Glycated Albumin and Angiopoietin-2: Possible indicators of Diabetic Retinopathy in Type-Two Diabetes.

Objectives: To compare the levels of glycated albumin and angiopoietin-2 in Type-Two diabetics with and without diabetic retinopathy. Methods: It was a cross-sectional comparative study done at University of Health Sciences, Lahore after collecting data from recruited patients from the outpatient department of Layton Rahmatulla Benevolent Trust Eye Hospital, Lahore from 1st July, 2016 to 30th Aug., 2017. A total of 80 type two diabetics of both genders fulfilling the inclusion criteria were included and divided in two groups based on absence and presence of diabetic retinopathy. Obtained data were analyzed using IBM SPSS for Windows software (version 22). For comparison of both groups, Independent "t" Test or Mann-Whitney U tests were applied accordingly. For correlation of quantitative variables in each group, Spearman rho correlation and Pearson correlation test were applied depending upon normality of data. Results: Among 80 type-two diabetics, 42 (52.5%) patients had diabetic retinopathy and 38 (47.5%) were without diabetic retinopathy. Overall, females (62.5%) outnumbered males (37.5%). Both study group were age matched (p=0.45). Mean serum albumin in diabetic retinopathy and non-diabetic retinopathy group was 4.20 ±0.56 gm/dL and 4.43 ±0.39 gm/dL respectively (p=0.031). In diabetic retinopathy group, mean glycated albumin was 1.48 (0.63-1.76) gm/dL and median IQR in non-diabetic retinopathy was 0.52 (0.23-1.10) gm/dL (p=0.003). In diabetic retinopathy group, mean glycated albumin (percent) was 30.71±18.63% and in non-diabetic retinopathy group, the median IQR was 11.80 (5.06-27.25) (p= 0.001). The angiopoietin-2 median IQR in diabetic retinopathy group 5.70 (5.47-5.80) was significantly different (p=0.033) from diabetics without diabetic retinopathy groups 5.40 (4.97-5.60). Conclusion: Our study reported raised levels of glycated albumin (percent) and angiopoietin-2 in type-two diabetics, highlighting their possible involvement in disease and its progression.

Pathogenesis of Diabetic Cardiomyopathy and Role of miRNA.

Diabetic cardiomyopathy is characterized as abnormal function and structure of myocardium associated with diabetes irrespective of other cardiac risk factors like hypertension or coronary artery disease (CAD). The pathogenesis of DCM was not well understood in the past due to its complexity but it has been discovered recently. Various factors are found to be associated with the onset of DCM including impaired calcium handling, remodeling of extracellular matrix (ECM), increased oxidative stress, altered metabolism, mitochondrial dysfunction, and endothelial dysfunction. Micro-RNAs (miRNAs) are also found to be of great importance in the pathogenesis of DCM. Different miRNAs like miR-126, miR-24, miR-1, miR-155, miR-499, and miR-199a are found to be associated with different types of heart diseases like CAD and myocardial infarction. Studies have shown that the miRNA plays a crucial role in the development of DCM and it was found that the expression levels of different miRNAs differ in patients as compared to healthy individuals. This review focuses on the pathogenesis of DCM and various factors involved in the onset of diabetic car-diomyopathy. Moreover, the probable role of miRNA in the pathogenesis of DCM is also discussed.

Plasma somatostatin and insulin like growth factor-1 levels in women with polycystic ovary syndrome.

OBJECTIVE: To estimate plasma somatostatin and insulin like growth factor-1levels in women with polycystic ovary syndrome, and to compare it with healthy controls. METHODS: The cross-sectional comparative study was conducted at the University of Health Sciences (UHS), Lahore, Pakistan, from December 2016 to January 2018, and comprised patients of polycystic ovary syndrome selected from tertiary care hospitals of the city. A group of apparently healthy women was also raised from the local community to work as controls. Anthropometric measurements, general physical examination and fasting blood glucose levels were determined for each subject. Plasma insulin, somatostatin and insulin like growth factor-1levels were estimated using enzyme-linked immunosorbent assay. Data was collected using a predesigned questionnaire and was analysed using SPSS 20. RESULTS: Of the 80 subjects, 40(50%) were cases with a mean age of 22.63±4.47 years, and 40(50%) were controls with a mean age of 22.78±4.85 years (p>0.05). The cases had higher fasting blood glucose, insulin and insulin like growth factor-1levels (p<0.05) compared to the controls. CONCLUSIONS: Insulin resistance and lower somatostatin levels along with higher insulin like growth factor-1 levels were found in women with polycystic ovary syndrome compared to healthy women.

