Urinary megalin levels in patients with type 2 diabetic nephropathy and its correlation with renal function.

Purpose: Megalin is a glycoprotein molecule found on proximal renal tubular epithelial cells. The objectives of this study were to determine urinary megalin levels in non-diabetic subjects and in patients with and without type 2 diabetic nephropathy and to assess the correlation between urinary megalin, urinary albumin, and estimated glomerular filtration rate (eGFR) in diabetic patients. Materials and Methods: This was a cross-sectional comparative study conducted at a tertiary care teaching hospital in South India for 2 years. Study subjects were divided into three groups: non-diabetic subjects, diabetics with normoalbuminuria, and diabetics with microalbuminuria. Urinary albumin was detected by the dipstick technique in a spot urine sample for all study subjects. Nephelometry was used to quantify urinary albumin levels. The enzyme-linked immunosorbent assay technique estimated urinary megalin. Results: Urinary megalin levels were higher in non-diabetic subjects compared to diabetic study subjects. There was a significant difference in urinary megalin levels between non-diabetic subjects and diabetic patients with microalbuminuria. No correlation was found between urinary megalin, urinary albumin, and eGFR in patients with diabetic nephropathy. Conclusion: Urinary megalin levels were higher in non-diabetic subjects than in type 2 diabetic patients. There was no correlation between urinary megalin, urinary albumin, and eGFR in patients with diabetic nephropathy.

Association of elevated extracellular HSP72 in albuminuria with systemic inflammation and disease progression in type 2 diabetic kidney disease.

BACKGROUND: Sub-clinical inflammation in hyperglycemia is tied to the pathogenesis of diabetic kidney disease (DKD). Though well known for its immunostimulatory function, the significance of extracellular heat shock protein 72 (eHSP72) in DKD is not well studied. We aimed to determine the association of extracellular HSP72 with systemic inflammation and the progression of DKD, and explore its possible clinical significance in DKD. METHODS: 160 type 2 diabetic individuals were enrolled in the study. Their anthropometric data, routine biochemical parameters, urinary renal function parameters, and blood count parameters were estimated. Plasma from patients' blood samples were used to estimate HSP72 and interleukin 1ß (IL-1ß) using sandwich immunoassays. RESULTS: Plasma eHSP72 is elevated in DKD. Pairwise comparisons showed the drastic elevation of eHSP72 in the presence of albuminuria. A significant positive relationship was observed between plasma levels of eHSP72 and IL-1ß. eHSP72 levels did not statistically differ between micro and macro-albuminuric DKD. However, it was inversely associated with estimated glomerular filtration rate, the index of disease severity, independent of age, gender, diabetes duration and absolute monocyte count. At a cutoff of 0.52 ng/ml, with sensitivity of 64.1 % and specificity of 69.2 %, plasma eHSP72 differentiated the presence of DKD in type 2 diabetics with statistical significance. CONCLUSION: The positive relationship of eHSP72 and IL-1ß with worsening DKD likely indicates their participation in immunostimulatory pathways of renal fibrosis. eHSP72 may be closely linked to albuminuria-induced tubular injury and likely contributes to fibrotic changes in the progression of DKD. From our study, we infer the possible clinical significance of eHSP72 as a marker of sub-clinical renal damage in DKD, and the implication of IL-1ß-associated mechanisms in DKD progression.

Circulating 18-Glycosyl Hydrolase Protein Chitiotriosidase-1 is Associated with Renal Dysfunction and Systemic Inflammation in Diabetic Kidney Disease.

