Search Results (1,648)

Background: Goldenhar syndrome (oculo–auriculo–vertebral spectrum, OAVS) is a rare congenital disorder characterized by craniofacial malformations, systemic anomalies, and significant phenotypic variability. Although it is the second most common craniofacial malformation after a cleft palate, the genetic etiology of Goldenhar syndrome remains largely unexplored. This study aimed to identify genetic variants contributing to Goldenhar syndrome in a Lebanese family with three affected individuals, using whole-exome sequencing and complementary genomic approaches. Methods: Whole-exome sequencing was performed on the nuclear family to identify variants associated with the syndrome. Complementary DNA methylation and gene ontology analyses were conducted to explore epigenetic modifications. Results: A missense shared variant in the MID1 between the affected individuals [NP_000372.1): p. Ile593Phe] gene was observed in the family, while current ACMG evidence was insufficient to establish causality. Additional variants were identified, including a de novo mutation in FBXW11 and a rare frameshift alteration in NDUFAF8, with limited segregation, implicating these genes in associated phenotypes such as craniofacial anomalies and cardiac defects. DNA methylation analysis revealed hypomethylation at CpG sites within the ZC3H3 gene, suggesting an epigenetic contribution to disease variability. Conclusions: Our findings underscore the genetic and epigenetic complexity of Goldenhar syndrome, providing new insights into its molecular etiology and highlighting the challenges of variant interpretation in familial cases of rare congenital disorders. Full article

The high potential of saliva for use in the non-invasive diagnosis of a number of diseases raises a number of questions regarding the substantiation of normal and abnormal salivary composition criteria. Factors that must be considered when forming patient cohorts include age, hormonal status, and circadian variability. However, the influence of sex remains controversial. The aim of this study was to investigate the influence of sex on the biochemical composition of normal saliva, including amino acid and lipid profiles, cytokine levels, and electrolytes. The study involved 120 healthy volunteers (75 females and 45 males). The amounts of electrolytes (NH₄⁺, K⁺, Na⁺, Mg²⁺, Ca²⁺, Cl⁻, SO₄²⁻, NO₂⁻, NO₃⁻, F⁻, PO₄³⁻), amino acids (Arg, Lys, Tyr, Phe, His, Leu+Ile, Met, Val, Pro, Thr, Ser, Ala, Gly), cytokines (VEGF, MCP-1, TNF-α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-18, INF-α, INF-γ), and biochemical parameters (protein, urea, total content of α-amino acids, imidazole compounds, lipid peroxides) were analyzed. Lipid content was determined based on the intensity of absorption bands at 1396, 1458, 2853, 2923, and 2957 cm⁻¹ in the IR spectra of salivary lipid extracts. A clear sex correlation was found for amino acid and lipid content in saliva. For electrolytes and biochemical parameters, median differences were demonstrated in some cases; however, the range of variation for all parameters overlapped. Although overall cytokine profiles did not show clear multivariate separation, significant differences between sexes were observed for individual cytokines (IL-1β and IL-10). A comprehensive assessment of all parameters (amino acids, lipids, cytokines, etc.) allows for the formation of a sex-associated metabolic profile of saliva. Therefore, it is recommended to avoid the use of mixed cohorts when analyzing the amino acid and lipid profiles of saliva. Full article

Background: Pyroptosis, a pro-inflammatory programmed cell death process, is a key player in tumor biology, including in triple-negative breast cancer (TNBC). Inhibiting G9a has been proven to exert anticancer effects; however, the molecular mechanism of the effects remains unclear. The study aimed to illustrate whether inhibiting G9a can suppress the process of TNBC cells by promoting pyroptosis and investigate the underlying mechanisms. Methods: MCF-10A, MDA-MB-231 and SUM159PT cell lines were used for in vitro study. CCK8 and EdU staining assay were used to examine the cell proliferation, and flow cytometry assay was performed to evaluate cell death. Inflammatory factors were measured by ELISA kits. The mRNA and protein expression levels were analyzed by qRT-PCR, Western blot, and immunofluorescence staining. Transmission electron microscopy was used to observe the morphological changes in cells. Results: We found that knockdown of G9a suppressed the growth and the abilities of invasion and migration, induced pyroptosis, and increased the expression of RIG-I, p-STAT1, and GSDME of TNBC. Furthermore, a RIG-I inhibition Cyclo (Phe-Pro) partially rescued the activation of pyroptosis enhanced by knockdown of G9a. Conclusions: These findings indicate that inhibiting the function of G9a induces pyroptosis in TNBC cells by the RIG-1/STAT1/GSDME pathway, which provides a new therapeutic target for TNBC treatment. Full article

