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AnteayerPLOS ONE Medicine&Health

Exploratory study on the impact of <i>Ganoderma australe</i> extract on gut microbiota and immune gene expression in honey bees exposed to <i>Vairimorpha ceranae</i>

by Sarah Zuern, Bella Romero, Carlos Spichiger, Leandro Ortiz, Alejandro Jerez, Esteban Basoalto, Max Emil Schön, Sigisfredo Garnica

The microsporidium Vairimorpha (Nosema) ceranae is an emerging threat to honey bees (Apis mellifera), known to disrupt gut microbiota and suppress immune responses, potentially contributing to colony losses. Fungal extracts have recently gained interest as sources of bioactive compounds with antimicrobial and immunomodulatory potential. In this study, we explored the effects of different dietary supplements—sugar syrup, HiveAlive™, and a novel Ganoderma australe extract (GanoBee)—on gut bacterial composition and immune-related gene expression in honey bees subjected to experimental exposure to V. ceranae 1 x 104 spores per bee. The GanoBee diet altered the gut microbiota, notably reducing the relative abundance of Rhizobiaceae (Bartonella apis) and increasing Frischella compared to other treatments. While alpha diversity was not significantly affected by diet or exposure to V. ceranae, beta diversity differed significantly in bees fed with GanoBee. Additionally, the expression of the antimicrobial peptide genes abaecin and hymenoptaecin was elevated in both exposed and unexposed bees fed with GanoBee, depending on the sampling day. However, the establishment of V. ceranae infection appeared limited, likely due to low spore viability, and mortality in control bees was higher than expected. The low Vairimorpha ceranae infection levels observed in this study are likely attributable to reduced spore viability caused by storage conditions and/or suboptimal environmental conditions within the laboratory cages. Post hoc analyses indicated that the high viscosity of GanoBee-supplemented diets likely contributed to the elevated bee mortality observed, underscoring a critical limitation of the experimental design related to diet formulation and delivery method. These physical factors complicate the interpretation of treatment efficacy and highlight the importance of optimizing feeding protocols to avoid confounding effects. Despite these constraints, GanoBee demonstrated promising potential as a modulator of gut microbiota composition and immune-related gene expression, supporting the need for further research under improved and carefully controlled experimental conditions.

MetaMind: A multi-agent transformer-driven framework for automated network meta-analyses

by Achilleas Livieratos, Maria Kudela, Yuxi Zhao, All-shine Chen, Xin Luo, Junjing Lin, Di Zhang, Sai Dharmarajan, Sotirios Tsiodras, Vivek Rudrapatna, Margaret Gamalo

Background

Network meta-analysis (NMA) can compare several interventions at once by combining head-to-head and indirect trial evidence. However, identifying, extracting, and modelling these often takes months, delaying updates in many therapeutic areas.

Objective

To develop and validate MetaMind, an end-to-end, transformer-driven framework that automates NMA processes—including study retrieval, structured data extraction, and meta-analysis execution—while minimizing human input.

Methods

MetaMind integrates Promptriever, a fine-tuned retrieval model, to semantically retrieve high-impact clinical trials from PubMed; a multi-agent LLM architecture--Mixture of Agents (MoA)-- pipeline to extract PICO-structured (Population, Intervention, Comparison, Outcome) endpoints; and GPT-4o–generated Python and R scripts to perform Bayesian random-effects NMA and other NMA designs within a unified workflow. Validation was conducted by comparing MetaMind’s outputs against manually performed NMAs in ulcerative colitis (UC) and Crohn’s disease (CD).

Results

Promptriever outperformed baseline SentenceTransformer with higher similarity scores (0.7403 vs. 0.7049 for UC; 0.7142 vs. 0.7049 for CD) and narrower relevance ranges. Promptriever performance achieved 82.1% recall, 91.1% precision and an F1 score of 86.4% when compared to a previously published NMA. MetaMind achieved 100% accuracy on a limited set of remission endpoints regarding PICO (Population, Intervention, Comparator, Outcome) element extraction and produced comparative effect estimates and credible intervals closely matching manual analyses.

Conclusions

In our validation studies, MetaMind reduced the end-to-end NMA process to less than a week, compared with the several months typically needed for manual workflows, while preserving statistical rigor. This suggests its potential for future scaling of evidence synthesis to additional therapeutic areas.

