Search Results (2,820)

Background/Objectives: The mechanisms leading to oxidative stress and cellular dysfunction during stroke are not well understood. Methods: We tested if transient cerebral artery occlusion (MCAo) in mice results in the generation of oxidized phospholipids (OxPLs) that contribute to neuronal cell death and glial activation. Results: Both in vitro and in vivo cerebral ischemia and reperfusion injury (IRI) resulted in the elevation of specific OxPLs. Neuronal cell death was determined in the presence of OxPLs and the natural OxPL E06 antibody (antioxidized phospholipid antibody) protected the cells from the toxic effects. IRI in mice gave rise to increased immunoreactivity of OxPLs in the brain. E06 reduced inflammatory markers in the brain following IRI, including iba-1, GFAP and inflammatory cytokines. In addition, OxPLs gave rise to M1 and Mox microglial phenotypes which was reversed in the presence of E06 and elicited a more M2 phenotype. Nrf2-deficient mice showed increased infarct volumes and microglia from Nrf2^−/− mice showed a reduction in Mox gene expression, and E06 protects both mice and cells from the Nrf2-deficit. Finally, AB1-2 Ab which recognizes the E06 Ab, ameliorates the impact of E06 Ab on infarct volume in the mouse model. Conclusions: Taken together, the data indicate that OxPLs play an important role in inflammation and neuronal cell loss in cerebral IRI and inactivation of OxPLs may provide novel targets for potential drug targets in the treatment of stroke. Full article

Artisanal colonial cheese (ACC) produced from raw milk is a rich reservoir of autochthonous lactic acid bacteria (LAB), but strain-level evidence supporting safe downstream application and technological stability remains limited. In this study, 10 LAB isolates from ACC were screened for phenotypic safety, antimicrobial susceptibility, and probiotic-related traits, and their viability was further assessed after inulin-based spray-drying microencapsulation under different storage temperatures. All isolates showed no hemolytic or mucinolytic activity and did not produce gelatinase, supporting an initial safety profile, and all strains were sensitive to at least two antimicrobial classes. Strain prioritization identified Lacticaseibacillus casei LAB06, LAB09, and LAB10 and Lactiplantibacillus plantarum LAB03 as the most robust candidates for downstream development because they maintained stable cell counts throughout simulated gastrointestinal digestion. Inulin spray-drying yielded structurally stable microcapsules and supported refrigerated storage, with substantially lower viability losses at 4 °C than at 25 °C; notably, L. plantarum LAB01 and LAB02 showed the best refrigerated shelf-life, remaining above 6.0 log CFU/g after 45 days. Together, these results position ACC as a source of promising LAB candidates and highlight cold-chain-compatible microencapsulation as a strategy to support safe functional food development with potential public health benefits. Full article

Leucine-rich repeats and immunoglobulin-like domains protein 1 (Lrig1) is a functional inhibitor of the epidermal growth factor receptor. Lrig1-positive stem cells are located in the isthmus region of the mouse hair follicle (HF) and are known contributors to sebaceous gland (SG) formation and homeostasis. In this study, we performed a topical tamoxifen inducible diphtheria toxin-mediated ablation of Lrig1-expressing cells in transgenic mice to investigate their function in vivo. Selective depletion of Lrig1-positive cells resulted in a complete but reversible loss of SGs, with atrophy beginning at day 14 and full recovery occurring after six months. In the absence of the Lrig1 niche, junctional-zone keratinocytes adopted an interfollicular epidermis-like phenotype (K1-positive), and repopulating cells from other epidermal compartments failed to differentiate into the sebocyte lineage. These findings demonstrate that Lrig1-positive progenitors are crucial for proper sebaceous gland morphogenesis and maintenance. Our results highlight the importance of Lrig1-positive cells in SG-related skin physiology. Full article

