Gestational Weight Gain (GWG) affects maternal and fetal health. Vitamin D insufficiency, common during pregnancy, may impact weight control via metabolic, inflammatory, and endocrine pathways. However, the association between maternal vitamin D and GWG remain unclear. Therefore, this narrative review's objective is to integrate existing evidence on the correlations between maternal vitamin D and GWG, examine the effect of pre-pregnancy Body Mass Index (BMI), assess the cumulative effects of vitamin D deficiency and inadequate GWG on pregnancy outcomes, and assess the efficacy of vitamin D supplementation. A narrative review was conducted in accordance with SANRA 2.0 principles, with the search strategy reported in line with PRISMA-S guidance. A literature review was conducted in PubMed, Scopus, and Google Scholar to identify studies examining gestational weight gain (GWG) and serum 25-hydroxyvitamin D status published between January 2015 and November 2025. Observational and interventional studies were eligible for inclusion. After eliminating duplicates, reviewers independently evaluated 1,395 records; 14 studies were included. The prevalence of vitamin D deficiency and its bidirectional associations with maternal weight, gestational weight gain as a modifier of vitamin D's metabolic effects, combined effects on fetal and offspring outcomes, and supplementation trial evidence were narratively synthesized. Vitamin D insufficiency affected 30-85% of pregnant women worldwide. Bidirectional vitamin D-GWG relationships differed among studies, with prepregnancy BMI as a substantial effect modifier. In overweight women, low first trimester vitamin D levels were associated with 3.70 kg higher total GWG. Vitamin D's insulin resistance protection was greatly reduced by excessive GWG. High-dose supplementation (1600 IU/day) met biochemical needs but did not affect GWG. In one cohort, both low vitamin D and suboptimal GWG independently predicted small-for-gestational-age births. The association between maternal vitamin D status and gestational weight gain remains inconsistent across the current evidence base. Pre-pregnancy BMI may modify this relationship, with the strongest associations observed among overweight and obese women. Integrated strategies addressing both vitamin D adequacy and appropriate gestational weight gain, particularly among women with elevated pre-pregnancy BMI, should be explored in future research. Vitamin D supports skeletal health and also contributes to immune and metabolic function. In pregnancy, achieving recommended gestational weight gain is important because both inadequate and excessive gain are linked to adverse outcomes for mothers and infants. This narrative review summarises global evidence on whether maternal vitamin D status is associated with gestational weight gain and whether pre-pregnancy body weight influences this relationship. We searched major bibliographic databases for studies published between 2015 and 2025 that measured vitamin D during pregnancy and reported gestational weight gain. Across studies, results were inconsistent. Some reported associations between lower vitamin D status and higher or lower gestational weight gain, while others found no clear relationship. Methodological differences likely explain part of this variability, including timing of vitamin D assessment, definitions of deficiency, measurement of weight gain, and adjustment for key factors such as pre-pregnancy body mass index, season, diet, physical activity, and supplementation. Although evidence remains mixed, higher pre-pregnancy adiposity is frequently associated with lower circulating vitamin D and may also relate to a greater likelihood of non-recommended gestational weight gain and metabolic complications. These overlapping risks highlight the importance of considering baseline maternal weight status when interpreting the vitamin D and gestational weight gain literature. Overall, current evidence does not establish a consistent causal link between vitamin D status and gestational weight gain. Robust prospective studies and adequately powered trials are needed to clarify directionality and determine whether optimising vitamin D status, within comprehensive antenatal care, supports healthier gestational weight trajectories.
Use of Medicare Advantage (MA) insurance has been associated with lower cancer treatment costs than traditional Medicare (TM). Whether there are differences in guideline-concordant care or treatment costs for beneficiaries insured under MA vs TM is unknown. To assess whether Medicare insurance type is associated with receipt of optimal cancer treatment and costs. Retrospective cohort study of a population-based sample using Medicare claims and encounter data from 2015 through 2019. Participants were Medicare beneficiaries with continuous enrollment in either MA or TM and an incident cancer diagnosis from 2016 through 2019. Included patients were those with cancer treatment scenarios (eg, metastatic melanoma, adjuvant therapy for stage III colon cancer) that had substantial variation in the costs of recommended treatment options. Data were analyzed from April 2025 to March 2026. Medicare insurance type of MA or TM. The pharmacologic cancer treatment among the available options each patient initiated was analyzed using Medicare data. Each patient's treatment was linked by diagnosis date to contemporary National Comprehensive Cancer Network Guidelines recommendations and Medicare reimbursement rates to ascertain the coprimary outcomes whether a patient received the optimal treatment for their cancer type and the anticipated cost of the initiated treatment. The association of outcomes with insurance type was accessed using generalized estimating equations to estimate risk ratios (RRs) after balancing patient characteristics through inverse probability-of-treatment weighting. Models were clustered at the physician level and adjusted for scenario, diagnosis year, scenario × diagnosis year interaction, and oncologist characteristics. Of 35 245 patients (median age, 74 [IQR 70-79] years; 63.2% male), 24 269 had TM and 10 976 had MA. The median time to treatment initiation was 36 (IQR, 20-60) days for MA vs 35 (IQR, 19-59) days for TM. The unadjusted mean (SD) treatment cost was $29 252 ($80 391) for MA vs $40 874 ($106 205) for TM. MA beneficiaries had a likelihood of optimal treatment similar to TM beneficiaries (adjusted RR, 0.99; 95% CI, 0.97-1.02), and MA was associated with lower treatment cost (adjusted cost ratio, 0.94; 95% CI, 0.91- 0.97). Mean savings within this patient cohort was -$931 (95% CI, -$1244 to -$615). In this cohort study of Medicare beneficiaries with cancer, MA beneficiaries had lower estimated treatment costs and similar likelihood of receiving optimal treatment compared with TM beneficiaries.