Introduction: Chitotriosidase-1 (CHIT-1) is a marker of macrophage activation and recently attributed to type 2 diabetes mellitus (T2DM). However, its role in the development and progression of diabetic kidney disease (DKD) has been sparsely discussed in the recent literature. Materials and Methods: In this cross-sectional exploratory study, 81 participants with T2DM were classified into two groups based on the presence of DKD. Their anthropometric, biochemical, and pathological profiles were estimated. Circulatory CHIT-1 concentration was determined using the enzyme-linked immuno-sorbent assay (ELISA) in plasma. Results: CHIT-1 was significantly elevated in diabetic nephropathy, independent of age and gender. It is associated with severity of kidney disease, as assessed using urinary protein-creatinine ratio (uPCR) in a multiple linear regression model, independent of age, gender, diabetes duration, and insulin resistance. CHIT-1 positively predicted the likelihood of DKD in the study population (area under the curve = 0.724, P < 0.05). The duration of diabetes correlated positively with uPCR and negatively with estimated glomerular-filtration rate. Neutrophil-Lymphocyte ratio was elevated in participants with DKD. This well-established marker of systemic inflammation exhibited significant positive association with CHIT-1. Conclusion: Plasma CHIT-1 protein is elevated in DKD and associated with disease progression. It is capable of reflecting disease severity and is closely related to systemic inflammation possibly caused by pro-inflammatory circulatory immune cells.

Nutri-stress, mitochondrial dysfunction, and insulin resistance-role of heat shock proteins.

Excess nutrient flux into the cellular energy system results in a scenario of cellular metabolic stress in diseases involving insulin resistance, such as type 2 diabetes, referred to as nutri-stress and results in cellular bioenergetic imbalance, which leads to insulin resistance and disease. Under nutri-stress, the heat shock response system is compromised due to metabolic abnormalities that disturb energy homeostasis. Heat shock proteins (HSPs) are the chief protectors of intracellular homeostasis during stress. Heat shock response (HSR) impairment contributes to several metabolic pathways that aggravate chronic hyperglycaemia and insulin resistance, highlighting a central role in disease pathogenesis. This article discusses the role of nutri-stress-related molecular events in causing insulin resistance and the nature of the roles played by heat shock proteins in some of the crucial checkpoints of the molecular networks involved in insulin resistance. Ample evidence suggests that the heat shock machinery regulates critical pathways in mitochondrial function and energy metabolism and that cellular energy status highly influences it. Weakening of HSPs, therefore, leads to loss of their vital cytoprotective functions, propagating nutri-stress in the system. Further research into the mechanistic roles of HSPs in metabolic homeostasis will help widen our understanding of lifestyle diseases, their onset, and complications. These inducible proteins may be crucial to attenuating lifestyle risk factors and disease management.

Serum Ferritin for Predicting Outcome in Children With Severe Sepsis in the Pediatric Intensive Care Unit.

OBJECTIVES: To evaluate the prognostic ability of serum ferritin when estimated within 5 days of onset of illness in children with severe sepsis admitted to a pediatric intensive care unit. METHODS: This observational study enrolled children aged 1 month to 12 years with severe sepsis. Hemoglobin, serum ferritin and C-reactive protein levels were measured within five days of illness. Final outcomes were recorded in all enrolled children. RESULTS: 70 children with median (IQR) age of 27 (8,108) months were enrolled during the study period (July, 2019 to August, 2021). 28 (40%) of these had poor outcome (non-survival). The median (IQR) level of serum ferritin was 1369 (558-5607) ng/mL in non-survivors and 282 (129-680) ng/mL in survivors (P<0.05). A significant correlation was seen between serum ferritin and Pediatric Risk of Mortality III (PRISM III) score (r=0.364 P=0.002) and pediatric Sequential Organ Failure Assessment (pSOFA) score (r=0.246 P=0.04) at 48 hours of admission. 54 (77.1%) children were anemic. Serum ferritin levels in children with anemia also had a good predictive ability for poor outcome [AUC: 0.764, 95% CI: 0.634, 0.894]. CONCLUSIONS: Serum ferritin levels, within five days of onset of illness, predicted poor outcome in critically ill children with severe sepsis and in children with microcytic anemia.

Association of serum omentin-1, apelin and chemerin concentrations with the presence and severity of diabetic retinopathy in type 2 diabetes mellitus patients.

