Search Results (2,706)

Background/Objectives: Variants in the RYR1 gene are associated mainly with Malignant Hyperthermia. Missense variants are largely the most common, while insertions and duplications account for less than 10%. We aimed to investigate the effect of a rare duplication in the RYR1 gene with the variability of the Malignant Hyperthermia susceptibility phenotype. Methods: We used exome variant screening, in vitro contracture test, anatomopathological examination of the muscle biopsy, RT-qPCR analysis for RYR1 relative expression. Results: We identified a family with two affected siblings carrying an insertion of 18 pair bases in exon 91 of the RYR1 gene, resulting in an in-frame duplication of 6 amino acids (c.12835_12852 dupGAGGGCGCGGCGGGGCTC: 162 p.G4279_T4284insAAGLEG). This variant was found at a frequency of 0.0007% in gnomAD and was absent in 1200 Brazilian controls. First classified as a Variant of Uncertain Significance (VUS), with the molecular and physiological data from our family, we were able to reclassify it, reaching 5 points, which is still a VUS but borderline likely pathogenic. Muscle relative mRNA expression of RYR1 in the two patients identified a ~50% reduction, suggesting a possible hypomorphic allele. Conclusions: The pathomechanisms of RYR1 gene variants in Malignant Hyperthermia are mainly associated with gain-of-function mechanisms, but small insertions can often lead to loss of function or improper folding protein. This study adds evidence to the possibility that duplication in this region can cause structural defects and a more severe phenotype in the patients. Full article

Background: The water-soluble Grifola frondosa polysaccharides (GFPs) are the primary bioactive component of the edible and medicinal fungus Grifola frondosa. However, the digestive behavior of GFPs in the human gastrointestinal (GI) tract and their subsequent interaction with gut microbiota (GM) to exert health effects remain unclear. Methods: In this study, GFPs were extracted based on a traditional hot water decoction. An in vitro simulated GI digestion model and a human fecal microbiota fermentation model were established to systematically investigate the digestive stability of GFPs, GM modulation, and metabolite changes. Results: Results showed that GFPs remained structurally stable during in vitro oral, gastric, and small intestinal digestion, allowing them to reach the colon intact for microbial fermentation. During colonic fermentation, GFPs were efficiently degraded by GM, and significantly increased the relative abundance of beneficial bacteria such as Akkermansia, Bacteroides, Parabacteroides, and Lactobacillus while reducing the abundance of pathogenic Escherichia-Shigella. Meanwhile, GFPs enriched metabolites beneficial for intestinal health, among which γ-aminobutyric acid (GABA) was the most significantly upregulated. Single-strain fermentation confirmed that Lactobacillus (L. plantarum) was the core GABA-producing genus. Conclusions: This study highlights the potential of GFPs as prebiotics for GM modulation, expands the understanding of the health-promoting effects of fungal polysaccharides, and provides a theoretical basis for the development of GFP-based functional foods. Full article

Pasteurella multocida is capable of infecting various animal species, causing hemorrhagic septicemia or infectious pneumonia, and it is widely prevalent and pathogenic worldwide. However, the prevention and control technologies for animal pasteurellosis remain relatively under-developed, with a heavy reliance on the traditional inactivated vaccines and live attenuated vaccines developed in the mid-20th century. A previous systematic review summarized research progress in the field of inactivated vaccines for animal pasteurellosis and revealed that inactivated vaccines exhibit high levels of homologous protective efficacy, albeit with a relatively short duration of immunity and limited cross-protection between different serotypes. Live attenuated vaccines are known to exhibit significant advantages, including a prolonged duration of immunity, strong cross-protective capacity, and low production costs. Throughout the history of vaccine development, various live attenuated vaccines have been extensively used as critical tools for preventing and controlling infectious diseases in animals and humans and combating antimicrobial resistance, substantially reducing the morbidity and mortality rates of regional or international epidemics. The developmental history of live attenuated vaccines against Pasteurella multocida has primarily encompassed three distinct research and application periods, characterized by natural attenuated vaccines, traditional attenuated vaccines, and genetically engineered live attenuated vaccines. Here, we comprehensively review the research and application history of natural attenuated vaccines for Pasteurella multocida. Our aim was to provide theoretical foundations and novel perspectives for the development of a new generation of live attenuated vaccines against pasteurellosis in livestock and poultry. Full article

