搜索结果：Journal of applied microbiology

The 2023 iteration of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) estimated prevalence, incidence, and health burden for 375 diseases and injuries, including 12 mental disorders. We assess past, current, and emerging trends in the prevalence and burden of mental disorders across sexes and age groups, for 21 regions, 204 countries and territories, and by Socio-demographic Index (SDI) quintile, from 1990 to 2023. Mental disorders included in GBD 2023 were anxiety disorders, major depressive disorder, dysthymia, bipolar disorder, schizophrenia, autism spectrum disorders, conduct disorder, attention-deficit hyperactivity disorder, anorexia nervosa, bulimia nervosa, idiopathic developmental intellectual disability, and a residual category of other mental disorders. A literature review identified epidemiological data for each disorder. These were analysed via a Bayesian meta-regression to estimate prevalence by disorder, sex, age, location, and year. Disorder-specific prevalence was multiplied by disability weights representing the severity of health loss associated with each disorder to estimate years lived with disability (YLDs). Deaths due to anorexia nervosa were assessed with a Cause of Death Ensemble modelling strategy to estimate deaths by sex, age, location, and year, and then multiplied by the standard life expectancy at age of death to estimate years of life lost (YLLs). YLDs equalled disability-adjusted life-years (DALYs) for all mental disorders except anorexia nervosa (the only mental disorder considered as an underlying cause of death in GBD), for which DALYs represented the sum of YLDs and YLLs. We presented prevalence, deaths, YLDs, YLLs, and DALYs as counts, age-specific rates per 100 000 population, and age-standardised rates per 100 000 population. We estimated 1·17 billion (95% uncertainty interval 1·06-1·31) prevalent cases of mental disorders globally in 2023, equivalent to an age-standardised prevalence rate of 14 210·7 cases (12 849·5-15 940·1) per 100 000 population. These estimates represented a 95·5% (75·0-121·2) increase in prevalent cases and 24·2% (11·4-41·4) increase in age-standardised prevalence rate between 1990 and 2023. All mental disorders showed increases in prevalent cases between 1990 and 2023, while notable increases were seen in age-standardised prevalence rates for anxiety disorders, major depressive disorder, dysthymia, anorexia nervosa, bulimia nervosa, schizophrenia, and conduct disorder. There were an estimated 171 million (127-228) DALYs due to mental disorders globally across sex and age in 2023, equivalent to an age-standardised DALY rate of 2070·5 DALYs (1519·1-2750·5) per 100 000 population. Mental disorders contributed to 6·1% (4·8-7·6) of all-cause DALYs in 2023, making them the fifth leading cause of global DALYs (up from 12th in 1990). DALYs were almost entirely composed of YLDs. Mental disorders were the leading cause of YLDs in 2023 (up from second in 1990), explaining 17·3% (14·8-20·6) of all-cause global YLDs. Leading causes of mental disorder DALYs were anxiety disorders (ranked 11th among the 304 diseases and injuries at Level 4 of the GBD cause hierarchy), major depressive disorder (15th), and schizophrenia (41st). Globally in 2023, mental disorder age-standardised DALY rates were higher among females (2239·6 [1643·7-3014·1] per 100 000) than among males (1900·2 [1399·8-2510·8] per 100 000), and peaked in the 15-19 years age group (2617·3 [1850·6-3696·8] per 100 000). All locations showed increased mental disorder DALY rates in 2023 compared with 1990, ranging across countries and territories from 1302·4 (952·7-1683·7) per 100 000 in Viet Nam to 3555·8 (2661·9-4715·0) per 100 000 in the Netherlands. Across SDI quintiles, DALY rates ranged from 1853·0 (1352·1-2469·3) per 100 000 for middle SDI to 2184·1 (1606·1-2890·3) per 100 000 for high SDI. A significant health burden was imposed by mental disorders in all countries and territories in 2023, irrespective of the health resources available. In some instances, this burden has increased over time and is unevenly distributed across populations. Stronger surveillance systems, particularly in low-income and middle-income countries, are required. Additionally, we need more coordinated and inclusive policies to reduce the burden through early treatment and prevention, tailored to sex and age differences across locations. Responding to the mental health needs of our global population, especially those most vulnerable, is an obligation, not a choice. Gates Foundation, Queensland Health, and University of Queensland.

