Search Results (29,154)

In pigs, the establishment of pregnancy depends on tightly coordinated molecular interactions between the conceptus and the maternal endometrium during the peri-implantation period. Adipokines, peptide hormones produced by adipose tissue, function as endocrine mediators linking metabolic status with uterine function. Omentin-1, a hormone belonging to the adipokines group, is hypothesized to play a potential role in regulating female reproductive functions through its influence on uterine functions. Therefore, this study investigated the effects of omentin-1 on endometrial progesterone and oestradiol secretion using radioimmunoassay, on the abundance of key steroidogenic proteins and Akt phosphorylation using Western blot, and on endometrial cell proliferation and apoptosis using flow cytometry. Results showed that its actions are characterised by promotion of progesterone-dominant conditions, selective modulation of steroidogenic pathways, activation of Akt signalling, and cell-type-specific regulation of proliferation and apoptosis. This combination of effects positions omentin-1 as a previously unrecognised regulator of endometrial adaptation and suggests that adipokines may represent an important mechanistic link between metabolic status and reproductive success. Full article

[Background] Cotton seed protein is an important seed quality trait with considerable utilization potential; however, the genetic mechanisms governing its developmental dynamics remain incompletely understood. [Methods] Field trials were conducted during the 2024 and 2025 growing seasons. Using 11 upland cotton parental lines and 36 F₁ hybrid combinations obtained through Griffing-II double-cross hybridization, three planting configurations were evaluated (three-row (FR3), four-row (FR4), and six-row (FR6) film mulch configurations). Samples were harvested at eight developmental stages: 10, 15, 20, 25, 30, 40, 50, and 60 days after anthesis. Cotton seed protein content was determined using the Kjeldahl method, and protein accumulation parameters were estimated using a logistic growth model. Unconditional and conditional genetic analyses were conducted using a developmental ADM model that incorporated genotype–environment interactions. [Results] Cotton seed protein accumulation exhibited an S-shaped trend: it first increased, entered a plateau phase, then declined slightly, and rose gradually in the later developmental stage. Genotype, year, planting configuration, and their interactions significantly affected protein content and growth parameters but did not alter the underlying developmental trajectory. Among the planting configurations, FR4 exhibited the highest protein accumulation rate and maximum protein accumulation, with the lowest trait variation, and showed the most favorable overall performance under the conditions tested. Genetic analyses indicated that the dominance–environment and maternal–environment interactions were the principal components of protein accumulation, whereas additive, dominance, and maternal main effects were significant only at specific developmental stages. In terms of the genetics of cotton seed protein, the predicted genetic effects across different environments and developmental stages indicated that P6 exhibited the highest stability of genetic effects, P7 and P9 showed moderate stability, whereas P3 had relatively low stability of genetic effects. [Conclusions] The developmental pattern of cotton seed protein accumulation was largely governed by genetic effects and remained relatively stable, whereas environmental conditions mainly modified the magnitude of accumulation. For breeding aimed at improving cotton seed protein, the genetic backgrounds of parental lines should be considered together with environmental effects on cotton seed protein accumulation. Full article

Both RIT1 and RIT2 are members of the RAS superfamily of small GTPases, which regulate various cellular processes. RIT1 is widely expressed, whereas RIT2 is primarily found in neuronal tissues. Dysregulation of these proteins has been associated with several human diseases, including Noonan syndrome, cancer, Parkinson’s disease, autism, and schizophrenia. Although RIT1 and RIT2 are often compared to classical RAS proteins, they exhibit distinct regulatory and biochemical properties. Here, we demonstrate that RIT1 differs from classical RAS in GTPase cycling. Unlike classical RAS proteins, RIT1 did not respond to SOS1-mediated nucleotide exchange or p120GAP-stimulated GTP hydrolysis under cell-free conditions. These results imply that RIT1 may depend on regulatory mechanisms that differ from those of classical RAS proteins. However, the relevant physiological regulators remain unknown. Disease-associated RIT1 mutations cluster around the P-loop and Switch II regions. In this transient overexpression screening system, however, these mutations had only a modest effect on the canonical MAPK, PI3K/AKT, and JNK signaling pathways in HEK293T overexpression experiments. This suggests the existence of additional context-specific effectors and regulatory factors. We demonstrate that RIT1 and RIT2 interact with membrane lipids via a basic C-terminal extension. The KRLK-containing region contributes to the binding of phosphatidylserine and phosphoinositides. Charge-reversal mutations disrupt lipid interactions and liposome binding, supporting the functional importance of this region. In a reconstituted liposome system, galectin-3 and LZTR1, but not galectin-1, reduced the interaction of GDP-loaded RIT1 and RIT2 with liposomes. These results suggest that accessory proteins may influence RIT membrane interactions. However, their cellular relevance requires further validation. Together, our findings provide biochemical insights into RIT GTPase regulation and its interactions with membrane lipids under cell-free conditions. Full article

