Agricultural systems require sustainable alternatives to chemical pesticides for controlling crop pathogens. This study evaluated biochar derived from grape pomace, a major winery byproduct, as an antimicrobial agent against tomato root pathogens (Fusarium oxysporum f. sp. lycopersici, Phytophthora infestans, Verticillium dahliae and Rhizoctonia solani). Under in vitro conditions, pathogens were exposed to biochar, washed biochar, and aqueous extracts in solid and liquid media. Biochar significantly inhibited fungal growth, whereas aqueous extracts showed no antifungal activity. Similar effects of washed and unwashed biochar indicate that inhibition is associated with the solid biochar structure rather than soluble compounds, and pH alone did not explain the effect. F. oxysporum and R. solani were most sensitive at lower concentrations. Metabolomic analysis identified 59 differential metabolites linked to stress responses. These findings highlight grape pomace-derived biochar as a promising sustainable strategy for managing tomato root pathogens.
Although acidification is widely used as a hurdle strategy to control foodborne pathogens, sublethal acid stress can enable pathogen survival and recovery, posing a food safety concern. This study assessed the impact of acid stress on foodborne pathogens and evaluated the efficacy of VERDAD N6 and ε-polylysine (ε-PL) against acid-stressed cells in broth and acidified cooked rice. Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, and Bacillus cereus (vegetative cells and spores) were screened for susceptibility to 0.5% acetic, lactic, and malic acids. Among the tested organisms, E. coli O157:H7 showed the highest tolerance to acetic acid, whereas B. cereus spores exhibited negligible reduction (<0.2 log). The combined antimicrobial effects of acetic acid, VERDAD N6, and ε-PL were further evaluated for E. coli O157:H7 and B. cereus, alongside ultrastructural analysis using transmission electron microscopy. In broth, 1% VERDAD N6 reduced growth rate, while 1% ε-PL resulted in complete inactivation. In acidified cooked rice (pH ≤ 4.6), acetic acid served as the primary hurdle against acid-stressed E. coli O157:H7. Supplementation with 0.5% VERDAD N6 and ε-PL at concentrations ≥0.01% further accelerated population decline during storage at 10 and 25 °C. In contrast, vegetative cells and spores of B. cereus showed minimal changes (<0.3 log). Microscopy revealed severe membrane disruption and ghost cell formation in E. coli O157:H7, whereas B. cereus exhibited limited structural damage. Overall, the combined treatment of acetic acid, VERDAD N6, and ε-PL was effective against acid-stressed E. coli O157:H7 but insufficient for controlling B. cereus spores.
Crop diversification through crop rotation or cover cropping is widely recognized as an important strategy to improve agroecosystem sustainability, enhance soil health, and suppress soilborne diseases. Rotating crops or introducing cover crops can disrupt pathogen life cycles, improve nutrient cycling, and promote beneficial microbes. However, the outcomes of diversification practices are often complex, influenced by soil type, crops, and pathogen pressures. Evaluating how cover crops and crop phase affect crop soilborne diseases and root-associated microbiome is critical for designing resilient cropping systems. This study evaluated the legacy effects of cover crops and crop phase on soybean root diseases and root-associated microbiome. Soybean plants were grown in soils collected from a corn-soybean rotation field experiment with and without cover crops, and then challenged with either Fusarium graminearum inoculum or soybean cyst nematode (SCN) in the growth chamber. Soils with a cover crop history significantly reduced F. graminearum-induced root rot, but had a limited impact on SCN, indicating divergent disease responses. Microbial profiling revealed that F. graminearum inoculum strongly reshaped bacterial communities, reducing Shannon diversity and enriching fast-growing copiotrophic taxa, including Bacteroidota genera (Pedobacter, Chitinophaga, Flavobacterium, and Mucilaginibacter) and Proteobacteria genera (Dyella, Pseudomonas, Rhizobium, and Paraburkholderia) regardless of cover crops. In contrast, SCN infection increased bacterial Shannon diversity in soybean-phase soils regardless of cover crops but decreased fungal Shannon diversity in soybean soils without cover crops, highlighting pathogen-specific microbial shifts. Cover cropping enhanced microbial heterogeneity under both pathogen pressures, enriching microbial taxa potentially involved in nutrient cycling (Chitinophaga and Mucilaginibacter), antagonism (Flavobacterium, Streptomyces, Pseudonocardia, and Nocardioides), and competitive interactions (Paraburkholderia). Correlation analyses further linked specific bacterial and fungal genera with disease suppression. Soilborne pathogens and cropping practices exerted interconnected, pathogen- and crop-specific effects on root microbial communities. Cover cropping offers a promising strategy to enhance microbial-mediated disease resilience in soybean systems, providing ecological insights into microbiome-driven plant health.