The growing aging population and staff shortages are placing pressure on Dutch nursing homes (NHs). These challenges have led to an increased interest in digital health technologies. Among these are wearable devices that allow for remote continuous monitoring of vital signs. An example is the Healthdot (smartQare), a wearable electronic device that continuously monitors heart rate, respiratory rate, and physical activity. In the context of acute respiratory infections (ARIs) in NHs, where initial symptoms can go unnoticed, continuous monitoring may aid in early recognition, timely intervention, and reduce staff workloads. However, little is known about how health care professionals perceive the use of continuous vital signs monitoring devices, such as the Healthdot, for this cause in NHs. This study aims to explore the perspectives of healthcare professionals on the use of the Healthdot for early detection and monitoring of ARIs in NHs, to inform potential future implementation. Semistructured interviews were conducted with 20 physicians, nurses, and certified nursing assistants from 4 NHs and 1 acute geriatric community hospital located in a NH. Interview transcripts were thematically analyzed to identify themes regarding their perspectives on the use of the Healthdot for monitoring ARIs in this setting. Five main themes were identified that related to the appropriate use of the Healthdot for NH clients and health care professionals: alignment of Healthdot use and NH clients' treatment policies, balancing safety and freedom, impact of the Healthdot on work processes, supporting rather than replacing care, and possible use during pandemics and in the future. Additionally, several preconditions for the use of the Healthdot were identified, including its usability, a support base among care staff, adequate training and guidance, communication with NH clients and their relatives, and a clear policy regarding its use. Given the complexity of care in NHs, where clinical care is typically balanced against quality of life and a homelike environment, physicians generally expressed reserved attitudes toward the Healthdot, highlighting the need to consider multiple factors in its implementation. Care staff were generally positive about the device. Nevertheless, tailored assessment for each individual NH client remains essential, balancing treatment goals, safety, autonomy, and person-centered care. Additionally, clear communication and alignment between health care professionals in this setting are crucial, specifically regarding their expectations of the Healthdot's role in care processes. This study offers practical guidance that may inform future implementation efforts of continuous vital sign monitoring devices in NHs.

Post-mortem microbiology (PMM) is a significant diagnostic tool to identify infectious agents, unfortunately not always applied in forensic context. Two cases of meningo-encephalitis in juvenile females are reported. The first case involved an 8-year-old girl found dead in her bed, after a three-months clinical history of shortness of breath, coughing, vomiting, headache and purulent discharge from the ear canal. A diagnosis of adenoidal hypertrophy was made by pediatricians. At autopsy, acute and chronic multifocal inflammatory infiltrates were found at the brain, lungs, thymus and adenoidal tissue. The microbiological analysis performed on cerebrospinal fluid (CSF) was positive for Human Herpesvirus 6 (HHV-6). The cause of death was assessed as meningo-encephalitis based both on histological findings and PMM results. Based on the clinical history, medical malpractice issues were assessed. The second case is related to an 11-year-old girl's death, who showed a rapid neurological decline with cerebral edema and decompressive craniotomy, after a prompt diagnosis of otitis media and mastoiditis. Microbiological tests performed on samples taken before death and on formalin-fixed tissues taken after death, showed negative results. The cause of death was assessed as acute meningo-encephalitis based on the histological findings. They were mostly represented by wide inflammatory infiltrates within the cerebral parenchyma and subpial regions. Comparison of these two cases shows the strengths and limitations of PMM in the identification of the etiological agents. This issue can be crucial in the evaluation of infectious diseases and medical malpractice. In this regard, a reliable PMM requires that recommended sampling protocols be applied to biological fluids and soft tissues.