The variegated cutworm Peridroma saucia Hübner, a recently emerged polyphagous pest in China’s Huang-Huai River Basin, uses sex pheromones (Z)-11-hexadecenyl acetate (Z11-16: Ac) and (Z)-9-tetradecenyl acetate (Z9-14: Ac) for mate finding. Insect pheromone-binding proteins (PBPs) serve as the primary filter for detecting specific sex pheromones. Although comprehensive functional analyses of PBPs exist, their binding mechanisms remain poorly characterized. In this study, we elucidated the binding properties and mechanisms of PsauPBP3 in sex pheromone recognition by computational and experimental approaches. PsauPBP3, predominantly expressed in male P. saucia antennae, showed high binding affinity for both Z11-16: Ac and Z9-14: Ac, as demonstrated by binding-free-energy calculations and fluorescence binding assays. Molecular dynamics simulations and docking studies identified five key residues (Thr-10, Phe-13, Ile-53, Ile-95, and Phe-119) that consistently interact with these pheromones, indicating their critical role in ligand binding. Computational alanine scanning further demonstrated that all five residues act as binding determinants, with Phe-13 and Ile-95 making particularly significant contributions to ligand affinity. The results were further validated by site-directed mutagenesis and fluorescence binding assays. This work provides insights into the function and binding mechanisms of PBPs in sex pheromone recognition and supports the development of targeted mating disruption strategies for P. saucia control. Full article

Ruminants can use volatile fatty acids from rumen fermentation for energy, but substantial starch may bypass the rumen and enter into the small intestine under a high-grain diet. In theory, intestinal starch digestion is energetically more efficient than ruminal fermentation. However, ruminants have inherent limits in starch hydrolysis and glucose transport. Small intestinal starch digestion relies on pancreatic α-amylase. Several studies have indicated that functional amino acids (Leu or Phe) may enhance amylase secretion or activity to improve starch digestion. In contrast, strategies to increase glucose absorption efficiency in the small intestine have received less attention. Thus, this review focuses on the effects of diet, ontogeny, environment, and intestinal microbiota on intestinal glucose absorption and their potential mechanisms. The T1R2/T1R3 glucose-sensing pathways, transporting pathways, and related hormones within the small intestine were systematically reviewed. The advantages and limitations of major approaches regarding glucose absorption including portal vein intubation, nutrient perfusion, everted intestinal sacs in vitro, Ussing chamber, brush-border membrane vesicle, D-xylose test, organoid, and nanosensing are also discussed. Importantly, we propose potential strategies to improve small intestinal glucose absorption (e.g., artificial sweeteners and glucagon-like peptide 2-related modulation). Overall, this review summarizes promising regulatory targets to enhance small intestinal glucose absorption and improve energy efficiency in ruminants. Full article

