安特卫普热带医学研究所

This study aims to characterize the phenotypic behavior and in vivo persistence of a ceftriaxone-tolerant Neisseria gonorrhoeae clinical isolate from a single patient and evaluate the potential role of tolerance in treatment failure. A previously identified ceftriaxone-tolerant vaginal isolate was compared with isogenic and clinical non-tolerant strains. Bacterial growth was assessed in vitro, and tolerance was quantified using the minimum duration required to kill 99% of the population (MDK99), and persistence was evaluated in an in vivo Galleria mellonella infection model. Whole-genome sequencing (WGS) and transcriptomic (RNA-sequencing [RNA-seq]) profiling were performed to identify tolerance-associated genetic and transcriptional signatures. The tolerant strain exhibited prolonged MDK99 values across ceftriaxone concentrations, persisting for up to 24 hours under drug exposure. It also showed delayed early-phase growth, suggesting a fitness cost. In vivo, the tolerant strain remained viable up to 8 hours after treatment, whereas non-tolerant strains were cleared. WGS revealed identical gene content across all isolates, but non-synonymous mutations in pilE_3, a type IV pilin gene, were exclusively present in tolerant strains. RNA-seq analysis showed upregulation of pilin-associated genes and downregulation of zinc-independent ribosomal paralogs (rpmE2 and ykgO), suggesting a combined mechanism of surface remodeling and translational suppression associated with the tolerant phenotype. Ceftriaxone tolerance enables prolonged survival of N. gonorrhoeae despite apparent susceptibility by standard MIC-based testing. This phenotype may contribute to treatment failure, recurrent infection, and ongoing transmission, indicating the need for revised diagnostic and therapeutic strategies.IMPORTANCECeftriaxone remains the last reliable option for gonorrhea therapy, yet recurrent infections can occur despite isolates being classified as susceptible by MIC testing. One possible explanation is antibiotic tolerance, a phenotype that allows survival during drug exposure without changes in MIC. Although tolerance has been described in other pathogens, its role in gonococcal infection has remained poorly defined. In this study, we provide the first detailed characterization of a ceftriaxone-tolerant Neisseria gonorrhoeae clinical isolate associated with repeated treatment failure. By combining in vitro killing assays, an in vivo Galleria mellonella infection model, whole-genome sequencing, and transcriptomic profiling, we demonstrate that tolerance enables prolonged survival under ceftriaxone and is linked to pilin gene variation and ribosomal remodeling. These findings illustrate how a clinically observed phenomenon can be mechanistically dissected and emphasize tolerance as a hidden factor contributing to gonococcal persistence and potential treatment failure.

Background:Migrants and ethnic minorities (MEM) often face health challenges and structural and social barriers in accessing primary care. Pharmacists, as accessible healthcare professionals with expertise in medication management can address some of these barriers, however, their roles in serving MEM remain underexplored. Objectives:This study explored the adaptive communication and care practices of community pharmacists in responding to MEM clients in Flanders, Belgium. Methods:We conducted 42 h of non-participant observation and short reflective consultations with pharmacy staff of seven community pharmacies in two cities during February and March 2025. Field notes were structured using a semi-structured topic guide covering duration and content of pharmacist-client interactions; modes of communication; and delivery of pharmaceutical care. Framework analysis was used to code and categorize field notes according to micro-, meso-, and macro-level factors identified as influencing interactions between pharmacy staff and MEM clients. Results:Community pharmacy staff bridge healthcare gaps for MEM clients through multiple strategies, shaped by the dual logics of retail and healthcare. They were observed to accommodate diverse languages and proficiency levels; support medication and health (systems) literacy; address socio-economic challenges; and show sensitivity towards socio-cultural dynamics of care-seeking. Reflecting pharmacists' personal and professional motivations, these adaptive practices are shaped by meso-level factors such as staff composition, pharmacy layout, and social norms, as well as pharmacy-level discretionary actions within the Belgian health system and migration policy contexts. Conclusions:While well positioned to guide MEM clients through a fragmented health system, community pharmacists require systemic policy support to serve diverse population needs effectively.