Omentin-1 is a novel adipokine with anti-inflammatory functions. Apelin is associated with hyperinsulinemia and pathological angiogenesis. Chemerin has both pro- and anti-inflammatory actions and implicated in insulin resistance and metabolic syndrome. The aim of this study was to assess serum omentin-1, apelin and chemerin concentrations and to investigate their association with the presence and severity of DR in T2DM patients. Serum omentin-1, apelin and chemerin were measured in 112 patients with DR and 56 patients without DR. Bivariate analysis showed omentin-1 correlated negatively with hsCRP and TyG index; while apelin correlated positively with chemerin. Linear regression data showed that apelin and chemerin were independent predictors of DR severity. ROC curve revealed that omentin-1 was the best discriminant for DR while apelin was the best discriminant for vision threatening retinopathy. Serum omentin-1 concentration correlates negatively, while serum apelin and chemerin concentrations correlate positively with DR presence and severity in T2DM patients.

Incidence, Risk Factors, the Role of Plasma NGAL and Outcome of Contrast-Induced Acute Kidney Injury in Critically Ill Children.

OBJECTIVES: To study the incidence of contrast-induced acute kidney injury (CI-AKI), evaluate its risk factors, study the role of plasma neutrophil gelatinase-associated lipocalin (NGAL) and evaluate the outcome of CI-AKI in critically ill children. METHODS: In this prospective cohort study, children aged 1 mo to 12 y who underwent contrast computed tomography (CECT) for various medical indications were included. Patients without renal function test before contrast administration, children with chronic kidney disease, children admitted for less than 48 h, and those with serum bilirubin more than 5 mg per dL were excluded. Serum creatinine and estimated-Glomerular filtration rate (e-GFR) were measured at admission, immediately before, and at 6, 24, 48 h after contrast. Plasma neutrophil gelatinase-associated lipocalin (NGAL) was measured before and 6 h after contrast. The incidence of CI-AKI by p-RIFLE (Pediatric Risk, Injury, Failure, Loss, End Stage Renal Disease) criteria, its risk factors, the diagnostic role of NGAL in CI-AKI, and outcomes [30 d unfavorable outcome (death, readmission) and renal recovery] were studied. RESULTS: One hundred children were enrolled. The indications for CECT were brain (58%) and respiratory pathology (20%). Incidence of CI-AKI was 35% (95% CI 26.4% to 44.8%); 71% in 'Risk,' and 29% in the 'Injury' stage. After multivariate logistic regression, age younger than 2 y was independently associated with CI-AKI. There was no significant difference in NGAL before (ROC-AUC 0.38, 95% CI 0.26 to 0.50) and 6 h after CECT scan (AUC 0.41, 95% CI 0.29 to 0.54) to predict CI-AKI. There were 7% deaths but no readmission at 30 d. Among 33 CI-AKI patients who survived, the operational definition of renal recovery was achieved in 51.5% (n = 17), complete renal recovery was achieved in 97% (n = 32), and partial renal recovery was achieved in 3% (n = 1) of patients at discharge, while none received renal supportive therapy. CONCLUSIONS: The incidence of contrast-induced acute kidney injury was 35% with age younger than two year being independently associated with CI-AKI. NGAL did not predict the CI-AKI.

Circulating adropin and vascular endothelial growth factor receptor-2 levels in age-related macular degeneration and T2DM patients-A cross-sectional study.

BACKGROUND: Macular drusen formation and angiogenesis are the two chief processes associated with age-related macular degeneration. Adropin and vascular endothelial growth factor receptor-2 (VEGFR-2) may be involved in these pathologies. By altering lipid metabolism, adropin may contribute in the early stages of age-related macular degeneration (AMD). VEGFR-2 may participate in the later form of AMD, by promoting angiogenesis. This study compared the circulatory levels of adropin and VEGFR-2 in AMD and patients without AMD and assessed their association with disease severity, to understand their possible role in AMD. OBJECTIVES: This study aimed to assess and compare the serum levels of adropin and VEGFR-2 in patients with AMD and type 2 diabetes patients without AMD, and, to investigate the correlation between these two parameters with disease severity. METHODS: Our study involves two groups of 39 each. Group A (age-related macular degeneration) and Group B (diabetes patients without age-related macular degeneration). Routine parameters fasting blood sugar (FBS), lipid profile, and liver function tests (LFT) were estimated by using autoanalyzer. Serum adropin and VEGFR-2 were assessed by ELISA. RESULTS: Among the basic parameters, systolic blood pressure and fasting blood glucose alone were significantly different across the groups. We did not find significant alterations in adropin and VEGFR-2 levels between the study groups. Our lipid profile parameters (triglycerides and total cholesterol) have significant positive association. VEGFR-2 showed a positive correlation with the severity of AMD. Adropin did not exhibit any correlation with disease severity and with VEGFR-2. CONCLUSION: We could not find any observable alterations of statistical significance, in adropin and VEGFR-2 levels. VEGFR-2's correlation with disease severity could be important. Adropin might have subtler roles in AMD, though not evident from our study, and requires a deeper observation at the molecular level to elucidate its function.