Type 2 diabetes (T2D) is a metabolic syndrome frequently associated with obesity and endoplasmic reticulum stress-mediated inflammation, which can trigger the unfolded protein response (UPR), impair insulin signaling, and promote apoptosis. To identify potential natural therapeutic candidates, this study investigated the mechanisms of action of 14 compounds from Uncaria tomentosa (UT), a medicinal plant from the Amazon rainforest, using in silico modeling. The study focused on the UPR, TRAF2/JNK pro-inflammatory signaling pathway, and insulin signaling pathways, which play key roles in T2D. Some of the UT compounds were docked against several human proteins involved in these pathways, and molecular dynamics simulations confirmed stable interactions between the target proteins (PERK, TRAF2, JNK, TNF-α, IRS-1, PI3K, AKT, GSK3β, and PPARγ) and four of the UT compounds, 5-Carboxystrictosidine, Cinchonain, Epicatechin and Mitraphylline. Additionally, absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties analyses were conducted to predict the four compounds, revealing suitable pharmacokinetic properties. These findings suggest that specific UT compounds may be used in experimental tests to whether investigate their therapeutic potential in managing T2D by modulating signaling pathways related to the conditions UPR, inflammation, and insulin resistance. Full article

Background: Death-associated protein kinase 1 (DAPK1) is a key regulator of apoptosis and immune responses; however, its prognostic significance in oral cancer remains insufficiently characterized. This study investigated the prognostic relevance of DAPK1 in oral squamous cell carcinoma (OSCC) and examined its associations with immune infiltration and apoptosis-related signaling pathways. Methods: A retrospective translational study design was employed, integrating TCGA-based expression and methylation analyses of 528 head and neck squamous cell carcinoma (HNSCC) tumors, UALCAN epigenetic profiling, GeneMANIA protein–protein interaction mapping, TIMER 2.0 immune correlation analyses in 422 HPV-negative HNSCC patients, and multiplex immunofluorescence validation using a tissue microarray cohort of 82 patients with histologically confirmed OSCC, of whom 75 were eligible for the final analysis at Kaohsiung Veterans General Hospital, Taiwan. Results: In vitro validation using Western blot analysis in FaDu cells showed that epidermal growth factor receptor (EGFR) inhibition with gefitinib induced upregulation of DAPK1 protein expression at 10 μM and increased total caspase-3 expression. Higher DAPK1 signal in whole-field quantification was associated with increased CD4+ and CD8+ T-cell infiltration and enrichment of apoptosis-related pathways. Patients with high DAPK1 expression demonstrated a consistent protective trend for overall survival in a pre-specified fully adjusted primary model (adjusted HR = 0.51, 95% CI: 0.21–1.21, p = 0.126), and exhibited significantly improved survival in a secondary parsimonious model (adjusted HR = 0.41, 95% CI: 0.18–0.91, p = 0.029). Multiplex immunofluorescence further confirmed stronger DAPK1 and caspase-3 staining, along with denser lymphocytic infiltration within the tumor microenvironment. Conclusions: Collectively, these findings suggest that DAPK1 is associated with apoptosis-related signaling, increased immune-cell infiltration, and favorable clinical outcomes in OSCC, although its independent prognostic value requires validation in larger cohorts. Full article

Medemia argun is a rare desert palm rich in phenolic compounds with promising biological activities; however, the instability and poor compatibility of phenols in dairy systems limit their functional application. This study characterized the phenolic composition and biological activities of M. argun phenolic extract (MAPE), developed sodium alginate–whey protein concentrate (SAlg/WP) hydrogel beads for phenolic encapsulation, and evaluated their application in fortified yoghurt. HPLC identified quercetin and gallic acid as the predominant phenolic compounds. Antioxidant, antimicrobial, antiviral, and molecular docking analyses were performed, while SAlg/WP–MAPE beads were characterized by encapsulation efficiency analysis, FTIR, and scanning electron microscopy. Fortified yoghurt containing different concentrations of encapsulated MAPE was evaluated for color, syneresis, pH, and antioxidant activity during 14 days of refrigerated storage. MAPE exhibited strong antioxidant and broad-spectrum antimicrobial activities against bacterial and fungal strains, as well as promising antiviral activity against hepatitis A virus (HAV), with an IC₅₀ of 104.89 μg/mL and a selectivity index of 1.6. Molecular docking revealed favorable interactions of rutin, chlorogenic acid, and rosmarinic acid with the HAV 3C protease. SAlg/WP beads achieved 78.92% encapsulation efficiency, while FTIR and SEM confirmed successful phenolic entrapment. Encapsulation effectively preserved MAPE functionality and significantly enhanced yoghurt antioxidant activity during storage, demonstrating its potential for functional dairy applications. Full article