Background/Objectives: Type 2 diabetes (T2D) and obesity are chronic metabolic disorders associated with cognitive impairment and neuronal damage. The hippocampus, a region sensitive to nutrient excess, is critical for integrating sensory and metabolic signals. This study aimed to determine the early onset of cognitive and motor deficits induced by obesity and/or hyperglycemia and to characterize associated hippocampal alterations in Zucker Diabetic Fatty (ZDF) rats. Methods: Male ZDF rats (13 weeks old) were categorized into three groups: lean control, obese normoglycemic (ZDF-NG), and obese hyperglycemic (ZDF-HG). Assessments included zoometric parameters (weight and adiposity), biochemical assays (glucose tolerance, insulin response, and lipid profile), and behavioral tests (Open Field and Novel Object Recognition). Hippocampal health was evaluated through stereological neuronal density analysis and redox balance markers. Results: Both obese groups exhibited significant visceral adiposity and hyperlipidemia. The ZDF-HG group was further characterized by glucose intolerance, hepatic insulin resistance, and reduced β-cell function. Behavioral results showed that while obesity decreased motor activity, hyperglycemia significantly exacerbated the loss of both short- and long-term recognition memory. Histologically, obesity was associated with decreased neuronal density in the hippocampal DG and CA1 regions. Furthermore, hippocampal ROS was significantly elevated in the ZDF-HG group, and glutathione reductase activity was reduced in both obese phenotypes. Conclusions: The findings demonstrate that obesity initiates hippocampal neurodegeneration and motor decline, and that hyperglycemia severely impairs recognition memory. These results emphasize the critical interplay between metabolic dysfunction and cognitive decline, highlighting the necessity of managing both obesity and T2D to prevent early neurodegenerative changes. Full article

Background/Objectives: Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra (SN). Because current therapeutics have limited efficacy once PD is fully developed, it is crucial to start disease-modifying interventions during the prodromal stage of PD. In the present study, we aimed to evaluate whether intranasally delivered human umbilical cord mesenchymal stem cells (hUC-MSCs) have an efficacy in the rotenone-induced prodromal PD-like phenotype mouse model. Methods: To produce the prodromal PD mouse model, C57BL/6 mice were treated with intraperitoneal (i.p.) rotenone for 1 or 2 weeks. hUC-MSCs or PBS were delivered intranasally for 1 or 2 weeks with rotenone injection. We subsequently performed behavioral assessments to evaluate motor and non-motor features, followed by pathological analyses of the mouse brains. Results: Intranasal administration of hUC-MSCs restored motor performance and protected dopaminergic neurons in the SN of mice treated with rotenone for 2 weeks. In the 1-week rotenone mice, hUC-MSCs treatment ameliorated depressive-like behaviors and attenuated olfactory dysfunction. Furthermore, intranasal hUC-MSC treatment suppressed the accumulation of protein aggregates in the brains of mice, which is associated with enhanced autophagic function, as indicated by increased LC3B and normalization of LAMP2A protein expression. Conclusions: Our data demonstrate that intranasal administration of hUC-MSCs improves non-motor symptoms at early time points and attenuates progression to nigrostriatal loss and motor deficits in the rotenone-induced PD mouse model. These findings support the potential of a non-invasive, prodromal-stage intervention to modulate early pathological progression in PD. Full article

This study evaluated the effect of rearing conditions with outdoor access on carcass traits, technological meat quality, and muscle fibre morphology in broiler chickens. Hubbard JA757 broilers were reared until 81 days of age under either control or experimental conditions. Both groups were housed indoors under identical management until day 35; thereafter, the experimental group had access to an outdoor paddock, while the control group remained indoors. At 81 days of age, birds were slaughtered, carcass composition was recorded, and breast (pectoralis major) and thigh (biceps femoris) muscles were analyzed for pH, colour, cooking loss, and shear force. Muscle fibre characteristics were assessed histochemically. Control birds showed higher slaughter and chilled carcass weights, whereas experimentally reared birds exhibited higher carcass yield, lower breast proportion, higher thigh proportion, and reduced abdominal fat. Experimental rearing was associated with higher L* values, higher shear force, and lower cooking loss in both muscles. In the pectoralis major, fibre density was higher in controls, while fibre diameter was higher in the experimental group. In the biceps femoris, the experimental group showed a higher proportion of oxidative βR fibres and larger fibre cross-sectional areas. These results demonstrate that the rearing system influences carcass composition, technological meat properties, and muscle fibre characteristics, leading to distinct phenotypic and technological differences between the groups. Full article

Deciphering gene and protein functions and interactions remains a core challenge in biology and medicine. Gene set analysis and multi-omics tools are widely used to interpret gene lists; however, they often overlook shared evolutionary patterns among genes. These conservation and loss patterns, shaped by billions of years of evolutionary pressure, can uncover co-evolutionary signals within gene sets, yet they remain frequently underexplored. In this study, we apply normalized phylogenetic profiling (NPP) across 1905 eukaryotic species and introduce CladeOScope-GSA, a tool for analyzing user-defined gene sets. CladeOScope-GSA uncovers common signatures of conservation, revealing whether a gene set evolves as a cohesive unit or as distinct co-evolving submodules. By tracing gene set origins, diversification, and shared evolutionary histories, the tool identifies the structural organization and key components of gene networks, exposing functional similarities, phenotypic associations, and broader biological relationships. We demonstrate its utility through two well-characterized cases: the porphyria-related pathway and the dynein gene family. In both, CladeOScope-GSA recapitulates known functional substructures and uncovers previously unrecognized evolutionary insights, underscoring its value for advancing our understanding of gene function and pathway evolution on a broad scale. Full article