Bone defect repair presents a significant clinical challenge in oral and maxillofacial surgery and orthopedics. Implant-associated infection and insufficient osseointegration often compromise clinical outcomes, while traditional repair materials still struggle to simultaneously provide mechanical support, antibacterial activity, and osteogenic efficacy. This study employed magnetron sputtering to fabricate high-entropy nitride (HEN) coatings with varying silver (Ag) contents. Among these, the stable single-phase solid solution (TiZrNbHfTaAg)N coating with 5.7 at. percentage (at%) Ag exhibited outstanding comprehensive properties. This coating combines favorable biocompatibility with potent antibacterial activity, exhibiting antibacterial rates of 90.6% and 97.1% against common oral pathogens F. nucleatum and P. gingivalis, respectively. It also promotes osteoblast differentiation and mineralization by activating the MAPK/ERK signaling pathway, thereby upregulating the expression of genes and proteins associated with osteogenesis. In vivo validation using a rabbit tibia defect model showed that implantation of the (TiZrNbHfTaAg)N coating significantly improved indicators of bone regeneration and osseointegration strength. This coating offers a novel surface modification approach to address bone defect repair challenges through its "antibacterial-osteogenic" synergistic effect and bone regeneration mechanism mediated by the MAPK/ERK pathway, demonstrating considerable potential for clinical translation.
Transposable elements (TEs) are significant drivers of genome evolution, influencing the genome dynamics of clonal fungal pathogens such as those in the Fusarium oxysporum species complex (FOSC) that cause Fusarium wilt in over 100 plant hosts. Among these, Tropical Race 4 (TR4), a clonal lineage within the FOSC, poses a severe threat to global banana production. However, the contribution of TEs to genome variation and functional traits in TR4 remains poorly understood. Here, we investigated Helitron-associated structural variations in a TR4 strain from Mozambique (M1). This revealed two large deletions in core chromosomes associated with an active FoHeli1 Helitron transposon. One of these (464 kb) disrupted 151 genes, including the entire fusaric acid (FA) biosynthetic gene cluster, consequently abolishing FA production, altering secondary metabolite profiles, and increasing sensitivity to exogenous FA. Despite these metabolic changes, infection assays using wild-type, mutant, knock-out, and complemented strains demonstrated that FA production is dispensable for TR4 virulence in banana. Our study highlights the role of FoHeli1 in modulating the genetic and metabolic landscape of TR4, underscoring the broader impact of TEs on fungal genome evolution and functional diversification, especially in clonal lineages.
Early life exposure to common pathogens and a high pathogen burden during childhood can have long-term effects on immune development and overall health. These infections can trigger molecular changes, including alterations in gene expression and DNA methylation (DNAm), which regulate immune and metabolic pathways. Our aim was to identify biological processes underlying differential patterns of DNAm and gene expression in whole blood by infection status in European children. In the Rhea (Greece) and INMA (Spain) cohorts, serum/plasma samples collected at mean ages of 4 and 8 years were analyzed by multiplex serology to measure IgG against 14 antigens from 9 pathogens, and blood collected at a mean age of 8 years was used for DNAm and gene expression profiling. Epigenome- and transcriptome-wide analyses were conducted to assess association with childhood infections. A total of 290 unique CpGs were significantly associated with pathogen outcomes: 265 with seropositivity, 111 with first exposure timing, and one with viral burden. Cytomegalovirus (CMV) exposure accounted for the largest number of both epigenetic (n = 325) and transcriptomic (n = 8) associations. A total of 89 CMV-related CpGs had been described before in adults, and among novel ones, 54 showed consistent effects in adults. CMV-related CpGs were enriched for SUZ12 targets linked to morphogenesis, oxidative stress, and cognition. A previously developed CMV episcore in adults predicted serologically assessed CMV infection at 4 and 8 years of age, with area under the curve values ranging from 0.74 to 0.78 (95% CI 0.68-0.83). We identified novel DNAm and gene expression signatures of common childhood infections, particularly CMV, implicating immune and morphogenesis pathways. A subset of CMV-related DNAm signals showed consistent associations with those reported previously in adults, suggesting similar molecular effects across ages.
