean serum 25(OH)D level was low in pre-diabetics (23.2 nmol/L) as compared to normoglycaemics (29 nmol/L; p=0.001). Serum 25(OH)D level had inverse correlation with FPG (rs= -0.448, p=0.000). There was also significant association of vitamin D deficiency with pre-diabetes compared with normoglycaemia (OR: 2.21, p= 0.016; 95% CI: 1.15-4.27).Vitamin D deficiency with pre-diabetes suggested that vitamin D may have an important role in pathogenesis of pre-diabetes.

This study evaluated pesticide effects on reproductive and thyroid hormones of cotton farmers of southern Punjab, Pakistan. A total of 88 cotton farmers (42 spray applicators and 46 cotton pickers) were randomly included with an equal number of age- and sex-matched controls. Sampling was done in high spraying and peak picking seasons. Serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, prolactin, thyroid-stimulating hormone (TSH), total triiodothyroxine (TT3), and free thyroxine (fT4) were carried out by enzymatic immunoassay. Plasma cholinesterase (PChE) levels were measured by Ellman’s method. Serum FSH, LH, and testosterone levels were significantly high in spray applicators ( P < .01).Serum FSH and testosterone levels were significantly raised in cotton pickers ( P < .01). Serum prolactin was decreased significantly in both groups ( P < .01).Serum fT4 was significantly reduced in cotton pickers ( P < .01). Pesticide exposure is associated with thyroid and reproductive hormone levels disturbance.

To derive the ethnic factor and validate the modified estimated Glomerular Filtration Rate (eGFR) by Modification of Diet in Renal Disease (MDRD) equation for Chronic Kidney Disease (CKD) patients of Rawalpindi.Cross- sectional study.Armed Forces Institute of Pathology (AFIP), Rawalpindi, from July 2011 to July 2012.A total of 140 patients with CKD reporting to AFIP for GFR measurement by 99mTechnetium diethylenethiaminepenta-acetic acid (99mTc-DTPA) renal scan were consecutively inducted. Serum creatinine was measured by the Jaffe's assay on Beckman DxC 600 Analyzer prior to the renal scan. Ethnic factor for population of Rawalpindi with CKD was derived for the MDRD eGFR equation using 99mTc-DTPA renal scan by Gates method as the reference method. MDRD equation was modified by inclusion of the ethnic factor in it. Agreement between the reference GFR (rGFR) and the modified MDRD eGFR (mGFR) was assessed by applying paired samples t-test.Out of 140 patients of CKD, 99 (71%) were males and 41 (29%) females, with mean age of 55 ± 13.42 years. The mean values were 32.91 ± 14.96, 34.89 ± 16.45, 0.971 ± 0.20 and 33.87 ± 15.97 for rGFR, original eGFR, ethnic factor and mGFR respectively. The mGFR with new ethnic factor of 0.971 showed improved performance as compared to original eGFR and showed a significant level of correlation with rGFR (r2 = 0.817), at a p-value of 0.000.This study validates the mGFR equation by inclusion of newly derived ethnic factor of 0.971 in the population of Rawalpindi with CKD and it was found to be not significantly different from the rGFR.