Altered expression of vascular endothelial growth factor, vascular endothelial growth factor receptor-1, vascular endothelial growth factor receptor-2, and Soluble Fms-like Tyrosine Kinase-1 in peripheral blood mononuclear cells from normal and preeclamptic pregnancies.

Preeclampsia (PE) is the leading cause of maternal and fetal morbidity and mortality. It complicates around 2%-10% pregnancies worldwide due to imbalance between proangiogenic and anti-angiogenic factors, leading to incomplete placentation, ischemia, and endothelial dysfunction. The study was aimed to analyze the mRNA expression of vascular endothelial growth factor (VEGF) and its receptors, i.e., VEGF receptor-1 (VEGFR-1), VEGF receptor-2 (VEGFR-2), and soluble Fms-like tyrosine kinase-1 (sFlt-1) from maternal peripheral blood mononuclear cells (PBMCs) of PE patients. This was a cross-sectional comparative study comprising 18 normotensive and 18 PE patients; the patients were further divided as early-onset preeclampsia (EOP) and late-onset preeclampsia (LOP). The expression level of VEGF, its receptors (VEGFR-1 and VEGFR-2), and sFlt-1 was investigated using real-time polymerase chain reaction. There was a significant change in the mRNA expression with a decrease in VEGF, VEGFR-1, and VEGFR-2 and an increase in sFlt-1 in PBMCs of PE and normal pregnancies (P < 0.001). sFlt-1 mRNA expression was increased by 2.95-fold in the PE group with an inverse correlation with expression of VEGFR-2 (Spearman's rho = 0.68). Based on these findings, we conclude that PE is associated with decrease in the mRNA expression of VEGF, VEGFR-1, and VEGFR-2 as compared to an increase in sFlt-1 in PBMCs.

Peroxisome proliferator activated receptor gamma 34C>G variant and anthropometric parameters in metabolic syndrome.

OBJECTIVE: To determine the frequency of 34 Cytosine >Guanine (proline 12 alanine) variant of peroxisome proliferator activated receptor gamma, and to associate it with metabolic syndrome, insulin resistance and anthropometric obesity parameters. METHODS: The cross-sectional comparative study was conducted at the University of Health Sciences, Lahore, Pakistan, from September 2016 to 2017, and comprised patients of metabolic syndrome and healthy controls. Blood pressure and anthropometric measurements of all the subjects were recorded. Fasting blood sample of 4ml was taken for biochemical parameter and deoxyribonucleic acid extraction. The frequency of genetic variant was determined by amplification refractory mutation system polymerase chain reaction. Data was analysed using SPSS 22. RESULTS: Out of 400 subjects, 200 (50%) each were patients and controls. Overall, there were 308 (77%) males and 92 (23%) females. Patients had significantly higher blood pressure, body mass index, waist circumference, waist-to-hip ratio, mid-arm circumference and triceps skinfold thickness compared to the controls (p<0.0001). Insulin resistance was also significantly higher in the patients (p<0.0001) and showed significant correlation with body mass index, waist circumference, waist-to-hip ratio, mid-arm circumference and triceps skinfold thickness (p<0.05).Waist circumference and triceps skinfold thickness were significant predictors of homeostatic model assessment for insulin resistance. Overall, the frequency of homozygous dominant genotype CC of PPAR2 34C>G was 291 (72.75%), heterozygous CG was 93 (23.25%) and homozygous recessive GG was 16 (4%).There was no significant difference in frequency of genotypes between the groups (p=0.216). However, waist circumference and body mass index were significantly lower in GG genotype compared to the CC (p=0.006 versus p=0.02). CONCLUSIONS: Waist circumference and triceps skinfold thickness were found to be the significant predictors of homeostatic model assessment for insulin resistance, while no association was found between 34 C>G variant of peroxisome proliferator activated receptor gamma and metabolic syndrome.

Plasma surfactant Protein-D levels in healthy subjects and COPD patients.