Cardiovascular autonomic neuropathy in patients with type 2 diabetes mellitus and its association with serum omentin and leptin.

Cardiovascular autonomic neuropathy (CAN) is a major cause of morbidity and mortality in patients with diabetes as it is associated with a high risk of cardiac arrhythmias. OBJECTIVES: This prospective observational cross-sectional study was done to estimate the prevalence of CAN in patients with type 2 diabetes and to study its association with serum omentin and leptin levels. METHODS: This study included 100 patients with type 2 diabetes mellitus attending the outpatient department of JIPMER Hospital, Pondicherry, India, from January 2017 to December 2018. CAN was assessed in all subjects using four cardiovascular autonomic function tests. Blood samples were collected and stored at - 80°C to estimate leptin and omentin levels. Comparison of leptin and omentin levels was done between diabetic patients with and without CAN. RESULTS: CAN was present in 64% of the study subjects. Serum leptin levels were significantly higher in patients with CAN, whereas omentin levels, though elevated in those with CAN, were not statistically significant in diabetic patients without CAN. CONCLUSION: There is a high prevalence of CAN in patients with type 2 diabetes mellitus. Leptin levels were elevated in these patients, whereas omentin levels were not significantly different between diabetic patients with and without CAN.

Burden of diabetic kidney disease among persons with type 2 diabetes mellitus registered in a tertiary care center, Puducherry.

BACKGROUND & AIMS: To determine burden of diabetic kidney disease (DKD) and estimate yield and number needed to screen (NNS) in a tertiary diabetes care center. METHODS: DKD was diagnosed if estimated Glomerular Filtration Rate (eGFR) < 60 ml/min/1.73 m2 or Urinary Albumin Creatinine Ratio (UACR) ≥30 mg/g in two urine samples. RESULTS: Of 511 participants, 206 (40%) had DKD. Using both UACR and eGFR, yield and NNS were 39% and three, respectively. CONCLUSION: Using eGFR alone, yield and NNS to find a case of DKD among consecutive adult (≥18 years age) patients with type 2 diabetes mellitus were 26.4% and 4, respectively.

Circulating matrix modulators (MMP-9 and TIMP-1) and their association with severity of diabetic retinopathy.

AIMS: Diabetic Retinopathy (DR) is the leading cause of vision loss in the working age population. Matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1), are molecules involved in extracellular tissue matrix remodelling. They are implicated in the loss of retinal tissue integrity, a major cause of DR, that leads to retinal tissue degradation and apoptosis. This study is therefore, conducted to compare the serum levels of MMP-9 and TIMP-1 in T2DM patients without and with retinopathy, and to evaluate their association with the severity of DR. MATERIALS AND METHODS: Our study comprised of 2 groups of 41 each. Group A (cases) included T2DM patients with retinopathy and Group B (controls) included T2DM patients without retinopathy. Routine parameters, mainly, fasting blood glucose, and lipid profile were measured using autoanalyzer. Serum MMP-9, TIMP-1, and insulin levels were assessed using ELISA method. RESULTS AND CONCLUSION: Statistically significant increase in the levels of MMP-9, insulin, fasting blood glucose and lipid profile were observed in the serum of T2DM patients with retinopathy, as compared with those without retinopathy. These results help to conclude that rise in MMP-9, and associated serum markers promote disease progress in DR. These findings suggest that the elevations of our study markers in the serum of the type 2 diabetic patients with retinopathy, as compared to those without retinopathy, play important roles in aggravating tissue matrix degradation, supporting DR disease progression.

Utility of urinary biomarkers as a diagnostic tool for early diabetic nephropathy in patients with type 2 diabetes mellitus.

INTRODUCTION: Renal tubulo-interstitial damage has an important role in the pathogenesis of early diabetic nephropathy. Urinary biomarkers can help in the detection of early nephropathy in type 2 diabetic patients. The aim of this study was to estimate the levels of urinary neutrophil gelatinase associated lipocalin (NGAL) and cystatin-C in type 2 diabetic patients with early diabetic nephropathy & to compare them with diabetic patients without nephropathy and to correlate urinary NGAL and cystatin-C levels with microalbuminuria in them. STUDY DESIGN: Cross-sectional comparative study. MATERIAL AND METHODS: The study was conducted on 126 patients with type 2 diabetes along with 30 control subjects attending the outpatient care department of a tertiary care teaching hospital. There were 3 study groups-diabetic patients with microalbuminuria, diabetic patients without albuminuria and control subjects who were non-diabetic without any renal disease. Details on duration of diabetes and glycemic status were obtained from the patients. Urine examination was done for subjects in all the groups to look for microalbuminuria along with estimation of NGAL and cystatin-C levels. Samples were stored at -20 °C in the deep freezer. RESULTS: Urinary NGAL and cystatin-C levels were significantly elevated in patients with microalbuminuria (228.18 & 3.23 ng/ml) as compared to those without albuminuria (146.12 & 2.61 ng/ml) and in control subjects (26.56 & 0.30 ng/ml). Urinary NGAL and cystatin-C levels showed a linear correlation with microalbuminuria in diabetic patients. CONCLUSION: Urinary NGAL and cystatin-C levels were increased in type 2 diabetic patients with early diabetic nephropathy as compared to patients without nephropathy. Urine NGAL and cystatin-C levels also showed a positive correlation with microalbuminuria (urine albumin-creatinine ratio) in patients with type 2 diabetes mellitus.

Stroop Test Validation to Screen for Minimal Hepatic Encephalopathy in Pediatric Extrahepatic Portal Venous Obstruction.

OBJECTIVES: Minimal hepatic encephalopathy (MHE) has been reported in children with extrahepatic portal vein obstruction (EHPVO). MHE assessment is restricted to research situations as neuropsychiatric tests are time-intensive. Computerized Stroop Test (CST) has been used in cirrhotic adults for MHE screening. The study aims to assess MHE frequency in young Indian children with EHPVO and validate CST for MHE screening in pediatric EHPVO. METHODS: Thirty-seven children with EHPVO between 7 and 12 years of age and 37 age- and sex-matched controls were enrolled. Fasting plasma ammonia was measured. MHE was diagnosed by Revised Amsterdamse Kinder Intelligentie Test. The performance of a Tamil language version of CST in MHE screening was assessed. RESULTS: MHE was diagnosed in 18.9% (7/37) of EHPVO (EHPVO-MHE). Plasma ammonia levels were higher in EHPVO-MHE compared to EHPVO without MHE (EHPVO-No-MHE) but abnormal plasma ammonia levels are unsuitable for MHE screening. CST was administered in 35 EHPVO and 37 controls. EHPVO-MHE, compared to EHPVO-No-MHE, had longer "on time," "off time," "(on+off) time," and "(on-off) time." For MHE diagnosis, specificity and sensitivity of "(on+off) time" were 100% and 89.6% for a cutoff of >180.4 seconds (area under receiver operating characteristic = 0.97). CONCLUSIONS: In the absence of other risk factors for neurological insult or patent surgical shunts, MHE frequency in young Indian children with EHPVO, determined by Revised Amsterdamse Kinder Intelligentie Test, was lesser than in earlier studies. CST is suitable for MHE screening in clinical practice to select patients for neuropsychiatric evaluation.

Role of omentin 1 and IL-6 in type 2 diabetes mellitus patients with diabetic nephropathy.