The oral-gut axis links the oral and gut microbiomes, both anatomically and functionally, and its dysregulation is implicated in periodontitis and inflammatory bowel disease (IBD). Bacterial extracellular vesicles (BEVs), nano-sized particles (20-250 nm) released by diverse microbes, serve as key mediators of inter-organismal communication along this axis. Carrying virulence factors, nucleic acids, and immunomodulators, BEVs influence microbial ecology and host immunity. In the oral cavity, pathogen-derived BEVs (e.g., from Porphyromonas gingivalis) promote biofilm formation, immune evasion, and tissue destruction. In the gut, BEVs from commensals (e.g., Akkermansia muciniphila) reinforce barrier function and suppress inflammation, whereas those from pathogens exacerbate dysbiosis. Critically, BEVs mediate bidirectional crosstalk: oral pathogen BEVs can translocate to the gut and worsen IBD, while beneficial gut-derived BEVs may attenuate periodontal inflammation. This review summarizes the roles of BEVs in oral-gut communication, their involvement in inflammatory disease pathogenesis, and their potential as biomarkers and therapeutics.
Influenza A, Influenza B, SARS-CoV-2, Respiratory Syncytial Virus and other respiratory pathogens are an ongoing public health concern. Rapid identification of these viruses early in infection is essential for effective treatment and outbreak control. The AMDI™ Fast PCR Mini Respiratory Panel (MRP) integrates sample preparation and real-time RT-PCR to detect Flu A, Flu B, SARS-CoV-2 and RSV from anterior nasal swab specimens in <10 min at the point of care. We established the analytical performance characteristics of the Fast PCR MRP and determined that the limit of detection (LoD) is 250 copies/mL for Flu A and RSV, and 500 copies/mL for Flu B and SARS-CoV-2. A reproducibility study at 3 clinical sites demonstrated ≥ 98.2% positivity for targets in a weak positive sample, ≥99.6% positivity for targets in a moderate positive sample and ≥99.3% negativity for a negative sample. Fast PCR MRP had 100% analytical reactivity for all strains tested (23 Flu A, 6 Flu B, 8 SARS-CoV-2 and 6 RSV) ≥ 98% predicted in silico inclusivity and no cross-reactivity to 40 microorganisms, nor interference from 15 endogenous and exogenous substances. The Fast PCR MRP delivers excellent analytical performance comparable to high complexity laboratory assays, at the point of care.
Recent advances in xenotransplantation have gained substantial public and clinical attention as genetically modified porcine organs are now being transplanted into living human recipients. While only case reports have been published to date, the first clinical trials for kidney xenotransplantation are now ongoing. This transition to clinical practice presents multiple implementation challenges for establishing scalable transplant programs while ensuring patient safety. Machine perfusion is expected to play a critical role in addressing these challenges by serving as a central platform for organ preservation, assessment, transport, and therapeutic intervention. Given the limited number of designated pathogen-free (DPF) breeding facilities, regional and international organ transport depends on robust preservation strategies during transit. Additionally, perfusion devices enable essential pre-transplant screening for zoonotic pathogens, a crucial safety measure unique to xenotransplantation. Further, given recent developments that allow for multi-day perfusion of grafts, wild-type grafts could potentially be genetically modified while being perfused ex situ. Beyond these perfusion modalities of isolated whole organs, machine perfusion offers a new therapeutic approach for patients with acute liver failure. Here, cross-circulation between a perfused genetically modified porcine organ and the patient can provide temporary liver replacement therapy. This mini-review summarizes the transformative potential of machine perfusion technology in clinical xenotransplantation with a focus on livers.