Ophiocordyceps sinensis, a fungus parasitic on insects, attracts interest due to its unique medicinal applications and complex ecological interactions. The potential relationship between codon usage bias and the parasitic relationship between O. sinensis and Thitarodes xiaojinensis was explored by analyzing the nuclear genomes and mitochondrial genomes of the two. The nuclear genomes showed contrasting preferences: O. sinensis sclerotium exhibited strong GC-ending codon bias (GC3 = 66.13%), while T. xiaojinensis favored AT-ending codons (GC3 = 49.14%). Conversely, both mitochondrial genomes displayed a strong preference (>70% AT) for AT-ending codons. Optimal codon analysis revealed 25 (GC-ending) in O. sinensis and 28 (GC-ending) in T. xiaojinensis nuclear genomes, with overlaps for Leu, Val, Ser, and Pro. The mitochondrial genomes had fewer optimal codons (16 and 13, AT-ending), showing limited overlap (Phe, Leu, Asp, Ser). Neutral plotting analysis, effective number of codons analysis, relative synonymous codon usage analysis, and parity bias analysis showed that the codon usage preferences of the nuclear genomes and mitochondrial genomes of O. sinensis and T. xiaojinensis are jointly affected by selection and mutation pressure. Natural selection predominates in nuclear genomes, while mutation pressure dominates in mitochondrial genomes. This provides new molecular insights into their host–parasite specificity. Full article

Background/Objectives: Staphylococcus aureus virulence is tightly regulated by the agr (accessory gene regulator) quorum-sensing system. Targeting AgrC, the histidine kinase receptor that serves as a core regulator of agr signaling, represents a promising antivirulence strategy that circumvents conventional bactericidal pressure. Methods: In this study, structure-based virtual screening using AutoDock Vina was performed, followed by molecular dynamics simulations, to identify potent analogs of known AgrC inhibitors. Results: A cyclo[Ala-Phe-OLeu-Phe-D-Leu] exhibiting high binding affinity and stable receptor interaction was selected for further evaluation. Antimicrobial susceptibility testing confirmed that the compound did not inhibit bacterial growth. However, at a concentration of 16 µg/mL, it significantly inhibited hemolytic activity with high reproducibility, and the inhibition rate reached 77.60%. Quantitative reverse transcription PCR (RT-qPCR) demonstrated that the compound decreased some key AgrC-mediated genes, including agrC, agrA, saeS, hla, spa, fnbA, and lukS. Conclusions: These findings identify a promising cyclic pentapeptide inhibitor of AgrC that effectively attenuates S. aureus virulence without exerting bactericidal pressure. This work provides a valuable lead compound and offers novel insights for the development of advanced, safe, and effective antivirulence therapeutics. Full article

Copper and zinc nanoparticles have been suggested as potent anticancer agents, particularly against osteosarcoma, a highly aggressive bone cancer with limited treatment options. In order to avoid systemic toxicity, biomolecular carriers able to chelate metal ions and deliver them in a targeted manner to the vicinity of cancer cells need to be developed. Herein, we have used a histidine-containing, cyclic dipeptide as a carrier able to chelate stabilized copper and zinc nanoparticles. The cyclic peptide cyclo-(histidine-phenylalanine) (cHF) self-assembled into amyloid-type fibrils; morphological and structural characterization following metal addition confirmed the formation of cHF−CuNPs and cHF–ZnNPs. These composite nanoparticles demonstrated bacteriostatic activity against Escherichia coli and Staphylococcus aureus at the in vitro level. We evaluated the optimal concentration of cHF–metalNP complexes with limited cytotoxicity to L929 fibroblasts and high cytotoxic effects against MG-63 osteosarcoma cells. Their cytotoxicity was particularly pronounced at pH 6.4, which emulates the tumor microenvironment. The cHF peptide alone did not demonstrate significant antimicrobial or cytotoxic effects to both cell types, suggesting that it can act as a cytocompatible, pH-responsive carrier of metal ions with targeted dual functionality against both microbial infections and osteosarcoma cancer cells. Full article