Branch canker and dieback, caused by Botryosphaeriaceae and Diaporthaceae, is a major disease of walnut (Juglans regia L.) worldwide. In Chile, the impact of pruning wound age and timing on susceptibility to these pathogens in walnut trees remains poorly understood. During June–July (2023) and June–July (2024), this study assessed the effect of pruning wound age of the walnut cv. Chandler on infection by seven fungal species and simultaneously tracked seasonal conidial release of Botryosphaeriaceae spp. in the Maule Region, Chile. Lignified twigs were artificially inoculated at 1, 15, 30, and 45 days after pruning, and necrotic lesion lengths were measured six months post-inoculation. All fungal isolates caused significantly longer lesions than the control (p < 0.0001), with Diplodia mutila, Neofusicoccum nonquaesitum, and N. parvum being the most aggressive. At the same time, Dothiorella sarmentorum and Diaporthe species (Diaporthe australafricana, Di. foeniculina, and Di. patagonica) produced the smallest lesions. Susceptibility decreased with increasing wound age, with a significant interaction between fungal species and pruning wound age. Spore trapping of Botryosphaeriaceae revealed that dispersal was positively associated with rainfall (r = 0.81, p < 0.0001), relative humidity (r = 0.51 to 0.61, p < 0.05) and average temperature (r = 0.32 to 0.58, p < 0.05), but negatively or not significantly correlated with maximum temperature (r = −0.59 to −0.79, p > 0.05). These results demonstrate that rainfall or relative humidity, moderate conditions, and favor conidial release. At the same time, infection risk declines with wound age, underscoring the need to adjust pruning schedules and preventive strategies to reduce disease risk in walnut orchards. Full article

Recently, we observed that pars planitis (PP) patients present alterations in peripheral blood (PB) Th17/Treg associated with dysregulation in the Th1 response. Yet, little is known about the systemic distribution of myeloid cells, which drive the recruitment and differentiation of the adaptive effectors toward pathogenic inflammatory Th1 and Th17 as well as regulatory lymphocytes in PP. Although myeloid populations in patients with uveitis have previously been addressed, the data did not provide an exact description of PP patients. Using flow cytometry, we evaluated monocyte and IDO-expressing monocyte-like myeloid-derived suppressor cell (MDSC) subpopulations in PB samples from 15 patients with different courses of PP (cystoid macular edema and non-macular edema subgroups; CME and nCME, respectively) and 17 healthy controls (HCs) in relation to the Th1, Th17, and immunoregulatory subsets. We observed that only PP patients from the CME subgroup presented a significantly higher fraction of CD16⁺ IDO-expressing MDSCs and intermediate CD14^highCD16⁺ monocytes compared to the HCs; this corresponded with relative up-regulation of Th1 and Th17, and down-regulation of Treg. In addition, alongside the increased percentage of IDO-expressing CD16+ MDSCs, the MDSC compartment displayed an inappropriate level of IDO (more pronounced in the CD16⁻ subset) only in CME patients. At the same time, the fraction of CD16⁻ myeloid cells did not differ significantly among the patient cohorts and healthy participants. Our study is the first to evaluate subpopulations of circulating myeloid cells in PP patients and indicates that an increased fraction of CD16⁺ myeloid cells might reflect the immunological and clinical severity of PP. Full article

Extracellular vesicles (EVs) are bilayer-membrane cellular components that deliver protected cargo to the extracellular environment and can mediate long-distance signaling. We have previously reported that EVs isolated from the virulent fungal pathogen Paracoccidioides brasiliensis Vpb18 can revert the expression, in the attenuated variant Apb18, of stress-related virulence traits. We presently show that the Vev and Aev, respectively, produced by these variants display distinct proteomes, with prevalent functional enrichment in Vev related to oxidative stress response, signal transduction, transport, and localization, in addition to richer protein–protein interaction. Proteome sequences were obtained by nanoflow liquid chromatography coupled with tandem mass spectrometry (nano LC-ESI-MS/MS). The Vev and corresponding Vpb18 proteomes also differed, suggesting a selective bias in vesicle protein cargo. Moreover, sublethal oxidative (VevOxi) and nitrosative (VevNO) stress modulated the Vev proteome and a positive correlation between VevOxi/VevNO-enriched and Vev-enriched (relative to Aev) proteins was observed. Out of 145 fungal virulence factors detected in Vev, 64% were enriched, strongly suggesting that molecules with virulence roles in Paracoccidioides are selectively concentrated in Vev. Our study significantly advanced the field by exploring protein N-terminal acetylation to a dimension rarely investigated in fungal EV proteomics. The proportion of N-terminally acetylated proteins in Vev was higher than in Vpb18 and the presence of Nt-acetylation in Vev-enriched virulence factors varied across the samples, suggesting that it might interfere with protein sorting into EVs and/or protein functionality. Our findings highlight the relevance of our fungal model to unraveling the significance of fungal EVs in pathogenesis and phenotypic transfer. Full article

