Search Results (2,878)

Background: Foetal structural abnormalities can be detected in approximately 3% of all pregnancies and frequently remain without a genetic diagnosis. This study aims to apply an integrated approach with the final goal of providing a molecular diagnosis in the challenging Italian setting of early termination of pregnancy. Methods: In a cohort of 86 foetuses, post-mortem dysmorphological examination, radiological assessments, and molecular autopsy through Whole-Exome Sequencing—WES—analysis were performed. Results: Forty-two foetuses were phenotypically classified as presenting a single major malformation (i.e., central nervous system, skeletal, urogenital, or cardiac anomalies, or fluid accumulation), while 44 foetuses presented multiple malformations and/or dysmorphic features. Overall, WES provided a diagnostic yield of 26.7%; additionally, seven Variants of Uncertain Significance (VUS) potentially liked to the foetal phenotype were identified. The highest detection rate was achieved for foetuses presenting a single major urogenital (50%) or skeletal (42.9%) malformation, followed by foetuses presenting multiple malformations (27.3%). Peculiar results of particular interest were (1) the identification of two splicing variants (within the INPPL1 and RHOA genes), functionally characterised through minigene assay, which contributed to evaluate their pathogenicity, and (2) the identification of a novel de novo missense ZNF292 variant (NM_015021.3:c.6325A>C p.(Ser2109Arg)) in a foetus affected by corpus callosum hypoplasia. The ZNF292 gene is associated with the Intellectual developmental disorder, autosomal dominant 64 and this finding represents the first report of prenatally detected anomalies of the central nervous system in a foetus carrying a ZNF292 variant. Conclusions: This study underlines the diagnostic utility of an integrated approach to achieve a precise genetic diagnosis for structural foetal abnormalities, thus providing families with precise recurrence risk estimations and detailed options about future pregnancies. Additionally, a systematic implementation of this strategy could be crucial to better characterise new variants and discover new genes involved in embryonic and foetal development. Full article

The global rise of antimicrobial resistance (AMR), exacerbated by the COVID-19 pandemic, has led to a surge in infections caused by multidrug-resistant (MDR) bacteria. A key driver of this phenomenon is co-selection, where exposure to one antimicrobial promotes resistance to others via horizontal gene transfer (HGT) mediated by mobile genetic elements (MGEs). Carbapenem-resistant Enterobacteriaceae, known for their genomic plasticity, are particularly worrisome; yet genomic data from Latin America—especially Ecuador—remain scarce. This study investigated four carbapenem-resistant clinical isolates (two Klebsiella pneumoniae ST1440 and two Serratia marcescens) from tracheal aspirates of three ICU patients during a COVID-19 outbreak at Hospital IESS Quito Sur, Ecuador. Phenotypic profiling and whole-genome sequencing were performed, followed by bioinformatic reconstruction of plasmid content. Nineteen plasmids were identified, carrying 70 resistance-related genes, including antimicrobial resistance genes (ARGs), metal resistance genes (MRGs), integrons, transposons, and insertion sequences. Hierarchical clustering revealed six distinct gene clusters, with several co-localizing ARGs and genes for resistance to disinfectants and heavy metals—suggesting strong co-selective pressure. Conjugative plasmids harboring high-risk elements such as blaKPC-2, qacE, and Tn4401 were found in multiple isolates, indicating potential interspecies dissemination. These findings emphasize the importance of plasmid-mediated resistance during the pandemic and highlight the urgent need to enhance genomic surveillance and infection control, particularly in resource-limited healthcare settings. Full article

Ozone (O₃) therapy has demonstrated antioxidant and anti-inflammatory properties, but the systemic administration of ozonated saline solution (O₃SS) remains underexplored. This study evaluates the cytotoxicity, antioxidant response, and immunomodulatory effects of O₃SS on murine microglial (BV2) and human endothelial (HUVEC) cells. Cells were exposed to increasing doses of O₃ (1, 5, or 10 μg/NmL) dissolved in saline. Viability assays showed that low doses (1 and 5 μg/NmL) enhanced cell proliferation without cytotoxicity, while the highest dose (10 μg/NmL) reduced viability and increased cell death. O₃SS treatment upregulated antioxidant genes, including Nrf2 and SOD1, and decreased reactive oxygen species in lipopolysaccharide (LPS)-stimulated microglia. Additionally, O₃SS modulated microglial phenotype by reducing pro-inflammatory markers (iNOS, IL-1β) and increasing anti-inflammatory markers (Arg-1, IL-10). Immunofluorescence confirmed enhanced Arg-1 protein expression, indicating a shift toward an anti-inflammatory state. These results suggest that low-dose O₃SS activates cellular antioxidant defenses and promotes an anti-inflammatory microglial phenotype, supporting its potential as a safe systemic O₃ therapy. Further studies are warranted to confirm in vivo efficacy and optimize clinical protocols. Full article