Background/Objectives: In Phenylketonuria (PKU), engaging in regular physical activity and exercise (PA/E) is important for physical and psychological health, but additional considerations may be required to facilitate uptake and performance as well as to optimise metabolic control. The aim of this study, therefore, was to investigate the stakeholder perspectives on the barriers, facilitators, and solutions to completing PA/E, sport, and nutrition in PKU. Methods: In total, 7 in-person and 6 online semi-structured focus groups (FGs) were conducted with individuals with PKU (n = 31), caregivers (n = 13), clinicians (n = 17), and medical industry professionals (n = 14) in PKU (n = 75 total participants). Three main questions about the barriers, facilitators, and solutions to performing PA/E with PKU were explored. Identified themes were mapped onto the capability, opportunity, motivation, and behaviour (COM-B) model of behaviour change with anonymous quotes from relevant stakeholders used to illustrate the findings. Results: Five common themes were identified. Most notably, individuals with PKU and their caregivers stated fatigue, poor recovery, low energy, and fear around the impact of exercise on blood phenylalanine (Phe) control were barriers to PA/E. Individuals with PKU were aware of the potential benefits of exercise, stating PA/E impacted positively on their mental well-being, daily functioning, and happiness and improved their self-confidence and long-term health. Identified solutions to PA/E participation included greater knowledge in regard to the impact of PA/E on Phe levels, improvements in advice on amount and supplementation with protein substitutes, tailored PKU nutritional advice, more awareness of PA/E within and outside the PKU community, specific PKU guidelines for PA/E, more scientific research, and PA/E events. Misalignment was evident, such that individuals with PKU reported additional barriers to PA/E, whereas other key stakeholder groups perceived the same barriers as the general public. Conclusions: There seems to be a misalignment between individuals with PKU, caregivers, clinicians, and industry professionals regarding PA/E, sport, and nutrition. Individuals with PKU and caregivers reported additional barriers to undertaking PA/E, sport, and nutrition compared to the general public. This suggests that further education and collaboration is needed through stakeholders to better understand how such barriers could be overcome in respect of PA/E, sport, and nutrition in individuals with PKU. Full article

Pathogenic variants in the thyroid-stimulating hormone receptor gene (TSHR) contribute to a wide spectrum of thyroid dysfunctions, ranging from congenital hypothyroidism to thyrotropin resistance. With the advancement of bioinformatics algorithms for variant effect prediction, assessing the pathogenic potential of variants has become increasingly important. This study aimed to investigate the pathogenic effects of TSHR variants classified as variants of uncertain significance (VUSs) in the gnomAD v4.1.0 database. TSHR variants listed in gnomAD v4.1.0 were retrieved and filtered to select missense VUSs based on ClinVar classifications. Multiple bioinformatics tools were used to assess the secondary and three-dimensional structures of the TSHR, as well as protein stability, evolutionary conservation, and molecular dynamics simulations. A total of 2760 TSHR variants were found in gnomAD v4.1.0, including 75 frameshifts, 80 splice-sites, 265 in the 3′ and 5′ untranslated regions, 422 synonymous, 892 others, and 1026 missense variants. Among these, 68 missense VUSs were identified and selected for bioinformatics analysis. Three variants (p.Cys29Trp, p.Leu57Pro, and p.Phe97Ser) were consistently predicted to be pathogenic by all the bioinformatics tools used. All three variants were located within the leucin-rich repeat domain extracellular region of the TSHR and within a highly conserved region across species. Molecular dynamics simulations for mutant proteins (p.Cys29Trp, p.Leu57Pro, and p.Phe97Ser) reveal structural instability in comparison to the wild protein. Comprehensive bioinformatics analysis revealed that three TSHR missense VUSs exhibited pathogenic potential. These variants may contribute to thyroid dysfunction by affecting the receptor’s structural and signalling integrity. Full article

