Safia Habib

Introduction: Type 2 Diabetes Mellitus (T2DM) involves chronic inflammation and recurrent hyperglycemia, often linked to excessive fructose intake. Some patients develop autoantibodies, indicating immune dysfunction. Fructose promotes protein glycation and the formation of Advanced Glycation End-products (AGEs), which may expose novel epitopes recognized as foreign. This study examined the antigenicity of fructosylated human insulin and its impact on lymphocytes in diabetic patients. Methods: Fructosylated insulin was characterized using spectroscopic techniques, with aggregation assessed by ThT, ANS, and DLS, and conformational changes analyzed via CD spectroscopy. Biochemical and biophysical methods confirmed the presence of structural alterations. Antigenicity was evaluated using ELISAs in sera from 40 healthy controls and 40 newly diagnosed T2DM patients, with ROC and AUC analyses determining prognostic relevance. Results: Concentration-dependent increase in AGE-specific fluorescence, increased hydrophobicity, and significant loss of lysine (48.6%) and arginine (58.8%) residues, accompanied by conformational changes. Direct binding ELISA revealed statistically significant reactivity (p ≤ 0.05) with diabetic sera. Competition ELISA demonstrated 55% ± 3.7% inhibition for fructosylated insulin compared to 31.4 ± 5.1% for native insulin. IgGs isolated from diabetic sera exhibited 96.67% specificity for fructosylated insulin, with sensitivity and specificity analyses yielding AUC = 1.000 (p < 0.01). Discussion: Fructosylation causes changes in insulin chemistry, which leads to altered structural, conformational, and antigenic potential in T2DM patients. Conclusion: Screening for anti-fructosylated insulin antibodies may help identify T2DM patients with hyperglycemia-associated, subclinical autoimmune activity linked to abnormal insulin responses.

Background: Doctoral training involves prolonged performance demands, uncertainty, supervisory dependence, and career pressure, yet objective biochemical correlates of academic stress remain incompletely characterized. This study examined academic stress in relation to glycemic, neuroendocrine, inflammatory, and thyroid-related markers, alongside psychological symptoms and sleep quality, in doctoral research scholars. Methods: This cross-sectional study included 203 doctoral research scholars (123 female, 80 male; mean age 27.2 years). Academic stress was assessed using a 34-item doctoral-adapted Academic Stress Scale (AST). Glycated hemoglobin (HbA1c), evening cortisol, C-reactive protein (CRP), and thyroid-stimulating hormone (TSH) were measured as biochemical outcomes. Depression, anxiety, and stress symptoms were assessed using the 21-item Depression Anxiety Stress Scales (DASS-21), and sleep quality using the Pittsburgh Sleep Quality Index (PSQI). Primary associations were examined using linear regression with heteroscedasticity-consistent type 3 (HC3) robust standard errors, adjusted for age, gender, body mass index, and lifestyle activity, with false-discovery-rate (FDR) correction. Results: Mean AST was 109.5 (standard deviation 21.0). Above-normal DASS-21 scores were observed for depression in 64.5%, anxiety in 86.7%, and stress in 49.3%, while 79.3% had poor sleep quality. Higher AST was associated with all four biochemical markers. Per 10-point increase in AST, HbA1c was 0.225 percentage points higher (95% confidence interval 0.158–0.291), evening cortisol 0.502 µg/dL higher (0.299–0.706), CRP 13.9% higher (8.4–19.7%), and TSH 0.125 µIU/mL higher (0.049–0.200); all FDR q ≤ 0.0013. Depression and anxiety severity distributions differed by gender, whereas stress did not. Exploratory analyses showed positive correlations between DASS-21 symptoms and all four biomarkers (Spearman ρ = 0.22–0.58; all FDR q < 0.01). Conclusions: Higher academic stress was associated with variation across glycemic, neuroendocrine, inflammatory, and thyroid-related markers in doctoral scholars, alongside substantial psychological symptom and sleep burden. The cross-sectional design precludes conclusions regarding temporal direction or causality and supports the need for longitudinal research on doctoral health and well-being.

Chronic stress is a major public health concern, contributing to a spectrum of physiological and psychological disorders, including anxiety, depression, cardiovascular diseases, and metabolic syndromes. This review examines the epigenetic mechanisms mediating the impact of chronic stress on human health, focusing on DNA methylation, histone modifications, and non-coding RNAs. These epigenetic alterations act as dynamic interfaces between environmental stressors and genomic responses, modifying gene expression in critical stress-related pathways, such as the hypothalamic- pituitary-adrenal (HPA) axis and neurotrophic factors like BDNF. The significance of this research lies in elucidating how these epigenetic changes, induced by early-life adversity, socioeconomic pressures, and environmental toxins, create lasting molecular imprints that increase susceptibility to stress-related disorders and can be transmitted across generations. Key findings include stress-induced hypermethylation of the glucocorticoid receptor gene (NR3C1), histone acetylation changes in brain regions like the hippocampus and amygdala, and the regulatory roles of microRNAs (e.g., miR-132) and long non-coding RNAs (e.g., HOTAIR). These modifications disrupt neural plasticity, emotional regulation, and cognitive function, while also contributing to physical health outcomes, including inflammation, metabolic dysregulation, and cancer. The review also explores therapeutic interventions, such as DNA methylation inhibitors (e.g., 5-azacytidine), histone deacetylase inhibitors (HDACIs), and lifestyle modifications like mindfulness and dietary polyphenols. By integrating molecular insights with clinical perspectives, this article underscores the critical role of epigenetic research in developing targeted interventions to mitigate the enduring effects of chronic stress, offering transformative potential for personalized medicine and public health strategies.