Antimicrobial resistance (AMR) is a major global public health threat, undermining the efficacy of commonly used antibiotics. Resistance patterns differ across bacterial taxa, including Enterobacteriaceae, non-fermenting Gram-negative bacilli, and Gram-positive cocci. This study aimed to provide a comparative analysis of antimicrobial susceptibility among reference strains with defined susceptible and resistant phenotypes, alongside selected clinical isolates, to evaluate the preservation of phenotypic traits and the impact of antibiotic use. Reference susceptibility strains exhibited high susceptibility across most antibiotics, whereas resistant reference strains demonstrated multidrug resistance. Among Enterobacteriaceae, reference strains harboring ESBL and AmpC mechanisms displayed resistance to penicillins, cephalosporins, and carbapenems. Non-fermenters, including Pseudomonas aeruginosa and Acinetobacter baumannii, showed both intrinsic and acquired resistance to multiple classes, particularly carbapenems and fluoroquinolones. Gram-positive cocci largely retained susceptibility to glycopeptides and linezolid, while MRSA, high-level aminoglycoside-resistant enterococci, and penicillin-resistant Streptococcus pneumoniae posed significant therapeutic challenges. Comparative analysis revealed that antimicrobial susceptibility is influenced not only by bacterial taxonomy but also by patterns of uncontrolled or inappropriate antibiotic use. Clinical strains of Klebsiella pneumoniae and Streptococcus pneumoniae displayed reduced and more variable susceptibility compared to the predictable profiles of reference strains. These findings highlight the importance of continuous surveillance, strict adherence to antimicrobial stewardship, and the use of standardized reference strains to ensure reliable susceptibility testing. Early detection of emerging resistance patterns is essential to guide effective therapy and mitigate the public health impact of multidrug-resistant pathogens.
Reptiles often inhabit environments that are in close proximity to humans and livestock, creating opportunities for parasite transmission. They are common in areas where they find shelter, food and warmth. The Bengal monitor lizard (Varanus bengalensis), a member of the family Varanidae, represents one of the largest groups of extant poikilothermic predators. Monitor lizards are known to harbor several tick species that serve as vectors for a variety of pathogens. No prior information is available in the literature regarding ticks infesting V. bengalensis in Pakistan as well as regarding the occurrence of Toxoplasma gondii in these ticks. Therefore, we aimed to determine the molecular prevalence of T. gondii in Amblyomma gervaisi ticks (n = 93) collected from 24 V. bengalensis in Buner District, Khyber Pakhtunkhwa Province, Pakistan, between May and September 2023. Polymerase chain reaction (PCR) amplified a 300 bp fragment specific for the ITS-1 region of T. gondii in 10 of the 93 (11%) A. gervaisi ticks. DNA sequencing and BLAST analysis confirmed the presence of T. gondii. Phylogenetic analysis showed that these sequences clustered with the ITS-1 sequences of T. gondii detected in reptiles and mammals from Pakistan, Brazil, China, Tunisia and Portugal. The prevalence of T. gondii in A. gervaisi was not limited to a specific tick sex, feeding stage or month of sampling. However, among the tick developmental stages, nymphs had the highest rate of T. gondii infection. In conclusion, for the very first time from Pakistan, we are reporting the presence of T. gondii in A. gervaisi that were infesting monitor lizards. We recommend that similar and large scale studies should be conducted in all those areas of Pakistan that are unexplored for the presence of T. gondii in A. gervaisi ticks. Prevalence of this parasite should also be screened in all the animals harboring these as well as other tick species. This will help in better understanding of T. gondii transmission to new hosts that will lead toward its effective control.
Allexiviruses (family Alphaflexiviridae) are widespread pathogens of vegetatively propagated allium crops, but their occurrence has not previously been documented in Ukraine. We surveyed cultivated allium plants collected in eight Ukrainian regions (2022-2025) and screened their samples for garlic virus B (GarV-B), garlic virus C (GarV-C) and shallot virus X (ShVX) using enzyme-linked immunosorbent assay (ELISA). GarV-B, GarV-C and ShVX were detected in 39/108 (36.1%), 23/108 (21.3%) and 21/108 (19.4%) plants, respectively, with infections which were strongly host-associated: garlic (n = 63) had high frequencies of indicated viruses (GarV-B-61.9%; GarV-C-36.5%; ShVX-28.6%), whereas onion samples (n = 33) were largely negative (ShVX-3.0%; GarV-B and GarV-C-not detected). Co-occurrence analysis within garlic revealed a nested allexivirus module in which GarV-C and ShVX occurred only in GarV-B-positive plants. RT-PCR and Sanger sequencing generated 11 partial genomes representing GarV-B, GarV-C, ShVX, GarV-A and GarV-D. Maximum-likelihood phylogenies placed Ukrainian allexivirus isolates within established global diversity and indicated both European- and Asian-affiliated lineages. These findings provide the first evidence of allexiviruses in Ukrainian allium crops, and support their inclusion in plant health surveillance and planting-material certification.