Biological ageing is a heterogeneous process that shapes susceptibility to chronic disease. However, whether ageing patterns diverge within clinically defined type 2 diabetes (T2D) subgroups remains unclear. We investigated whether epigenetic age acceleration (EAA) differs across T2D phenotypes and whether these patterns relate to subsequent renal vulnerability. We included 607 multi-ethnic Asians with recent-onset T2D previously classified into three clinically distinct subgroups: mild obesity-related diabetes (MOD), mild age-related diabetes with insulin insufficiency (MARD-II), and severe insulin-resistant diabetes with relative insulin insufficiency (SIRD-RII). EAA was quantified using multiple epigenetic clocks. Kidney function was assessed longitudinally using eGFR slope over a median follow-up of 7.3 years, serving as a maker of cumulative systemic ageing burden. SIRD-RII, despite being chronologically younger, showed accelerated ageing across multiple second-generation clocks, whereas MARD-II, despite being chronologically older, showed comparatively lower levels of EAA. Among all epigenic clocks examined, GrimAge2logA1c was both highest in SIRD-RII and the only clock associated with kidney function decline (eGFR slope, β [SE] = -0.113 [0.040], p = 4.77 × 10-3). Epigenome-wide analyses identified subgroup-specific, glucose-responsive loci associated with accelerated ageing. Hypomethylation at TXNIP (cg19693031) characterised the SIRD-RII subgroup and was associated with both faster ageing pace and kidney function decline. Exploratory longitudinal within-person changes in glucose-linked epigenetic ageing were correlated with changes in TXNIP methylation within SIRD-RII (r = -0.901, p = 1.93 × 10-6). These findings suggest that T2D subgroups represent distinct epigenetic ageing phenotypes that may be associated with renal vulnerability. Integrating epigenetic ageing metrics with clinical stratification may enhance identification of individuals at risk for accelerated systemic ageing, informing geroscience-guided intervention strategies early in disease development.
The Delphi method is widely used to derive expert consensus on complex clinical problems, yet it is slow and resource intensive. Recent advances in large language models and retrieval‑augmented generation (RAG) offer the possibility of accelerating consensus while maintaining methodological rigor. Large language models can retrieve and summarize evidence, but they frequently hallucinate and cannot reliably cite sources. At the same time, RNA‑based drugs and messenger RNA vaccines are rapidly moving from concept to clinic, generating a pressing need for timely, evidence‑based consensus on regulatory, manufacturing, and clinical issues. We evaluated whether a modular, RAG‑enabled, multi‑agent artificial intelligence (AI) pipeline could replicate the post-round 1 behavior of a human reviewer in a Delphi study. The primary objective was to determine whether AI‑assisted statement revision could rescue a greater proportion of subthreshold statements and achieve a consensus comparable to that obtained through human revision by round 2. A parallel, 2‑arm Delphi study was conducted on 28 statements about RNA medicines. In total, 50 international panelists (clinicians, researchers, and patient representatives) were randomized into human (arm A) and AI‑assisted (arm B) groups. After round 1, statements below the 75% agreement threshold were revised either manually by human reviewers or by an AI pipeline comprising the following software agents: (1) ReferenceDetector, to identify external citations; (2) Summarizer, to produce structured summaries of supporting PDFs; (3) a hybrid RAG module that combined dense and sparse retrieval with cross‑encoder reranking; and (4) Refiner, which generated revised statements, reasoning logs, and explicit citations. Two human reviewers with expertise in Delphi methodology and literature review who had contributed to statement development verified retrieved citations and approved or amended revisions. Agreement rates and vote distributions were compared across arms. Arm A reached consensus on 71.4% (20/28) of the statements in round 1, whereas arm B reached consensus on 46.4% (13/28). After revision, consensus increased to 92.9% (26/28) of the statements in arm A and 85.7% (24/28) in arm B. The AI arm exhibited a larger mean improvement (absolute difference between rounds 1 and 2=39.3 percentage points) because more statements were initially below the threshold. Nonetheless, the absolute difference between arms after round 2 was modest (7.2 percentage points). AI‑assisted revisions were particularly effective for statements far below the threshold, but both arms failed to rescue 2 to 3 statements owing to substantive disagreements. A modular, citation‑anchored AI pipeline can closely approximate human performance in Delphi consensus procedures while substantially reducing manual workload. When paired with human oversight, AI assistance accelerated revision and closed most of the performance gap by the second round. Adoption of AI‑assisted workflows could accelerate consensus development on emerging technologies such as RNA therapeutics provided that transparency, rigorous retrieval, and human review are maintained.