Acute liver injury (ALI) is a severe clinical syndrome characterized by systemic inflammation and mitochondrial oxidative stress. This study investigated the hepatoprotective effects of paeonol (PA) in LPS/D-GalN-induced ALI and explored the potential involvement of macrophage mitochondrial function and NDUFS7. Proteomic analysis showed that the expression of mitochondrial respiratory chain-related proteins was suppressed in the liver of ALI mice, while PA treatment was associated with increased expression of several of these proteins. Integrated metabolomic analysis further indicated that PA modulated metabolites associated with energy metabolism and redox processes. In vitro experiments suggested that inflammatory RAW264.7 macrophages contributed to hepatocyte injury, whereas PA attenuated inflammatory responses and reduced macrophage-mediated injury to AML12 cells. In RAW264.7 cells, PA attenuated ROS accumulation and modulated multiple mitochondrial functional parameters, including mitochondrial membrane potential, ATP levels, the NAD⁺/NADH ratio, and complex I activity. Metabolic studies in human liver microsomes and primary human hepatocytes identified three major PA glucuronide metabolite products with five possible structural assignments. Molecular docking and microscale thermophoresis further indicated that several proposed PA metabolite structures exhibited more favorable predicted interactions and stronger binding to recombinant NDUFS7 than the parent PA. Collectively, these findings support the hepatoprotective effects of PA and its modulation of macrophage mitochondrial function, as well as the potential involvement of PA metabolites and NDUFS7, while further functional validation is required to establish their causal roles. Full article

Background/Objectives: Congenital anomalies of the kidney and urinary tract (CAKUT) are a leading cause of pediatric kidney disease. Epithelial–mesenchymal transition (EMT), governed by SNAI1 and TGF-β, and a vascular–mesenchymal program marked by PECAM1 and VIM are central to nephrogenesis, but whether these programs are transcriptionally activated in human CAKUT is unknown. We assessed their expression and spatial organization in human fetal kidneys. Methods: We reanalyzed public transcriptomic datasets for six transcripts (SNAI1, TGFB1–3, PECAM1, VIM) and performed quantitative double immunofluorescence for four gene products (SNAIL, TGF-β1, CD31, vimentin) on formalin-fixed human fetal kidneys (20 controls, 19 CAKUT), with colocalization quantified by Pearson’s coefficient. Results: No statistically significant difference in transcript or protein abundance was detected between control and CAKUT kidneys, in either the cortex or the medulla, and abundance did not change across developmental phases. In contrast, spatial colocalization of SNAIL–TGF-β and of CD31–vimentin was increased in CAKUT kidneys. Conclusions: These findings may suggest that, in human fetal CAKUT, EMT and vascular–mesenchymal programs are not quantitatively upregulated but instead show altered spatial organization of otherwise unchanged gene products. As colocalization reflects spatial proximity rather than molecular interaction, these observations are correlative and warrant functional validation. Full article

Mycobacterium tuberculosis (M. tb) possesses a unique, lipid-rich cell envelope that is critical for virulence, drug resistance, and persistence. Here, we identify the PPE family protein PPE57 as a key regulator of mycolic acid transport and host lipid exploitation. PPE57 physically interacts with the essential lipid transporter MmpL3, promoting trehalose monomycolate (TMM) translocation, enhancing trehalose dimycolate (TDM) synthesis, and increasing cell wall lipid content. Site-directed mutagenesis identified G175 as a critical residue for PPE57-MmpL3 binding. Deletion of PPE57 reduces cell wall thickness, alters lipid composition, and impairs biofilm formation. Mechanistically, PPE57 facilitates bacterial cholesterol acquisition, and during infection, activates the host PPAR-γ pathway in macrophages, leading to enhanced cholesterol uptake, lipid droplet accumulation, and increased intracellular triglyceride and cholesteryl ester levels. These changes provide a nutrient-rich niche that promotes bacterial survival and persistence. In a mouse model of infection, PPE57 deficiency results in reduced bacterial burden, milder lung pathology, and diminished lipid droplet-positive cell accumulation in lung tissues. These findings establish PPE57 as an essential accessory component of the MmpL3 lipid transport system, bridging mycobacterial cell wall assembly with host nutrient exploitation during persistent infection. Full article