Non-human primates (NHPs) are close relatives of humans and can serve as hosts for many zoonotic pathogens. They play crucial role in spreading antimicrobial resistant bacteria (AMR) to humans across various ecological niches. The spread of antimicrobial resistance in NHPs may complicate wildlife conservation efforts, as it may threaten domestic livestock, endangered species as well as human’s health. This review analyses the existing literature on the prevalence of AMR in NHP species, including Rhinopithecus roxellana, Macaca fascicularis, and Sapajus nigritus, to create awareness in all stake holders involve in the fight against AMR on the serious potential threats that these primates pose. Methods: We performed a comprehensive literature search using the PubMed (National Library of Medicine-NLM), Scopus (Elsevier), Web of Science Core Collection (Clarivate Analytics), Springer Link (Springer), and Science Direct (Elsevier) databases until January, 2025. The search strategy combined terms from the areas of non-human primates, antibiotic resistance, antimicrobial resistance, and antibacterial resistance genes (ARGs). Studies that isolated bacteria from NHPs and assessed phenotypic resistance to specific antibiotics as well as studies that identified ARGs in bacteria isolated from NHPs were included. Data were synthesised thematically across all included studies. Results: A total of 37 studies were included (explained as Cercopithecidae (n = 23), Callithrix (n = 6), Cebidae (n = 4), Hominidae (n = 3), and Atelidae (n = 1)). The results showed that the most common ARB across the various NHPs and geographical settings was Staphylococcus spp. (45.95%) and Escherichia spp. (29.73%). The tested antibiotics that showed high levels of resistance in NHPs included Tetracycline (40.54%), Ciprofloxacin (32.43%), and Erythromycin (24.34%), whereas ermC, tetA, tetM, aadA, aph (3″)-II, and qnrS1 were the most widely distributed antibiotic resistance genes in the studies. Conclusion: NHPs are potential natural reservoirs of AMR, therefore global policy makers should consider making NHPs an indicator species for monitoring the spread of ARB. Full article

Background: Dilated cardiomyopathy (DCM) and long QT syndrome (LQTS) are genetically heterogeneous cardiac disorders that contribute significantly to morbidity and sudden cardiac death. Although they are typically considered distinct entities, co-occurrence within families has been increasingly recognized, complicating diagnosis and genetic counseling. Identifying shared genetic determinants may provide insights into overlapping disease mechanisms. Methods: We investigated a multi-generational family in which several members presented with features of both DCM and LQTS. Exome sequencing was performed to identify potential disease-causing variants, and candidate findings were validated by Sanger sequencing. In silico prediction tools and evolutionary conservation analysis were used to assess the pathogenic potential of the identified variant. Results: We identified a novel heterozygous missense variant in the CRYAB gene, c.368G>A (p.Arg123Gln). This variant is located in a highly conserved region critical for protein function and was consistently predicted to be deleterious across multiple computational algorithms. Segregation analysis demonstrated co-occurrence of the variant with disease phenotypes in affected family members. Clinically, several carriers exhibited overlapping features of both DCM and prolonged QT interval, suggesting a dual cardiac phenotype associated with this mutation. Conclusions: Our findings expand the phenotypic spectrum associated with CRYAB mutations, linking them to a combined presentation of dilated cardiomyopathy and long QT syndrome. This underscores the importance of including CRYAB in comprehensive gene panels for inherited cardiac disorders and highlights the need for integrated clinical and genetic evaluation in families presenting with complex cardiac phenotypes. Full article

