Abstracts - Basic Sciences

ROLE OF PROLONGED MILK HEATING PRACTICES AS A POTENTIAL SOURCE OF DIETARY AGES IN DIABETES *Anushka Puri, Shashikala Ranjane, Swaraj Jathar, Abhishek Dole, Monika Sharma, Dr. Mahesh J. Kulkarni CSIR- National Chemical Laboratory, Pune *E-mail: [email protected] Introduction: Milk serves as the foundational component of all dairy products. Thermal processing is essential for its nutritional preservation and storage. However, extensive literature confirms that heat treatment induces structural and chemical alterations in milk proteins. One such modification is lactosylation which diminishes protein digestibility, reduces bioavailable lysine, and generates dietary advanced glycation end-products (AGEs), thereby degrading milk’s nutritional quality. Additionally, AGEs promote oxidative stress, systemic inflammation, and insulin resistance enhancing the risk of metabolic disorders like Diabetes. Existing studies have examined temperature-dependent lactosylation, however this study investigates lactosylation via prolonged milk heating in commercial tea/coffee vending machines. With the global surge in vending machines, this poses dual risks, such as nutritional deficits from lysine loss and dietary AGE formation. Methods: Fresh buffalo milk (local dairy, Pune, India) and commercially pasteurized full-cream milk (Amul brand) were used. For prolonged heating experiments, pasteurized milk was maintained at 90 °C in a commercial tea vending machine, and samples were collected at defined time intervals. Raw and pasteurized milk samples were boiled under laboratory conditions for comparison. All total milk proteins were extracted following its fat removal by centrifugation (13,000×g, 30 min, 4 °C) and precipitation using ice-cold 10% TCA. Protein pellets were dissolved in 6 M urea in 50 mM ammonium bicarbonate, quantified by Bradford assay, and subjected to reduction (5 mM DTT) and alkylation (14 mM IAA). These proteins (100 μg) were digested with trypsin (1:40, enzyme:protein) at 37 °C, followed by formic acid quenching. Peptides were desalted using C18 cartridges, dried, reconstituted in 0.1% formic acid, and analyzed by LC-MS/MS on a TripleTOF 5600 (SCIEX) in IDA mode using a C18 reverse-phase column with a 120-minute gradient. Results: The key findings indicate a time-dependent increase in lactosylation, from 1 site to 20 sites after 2.30 hours of continuous heating. Among the six major proteins analyzed, β-lactoglobulin and α-lactalbumin were found to be more susceptible to lactosylation (43.75 % and 50% lysines modified, respectively), while caseins were less reactive (αS1-casein: 26.66%; αS2-casein: 16%; β-casein: 8.33%; κ-casein: 20%). Conclusion: This study demonstrates that continuous milk heating increases protein lactosylation, leading to accumulation of protein-bound glycation end products. Given the established role of dietary AGEs, prolonged heating practices in commercial systems may pose to be a dietary contributor to metabolic diseases such as diabetes. ROLE OF RISK LOCI OF FOXO GENES IN MODULATING THE CLINICAL COURSE AND SUSCEPTIBILITY OF TYPE 2 DIABETES IN GERIATRIC POPULATION *Indrashish Mukherjee, Narsingh Verma King George’s Medical University, Lucknow, Uttar Pradesh, India *E-mail: [email protected] Introduction: Older adults are at high risk for the development of type 2 diabetes mellitus (T2DM) due to the combined effects of genetic, lifestyle, and aging influences. The genetic contribution to lifespan is ≈25% and mainly apparent after the age of 60 years and seems to increase with age. Methods: This was an age- and sex-matched 1:1 case–control study. Two hundred and fifty-seven patients registered in the outpatient department of the Department of Medicine, KGMU, Lucknow, who were diagnosed to be the cases of T2DM were screened for the inclusion and exclusion criteria. The healthy controls were men and women above 60 years of age who had not been diagnosed with any chronic physical, congenital, or psychological illness, for which they were not on regular medication. Results: The age and gender distribution was comparable between cases and controls. Mini-Mental State Examination and homeostasis model assessment of β-cell function were significantly higher in the control group against the case group. There was no significant difference in lipid profile, fasting plasma glucose, and glycated