To compare the Friedewald and modified Friedewald formulae with direct homogeneous assay for serum lowdensity lipoprotein cholesterol (LDL-C) levels estimation.Cross-sectional study.Armed Forces Institute of Pathology, Rawalpindi, from June to December 2011.Healthy subjects of either gender, from Rawalpindi, aged 18-75 years were included by consecutive sampling. Patients with diabetes mellitus, chronic liver disease, chronic kidney disease, those taking lipid lowering drugs and samples with triglyceride (TG)4.52 mmol/l were excluded from the study. Total cholesterol, high-density lipoprotein cholesterol, TG and LDL-C were measured on Hitachi 912 chemistry analyzer (Roche). LDL-C levels were also calculated by Friedewald formula (FF) and Vujovic modified formula (VMF). Paired sample t-test and scatter plots were used for statistical analysis.Although both calculated methods showed good correlation with direct assay (r0.93) in 300 subjects, but the difference was statistically significant. The ffLDL-C were 0.12 ± 31 mmol/l (p0.001) lower and vmfLDL-C were 0.11 ± 26 mmol/l (p0.001) higher than dLDL-C. The difference was not significant between ffLDL-C and dLDL-C at TG levels1.70 mmol/l (p = 0.58) and between vmfLDL-C and dLDL-C at TG levels 2.26 - 4.52 mmol/l (p = 0.38). At all other TG levels, the difference between LDL-C calculated by both formulas and dLDL-C was statistically significant (p0.001). As compared to direct assay, 11% and 14% subjects were classified in wrong National Cholesterol Education Programm's cardiac risk categories by FF and VMF respectively.LDL-C should be measured by direct homogeneous assay in routine clinical laboratories, as the calculated methods did not have a uniform performance for LDL-C estimation at different TG levels.

To determine the accuracy of neutrophil gelatinase-associated lipocalin (NGAL) in early detection of acute kidney injury (AKI) after cardiopulmonary bypass (CPB) surgery by comparing with serum creatinine.Descriptive study.Department of Chemical Pathology and Endocrinology, AFIP in collaboration with AFIC/ NIHD, Rawalpindi, from April to December 2011.Eighty eight patients undergoing CPB surgery in AFIC/NIHD were included by consecutive sampling. Blood samples of subjects for serum creatinine analysis were drawn pre-operatively, 4 h, 24 h and 48 h after CPB surgery. Spot urine samples for NGAL were collected at 4 h after CPB surgery. Urine samples were analyzed on Abbott ARCHITECT i2000SR analyzer whereas serum creatinine samples were measured on Beckman UniCel® DxC 600 Synchron® Clinical System.Out of 88 patients, 11 (13%) cases developed AKI 4 h postoperatively. Urinary NGAL increased markedly at 4 h postoperatively as compared to serum creatinine which showed rise at 24 - 48 h after cardiac surgery. Analysis of urine NGAL at a cutoff value of 87 ng/ml showed area under the curve of 0.91 [95% confidence interval (CI) 0.83 - 0.96] with sensitivity of 90.9% (95% CI 58.7 - 98.5) and specificity of 98.7% (95% CI 92.9-99.8). There was a positive correlation of 4 h urine NGAL and serum delta creatinine at 48 h, which was statistically significant (rs = 0.33, p = 0.001).The study demonstrated that levels of urine NGAL in patients suffering from AKI increased significantly at 4 h as compared to serum creatinine levels. Urine NGAL is an early predictive biomarker of AKI after CPB.

Anaemia in pregnancy is a common clinical problem contributing to increased maternal and foetal morbidity. This study was carried out to determine frequency of iron, folate and cobalamin deficiency and associated risk factors in the anaemic pregnant females who reported first time during second and third trimester for antenatal check-up in the tertiary care hospital at Rawalpindi.This case control study was carried out in a tertiary care hospital at Rawalpindi. Two hundred and fifty pregnant women (age: 19-43 years) consisting of 125 anaemic (Hb110 g/L) and 125 non-anaemic who reported first time at antenatal clinic were included. Data on socio-demographic characteristics, parity and dietary intake were collected. Complete blood counts were done. Serum ferritin, folate and cobalamin assays were performed by using DPC kits on Immulite-1000.The pregnant women were categorised having mild (Hb up to 54%), moderate (Hb up to 36%), or severe (Hb up to 10%) anaemia during antennal visit. They had significantly lower median (range) levels of haemoglobin 96 (40-110) g/L, ferritin 8 (3-142) microg/L, folate 15 (3-54) etamol/L and cobalamin 171 (111-629) etamolL than controls (p =0.01). Micronutrient analysis revealed secondary pregnancy related deficiency of Iron (57%), folate (20%). combined iron and folate (19%) and cobalamin (4%) in the female Among the risk factors, low income (OR: 7.69), multiparty (OR: 2.93), lack of iron/folate supplementation (OR 2.91) and inadequate dietary intakes (OR 2.51) were associated with anaemia.The pregnant anaemic women had iron (57%); folate (20%), followed by combined iron folate (19%) and cobalamin (4%) deficiency during first antenatal visit. Low income, multiparty, poor diet and lack of supplements are the main contributor in development of anaemia during pregnancy.