Colony-stimulating factor 1 receptor (CSF1R) inhibitors, such as PLX5622 and PLX3397 (pexidartinib), are widely used for in vivo microglial depletion and for investigating microglial functions and therapeutic potential. Although CSF1R inhibitor-based studies have uncovered important roles for microglia in processes, such as anesthesia, addiction, and obesity, whether the resulting phenotypes reflect microglial depletion alone remains increasingly debated. Our previous work has shown that PLX5622 activates hepatic constitutive androstane receptor (CAR)-dependent xenobiotic metabolism, altering the metabolism of anesthetics and addictive drugs, and amplifying apparent microglial phenotypes. Whether other CSF1R inhibitors, particularly the FDA-approved PLX3397, exert systemic metabolic effects that may influence the interpretation of brain phenotypes remains unknown. Here, we demonstrate that PLX3397 exerts hepatic metabolic effects that are mechanistically distinct from those induced by PLX5622. Although PLX3397 only weakly affects xenobiotic metabolism, it markedly enhances endogenous hepatic lipid metabolism, inducing a fasting-like state characterized by increased lipid utilization and ketogenesis despite the absence of nutrient deprivation. By uncovering previously unrecognized peripheral effects of PLX3397, our findings identify brain-periphery interactions as a potential source of confounding in studies of microglial function. These results suggest that systemic metabolic effects should be carefully considered when interpreting neural or behavioral phenotypes in pharmacological microglia depletion paradigms.
Suboptimal transitions from pediatric to adult health care can negatively impact disease outcomes, medication regimen adherence, and disease self-management in adulthood. Little is known about whether the presence of coping strategies in young adults (YAs) with sickle cell disease (SCD) is associated with higher transition readiness scores. To evaluate whether coping strategies and social support are associated with higher transition readiness scores in YAs with SCD. This cross-sectional study was a subanalysis of baseline data from a prospective randomized clinical trial of YAs aged 17 to 25 years with SCD planning to transition to an adult sickle cell clinic within the next 12 months at 5 institutions across Connecticut, New York, Ohio, and Pennsylvania. The recruitment period was from January 15, 2019, to December 31, 2022. Analysis used data from this time period and was conducted from September 30, 2024, to June 30, 2025. Measures included the Transition Readiness Assessment Questionnaire (TRAQ) to assess transition readiness, the Medical Outcomes Study-Social Support Survey (MOS-SSS) emotional/informational subscale to assess social support, and the Brief-COPE (problem-focused, emotion-focused, and avoidant subscales) to assess coping strategies. Bivariate and multivariable linear regression analyses were used to assess differences of transition readiness. Covariates included worry (PedsQL Sickle Cell Disease Module worry I), age, gender, SCD disease severity, and social support. The final cohort included 373 YAs (mean [SD] age, 18.9 [1.9] years; 190 female [51.5%]), with 335 of 367 reporting race as Black (91.3%) and 335 of 365 reporting ethnicity as non-Hispanic/Latino (91.8%). Unadjusted analysis found that overall coping (mean difference, 0.14; 95% CI, 0.01-0.27; P = .03), problem-focused coping (mean difference, 0.18; 95% CI, 0.09-0.27; P < .001), and the emotional/informational subscale of the MOS-SSS (mean difference, 0.13; 95% CI, 0.07-0.19; P < .001) were associated with higher transition readiness scores. Adjusted linear regression analysis showed that problem-focused coping (mean difference, 0.10; 95% CI, 0.003-0.19; P = .04) was associated with transition readiness even after adjusting for age, gender, disease severity, worry, recruitment site, and social support. In this cross-sectional study in a national cohort of adolescents and YAs with SCD, problem-focused coping strategies were associated with higher levels of transition readiness. These findings suggest important intervention targets for supporting transitioning adolescents with SCD.
Autophagy was originally identified as a survival mechanism to allow cells to survive under nutrient-deprived or stressful conditions whereas cellular senescence was considered a tumor-suppressive mechanism. Both processes can be induced by similar stimuli and can influence each other. There have been continued debates about whether they are causally linked, whether autophagy promotes or prevents senescence or if they are independent of each other. Protein kinases play integral roles in cell fate decision and have a major influence on both autophagy and senescence. While mechanistic target of rapamycin complex 1 is considered the master regulator of autophagy, it also influences senescence. Mitogen-activated protein kinases originally associated with senescence can regulate autophagy. While there have been numerous review articles on the interplay between autophagy and senescence, a comprehensive review on how various kinases participate in this interplay is lacking. The purpose of this review is to learn lessons from some old and recent studies to understand how kinases contribute to this changing field. Since both autophagy and senescence can have beneficial and detrimental effects and kinases are important drug targets, insights regarding how kinases orchestrate these two processes should help develop therapeutic strategies to treat diseases, such as aging and cancer.
