Search Results (153,107)

Soybean protein isolate (SPI) gel has been demonstrated to exhibit suboptimal stability and a coarse texture. Selective enzymatic hydrolysis modification has been demonstrated to effectively enhance the functional properties and structural stability of the protein. The objective of this study was to modify SPI using alkaline protease and papain. The impact of selective enzymatic hydrolysis on SPI was examined through the analysis of hydrolysis degree (DH), particle size, and protein purity. A systematic exploration was conducted in order to investigate the structural and quality characteristics of SPI gel. Indicators such as secondary structure changes, texture characteristics, water-holding capacity (WHC), rheology, and microstructure were analyzed. The findings indicate that when the DH of the SPI solution is 1%, its particle size is reduced relative to that when DH is 0.5%. The SDS-PAGE results indicated that alkaline protease could hydrolyze most of the 7S and 11S components in SPI into shorter peptides, while papain retained more of the 7S and 11S components and generated peptides with larger molecular weights. Fourier-transform infrared (FT-IR) spectral analysis indicated that following the process of enzymatic modification, the contents of α-helix and β-sheet in the secondary structure of SPI increased, while the contents of β-turns and random coils decreased. In the context of gel performance, it has been demonstrated that papain-modified SPI, attributable to its elevated content of macromolecular peptides, manifests superior WHC, hardness, springiness, cohesiveness, chewiness, storage modulus (G), and microstructure in comparison to alkaline protease-modified gel. Concurrently, the gel performance of papain modified SPI is significantly superior to that of unmodified SPI gel. This research provides a significant theoretical foundation and practical reference for promoting the efficient application of SPI in the domain of food processing. Full article

Background: Type 2 diabetes mellitus (T2DM) is a complex metabolic disease requiring multi-targeted therapeutic strategies. Gunnera perpensa and Erythrina zeyheri are traditionally used in diabetes management, but their mechanisms remain poorly understood. Methods: This study used in vitro, metabolomics, and network pharmacology approaches to elucidate their antidiabetic potential. Leaf extracts were screened for glucose utilization in C2C12 cells, and cytotoxicity in Vero cells. Metabolites profiled via GC×GC-TOF-MS and those retrieved from Phytochemical Interaction Database were evaluated for drug-likeness and target prediction using SwissADME and SwissTargetPrediction. Diabetes-related targets were obtained from databases, and overlapping targets were used to construct interaction networks using Cytoscape and STRING. Functional enrichment analyses were conducted via DAVID for GO and KEGG pathways. Results: G. perpensa acetone and methanol extracts enhanced superior glucose utilization (IC₅₀ = 78.5 and 94.8 µg/mL, respectively), with low cytotoxicity (LC₅₀ > 600 µg/mL). Key compounds including arabinose, identified from both plants, showed multi-target binding potential against STAT3, PIK3RI and JAK2. Enrichment analyses revealed pathways related to insulin signaling, inflammation, and glucose metabolism. Conclusions: This study supports the therapeutic relevance of phytochemical synergy in the traditional use of both plants and demonstrated systems-level approaches for elucidating complex drug–target interactions in T2DM. Full article

Background/Objectives: Patients with psychotic disorders have a lifetime risk of suicide attempt (SA) of around 10 to 50%. Genetic variants in the corticotropin-releasing hormone-binding protein (CRHBP) and FK506-binding protein 5 (FKBP5) genes, which are implicated in the hypothalamic–pituitary–adrenal axis and childhood trauma (CT), are considered risk factors for SA. This study aimed to examine the interaction between the CRHBP and FKBP5 genes and CT in the development of SA. Methods: We included 350 patients, 180 patients with schizophrenia and 170 with bipolar disorder. The patients were divided into two groups: 175 with a history of SA and 175 without, and a sample of 350 healthy controls was also included. The Multifactor Dimensionality Reduction program was used to identify G × E interactions between the CRHBP (rs7728378, rs10474485, and rs1875999) and FKBP5 (rs3800373 and rs9296158) and CT in SA. Results: The analysis showed that the interaction of CRHBP and FKBP5 with CT increases the risk of presenting at least one SA (OR 4.17; 95% CI [2.67–6.52]; p < 0.0001). Additionally, we observed interaction with childhood abuse (OR 4.09; 95% CI [2.61–6.39]; p < 0.0001), mainly with emotional (OR 3.67; 95% CI [2.34–5.77]; p < 0.0001) and sexual abuse (OR 3.32; 95% CI [2.11–5.23]; p < 0.0001). Conclusions: Our research indicates that genetic variations in CRHBP and FKBP5 genes and a history of CT increase the probability of presenting at least one SA in patients with schizophrenia and bipolar disorder. Full article