Candida albicans and Staphylococcus aureus polymicrobial biofilms cause chronic wound infections and tolerate standard antimicrobials poorly. Their endurance is driven by fungal hyphal morphogenesis, interkingdom co-aggregation, and virulence proteins, including SAP5, which create a strong polymicrobial biofilm. Homovanillic acid, a plant-derived phenolic molecule, was tested for its antibiofilm effects on hyphal growth, bacterial-fungal interactions, and SAP5-associated pathogenicity. Integrated in vitro, real-time dynamic, and in vivo methods were used to assess antibiofilm activity. Microtiter plate experiments evaluated the minimum biofilm inhibitory concentration (MBIC), whereas a real-time Biofilm Infection Simulator System examined biofilm formation under flow circumstances. SAP5 proteinase activity was measured spectrophotometrically, and microbiological and histological investigations confirmed in vivo efficacy in a catheter-associated rat wound model. HVA significantly inhibited polymicrobial biofilm initiation at sub-inhibitory concentrations, with an MBIC of 128 µg mL-1 corresponding to an 82.18% reduction in biofilm formation. Real-time analysis confirmed substantial suppression of biofilm development. Inhibition of C. albicans hyphal morphogenesis impaired S. aureus adhesion and destabilized biofilm architecture, while SAP5 activity was reduced by 53.37%. In vivo treatment promoted effective wound healing, marked by reduced inflammation, near-complete epithelial regeneration, and organized collagen deposition by day 14. HVA demonstrates potent multitarget antibiofilm activity and represents a promising plant-derived therapeutic candidate for managing polymicrobial biofilm-associated wound infections.

This study aimed to assess the effects that long-term exposure to sub-lethal concentrations of demethylation inhibitor (DMI) fungicides have on the population growth dynamics of the floral nectar yeast specialist Metschnikowia reukaufii. Synthetic populations of M. reukaufii were propagated for 112 days in presence of sublethal concentrations of imazalil (0.031, 0.125, and 0.5 μg mL-1) or no fungicide. ANOVA of aligned rank transformed colony forming unit (CFU) data showed that log10CFU values significantly depended on the treatment, the propagation step, and the interaction between these factors. However, hierarchical clustering of the series of log10CFU data did not reveal a clear differentiation of the control vs. imazalil treatments. Moreover, significant differences in median log10CFU were only observed in the first propagation step between the treatments containing 0.125 and 0.5 μg mL-1 of imazalil, and between some earlier vs. later propagation steps in the artificial nectars containing 0.031 and 0.5 μg mL-1 of imazalil (#2 vs. 24, and #1 vs. 4 and #1 vs. 24, respectively). Finally, continuous exposure to imazalil did not clearly select for increased resistance (i.e. higher minimum inhibitory concentration (MIC)) of M. reukaufii populations to DMI and non-DMI fungicides. The results of this study suggest that exposure to sublethal concentrations of imazalil might not have a significant long-term effect on the growth dynamics of M. reukaufii. However, given the ephemeral and highly dynamic nature of nectar microbial communities, even a short-term growth effect such as the one detected in the present study might be ecologically relevant.

Indwelling urethral catheters are the most widely used medical devices across the world, and catheter-associated urinary tract infections (CAUTIs) are the most common type of healthcare acquired infection. For many patients, urinary catheter blockage is a common and recurring problem, which can have considerable negative impact on patient health and well-being. Blockage primarily stems from the formation of crystalline bacterial biofilms on catheter surfaces, which can lead to upper urinary tract infection (UTI) and the onset of serious clinical complications. Potential solutions to this important clinical problem include the development of novel antibiofilm agents that can prevent formation of these communities on urinary catheters. However, traditional de novo methods of drug discovery are laborious, expensive, have long lead times and carry a high risk of failure in the clinical trial stages. One potential approach to mitigate this risk and cost, is the evaluation of pre-existing licensed drugs for those with useful antibiofilm or antimicrobial activity. Here we review current preclinical evidence for antibiofilm and antimicrobial activities in licensed drugs from a range of classes, such as urease inhibitors, Selective Serotonin Reuptake Inhibitors (SSRIs), phenothiazines, oncology therapeutics and non-steroidal anti-inflammatory drugs (NSAIDs). In doing so, we consider the application of the repurposing approach to control CAUTI and catheter blockage, and identify key challenges and opportunities related to delivery of repurposed drugs to the catheterised urinary tract.