Polyamines, particularly spermidine, have emerged as key dietary factors with roles in cellular health, autophagy, and longevity. However, strategies for scalable production of polyamine-rich food ingredients remain limited. Here, we report the development of a high-spermidine-producing Saccharomyces cerevisiae strain, 3L63, obtained via ultraviolet mutagenesis of the K7 strain. This strain exhibited a 5.9-fold increase in the total polyamine content, with spermidine being the most abundant. A scalable fermentation system of up to 10⁴ L was established, yielding a dried yeast product that met food safety criteria. Whole-genome sequencing identified mutations in central metabolic pathways, including ARG3, and functional enrichment analysis suggested broad metabolic rewiring, supporting an enhanced biosynthetic capacity, including polyamines. Free amino acid profiling revealed higher arginine levels in 3L63 than in K7, which is consistent with its role as a polyamine precursor. The 3L63 yeast-derived product was enriched in essential amino acids and polyamines. Functionally, this strain promoted the proliferation of normal and senescent human dermal fibroblasts, and its autophagy-inducing activity exceeded that of equivalent concentrations of pure spermidine, suggesting synergistic effects of yeast-derived bioactive compounds. This study demonstrates a non-genetically modified, high-spermidine yeast strain as a promising functional food ingredient with potential applications in healthy aging. Full article

HaWRKY33 is induced by salicylic acid and participates in the disease resistance signaling pathway of sunflower rust disease; however, the transcriptional regulatory mechanism of this protein against Sclerotinia sclerotiorum in sunflowers remains unclear. Given this, we conducted a survey of 426 sunflower accessions at the natural disease nursery in Gannan County and identified a single dominant physiological race, MCG1, using simple sequence repeat methods. Additionally, we performed indoor inoculation tests using this dominant race and obtained disease-resistant varieties, W227 and BC2202-03, as well as susceptible varieties, N241 and Z155. Further, we inoculated the above resistant and susceptible combination materials with MCG1 and conducted transcriptomic analysis and RT-qPCR validation. Through KEGG analysis, we found that HaWRKY33 is involved in the plant–pathogen interaction pathway, suggesting that HaWRKY33 may regulate sunflower defense responses against Sclerotinia sclerotiorum through the plant–pathogen interaction pathway. Finally, yeast two-hybrid screening and AI prediction using AlphaFold 3 revealed strong interactions between ARG-189 and GLU-344 amino acids in the HaWRKY33-AOA251SVV7 proteins, indicating that the HaWRKY33-AOA251SVV7 pattern regulates the sunflower defense response against Sclerotinia sclerotiorum in a transcriptional complex form. In summary, these results provide new insights into the disease resistance mechanisms of sunflowers against Sclerotinia sclerotiorum and promote the development of molecular breeding for sunflower resistance to Sclerotinia sclerotiorum. Full article

Swine manure poses significant challenges for anaerobic digestion due to its low carbon-to-nitrogen (C/N) ratio and elevated ammonia concentrations, both of which restrict methane generation. This study investigated the impact of integrating low-intensity ultrasound with co-digestion of piggery wastewater and food waste leachate. Laboratory-scale upflow anaerobic sludge blanket (UASB) reactors were employed under four operational conditions to evaluate anaerobic digestion performance, track shifts in microbial community structure, and assess the abundance of antibiotic resistance genes (ARGs). Co-digestion significantly enhanced methane production, yielding 1.3–3.2 times more than manure alone, while low-intensity ultrasound further increased methane yields by approximately 36–44% at high loading rates. Moreover, coupling low-intensity ultrasound with co-digestion led to the most rapid recovery following an overloading shock. Unexpectedly, ultrasound treatment alone increased the expression of certain ARGs (tetG, sul1, ermB) and the Integrase gene (intI1), while co-digestion led to a reduction in these genetic markers. These findings clearly indicate that the concurrent application of co-digestion and low-intensity ultrasound achieved the highest methane yield, the fastest recovery after organic overloading, and greater suppression of specific ARGs. Full article

Bothrops species are responsible for the majority of envenomations in Argentina. In particular, Bothrops diporus is among the main species responsible for the majority of envenomations in Argentina and causes significant injury and coagulopathy. Given the significance of this venom, the authors sought to define the toxin responsible for coagulopathy with specialized spectrophotometric and thromboelastographic methods. Utilizing clotting time, spectrophotometry, and thromboelastography, it was determined that B. diporus venom has potent, procoagulant activity in human plasma and buffer milieu. Calcium-dependent and -independent activities consistent with serine protease activity were identified. The activity included both thrombin-generating and thrombin-like enzymatic activity. The venom cleaved the serine protease-specific chromogenic substrate β-Ala-Gly-Arg-p-nitroanilide diacetate, and its activity was inhibited in plasma by antithrombin after addition of heparin. Further, venom exposed in isolation to RuCl₃, a known inhibitor of serine protease-containing venoms, demonstrated decreased activity in human plasma. In conclusion, the present study contributes to a better understanding of B. diporus venom and may have implications for the rational design of inhibitors, antivenom formulations, or preclinical models to study venom-induced coagulopathies. Full article

