Heterologous COVID-19 vaccines intervention effect on reactogenicity.

To determine the change in the occurrence of short-term vaccine reactions on the use of heterologous Covid-19 booster, a single centre short-term study of two months duration was conducted. It was designed as an interventional study with registered clinical trial number # SLCTR/2022/008. It was conducted on medical students and faculty of a National university of medical sciences, Rawalpindi affiliated public sector medical college. A total of 348 individuals were administered with Ad5-nCoV vaccine and 101 with mRNA-1273 vaccine. They all had been previously vaccinated with two doses of BBIBP-CorV. BBIBP-CorV reactogenicity was considered a control group. Vaccine reactions, including pain and redness at the injection site, fever, no observed reactions at all, myalgia, feeling cold, dizziness, paraesthesia in the arm, lightheadedness, had a significant change in their frequencies in comparison to homologous vaccine (BBIBP-CorV) reactogenicity. It was concluded that mixing and matching of COVID-19 vaccines result in an increase in frequency of post-vaccine short-term reactions.

Identification and Association of Polymorphism rs2073618 of the Osteoprotegerin Gene in Type 2 Diabetics with and without Retinopathy.

OBJECTIVE: To identify and determine the association of SNP (rs2073618) of OPG gene in diabetics with and without retinopathy and in healthy controls. STUDY DESIGN: Descriptive study. Place and Duration of the Study: Department of Biochemistry and Molecular Biology, Army Medical College, Rawalpindi in collaboration with Chemical Pathology Laboratory, Pak Emirates Military Hospital, Rawalpindi and Armed Forces Institute of Ophthalmology, Rawalpindi, from June 2021 to May 2022. METHODOLOGY: Participants aged 25-70 years were inducted and divided into three equal groups. Group I consisted of diabetics with retinopathy (n = 50), group II was diabetics without retinopathy (n = 50), and group III was healthy individuals (n = 50). DNA was extracted and allele specific PCR technique was adopted using specifically designed primers. Results were analysed using the software Statistical Package for Social Sciences (SPSS) version 22.0 and online bio-informatics tool SNPstats. RESULTS: CC, CG, and GG genotypes were found to be present in 94%, 4%, and 2% in diabetics without retinopathy, 92%, 4%, and 4% in diabetics with retinopathy, respectively, and 100% presence of CC genotype only in healthy controls. C and G alleles were present in 96% and 4%, respectively, in diabetics without retinopathy, with 100% presence of only C allele in healthy subjects. The genotypic assessment using the models showed no significant association. CONCLUSION: SNP rs2073618 of OPG gene was identified in all study groups without any significant distribution or association with the development of diabetic retinopathy. The major genotype C/C was found in the majority of subjects in all groups. KEY WORDS: Allele specific PCR, Diabetic retinopathy, Single nucleotide polymorphism, Type 2 Diabetes mellitus.

Aldose reductase gene polymorphism rs752010122 and retinopathy in type 2 diabetics.

Objectives: To investigate the association of polymorphism in rs752010122 in aldose reductase gene with the pathogenesis of diabetic retinopathy, and to determine the association and allelic frequency between the variant and the disease. METHODS: The cross-sectional study was conducted from June 2021 to March 2022 at Centre for Research in Experimental and Applied Medicine (CREAM) Laboratory, Department of Biochemistry and Molecular Biology, Army Medical College, in collaboration with the Armed Forces Institute of Ophthalmology, Rawalpindi, Pakistan, and comprised blood samples from subjects of either gender aged 40-70 years. The samples were divided into group I having diabetic retinopathy patients, group II having diabetics without retinopathy, and group III having healthy controls matched for age and gender. The samples were subjected to molecular analysis. Gene sequence was downloaded from the Human Genome Database and Ensemble. Data was analysed using SPSS 22. RESULTS: Of the 150 subjects, there were 50(33.3%) in each of the 3 groups. Variants of aldose reductase rs752010122 polymorphism were significantly associated with a lower risk of diabetic retinopathy (p<0.05). An odds ratio of 1 was noted for both heterozygous and homozygous genotypes (95% confidence interval: 1). CONCLUSIONS: Aldose reductase was associated with lower risk of the disease.