Plant protein-derived bioactive peptides have attracted increasing interest as potential ingredients for health-promoting functional foods because of their reported cardiometabolic, antioxidant, anti-inflammatory, immunomodulatory, antimicrobial, gastrointestinal, and satiety-related activities. However, the field remains dominated by peptide discovery, in silico prediction, enzyme-inhibition assays, simulated digestion, and preclinical models, whereas successful translation into clinically supported and technologically viable food products is still limited. This review critically examines the gap between mechanistic promise and functional food implementation. It integrates evidence on plant protein sources, peptide-generation strategies, structure–activity relationships, gastrointestinal stability, intestinal transport, local gut activity, food-matrix interactions, processing effects, encapsulation, sensory constraints, human efficacy, regulatory substantiation, commercial feasibility, and consumer acceptance. Particular attention is given to the limited predictive value of isolated in vitro activity when peptides are exposed to digestion, epithelial barriers, complex food matrices, realistic processing conditions, and achievable dietary doses. The review also highlights that systemic absorption is not the only relevant pathway, as selected peptides may act locally within the gastrointestinal tract. Overall, the evidence indicates that peptide discovery should not be treated as the principal endpoint of research. Future progress will require translation-oriented development in which bioactivity, digestion stability, matrix compatibility, sensory quality, manufacturing reproducibility, realistic intake, human evidence, and regulatory credibility are evaluated as interdependent criteria. The most promising plant-derived peptides will therefore be those that retain sufficient activity and acceptability under real conditions of food production and consumption. Full article

Background/Objectives: Polyglutamine (polyQ) expansion induces protein misfolding, aggregation, and neurodegeneration; however, effective strategies for aggregate clearance remain limited. This study evaluated the effect of metformin, resveratrol, and curcumin on polyQ proteotoxicity using an engineered Dictyostelium discoideum model expressing 80Q repeats, and investigated their autophagy-related mechanism. Methods: In this study, 80Q-expressing Dictyostelium discoideum (D. discoideum) cells were treated with metformin, resveratrol, or curcumin. Cell viability and proliferation were assessed by growth curve and doubling-time analyses. Developmental assays examined restoration of multicellular morphology. PolyQ aggregation was analyzed using brightfield microscopy, fluorescence microscopy, Thioflavin T (ThT), and Congo Red assays. Quantitative real-time polymerase chain reaction (qRT-PCR) evaluated AMP-activated protein kinase (AMPK) and autophagy-related gene expression. Molecular docking and molecular dynamics (MD) simulations were performed to assess interactions between the compounds and AMPK. Results: All three compounds significantly improved viability and reduced doubling time in 80Q-expressing cells. Treated strains displayed improved developmental morphology and culmination compared to untreated controls. Thioflavin T (ThT) and Congo Red assays demonstrated a marked reduction in amyloid-like polyQ aggregates following treatment. Gene-expression analysis revealed the upregulation of AMP-activated protein kinase (AMPK) and core autophagy genes, indicating activation of an AMPK-linked pro-autophagy pathway involved in aggregate clearance. Computational studies further confirmed stable, energetically favorable interactions of metformin, resveratrol, and curcumin with AMPK. Conclusions: Metformin, resveratrol, and curcumin effectively reduce polyQ-associated proteotoxicity in D. discoideum and are associated with modulation of AMP-activated protein kinase (AMPK) signaling and autophagy-related pathways. These findings support the therapeutic potential of metabolic modulators against protein aggregation and establish D. discoideum as a valuable platform for screening autophagy-targeted interventions relevant to neurodegenerative diseases. Full article