To maintain the integrity of the outer membrane of Gram-negative bacteria, such as Escherichia coli, the levels of two essential components, phospholipids (PL) and lipopolysaccharide (LPS), are tightly regulated, although the underlying molecular mechanisms are unclear. E. coli synthesizes three main PLs, including essential phosphatidylethanolamine and phosphatidylglycerol and nonessential cardiolipin (CL). We showed that CL synthesis is conditionally essential in ΔlapD bacteria. Using this synthetic lethal phenotype, we isolated suppressors that rescued growth at elevated temperatures. We showed that loss-of-function mutations in cdsA encoding CDP-diglyceride synthetase, and pgsA, which encodes phosphatidylglycerophosphate synthase, bypass this lethality. Such mutations reduce the relative abundance of acidic phospholipids, which are otherwise elevated in Δ(lapD clsA) bacteria, and increase the amounts of cis-vaccenic acid without altering amounts of LpxC mediating the first committed step in LPS biosynthesis. Interestingly, overexpression of genes, including accC and glnB, whose products can inhibit fatty acid/PL synthesis, overcame the lethality of Δ(lapD clsA) bacteria. We demonstrated that PgsA co-purifies with LapB, which regulates LpxC stability and acts as a hub for proteins involved in PL and LPS biosynthesis, including LapD. Overall, our results reveal that LapD is positioned at the regulatory nexus between LPS assembly and fatty acid/PL synthesis. Full article

Prader–Willi syndrome (PWS) is a rare imprinting-related neurodevelopmental disorder caused by loss of paternally expressed genes within the chromosome 15q11–q13 region, including SNORD116, MAGEL2, and NDN. It provides a natural model for examining how genomic imprinting disruptions shape neural development and psychiatric vulnerability. This review synthesizes current evidence to clarify the mechanistic pathways linking imprinting defects and epigenetic dysregulation to neuropsychiatric outcomes in PWS. Published studies—including patient-derived induced pluripotent stem cell (iPSC) models, animal knockout systems (e.g., Magel2-null models), transcriptomic and DNA methylation datasets, and human neuroimaging research—were identified through targeted searches of PubMed and Web of Science and integrated narratively rather than through systematic procedures. Across these data sources, deletion-type PWS is primarily associated with impaired neuronal maturation, altered serotonergic signaling, and locus-specific transcriptional dysregulation. Maternal uniparental disomy (mUPD) is characterized by broader epigenetic alterations within the imprinted domain, genome-wide transcriptional effects, dopaminergic pathway alterations, and disrupted prefrontal–limbic connectivity linked to increased psychosis risk. Importantly, available evidence supports substantial phenotypic and mechanistic overlap between PWS subtypes, with genotype–phenotype associations reflecting probabilistic tendencies rather than categorical distinctions. Collectively, convergent findings across molecular, neurochemical, and systems-level studies support a mechanistic continuum extending from imprinting defects to behavioral phenotypes. These insights position PWS as a translational model for understanding how epigenetic dysregulation contributes to psychiatric risk and highlight the need for genotype-informed, mechanistically grounded research to advance biomarker development and targeted therapeutic strategies. Full article

Background: The tumor microenvironment (TME) plays a central role in pancreatic ductal adenocarcinoma (PDAC) progression, yet how mechanical cues shape stromal cell behavior remains poorly defined. Here, we investigate how dimensional priming of adipose-derived stromal cells (ADSCs) alters their immunomodulatory functions and subsequent impact on PDAC growth. Methods: ADSCs were cultured under two-dimensional (2D) or three-dimensional (3D) conditions and evaluated using in vitro co-culture systems with PDAC organoids and in vivo xenograft models. Stromal phenotype, cytokine secretion, tumor growth, invasion, and immune cell infiltration were assessed. Results: ADSCs cultured in three-dimensional (3D) hydrogels exhibited reduced Caveolin-1 (CAV-1) expression and reprogramming toward a stress-adapted, CAF-like phenotype compared with two-dimensional (2D) cultures. In vitro, 2D-primed ADSCs constrained PDAC organoid growth, increased MMP-2 activity, and required direct cell–cell contact to suppress tumor viability. By contrast, 3D-primed ADSCs preserved organoid structure but markedly enhanced tumor cell migration through soluble factors, accompanied by increased IL-6 and TNF-α and reduced IL-10 secretion during co-culture. In vivo, 3D-primed ADSCs promoted the largest tumors with aggressive invasion and loss of Col-Tgel containment associated with tumor expansion, whereas 2D-primed ADSCs suppressed tumor growth and maintained gel boundaries. Immunohistochemistry confirmed elevated Ki-67 in tumors containing 3D-primed ADSCs, while macrophage infiltration (F4/80⁺) was highest in 2D-primed tumors and lowest in 3D-primed tumors. Conclusions: Dimensional priming fundamentally reprograms ADSC phenotype and alters their stromal–immune interactions, generating a tumor-permissive state that accelerates PDAC progression. These findings identify mechanical cues as critical regulators of stromal plasticity and highlight dimensional priming as a potentially targetable axis within the PDAC microenvironment. Full article

