International Journal of Molecular Sciences最新文献

Poly(3-hydroxybutyrate) (P3HB) was used to produce biocompatible composites with hypromellose as an additive. The study aimed to assess their biological and mechanical properties, as well as specific thermal parameters and phase content. Differential scanning calorimetry was applied to analyze the phase transitions of both biocomposites and the polymer matrix. Furthermore, the thermal parameters-encompassing both non-equilibrium and equilibrium states-of the biocomposites and unfilled P3HB were evaluated according to their thermal history. Using equilibrium parameters such as the heat of fusion for fully crystalline materials and the heat capacity change at the glass transition for fully amorphous composites, we estimated the degrees of crystallinity as well as the mobile and rigid amorphous fractions. Adding hypromellose to the P3HB matrix reduced crystallinity compared to the unfilled material. At the same time, an increase in the amorphous phase was observed. It was also discovered that the rigid amorphous fraction exists solely in biocomposites containing 2% by mass of filler. Thermogravimetric analysis showed that the thermal stability of all biocomposites surpasses that of unfilled P3HB. Adding an extra 1% filler by mass raises the degradation temperature by about 37 °C compared to unfilled P3HB. The immunosafety of the tested biocomposites, with very low or no endotoxin contamination, was confirmed in accordance with Food and Drug Administration and European Medicines Agency guidelines. The study clearly demonstrates the influence of the filler in the P3HB matrix on various structural, thermal, mechanical, and biological properties of the prepared biocomposites.

Recovery of nutritional and bioactive molecules by pomegranate peel (PP) has found wide applications in food and pharmaceutical industries. We investigated protective effects of a PP extract (PPE) from Mediterranean (Mazara del Vallo, Italy) on intestinal inflammation by using in vitro and ex vivo models. Reactive oxygen species (ROS) and lactate dehydrogenase (LDH) levels, as well as tight junction protein-1 (ZO-1) expression, were determined in lipopolysaccharide (LPS)-injured Caco-2 cells treated with PPE. We evaluated anti-inflammatory and antioxidant effects of PPE in isolated colon specimens of adult male mouse (C57/BL6) stimulated by LPS. Cyclooxygenase-2 (COX-2), nuclear factor-kB (NF-kB), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), as well as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX), and inducible nitric oxide synthase (i-NOS) gene expression was determined. We also characterized phytochemical composition of the extract through chromatographic (HPLC-UV) and spectrophotometric techniques. PPE was rich in punicalagins A and B, along with other polyphenols such as hydroxytyrosol (HT), catechin, p-coumaric acid, and rosmarinic acid. In Caco-2 cells, PPE reduced ROS generation and LDH release, restoring intestinal barrier integrity by upregulating ZO-1 expression. In addition, PPE increased SOD, CAT, and GPX and suppressed COX-2, NF-kB, TNF-α, IL-1β and i-NOS LPS-induced gene expression in colon. PPE mitigates inflammation and oxidative stress, restoring intestinal barrier function. The beneficial effects induced by the extract could be related to the pattern of polyphenolic composition, with particular regard to HT, rosmarinic acid, p-coumaric acid, catechin, as well as punicalagins A and B.

3,4-Dihydroxybenzaldehyde (DBD) is a polyphenolic active constituent derived from Gastrodia elata. Its characteristic phenolic structure is associated with diverse bioactivities, such as anti-inflammatory, antioxidant, and cardioprotective effects. However, its role and underlying mechanisms in combating Alzheimer's disease (AD) remain inadequately elucidated. In this study, we employed computational and experimental approaches to investigate the anti-AD effects of DBD. Molecular dynamics simulations revealed that DBD binds to Aβ fibrils via π-π stacking, hydrophobic interactions, and hydrogen bonds, suggesting its potential to disrupt Aβ fibril stability and thereby inhibit aggregation. In vivo experiments in an AD C. elegans model demonstrated that 2 mM DBD treatment significantly delayed paralysis and extended lifespan. It also improved locomotor activity and pharyngeal pumping rates, while reducing lipofuscin accumulation. These results collectively suggest that DBD promotes healthspan-associated phenotypes. Broad-targeted metabolomics analysis indicated that DBD significantly altered the metabolic profile of the worms. Further mechanistic investigations suggested that the protective effects of DBD are associated with the activation of the DAF-16/FOXO and SKN-1/Nrf2 signaling pathways, accompanied by enhanced resistance to oxidative and thermal stress in nematodes. These findings suggest that DBD exhibits anti-AD potential through multimodal mechanisms, which involve interference with Aβ toxicity and reinforcement of cellular defense. This study supports DBD as a candidate compound and provides a rationale for its further investigation.