Despite significant advances in understanding the genetics of Parkinson’s disease (PD) and Parkinsonism, the diagnostic yield remains low. Pathogenic variants of GBA1, which encodes the lysosomal enzyme β-glucocerebrosidase and causes recessive Gaucher dis-ease, are recognized as the most important genetic risk factor for PD in heterozygous carriers. This study focuses on the functional genomics of rare genetic variations in other lysosomal hydrolytic enzymes genes in patient-derived fibroblasts. We examined 49 early-onset PD patients using whole exome sequencing and in silico panel analysis based on a curated PD gene list. Two patients were found to carry the p.Asp313Tyr variant in the X-linked GLA gene (encoding GALA, typically associated with Fabry disease), and one patient carried the p.Arg419Gln variant in GLB1 (encoding β-Gal, linked to the recessive GM1 gangliosidosis and mucopolysaccharidosis type IVB). The in silico study of both variants supports a potentially damaging impact on the encoded protein function and structural destabilization. Additional candidate variants were found related to lysosomes, Golgi apparatus and neurodegeneration, suggesting a multifactorial contribution to the disease. However, none of these variants met diagnostic standards. Functional assays showed a significant decrease in GALA expression and partial retention of the enzyme in the trans-Golgi network in fibroblasts with GLA:p.Asp313Tyr, while altered Golgi morphology was observed in fibroblasts with GLB1:p.Arg419Gln. Moreover, all patients exhibited abnormalities in lysosomal morphology, altered lysosomal pH, and impaired autophagic flux. Our findings suggest that rare, heterozygous variants in lysosomal-related genes, even when individually insufficient for monogenic disease, can converge to impair lysosomal homeostasis and autophagic flux in EOPD. The underlying genetic and cellular heterogeneity among patients emphasizes the importance of combining genetic and functional approaches to better understand the mechanisms behind the EOPD, which could enhance both diagnosis and future treatments. Full article

The VRTN (vertebrin) gene plays a crucial role in regulating thoracic vertebral number in mammals; however, its function in goats remains largely unexplored. This study aimed to investigate genetic variations in the VRTN gene in Hainan black goats (HNBGs) and evaluate their associations with growth and meat traits. Using whole-genome and Sanger sequencing, we identified four SNPs in the VRTN gene, including three missense mutations (p.Pro615Ser, p.Arg490Lys, p.Thr476Met) and one synonymous mutation (p.Asp688Asp). Tissue expression profiling revealed high VRTN mRNA expression in skeletal muscle and low expression in internal organs, suggesting a potential role in muscle development. Temporal expression analysis indicated dynamic regulation during growth, with higher expression levels observed in early developmental stages. Association analyses revealed significant correlations between specific SNPs and key traits, including body length, chest circumference, carcass weight, and meat quality parameters. Notably, the p.Pro615Ser mutation was associated with a 0.441-fold reduction in VRTN expression and showed strong associations with multiple traits, underscoring its functional importance. These findings demonstrate that VRTN polymorphisms influence growth and muscle development in HNBGs, providing valuable insights for marker-assisted selection in goat breeding. Full article

The secondary contamination of nodularin disinfection by-products (NOD-DBPs) is a problem worthy of attention. In this study, prototypical NOD-R-DBPs were prepared, and their toxicity was assessed using conventional protein phosphatase (PPs) inhibition assay, confirming that structural changes in “Adda³” during chlorination are key factors leading to a significant reduction in NOD-R toxicity. However, some NOD-R-DBPs still exhibit certain levels of toxicity (2.8–81% of NOD-R). To elucidate the mechanism underlying the potential inhibitory effect of NOD-R-DBPs on protein phosphatase 2A (PP2A), molecular simulations were employed to establish interaction models between prototypical NOD-R-DBPs and PP2A using homology modeling strategies, and molecular docking was used to obtain candidate interaction parameters between prototypical NOD-R-DBPs and PP2A. Structural changes in “Adda³” weakened the hydrogen bonds “Adda³”Asn₁₁₇ and “Adda³”His₁₁₈. Subsequently, the disruption of “Adda³” altered key interactions between NOD-R-DBPs and PP2A (hydrogen bond Mdhb⁵ ← Arg₈₉, ionic bond Glu⁴-Arg₈₉, metal bond His₂₄₁-Mn₁²⁺, etc.). The changes in these interactions further altered the interactions between conserved amino acids and the catalytic center Mn²⁺ (ionic bond Asp₅₇-Mn₂²⁺), thereby increasing Mn²⁺ exposure. Meanwhile, the retained interactions promoted the binding of -PO₄ with the conserved amino acids His₁₁₈ and Arg₈₉. Prototypical NOD-R-DBPs retained the aforementioned key interactions and thus exhibit potential inhibitory effects on PP2A. The varying degrees of damage to the Adda³ structure led to significant differences in the inhibitory effects of different NOD-R-DBPs on PP2A. Full article