Dysgonomonas spp. is a microorganism that is occasionally isolated in patients with gastroenteritis, although its clinical relevance remains uncertain. It has also been reported in association with other underlying conditions, especially in immunocompromised individuals. However, the exact role of Dysgonomonas spp. requires further investigation. Our objective was to link the pathogenesis of Dysgonomonas spp. with the clinical context in pediatric patients, evaluating its significance in gastroenteritis and related diseases. This was a retrospective and descriptive prevalence study involving the isolation of Dysgonomonas spp. from the fecal samples of pediatric patients presenting with digestive symptoms (from September 2019 to December 2023). The strains were isolated on a selective medium for Campylobacter spp., identified using the MALDI-TOF method, and their antibiotic sensitivity was assessed. Cases in which other gastrointestinal pathogens were isolated alongside Dysgonomonas spp. were excluded. Fifty cases of patients with Dysgonomonas spp. infection were analyzed, of which 45 (41 D. capnocitophagoides; four D. gadei) met the inclusion criteria. Among these, 77.8% were isolated episodes of gastroenteritis, whereas 22.2% were cases of persistent gastroenteritis (PG). Eighty percent (80%) of the cases occurred in the context of associated pathologies. All strains were sensitive to clindamycin and co-trimoxazole. In cases of alteration of the intestinal microbiota, usually related to specific underlying pathologies and other concomitant factors, Dysgonomonas spp. may act as an opportunistic pathogen.
The PE_PGRS gene family in Mycobacterium tuberculosis exhibits extensive sequence variability across genotypes, which is consistent with antigenic divergence. Here we investigate how Mtb-despite lacking horizontal gene transfer-balances genomic stability with adaptive plasticity. Comparative analysis of 88 bacterial genomes reveals that PE_PGRS genes exhibit features facilitating mutability, including a significantly elevated CGGC tetramer density (mean 4.97 per 100 nt; range 1.7-7.4) compared with the genome-wide average (1.62 per 100 nt; p = 0.011) and depleted in out-of-frame stop codons, potentially conferring robustness to 1-nt and 2-nt frameshifts. Computational predictions suggest that CGGC motifs may promote secondary DNA structures, potentially destabilizing replication and contributing to replication errors, while the scarcity of out-of-frame stop codons allows continued translation beyond frameshifts, leading to changes in protein sequence and length. This dual organization may contribute to the observed adaptability of M. tuberculosis and could highlight a broader principle by which some pathogens evolve under strong constraints on horizontal gene transfer. We propose that CGGC-rich regions may function as programmed mutational hotspots across a wide range of microorganisms.
We report a case of fungal endophthalmitis caused by the rare fungus Fusicolla species in a 32-year-old female presenting with redness and pain in the right eye. The case illustrates the challenges in diagnosing infectious uveitis and highlights the importance of molecular diagnostics in identifying rare fungal pathogens. This is the first documented case of ocular involvement associated with this species.