Background Historically, the Democratic Republic of the Congo (DRC) has faced the greatest public health burden from mpox, including more than 70 000 probable cases from 1 January 2024 to 2 February 2025. However, there has been a relative paucity of investigation focused on mpox community engagement in DRC, including assessments of disease knowledge and risk perception.Methods Given the ongoing Clade I mpox public health emergency of international concern, and the linkage between sustained human-to-human transmission and dense sexual networks, we sought to investigate mpox knowledge and sexual behaviours among key populations. Between 20 March 2024 and 25 August 2024, we recruited 2794 participants distributed across Kinshasa, Kwango and North Kivu provinces, with a focus in urban centres where mpox risk was considered high.Results Most participants were considered other at-risk populations (948; 33.9%), followed by men who have sex with men (MSM, 828; 29.6%) and sex workers (897; 32.1%). Mpox knowledge, including transmission routes as well as sexual and health-seeking behaviours, was evaluated through questionnaires led by peer educators. Overall, only 6.1% of all participants reported prior mpox knowledge. Among this participant subset, zoonosis (‘direct contact with infected animals’) and ‘people living in high-risk areas’ were the most frequently selected options in regard to mpox transmission and populations at risk, respectively. When considering at-risk behaviours for mpox, those who identified as sex workers reported significantly higher risk sexual activities, including multiple sexual partners (80.3% of sex work participants), engaging in transactional sex (84.7%) and anonymous sex (80.8%) compared with MSM. However, both sex workers (44.8%) and MSM (56.7%) reported the highest health seeking behaviours for a suspected sexually transmitted infection.Conclusions Our results highlight the need to evaluate knowledge in high-risk communities, especially in an endemic setting. Community engagement, which incorporates both mpox knowledge and risk perception activities, is inclusive of at-risk populations and is needed for ongoing mpox containment and mitigation efforts.

Objective This study aimed to determine the prevalence and factors associated with pre-diabetes and undiagnosed type 2 diabetes (UDD) in Cambodia.Design This analysis used data from the WHO World Health Survey Plus, which was collected using a cross-sectional design with a GIS-based, three-stage sampling approach. Multiple logistic regression was used to identify key associated factors, based on a significance level of p<0.05.Setting Data were collected from all 25 provinces in Cambodia between 12 March 2023 and 31 May 2023.Participants 4427 individuals aged 18 years or older, residing in the selected household for at least 6 months in the past year.Primary outcome measures Pre-diabetes (Haemoglobin A1c (HbA1c) 5.7%–6.4%) and UDD (HbA1c≥6.5%), without prior knowledge of having type 2 diabetes (T2D).Results The weighted prevalences of pre-diabetes and UDD were 26.4% (95% CI 24.0% to 29.0%) and 9.3% (95% CI 7.9% to 11.0%). Pre-diabetes prevalence was higher in urban areas compared with rural areas (adjusted OR, aOR=1.2, 95% CI 1.0 to 1.4), males (aOR=1.7, 95% CI 1.3 to 2.3), individuals aged 40–49 (aOR=1.8, 95% CI 1.4 to 2.4), individuals aged 50+ years group (aOR=2.9, 95% CI 2.3 to 3.6) compared with the 18–39 years group, overweight individuals (aOR=1.7, 95% CI 1.4 to 2.0), obese (aOR=2.1, 95% CI 1.5 to 3.0) and those with elevated total triglycerides (aOR=1.3, 95% CI 1.1 to 1.5). Similar risk factors were identified for UDD, with the addition of hypertension (aOR=1.6, 95% CI 1.3 to 2.0) and high waist circumference (aOR=2.0, 95% CI 1.5 to 2.7).Conclusions The high prevalence of pre-diabetes and UDD in Cambodia is a pressing public health concern. Urgent and intensive interventions are needed to effectively prevent and manage T2D, particularly among urban residents, older persons and individuals with metabolic risk factors.

Rooted in long-standing assumptions and adapted from classifications mainly used for Latin American cutaneous leishmaniasis (CL), the nationally recommended clinical categories in Ethiopia for CL remain limited to localized cutaneous leishmaniasis (LCL), mucocutaneous leishmaniasis (MCL), and diffuse cutaneous leishmaniasis (DCL). However, these categories are associated with immune mechanisms which have not been validated in the Ethiopian context and thus risk misrepresenting the true clinical and immunopathological diversity. In this opinion piece, we will therefore outline key knowledge gaps and challenges in current clinico-immunological research on Ethiopian CL. In Ethiopia, Leishmania aethiopica is often assumed as the causative agent of these ‘LCL’, ‘MCL’ and ‘DCL’ forms, yet significant gaps in knowledge urge caution. For example, adoption of this ‘LCL’, ‘MCL’ and ‘DCL’ terminology and the associated immune mechanisms has led to inconsistent results. Most immunological studies on Ethiopian CL have focused on peripheral blood, resulting in little information about immune processes in the lesion, the original site of infection. Adding to the complexity, other species (including L. major, L. donovani and L. tropica as well as L. aethiopica hybrids) also circulate, with reports of Leishmania RNA virus co-infection. To address these challenges, we propose a multidimensional approach that combines standardized clinical documentation with appropriate lesion and blood sampling for in-depth profiling of both immune responses and parasite diversity. Adopting this holistic approach will require inclusive (inter)national collaborations with a focus on promoting equitable biobank and data sharing which will strengthen local research capacities. By addressing the main challenges and knowledge gaps in current clinico-immunological research through a multidimensional approach, this opinion aims to provide the tools to achieve a better and unbiased understanding of the immunopathogenesis of Ethiopian CL, a severe yet under-investigated disease. Such progress is essential for improving CL management in Ethiopia and aligns with the World Health Organization priority on controlling CL.