Agricultural pesticides are significant drivers of antibiotic resistance in soil. However, the differential impacts of antibiotic versus non-antibiotic pesticides on the soil resistome are poorly characterized. Here, we analyzed sequencing data from soils exposed to either antibiotic or non-antibiotic pesticides to compare differences in antibiotic resistance gene (ARG) burden, diversity, assembly processes, network topology, and host taxonomy. Soils exposed to antibiotic pesticides exhibited a significantly higher ARG burden (0.52% vs. 0.27% of total genes), whereas soils exposed to non-antibiotic pesticides showed significantly higher alpha diversity (p < 0.05). ARG community compositions also differed significantly between antibiotic and non-antibiotic exposures (PERMANOVA, R² = 0.215, p < 0.001). Assembly analysis using the modified stochasticity ratio indicated that deterministic processes governed ARG community assembly in both groups, with stronger influence observed in non-antibiotic soils. Co-occurrence network analysis revealed contrasting patterns. A compact, highly centralized network emerged in antibiotic-exposed soils, while a larger, more dispersed network characterized non-antibiotic soils. In both networks, aminoglycoside ARGs served as keystone nodes, accompanied by the β-lactam ARG in antibiotic soils and the macrolide ARG in non-antibiotic soils. Pseudomonadota was the predominant ARG host (>60% contribution) across both exposures, though many other phyla exhibited significance (p < 0.05) between group differences in their ARG contributions. Non-pathogenic bacteria comprised the majority of ARG hosts in all samples. When examining ARG contributions from pathogenic hosts, zoonotic and animal-associated pathogens contributed significantly (p < 0.01) more in non-antibiotic soils than in antibiotic soils, whereas the ARG contribution from plant pathogens was comparable between the two pesticide groups. Overall, our study suggests that antibiotic and non-antibiotic pesticides shape distinct ARG network patterns and host–pathogen profiles, posing distinct risks to public health and agricultural ecosystems. Full article

Background: Type I collagen is the most abundant protein of the extracellular matrix. Pathogenic variants in COL1A1 or COL1A2 are classically associated with osteogenesis imperfecta (OI) and Ehlers–Danlos syndrome (EDS). An emerging clinical entity—COL1-related overlap disorder—encompasses individuals exhibiting phenotypic features of both conditions. Methods: We report a 55-year-old male presenting with disproportionate short stature, grayish-blue sclerae, multiple fractures, long bone deformities, joint hypermobility, and atrophic surgical scarring. The patient also had long-standing, untreated childhood-onset hypopituitarism. Imaging studies revealed numerous prior fractures, bowing of forearm bones, and multiple Wormian bones. Results: Genetic testing confirmed a novel heterozygous COL1A1 exon 14 variant (c.940G > A, p.Gly314Arg), presenting with a phenotype consistent with a COL1-related overlap syndrome. Conclusions: This case expands the phenotypic spectrum of COL1A1 mutations and supports the concept of COL1-related phenotypic overlap. Full article

Oxidative stress, caused by excessive free radicals, leads to cellular damage and various diseases. Antioxidant peptides from natural proteins offer potential in alleviating this stress. In this study, antioxidant peptides were identified from deer antler proteins using in silico enzymatic hydrolysis and machine learning. Peptides with high prediction scores and non-toxic profiles were selected for evaluation. The antioxidant activities of top candidates, PHPAPTL and VPHGL, were confirmed by radical scavenging assays and their protective effects in HepG2 cells. Molecular dynamics simulations revealed stable binding of these peptides to Keap1, enhancing system stability and reducing residue fluctuations at the ligand-binding interface. Key interactions involved Arg415, Arg483, Arg380, and Ser555. Secondary structure analysis showed peptide binding induced local conformational transitions, notably increasing parallel β-sheet formation near active sites. These findings provide mechanistic insight into their antioxidant effects and support their potential application in functional food development. Full article