Type 2 diabetes and insulin resistance are increasingly recognized as conditions influenced not only by genetic and lifestyle factors but also by infectious and microbial exposures. Diarrheagenic pathogens, including enterotoxigenic, enteroaggregative, and enterohemorrhagic Escherichia coli, as well as other enteric microorganisms, disrupt the gut microbiota and compromise intestinal barrier integrity. These alterations promote dysbiosis, increased intestinal permeability, and systemic exposure to lipopolysaccharides and other microbial products, leading to metabolic endotoxemia and chronic low-grade inflammation. In parallel, pathogen-induced modulation of host immune responses contributes to adipose tissue inflammation, mitochondrial dysfunction, and impaired insulin signaling. This review summarizes current evidence linking diarrheagenic pathogens to insulin resistance, with emphasis on the microbiota-immune-metabolism axis. Understanding these interactions highlights novel perspectives on the pathogenesis of insulin resistance and suggests that targeted modulation of the gut microbiota or reduction in pathogen-driven inflammation may represent therapeutic opportunities to improve metabolic outcomes.

Migraine is a complex neurovascular disorder in which immune signaling intersects with vascular and neural circuits, yet the tissue and cell-type context of common genetic risk remains incompletely defined. We integrated large-scale migraine genome-wide association study (GWAS) summary statistics with Genotype-Tissue Expression (GTEx) v8 expression and splicing quantitative trait loci (eQTLs and sQTLs), Bayesian co-localization, single-cell RNA sequencing of peripheral blood mononuclear cells (PBMCs) from migraine cases and controls, a healthy single-cell multi-omics atlas (assay for transposase-accessible chromatin (ATAC) plus RNA), high-dimensional weighted gene co-expression network analysis (hdWGCNA), and embryo-level spatial transcriptomics. Genetic signals were enriched in peripheral arteries, heart, and blood, and gene-level enrichment highlighted mucosal-smooth muscle organs including the bladder and the cervix endocervix. Cell-type prioritization consistently implicated endothelial and vascular smooth muscle lineages, with additional support for inhibitory interneurons and bladder epithelium. In PBMC T cells, co-expression modules capturing cytotoxic/activation and T-cell receptor signaling programs contained migraine-prioritized genes, including PTK2B, nominating immune activation circuitry as a component of genetic susceptibility. Spatial projection further localized risk concordance to craniofacial/meningeal interfaces and visceral smooth muscle-mucosal structures. Together, these analyses delineate a systemic neuroimmune-vascular architecture for migraine and provide genetically anchored candidate pathways and targets for mechanistic and therapeutic follow-up.

Paternal deprivation has behavioral, neurochemical, and neuroendocrine consequences in adulthood. Socially monogamous prairie voles (Microtus ochrogaster) raised only by the mother (monoparental care, MP) showed low levels of alloparental behavior and delayed pair bonding formation in adulthood compared to those raised by both parents (biparental care, BP). However, the effects of paternal deprivation on adult neurogenesis and the epigenetic mechanisms involved remain to be elucidated. Here, we focused on the impact of MP rearing on neural stem cells (NSCs) proliferation under basal conditions and in response to cohabitation with the sexual partner during pair bonding formation. At basal conditions, we found a significant decrease in the number of new proliferative NSCs (BrdU⁺/SOX2⁺) in male and female MP voles compared to BP animals in the subventricular (SVZ) and subgranular zone (SGZ). After 24 h of cohabitation, in MP males, there was an increase in the number of newborn cells in the SVZ but not in the SGZ. However, this increased proliferation was lower than in BP males. In females, we did not observe significant differences compared to controls. Finally, we evaluated the enrichment of H3K4me3 (activation) and H3K27me3 (silencing) epigenetic marks in the new cells, finding differences between rearing systems and sexes.

Circulating tumor DNA (ctDNA) has gained increasing attention as a non-invasive biomarker with potential utility across multiple stages of melanoma. ctDNA reflects tumor-derived genetic alterations in real time and has shown value in detecting minimal residual disease, identifying early recurrence, estimating tumor burden, and monitoring response to systemic therapies. In early-stage melanoma, postoperative ctDNA positivity is strongly associated with higher recurrence risk and often precedes radiologic detection. In advanced disease, ctDNA correlates with tumor volume and can distinguish responders from non-responders during targeted therapy and immunotherapy, while also identifying emerging resistance mechanisms. Despite these advantages, clinical implementation remains limited by low shedding in early-stage disease, variation among detection platforms, and the absence of standardized clinical thresholds. Recent advances, including fragmentomics, methylation assays, and multi-target sequencing strategies, aim to improve sensitivity, particularly in low-tumor-burden settings. Integration of ctDNA with radiomics, artificial intelligence, and digital pathology represents an additional opportunity to enhance precision in risk stratification and treatment adaptation. This review summarizes current evidence on ctDNA biology, detection methods, and clinical applications in melanoma and outlines ongoing challenges and future directions required for translation into routine practice.