To investigate the impact of mouth breathing (MB) on the salivary microbiome in children by comparing the differences in biochemical indicators, immune proteins and microbial community structure between mouth-breathing children and nose-breathing children, as well as among mouth-breathing children with different malocclusion types. Saliva samples were collected from 30 mouth-breathing children (MB group) and 10 nose-breathing children (control group), aged 8 to 12, between August 2023 and August 2024. The MB group was further subdivided into Angle Class I, II and III malocclusion subgroups, with 10 cases each. Ion concentrations were measured using an automatic biochemical analyser, pH was determined using a pH metre, immune protein levels were assayed by enzyme-linked immunosorbent assay and the structure and diversity of the salivary microbiota were analysed using 16S rRNA high-throughput sequencing. Compared with the control group, the MB group showed no significant difference in salivary pH (P > .05), but a significantly lower chloride ion (Cl⁻) concentration (P < .05). Conversely, the concentrations of secretory immunoglobulin A (SIgA) and peroxiredoxin-5 (PRDX5) were significantly higher in the MB group (P < .05). No statistically significant differences in these indicators were observed among the different malocclusion subgroups. Spearman correlation analysis revealed a positive correlation between PRDX5 and SIgA concentrations (rs = 0.808, P < .001) and negative correlations between both PRDX5 and SIgA concentrations and Cl⁻ concentration (rs = -0.588 and -.600, respectively; P < .001) in the MB group. Microbiota analysis indicated that the species richness (Chao1 index) of the salivary microbiota was significantly higher in the MB group. At the genus level, the relative abundances of Dialister, Streptobacillus, Anaeroglobus and Scardovia were significantly increased in the MB group (P < .05). MB alters the physicochemical properties of children's saliva, triggering an enhanced local immune response and a state of oxidative stress. This leads to dysbiosis of the salivary microbial community, characterised by a higher abundance of pathogenic bacteria associated with dental caries and periodontal disease and shows a correlation with pathogens linked to upper respiratory tract inflammation. These findings suggest that MB may impact oral and potentially systemic health by disrupting the oral microenvironment.
Serious fungal diseases are significant yet long neglected public health problem in India, setting an estimated 4.1% of the population and with the mortality from invasive fungal diseases approaching 50%, surpassing several nationally prioritised conditions. The emergence and spread of drug resistant pathogens such as Candida auris, azole resistant Aspergillus fumigatus, and Trichophyton indotineae together with COVID-19 associated mucormycosis epidemic, expose critical gaps in preparedness prevention and care despite the presence of a WHO collaborating centre and an Indian Council of Medical Research (ICMR) mycology Advanced Mycology Diagnostic and Research Centre network, major deficiencies persist in surveillance, diagnostics, antifungal access, stewardship, workforce, capacity and coordinated research OBJECTIVES: The Indian Fungal Infection National Declaration (I- FIND) seeks to catalyse a structured, time bound national response that elevates fungal diseases to a recognised public health priority and measurably reduces morbidity, mortality, and fungal resistance. Its objectives are to identify and target key at risk populations (including those with immunosuppression, critical illnesses, chronic lung disease and poorly controlled diabetes, to acknowledge major community mycoses such as chronic dermatophytosis, vulvovaginal candidiasis, and fungal keratitis, and to define clear domains for action across governance, surveillance, diagnostics, clinical management, research, workforce development, public awareness, One Health, and accountability. I- FIND identifies 9 core domains, including governance, surveillance, diagnostics, clinical management, research, workforce development, public awareness, One Health measures, and accountability. Proposals cover creating a National Task Force, issuing a five-year fungal disease control strategy, integrating mycology network, setting diagnostic standards, incorporating stewardship and prioritizing translational research funding. The declaration is supported by major Indian and international professional societies guides formal adoption and implementation across India.
As the global population ages rapidly, delaying and preventing age-related diseases have become urgent priorities in public health and biomedical research. During aging, mitochondrial dysfunction is a core molecular hallmark and a common pathogenic mechanism underlying multiple age-related disorders. Age-related mitochondrial dysfunction typically manifests as diminished metabolic capacity, impaired organelle renewal, and disrupted redox homeostasis. These factors interact to form a feedback loop constraining mitochondrial adaptability. Specifically, the interdependent decline in NAD+ availability, impaired mitochondrial biogenesis, and excessive oxidative stress render single-pathway interventions ineffective in mitigating systemic functional impairments triggered by aging. To address this complex mechanism, this review presents a novel tri-axis anti-aging model encompassing three key compounds: nicotinamide mononucleotide/nicotinamide riboside (NMN/NR), pyrroloquinoline quinone (PQQ), and l-ergothioneine (EGT). Within this framework, NMN/NR serves as a broad NAD+-dependent regulator of mitochondrial homeostasis, with its most immediate effects on metabolic activation, while PQQ and EGT may further strengthen mitochondrial remodeling and redox resilience, respectively. While each compound has distinct functional emphases, they are highly mechanistically coupled, collectively forming a closed-loop network regulating mitochondrial number, function, and homeostasis. This review synthesizes preclinical and emerging clinical evidence supporting the standalone or combined use of NMN/NR, PQQ, and EGT across various diseases. Collectively, by conceptualizing mitochondrial aging as a systemic imbalance rather than isolated molecular defects, this paper highlights a three-axis model of NMN/NR, PQQ, and EGT. This framework offers a theoretical foundation for mitochondrial-targeted anti-aging interventions while laying the groundwork for future clinical research, nutritional interventions, and the development of multi-target combination strategies.