This study examined how coenzyme Q10-supported high-intensity interval training (HIIT) influences plasma lactate threshold, skeletal muscle oxidative capacity, circulating irisin and corticosterone, and hippocampal brain-derived neurotrophic factor (BDNF) and glial fibrillary acidic protein (GFAP) levels in rats. Forty-eight male Sprague Dawley rats (8 weeks old; 250.4 ± 11.2 g) were randomized into four groups: control (C), coenzyme Q10 (Supp), HIIT, and HIIT with coenzyme Q10 (HIITsupp). HIIT was performed five days per week on a treadmill following a four-stage familiarization. Coenzyme Q10 (5 mg/kg/day) was given by gavage 30 min before HIIT during weeks II–IV. Plasma lactate threshold, corticosterone, irisin, and citrate synthase (CS) activity were measured by ELISA, while hippocampal BDNF and GFAP were analyzed by both ELISA and immunohistochemistry. The HIITsupp group showed greater muscle mass, CS activity, plasma irisin, and hippocampal BDNF, along with lower GFAP and lactate threshold than the C, Supp, and HIIT groups. The Supp group had the lowest corticosterone, while the HIIT group maintained the highest lactate threshold before supplementation. Principal Component Analysis (PCA) indicated distinct clustering, with the C group closely associated with GFAP and corticosterone, whereas the HIITsupp group aligned with oxidative and neurotrophic markers. Coenzyme Q10-supported HIIT improved muscle oxidative capacity, lowered lactate, and modulated corticosterone, GFAP, and hippocampal BDNF, indicating integrated metabolic and neurobiological adaptations. Full article

Introduction: Breast cancer is a heterogeneous malignancy influenced by diverse molecular profiles. The L-type amino acid transporter 1 (LAT1), encoded by the SLC7A5 gene, plays a key role in tumor metabolism, growth, and angiogenesis. Through its role in amino acid transport and activation of the mTORC1 signaling pathway, LAT1 has emerged as a potential therapeutic target. Objective: To evaluate SLC7A5/LAT1 expression and its association with clinicopathological parameters and molecular subtypes of invasive carcinoma of no special type (NST) in a Pakistani cohort. Methods: Eighty-three patients who underwent mastectomy or modified radical mastectomy for histologically confirmed primary invasive carcinoma of no special type were included. Immunohistochemistry was used to assess SLC7A5/LAT1 expression. Associations with clinicopathological features and molecular groups were analyzed using the Chi-square test. Results: The mean age of SLC7A5-positive patients were 48.4 ± 10.8 years. Overall, 24.1% of patients demonstrated SLC7A5 positivity. Although SLC7A5 expression was more frequent in cases categorized as having moderate or poor prognosis based on the Nottingham Prognostic Index (NPI), this trend was not statistically significant. Similarly, no significant associations were observed between SLC7A5 expression and other clinicopathological or molecular variables. Conclusions:SLC7A5/LAT1 expression was identified in approximately one-quarter of invasive breast carcinoma cases. Its expression appeared more common in tumors with poorer NPI categories, but without statistically verified associations. These findings suggest that SLC7A5 may act independently of conventional clinicopathological parameters. Larger, longitudinal studies with survival follow-up are required to clarify its prognostic and therapeutic significance. Full article

Background/Objectives: Copper-based wood preservatives are widely used to protect timber from fungal decay; however, the emergence of copper-tolerant fungi reduces their long-term effectiveness. This study aimed to elucidate the molecular mechanisms underlying copper resistance in Saccharomyces cerevisiae through adaptive evolution and transcriptomic profiling. Methods: A copper-resistant mutant was developed via stepwise exposure to CuSO₄·5H₂O, and its gene expression profile was compared to the wild-type strain under copper stress and non-stress conditions using Affymetrix GeneChip Yeast Genome 2.0 arrays. Results: Differential expression analysis revealed upregulation of key genes involved in copper transport (ATX1 and CTR1), the oxidative stress response (RCK1 and SOD1), and metal ion detoxification (FRE3 and SLF1). Functional enrichment analysis highlighted the significant activation of pathways related to protein folding, mitochondrial function, and transcriptional regulation. Conclusions: These findings provide insights into the adaptive strategies employed by S. cerevisiae to tolerate copper stress and suggest potential gene targets for the development of more effective wood preservatives capable of mitigating fungal resistance. Full article