This study systematically investigated the genomic alterations in Saccharomyces cerevisiae driven by Replication Factor A (RFA) dosage insufficiency using a promoter-replacement strategy combined with mutation accumulation and whole-genome sequencing. Our findings reveal that transcriptional suppression of RFA2 or RFA3 leads to severe growth inhibition. RFA deficiency induces a distinct mutational spectrum characterized by a high frequency of monosomy and terminal deletions, indicative of severe replication stress. Furthermore, loss of heterozygosity is significantly enriched at centromeres and high-GC regions, underscoring the role of RFA in stabilizing intrinsic genomic barriers. Utilizing an APOBEC3B-induced mutagenesis assay, we demonstrate that RFA insufficiency leads to the extensive accumulation of exposed ssDNA with a distinct bias towards the lagging strand template. Notably, we observed that cells spontaneously inactivate Mismatch Repair (MMR) genes, such as MSH2 and PMS1, to survive RFA-induced stress. This hypermutant phenotype grants a certain degree of growth recovery on Low Galactose (LG) medium. Overall, these findings demonstrate that RFA dosage is a key determinant of genomic integrity and elucidate how repair pathway modulation drives adaptive evolution under replication stress. Full article

Background/Objectives: Autoimmune Polyendocrinopathy with Candidiasis and Ectodermal Dystrophy is an extremely rare autosomal recessive disorder caused by inborn errors of immunity. It is due to a loss-of-function mutation in the AIRE autoimmune regulator gene. Its manifestations include autoimmunity affecting endocrine glands, in addition to non-endocrine manifestations including dental enamel hypoplasia, alopecia areata, hepatitis, and chronic mucocutaneous candidiasis. Globally, 10 cases per million are affected by this condition, with higher incidence in highly consanguineous populations. Here, we describe a novel AIRE gene mutation in a pediatric patient from Lebanon, along with the observed phenotype. Method: A nine-year-old boy with history of craniosynostosis presented with jaundice. His past medical history was significant for recurrent oral thrush, keratoconjunctivitis, nail dystrophy, and alopecia. Upon presentation, he had jaundice, isolated splenomegaly, and severe failure to thrive. Laboratory tests showed transaminitis, cholestasis, and hypergammaglobulinemia. Abdominal ultrasound findings were suggestive of cirrhosis with compensated portal hypertension. The differential diagnosis included viral infection, inborn errors of metabolism, and autoimmune hepatitis. Results: Exome sequencing identified a novel homozygous pathogenic variant in the AIRE gene, NM_000383.4: c.1066dup p.(Arg356Profs*16), confirming the diagnosis. Conclusions: This study expands the genotypic and phenotypic spectrum of a rare inborn error of immunity in a child with chronic mucocutaneous candidiasis, enamel hypoplasia, and hepatitis. Full article