hemoglobin between genotype groups of T2DM patients. Conclusion: This study shows that there is no association between longevity-linked rs2253310 of FOXO3 and risk of T2DM in elder North Indian population. C and G were the major and minor alleles, respectively, in both elderly T2DM patients and healthy controls. DEVELOPMENT AND EVALUATION OF A CULTURALLY TAILORED EDUCATION MODULE FOR CHILDREN WITH TYPE 1 DIABETES IN RESOURCE-CONSTRAINED SETTINGS *Latika Rohilla, Devi Dayal, Prahbhjot Malhi, Bhavneet Bharti Post Graduate Institute of Medical Education and Research, Chandigarh, India *E-mail: [email protected] Introduction: Type 1 diabetes (T1D) management among children poses unique cultural and regional challenges. This study was conducted to assess the effectiveness of a culturally tailored, education module for children with T1D on their glycemic control, health-related quality of life (HRQOL), adherence, total daily insulin dose (TDD), and frequency of hypo/hyperglycemia or diabetic ketoacidosis (DKA). Methods: Using a quasi-experimental design, 94 children (47 in each group) and their caregivers attended either the existing education or the novel module with a low reading level in Hindi/Punjabi language. The module included bedside teachings, structured group sessions, an illustrative booklet, and a low-literacy “finger chart” for estimating the correction dose for hyperglycemia. Learning evaluation was done every 3rd day, and the patient was discharged only after achieving a certain score level on the diabetes knowledge test developed and validated for this study. Clinical outcomes were assessed at baseline, 3 months, and 6 months using their diabetes logbooks. Adherence and HRQOL were assessed using the standardized Hindi versions of “Diabetes Self-Management Profile-Self Report” and “Pediatric Quality of Life Inventory 3.2 Diabetes Module.” Results: Eighty-six children completed follow-ups (mean age: 8.2 ± 4 years; 55% of females; mean duration of diagnosis: 1.9 ± 3 years). The educational level of mothers was below high school in 58%, 66% belonged to rural habitat, and 53.4% of fathers were daily wagers. Glycated hemoglobin results significantly reduced at 6 months in both the intervention group (IG) (12.2 ± 2.6–8.2 ± 1.8, P = 0.00) and the control group (CG) (12.0 ± 3–8.2 ± 1.6, P = 0.00), but between-group difference was not significant at 3 (P = 0.92) or 6 months (P = 0.73). The TDD and frequency of hypo/hyperglycemia/DKA among the groups did not vary significantly at both follow-ups. The adherence and HRQOL had a significant improvement in the IG than CG at both follow-ups (P = 0.00). Conclusion: Culturally tailored education is effective in improving the HRQOL and adherence among children with T1D living in resource-constrained settings without deteriorating their glycemic control. MITOVESICLE-ENRICHED SMALL EXTRACELLULAR NANOPARTICLES FROM BIOENGINEERED HUMANIZED INSULIN-PRODUCING NEO-ORGANS REVERSE HYPERGLYCEMIA IN A DIABETIC MOUSE MODEL *Sandeep K. Vishwakarma, Aleem A. Khan Central Research Laboratory, Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences, Hyderabad, Telangana, India *E-mail: [email protected] Introduction: Emerging evidence highlights small extracellular vesicles (sEVs) and nonvesicular extracellular nanoparticles as key mediators of intercellular communication and therapeutic cargo delivery. However, their functional diversity and translational potential remain largely unexplored. Here, we identify a novel sEV subtype – mitovesicles – secreted along with exomeres and supermeres by bioengineered humanized insulin-producing neo-organs (BHIN). These mitovesicle-enriched cargos exhibit synergistic bioactivity that restores glycemic control in diabetic mice, revealing a powerful cell-free strategy for next-generation therapeutics. Methods: BHIN were biofabricated by seeding transdifferentiated human hepatic progenitor cells into decellularized rat spleen scaffolds, recreating a physiologically relevant three-dimensional niche for β-cell function. Mitovesicles, together with exomeres and supermeres, were isolated using buoyant density-based differential ultracentrifugation and characterized by transmission electron microscopy, nanoparticle tracking analysis, western blotting, and whole transcriptome sequencing. The purified vesicles were then functionally evaluated in streptozotocin-induced diabetic mice (STZ-DM) to assess their therapeutic efficacy in restoring glycemic control. Results: BHIN cultures allow for up to 60-fold increased yields of sEVs, 40-fold of exomeres, and 65-fold of supermeres versus two-dimensional. Among the 30 isolated fractions, mitovesicles, exomeres, and supermeres were predominantly enriched in fractions F16–F20. Transcriptomic profiling revealed that mitovesicle-enriched fractions contained abundant mitochondrial mRNAs, miRNAs, and insulin-related transcripts, whereas exomeres and supermeres carried β-cell trophic miRNAs, including miR-375 and miR-21. Pathway enrichment analysis of mitovesicle fractions revealed significant upregulation of adenosine triphosphate-dependent pathways and cellular homeostasis which were not enriched in crude sEVs. Systemic administration of two consecutive doses of mitovesicle-enriched particles significantly reduced blood glucose levels, improved glucose tolerance (P < 0.05), and restored pancreatic insulin content in STZ-DM within 14 days, with effects sustained for the 60-day study period. Histological and molecular analyses demonstrated upregulation of pancreatic transcription factors and suppression of apoptotic markers, indicating sEV-mediated β-cell regeneration and functional restoration. Conclusion: By integrating advanced tissue and organ bioengineering with mitovesicle-enriched exomeres and supermeres, this approach offers an established scalable, next-generation cell-free regenerative therapeutic option for reversing hyperglycemia through functional restoration. INSUPATCH: A MADE-IN-INDIA, TUBELESS, COST-EFFECTIVE, PATCH-STYLE INSULIN INFUSION PUMP WITH NOVEL DELIVERY TECHNOLOGY *Vardhan Phadnis, Ajay Bhagwat, Ameya Pathak Renu MedTech Pvt. Ltd., Pune, Maharashtra, India *E-mail: [email protected] Introduction: Insulin pump therapy remains underutilized in India (70% adoption in developed countries), largely due to prohibitively expensive imported products, an ill-suited design for Indian context, and limited affordability of imported consumables. Prior to development, we conducted structured interviews with 100+ patients and 25+ endocrinologists/dialectologists, which highlighted consistent needs for affordability, discreet form factor, reliability, and continuous glucose monitoring (CGM) integration. This project aims to develop InsuPatch, an indigenous, tubeless, patch-style insulin infusion pump priced at approximately ₹75,000, engineered for accuracy, durability, usability, and closed-loop integration with CGM. Methods: InsuPatch employs a novel engineering mechanism that is a crucial contributor toward precision and affordability. A patent (application no. 202521092788) has been filed covering both the novel engineering and the separable, patch-style architecture. Prototyping and verification were performed at Venture Center’s ISO 13485 Class 7, 8, 9 clean-room facility. Bench testing evaluated flow-rate stability, dose accuracy, and engineering reliability. All development follows ISO 13485 design controls, risk management processes, and anticipated Central Drugs Standard Control Organization Class C regulatory pathways. Results: Early engineering tests demonstrated stable and repeatable flow rates within tolerance for and delivery. The consistent and low test thereby for insulin delivery. of the and control is with for Conclusion: InsuPatch demonstrates the of a insulin infusion pump for Indian By usability, and that have limited pump this the for pump therapy in The has potential to glycemic stability, adherence, and diabetes at a studies usability, and THE OF AND University, India *E-mail: [email protected] Introduction: is a characterized by at sites of insulin leading to insulin and glycemic control. to insulin inflammation, and both and therapeutic in diabetic patients. This study the in in and in potential of a from as a therapeutic Methods: Insulin were under conditions and with of to and were assessed using and studies were performed using insulin and to For in insulin were 14 to followed by administration at doses of and 90 reduction and were analyzed using and and Results: a significant in at in indicating of analysis revealed reduced and to extensive demonstrated a loss of at while analysis a in the structural of findings with reduced at with at revealed a of between and insulin studies a significant reduction in at all with the dose the Histological