Comparative effects of tocopherol and ubiquinol on arsenic induced nephrotoxicity in sprague dawley rats.

Millions of the people worldwide are drinking arsenic polluted water. The need of time is to find out the mitigation strategies to cope with this issue. To evaluate the effects of tocopherol and ubiquinol individually and collectively on arsenic induced nephrotoxicity in Sprague Dawley rats. 150 Sprague Dawley rats were divided into 5 groups randomly. Animals of group I were provided with distilled water and sterile diet pellets. All other groups were given arsenic contaminated water (5mg/L) ad libitum. Moreover, ubiquinol and tocopherol (250mg/kg each) were given to group III and IV rats respectively. Whereas, both tocopherol and ubiquinol (125mg/kg each) was given to rats of group V. After 2 weeks of intervention period, serum RFTs were evaluated on micro lab. After exposure to arsenic, animals of group II showed a significant (p<0.01) elevation of serum RFTs. Treatment with ubiquinol in group III animals and tocopherol in group IV animals reduced the levels (p<0.01) of serum RFTs in these groups. Whereas, the combined effects of both these antioxidants reversed these changes to normal values (p>0.05). Both tocopherol and ubiquinol (synergistically) are more efficient in minimizing the nephrotoxicity induced by arsenic.

Gene Expression of Glyoxalase II in Diabetic Retinopathy.

OBJECTIVE: To study the gene expression of Glyoxalase II among patients of diabetic retinopathy. STUDY DESIGN: A cross-sectional comparative study. PLACE AND DURATION OF STUDY: CREAM (Centre for Research in Experimental and Applied Medicine) and the Department of Biochemistry and Molecular Biology, Army Medical College, Rawalpindi in collaboration with Armed Forces Institute of Ophthalmology (AFIO), Rawalpindi, from November 2015 to November 2016. METHODOLOGY: Individuals were enrolled, among whom 30 were cases with diabetic retinopathy and 30 were controls without the disease. Their relevant data were collected and blood samples were drawn. Individual RNA was extracted from respective samples and cDNA was synthesised from each. Expression analysis for Glyoxalase II was done and relative quantification was done using delta delta CT method. RESULTS: A total of 60 individuals of ages 40-70 years were enrolled in the study, 30 cases and 30 controls. Among these, 34 (56.67%) were males and 26 (43.3%) were females. Mean ages were 60 ±8 years in cases and 59 ±13 years in controls. Down regulation of Glyoxalase II was observed in cases as compared to controls. CONCLUSION: Downregulation of Glyoxalase II, seen among patients of diabetic retinopathy, may indicate a failure of detoxifying system leading to accumulation of glycated end products.

Expression analysis of glyoxalase I gene among patients of diabetic retinopathy.

OBJECTIVES: To study expression of glyoxalase I in patients of diabetic retinopathy. METHODS: This cross-sectional comparative study was conducted at Centre for Research in Experimental and Applied Medicine (CREAM), Department of Biochemistry and Molecular Biology, Army Medical College, Rawalpindi in collaboration with Armed Forces Institute of Ophthalmology (AFIO) from January 2015 to November 2015. Sampling technique was non- probability purposive sampling. Total 60 subjects were enrolled in two groups. Group-I comprised 30 patients of diabetic retinopathy and Group-II of 30 normal healthy controls. Clinical and demographic data was collected and fasting venous blood samples (2 ml) were drawn. RNA was extracted and subjected to cDNA synthesis. Expression analysis for glyoxalase I was carried out and relative quantification done by double delta Ct method. RESULTS: Mean age of the patients was 61.30 ±7.06 years and mean age of controls was 59.60 ± 6.43 years. There were 17 (56.7%) males and 13 (43.3%) females in Group-I while Group-II comprised 14 (46.7%) males and 16 (53.3%) females. There was down regulation of glyoxalase I among patients of diabetic retinopathy in comparison with controls when relative gene expression was calculated. CONCLUSION: Down regulation of glyoxalase I in patients of diabetic retinopathy suggests it to be a contributory factor in the development of disease.

