Determination of Glyoxalase-1 levels and Identification of Genetic Variants in GLO1 Gene in Patients of Diabetic Nephropathy.

Objective: To determine the association of diabetic nephropathy with glyoxalase-1 enzyme levels and a genetic missense variation (SNP rs4746) in its gene (GLO-1). Methods: This cross-sectional comparative study was conducted at the Department of Biochemistry and Molecular Biology, Army Medical College, Rawalpindi from November 2020 to December 2022. One hundred patients and one hundred and thirteen healthy controls were enrolled using the nonprobability convenience sampling method. Medical history and 10ml blood were obtained from each individual after written informed consent. Blood samples were subjected to biochemical tests and DNA extraction which was later used for single nucleotide polymorphism (SNP) analysis (C332C variant of rs4741 GLO-1 gene) using Tetra primer ARMS PCR and gel electrophoresis. Glyoxalase-1 enzyme activity in serum was measured using ELISA. Results: There was a significant difference in serum glyoxalase-1 levels in the two groups (p-value< 0.001). The patient group had lower levels (16.24 ± 22.51mg/dl) of glyoxalase-1 as compared to the control group (48.70 ± 42.54mg/dl). In genotypic analysis, 98 out of 100 control individuals had AA genotype-while only one had CC and another AC genotype. In the patient group, 94 out of 100 patients showed AA genotype, three AC, and three CC genotypes. As the statistical significance (p-value) was 0.37, there was no significant association found between AC or CC genotype and diabetic nephropathy. Conclusion: Glyoxalase-1 levels are linked to the development of diabetic nephropathy in our patients while a known missense variant rs4746 in the GLO-1 gene is not associated with increased risk.

Frequency Of Raised Serum Lactate Dehydrogenase In Patients With Jak2 Positive Polycythaemia Vera.

BACKGROUND: Serum Lactate Dehydrogenase (LDH) is an easily available bio marker used to determine prognosis in various Myeloproliferative Neoplasms (MPN). Its utility in Polycythaemia Vera (PV) is yet to be accessed. The purpose of this study was to determine the frequency of raised serum LDH in patients with JAK2 V617F positive PV patients and its clinic-pathological association. METHODS: A cross-sectional study using non probability consecutive sampling was conducted at our institute from July 2018 to June 2019. Adult patients of either gender, newly diagnosed with JAK-2 V617F positive PV were included. Patients' demographics, clinical characteristics and baseline CBC and LDH levels were analysed. Stratification was done with regards to age, gender, diabetes mellitus, hypertension, splenomegaly and thrombosis to see the effect of these modifiers on patients with raised LDH by using Chi Square test. p-value≤0.05 was considered as significant. RESULTS: Forty patients were inducted in the study with male to female ratio of 2:1. Twenty-two (55%) patients had raised LDH levels and showed significant association with diabetes mellitus (p=0.001), splenomegaly (p=0.001) and thrombosis (p=0.018). CONCLUSIONS: This study observed raised LDH levels in almost half of JAK2 V617F positive PV patient. It warrants a larger scale study and suggests the value of plasma LDH to be used as a future prognostic marker in PV.

Frequency of Zygosity in Jak-2 Positive Patients with Polycythemia Vera-Pakistan's Perspective.

BACKGROUND: Estimation of JAK2V617F mutational load in Polycythemia Vera (PV) helps to determine the severity of the disease phenotype, the risk of thrombotic events, progression to post-PV myelofibrosis and survival. Amplification Refractory Mutation Screening (ARMS) PCR or Allele Specific (AS) PCR is a simple easy method with a reasonable sensitivity for screening of zygosity.The purpose of this study was to see the frequency of disease burden and phenotypic characteristics in Pakistani patients diagnosed with JAK2V617F mutation positive PV. MATERIALS AND METHODS: A cross-sectional study using non probability consecutive sampling was conducted at Hematology Department, Liaquat National Hospital Karachi from October 2018 to July 2019.Adult newly diagnosed JAK2V617F positive PV patients of either gender were included. Patients' demographics, clinical characteristics and baseline CBC were noted. JAK2V617F zygosity was qualitatively analyzed by ARMS-PCR technique. Age and gender were stratified to see to see the result of qualitative and quantitative effect modifiers on these patients using Chi Square and fisher exact test as appropriate while mean comparison was done by independent t-test and one way ANOVAtest. P value of ≤0.05 was considered as significant. RESULTS: Fifty one patients were included in the study with an average age of 59.60±14.29years.90.2% of patients had hypertension.All patients tested positive for heterozygous state. Significant association of gender was found with smoking (p=0.001) while age was significantly linked with hypertension (p-0.033). CONCLUSION: JAK2V617F positive PV patients are mainly heterozygous males showing significant association with smoking and hypertension. ARMS-PCR is a robust technique to determine zygosity which can be used for screening purposes.
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Outcome of heat stroke patients referred to a tertiary hospital in Pakistan: a retrospective study.