analysis reduced and Conclusion: against both in and in its therapeutic potential for the of insulin OF AND FOR AND OF DIABETIC DIABETES All India Institute of Medical Sciences, India *E-mail: [email protected] Introduction: Early of diabetic remains an in of with type 2 diabetes mellitus (T2DM) develop are – on and – to structural This study an integrating and for Methods: A study was conducted in with by into no = = and = and were quantified via and to analysis analysis of and for were in and using and were – from and to inclusion of and and were validated via was to identify Results: P = while increased with (P < both improved the the under the = = precision = = revealed a with indicating potential Conclusion: and into an model offers a and approach for The model and in A IN OF TYPE 1 DIABETES Devi Dayal, Post Graduate Institute of Medical Education and Research, Chandigarh, India *E-mail: [email protected] Introduction: of and of is the of of type 1 diabetes cells and have been as major in the regeneration of These cells are in their role in T1D remains Methods: this flow was performed on blood cells from T1D and healthy controls cells were as and the of key and including 1 6 and regulatory 4 was and were performed to Results: analysis revealed a significantly higher frequency of cells in T1D with (P < was in the to its role in A significant between and was within the (P < 0.05), a and of a of and following was in cells with cells in on and were as for therapeutic as a potential of its molecular and with and were for and validated using delivery. The of and on cells that factors may be for their and transcription factors is a therapeutic strategy to and tolerance in DEVELOPMENT OF A FOR THE OF IN IN OF DIABETES Institute of India *E-mail: [email protected] Introduction: are metabolic and are with a diabetes ketoacidosis a of diabetes for is this a a with was developed and for acid and in and Methods: The were using on a Using the were extracted into in and quantified by including and were for each Results: the demonstrated = = = within a to and a was from the indicating Conclusion: to the is and effective for analysis for of without the for prolonged in These with and OF IN TYPE 1 DIABETES Devi Dayal, Post Graduate Institute of Medical Education and Research, Chandigarh, India *E-mail: [email protected] Introduction: Type 1 diabetes (T1D) is an by of pancreatic a novel for in key are into from regulatory and in 1 to to the and pancreatic thereby Methods: performed the profiling of from T1D and 20 age- and sex-matched healthy controls and Protein with and and and as the on in T1D The for and were in the isolated from and the efficacy of was evaluated both in and in Results: the of there was a significant reduction in the of as by reduced of and as as their with reduced of and the of was significantly reduced in with with significantly the as by suppression of and the administration of in streptozotocin-induced diabetic mice to reduced and improved we to into to a studies demonstrated that their structural and in the of with the of higher to Conclusion: The of a therapeutic for evaluation of efficacy and in and A TAILORED FOR DIABETIC Indian Institute of India *E-mail: [email protected] Introduction: to the Diabetes approximately adults were living with diabetes in and this is to increase to by and by with and India the among Diabetes mellitus is a chronic metabolic characterized by hyperglycemia due to insulin or in and advanced with and for effective diabetic Methods: this a was developed by into a and The was through of of and followed by integration of to form the The was characterized for against protein using and efficacy through in and in Results: The high and for The of significantly improved against while toward its and in studies demonstrated and improved tissue regeneration to control Conclusion: The developed and its potential as an advanced therapeutic for effective diabetic OF IN Monika Sharma, Mahesh Kulkarni Chemical Laboratory, Pune, Maharashtra, India *E-mail: [email protected] Introduction: is to insulin by two at the and glycation of lysine or sites processing and leading to its potential the sites and of glycation on under conditions remain The of study was to glycation at or the and of under conditions and to the modification sites using a Methods: human was in with glucose and 50 for analysis was performed