Rare cases of myocarditis and pericarditis following mRNA coronavirus disease-19 (COVID-19) vaccination have been reported, primarily among males under 30 years of age. Although the underlying mechanisms remain unclear, cardiovascular risk factors such as hypertension, diabetes, dyslipidemia, and obesity have been hypothesized as potential contributors. Using the Vaccine Adverse Event Reporting System (VAERS), this study investigated whether these comorbidities were disproportionately reported among young adult males with myocarditis or pericarditis following mRNA COVID-19 vaccination compared with vaccinated and general populations. Additionally, this study represents a novel application of disproportionality principles in spontaneous reporting databases to explore, beyond signal detection, the factors that might be associated with the occurrence of an adverse event. An observed-versus-expected analysis was conducted using reports of myocarditis or pericarditis following mRNA COVID-19 vaccination among males aged 18-30 recorded in VAERS between 2021 and 2022. The prevalence of hypertension, diabetes, dyslipidemia, and obesity among reported cases was compared with prevalence estimates derived from epidemiological studies of vaccinated populations and national population statistics. In addition, disproportionality analyses were performed using comparator groups consisting of reports associated with other vaccines and other adverse events following immunization (AEFIs) within VAERS. A total of 859 eligible reports of myocarditis/pericarditis following mRNA COVID-19 vaccination were identified among males aged 18-30. The prevalence of hypertension, diabetes, dyslipidemia, and obesity was 0.35%, 0.58%, 0.81%, and 1.75%, respectively and was significantly lower than those reported in vaccinated and prepandemic general populations. In disproportionality analyses, myocarditis/pericarditis reports following mRNA vaccination showed higher prevalence of dyslipidemia and obesity, but lower prevalence of hypertension, compared with selected VAERS comparator groups. However, absolute comorbidity prevalence remained low across all groups. No evidence of a modifying role for hypertension, diabetes, dyslipidemia, or obesity was observed for the increased reporting of myocarditis/pericarditis among young adult males following mRNA COVID-19 vaccination. Despite the limitations inherent to spontaneous reporting systems, this study highlights a novel application of disproportionality-based approaches within VAERS to explore potential risk modifiers of vaccine-related adverse events in a hypothesis-generating manner. Rare cases of inflammation of the heart muscle (myocarditis) or of the protective layer around the heart (pericarditis), have been reported after mRNA COVID-19 vaccination, especially in young adult males. This study examined whether common conditions with cardiovascular burden—high blood pressure, diabetes, high cholesterol, and obesity—were more common among males aged 18–30 years who developed myocarditis or pericarditis after mRNA COVID-19 vaccination, which could explain the reason for an increased risk. The study used information from a US vaccine safety monitoring database (VAERS) that collects reports of health problems occurring after vaccination. Researchers compared how often these cardiovascular conditions were reported in young males with myocarditis/pericarditis after mRNA COVID-19 vaccination with how common these conditions are in vaccinated and general US populations. The study found that these conditions were reported less often than expected among myocarditis/pericarditis cases. Additional analyses also showed low reporting of these conditions across other vaccine-related adverse event reports. Overall, the findings do not suggest that high blood pressure, diabetes, high cholesterol, or obesity play a major role in the increased reporting of myocarditis/pericarditis among young adult males following mRNA-based COVID-19 vaccination. The study also highlights a new way of using vaccine safety databases to explore factors that may influence vaccine-related adverse events.
During the COVID-19 pandemic, the major burden of morbidity and mortality was in older age groups. While within-household transmission is well documented, the wider role of social networks for transmission to the older population is less well understood. To estimate whether children's birthdays were associated with grandparents' SARS-CoV-2 infection risk. This nationwide, register-based cohort study included grandparents residing in Denmark with 1 to 5 grandchildren aged 0 to 15 years during the period between February 19, 2020, and February 28, 2022. The timing of birthdays of grandchildren was used as a natural experiment to examine whether birthday-related opportunity for family gatherings was associated with the risk of SARS-CoV-2 infection in grandparents. Family structures were mapped at the individual level. Data were analyzed from June 2024 to December 2026. Birthdays of grandchildren turning 1 to 16 years. A birthday risk window, the period when positive test results could be ascribed to a celebration, was defined as the 7-day time window from 2 to 8 days after the grandchild's birthday. The primary outcome was a positive result on a SARS-CoV-2 test (by polymerase chain reaction or antigen testing) recorded in the national surveillance system. Hazard ratios comparing the hazard of having positive test result for SARS-CoV-2 (outcome) in grandparents in birthday-windows vs grandparents who at the same moment were in nonbirthday periods, were estimated using Cox proportional hazards regression with birthdays as time-varying covariates. Among 1 106 493 grandparents (median [IQR] age, 67 [60-73] years; 54% female), grandparents had 9.9% (95% CI, 7.9%-12.0%) higher hazard of SARS-CoV-2 infection during birthday risk windows. The hazard varied over time and by variant and was not seen during the period when Alpha dominated. The risk was greater for birthdays of preschool-aged grandchildren (adjusted hazard ratio, 1.15; 95% CI, 1.12-1.18) than during birthdays of school-aged grandchildren (adjusted hazard ratio, 1.07; 95% CI, 1.04-1.09). This cohort study of grandparents found that grandchild birthdays, despite official advice not to gather during part of the pandemic, were associated with increased SARS-CoV-2 infection risk among grandparents during much of the pandemic, with magnitude of risk varying over time and by viral variant. These results emphasize the role of culturally important events in intergenerational transmission dynamics, supporting future targeted risk mitigation around small family events.