People living with HIV (PLWH) may be exposed to harmful political, social, economic and environmental factors that exacerbate their risk of mental health conditions. Such factors can interact synergistically to worsen HIV and mental health-related outcomes, creating a syndemic. This study aims to review existing literature on mental health-related syndemics and their impact on HIV and mental health outcomes. CINAHL, Embase, MEDLINE, PsycInfo, Scopus and ProQuest were searched. We included observational studies that investigated a potential mental health-related syndemic and/or reported the impact of a syndemic on HIV outcomes (antiretroviral therapy [ART] adherence or viral suppression), or mental health outcomes (mental health-related quality of life, depression, anxiety, schizophrenia, bipolar disorder, post-traumatic stress disorder or psychological distress) among PLWH. Screening, data extraction and quality assessment were conducted by two independent reviewers. The Newcastle-Ottawa Scale (NOS) was used to assess the quality and risk of bias. The impact of syndemic count on ART adherence and viral suppression was pooled using random effects using STATA and the remaining findings were synthesised narratively. PRISMA guidelines were followed. 32 studies were included with sample sizes ranging from 51 participants to 14,261. Six studies reported on mental health-related syndemics among PLWH, four of which found depression or distress to be the most influential syndemic factor. Mental health conditions within syndemics often cluster and are significantly associated with socioeconomic factors such as food insecurity, stigma and violence. Fifteen of 16 studies found a significant association between adherence and the number of mental health-related syndemics. Pooled odds ratio of seven studies showed a significant reduction in adherence (OR = 0.73; 95% CI = 0.55 - 0.96); heterogeneity was high (I 2 = 98.58%). Eleven of 13 studies found a significant association between the number of mental health-related syndemics and being virally suppressed. Four studies resulted in a significant pooled odds ratio for having detectable viral load (OR = 1.26; 95% CI = 1.10 - 1.44); heterogeneity was moderate (I 2 = 52.38%). Despite wide variation in how syndemics were defined and measured across studies, our findings suggest that mental health conditions, particularly depression, strongly influence synergising syndemics among PLWH, and mental health-related syndemics negatively impact ART adherence and viral load. These findings underscore the need for syndemic-informed holistic care models to address the intersecting burden of mental health conditions and psychosocial factors among PLWH.

Recirculating aquaculture systems require high-throughput removal of pollutants during wastewater treatment, but their performance is often constrained by membrane fouling, which primarily results from the accumulation of bacterial biofilms. This study developed and evaluated a quorum quenching (QQ) strategy as an efficient and eco-friendly antifouling approach to prevent fouling of membrane modules in bioreactors. This study employed three-dimensional (3D) printing technology to immobilize a marine-derived QQ enzyme, YtnP, within an alginate matrix to fabricate QQ scaffolds designed to replace conventional feed-channel spacers in membrane modules. The physicochemical properties, durability, and antifouling performance of the resulting 3D-printed composite scaffolds were systematically evaluated. The optimal printing ink formulation comprised 0.025% purified YtnP and 11.11% sodium alginate at pH 7.2, followed by cross-linking in 2% CaCl₂ for 2 min. YtnP enzymes self-assembled and crystallized into nanoflower structures at the scaffold interface through calcium ion binding, demonstrating the role of calcium in facilitating both enzyme immobilization and alginate cross-linking. The QQ scaffolds maintained N-acyl homoserine lactone (AHL) removal efficiencies above 99.98% over nine consecutive runs and significantly suppressed biofouling on polyvinylidene fluoride (PVDF) membranes. The findings of this study enhance the feasibility of applying bioreactor technologies for aquaculture wastewater treatment and support the further development of recirculating aquaculture systems.