The mechanisms leading to the formation of sclerotic lesions in focal segmental glomerulosclerosis (FSGS) remain incompletely understood; however, podocyte detachment and loss are considered key pathogenic events. Ubiquitin-specific protease 40 (USP40) is a deubiquitylating enzyme expressed in podocytes. In the present study, we investigated the role of USP40 in podocytes, focusing on its impact on the adhesion molecule integrin β1, which is essential for anchoring podocytes to the glomerular basement membrane. When USP40 knockout mice were subjected to an experimental FSGS model, they exhibited significantly more severe proteinuria and glomerulosclerosis than control mice, along with a marked reduction in podocyte number and integrin β1 expression. Consistently, knockdown of USP40 in cultured podocytes resulted in decreased integrin β1 expression and impaired adhesive properties compared with sham-treated cells. In HEK293 cells transfected with ubiquitin constructs, USP40 suppressed integrin β1 monoubiquitylation. In a separate internalization assay, USP40 prevented the clathrin-mediated endocytosis of integrin β1. In USP40 knockout mice, clathrin-coated vesicles colocalizing with integrin β1 were more frequently observed in podocyte foot processes than in control mice. Together, these findings suggest that USP40 functions as a deubiquitylating enzyme that stabilizes integrin β1 at the podocyte plasma membrane by preventing its endocytosis. We therefore propose that the USP40-integrin β1 axis represents a potential therapeutic target for FSGS.
Cutibacterium acnes (C. acnes) is the most common pathogen in shoulder periprosthetic joint infection (PJI) but remains difficult to distinguish from contamination due to its low virulence and presence as a skin commensal. Diagnostic ambiguity and variable antibiotic resistance further complicate management. We analyzed 87 C. acnes isolates from patients undergoing evaluation for shoulder PJI across two institutions. Isolates were obtained from intraoperative cultures and stored for subsequent analysis. Hemolysis, aerotolerance, and growth rate were assessed in vitro. Antimicrobial susceptibility to five antibiotics was determined using minimum inhibitory concentration (MIC) testing, with interpretive breakpoints defined by both the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. Infection status was classified per the 2018 International Consensus Meeting (ICM) criteria into definite, probable, possible, or unlikely categories. Associations between phenotypic traits and infection classification were assessed using univariate and multivariable statistical models. Only 12.6% (n = 11) of cases met criteria for definite infection, while the majority (48.3%) were classified as probable. Hemolysis and aerotolerance were observed in 51.7% and 54.0% of isolates, respectively, and were strongly associated with one another (P < .001); however, neither phenotype was associated with infection classification in univariate or multivariable analysis. Clindamycin resistance was observed in a minority of isolates but varied by standard, with 12.6% classified as resistant by EUCAST and 6.9% as non-susceptible by CLSI. No isolates were resistant to vancomycin; doxycycline and rifampin MICs were uniformly low. Neither hemolysis nor aerotolerance reliably distinguished definite PJI from non-definite cases, limiting their diagnostic utility in this cohort. Clindamycin resistance was uncommon, yet classification differed based on the interpretive criteria used (CLSI vs. EUCAST), highlighting potential inconsistencies in clinical reporting. Moreover, the predominance of "probable" infections illustrates a critical limitation of current consensus criteria, which may both underestimate true infection burden in indolent presentations and overestimate infection due to reliance on positive cultures in the absence of clear clinical findings. More precise diagnostic frameworks may help address current classification challenges and better support clinical decision-making in shoulder arthroplasty.