Arp2/3 complex contributes to the actin-dependent uptake of <i>Aspergillus terreus</i> conidia by alveolar epithelial cells

by Natalia Mach, Julien Polleux, Lea Heinrich, Lukas Lechner, Iryna Levytska, Cornelia Lass-Flörl, Susanne Perkhofer

Aspergillus terreus is an opportunistic fungal pathogen associated with high mortality rates and intrinsic resistance to amphotericin B. Its ability to persist within host tissues without inducing strong immune responses was suggested to contribute to poor clinical outcomes. The cellular mechanisms underlying A. terreus interactions with host cells remain largely unexplored. In this study, we have used a micropattern-based infection model to investigate the early interactions between A. terreus conidia and alveolar epithelial cells, focusing on the role of Arp2/3-dependent actin remodeling. This system allows quantitative analysis of conidia-cell interactions under defined spatial conditions. We show that A. terreus conidia rapidly bind to micropatterned A549 cell islands, with conidial numbers increasing over time. Conidia were found in actin- and Lamp1-positive vesicles already after one hour of infection. Inhibition of the Arp2/3 complex significantly impaired conidial binding and disrupted the formation of actin-positive vesicles, confirming the essential role of Arp2/3-mediated actin remodeling in early stages of conidial uptake. A subset of conidia was localized to Lamp1-positive phagolysosomes and accumulated over time. Interestingly, we have identified a small but consistent population of Lamp1-positive vesicles decorated with actin structures, potentially resembling actin flashes. These structures were entirely abolished upon Arp2/3 inhibition, indicating active cytoskeletal remodeling at the phagolysosomal interface. Our findings provide the first mechanistic insights into A. terreus internalization by alveolar epithelial cells and establish Arp2/3-mediated actin dynamics as a key process in early host-pathogen interactions. This cellular pathway may further contribute to intracellular trafficking and help understand the delayed onset of A. terreus infections.

Human emotional odours influence horses’ behaviour and physiology

by Plotine Jardat, Alexandra Destrez, Fabrice Damon, Noa Tanguy-Guillo, Anne-Lyse Lainé, Céline Parias, Fabrice Reigner, Vitor H. B. Ferreira, Ludovic Calandreau, Léa Lansade

Olfaction is the most widespread sensory modality animals use to communicate, yet much remains to be discovered about its role. While most studies focused on intraspecific interactions and reproduction, new evidence suggests chemosignals may influence interspecific interactions and emotional communication. This study explores this possibility, investigating the potential role of olfactory signals in human-horse interactions. Cotton pads carrying human odours from fear and joy contexts, or unused pads (control odour) were applied to 43 horses’ nostrils during fear tests (suddenness and novelty tests) and human interaction tests (grooming and approach tests). Principal component analysis showed that overall, when exposed to fear-related human odours, horses exhibited significantly heightened fear responses and reduced interaction with humans compared to joy-related and control odours. More precisely, when exposed to fear-related odours, horses touched the human less in the human approach test (effect size: Rate Ratio(RR)=0.60 ± 0.24), gazed more at the novel object (RR = 1.32 ± 0.14), and were more startled (startle intensity – Cohen’s d = −0.88 ± 0.39; and maximum heart rate – Cohen’s d = 1.16 ± 0.47) by a sudden event. These results highlight the significance of chemosignals in interspecific interactions and provide insights into questions about the impact of domestication on emotional communication. Moreover, these findings have practical implications regarding the significance of handlers’ emotional states and its transmission through odours during human-horse interactions.

Phytochemical characterization, total phenolic and flavonoid content, antioxidant capacity, enzymatic profiling, and cytotoxicity of <i>Bidens pilosa</i> and <i>Croton</i> sp. from Colombia for applications in skin health

by Patricia Quintero-Rincón, Karina Caballero-Gallardo, Elkin Galeano, Oscar Flórez-Acosta

This study investigated the chemical and biological potential of Bidens pilosa and Croton sp., plants from megadiverse ecosystems in Colombia, collected in Santander de Quilichao (Cauca) and San Basilio de Palenque (Bolivar). The chemical profile was analyzed by UHPLC-ESI-Orbitrap-HRMS, and the total phenolic and flavonoid content was quantified using colorimetric methods. Antioxidant capacity was assessed using methods that evaluate reducing power and electron transfer mechanisms. The inhibition of key enzymes in skin aging, such as tyrosinase, hyaluronidase, and collagenase, was evaluated, as well as cytotoxicity in keratinocytes and human melanoma cells. Chemical characterization revealed distinctive phytochemical profiles: B. pilosa contained 21.1 mg GAE/g DT of phenolics and 64.6 mg RE/g DT of flavonoids, dominated by p-coumaric acid and rosmarinic acid, while Croton sp. exhibited higher levels of phenolics (169.4 mg GAE/g DT) and 54.1 mg RE/g DT of flavonoids, highlighting rosmarinic acid, p-coumaric acid and quercetin. Both extracts showed significant antioxidant capacity and enzyme modulation, including moderate collagenase inhibition (53.9–55.0%), high hyaluronidase inhibition (64.5–76.5%), and low tyrosinase inhibition (11.1–12.7%), suggesting protection of extracellular matrix and hyaluronic acid during skin aging. Sun protection factor was limited (SPF: 14.5 for B. pilosa and 11.6 for Croton sp.), with low ultraviolet absorption, consistent with low antityrosinase activity. Cytotoxicity assays demonstrated that B. pilosa was not toxic to HaCaT keratinocytes (IC₅₀ > 500 µg/mL) and displayed antimelanoma activity on A375 cells (IC₅₀ = 398.6 µg/mL), whereas Croton sp. showed moderate selectivity towards melanoma cells (IC₅₀ HaCaT = 329.5 µg/mL; IC₅₀ A375 = 189.0 µg/mL). The results suggest that both plants have potential in dermatological applications such as anti-melanoma agents, antioxidants, and modulators of skin aging enzymes, although highlight the importance of improving strategies to maximize their efficacy and safety.