The long-term application of plant essential oil (EO)-based biocides is a popular "green" strategy for microbial control in heritage science. However, its ecological consequences in enclosed, humid environments remain poorly understood. This study investigated the unintended ecological shifts following prolonged use of an oregano-cinnamon EO mixture at the Wangjingmen earthen site, aiming to elucidate the mechanisms behind a subsequent macrofungal outbreak. We employed a multi-faceted approach combining environmental monitoring, high-throughput amplicon sequencing of bacterial (16S rRNA) and fungal (ITS) communities, and geochemical analysis. Our findings revealed that the stable, high-humidity microenvironment (>95% RH, 20-32°C), coupled with the selective pressure from continuous EO application, significantly altered the microbial community structure between 2022 and 2024. This led to a decrease in microbial richness and a community shift favoring resistant taxa. Specifically, bacterial communities shifted towards Pseudomonadota and Actinomycetota dominance, while fungal communities saw an enrichment of Ascomycota and Basidiomycota, culminating in the proliferation of Leucocoprinus at temperatures above 28°C. Geochemical analysis indicated that evaporation on vertical surfaces created nutrient and salt enrichment zones (e.g. NO₃⁻, SO₄²⁻, gypsum), which further facilitated fungal colonization. Predictive functional profiling suggested an enrichment of metabolic pathways related to EO degradation (e.g. limonene and pinene degradation), indicating microbial adaptation to the biocide. Prolonged application of EO-based biocides in stable, humid heritage environments can induce significant microbial dysbiosis, creating an ecological vacuum that facilitates opportunistic outbreaks of resistant macrofungi. Our findings highlight the potential risks of relying on broad-spectrum biocides without considering their long-term ecological impacts.

Glycerol dehydratase, encoded by dhaB, has been identified as the rate-limiting enzyme for biosynthesis of 1,3-propanediol (1,3-PDO). This study aimed to enhance the production of 1,3-PDO in Klebsiella pneumoniae by increasing the expression of dhaB. Plasmid-based and genomic integration systems for increasing the expression of dhaB were compared. The results showed that plasmid-promoted overexpression of dhaB resulted in growth inhibition, and no inhibition of cell growth was observed by overexpression of dhaB through genomic integration. However, overexpression by genomic integration, even though the copy number was higher, failed to increase 1,3-PDO production. Based on our data, the 1,3-PDO production was stimulated by moderate expression of dhaB through genomic integration. And then, increasing the supply of NADH improved the yield and production of 1,3-PDO from glycerol in K. pneumoniae to 0.71 mol/mol and 81 g/L, respectively, during a 2-L fed-batch fermentation process. Moderate overexpression of dhaB through genomic integration stimulated production of 1,3-PDO by K. pneumoniae.

Cholera outbreaks continue intermittently in Iraq. Recent outbreaks have highlighted the need for investigations. This study aimed to conduct an integrated epidemiological investigation of a cholera outbreak in Iraq using microbiological, molecular, immunological, and environmental approaches. This descriptive epidemiological study investigated the surveillance of acute watery diarrhoea (AWD) and environmental screening for Vibrio cholerae in 2024-2025. The microbiological and molecular characterisations, including virulence profiling, antimicrobial susceptibility, multilocus sequence typing (MLST), and herd immunity, were explored. A total of 136 cholera cases were identified in 2024, corresponding to an attack rate of 6.0 per 100 000 population. The outbreak was moderate in size, temporally restricted to August-November period, urban-centred, and affected both sexes of adults. All clinical isolates were toxigenic Vibrio cholerae O1 El Tor Ogawa and belonged to the distributed pandemic sequence type (ST) 69. Environmental detection was rare (2/103, 1.9%) and confined to non-toxigenic, genetically distinct isolates of sequence type (ST) 1994. Multidrug resistance was detected in 77.4% of clinical isolates, which predominantly exhibited moderate biofilm formation and were eliminated at low chlorine concentration (2 mg l⁻¹), whereas environmental isolates exhibited stronger biofilm formation and higher chlorine tolerance. Serological analysis indicated low population immunity (2.2%). No cholera cases were recorded in 2025. This cholera outbreak was moderate in size and showed predominantly human-mediated transmission in a low-immunity urban population. The clinical isolates were Vibrio cholerae ST69 and exhibited antibacterial resistance and biofilm production. These findings provided insights for early detection and control of future outbreaks.