Dual inhibition of p21-activated kinase 4 (PAK4) and nicotinamide phosphoribosyltransferase (NAMPT) has emerged as a promising therapeutic strategy due to its ability to simultaneously target oncogenic signaling and cellular metabolism in cancer. While existing inhibitors such as KPT9274 and PF-3758309 have demonstrated preclinical activity, their clinical translation has been limited by poor selectivity, suboptimal efficacy, and dose-limiting toxicities. Reliance on a small number of available compounds is therefore insufficient, highlighting the need for systematic optimization to identify candidates with improved therapeutic profiles. Although several dual PAK4 and NAMPT inhibitors such as GNE2861, LCH7749944, and PF3758309 have been identified in our previous study, the number of compounds available is limited. Identification of new candidate compounds would allow researchers to identify those that are the most efficient, those that are the most potent and selective, and those that have optimal pharmacokinetics. In this study, we performed additional large-scale drug screening to identify new candidate dual PAK4 and NAMPT inhibitors and expand the diversity of this inhibitor class. Computational molecular docking was used to evaluate binding affinities toward both targets, followed by drug–protein interaction analyses to assess binding stability and interaction patterns at the molecular level. Several new candidate compounds demonstrated favorable predicted binding to both PAK4 and NAMPT, with distinct interaction profiles compared to previously reported inhibitors. Biochemical assays further demonstrated inhibitory activity against both PAK4 and NAMPT among several selected compounds, supporting their potential as dual-target inhibitors. These findings provide mechanistic insight into dual-target engagement and highlight structural features associated with improved binding behavior. Expanding the pool of dual inhibitors enhances opportunities for preclinical development, supports optimization of pharmacokinetic and safety profiles, and strengthens datasets for drug discovery. Collectively, this work broadens the landscape of dual PAK4 and NAMPT inhibitors and supports their potential application across multiple cancer types beyond triple-negative breast cancer. Full article

Metabolic dysfunction-associated steatotic liver disease (MASLD) is characterized by excessive liver lipids and metabolic imbalance. Thyroid hormone receptor alpha (THRA) is essential for liver lipolysis; however, its transcription function is dependent on successful nuclear localization. Since natural polyphenols are known to modulate THRA activity, making them potential candidates for MASLD management, their roles in THRA nuclear trafficking have not been elucidated. In this study, an integrative computational method was utilized to investigate the impact of polyphenol binding on THRA nuclear import. Protein–protein docking simulations suggested that polyphenol binding may augment the interaction of THRA and importin-α, while importin-7 was predicted to be essential for basal transport. In conclusion, our findings showed that polyphenol binding could potentially support THRA nuclear import, thus potentially implicating a new process in MASLD. Full article

The utilization of protein-rich agricultural by-products offers a sustainable strategy for developing plant-based meat analogs. This study investigated the effects of cold-pressed hemp seed cake (HSC) incorporation at levels of 0–20% on the pasting, thermal–rheological, textural, and structural properties of high-moisture meat analogs produced by extrusion. Increasing HSC incorporation significantly reduced peak viscosity, indicating altered starch–protein–fiber interactions within the blends. During temperature-sweep measurements, all formulations exhibited elastic-dominant behavior, with the storage modulus remaining higher than the loss modulus throughout heating and cooling. Although the initial viscoelastic moduli decreased with increasing HSC content, the final moduli after cooling were comparable to those of the control. Incorporation of 15% and 20% HSC significantly decreased hardness from 44.02 to 38.88 N and chewiness from 1730.20 to 1517.31 g, whereas springiness, cohesiveness, and the hardness degradation ratio remained largely unchanged. Fibrous structures were maintained in all formulations, while cutting strength tended to increase and the texturization degree numerically increased from 1.04 to 1.15 at 20% HSC. These findings demonstrate that HSC can replace up to 20% of the conventional protein–starch blend while maintaining thermal viscoelasticity and anisotropic fibrous structure, although producing a moderately softer high-moisture meat analog. Full article

Roseofungin, a polyene macrolide antibiotic produced by Streptomyces roseoflavus, was evaluated for antiviral activity against Influenza A virus using complementary in vitro, in ovo, and preliminary in vivo toxicity models, supported by molecular docking and molecular dynamics simulations. The compound exhibited antiviral activity against multiple influenza A virus subtypes, including an oseltamivir-resistant H1N1 strain, with EC₅₀ values ranging from 2.0 to 2.5 μg/mL and selectivity indices of 20–25. Functional assays demonstrated that roseofungin interfered with early stages of the viral life cycle by reducing viral adsorption and partially inhibiting neuraminidase activity. In virucidal assays, treatment with roseofungin resulted in a 1.5–2.5 log₁₀ reduction in infectious viral titers. Acute toxicity studies in mice indicated low toxicity under the experimental conditions (LD₅₀ > 25 mg/kg). Molecular docking predicted favorable interactions of roseofungin with multiple influenza A virus proteins. Subsequent molecular dynamics simulations of the hemagglutinin–roseofungin complex maintained the stability of the predicted binding mode across three independent 50 ns simulations (150 ns cumulative). Collectively, these findings demonstrate that roseofungin possesses broad-spectrum antiviral activity against genetically diverse Influenza A virus strains and represents a promising lead compound for the development of novel membrane-active antiviral agents. Further studies are warranted to elucidate its molecular mechanism of action and to evaluate its therapeutic potential in vivo. Full article