Doctoral Theses

Wnt/β-catenin signaling in hypoxia-induced pulmonary artery smooth muscle cell proliferation - Role of bioactive molecule of Mucuna pruriens

Sun, 01 Jun 2025 00:00:00 GMT

Wnt/β-catenin signaling in hypoxia-induced pulmonary artery smooth muscle cell proliferation - Role of bioactive molecule of Mucuna pruriens Supriya, Bhosale Introduction: Pulmonary hypertension (PH) is a progressive, life-threatening disease characterized by vascular remodeling, constriction, and thrombosis, primarily driven by excessive proliferation of pulmonary arterial smooth muscle cells (PASMCs). Hypoxia is a key trigger in PH pathogenesis. The Wnt/β-catenin signaling pathway, critical for cell fate, migration, and organogenesis, plays a pivotal role in PH, with β- catenin mediating transcriptional activation of target genes upon Wnt ligand stimulation. Targeting Wnt/β-catenin signaling represents a promising therapeutic strategy for PH. This study examines its role in hypoxia-exposed PASMCs and evaluates the therapeutic potential of gallic acid and β-sitosterol through in-silico and in-vitro approaches. Objective: To study the role of isolated biomolecules from Mucuna pruriens gallic acid and β-sitosterol on Wnt/ β-catenin mRNA expression in the human pulmonary artery smooth muscle cells exposed to hypoxia. Method: The current study used a computational method based on the ligand-protein interaction technique to determine the therapeutic potential of gallic acid and β- sitosterol with the Wnt/ β catenin pathway. The same compounds are used to investigate. The Invitro study explored the role of gallic acid and β-sitosterol in hypoxia-exposed PASMC lines. Result and Discussion: The current study identified different pharmacological properties of gallic acid and β-sitosterol bioactive molecules to analyze the in silicoADME/T properties. All were within Lipinski’s rule acceptable range, and molecular docking analysis showed that β-sitosterol has more interaction sites with Wnt5a. The Invitro study revealed that when HPASMC is exposed to hypoxia, there is downregulation of the Wnt5a gene and upregulation of the β-catenin gene. β-sitosterol and gallic acid can be attributed to inhibiting the β-catenin pathway via the downregulation of β-catenin gene expression. Conclusion: The present study focused on in-silico phytochemical analysis and in vitro investigations to evaluate the potential therapeutic role of isolated biomolecules from Mucuna pruriens seed extract β-sitosterol and gallic acid in hypoxia-exposed pulmonary artery smooth muscle cells (HPASMCs). These findings suggest that Mucuna pruriens, or its bioactive molecule gallic acid and β-sitosterol, may exert protective effects against hypoxia-induced vascular remodeling by targeting the Wnt/β-catenin signaling pathway.

Modulation of NFKB ligand RANKL Signaling Lipopolysaccharide Induced Rheumatoid Arthritis Cell Line by Pithecellobium dulce.

Sun, 01 Jun 2025 00:00:00 GMT

Modulation of NFKB ligand RANKL Signaling Lipopolysaccharide Induced Rheumatoid Arthritis Cell Line by Pithecellobium dulce. Soumya T, Tungal ntroduction: In the world, 1% of people suffer from chronic inflammatory autoimmune diseases like rheumatoid arthritis (RA). The individual with rheumatoid arthritis suffers from synovial inflammation, cartilage degradation, joint destruction, leading to reduced quality of life. The disease pathogenesis involves the activation of pro- inflammatory cytokines and NF-κB ligand RANKL signaling pathway. The research on plant-based drug design and discovery is to target pathways to find promising phytochemical alternatives for relieving symptoms of RA and other chronic diseases. This study aimed to investigate the role of bioactive compounds Pithecellobium dulce gallic acid and quercetin, in modulating the NF-κB ligand RANKL signaling pathway by targeting MMP-1 expression through in silico and in vitro approaches. Objective: To evaluate the effect of isolated bioactive compounds from Pithecellobium dulce fruit gallic acid, and quercetin on mRNA expression of MMP-1 in NF-kB ligand RANKL signaling. Method: The study involved In Silico and experimental in vitro methods. Ligand– protein interaction studies were conducted to assess gallic acids and quercetin’s therapeutic potential in targeting the NF-κB ligand RANKL signaling pathway. Additionally, green-synthesized Pd-Cu nanoparticles (Pd-CuNPs) were characterized and evaluated. Pharmacokinetic properties, drug-likeness, gastrointestinal absorption, blood-brain barrier permeability, and solubility were predicted using ADME/T tools. Molecular interaction and molecular dynamics simulation were performed to identify potential interactions. The gene expression of MMP-1 was evaluated through RT-PCR in treated rheumatoid arthritis cells. Results: In silico analysis confirmed gallic acid and quercetin have favourable pharmacokinetic and safety profiles. Docking and simulation studies identified strong binding affinities with target proteins. Experimental In vitro results revealed a significant downregulation of MMP-1 mRNA expression in treated RA cells, gallic acid (0.299 ± 0.25, p < 0.05), Pd-CuNPs (0.432 ± 0.22), quercetin (0.519 ± 0.01), and crude extract (0.633 ± 0.03), compared to control. Conclusion: The study demonstrated that gallic acid, quercetin, and Pd-CuNPs exhibit promising modulatory effects on the NF-κB ligand RANKL signaling pathway. These findings support their potential as plant-based alternative agents for managing inflammatory disease like rheumatoid arthritis

Development of Skill-based Competency Module of Community Medicine subject by using Community Based Medical Education approach for Indian Medical Graduates

Sun, 01 Jun 2025 00:00:00 GMT

Development of Skill-based Competency Module of Community Medicine subject by using Community Based Medical Education approach for Indian Medical Graduates Praveen, Gangnahalli Introduction: The undergraduate medical education programme is designed with a goal to create an “Indian Medical Graduate” (IMG) possessing requisite knowledge, skills, attitudes, values and responsiveness, so that she or he may function appropriately and effectively as a physician of first contact of the community while being globally relevant. Thus, it is an approach in which the focus of teaching–learning and assessment is on real-life medical practice. Objectives: To Develop, Validate & Assess the Impact of skill-based Competency Module of Community Medicine subject by using Community-based medical education approach. Methodology: This study aims to Design, validate & implement skill-based competency module for and to take feedback from medical undergraduate students. Materials & Methods: An observational study was conducted by preparing the draft of the selected skill-based competencies of the community medicine subject named as community diagnosis module and validated with help of experts of subject and health education profession by using content validity Index. This is followed by the implementation of the module to second MBBS students, assessment of the outcome and feedback collection. Results: The Content Validity Index, based on assessments from ten field experts, yielded a score of 0.86, indicating good validity. Feedback analysis from medical student’s post-implementation revealed that transitioning from classroom teaching to community experiences was the most valued aspect of the program. Student feedback indicated a favourable consensus on various aspects of the module, including its objectives, teaching methods, materials, assessments, and active participation in activities. Conclusion: The development of a skill-based Competency Module for Community Medicine, by using community-based medical education approach, holds significant promise for improving the competency and skills of Indian Medical Graduates. This module will equip them with the knowledge and skills necessary to address the complex and evolving public health challenges faced by communities

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

Sun, 01 Jun 2025 00:00:00 GMT

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