To determine the frequency of adverse effects attributed to Methotrexate (MTX) toxicity and serum minimum toxic concentration with low dose MTX in Rheumatoid Arthritis (RA) patients.Cross-sectional observational study.Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Rawalpindi, from March 2010 to March 2011.One hundred and forty adult patients of RA receiving low dose MTX (10 mg/week) for at least 3 months, ere included by consecutive sampling. Blood samples were collected 2 hours after the oral dose of MTX. Serum alanine transaminase and creatinine were analyzed on Hitachi and blood counts on Sysmex analyzer. Serum MTX concentration was measured on TDX analyzer.Out of one hundred and forty patients; 68 males (49%) and 72 females (51%), 38 developed MTX toxicity (27%), comprising of hepatotoxicity in 12 (8.6%), nephrotoxicity in 3 (2.1%), anaemia in 8 (5.7%), leucopenia in 2 (1.4%), thrombocytopenia in 3 (2.1%), pancytopenia in 2 (1.4%), gastrointestinal adverse effects in 5 (3.6%) and mucocutaneous problems in 3 (2.1%). Receiver operating characteristic curve revealed serum minimum toxic concentration of MTX at cutoff value of 0.71 μmol/l with a sensitivity of 71% and specificity of 76%.Adverse effects of low dose MTX were found in 27% of RA patients, mainly comprising of hepatotoxicity and haematological problems. MTX toxicity can be detected by therapeutic drug monitoring of serum concentration of 0.71 μmol/l with sensitivity of 71% and specificity of 76% in the patients on low dose MTX maintenance therapy.

Proinflammatory interleukin-18 (IL-18), high-sensitivity C-reactive protein (hS-CRP), tumor necrosis factor-alpha (TNF-alpha), and anti-inflammatory IL-10 are involved in the pathogenesis of atherosclerosis. The aim of this study was to determine the imbalance between pro- and anti-inflammatory cytokines in premature coronary artery disease (PCAD) and their association with the degree of angiographic atherosclerotic blockade. A case-control study was carried out at the National Institute of Heart Diseases, Rawalpindi, Pakistan. Three hundred eighty-five stable coronary artery disease patients aged45 years were screened. A total of 172 subjects participated in this study, comprising 98 PCAD patients and 74 angionegative controls. Serum IL-10, IL-18, and TNF-alpha were measured using enzyme-linked immunosorbent assay and hS-CRP was analyzed using Immulite 1000. The mean age of the patients was 40 ± 4.23 years (69 men and 9 women). Serum IL-18, TNF-alpha, hS-CRP, and IL-18/IL-10 ratio were significantly raised in PCAD patients (P0.01), whereas IL-10 was moderately increased (P0.05) when compared with controls. Serum IL-18 and hS-CRP were significantly raised in patients with 86%-100% stenosis (n=57) when compared with patients with 71%-85% stenosis (n=41) (P0.05). Serum IL-18, TNF-alpha, IL-18/IL-10 ratio, and hS-CRP significantly correlated (P0.01) with the degree of angiographic blockade. Pro/anti-inflammatory cytokines play a vital role in the pathogenesis of PCAD and have potential to identify the degree of atherosclerosis.