Microglia are central regulators of Alzheimer's disease pathogenesis, but their roles cannot be reduced to a simple protective-versus-harmful dichotomy. Genetic, single-cell, and spatial studies have shown that Alzheimer 's-associated microglia occupy diverse disease-linked states shaped by amyloid plaques, tau pathology, lipid stress, complement activation, astrocyte signaling, aging, and immune genetic risk. Among the regulatory nodes controlling these states, SPI1, which encodes the myeloid transcription factor PU.1, has emerged as a key determinant of microglial identity and disease responsiveness. Human genetic studies suggest that reduced SPI1 expression may be protective, whereas experimental data indicate that excessive PU.1 suppression can impair essential microglial functions. This review examines the emerging concept that partial, plaque-associated reduction in PU.1 may enable a distinct lymphoid-like immunoregulatory microglial program marked by CD28 expression. Recent evidence suggests that PU.1-low CD28-positive microglia may restrain neuroinflammation and amyloid pathology, raising the possibility that Alzheimer's plaques induce not only inflammatory and phagocytic microglial responses, but also endogenous suppressive programs that limit tissue damage. We discuss this proposed PU.1/CD28 regulatory axis in relation to disease-associated microglia, TREM2-APOE signaling, complement-mediated synapse loss, antigen-presentation pathways, plaque-niche biology, and therapeutic microglial reprogramming. We also highlight major unresolved questions, including whether PU.1-low CD28-positive microglia are present and functional in human Alzheimer's disease, whether they are specific to amyloid-rich niches or extend to tau and mixed pathologies, and how such states could be safely manipulated without disrupting essential immune surveillance. We propose that lymphoid-like suppressive microglia represent a promising but still unproven framework for understanding protective neuroimmune regulation in Alzheimer's disease and for developing state-specific microglial therapies.
Type 2 diabetes mellitus (T2DM) increases the risk of cognitive impairment through metabolic-neurodegenerative interactions, yet the underlying neural mechanisms remain unclear. This study investigates whether glycemic control modulates the relationships among glymphatic dysfunction, cortical thinning, and cognition in T2DM, with a focus on whether glymphatic impairment is associated with chronic hyperglycemia-related neurostructural decline. T2DM patients were stratified by glycemic control (Hemoglobin A1c < 7.5% vs. ≥ 7.5%). All participants underwent neuropsychological assessments and magnetic resonance imaging (MRI) to quantify cortical thickness, choroid plexus volume (CPV), perivascular space (PVS) volume, and the diffusion tensor image analysis along the perivascular space (DTI-ALPS) index. Group comparisons, Spearman correlations, and mediation analyses were used to examine the pathways linking glycemic control, glymphatic function, and cortical structure. A total of 54 poorly controlled T2DM patients, 38 well-controlled T2DM patients, and 99 healthy controls were included. Poorly controlled T2DM patients exhibited worse cognitive performance compared with healthy controls. Both T2DM groups showed reduced cortical thickness in the insula, fusiform gyrus, and supramarginal gyrus relative to healthy controls, with insular atrophy significantly associated with enlarged CPV. Markers of glymphatic dysfunction, including enlarged CPV, increased PVS volume, and reduced DTI-ALPS index, were most pronounced in the poorly controlled T2DM group. Cortical thickness and glymphatic measures each correlated with cognitive performance. Mediation analysis indicated that CPV showed associations consistent with a mediating role in the relationship between HbA1c and left insular cortical thinning. Compared with other subgroups, in the poorly controlled T2DM group, glymphatic changes were more pronounced, and the glymphatic-cognitive associations were more evident. Furthermore, CPV showed associations consistent with a mediating role in the relationship between hyperglycemia and cortical thinning in T2DM patients. These findings suggest that the glymphatic system may serve as an associative link between systemic metabolic dysregulation and structural neurodegeneration, offering potential imaging biomarkers for early neurological risk assessment in T2DM.
Inhibition of Wnt/β-catenin signaling differentiates mesenchymal stem cells (MSCs) into hepatocytes. However, there is lack of data on whether transplanting these cells can enhance liver regeneration. Additionally, it remains unknown if the flavonoids quercetin and rutin and the clinically used drug lithium chloride (LiCl) can effectively differentiate MSCs into hepatocytes and promote liver regeneration. To address this, the rat bile duct ligation (BDL) fibrosis model was used. Male Wistar rats were surgically ligated. Liver function tests, histological analysis and cell tracking were also conducted to validate liver regeneration. Treatment with quercetin inhibited the Wnt pathway, while rutin and LiCl activated it. Hepatic differentiation was noted in three treatment groups: quercetin, quercetin with rutin and LiCl. We observed that the initial downregulation of the Wnt pathway, followed by its upregulation, facilitated the differentiation of MSCs into hepatocytes. Transplantation of quercetin-treated MSC-derived hepatocytes into the rat BDL fibrosis model resulted in complete restoration of liver function, normalization of elevated systemic liver enzymes and reduction of inflammation and fibrosis. Interestingly, quercetin-treated hepatocytes resulted in enhanced liver regeneration compared to rutin and LiCl. Finally, tracking of labeled hepatocytes confirmed their main localization in the liver. In conclusion, MSC-derived hepatocytes, generated through ex vivo treatment with quercetin, exhibit enhanced liver regeneration. These findings provide a novel ex vivo treatment strategy with flavonoids and the clinically used drug LiCl to achieve enhanced liver regeneration in vivo.