Restriction-modification (RM) systems contribute to genome plasticity in Mycoplasma hominis, a facultative pathogen with an extremely small but highly heterogeneous genome. The MhoVII RM system, which contains a fusion of two methyltransferases (MTases), M1 and M2, was recently identified within a family of Type II RM systems, but its specificity and biological function remained unknown. Phylogenetic analysis revealed that M1 and M2 belong to distinct MTase classes clustering within the YhdJ and MTaseD12 branches, respectively. In this study, the dissemination, expression and function of the MhoVII system was analyzed in detail using Oxford Nanopore-based methylation analysis, recombinant expression of the individual RM components in Escherichia coli, and methylation-sensitive restriction assays. It was thus possible to demonstrate that M1 and M2 methylate the complementary non-palindromic motifs GATG and CATC, and that the associated restriction endonuclease cleaves only DNA lacking 6mA methylation at these sites. The transcriptional analysis of mid-to-late logarithmic cultures indicated a polycistronic organization of the MhoVII genes, and GATG/CATC-driven methylation analysis revealed culture-dependent methylation differences, suggesting a post-transcriptional regulation, whereas in the infection of HeLa cells, MhoVII transcription was highest at the beginning and was then gradually downregulated in the later stages of infection. These findings establish MhoVII as a previously uncharacterized Type II RM system.

Globally, antibiotic resistance is a growing concern, motivating the search for alternatives. Bovine mastitis is an inflammatory disease of the udder caused by various microorganisms, many of which are resistant to various antibiotics, impacting the quality of dairy products and farmer income. In this study, the in vitro bioactivity of the methanol leaf extract, fractions (ethyl acetate (CeEtOAc), butanol (CeBuOH), hexane (CeHx), dichloromethane CeDCM), and water (CeAq), and a purified compound, quercetin-3-O-rhamnoside isolated from the CeEtOAc fraction of Combretum elaeagnoides Klotzsch, were investigated against six Staphylococcus aureus (S. aureus) strains isolated from clinical cases of bovine mastitis and two reference ATCC strains (S. aureus ATCC 29213 and S. epidermidis ATCC 35984). Methods used for assessing bioactivity included serial microdilution for antibacterial efficacy, crystal violet staining and p-iodonitrotetrazolium (INT) metabolic assays for anti-biofilm activity, and a microdilution assay for anti-quorum-sensing potential. The anti-inflammatory assays included 15-lipoxygenase enzyme inhibition and nitric oxide assays. Cytotoxicity screening was conducted using a tetrazolium-based colorimetric assay against bovine dermis cells. The extracts and fractions exhibited moderate to good antibacterial activity with minimum inhibitory concentration (MIC) values ranging from 0.07 to 1.04 mg/mL, with the ethyl acetate fraction being the most effective. The anti-biofilm activity of the extract, fractions, and isolated compound (quercetin-3-O-rhamnoside) varied at time zero (T0), with inhibition ranging from 3% to 100%. The CeDCM and CeEtOAc fractions exhibited the most potent anti-biofilm effects after 24 h, with inhibition ranging from 24% to 91%. The extracts and fractions exhibited significant inhibition (>50%) of biofilm within the incubation times (T0-T48), and quercetin-3-O-rhamnoside alone had >60% inhibition at 48 h. The CeEtOAc fraction had the most significant anti-quorum-sensing activity (IC₅₀ < 0.08 mg/mL). The methanol extract and fractions exhibited significant anti-inflammatory activity, inhibiting nitric oxide production (IC₅₀: 7-26 µg/mL). In contrast, the CeAq, CeHx, and CeDCM fractions showed the best inhibitory activity against the 15-lipoxygenase enzyme (IC₅₀ = 3-4 µg/mL). The extracts and fractions were non-cytotoxic to bovine dermis cells (LC₅₀ = 0.88-1 mg/mL). Combretum elaeagnoides extract and its fractions are recommended for further investigation as potential herbal treatments for the management of mastitis and its symptoms.

The realization of efficient and durable earth-abundant electrocatalysts for alkaline hydrogen evolution reaction (HER) is critical for scalable hydrogen production, yet remains limited by insufficient intrinsic activity. Herein, we demonstrate a precursor-controlled hydrothermal strategy that enables precise morphology and surface-state regulation of spinel Co₂NiO₄ directly grown on nickel foam, allowing a clear correlation between catalyst architecture and HER performance. By replacing urea with hexamethylenetetramine, an ultrathin, highly interconnected two-dimensional nanosheet network (CNO-HT) is obtained, which promotes efficient electron transport, rapid electrolyte penetration, and maximized exposure of catalytically active sites. Structural and spectroscopic analyses confirm the formation of phase-pure cubic Co₂NiO₄ with enriched mixed-valence Ni and Co species, favoring enhanced redox activity. The CNO-HT catalyst exhibits a low overpotential (86 mV at 10 mA cm^-2) and a smaller Tafel slope (103 mV dec^-1), significantly outperforming the urea-derived counterpart. Importantly, the catalyst maintains stable HER operation for 96 h at both 10 and 100 mA cm^-2, with post-stability electrochemical analyses confirming preserved kinetics and interfacial properties. This work establishes precursor-regulated nanosheet engineering as general and scalable strategy to unlock the intrinsic catalytic potential of spinel metal oxides, offering actionable design principles for next-generation non-noble electrocatalysts for alkaline hydrogen production.