on a to the of glycation at the level and to assess the modification was carried following with trypsin and to modification with a on the and Results: analysis the of glycation on following with both in a glycation at both the and The to the consistent with increases were the of Conclusion: This study evidence that glycation at and the sites of under The of novel to the of on and their potential for insulin in diabetes. DEVELOPMENT OF A NOVEL INSULIN FOR DIABETES Jathar, Monika Sharma, Mahesh J. Kulkarni Chemical Laboratory, Pune, Maharashtra, India *E-mail: [email protected] Introduction: Insulin remains the treatment of in the management of type 1 type 2 as as diabetes. However, in proteins including The and with a group in the of insulin are to glycation by glucose and and glycated insulin has an to plasma glucose and glucose and with for advanced glycation end and insulin Additionally, pancreatic glycated insulin in A insulin that not glycation may be a for insulin therapy project Methods: this we have engineered the two using for the development of a insulin The were on an in site analysis by the were in cells and purified using One of was characterized for insulin in Results: The was characterized by and the was The glucose was done using and shows glucose to that of human treatment from the to the which that the induces glucose Conclusion: results indicate that the metabolic of human insulin while to the effects of insulin OF IN DIABETES Mahesh J. Kulkarni Chemical Laboratory, Pune, Maharashtra, India *E-mail: [email protected] Introduction: diabetes mellitus is a metabolic of that increases the risk of and Early of remains as such as fasting plasma glucose and glucose tolerance tests for glycemic and may be by These to and Protein from sustained a of molecular for risk Methods: this we evaluated the potential of glycated human as for samples were collected from healthy women = and women diagnosed with = – and – modification were quantified using Glycated in were analyzed using analysis with relevant and to assess their Results: All significantly higher in the group to the control group. analysis revealed that the demonstrated for analysis significant between and including blood and blood glucose levels, indicating its to glycemic and metabolic alterations in Conclusion: The results indicate that glycation is a of diabetes Among the analyzed, as a with potential for risk of glycated level may the of and the and management of diabetes. INSULIN AS A POTENTIAL Sharma, G Mahesh Kulkarni Chemical Laboratory, Pune, Maharashtra, India *E-mail: [email protected] Introduction: the insulin Glycated insulin is to insulin but has an function. the of insulin toward its and its which to a more insulin resistance to insulin insulin to glucose which on leading to the insulin resistance the increased insulin is to the and the hyperglycemia and oxidative in to the and of glycated insulin in the Methods: plasma samples were collected from each of and diabetic from Diabetes and were collected for all the developed to identify and insulin and glycated insulin by monitoring using we quantified the level of insulin by and did of insulin by and and the effects of high glucose on insulin in Results: The level of insulin is more to insulin as is more reactive and the glucose and treatment the insulin from Conclusion: The developed between and insulin in This be to the of functional insulin and be in a treatment for the OF IN Mahesh Kulkarni Chemical Laboratory, Pune, Maharashtra, India *E-mail: [email protected] Introduction: is a in and and is for its and abundant such as and glucose and which in glycation with groups in This to the of (AGEs), which are in and chronic metabolic AGE and tissue Methods: samples were subjected to both and were extracted using a followed by analysis using a C18 column under and profiling of while was for of glycated were at 30 and at a of with glycated using a 1:1 0.1% formic Results: analysis with in mode and in analysis revealed glycated including and of and lysine, as as glycated acid, and high such as glycated and glycated indicating glycation within the Conclusion: This study demonstrates that a of glycated extensive between abundant and The of glycation that may as a dietary of AGEs, which are to diabetic and metabolic These findings the of glycation in and the for to identify glycated proteins in and to glycation