The widespread application of cadmium sulfide (CdS) nanoparticles (NPs) in various fields inevitably leads to their release into the environment, where they can negatively affect plants and, consequently, pose risks to human health through the food chain. Over the past two decades, the increasing release of NPs into soils, resulting from production, consumption and waste disposal, has raised significant environmental concerns. Current research focuses on understanding NPs interactions with plants to enable sustainable agriculture. Therefore, the present study examines how CdS NPs affect spinach growth and whether silicon dioxide (SiO2) NPs can alleviate CdS NPs induced stress. Three-week-old spinach plants were treated for 4 weeks with either 1 mg/l CdS NPs alone or in co-exposure with three concentrations (1, 20 and 100 mg/l) of SiO2 NPs. The central methods applied included measuring metal content, transmission electron microscopy (TEM) for cellular uptake visualization, analysis of biochemical stress markers and a metabolomic analysis to identify altered metabolic pathways. Key results demonstrated that SiO2 NPs, particularly at 1 and 100 mg/l concentrations, significantly reduced Cd content by up to 40.72% and enhanced the uptake of essential nutrients (Zn, Co, Ca, Fe). TEM imaging confirmed reduced cellular internalization of CdS NPs. The treatment also induced a strong antioxidant response and increased chlorophyll content by up to 170.10%. Metabolomic analysis revealed significant alterations in metabolites (53-75%), primarily affecting pathways related to amino acids, carbohydrates, oxidative phosphorylation and sulphur metabolism. SiO2 NPs effectively detoxify CdS NPs in spinach through a dual mechanism: limiting cellular uptake and biochemically triggering a robust antioxidant and metabolic defence system. This study provides critical insights into the application of SiO2 NPs as a nano-enabled strategy to enhance crop safety in heavy metal-contaminated environments.
We showed previously that breast cancer patients with certain circadian rhythm genotypes have higher incidences of toxicity when treated with radiotherapy at different times of day. This opens the way to genetically-guided chronomodulation as a cost-effective way to reduce side-effects while preserving treatment efficacy. We aimed to test whether similar time-of-day effects are observed in prostate cancer. The analysis used the multinational, prospective observational REQUITE prostate cancer cohort (n=1760). All patients received external beam radiotherapy and were followed for two years. Regression analyses were performed in patients with complete data including genotypes and radiotherapy fraction times (n=877). LASSO and logistic regression models incorporated genotypes of SNPs in circadian rhythm genes, time of radiotherapy and their interaction. Primary endpoints were late rectal bleeding and urinary incontinence. Multivariable models show a significant effect of genotype, time of treatment and an interaction between PER3 genotypes and time for both primary endpoints. The 44% of patients with rs696305 C/C genotype are predicted to reduce their risk of rectal bleeding from 11% when treated at 09:00 to 5% at 17:00. Similarly, the analysis predicts a significant reduction in risk of urinary incontinence from 15% to 5% by avoiding treatment in the middle of the day for patients heterozygous for rs172933 (36% of patients). These results agree with the earlier findings in breast cancer radiotherapy patients. Interaction between genotype and time of treatment suggests groups of patients could reduce side-effects and improve quality-of-life through chronomodulation.
Pathological mitochondrial fission driven by hyperactivation of the Drp1-MiD49 interaction contributes majorly to several cardiovascular diseases. Existing synthetic Drp1 inhibitors often lack selectivity between pathological and physiological mitochondrial fission, leading to poor bioavailability and off-target effects. Allosteric modulation therefore represents a more promising strategy to fine-tune Drp1 activity than complete inhibition and our study investigated whether natural antioxidants could function as selective allosteric modulators of Drp1 by targeting a characterized allosteric site at the dimer interface using molecular docking, long-timescale (1000 ns) molecular dynamics simulations and MMPBSA analysis. Docking analysis demonstrated favourable binding of the selected phytochemicals at the Drp1 allosteric interface, with Astaxanthin exhibiting the highest docking score, while Baicalin, Luteolin, and Resveratrol formed stable interactions with critical interface residues. Molecular dynamics simulations revealed that these compounds remained consistently associated with the allosteric site and promoted compact conformations of Drp1 characterized by reduced RMSD, radius of gyration (Rg) and solvent-accessible surface area (SASA) relative to apo-Drp1. RMSF, secondary structure and principal component analyses further demonstrated ligand-induced conformational transitions in functionally important regions including the G1/P-loop, G3/Switch II, G5/G-cap and the 80-loop and progressive restriction of conformational space for the Baicalin-, Luteolin-, and Resveratrol-bound complexes. MM-PBSA calculations identified Baicalin as the most energetically favourable complex, followed by Resveratrol. Independent replica simulations of the Baicalin-, Resveratrol-, and Mdivi-1-bound complexes further supported the reproducibility of the observed molecular dynamics trends. Collectively, these findings highlight the potential of Baicalin, Resveratrol, and Luteolin as promising Drp1 allosteric modulators, supporting future development of mitochondrial fission-targeted cardiac therapeutics.