OBJECTIVE: To compare plasma surfactant protein-D levels in healthy smokers and Chronic obstructive pulmonary disease patients. METHODS: The comparative study was conducted at the University of Health Sciences, Lahore, Pakistan, from January to December 2015, and comprised chronic obstructive pulmonary disease patients and healthy smokers of either gender aged 40-80 years. Plasma surfactant protein-D levels of male and female subjects were estimated and compared with lung function and tobacco exposure. Blood samples were collected after complete history, physical examination and spirometry. Plasma levels were measured using enzyme-linked immunosorbent assay. Plasma cotinine levels were also measured for the determination of tobacco as well as biomass exposure along with pack years. SPSS 20 was used for data analysis.. RESULTS: Of the 84 subjects, there were 42(50%) patients and as many controls. Both groups had 21(50%) males and as many females. There was no significant difference in the plasma surfactant protein-D levels of males and females in the patient group compared to their counterparts in the control group (p>0.05). Females developed the disease at a younger age compared to males (p=0.04). There was no significant difference in terms of pack-years and cotinine levels between the groups (p>0.05) and lung function showed greater deterioration in the females compared to males with similar tobacco exposure (p<0.05).. CONCLUSIONS: The gender did not affect plasma surfactant protein-D levels.

Dysregulation of circulating miRNAs promotes the pathogenesis of diabetes-induced cardiomyopathy.

Diabetic Cardiomyopathy (DCM) is characterized by myocardial dysfunction caused by diabetes mellitus. After-effects of diabetic cardiomyopathy are far more lethal than non-diabetic cardiomyopathy. More than 300 million people suffer from diabetes and cardiovascular disorder which is expected to be elevated to an alarming figure of 450 million by 2030. Recent studies suggested that miRNA plays important role in the onset of diabetic cardiomyopathy. This study was designed to identify the miRNA that is responsible for the onset of diabetic cardiomyopathy using in silico and in vitro approaches. In this study, to identify the miRNA responsible for the onset of diabetic cardiomyopathy, in silico analysis was done to predict the role of these circulating miRNAs in type 2 diabetic cardiomyopathy. Shared miRNAs that are present in both diseases were selected for further analysis. Total RNA and miRNA were extracted from blood samples taken from type 2 diabetic patients as well as healthy controls to analyze the expression of important genes like AKT, VEGF, IGF, FGF1, ANGPT2 using Real-time PCR. The expression of ANGPT2 was up-regulated and AKT, VEGF, IGF, FGF1 were down-regulated in DCM patients as compared to healthy controls. The miRNA expression of miR-17 was up-regulated and miR-24, miR-150, miR-199a, miR-214, and miR-320a were down-regulated in the DCM patients as compared to healthy controls. This shows that dysregulation of target genes and miRNA may contribute towards the pathogenesis of DCM and more studies should be conducted to elucidate the role of circulating miRNAs to use them as therapeutic and diagnostic options.

Comparative gene expression analysis of Fas and related genes in preeclamptic and healthy women: A cross-sectional study.

BACKGROUND: Preeclampsia is a hypertensive disorder of pregnancy affecting about 2-10% pregnancies worldwide. mRNA expression of tumor necrosis factor alpha (TNF- α ), Fas, and FasL have been reported to be altered in placental bed in preeclamptic pregnancies. We hypothesized that the expression of these genes is also altered in peripheral blood mononuclear cells (PBMCs) in preeclampsia. OBJECTIVE: To compare the expression of Fas receptor and related genes in PBMCs of preeclamptic and normotensive pregnant women. MATERIALS AND METHODS: A cross-sectional comparative study comprising of 18 cases and 18 controls was designed. 5 ml of venous blood was drawn and collected considering aseptic measures. Buffy coat was separated by centrifugation and stored at -20°C. Favor Prep total RNA Isolation Kit (Favorgen, Taiwan) was used for RNA extraction. The mRNA expression of TNF- α , Fas, and FasL was measured by real-time polymerase chain reaction in PBMCs in preeclamptic and normal pregnancies. RESULTS: A significant increase in mRNA expression of TNF- α , Fas, and FasL (p ≤ 0.001) was observed in PBMCs of preeclamptic pregnancies compared to the control group (p ≤ 0.001). Moreover, a significant positive correlation was found between the TNF- α mRNA expression and Fas and FasL (p ≤ 0.001). CONCLUSION: The results lead to the conclusion that mRNA expression of TNF- α , Fas, and FasL in the maternal PBMCs is altered in preeclamptic pregnancies and might contribute to the pathogenesis of the disease.