The application of livestock manure can introduce antibiotic-resistant bacteria (ARB) into soil–vegetable systems, potentially shaping the antibiotic resistance profiles of plants. This study investigated the effects of manure containing chlortetracycline (CTC) on antibiotic resistance in pakchoi grown in three distinct soil types (black soil, fluvo-aquic soil, and red soil). The results demonstrated that CTC-manure application significantly increased the relative abundance of CTC-resistant endophytic bacteria (CREB), with the magnitude of the increase following the order: black soil (235.43%) > fluvo-aquic soil (64.5%) > red soil (10.68%). Furthermore, the presence of CTC promoted the proliferation of Acinetobacter and increased the abundance of potential pathogens (such as Klebsiella, Rhodococcus, and Corynebacterium), thereby elevating the risk of antibiotic resistance transmission. Metabolomic analysis revealed that CTC induced the most substantial metabolic alterations in pakchoi grown in red soil. Correlation analysis indicated that the CREB community structure was primarily shaped by soil properties, including total nitrogen, total phosphorus, and organic matter, and was significantly correlated with indigenous soil ARB (Pseudomonas, Bacillus, and Stenotrophomonas). This study elucidates the mechanisms underlying manure-driven antibiotic resistance dissemination in vegetable production systems and offers a theoretical foundation for developing agricultural practices to mitigate associated risks. Full article

Background/Objectives: Herein, we report the clinical cases of two affected first-degree relatives from a family with highly variable macular dystrophy, expanding the known phenotype spectrum with mutations in the thyroid hormone receptor beta gene (THRB). Methods: Multimodal retinal imaging included wide-field fundus photography, fundus autofluorescence (FAF), spectral domain optical coherence tomography (SD-OCT) imaging, performed alongside functional testing (visual fields, electroretinogram (ERG)), metabolic blood analyses, and genetic testing of both cases. Results: A 67-year-old female patient presenting with reading difficulties and visual impairment since childhood was referred for evaluation and counseling for potential treatment options. Extensive ophthalmologic examination, including multimodal retinal imaging and functional testing, revealed an occult macular dystrophy. Her 39-year-old son reported similar visual symptoms in combination with mild photophobia. In multimodal retinal imaging, he also showed a macular dystrophy but with a vitelliform phenotype. Genetic testing identified the heterozygous pathogenic variant c.283+1G>A in the thyroid hormone receptor beta gene (THRB) in both patients. Conclusions: This report shows a high intrafamilial variability of macular dystrophy caused by a heterozygous THRB mutation, which has only recently been recognized as a cause of macular dystrophy. Here, we describe a novel clinical presentation characterized by a vitelliform lesion, expanding the phenotypic spectrum of THRB-associated macular dystrophy. Full article

Stray cats (Felis vaga) are key hosts for feline and zoonotic pathogens. From June to August 2024, we conducted a cross-sectional study across six districts in Shenzhen, China, involving 126 cats sampled from three types of sites. Multiple specimens were tested via quantitative real-time PCR (qPCR) for feline coronavirus type I (FCoV-I), feline calicivirus (FCV), feline herpesvirus type I (FHV-I), feline panleukopenia virus (FPV), and rabies virus (RABV); serum was analyzed for RABV-neutralizing antibodies by the fluorescent antibody virus neutralization (FAVN) assay. The overall pathogen positivity was 89.68%. FPV was most prevalent (61.90%), followed by FCV (57.14%), FCoV-I (46.83%), and FHV-I (23.02%). No RABV nucleic acid was detected. The co-infection rate reached 62.70%, primarily dual infections (33.33%). Geographical variation was observed, with significantly higher FCoV-I in Longgang than Futian (p < 0.05). RABV seropositivity was only 6.00%. FCV and FPV co-occurred most frequently (Jaccard = 0.456). All pathogen pairs had relative risk (RR) > 1, suggesting non-random co-infections, though not significant after correction. In summary, major feline pathogens are widespread with frequent co-infections among Shenzhen stray cats, while low rabies immunity indicates potential public health risk. Targeted control measures are warranted. Full article