Src family tyrosine kinases regulate oocyte maturation and fertilization in many species, yet their physiological roles in Xenopus laevis (X. laevis) remain incompletely defined. Here, we generated three X. laevis Src (xSrc) constructs with defined point mutations allowing for selective immunochemical detection and controlled modulation of kinase activity: wild type (xSrcWT, Arg121His), constitutively active (xSrcKA, Arg121His/Tyr526Phe), and kinase-negative (xSrcKN, Arg121His/Lys294Met). Capped mRNAs were microinjected into immature oocytes, and effects on meiotic maturation and egg activation were analyzed. All constructs produced detectable Src protein within 4–5 h after injection without inducing progesterone-independent maturation. Following progesterone treatment, MAP kinase phosphorylation, CDK1 activation, and germinal vesicle breakdown (GVBD) occurred normally in all groups, although xSrcKA-expressing oocytes showed a modest but reproducible acceleration of MAPK activation and GVBD. Global tyrosine phosphorylation analysis revealed increased phosphorylation of several proteins, including a prominent ~50 kDa substrate, specifically in xSrcKA oocytes. After maturation, oocytes were subjected to artificial activation. xSrcKN-expressing oocytes responded normally to Ca²⁺ ionophore (A23187), indicating that Src activity is not required for direct Ca²⁺-mediated activation. In contrast, xSrcKN oocytes exhibited markedly reduced activation in response to hydrogen peroxide or Cathepsin B, which stimulate membrane-associated signaling pathways. These findings demonstrate that Src kinase activity is required for membrane signal-mediated egg activation but is dispensable for activation driven by direct intracellular Ca²⁺ elevation. Collectively, our results identify Src kinase as a positive regulator of progesterone-induced meiotic maturation and a critical mediator of specific fertilization-like activation pathways in X. laevis. Full article

Ocimum (basil) is a globally significant medicinal and culinary herb. While its bioactive secondary metabolites are well-studied, the medicinal potential of its abundant primary metabolites (amino acids, vitamins, carbohydrates, steroids) remains largely unexplored. To address this gap, we employed an integrated multi-omics strategy. First, UPLC-MS/MS-based metabolomics quantified primary metabolites across six distinct Ocimum accessions (Ocimum × africanum, Ocimum tenuiflorum, Ocimum gratissimum). Profiling identified 291 primary metabolites, revealing significant interspecific variation, with 273 differential accumulated metabolites (DAMs). Subsequent network pharmacology analysis of 61 high-impact DAMs predicted 516 potential targets. Protein–protein interaction refinement yielded 28 core targets, predominantly integrins (ITGB1, ITGB3, ITGA4, ITGA2B, ITGAV) and kinases (IGF1R, PIK3CA, SRC). Enrichment analysis implicated these targets in focal adhesion, ECM-receptor interaction, and PI3K-Akt signaling pathways. Molecular docking confirmed strong potential binding (binding energy < −7 kcal/mol) between key tripeptides (e.g., Met-Ser-Tyr, Phe-Cys-Gln) and integrin subunits. Antioxidant assays (DPPH, ABTS, FRAP) further showed significant genotypic variation. This study systematically deciphers the primary metabolome of Ocimum and, through a multi-omics approach, reveals novel integrin-mediated mechanisms underpinning its potential therapeutic value, providing a foundation for developing basil-based nutraceuticals and pharmaceuticals. Full article

Walnut peptide Lys-Gly-His-Leu-Phe-Pro-Asn (KG-7) is a food-derived bioactive peptide with a high antioxidant capacity. We systematically evaluated the ameliorative effects of KG-7 on scopolamine-induced memory deficits in mice and its intestinal absorption mechanisms through integrating motion behavior analysis, molecular biochemistry research, and fluorescence imaging technology. Morris water maze tests revealed that KG-7 significantly improved the behavioral performance of these mice. Further mechanistic investigations demonstrated that KG-7 restored cholinergic function by reducing acetylcholinesterase activity and increasing acetylcholine levels. Hematoxylin-eosin staining and hippocampal immunohistochemistry confirmed that KG-7 alleviated neuronal damage by downregulating Hes1 overexpression, clarifying its behavioral improvement mechanism. In vitro fluorescence imaging showed that KG-7 reached peak accumulation in brain tissue 8 h post-administration, confirming its brain delivery. To elucidate the absorption mechanism, immunohistochemistry and immunofluorescence revealed that KG-7 markedly reduced the expression of efflux transporter P-gp in the small intestine, thereby diminishing efflux activity, while weakened tight junction (Occludin, ZO-1) fluorescence indicated activation of the paracellular pathway. Western blot analysis confirmed that KG-7 enhanced paracellular absorption efficiency and reduced intestinal efflux by downregulating ZO-1, Occludin, and efflux transporters (P-gp, BCRP, and LRP1) alongside upregulating Claudin-2 expression. These findings provide a foundation for exploring walnut peptides that enhance memory and optimize absorption. Full article