To compare pregnancy-associated plasma protein-A (PAPP-A) levels in individuals with and without coronary artery disease (CAD).Cross-sectional comparative study.Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology (AFIP), Rawalpindi, in collaboration with Armed Forces Institute of Cardiology (AFIC), from September 2008 to March 2010.One hundred and twenty five (125) individuals both male and female were included in the study. Blood for PAPP-A and lipid profile was collected, just before angiography. On the basis of angiography, the individuals were divided into those with and without CAD. PAPP-A was analyzed by using Diagnostic System Laboratories (DSL) Enzyme Linked Immunosorbent Assay (ELISA) kit and reading was taken by ELISA reader. Lipid profile was determined on automated analyzers Selectra-2 and Vitros 5.1.Amongst the 125 individuals, 41 individuals were without CAD whereas 84 individuals were having CAD. Mean PAPP-A levels were 0.74 ± 0.35 mIU/L in those without CAD whereas mean PAPP-A levels in those with CAD were 1.35 ± 0.57 mIU/L. The difference between the two groups was statistically significant (p0.001). A PAPP-A cut off level of 0.85 mIU/L had a sensitivity and specificity of 78% and 70% respectively for diagnosing atherosclerotic CAD.PAPP-A is a potentially relevant marker of the presence and extent of coronary atherosclerosis as its levels are elevated in CAD as compared to individuals without CAD.

Lead exposure is an important environmental health problem particularly affecting the children of occupational workers living in the lead-contaminated environment. The objectives of the study were to find out the frequency, potential sources and adverse health effects of elevated blood lead level (BLL) in the children of lead-related occupational workers. It was a comparative cross-sectional study. A total of two hundred forty six children aged 1—6 years, comprising an equal number (n = 123) from lead smelters/battery recycle plant workers living close to the industries at Wah/Gujranwala, Pakistan (lead-exposed group) and those living 30 km away from the industrial area (controls) were included. Demographic and clinical data of each subject was collected. Blood lead analysis was carried out by using kits on the lead analyzer (3010 B ESA, USA). Biochemical tests of renal and hepatic profile were analyzed on Selectra E auto analyzer. The median age of children was 4 years; comprising of 69 boys and 54 girls. The lead-exposed children had significantly high BLLs median (range) 8.1 (1—20.9) μg/dL as compared to controls 6.7 (1—13.3) μg/dL (p ≤ 0.01). The children of occupational workers had elevated BLL (>10 μg/dL) in 38 (31%) as compared with 14 (11%) in controls. Hematopoietic, renal, and hepatic functions were significantly impaired in the lead-exposed children. In conclusion, the children of lead-related occupational workers have significantly increased frequency (31%) of lead poisoning. The potential source of lead overexposure in these children may be indirect through father’s clothes and contaminated environment at home. Increased lead accumulation adversely affects health of these children.

Glycaemic control is critical for managing diabetes and related complications. Considering high prevalence of Diabetes in Pakistan, our study aimed to assess the status of glycaemic control in Type-II Diabetics by measurement of HbA1c from 2005-2007 at Rawalpindi. We also evaluated changes in its trends in relation with sex and age.It was a retrospective analysis of data from Clinical Pathology Laboratory, Army Medical College, Rawalpindi during 2005-2007. A total of 2875 patients, aged 24-70 years, taking oral hypoglycaemic agents, were included. HbA1c was measured by using Human kit. International Diabetes Federation guidelines,or = 6.5%, 6.6%-8.4% andor = 8.5% were used to classify patients into good, fair and poor control categories.The number of patients (n = 2875) tested for HbA1c increased from 904,974 to 997 during 2005-2007. The patients had an age of 48 +/- 13 years and comprised of 54% males and 46% females. Improvement in patient's glycaemic control among the three categories during 2005 to 2007 was as follows: good (41% vs 47%), fair (38% vs 40%) and poor (21% vs 13%) respectively. The average HbA1c values improved from 7.25 % in 2005 to 6.69% in 2007 (p0.05). Overall, males (45%) and youngest age group (53%) patients had good diabetic control.Glycaemic control improved in diabetic patients from 41% to 47% during 2005-2007 at Rawalpindi. Males, especially the youngest patients comprised majority of good control population. For effective disease management and optimal HbA1c values, a combined effort by the patient and physician is required.