Fungi and bacteria associated with bark beetles can facilitate successful tree colonization, and, in some cases, these fungi act as pathogens of trees. The red turpentine beetle (RTB, Dendroctonus valens) is a bark beetle native to North America that colonizes stressed pines, rarely killing healthy trees. The fungal communities associated with RTB adults, larval galleries, and control tree phloem from red pine (Pinus resinosa) and white pine (P. strobus) forests in the Great Lakes region of the United States were characterized using both culture-independent and culture-dependent methods. Similarly, the bacterial communities associated with RTB adults in the same region were characterized using a culture-independent method. There were significant differences between the adult beetle fungal communities and the tree-based fungal communities. Culture-independent sequencing of RTB adults showed high abundances of the fungal order Filobasidiales (red pine: 28.71% relative abundance, white pine: 6.91% relative abundance), as well as the bacterial orders Enterobacterales (red pine: 53.72%, white pine: 22.15%) and Pseudomonadales (red pine: 15.86%, white pine: 12.91%). In contrast, we isolated high amounts of fungi in the orders Pleosporales (red pine: 21.79%, white pine: 15.90%) and Eurotiales (red pine: 15.38%, white pine: 16.51%) from the adult beetles by culturing. Culture-independent sequencing of beetle galleries yielded high abundances of fungi in the orders Helotiales (red pine: 22.23%, white pine: 23.21%), whereas culture-based isolation from the same galleries yielded high amounts of Eurotiales (red pine: 17.91%, white pine: 17.91%), Hypocreales (red pine: 16.42%, white pine: 16.42%), and Ophiostomatales (red pine: 23.39%, white pine: 23.39%). This contrasts with the culture-independent method, where, likely due to limitations in the sequencing method, the Ophiostomatales accounted for only around 2% of the fungi from RTB galleries in both pine species. We observed a high species-level diversity of Ophiostomatales associated with RTB, isolating 14 species from the Great Lakes region. Leptographium terebrantis, a species that has been described in association with RTB throughout the United States, was the most common species (e.g., >35% of the Ophiostomatales relative abundance in red pine environments and >14% of the Ophiostomatales relative abundance in the white pine environment). This study enhances our understanding of RTB-associated fungi and bacteria in the beetle’s native range at both the community and species levels. Full article

Introduction: Klebsiella variicola, a member of Klebsiella pneumoniae complex, has emerged as an opportunistic pathogen for human infection; however, antimicrobial resistance and hypervirulent characteristics of K. variicola have rarely been investigated in South Korea. Methods: We analyzed 76 clinical K. variicola isolates collected from 12 hospitals between September 2022 and October 2023. Bacterial identification was performed by MALDI-TOF MS. Antimicrobial susceptibility was tested by disk diffusion tests. Resistance determinants and virulence traits were investigated, and whole-genome sequencing was performed for hypermucoviscous or carbapenem-resistant K. variicola isolates. Results: Most (89.5%, 68/76) were susceptible to all 18 antimicrobials tested in this study, and 3 isolates harbored bla_CTX-M-15. One isolate carried bla_KPC-2 on its IncX3 plasmid, which is closely related to carbapenem-resistant K. pneumoniae plasmids. Capsular typing revealed 51 wzi allelic types. Ten isolates showed mucoid phenotype, mainly with KL60 and KL61. Conclusions: This study reveals relatively low resistance rates in K. variicola strains but the presence of multidrug-resistant and hypermucoviscous K. variicola strains. In addition, the evidence of interspecies dissemination of bla_KPC-2 highlights the need for continuous genomic surveillance. Full article

To address the limitation of traditional broad-spectrum antimicrobial agents in compromising skin microbiota homeostasis, this study developed Lactobacillus plantarum fermented milk (FM) as an innovative strategy for selectively regulating microbial communities to restore skin microbiota balance. FM was produced through protease hydrolysis in combination with L. plantarum fermentation. Selective antibacterial properties were evaluated via monoculture experiments (Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis) and pathogen–commensal co-culture systems. It was found that FM can selectively inhibit pathogens (E. coli and S. aureus) and promote the growth of commensal bacteria (S. epidermidis) in monoculture, and can reduce the growth and competitiveness of E. coli and S. aureus while relatively increasing the colony count of S. epidermidis in the co-culture system. Metabolomic profiling was further performed to identify metabolic alterations induced by FM. It was found that FM can activate the pyruvate metabolic node, significantly enhancing the metabolic fluxes of lactic acid, citric acid, and short-chain fatty acids, which triggered the acid stress response of pathogenic bacteria while consuming a considerable amount of energy, attenuating their reproductive capacity without impacting the growth of commensal bacteria. Overall, FM showed selective antimicrobial activity against pathogens (E. coli, and S. aureus) and preservation of commensal S. epidermidis, offering a foundational reference for the development of postbiotics aimed at maintaining cutaneous microbial homeostasis. Full article