Perspective of healthcare professionals on barriers and facilitators in exploring end-of-life care preferences of patients with pulmonary fibrosis: A qualitative study

by Lian Trapman, Lea M Dijksman, Jan C. Grutters, Saskia C.C.M. Teunissen, Everlien de Graaf

Background

Progressive pulmonary fibrosis is a lethal disease with a survival of 3–5 years with optimal medication treatment. Palliative care and advance care planning are therefore receiving increasing attention in the literature. However, structural implementation in clinical practice is still lacking. The aim of this study was to explore the needs, facilitators, and barriers for communication on the topic of end-of-life preferences of patients from the perspective of healthcare professionals.

Methods

A generic qualitative study was performed with focus groups and individual semi structured interviews with healthcare professionals. Data collection and analysis were performed iteratively. A thematic analysis was performed, following the methods of Braun and Clarke.

Results

Three focus groups and seven individual semi-structured interviews were conducted. Three themes were generated: (1) a lack of vision on palliative care, resulting in different approaches within the same clinic and showing the need for optimization of collaboration; (2) the importance of a learning-driven environment to support healthcare professionals skills and knowledge; and (3), the central role of the individual professional in developing skills and knowledge.

Conclusions/discussion

This study underscores the importance of behavioral and organizational change in palliative care to optimize conversations exploring values, preferences, and needs for end-of-life care for patients with pulmonary fibrosis. Leveraging the shared motivation of healthcare professionals to provide optimal care, integrating these findings into training and coaching programs can further enhance patient-centered approach in palliative care.

The EQo-Mental project: A protocol for a mixed-methods study on occupational balance and mental health in parents of children with developmental delays

by Desirée Valera-Gran, Miriam Hurtado-Pomares, Iris Juárez-Leal, Rocío Muñoz-Sánchez, Irene Campos-Sánchez, Paula Noce, Jessica Piñero, Eva-María Navarrete-Muñoz

Background

Parents of children with developmental delays (DD) often face significant challenges that affect their mental health and occupational balance. While early intervention services traditionally focus on child development, the occupational needs and well-being of parents remain underexplored. The EQo-Mental project aims to examine the association between parental mental health, occupational balance, and meaningful activity engagement, and to co-develop family-centred strategies that promote well-being in early intervention contexts.

Methods

This sequential mixed-methods study includes two phases. The quantitative phase will involve approximately 700 parents of children aged 0–6 years attending early intervention centres in Alicante, Spain. This phase comprises two components: (1) the psychometric validation of the Spanish versions of two occupational measures—the Occupational Balance Questionnaire (OBQ-E) and the Engagement in Meaningful Activities Survey (EMAS)—and (2) a cross-sectional analysis examining associations between occupational and mental health outcomes. Participants will complete a sociodemographic questionnaire along with validated self-administered instruments assessing occupational balance, meaningful activity engagement, stress, anxiety, depression, and psychological well-being. In the qualitative phase, participatory sessions and focus groups will be conducted with a subsample of parents and key stakeholders to explore perceived occupational and mental health needs and to co-design actionable strategies for improving occupational balance and family well-being. Participant recruitment began in November 2023 and is ongoing; data collection is expected to be completed by October 2025.

Analyses

Psychometric analyses will first be conducted to evaluate the validity and reliability of the OBQ-E and EMAS. Next, descriptive analyses and multiple regression models adjusted for potential confounders will be used to explore associations between occupational and mental health variables. Phase 2 consists of a participatory-action research process, including discussion groups and a multi-stakeholder focus group. Qualitative data will be analysed using reflexive thematic analysis.

Outcomes

Findings from EQo-Mental will inform the design of evidence-based, family-centred strategies that support occupational balance, parental well-being, and engagement in meaningful activities. By addressing the occupational needs of parents, the project seeks to foster more resilient families and strengthen early intervention services through an inclusive, occupation-focused approach.

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