Serum profile of cytokines and their genetic variants in metabolic syndrome and healthy subjects: a comparative study.

AIM: To identify genetic variants in promoter areas of IL-6 -174 G>C and TNF-α -308 G>A in metabolic syndrome (Met S) and controls and associate them with Met S and serum cytokine levels.It was a cross-sectional study, including 224 cases of Met S and 200 controls. A fasting blood sample was taken and biochemical parameters including serum glucose, insulin, lipid profile, interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) were measured. Restriction fragment length polymorphism was used to identify the genetic variants of IL-6 and TNF-α. Serum levels of IL-6 and TNF-α and insulin resistance were significantly higher in cases than the controls. IL-6 showed significant positive correlation with HOMA-IR and TNF-α. CC genotype of IL-6 was associated with the increased risk of Met S (P=0.016, OR for CC vs GC+GG = 2.33, CI: 1.15-4.71). There was no significant difference of TNF-α genotypes between the cases and the controls. Serum TNF-α and IL-6 levels were significantly higher in AA and CC genotypes of TNF-α (-308 G>A) and IL-6 (-174 G>C) as compared with the GG (P=0.00 and P=0.001). Significant correlation of IL-6 with TNF-α and insulin resistance was observed that may provide us a therapeutic target for preventing metabolic derangements from insulin resistance.

Differential Expression of Placental Growth Factor, Transforming Growth Factor-ß and Soluble Endoglin in Peripheral Mononuclear Cells in Preeclampsia.

OBJECTIVE: To measure the peripheral blood mononuclear cells (PBMCs) mRNA expression of placental growth factor (PlGF), Transforming growth factor beta (TGF-ß), and soluble Endoglin (sEng) in the blood of preeclamptic and normotensive pregnant women. STUDY DESIGN: Cross-sectional analytical study. PLACE AND DURATION OF STUDY: Department of Physiology and Cell Biology, University of Health Sciences, Lahore, from November 2016 to April 2018. METHODOLOGY: The study included 50 normotensive and 57 preeclamptic patients (18-40 years of age), all in the third trimester of pregnancy. The preeclamptic group was further divided into early-onset preeclampsia (EOP) and late-onset preeclampsia (LOP). Blood samples from patients and healthy controls were collected and mRNA expression was measured (18 patients and 18 controls) by real time PCR. Statistical analyses were done using SPSS (version 22). The values were considered significant at 0.05 level of significance. RESULTS: The PBMCs mRNA expression of PlGF, TGF- and sEng were significantly different between the preeclampsia and control group (p<0.001). A significant decrease in expression of TGF- was observed in LOP group compared to controls (p<0.001); whereas, the difference in the expression of EOP compared to controls was not significant (p=0.12). Similar to TGF-, the expression of PlGF was significantly decreased among EOP and LOP compared to controls. Detailed analysis of sEng showed significantly increased expression in both EOP and LOP as compared to healthy group (p<0.001). CONCLUSION: There is a significant difference in extra-placental expression of PlGF, and sEng in preeclampsia.

Metabolic syndrome: an update on diagnostic criteria, pathogenesis, and genetic links.

Metabolic syndrome (MetS), today a major global public health problem, is a cluster of clinical, metabolic, and biochemical abnormalities, such as central adiposity, hypertension, insulin resistance, and dyslipidemias. These MetS-related traits significantly increase the risk of type 2 diabetes mellitus, adverse cardiac events, stroke, and hepatic steatosis. The pathogenesis of MetS is multifactorial, with the interplay of environmental, nutritional, and genetic factors. Chronic low-grade inflammation together with visceral adipose tissue, adipocyte dysfunction, and insulin resistance plays a major role in the progression of the syndrome by impairing lipid and glucose homeostasis in insulin-sensitive tissues, such as the liver, muscle, and adipocytes. Adipose-derived inflammatory cytokines and non-esterified fatty acids establish the link between central obesity IR, inflammation, and atherogenesis. Various studies have reported an association between MetS and related traits with single-nucleotide polymorphisms of different susceptibility genes. Modulation of cytokine levels, pro-oxidants, and disturbed energy homeostasis, in relation to the genetic variations, is described in this review of the recent literature, which also provides updated data regarding the epidemiology, diagnostic criteria, and pathogenesis of MetS.