Arcobacter cryaerophilus is considered an emerging foodborne pathogen and is associated primarily with infectious gastrointestinal disease in humans. However, the underlying pathogenic mechanisms remain poorly understood. Therefore, the aim of the present study was to investigate the pathogenic potential of twelve A. cryaerophilus strains using various in vitro assays in two human colonic cell lines, HT-29/B6 and T84.
All strains tested were able to adhere to and invade into both cell lines, with strain-dependent differences in their adhesion and invasion rates. In addition, two strains showed cytotoxic effects on both cell lines. The ability to disrupt the epithelial barrier function of T84 cell monolayers was shown for two strains by measurement of transepithelial electrical resistance. As structural factors correlate with the barrier dysfunction, immunofluorescence staining of the tight junction domain was performed, and revealed an altered distribution of claudin-5 in infected cells.
The results highlight the strain-dependent pathogenic mechanisms of A. cryaerophilus that may contribute to key symptoms such as diarrhoea. These findings also highlight the importance of further research into the pathogen A. cryaerophilus.
© 2025. The Author(s).

This study explores the virulence mechanisms of Vibrio cholerae, with a particular emphasis on HapA, a zinc metalloprotease delivered via outer membrane vesicles (OMVs). The findings reveal that OMV-associated HapA disrupts the integrity of tight and adherens junctions in intestinal epithelial cell models more effectively than its purified counterpart, suggesting that association with OMVs substantially potentiates the pathogenic effects of HapA. The study further details the uptake of V. cholerae OMVs by epithelial cells, as well as their targeted degradation of key junctional proteins, including claudin, ZO-1, and β-catenin. These results highlight the critical role of OMV-associated HapA in compromising epithelial barrier function. Additionally, the use of spheroids and intestinal organoids in our experiments provides deeper insight into bacterial pathogenesis, offering valuable information for the development of targeted therapeutic strategies.
© 2025 The Author(s). Journal of Extracellular Vesicles published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles.

Epithelia are tightly connected cellular sheets, that shield our body from the external environment. They are continuously maintained by a pooled population of undifferentiated cells through differentiation. However, the maintenance mechanisms remain incompletely understood due to the difficulty of experimentally observing the differentiation process. To address this issue, we developed a culture method for long-term expansion of primary mammary basal cells with a set of compounds, that includes undifferentiated cells. An effective differentiation method regarding Claudin expression was also developed by simply removing compounds. To verify this differentiation-switching technique, we obtained microarray data comparing each differentiation state. Subsequent cellular analysis confirmed key transcription factors in each state: (1) EGR1 in undifferentiated basal cells is important for suppressing Claudin expression through maintaining the epithelial-mesenchymal transition (EMT) transcription factor TWIST1, (2) ELF3 in differentiated cells is important for actin organization and subsequent Claudin localization at the cell-cell border, that corresponds to the amount of GRHL3, an actin organizer. Their relevance was also observed in tissues and organoids. In summary, we present an effective tool for verifying molecular mechanisms that determine Claudin status in normal basal cell differentiation, that would be beneficial in epithelial cell biology, cancer biology, physiology, and regeneration research.
© 2025. The Author(s).

Giardia duodenalis is a widespread intestinal protozoan that affects mammals, including humans. Symptoms can range from being subclinical to causing severe abdominal pain and diarrhoea. Giardiasis often requires repeated treatment with synthetic drugs like metronidazole. In recent years, treatment failures in clinical cases involving nitroimidazoles have been increasingly reported. Consequently, identifying therapeutic alternatives is necessary. Medicinal plants have traditionally been used as antiparasitic compounds, but systematic evaluation under controlled experimental conditions is often lacking. Here, we evaluated the in vitro efficacy of Tabebuia avellanedae dry and hydroalcoholic extracts, as well as one of its active compounds, β-lapachone, as potential treatment against G. duodenalis infection. We observed effective antigiardial activity for all tested compounds, with β-lapachone exhibiting lower IC50 values than metronidazole. Cytotoxic effects often limit therapeutic concentration windows of opportunity, and choosing an informative model to assess them is not straightforward. In the present case, only T. avellanedae hydroalcoholic extract showed no cytotoxicity on tumoral human intestinal Caco-2 cell line, and only a trend of inhibition when tested on canine epithelial kidney MDCK cells. To introduce a more physiological test system, we used in vitro G. duodenalis infection experiments in a trans-well set-up using organoid derived monolayers (ODM) to assess at the same time drug efficacy against the parasite and safety on primary human intestinal epithelia, a likely surrogate for in vivo conditions. Our studies using this model point towards the potential therapeutic opportunity for non-systemic applications of T. avellanedae extracts and a relevant ingredient of these, β-lapachone. The data suggest that ODM co-cultures with G. duodenalis are suitable for testing antigiardial compounds, providing a more informative in vitro model before progressing to in vivo tests.
Copyright © 2025 The Authors. Published by Elsevier Ltd.. All rights reserved.

Metastasis is the leading cause of cancer-related deaths. Cancer-associated fibroblasts (CAFs) are abundant components within the tumour microenvironment, playing critical roles in metastasis. Although increasing evidence supports a role for small extracellular vesicles (sEVs) in this process, their precise contribution and molecular mechanisms remain unclear, compromising the development of antimetastatic therapies. Here, we establish that CAF-sEVs drive metastasis by mediating CAF-cancer cell interaction and hyperactivating TGF-β signalling in tumour cells. Metastasis is abolished by genetically targeting CAF-sEV secretion and consequent reduction of TGF-β signalling in cancer cells. Pharmacological treatment with dimethyl amiloride (DMA) decreases CAFs' sEV secretion, reduces TGF-β signalling levels in tumour cells and abrogates metastasis and tumour self-seeding. This work defines a new mechanism required by CAFs to drive cancer progression, supporting the therapeutic targeting of EV trafficking to disable the driving forces of metastasis.
© 2025 The Author(s). Journal of Extracellular Vesicles published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles.