Department of Biochemistry

Activities of Fetuin-A protein in type-2 diabetic nephropathy patients

2025-06-01T00:00:00Z

Activities of Fetuin-A protein in type-2 diabetic nephropathy patients Deepali, S.M 2 ABSTRACT Background: Diabetic nephropathy (DN) is a serious complication of type 1 and type 2 diabetes mellitus (T2DM). It is also known as diabetic kidney disease (DKD). It is characterized by albuminuria and decreased eGFR. Excess blood sugar leads to damage of endothelial cells, renal glomerular cells along with central and peripheral nervous system involvement.. C-reactive protein (CRP) is a systemic inflammation marker that has been revealed to be correlated with DN development. Based on albuminuria and eGFR, diabetic nephropathy is divided into 5 grades/stages by KDIGO (Kidney Disease Improving Global Outcomes) guidelines. It is a terminal disease requiring early diagnostic markers to protect renal function and individual life. Many studies have shown CRP levels increased in DN cases, showing its relation to proportional rise as the disease progress. Traditional biomarkers like serum creatinine, eGFR, albuminuria are used routinely. They have limitations in detecting early renal changes as they are influenced by age, sex, and muscle mass. Fetuin-A is an acute phase protein, inhibits receptor tyrosine kinase leading to insulin insensitivity. The Fetuin-A gene (Thr256Ser) plays a important role in Diabetes and its complications. Free fatty acids have a role in glucose intolerance resulting in diabetes which is reported by many authors. Studies have demonstrated high serum Fetuin-A levels and free fatty acids along with the elevated urinary Fetuin-A levels as early predictor of diabetic nephropathy in comparison to the usage of the traditional biomarkers like creatinine, albuminuria. Therefore, study was done to determine how Fetuin-A and its gene relate to the severity of various diabetic nephropathy stages. Aim: To estimate serum Fetuin-A levels and assess the severity of diabetic nephropathy3 Objectives: This study was under taken to estimate and compare the glycemic, renal parameters, CRP and lipid profile in controls and different stages DN cases. To estimate and compare the serum Fetuin-A, free fatty acid and urinary Fetuin-A levels in controls and various stages of diabetic nephropathy. To study the polymorphism of Fetuin-A gene in diabetic nephropathy cases. To find the best cut-off value by ROC curve analysis of these parameters for early diagnosis of diabetic nephropathy and prevent the further complications. Methods: This is a hospital based comparative study, done at medicine department, HSK hospital, in Bagalkot, Karnataka. Approval for the study was taken from institution ethical committee. Consent was taken by study participants. Confidentiality of the participants was maintained as per Helsinki declaration. 40 healthy controls and 40 type 2 diabetic nephropathy cases in each first 3 stages and 19 cases in 4th stage of diabetic nephropathy were selected between the age group of 35-65 yrs based on KDIGO guidelines for nephropathy. Serum FBS, PPBS levels were estimated by spectrophotometric method, HbA1c by HPLC, fasting insulin by CLIA, HOMA-IR by formula. Serum urea, creatinine, uric acid, urine microalbumin were estimated by spectrophotometric method, eGFR by MDRD equation. Serum TC,TGL, HDL-C, and VLDL-C were estimated by spectrophotometric method LDL-C by Friedwald formula. Serum CRP is estimated by latex turbidometric method. Serum and urinary Fetuin-A, Serum free fatty acids were estimated by ELISA method. Fetuin-A gene polymorphism study was done. Statistical analysis was done using SPSS software version 19.stages of DN progressed. Serum CRP also showed significant increase in all the stages of diabetic nephropathy compared to controls (p<0.001). Serum lipid profile showed increased levels in all the stages of DN compared to controls except the HDL-C which decreased as the stages of DN progressed (p<0.001). Serum Fetuin-A level was significantly increased in first 2 stages (p=0.003) and decreased in 3rd and 4th stages of diabetic nephropathy indicating the urinary loss of this protein. Urinary Fetuin-A was significantly increased (p=0.001) in all the stages of diabetic nephropathy. There was significant gene polymorphism noted with change in the frequency of G allele (G>C) which denotes the alteration of serum Fetuin-A levels in diabetic nephropathy cases. Serum free fatty acids also significantly increased in all the stages of diabetic nephropathy (p=0.000). Best cut-off value of serum Fetuin-A, Urinary Fetuin-A and serum free fatty acids were 48.21ng/ml, 41.28ng/ml and 395.4mmol/L respectively and area under the curve 0.802, 0.947 and 0.979. Conclusion: Serum Fetuin-A levels were significantly increased in first two stages of diabetic nephropathy but decreased in 3rd and 4th stages, whereas urinary Fetuin-A levels increased in all the stages compared to controls. There was significant positive correlation of serum and urinary Fetuin-A levels with parameters like urea, creatinine, microalbuminuria, and CRP and lipid profile observed in cases. eGFR was decreased in all the stages of diabetic nephropathy cases compared to controls, showed negative correlation with serum Fetuin-A, FFA and urinary Fetuin-A levels.Gene polymorphism showed missense mutation for the Fetuin-A gene with the 100% frequency of G allele which can be one of the factor affecting the concentration of Fetuin-A in diabetic nephropathy. Cut-off values of serum and urinary Fetuin-A, serum free fatty acids can beused in predicting the severity of diabetic nephropathy compared to other routine biomarkers of DN

Dietary inflammatory index of north Karnataka food pattern and its association with Coronary Artery Diseases

2025-06-01T00:00:00Z

Dietary inflammatory index of north Karnataka food pattern and its association with Coronary Artery Diseases Deepa, S,Sajjanar Coronary artery disease (CAD) still remains as one of the chief causes of mortality and morbidity in the world.With a pooled prevalence of 11% noted for the young adult population in India, the statistics are intimidating. The major driving force in CAD is chronic, low grade systemic inflammation which is intrinsic to the activation, progression, plaque destabilization with eventual disruption of the coronary plaques leading to Major Adverse Cardiac Events (MACEs).The risk of CAD includes an intricate reciprocity between the genetic and the factors related to an individual’s lifestyle. Of the many emeging risk factors, incessant inflammation known to be contributed by lifestyle and diet are important and are more than amenable to modification. The current study used the Dietary Inflammatory Index (DII®) to know the inflammatory potential of an individual's overall dietsry intake as it is validated by numerous studies and against many inflammatory markers. Our study was aimed to investigate the association between DII, inflammatory biomarkers, CAD severity, and MACEs in a North Karnataka population with distinct dietary patterns. Methods: This prospective cohort study was conducted over a period of two years and enrolled 310 CAD patients between the age of 18-76 year from district of Vijayapura, Karnataka, India. Dietary intake were calculated by the use of Food Frequency Questionnaire developed and validated particulary to be used in north Karnataka population. After calculating the DII scores, the CAD patients were divided into quartiles based on the DII scores. Q4 had CAD patients with higher DII scores which indicated the diet as proinflamamtory and Q1 had CAD patients with lower scores whose dietary intake were considered anti-inflammatory.Baseline demographic, anthropometric, and clinical data of the CAD patients were noted and also Serum levels of high-sensitivity C-reactive protein (hsCRP), tumor necrosis factor-alpha (TNF-α), and interleukin-10 (IL-10) were estimated as markers of chronic systemic inflammation in them. The patients were followed for 30 days to document MACEs. The software SPSS 26 (SPSS Inc., version 0.21, Chicago, IL) was employed for the statistical analyses. Our study considered p-values less than 0.05 as statistically significant.The results were analysed using statistical analyses like ANOVA, Chi-square tests, and binary conditional logistic regression, and were adjusted for the many potential confounders of CAD such as age, gender, Body MassIndex, smoking status, Hypertension, total serum cholesterol, Diabetes Mellitus(DM), physical inactivity to reduce the bias. Results: The mean DII score of the CAD cohort was 2.28 ± 1.75 which indicated a dietary pattern which is predominantly pro-inflammatory in nature. A statistically significant association was observed between higher DII quartiles (Q4) and increased prevalence of older age (p=0.01), Hypertension status (p=0.01) and status of type-2 DM (p=0.03), and serum HsCRP levels showed an increase and was statistically significant (p<0.001) and the serum levels of IL-10 levels significantly decreased (p<0.001) with increasing DII quartiles (Q1 to Q4), suggesting a direct relationship between dietary inflammatory potential (DII scores) and systemic inflammatory load. An increased incidence of MACEs (20.6% overall) was significantly associated with higher DII scores (p<0.001), particularly second MI and urgent revascularization procedures. Unadjusted analysis showed a nearly 5-fold increased risk of MACEs in the highest DII quartile (OR=4.82, 95% CI: 2.2-10.6, p<0.001). The odds of MACEs in the highest DII quartile (Q4) remained 2.52 times higher than in the lowest (Q1) even after adjusting for known confounders Conclusion: Diet with a higher DII score indicating a pro-inflammatory nature, is significantly correlated with increased systemic inflammation (higher hsCRP, lower IL-10) and a higher incidence of MACEs in CAD patients from North Karnataka. These findings underscore the crucial role inflammation associated with diet in the progression and adverse outcomes of CAD, highlighting the potential for dietary interventions as a personalized strategy for secondary prevention. Keywords: Dietary Inflammatory Index (DII), Cardiovascular Disease (CVD) Outcomes, Chronic Inflammation, Precision Nutrition, Indian Population.

Vitamin D3 Mediated Regulation of Hormone Receptors in the Pathogenesis of Triple- Negative Breast Cancer

2025-06-01T00:00:00Z

Vitamin D3 Mediated Regulation of Hormone Receptors in the Pathogenesis of Triple- Negative Breast Cancer Shankarmurthy, K N Introduction Breast cancer (BC) is the most ‘prevalent malignancy among women globally. Triple-negative breast cancer (TNBC), characterized by the absence of ER, PR, and HER2 expression, remains a particularly aggressive subtype with limited treatment options and poor prognosis.’ ‘Recent evidence suggests that the vitamin D receptor (VDR) and estrogen receptor beta 1 (ERβ1) may serve as tumor suppressors in TNBC.’ ‘This study investigates the therapeutic potential of modulating VDR and ERβ1 pathways using calcitriol and 17β-estradiol, respectively.’ Methods This study was conducted in three phases. ‘In Phase I, immunohistochemical analysis of VDR and ERβ1 expression was performed on 30 formalin-fixed, paraffin- embedded invasive ductal carcinoma samples, spanning 4 molecular BC subtypes.’ Phase II involved ‘molecular docking simulations to evaluate the binding affinities of calcitriol and 17β-estradiol to VDR, ERβ, EGFR, VEGF, and caspase-3 using Cresset Flare software.’ In Phase III, ‘in vitro assays using MDA-MB-468 TNBC cells were conducted to assess the effects of individual and combined treatments on cell viability (MTT assay) and expression of ERβ1, VDR, EGFR, VEGF, and caspase-3 (Immunoblotting).Results The results ‘of this study were obtained across three integrated experimental phases: immunohistochemistry, molecular docking, and in vitro functional assays, each contributing to a comprehensive understanding of the therapeutic relevance of VDR and ERβ1 in TNBC.’ Immunohistochemical (IHC) ‘findings from Phase I revealed that VDR and ERβ1 are variably expressed across molecular subtypes of BC. Notably, in TNBC cases, VDR was localized to both the cytoplasm and nucleus, whereas ERβ1 showed cytoplasmic expression only. This pattern was distinct from other subtypes such as Luminal A and B, which showed relatively higher nuclear staining, particularly for ERβ1. The exclusive cytoplasmic localization in TNBC suggests altered receptor signaling, possibly indicative of non-genomic pathways or receptor dysfunctionThese VDR and ERβ1 expression may play subtype-specific roles in tumor suppression and provide a basis for evaluating these receptors as therapeutic targets.’ In Phase II, ‘molecular docking simulations provided computational insights into the binding interactions between ligands (calcitriol and 17β-estradiol) and their target proteins. Calcitriol showed high binding affinity for VDR, confirming a strong ligand- receptor interaction at the predicted active site. Additionally, it exhibited moderate binding with EGFR and caspase-3, suggesting possible indirect regulatory effects on proliferative and apoptotic signaling. Similarly, 17β-estradiol demonstrated strong affinity for ERβ and VEGF, implicating its potential role in modulating estrogen- responsive and angiogenic pathways. These in silico findings support the therapeutic plausibility of targeting multiple signaling axes through ligand-mediated receptor activation.’ Phase III involved functional ‘validation through in vitro assays using MDA- MB-468 TNBC cells. Treatment with calcitriol (1–5 μM), 17β-estradiol (100–500 nM), and their combination significantly reduced cell viability in a dose- and time- dependent manner. Notably, the combination treatment produced greater reduction in viability compared to either agent alone, indicating potential additive or combination effects.’ Immunoblot ‘analysis further validated the molecular impact of treatment. Calcitriol exposure led to decreased ERβ1 expression and downregulation of EGFR and VEGF over time, while increasing caspase-3 levels, suggesting an induction of apoptosis. Treatment with 17β-estradiol similarly modulated ERβ1 expression, with limited effect on EGFR but a notable increase in VEGF modulation and caspase-3 downregulation. Most importantly, combined treatment resulted in the most robust molecular changes, showing simultaneous downregulation of proliferative and angiogenic markers and strong upregulation of caspase-3.’ Together, ‘these results demonstrate that dual modulation of VDR and ERβ1 using calcitriol and 17β-estradiol disrupts oncogenic signaling, promotes apoptosis, and may serve as an effective therapeutic strategy for ERβ1-positive TNBC.ABSTRACT Vitamin D3 Mediated Regulation of Hormone Receptors in the Pathogenesis of Triple- Negative Breast Cancer 3 Conclusion This study ‘demonstrates that dual targeting of VDR and ERβ1 using calcitriol and 17β-estradiol elicits favorable antitumor responses in TNBC cells. The combination strategy regulates key oncogenic pathways involved in proliferation, angiogenesis, and apoptosis, highlighting the therapeutic promise of receptor-based induced in ERβ1-positive TNBC. These findings warrant further validation in animal models and could contribute to the development of novel combination treatments for this aggressive BC subtype.’

Pro-angiogenic vascular endothelial growth factor (VEGF), placental growth factor (PIGF) and anti-angiogenic factor, soluble FMS like tyrosine kinase-1 (sFlt-1) in preeclampsia – A case control study

2023-05-01T00:00:00Z

Pro-angiogenic vascular endothelial growth factor (VEGF), placental growth factor (PIGF) and anti-angiogenic factor, soluble FMS like tyrosine kinase-1 (sFlt-1) in preeclampsia – A case control study Sanagappa V, Kashinkunti Background: Pregnancies that are complicated by Preeclampsia (PE) range from three to eight percent. Perinatal and neonatal mortality rate is about ten percent worldwide in developing countries due to PE. This is a multisystem disorder that leads to an increase in maternal and fetal morbidity and mortality. PE is due to impaired placentation that results in placental hypoxia, and endothelial dysfunction, It is characterized by decreased maternal circulating pro-angiogenic proteins like vascular endothelial growth factor (VEGF) and placental growth factor (PIGF), increased anti-angiogenic proteins sFlt-1, before the onset of clinical signs of PE. Oxidative stress also is implicated in the pathogenesis of PE. Studies have demonstrated low VEGF, PlGF, sFlt-1 and sFlt-1: PIGF ratio as a PE predictor but the clinical utility of these parameters is not yet generally established and only few studies have focused on neonatal outcomes. Aim: The aim of the study was to determine diagnostic criteria for diagnosis of PE using pro-angiogenic VEGF, PIGF and anti-angiogenic sFlt-1 factors. Objectives: The present study was undertaken to compare maternal VEGF, PIGF, sFlt-1 and sFlt-1:PIGF ratio in PE and normal healthy pregnant women, know the correlation between VEGF, PIGF, sFlt-1 and sFlt-1:PIGF ratio with severity of PE, to find its best cut-off value of these parameters for diagnosis of PE by ROC curve analysis and to study association with neonatal birth weight and APGAR score. Methods: This was a case control study, conducted at tertiary care hospital, in Karnataka, India. The study was approved by Institutional ethics committee. Informed consent was obtained from all the participants. Forty singleton primigravida pregnant women aged 18-35 years, 20 or more weeks of gestation, diagnosed as PE andclassified as mild/severe PE and 40 healthy pregnant women as controls were selected for the study, based on America college of obstetricians and gynecologists guidelines. VEGF, PIGF and sFlt-1 were estimated by ELISA method. All the participants were visited again to note the neonatal birth weight and APGAR score. Analysis was done using SPSS software version 19. Results: Maternal serum VEGF and PIGF were significantly (p=0.0003, p<0.0001respectively) higher in PE patients than controls and there was significant decrease in mild PE than severe PE (p=0.0465, p=0.0049 respectively) patients. Maternal serum sFlt-1 and sFlt-1:PIGF ratio were significantly higher (p<0.0001) in PE patients than normal healthy pregnant women. Best cut-off value of VEGF, PIGF, sFlt-1 and sFlt-1:PIGF ratio were 150.0 pg/ml, 105.65 pg/ml, 1843 pg/ml and 28.54 respectively , and under the curve were 0.734, 0.802, 0.945 and 0.922. Birth weight and APGAR score had significant (p=0.000) correlation with sFlt-1 and sFlt-1:PIGF ratio. Conclusion: There was a significant decrease in angiogenic factors, significant increase in anti-angiogenic factor and sFlt1:PIGF ratio in PE than healthy pregnant women. There was significant correlation between severity of PE and PIGF, sFlt1 and sFlt-1:PIGF ratio. PIGF, sFlt1 and sFlt-1:PIGF ratio with mentioned cut-off values are better markers and can be used as diagnostic markers in PE. PIGF, sFlt1 and the ratio can be used as prognostic markers with mentioned criterion values for immediate birth outcomes.