AIMS: Diabetic nephropathy (DN) is one of the major chronic vascular complication of T2DM and leading cause of end-stage renal disease. Inflammation is one of the proposed pathway which explains microvascular complications in T2DM but exact mechanism is still unclear. Omentin-1 is an anti-inflammatory adipokine which promotes insulin signaling. IL-6 is a multifunctional cytokine having role in immune and inflammatory responses. The present study was conducted to elucidate the role of omentin-1 and IL-6 in the pathogenesis of DN and its association with insulin resistance. We aimed to assess and compare the serum levels of omentin-1 and IL-6 in T2DM patients with and without DN. MATERIALS & METHODS: Our study comprised of 2 groups of 41 each. Group A (controls) included T2DM without nephropathy patients and group B (cases) included T2DM nephropathy patients. Parameters studied were serum omentin-1, insulin, IL-6, fasting blood glucose, urea, creatinine, lipid profile, HOMA-IR, eGFR and BMI. RESULTS & CONCLUSION: Omentin-1 (p=0.03) was significantly decreased; concomitantly, significant increase in levels of insulin (p=0.004), IL-6 (p=0.023) and HOMA-IR (p=0.0004) were found in cases compared to controls. Bivariate analysis showed eGFR correlating positively with omentin-1 and negatively with insulin in the study population. Our study results, based on serum omentin-1 and IL-6 data suggest important role played by inflammatory mechanism and insulin resistance in the pathogenesis of diabetic nephropathy in type 2 diabetes mellitus patients.

Association Between Adiponectin and Insulin Resistance in Diabetic Urolithiasis.

OBJECTIVES: The prevalence of urolithiasis is increasing worldwide. Diabetes mellitus (DM) is characterized by insulin resistance, which increases the risk of kidney stone formation. Adiponectin is an insulin-sensitizing and anti-inflammatory cytokine, which is known to improve glucose tolerance and insulin resistance in humans. The association of insulin and adiponectin with kidney stones is not clear. Hence, the present study aim to assess the serum levels of adiponectin and insulin resistance in DM patients with urolithiasis in comparison to those without. METHODS: This study involved two groups, group A consisted of 30 patients with DM and urolithiasis, and group B consisted of 30 patients with DM but without urolithiasis (control group). Biochemical parameters studied were serum adiponectin, insulin, glucose, urea, creatinine, and 24 hours urinary calcium and phosphate. RESULTS: The serum adiponectin level was significantly increased in the diabetic urolithiasis cases (group A) compared to the control group (group B). The levels of 24 hours urine calcium and phosphorus were also significantly increased in group A. There was no significant difference in serum insulin and homeostasis model assessment of insulin resistance between the two groups. A negative correlation was seen between serum adiponectin and insulin among the cases (r = -0.368 and p = 0.045). CONCLUSIONS: We found that serum adiponectin levels are increased in patients with DM and urolithiasis.

Effect of s-methyl-L-cysteine on oxidative stress, inflammation and insulin resistance in male wistar rats fed with high fructose diet.

BACKGROUND: S-methyl cysteine (SMC) is a hydrophilic cysteine-containing compound naturally found in garlic and onion. The purpose of the present study was to investigate the protective effect of SMC on oxidative stress, inflammation and insulin resistance in an experiment of metabolic syndrome. METHODS: Male Wistar rats were divided into five groups (6 rats in each group), namely; control, control+S-methyl cysteine (SMC), high fructose diet (HFD), HFD+SMC and HFD+metformin. The 60% fructose used for 8 weeks and SMC in the dose of 100 mg/kg bw/day/rat was used in the study. The fasting glucose, insulin, insulin resistance, and tumor necrosis factor alpha and erythrocyte enzymatic antioxidants were measured. RESULTS: Increased levels of plasma glucose, insulin, malondialdehyde, tumor necrosis factor-alpha, and insulin resistance and decreased levels of glutathione, glutathione peroxidase, and catalase were found in rats on a high fructose diet. Oral administration of SMC (100 mg/kg bw/day/rat) for 60 days resulted in significant attenuation of plasma glucose, insulin, tumor necrosis factor-alpha, insulin resistance and improved antioxidant enzyme activities. CONCLUSION: Oral treatment of SMC is effective in improving insulin resistance while attenuating metabolic syndrome, inflammation, and oxidative stress in male rats fed with fructose rich diet.