Background/Objectives: Pancreatic cancer (PC) remains one of the deadliest malignancies, with challenges that hinder early detection and few actionable molecular targets. In this study, we aimed to identify biomarkers predictive of PC to support its diagnosis and treatment. Methods: Proteins from formalin-fixed, paraffin-embedded pooled samples of PC (n = 15; 5 pools) and chronic pancreatitis (n = 10; 5 pools) tissues were analyzed via label-free quantitative proteomics using liquid chromatography-tandem mass spectrometry. Immunohistochemistry (IHC) was performed on PC tissue microarrays to assess S100 calcium-binding protein P (S100P) and cathepsin E (CTSE) expression (IHC evaluable pairs: n = 78 for S100P; n = 82 for CTSE). Transwell invasion assays were conducted to evaluate the effects of these proteins on PC cell invasiveness, and Western blotting was used to validate protein expression and elucidate associated molecular mechanisms. Results: Both S100P and CTSE were overexpressed in PC tissues compared with those in adjacent normal tissues. Elevated S100P expression correlated with poor prognosis, whereas higher CTSE expression predicted favorable outcomes; both served as independent prognostic factors in PC. Functionally, S100P promoted PC cell invasion, whereas CTSE suppressed it. Mechanistically, both proteins appeared to regulate epithelial–mesenchymal transition (EMT) and invasive capacity through activation or inhibition of the phosphoinositide 3-kinase (PI3K)–protein kinase B (AKT) signaling pathway. Conclusions: Elevated expression of S100P and CTSE in PC tissues serves as independent indicators in our model of patient survival. Both proteins regulate EMT and invasion, potentially via the PI3K–AKT pathway, and hold significant promise as prognostic biomarkers and therapeutic targets in PC. Full article

Pecan is an important oilseed tree species valued for its nutrient-rich nuts. WRKY and VQ proteins play crucial roles in plant growth, development, and stress response. However, few WRKY and VQ genes in pecan have been functionally analyzed due to functional redundancy caused by gene duplication. In this study, 89 CiWRKYs and 47 CiVQs were identified in pecan genome, which were unevenly distributed across chromosomes. Gene structure and conserved motif analyses revealed high diversity among members. Duplication analysis indicated that segmental duplication was the major factor of family expansion of CiWRKY and CiVQ. Ka/Ks ratios revealed that most duplicated gene pairs underwent purifying selection. Promoter analysis identified numerous cis-acting elements associated with light response, hormone regulation, and abiotic stress, implying their potential regulatory roles in development and stress response. Expression data across six tissues demonstrated tissue-specific patterns, with several genes highly expressed in flowers and roots. Transcriptome analysis revealed that 63 CiWRKY and 27 CiVQ genes were significantly upregulated under drought stress. qRT-PCR validation confirmed that CiPaw.10G165200 and CiPaw.04G072500 were highly induced by salt treatment, with expression levels increasing over 100-fold at 8 d. Moreover, CiPaw.10G165200 was also highly expressed under ABA treatment, which indicated it might play a key role in the response to abiotic stresses. Our results provide valuable insights into the evolutionary patterns and functional roles of WRKY and VQ genes in pecan and lay a foundation for improving stress tolerance and molecular breeding in this economically important nut tree. Full article

Metabolic dysfunction-associated steatotic liver disease (MASLD) is increasing in both prevalence and severity, highlighting the need for non-invasive biomarkers to assess disease activity. Extracellular vesicles (EVs), which carry molecular cargo from their cells of origin, hold promise as accessible biomarkers. We performed proteomic profiling of plasma-derived EVs from 70 patients with MASLD to identify protein signatures associated with key histological features (steatosis, metabolic dysfunction-associated steatohepatitis (MASH), and fibrosis). These proteins were subsequently correlated with magnetic resonance (MR)-based liver imaging. Plasma EV protein profile differed between mild (S1) and advanced steatosis (S3). H4C1, OIT3, and ANPEP were elevated in S3, while CCDC25 and KLHL41 were decreased (|log₂ fold change| > 1, p < 0.05). KLHL41 had a weak-to-moderate correlation with proton density fat fraction (PDFF) (R = −0.34, p = 0.016). GP1BA was upregulated in MASH (log₂ fold change = 1.13, p = 0.03) but showed weak correlation with cT1, an imaging parameter for steatohepatitis (R = 0.22, p = 0.173). In fibrosis, complement component 7 (C7) was elevated in advanced (≥F3) vs. mild fibrosis (<F2) (log₂ fold change = 0.95, adjusted p = 0.002) and correlated with MR elastography-derived liver stiffness (R = 0.38, p = 0.004). The AUC of C7 for differentiating <F2 vs. ≥F2 and <F3 vs. ≥F3 was 0.80 (95% CI: 0.69–0.91) and 0.83 (95% CI: 0.72–0.93), respectively. In conclusion, plasma EVs contain distinct protein signatures associated with steatosis, steatohepatitis, and fibrosis in MASLD. These preliminary findings support the potential utility of plasma EVs as non-invasive biomarkers and provide insights into disease pathophysiology. However, further validation in larger, independent cohorts is necessary to confirm these associations and establish their clinical relevance. Full article