Foot pain and symptoms affect 42% of pregnant women and extend beyond gestation, but the associations between foot morphology and these symptoms are not well understood. We sought to identify changes in foot shape throughout pregnancy and postpartum, using a novel multiplane digital camera-based system and to determine whether these alterations are associated with clinically important changes in the Patient-Reported Outcomes Measurement Information System (PROMIS) Pain Interference and Physical Function scores. (1) How does foot width, length, dorsum height, Arch Height Index, and medial foot area change throughout pregnancy and postpartum? (2) Are there relationships between foot shape and changes in foot shape over time with clinically important changes in PROMIS Pain Interference and Physical Function scores during pregnancy and postpartum? Between December 2023 and November 2024, we recruited patients scheduled for an obstetrics visit at Penn Medicine clinics in the Philadelphia, PA, USA, area who were between the gestational ages of 6 to 15 weeks, older than 18 years of age, had a BMI less than 45, had no history of connective tissue disorders or trauma to the lower extremities, and could understand and provide written consent. Of 2067 contacted patients, 40 consented and scheduled a laboratory visit; 78% (31 of 40) had complete datasets and were available for analysis. Pregnant participants (mean ± SD age of 35 ± 4 years, mean ± SD BMI of 68.8 ± 11.8 kg) visited the laboratory between 9 and 15 weeks (early pregnancy), 23 and 27 weeks (mid-pregnancy), and 34 and 39 weeks (late pregnancy) of gestation as well as between 9 and 15 weeks postpartum for foot shape assessment and completion of PROMIS scores. We developed a multiplane digital imaging approach designed to capture the top and medial views of the foot simultaneously to allow rapid quantification of foot shape while eliminating exposure to ionizing radiation. We validated this system by longitudinally monitoring foot shape in a control group of 18 nonpregnant participants (mean ± SD age of 28 ± 4 years, mean ± SD BMI of 62.7 ± 6.4 kg) assessed 12 weeks apart. This control group included female patients who had no previous pregnancies, were older than 18 years of age, had a BMI less than 45, had no history of connective tissue disorders or trauma to the lower extremities that resulted in medical intervention, and could provide written consent. We collected PROMIS Pain Interference and Physical Function score assessments during pregnancy and postpartum. Both scores are standardized T-scores and range from 20 to 80, with higher Pain Interference and lower Physical Function scores indicating worse outcomes. Minimum clinically important differences for pain interference and physical function in foot and ankle orthopaedics range between 5.5 and 9.8 T-score points; therefore, we used a conservative threshold of > 10, which represents 1 SD, to identify clinically important changes in pain interference and physical function during pregnancy. We found that foot shape changed from early to late pregnancy, with median (range) increases in left foot width (9.14 cm [8.42 to 11.50] versus 9.30 cm [8.55 to 10.57], median difference +0.21 cm [95% CI 0.06 to 0.25]; p = 0.02), left dorsum height (6.22 cm [5.52 to 7.12] versus 6.33 cm [5.67 to 7.78], median difference +0.17 cm [95% CI 0.01 to 0.46]; p = 0.03), and right dorsum height (6.30 cm [5.51 to 7.58] versus 6.50 cm [5.52 to 7.64], median difference +0.23 cm [95% CI 0.13 to 0.38]; p = 0.008). There were mean ± SD increases in left foot length (24.37 ± 1.24 cm versus 24.66 ± 1.12 cm, mean difference +0.29 cm [95% CI 0.09 to 0.49]; p = 0.03), left medial foot area (83.08 ± 7.12 cm2 versus 87.30 ± 7.15 cm2, mean difference +4.22 cm2 [95% CI 2.19 to 6.25]; p = 0.001), and right medial foot area (84.02 ± 7.71 cm2 versus 88.10 ± 8.53 cm2, mean difference +4.08 cm2 [95% CI 2.21 to 5.94]; p < 0.001). In addition, median (range) left foot width increased from early pregnancy to the postpartum period (9.14 cm [8.42 to 11.50] versus 9.42 cm [8.29 to 10.77], median difference +0.10 cm [95% CI 0.09 to 0.37]; p = 0.01). We found associations between foot shape and patient-reported pain during pregnancy. Pregnant participants with clinically important increases in pain interference from early to late pregnancy had increased right medial foot area during late pregnancy (median [range] 11.37 cm2/shoe size [9.72 to 15.97], median difference +1.02 cm2/shoe size [95% CI 0.42 to 1.73]; p = 0.005) and postpartum (11.26 cm2/shoe size [9.88 to 15.47], median difference +0.91 cm2/shoe size [95% CI 0.04 to 2.57]; p = 0.01) compared to the control group of nonpregnant participants (10.35 cm2/shoe size [8.76 to 11.54]). Pregnant participants with clinically important increases in pain interference from early to late pregnancy also demonstrated an increase in right foot length from early to late pregnancy when compared with nonpregnant participants who had no changes in pain interference (0.49 ± 0.59 cm versus -0.14 ± 0.49 cm, mean difference 0.63 cm [95% CI 0.21 to 1.06]; p = 0.005). Pregnant participants with clinically important declines in physical function had a greater increase in right medial foot area from early to late pregnancy than pregnant participants without a physical function decline (6.62 ± 4.94 cm2 versus 1.98 ± 4.00 cm2, mean difference 4.64 cm2 [95% CI 1.14 to 4.64]; p = 0.01). This study identified potential morphological risk factors for musculoskeletal pain and reduced function in pregnant patients. Patient-reported pain and function were predominantly associated with pregnancy-induced lower extremity edema. These findings have potential clinical impact to guide interventions (such as compression socks, massage, or foot elevation) aimed at managing foot changes during pregnancy to reduce musculoskeletal pain and disability.Level of Evidence Level III, prognostic study.