Streptococcus suis is a serious zoonotic pathogen responsible for rapid progression and deadly infections in both humans and pigs. With an increasing number of reported cases and considering the limitations of standard routine identification, a simple, rapid, and cost-effective approach is needed. In this study, a label-free colorimetric assay based on gold nanoparticles (AuNPs) was applied with a specific aptamer, R8-su12. This assay offered simplified detection through observable color change, enabling visual analysis by the naked eye or assessment via UV-Vis spectrophotometry. Under the optimal assay conditions, the detection procedure was carried out within 45 min. The reaction of the aptasensor and other bacterial species, including Staphylococcus aureus, S. pneumoniae, S. pyogenes, Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecium, and E. faecalis, was not present, indicating the specificity of this assay. Moreover, the aptasensor exhibited high sensitivity with a limit of detection (LOD) at 1 CFU of S. suis and had broad reactivity with S. suis serotypes 1, 1/2, 9, and 14, as well as with S. suis isolated from clinical specimens. Thus, this aptasensor demonstrates proof-of-concept feasibility including clinical sample testing before practical implementation. It holds promise as a practical tool for the early screening and outbreak management of S. suis in a variety of settings, such as clinical laboratories, food safety, and the environment.

Diseases caused by pathogenic bacteria lead to severe economic losses every year in aquaculture. Aeromonas hydrophila (A. hydrophila) is responsible for a number of diseases in freshwater aquaculture and is regarded as a food-borne pathogen threatening human health. It is difficult to deal with bacterial infections due to the increase in antibiotic resistance in aquaculture. With the understanding of bacterial pathogenesis, developing drugs targeting bacterial virulence has been regarded as a useful approach to alternating antibiotics. In this study, the in vitro anti-virulence efficacies and in vivo therapeutic effect of apigenin were evaluated against A. hydrophila, and results found that apigenin could inhibit the activity of aerolysin (AerA) and production of quorum-sensing (QS)-regulated virulence factors at concentrations much lower than MIC. Molecular dynamics (MD) simulation results showed that apigenin could form a complex with AerA. Then we found that apigenin could block the oligomeric activity of AerA by forming an apigenin-AerA complex, and residues of Pro347 and Pro395 obtained by MD simulation were confirmed to be the main binding sites. The qPCR results demonstrated that apigenin could suppress the transcription of aerA and ahyR genes, which could lead to the decrease of QS-regulated virulence. Furthermore, apigenin could reduce AerA-induced cell injury and mitigate the mortality of channel catfish challenged with A. hydrophila. The findings partly clarified the mechanism of apigenin in dealing with A. hydrophila infections through anti-virulence strategy, and provided an approach for developing drugs in aquaculture.

Food contaminated with Salmonella remains a major cause of foodborne illness worldwide, posing a significant public health concern. Rapid detection of viable Salmonella in food is essential for effective food safety management. Viability PCR (vPCR) techniques using photo-reactive dyes such as PMA (Propidium monoazide) or PMAxx have emerged as promising approaches to suppress DNA amplification from membrane-compromised (dead) cells. However, most previous PMA-qPCR studies have mainly evaluated artificially contaminated meat samples, while their performance in naturally contaminated food matrices with complex microbiota remains limited. This study optimized a PMAxx real-time PCR assay for the detection of viable and viable but non-culturable (VBNC) Salmonella and evaluated its application in retail meat samples. The PMAxx real-time PCR technique effectively eliminated signals from 108 CFU/mL of dead cells in culture media samples. The limit of detection (LOD) was 10 CFU/g of viable cells in spiked chicken samples following 18 h of pre-enrichment in Buffered Peptone Water (BPW). Under 0.85% NaCl at 4 °C, Salmonella entered the VBNC state after 32 weeks, while PMAxx real-time PCR still detected signals. The method was evaluated using 33 chicken and pork samples collected from supermarkets and traditional markets. A total of 23 samples tested positive, including 13 samples detected after only 6 h of pre-enrichment. The results show 100% agreement with the traditional culture method performed in accordance with ISO 6579-1:2017. These findings indicate that the PMAxx real-time PCR assay can be applied for the rapid screening of viable Salmonella contamination in fresh food samples.