Background: Clear cell renal cell carcinoma (ccRCC) is a prevalent urological malignancy, accounting for approximately 1.6% of all cancer-related deaths in 2022. While endocrine-disrupting chemicals (EDCs) have been implicated as risk factors for ccRCC, the toxicological profiles and immune mechanisms underlying Bisphenol A (BPA) exposure in ccRCC progression remain inadequately understood. Materials and Methods: Protein–protein interaction (PPI) analysis and visualization were performed on overlapping genes between ccRCC and BPA exposure. This was followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to elucidate potential underlying mechanisms. Subsequently, 108 distinct machine learning algorithm combinations were evaluated to identify the optimal predictive model. An integrated CoxBoost and Ridge regression model was constructed to develop a prognostic signature, the performance of which was rigorously validated across two independent external datasets. Finally, molecular docking analyses were employed to investigate interactions between key genes and BPA. Results: A total of 114 overlapping targets associated with both ccRCC and BPA were identified. GO and KEGG analyses revealed enrichment in cancer-related pathways, including pathways in cancer, endocrine resistance, PD-L1 expression and PD-1 checkpoint signaling, T-cell receptor signaling, endocrine function, and immune responses. Machine learning algorithm selection identified the combined CoxBoost-Ridge approach as the optimal predictive model (achieving a training set concordance index (C-index) of 0.77). This model identified eight key genes (CHRM3, GABBR1, CCR4, KCNN4, PRKCE, CYP2C9, HPGD, FASN), which were the top-ranked by coefficient magnitude in the prognostic model. The prognostic signature demonstrated robust predictive performance in two independent external validation cohorts (C-index = 0.74 in cBioPortal; C-index = 0.81 in E-MTAB-1980). Furthermore, molecular docking analyses predicted strong binding affinities between BPA and these key targets (Vina scores all <−6.5 kcal/mol), suggesting a potential mechanism through which BPA may modulate their activity to promote renal carcinogenesis. Collectively, These findings suggested potential molecular mechanisms that may underpin BPA-induced ccRCC progression, generating hypotheses for future experimental validation. Conclusions: These findings enhance our understanding of the molecular mechanisms by which BPA induces ccRCC and highlight potential targets for therapeutic intervention, particularly in endocrine and immune-related pathways. This underscores the need for collaborative efforts to mitigate the impact of environmental toxins like BPA on public health. Full article

Background and Objectives: Age-related macular degeneration (AMD) is an age-associated retinal disorder characterized by blood–retinal barrier (BRB) breakdown and pathological angiogenesis, leading to vascular leakage. The intravitreal administration of anti-VEGF agents remains the most effective treatment for neovascular AMD. However, repetitive intravitreal injections have risks, causing side effects such as cataracts, bleeding, retina damage, and, in severe cases, post-injection endophthalmitis. Hence, the development of innovative drug delivery systems is essential to minimize the risks and discomfort associated with intravitreal injections. Materials and Methods: We developed a microemulsion (ME)-based topical drug delivery system incorporating α-linolenic acid (ALA). In brief, pseudo-ternary phase diagrams were constructed by the water titration method using different combinations of surfactants and cosurfactants (S_mix-Cremophor RH 40: Span 80: Transcutol P in ratios of 1:1.05, 1:1:1, 1:1:1.5) containing ALA as the oil phase. Three blank microemulsions (ME1, ME2, and ME3) were prepared and characterized based on the optimized pseudo-ternary phase equilibrium with a S_mix ratio of 1:1:1. Results: ME3, with an average particle size of 38.59 nm, was selected as the optimized formulation for developing drug-loaded ME containing Fenofibrate, Axitinib, and Sirolimus. The drug-loaded ME showed particle size (46.94–56.39 nm) and in vitro release displayed sustained and longer time drug release for 240 h. The irritation and antiangiogenic activities were evaluated using the hen’s egg chorioallantoic membrane (HET-CAM) assay employing the optimized ME loaded with each drug. Among the three drug-loaded ME, the Sirolimus ME showed a reduction in blood vessel sprouting in the HET-CAM assay, indicating strong antiangiogenic activity. Treatment with the optimized blank ME and Sirolimus ME significantly (p < 0.05) reduced COX-2 protein expression in LPS-stimulated RAW 264.7 cells, suggesting their potential anti-inflammatory effects. Conclusions: Overall, we suggest that the α-linolenic acid-based Sirolimus microemulsion may serve as a promising topical therapeutic approach for managing AMD and offering a potential alternative to invasive intravitreal injections. Full article