Introduction: Fructose, like other sugars and sugar metabolites, is capable of glycating protein. Insulin's fructosylation leads to the generation of Advanced Glycation End Products (AGEs). Reducing sugars reaction with proteins to form Schiff’s bases, which are characterized by the presence of an imine (C=N) bond. The Schiff bases then undergo irreversible rearrangements, followed by the production of much more stable compounds called Amadori products. These Amadori products can further undergo oxidation, dehydration, cyclization, and condensation to form highly toxic advanced glycation end-products (AGEs). These processes are accompanied by oxidative stress, secondary structural perturbations, and altered morphology, progressing toward amyloidogenesis. Metformin, a biguanide, is the most common drug used to treat type 2 Diabetes Mellitus (T2DM). Aim: The aim of this study was to evaluate the protective effect of metformin against fructosylation-induced cross-β structures and amyloid aggregations of human insulin. Methods: UV-absorbance and fluorescence spectroscopy, determination of carbonyl content, free lysine and arginine residues, determination of fructosamine content, SDS-PAGE, circular dichroism (CD) spectroscopy, dynamic light scattering, and scanning and transmission electron microscopy. Results: Physicochemical studies in the presence or absence of metformin revealed a concentration-dependent structural restoration of fructosylated insulin. Results from the thioflavin-T fluorescence assay suggested that metformin limited the transition of insulin from native to fibrillar state, which was validated by scanning and transmission electron microscopy. Metformin lowered the ThT fluorescence intensity in a concentration- dependent manner. The ThT-specific fluorescence intensity was reduced to 114 and 112.5%. The fluorescence intensity at 2.5 mM metformin was close to native insulin. Electron microscopy revealed that insulin fructosylated by 25 mM fructose in the presence of 2.5 mM metformin suppressed the formation of fibrillar structures. Dynamic light scattering data revealed the potential of metformin to conserve and reinstate the increased hydrodynamic radii (Rh) of fructosylated insulin close to the native conformer. The Rh values of native, fructosylated insulin and insulin incubated with fructose and metformin were found to be 2.65 ± 0.28, 307.6 ± 24.19 nm, and 110.1 ± 4.08 nm, respectively. This study also identified metformin as an antioxidant by protecting critical amino acid residues of the insulin domain. Discussion: The study reports the protective effects of metformin on insulin structure, conformation, and function. The findings suggest a potential role for metformin in improving the risk profile associated with insulin resistance due to altered structure or the accumulation of protein aggregates. Interaction studies between insulin and metformin presented here are due to the chemical effect; hence, further in-depth studies are required to identify the molecular mechanism of insulin sensitivity and changes in cellular processes and pathways. Conclusion: The results suggest that metformin safeguards against fructosylation-induced structural, conformational, morphological, and amyloidogenic aggregating tendencies of insulin. Protein aggregation has been linked to several neurological and metabolic diseases. Hence, metformin may be crucial in preserving the biological activity of insulin by maintaining and protecting its structural integrity and minimizing the associated comorbidities. The study may further be extended to identify the role of metformin in controlling the gradual insulin resistance in T2DM at the molecular level.

Ovarian cancer (OC) ranks as the fifth leading cause of cancer-related deaths in the United States, posing a significant threat to female health. Late-stage diagnoses, driven by elusive symptoms often masquerading as gastrointestinal issues, contribute to a concerning 70% of cases being identified in advanced stages. While early-stage OC brags a 90% cure rate, progression involving pelvic organs or extending beyond the peritoneal cavity drastically diminishes it. Overcoming chemoresistance and metastasis requires a deep understanding of the associated progression mechanisms for innovative therapies. Extracellular vesicles (EVs), containing proteins, RNAs, DNAs, and metabolites, have surged in recent years, significantly impacting tumor progression, recurrence, immune evasion, and metastasis associated with the ovarian tumor microenvironment. Recent research unveils organ-specific metastatic patterns in OC, providing insights into tumor cell interactions and signaling crosstalk with stromal cells. The review explores the role of EVs behind OC cell metastasis and chemoresistance. Furthermore, the article delves into the role of EVs in the tumor microenvironment, immune evasion, and as biomarkers in context to OC, offering promising therapeutic strategies to enhance survival rates for OC patients. Lastly, the article focuses on an overview of PI3K/AKT/mTOR, MAPK/ERK, and VEGFR signaling pathways in the pathophysiology of ovarian cancer.