The number of published isothermal amplification assays has increased substantially in recent years. Unfortunately, no harmonized guidelines, such as the Minimum Information for Publication of Quantitative Real-Time PCR Experiments guidelines, exist for publishing these methods, often resulting in incomplete reporting of assay composition and performance. In this study, we systematically evaluated nine published loop-mediated isothermal amplification (LAMP) assays for the detection of Pseudomonas aeruginosa. We aimed to assess whether publications provide (i) sufficient information on assay composition to allow implementation and reproduction and (ii) robust data on assay performance to evaluate their applicability. Assays were screened for basic functionality, analytical specificity, sensitivity, and limit of detection (LOD) in head-to-head experiments with qPCR. Four assays lacked essential composition details, and almost all did not report DNA concentrations or replicate numbers. Only six assays consistently amplified target DNA. Analytical specificity testing with 19 non-target strains contradicted previously reported 100% specificity, with only 3 maintaining specificity above 90% in our evaluation. Sensitivity testing with 13 P. aeruginosa strains confirmed 100% sensitivity for two assays. However, LOD experiments revealed significantly higher values than originally reported, with qPCR outperforming all LAMP assays. These findings highlight substantial discrepancies between published data and real-world assay performance. The absence of standardized formats, consistent units, and complete methodological details undermines replicability. Although this study focused on P. aeruginosa, the identified issues are widely relevant across different microbial targets. We advocate for increased publication standards and quality controls to ensure transparency, utility, and comparability of isothermal amplification assays and to support their translation into clinical and environmental applications.IMPORTANCEOver the past decade, numerous isothermal amplification-based assays for the detection of pathogens or health-relevant microorganisms have been proposed, each claiming progress from the state-of-the-art and applicability to both clinical specimens and/or environmental samples. However, many published assays lack essential methodological details, and reported performance metrics are often inconsistent or incomparable. Using Pseudomonas aeruginosa as a representative model organism, we set out to test whether all details required for implementing assays were provided by original publications and if important assay characteristics were reproducible. The results of this systemic benchmarking of nine published loop-mediated isothermal amplification assays revealed major discrepancies between reported and experimentally measured performance in terms of specificity, sensitivity, and limit of detection. By highlighting the crucial elements that need to be reported, this work aims to improve the transparency, reproducibility, and overall quality of isothermal amplification assays, fostering their broader application across different research settings.

In India, no publicly funded seasonal influenza immunization is ongoing, partly because cost-effectiveness is yet to be established. We estimate the cost-effectiveness and budgetary impact of introducing seasonal influenza vaccination among adults aged ≥ 60 years in India, a known high-risk group. Estimates of disease burden including disability adjusted life years (DALYs) were generated from 3-year community cohorts and hospital-based study set up at four sites to estimate incidence of symptomatic acute respiratory infections (ARI), health care utilization, costs and outcomes and applied to India's 2021 estimated population. We used a decision analysis model from an abridged societal perspective for implementation of an inactivated trivalent influenza vaccination programme for older adults (aged ≥ 60 years) using either facility-based or outreach-based approaches. We estimated incremental cost-effectiveness ratio (ICER) per DALY averted and used India's per capita gross domestic product of US$ 2238 for 2021 as the cost-effectiveness threshold. We performed deterministic and probabilistic sensitivity analyses. We also estimated ICER for immunizing all those above 65 years or with comorbidities. Using financial cost of the vaccination programme and the direct medical cost of disease averted we estimated its budgetary impact. In 2021, older adults in India had 5.3 (95% CI: 3.6-7.7) million influenza-ARIs resulting in 36,149 (95% CI: 30,076-43,268) hospitalizations and 84,613 (95% CI: 39,895-155,454) deaths. It amounted to a total of 974,019 DALYs and US$ 66.6 million. As compared with no vaccination, facility-based and outreach-based influenza vaccination had an ICER of US$1979 and US$1851, respectively, below the cost-effectiveness threshold. In all sensitivity and scenario analyses, ICER estimates were below the threshold and vaccinating those above 60 years with specified comorbidities using a passive approach was found to be most cost-effective with least budgetary impact. Annually vaccinating older adults against influenza was found to be cost-effective in the majority of scenarios considered in the study. Given the budgetary implication, it might be prudent to focus on those with co-morbidities.