Background: Extracorporeal membrane oxygenation (ECMO) can affect the disposition of drugs, notably by sequestering them in a circuit. This review aimed to provide a comprehensive summary of existing ex vivo studies investigating the impact of contemporary ECMO circuits on drug sequestration, and to examine the associations between the physicochemical properties of drugs, the features and settings of ECMO devices, and the extent of drug sequestration. Method: A comprehensive search was conducted to identify ex vivo studies that determined drug concentrations in ECMO circuits. Studies that did not allow for the proper assessment of drug loss by degradation were excluded. Drug characteristics and experimental conditions were recorded. Drug sequestration in the circuit was calculated as the difference between the drug loss measured in the ECMO circuit and the drug loss due to spontaneous degradation measured under control conditions. To identify predictors of drug sequestration, a stepwise multiple linear meta-regression was applied by testing the physicochemical properties of drugs and ECMO device features/settings. Results: A total of 40 studies were identified, of which 21 were included in the analysis, covering 41 drugs. The Maquet membrane oxygenator was the most used brand (73%). About half of the circuits were adult and half were pediatric. Our final regression model retained lipophilicity, and to a lesser extent ionization at a physiological pH, as significant predictors of drug sequestration (R² 0.44, relative standard error 23%). Protein binding had no additional effect. Anti-infectives were the most studied class of drugs (n = 28). Antibiotics were overall not significantly sequestered, while lipophilic drugs such as posaconazole, voriconazole, paracetamol, fentanyl, sufentanil, propofol, thiopental, dexmedetomidine and amiodarone were highly sequestered (≥50%). However, this sequestration occurred mainly within the first few hours of the experiments, possibly reflecting a saturation effect. Conclusions: Lipophilic drugs are significantly sequestered in ex vivo ECMO circuits, although this effect may be limited by early saturation. Full article

Integrating positive welfare indicators into dairy science is gaining attention, yet the biological correlates of affective states in commercial herds remain poorly understood. This study explored associations between dairy cows’ emotional states quantified through the Positive Affect Index (PAI) derived from the Qualitative Behaviour (QBA) Assessment and milk biomarkers, yield, and udder health indicators across 37 commercial farms. Descriptive statistics, housing-adjusted linear regressions, and partial Spearman correlations were used to explore these relationships. Higher PAI values, indicating more positive herd-level emotional states, were significantly associated with lower SCC (22% reduction, p = 0.016) and lower odds of elevated DSCC (OR = 0.69, p = 0.002), reflecting improved udder health. Positive affect was also linked to a higher lactose concentration (p < 0.001) and an increased fat-to-protein ratio (FPR). A tendency for higher milk yield (+1.07 L per milking, p = 0.077) and slightly lower protein content was observed, consistent with a dilution effect. These associations remained robust after sensitivity analyses and were independent of housing type. The results demonstrate that milk composition and udder health biologically reflect positive emotional states, supporting the integration of behavioural assessments and milk biomarkers as a non-invasive framework for advancing welfare-oriented and precision dairy farming. Full article

Soybean meal causes health issues in piglets due to the presence of antigenic proteins. Although enzymatic treatment can break down antigenic proteins, subsequent high-temperature drying may impair amino acid (AA) digestibility via the Maillard reaction. This study evaluated whether the air-drying temperature affects the ileal AA digestibility of a novel reduced-sugar enzyme-treated soybean meal (ESM) in 25 kg pigs, using fishmeal as a high-digestibility reference. In two trials using pigs fitted with simple T-cannulas in the terminal ileum, ileal digestibility was assessed. In trial 1, a replicated 3 × 3 Latin square design with three diets (fishmeal, ESM, and a nitrogen-free diet; two pigs per diet) and three periods were used per square. Fishmeal showed greater apparent (82.50% vs. 45.01%) and standardized (86.60% vs. 48.86%) digestibility of crude protein and all amino acids than ESM dried at 130 °C. In trial 2, eight pigs were allocated to two diets in a two-period crossover design to compare the AA digestibility of ESM dried at high (130 °C; HtESM) and low (80 °C; LtESM) temperatures. LtESM exhibited greater apparent (82.24% vs. 52.40%) and standardized (86.37% vs. 56.47%) digestibility of crude protein and more amino acids than HtESM. Collectively, the drying temperature critically determined the AA digestibility of ESM, irrespective of its reducing sugar content. Full article