Boxwood (Buxus sp.) plays a key role in historical gardens due to its evergreen foliage and resilience. However, recent outbreaks of disease caused by fungal pathogens such as Calonectria spp. (C. pseudonaviculata, C. henricotiae) and Pseudonectria spp. (P. buxi, P. foliicola), as well as pest pressures from Cydalima perspectalis, have led to significant losses. This study examined 100 boxwood plantings across the Czech Republic to evaluate pest and disease occurrence. Further, six modern boxwood cultivars from the groups of BetterBuxus^® and NewGen^® were tested in field trials under the climatic conditions of the Czech Republic, focusing on their resistance to abiotic stress and foliage color retention throughout the year. Laboratory trials confirmed all cultivars were susceptible to C. pseudonaviculata, with ‘Renaissance’ showing the slowest disease progression. Field assessments under two contrasting management regimes (“Minimalistic” and “Pampered”) indicated sporadic boxwood blight incidence but frequent Volutella blight outbreaks, particularly where plants suffered frost stress. Leaf color, an important esthetic trait, was evaluated using Munsell charts and measuring the relative chlorophyll content. ‘Skylight’ most closely matched Buxus sempervirens in the shade of green and winter color. Full article

Traditional diets based on diverse edible plants are increasingly threatened by climate change, which exposes crops to abiotic and biotic stressors such as drought, soil salinity, UV radiation, microorganisms, and insect herbivory. Understanding how these conditions influence both the nutritional and nutraceutical profiles, as well as the availability of key compounds, is essential to preserve their functional value. Piper auritum Kunth, used in Mexican gastronomy, was selected to assess two abiotic stress scenarios: drought stress (DS) and salicylic acid (SA) to simulate plant defense against pathogens and/or predators. We evaluated proximate composition, dietary fiber, total phenolics, total flavonoids, antioxidant capacity, low molecular weight carbohydrates (LMWCs), monomeric composition, and essential oil volatiles. Additionally, the simulated gastrointestinal digestion (INFOGEST) with an additional rat small-intestine extract (RSIE) revealed that both SA and DS shifted sugar distribution, especially for soluble and structural pools. SA treatment correlated with synthesis of secondary metabolites, particularly oxygenated and hydrocarbon terpenes. Both abiotic stressors modulated LMWC release during digestion, altering the distribution of sugars such as raffinose and galacturonic acid, with potential prebiotic implications. Essential oil analysis revealed stress-specific shifts in volatile composition, particularly in safrole, β-caryophyllene, and related terpenes. Beyond individual compound changes, the combined evaluation of composition, antioxidant properties, and volatile profile provides a comprehensive view of how abiotic stress can reshape the functional potential of P. auritum. To our knowledge, this is the first report on LMWC relative abundance across INFOGEST stages for a quelite species and on the integrated effect of DS and SA on its chemical profile. These findings highlight the importance of including compound release and functional traits, alongside chemical characterization, in future assessments of traditional plants under climate-related stress to safeguard their contribution to sustainable diets. Full article

Filoviruses are among the most lethal viral human pathogens known, with significant relevance to public health, yet their evolutionary history remains poorly resolved. This study applied a multiprotein molecular phylogenetic approach to investigate the evolutionary and temporal dynamics of the family Filoviridae. Amino acid sequences from the proteome and seven individual proteins (NP, VP35, VP40, GP, VP30, VP24, L) were analyzed using MEGA 12, with RelTime inference anchored on uniform calibrations, and integration of epidemiological data (cases, fatalities, case fatality). The phylogenetic reconstructions revealed robust topologies for most proteins, though selective pressures on GP, VP30 and VP40 generated more variable patterns. Temporal inferences supported the classification of filoviruses into three groups: an ancestral lineage (>1 MYA, fish- and reptile-associated), an intermediate lineage (BCE–1 MYA, bat-associated), and a contemporary lineage (CE, ebolaviruses and marburgviruses). VP30 and VP40 showed consistent associations with epidemiological outcomes in Orthoebolavirus zairense, suggesting their interplay may underlie enhanced dispersal and virulence. Contrariwise, Orthoebolavirus restonense emerged as a natural counterpoint for comparison with other potential human pathogenic filoviruses. Taken together, these findings highlight that filoviral evolution is intrinsically linked not only to viral biology but also to the ecology and history of their hosts. Full article