To investigate the in vitro antibacterial activity and related phenotypic effects of the novel antimicrobial peptide SAAP-148 against Helicobacter pylori (H. pylori). The minimum inhibitory concentration (MIC) of SAAP-148 against H. pylori strains, including multidrug-resistant isolates, was determined using the broth microdilution method. Its activity after short-term exposure to pH-adjusted conditions and its cytotoxicity toward gastric epithelial cells were evaluated. The effects of SAAP-148 on cell morphology, membrane permeability, established biofilms, bacterial viability, cell-associated urease activity, and ureA/ureB gene expression were assessed. Antibacterial activity against one four-drug-resistant H. pylori clinical isolate was further evaluated by colony counting and OD600 measurement. SAAP-148 exhibited potent antibacterial activity against H. pylori strains with different resistance profiles, with MIC values ranging from 16-32 µg/mL. SAAP-148 retained antibacterial activity after short-term exposure to pH-adjusted conditions and showed low cytotoxicity toward gastric epithelial cells. SAAP-148 treatment was associated with bacterial morphological damage, increased inner and outer membrane permeability, reduced biofilm biomass and biofilm-associated protein content, decreased cell-associated urease activity, and reduced ureA/ureB mRNA expression. In the dedicated MDR model, SAAP-148 also reduced viable counts of one four-drug-resistant H. pylori clinical isolate. SAAP-148 demonstrated promising in vitro activity against H. pylori and was associated with membrane disruption, antibiofilm effects, and reduced urease-associated readouts. These findings support further preclinical evaluation of SAAP-148 as a potential antimicrobial strategy against difficult-to-eradicate H. pylori.

This protocol details an optimized method for the production of small stable spheroids, their culture, and 3D imaging, for the study of the endothelial and insulin-producing β cells interactions in a 3D model of pancreatic islets. The 150-200 µm spheroids, mirroring the lowest range of islet sizes, were prepared from a selected ratio combining 1 intra-islet endothelial cells (MS-1 cells) to 20 insulin-secreting cells (β-TC-6). Staining, clearing, and mounting challenges of small spheroids and their tackling by employing low-melting point agarose and the CUBIC clearing technique are detailed, as well as key points for an efficient analysis of the 3D structure with different probes. Data indicate that NTPDASE-ectonucleotidase 3 does not colocalize with insulin in the spheroid model, suggesting varying maturity and functional levels of β-TC6 and that the complete procedure can also be applied to isolated pancreatic islets, with clear probing of intra-islet vessels. These findings underscore the effectiveness of the 3D imaging protocol in revealing complex pancreatic cell organization and interactions within the islet model.

The tuberculin skin test (TST), widely used for latent tuberculosis infection (LTBI) screening, shows reduced specificity in Bacillus Calmette-Guérin (BCG)-vaccinated individuals due to antigenic cross-reactivity. We aimed to quantify the impact of BCG vaccination on false-positive TST results and to identify key modifying factors. We systematically searched Embase, PubMed, Web of Science and the Cochrane Library up to 24 February 2025. A dual-comparison design was applied: head-to-head comparisons of interferon-γ release assay (IGRA) versus TST in BCG-vaccinated populations and between-group comparisons of TST positivity in vaccinated versus unvaccinated groups. Random-effects models were used to calculate risk differences (RDs) for false-positive results. A total of 242 studies were included, with 166 head-to-head (50 341 participants) and 176 between-group comparisons (456 429 participants). Both approaches yielded consistent estimates, including a head-to-head RD of 13% (95% CI 9-16%) and a between-group RD of 12% (95% CI 9-14%). False positivity decreased with increasing tuberculosis incidence and latitude. Immunosuppressed individuals showed lower RDs than general populations. BCG strain, age at vaccination and number of doses significantly influenced false positivity. We identified key modifiers of BCG-related TST false positivity. TST remains appropriate in high-incidence settings and for BCG specific strains, whereas IGRA may be preferred in low-incidence areas, in individuals vaccinated multiple times or in immunosuppressed individuals.