BACKGROUND: Climate change has heightened the threat of heat stroke in previously temperate zones. AIMS: This study aimed to assess the outcome of patients in relation to mortality and the role of effect modifiers among heatstroke patients presenting to a tertiary care hospital in Karachi during June 2015. METHODS: A retrospective observational study was conducted on heatstroke patients 20-27 June 2015 at the Emergency Room(ER) of a private hospital in Karachi, Pakistan. Patients' demographic data, disease severity, presentation and outcomes were determined. Statistical data was reported as numbers, percentages and mean ± SD. RESULTS: In total, 315 patients reported to ER; 76.6% patients survived, 23% expired. Males were 55% and 60% patients were fully mobile. Hypertension was the most frequent concurrent disorder. Fever documented in 79.4% and CNS derangement in 73.3% patients were the top most presenting features. Fever and disease severity were found to exert significant impact on disease outcome. Mortality rate dropped from 26 June onwards from 24.35% to 15.9% by using evaporative cooling technique combined with air conduction and maintaining room temperature at 22-24°C. CONCLUSIONS: Poor outcome during heatstroke can be minimized by advance planning and timely intervention in lowand middle-income countries.

Genotoxic And Cytotoxic Effects Of Oral Vanadyl Sulphate.

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. CONCLUSIONS: 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.

Comparison Of Biliary Stenting And Surgical Bypass In Palliative Management Of Irresectable Periampullary Carcinoma.

BACKGROUND: Some 20-40% of the periampullary carcinoma is irresectable at the time of diagnosis. Biliary stenting and surgical bypass are commonly used palliative procedure. There is no consensus favouring one procedure over the other. This study compares the both procedures. METHODS: This Randomized Controlled Trial included 47 patients who presented with diagnosis of obstructive jaundice due to periampullary carcinoma to the Department of Surgery, Federal General Hospital, Islamabad from July 2012 to December 2014. RESULTS: Out of total 47 patients 27 (57.44%) were males and 20 (42.55%) were females. Group-A included 25 (53.19%) patients while group-B included 22 (46.81%) patients. The mean age in both groups was 62.34 years (SD=±5.01). All patients died during the study. The mean survival time for the stent patients was 7.5 months while the mean survival time for surgical bypass patients was 8.3 months. The jaundice was relived in all surgical (22, 100%) of the patients as compared to (18, 72%) of the patients in stent group. CONCLUSIONS: We concluded that surgical bypass as a primary procedure in selected patients provided better jaundice relieve as compared to biliary stenting..

Effects of oral vanadium on glycaemic and lipid profile in rats.

OBJECTIVE: Vanadyl sulphate, an inorganic tetravalent salt of transition metal vanadium is conventionally used to treat diabetes and by athletes as body-building supplement. Vanadyl sulphate is a constituent of many supplements and herbal preparations available over the counter in many parts of the world. In this study the efficacy of the salt as hypoglycaemic agent and its effects on lipid profile were determined when administered in therapeutic dose range (in humans) to healthy Sprague Dawley rats for a considerable duration. METHODS: One hundred and five rats were randomly divided into three groups of 35 rats each. Animals of all three groups were provided normal rodent diet and water ad libitum. Group I animals were administered 0.5 ml plain water through oral gavage while group II and group III rats, 0.25mg/Kg/day and 1.2mg/Kg/day vanadyl sulphate respectively for 24 weeks. At the end of 24 weeks intra-cardiac blood sampling was done and blood glucose, insulin and lipid profile were measured. RESULTS: There was significant decrease in plasma glucose, insulin and HDL-c levels while LDL-c, TGs and TC levels were significantly increased in a dose dependent manner in treated groups. CONCLUSIONS: Our study showed that vanadyl sulphate possesses hypoglycaemic effect in healthy rats while insulin levels are also decreased which may be secondary to hypoglycaemia. Moreover it causes unfavorable derangement of lipid parameters in treated rats. In conclusion vanadyl sulphate though contains significant hypoglycaemic effects; its use in humans may be re-evaluated to establish its safety in relation to lipid profile.