Background: Physical activity (PA) and weight regulation are influenced by both genetic and lifestyle factors. This study aimed to evaluate the predictive value of Polygenic Risk Scores (PRSs) for PA and weight outcomes, and their interaction with dietary habits. Methods: Baseline phenotypic data from 202 participants enrolled in the iMPROVE study were analyzed. The sample included 59 men and 143 women, aged 19–65 years. Based on baseline Body Mass Index (BMI), 75 participants were classified as having overweight and 126 as having obesity. Polygenic risk scores (PRSs) were calculated for 197 participants with available genetic data. PA was operationalized as metabolic equivalent of task minutes per week (MET-mins/week), derived from self-reported activity questionnaires. Weight-related outcomes included log-transformed weight loss from baseline to month 3 and change in BMI post-intervention. Interactions with diet were examined using both the randomized intervention dietary groups and previously extracted dietary patterns from the iMPROVE cohort. Correlation analyses and linear regression models were used to assess the main effects of PRSs and dietary patterns, as well as gene–diet interactions. Results: The measured PA PGS002254 presented a nominal significant interaction with diet group for weight loss post-intervention (B = 7.57, SE = 3.57 × 10⁰, p = 0.04; R² = 0.06). Similarly, the sedentary behavior PGS001923 presented a significant interaction with the “High in unsaturated fats and fruit juice consumption” pattern for baseline MET-mins/week (B = 1.51 × 10³, SE = 4.135 × 10², p = 0.001; R² = 0.091). Conclusions: Genetic predisposition influences short-term activity and weight outcomes, with dietary patterns moderating these effects. However, the multifactorial nature of lifestyle behaviors is being underscored by the modest variance explained. Full article

Background: Several studies have attempted to identify the initiating drivers of small intestinal neuroendocrine tumor (SI-NET) development and the molecular mechanisms underlying their progression and metastatic spread. Previous gene expression studies have used bulk microarrays or RNA sequencing to compare tumor tissue with normal intestinal mucosa. However, the intestine comprises multiple distinct cell types, and bulk analyses are limited by this cellular heterogeneity, which can confound tumor-specific signals. Methods: We performed single-cell RNA sequencing on primary SI-NETs and paired normal mucosa from two patients to directly compare tumor cells with their cells of origin, the enterochromaffin (EC) cells. To minimize type I errors, we applied a two-step validation strategy by overlapping differentially expressed genes with an external single-cell dataset and cross-referencing candidate genes for enteroendocrine expression in the Human Protein Atlas. Results: For further distinction and characterization, ECs were subdivided into serotonergic and non-serotonergic clusters. This analysis revealed that the SI-NET cells are transcriptionally more similar to serotonergic ECs, consistent with serum metabolite profiles derived from clinical parameters. Our analyses uncovered a loss-of-expression program characterized by regulators of epithelial differentiation and in parallel, a gain-of-expression program displayed neuronal signaling gene induction, implicating functional reprogramming toward neuronal-like properties. Together, these specific losses and gains suggest that our patient-derived SI-NETs undergo adaptation through both loss of enteroendocrine functions and acquisition of neurobiological-promoting signaling pathways. Conclusions: These findings nominate candidate drivers for further functional validation and highlight potential therapeutic strategies in our patient cohort, including restoring suppressed Notch signaling and targeting aberrant neuronal signaling networks. However, even with a two-step validation procedure, the modest cohort size limits statistical power and generalizability, particularly for the proposed association to a serotonergic phenotype. Larger, multi-patient single-cell studies are required to confirm these mechanisms and establish their clinical relevance. Full article

Background: We describe the post-mortem analysis of a CARD11 variant allele, p.Ser995Leu, identified in fraternal twins who died in early infancy with no identifiable cause of death. CARD11 variants through varied inheritance models can alter immune function through loss- or gain-of-function mechanisms, involving distinct protein domains; yet the significance of GUK domain variants remains poorly characterized. Twin autopsies showed non-specific findings, such as pulmonary macrophage accumulation and splenic white pulp expansion, but without infection or structural abnormalities. Methods: Whole-exome sequencing, performed as part of molecular autopsies, identified the shared CARD11 p.Ser995Leu variant, previously classified as a variant of uncertain significance (VUS). We assessed evolutionary conservation across CARD family proteins and species and predicted functional impact using in silico tools, which estimate the likelihood that a variant is deleterious. AlphaFold-based structural modeling emphasized qualitative biophysical assessment. Using epidemiological data, population allele frequency, and Bayesian ACMG variant classification, we assessed competing hypotheses under an autosomal dominant model. Results: The p.Ser995Leu substitution affects a conserved, surface-exposed β-sheet within the GUK domain. While CADD scores exceeded 20, other predictive algorithms offered only partial support of pathogenicity. Structural modeling suggested a potential GUK domain destabilization. Integrating genetic, pathologic, immunologic, and probabilistic modeling, we propose a biologically plausible model in which the variant, like other GUK variants, may alter NF-κB or other signaling pathways and is likely pathogenic. Conclusions: While the CARD11 p.Ser995Leu variant’s contribution to disease is uncertain without functional validation or parental testing, and phenotypic findings are non-specific, the presence of an ultra-rare GUK domain variant in both twins, combined with in silico and statistical modeling, supports its interpretation as likely pathogenic or high risk. The results highlight the challenges of data-limited post-mortem variant interpretation. Full article