Sleep deprivation is a recognized risk factor for neuroinflammatory and neurodegenerative disorders. Dopamine signaling through D2 receptors (DRD2) has emerged as a potential immunomodulatory pathway in the central nervous system. The present study investigated whether activation of DRD2 by quinpirole (QUIN) modulates astrocytic and microglial responses and NF-κB nuclear translocation in a murine model of rapid eye movement sleep deprivation (RSD). Male CD1 mice were subjected to 72 h of RSD and treated with QUIN (2 mg/kg/day). GFAP, Iba-1, and NF-κB expression were evaluated in hippocampal subregions (CA1, CA3, dentate gyrus) and the medial parietal cortex using immunofluorescence and confocal microscopy. RSD increased GFAP and Iba-1 expression and induced morphological changes consistent with glial activation. Notably, RSD increased NF-κB nuclear expression in microglia. QUIN administration reduced Iba-1 expression, attenuated microglial morphological alterations, and reduced NF-κB nuclear expression across all analyzed regions, even in RSD-subjected mice. These findings suggest that DRD2 activation exerts anti-inflammatory effects in the brain during REM sleep deprivation and that dopaminergic signaling may represent a key target for neuroinflammation associated with sleep loss.
HIV reservoir size and decay represent distinct dimensions of viral persistence, yet whether they are governed by shared or separable immunological mechanisms during early antiretroviral therapy (ART) remains unclear. In this study, we employed multiparameter flow cytometry and bulk RNA sequencing to analyze longitudinal immune profiles across 21 treatment-naïve people living with HIV before ART initiation and at 1 and 5 months thereafter. Our findings revealed an apparent dissociation between HIV-1 DNA levels and decay rates in peripheral blood, and the two indicators appear to be relatively independent dimensions of viral persistence. Specifically, lower HIV-1 DNA levels were associated with higher frequencies of cytotoxic and adaptive-like natural killer (NK) cell subsets, whereas faster HIV-1 DNA decay was linked to restored HIV-specific CD4+ and CD8+ T-cell responses during treatment. Notably, transcriptomic analyses uncovered divergent gene expression signatures related to B cell-associated immunity and type I interferon pathways, with individuals with higher HIV-1 DNA levels exhibiting elevated expression of immunoglobulin and interferon-stimulated genes, while faster decay correlated with enrichment of antiviral and complement-related genes. Collectively, these findings provide a preliminary characterization of immune correlates of peripheral blood total HIV-1 DNA dynamics in the early phase following ART initiation. This work offers potential immune clues for exploring the viral reservoir and generates testable hypotheses for validation in future large cohorts.
Galectin-3 (Gal-3) is a β-galactoside-binding lectin implicated in metabolic inflammation, cardiovascular and renal dysfunction, neurodegenerative disorders, and obesity-related pathologies. Although Gal-3 is recognized as a clinically relevant biomarker, the mechanisms controlling its tissue expression and circulating abundance remain poorly defined. O-GlcNAcase (Oga; encoded by Mgea5), the enzyme that removes O-linked β-N-acetylglucosamine (O-GlcNAc) from proteins, regulates nutrient-sensitive signaling and transcriptional processes that overlap with Gal-3 associated disease pathways. To investigate the relationship between metabolic status and Gal-3 expression, male mice were fed a high-fat diet (HFD) for eight weeks to induce obesity. HFD-fed mice exhibited significant increases in body weight and fasting and fed blood glucose levels compared with lean controls, confirming metabolic dysregulation. ELISA revealed approximately threefold higher serum and plasma Gal-3 concentrations in obese mice, indicating enhanced Gal-3 production in diet-induced obesity. To determine whether Oga regulates Gal-3 expression, Oga wild-type (WT), heterozygous (HET), and knockout (KO) mice were analyzed. Circulating Gal-3 protein levels were significantly reduced in Oga KO mice, with intermediate levels in Oga HET animals. RT-qPCR revealed genotype-dependent modulation of Gal-3 (Lgals3) mRNA expression across multiple tissues, demonstrating tissue-specific regulation by Oga. These findings establish Oga as a critical regulator of Gal-3 expression and systemic abundance. The data reveal a mechanistic link between O-GlcNAc signaling enzyme Oga, and lectin-mediated metabolic inflammation, suggesting that Oga activity influences Gal-3 homeostasis and may affect its interpretation as a biomarker in metabolic disease.
Daily, we make visuospatial self-perspective judgments from our actual perspective (SPJ). However, social contexts often require imagining another viewpoint-visuospatial perspective-taking (VPT). VPT is costlier than SPJ, and gets even costlier in autism, schizophrenia, and aging. One explanation is interference from self-related representations. This interference may reflect a mechanism that processes various self- and other-related representations, supported by the temporoparietal junction (TPJ). Moreover, reportedly other-related representations also influence SPJ, though this remains debated. An additional explanation for VPT difficulty in literature is the mental body-schema transformation needed to adopt another perspective at greater angular disparities between perspectives. However, studies often confounded angle with left/right representational incongruence between perspectives. In our previous iEEG study, we found robust TPJ activity during both SPJ and VPT, possibly reflecting the processing of self-other representations. To test whether self-other representations are processed during both perspective judgments, we analyzed behavioral data from the same participants. We categorized trials by an object's left/right congruence between the perspectives. We hypothesized that if self-other representations are processed during both judgments, participants will show greater difficulty in incongruent vs. congruent scenarios during SPJ and VPT. Linear mixed model analysis confirmed this: for both SPJ and VPT, errors were higher on incongruent than congruent trials. These findings show that self-other representations are processed in both SPJ and VPT, and incongruent ones cause interference, highlight left/right congruence as a key factor in VPT studies, and motivate further research with a similar approach on TPJ's role in processing multimodal self-other representations.