Thiamine deficiency and its correlation with dyslipidaemia in diabetics with microalbuminuria.

OBJECTIVE: To measure and correlate the levels of thiamine and dyslipidaemia in microalbuminuric diabetics. METHODS: Cross-sectional comparative study was conducted at the Department of Biochemistry and Molecular Biology, Army Medical College, Rawalpindi, from January 2009 to December 2010, and comprised 60 known diabetic patients, who were inducted from diabetic clinics of Rawalpindi. These patients were divided into three equal groups, with group I (n=20) being normal healthhy individuals, group II comprised of microalbuminurics type 2 diabetics (n=20) and group III (n=20) were macroalbuminuric type 2 diabetics, based on their albumin excretion rate. The healthy volunteers (n=20) had blood glucose less than 6 mmol/L and were inducted as the comparison group. Fasting blood samples of diabetic and control groups were analysed for glucose, glycosylated haemoglobin, lipid profile, thiamine chloride and thiamine monophosphate. Besides, 24-hour urine samples were analysed for microalbuminuria, thiamine chloride and thiamine monophosphate. RESULTS: Plasma thiamine chloride and thiamine monophosphate levels were found to be significantly (p<0.001) reduced in the diabetics (n=60) compared to the controls (n=20). Furthermore, there was a progressive decline in these levels with increasing albuminuria; the lowest being in the macroalbuminuric group (group IV). Urinary thiamine levels were significantly (p<0.001) higher in the diabetics compared to the controls. These changes were more pronounced as albuminuria level increased; the highest being in group IV. The parameters of lipid profile, including triglycerides, total cholesterol and low-density lipoprotein cholesterol, were significantly (p<0.001) higher in diabetics and showed progressive increase with worsening albuminuria. Whereas, the high-density lipoprotein cholesterol levels were significantly (p<0.001) reduced in diabetics and showed progressive decline as the microalbuminuria status worsened. Furthermore, a significant negative correlation was found between plasma thiamine and all the parameters of lipid profile except high-density lipoprotein cholesterol which had a significant positive correlation. A significant linear regression of microalbuminuria on plasma thiamine was also found. CONCLUSION: Thiamine levels were reduced in the diabetic population and this reduction in thiamine level was negatively correlated with lipid profile in microalbuminuric diabetics.

Levels of inflammatory markers and their correlation with dyslipidemia in diabetics.

OBJECTIVE: To measure the levels of inflammatory markers (serum ferritin and high sensitivity C-reactive protein) and dyslipidemia in diabetics and to find a correlation between these inflammatory markers and dyslipidemia. STUDY DESIGN: Comparative study. PLACE AND DURATION OF STUDY: Department of Biochemistry and Molecular Biology, Army Medical College, Rawalpindi, from March 2007 to February 2008. METHODOLOGY: The study included 30 known type-2 diabetic patients randomly inducted from diabetic clinics of Rawalpindi. Healthy volunteers (n=30) having blood glucose less than 6 mmol/L were inducted as the comparison group. Fasting blood samples of diabetics and controls were analyzed for glucose, glycated hemoglobin (HbA1c), lipid profile, high sensitivity C-reactive protein (hs-CRP) and serum ferritin. RESULTS: The diabetic subjects had significantly higher levels of glucose, HbA1c, total cholesterol, LDL cholesterol, triglycerides, hs-CRP and ferritin as compared to normal subjects (p<0.001), while the level of HDL cholesterol was significantly lower in diabetics (p<0.001). Furthermore, a significant positive correlation was found between the inflammatory markers, hs-CRP and ferritin, and the parameters of dyslipidemia i.e. total cholesterol, LDL cholesterol and triglycerides (p<0.001 r=0.72) except for HDL cholesterol, which had an insignificant negative correlation with the inflammatory markers (p>0.05 r=-0.10). CONCLUSION: Low-grade inflammation exists in Diabetes mellitus and it is positively related with dyslipidemia (except for HDL cholesterol) in diabetics.