Background/Objectives: Casein glycomacropeptide (cGMP) has been modified to enable its suitability as a low phenylalanine (Phe) protein substitute (PS) in phenylketonuria (PKU). No data is available about its global usage. Methods: A 60-item multiple choice and short answer/extended response questionnaire examining the use of modified cGMP in PKU was distributed globally to dietitians and physicians via web-based professional inherited metabolic disorder groups. Results: Respondents (n = 208) from 45 countries across 6 continents completed the questionnaire. Of these, 83.7% (n = 174) were dietitians/nutritionists, 14.9% (n = 31) medical doctors/physicians and 1.4% (n = 3) other health professionals, caring for both paediatric and adult patients (59.1%), paediatrics only (25.0%) or adults only (15.9%). cGMP PS were reported as not available in their centre/hospital by 19.7% (n = 41), mostly in Africa, South America, and southern and western Asia. The main reasons included lack of regulatory approval (65.8%), not promoted by manufacturers (41.5%), and cost (29.3%). An estimated 25% of represented patients globally were using cGMP PS; 78.4% (n = 163) following refusal/poor adherence with Phe-free amino acids and 54.8% (n = 114) for adult patients recommencing dietary treatment. There were concerns about the residual Phe in cGMP negatively impacting blood Phe levels in children <12y (66.3%), adolescents (48.0%), adults (34.6%), and the first trimester of pregnancy (53.1%). Sixty nine percent (n = 145) adjusted dietary Phe prescription according to the cGMP Phe content, particularly in regions with a higher percentage of severe PKU variants. Commonly perceived clinical advantages with cGMP were improved taste/palatability (93.2%, n = 194) and fewer gastrointestinal symptoms (55.8%, n = 116). Perceived clinical disadvantages were residual Phe (72.1%, n = 150), lack of data in children < 3 years (48.1%, n = 100), and the high energy content of some brands (45.2%, n = 94). There were concerns that cGMP PS were too high in sugar (34.1%, n = 71) and dissatisfaction or uncertainty about the adequacy of its Phe (66.3%) and amino acid (34.1%) content. Conclusions: There is global inconsistency in access to cGMP PS suitable for PKU, and in the interpretation of evidence-based research. Some professionals have significant concerns about its nutritional composition particularly residual Phe, limiting its estimated use to approximately 25% of PKU patients globally. Full article

Background/Objectives: Sodium-glucose co-transporter 2 (SGLT2) inhibitors are essential antidiabetic medications. However, their side effects warrant careful consideration. The search for novel SGLT2 inhibitors with high affinity remains an ongoing endeavor. Medicine food homology (MFH) herbs show promise for drug development due to their nutritional and medicinal value. Methods: This study aims to address the shortcomings of existing virtual screening models for SGLT2 inhibitors by optimizing feature selection and integrating multidimensional molecular fingerprints. Subsequently, an integrated virtual screening pipeline is constructed to identify potential SGLT2 inhibitors from eight selected MFH herbs. Results: The results indicate that the optimal model (LightGBM and RF) achieved an accuracy of 0.97 and an AUC of 0.98. Following rigorous filtering, a total of 44 potential SGLT2 inhibitors were identified, among which, Isoononin (from Gancao) and Ononin (from Huangqi, Gegen, and Gancao) exhibit favorable drug likeness and safety. Molecular docking demonstrate that both compounds can effectively bind to the SGLT2 active site, establishing stable hydrophobic interactions with critical residues such as Phe98 and Phe453. Furthermore, molecular dynamics simulations confirm the stability of the interactions between the two compounds and SGLT2. Conclusions: This study significantly enhances the accuracy and stability of SGLT2 inhibitor virtual screening models by addressing deficiencies in structural characterization and feature selection. It provides candidate molecules for the development of novel SGLT2 inhibitors and offers new scientific evidence for the application of MFH herbs in the prevention and treatment of chronic metabolic diseases. Full article