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Treating this fungal infection conventionally has significant limitations, prompting the search for alternative therapies. In this context, fungal extracellular vesicles (EVs) hold relevant potential as both therapeutic agents and targets for the treatment of fungal infections. To explore this further, we conducted a study using pharmacological inhibitors of chitinase (methylxanthines) to investigate their potential to reduce EV release and its subsequent impact on fungal virulence in an in vivo invertebrate model. Our findings revealed that a subinhibitory concentration of the methylxanthine, caffeine, effectively reduces EV release, leading to a modulation of H. capsulatum virulence. To the best of our knowledge, this is the first reported instance of a pharmacological inhibitor that reduces fungal EV release without any observed fungicidal effects.<\/jats:p>","DOI":"10.3390\/jof9111052","type":"journal-article","created":{"date-parts":[[2023,10,27]],"date-time":"2023-10-27T03:33:51Z","timestamp":1698377631000},"page":"1052","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Traversing the Cell Wall: The Chitinolytic Activity of Histoplasma capsulatum Extracellular Vesicles Facilitates Their Release"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9285-3670","authenticated-orcid":false,"given":"Alessandro F.","family":"Valdez","sequence":"first","affiliation":[{"name":"Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"},{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461, USA"},{"name":"Division of Infectious Diseases, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taiane Nascimento","family":"de Souza","sequence":"additional","affiliation":[{"name":"Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"},{"name":"Division of Infectious Diseases, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jhon Jhamilton Artunduaga","family":"Bonilla","sequence":"additional","affiliation":[{"name":"Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Zamith-Miranda","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461, USA"},{"name":"Division of Infectious Diseases, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alicia Corbellini","family":"Piffer","sequence":"additional","affiliation":[{"name":"Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"},{"name":"Unit\u00e9 Biologie des ARN des Pathog\u00e8nes Fongiques, D\u00e9partament de Mycologie, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, F-75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5478-9006","authenticated-orcid":false,"given":"Glauber R. S.","family":"Araujo","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biof\u00edsica de Fungos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2856-9667","authenticated-orcid":false,"given":"Allan J.","family":"Guimar\u00e3es","sequence":"additional","affiliation":[{"name":"Instituto Biom\u00e9dico, Departamento de Microbiologia e Parasitologia\u2014MIP, Universidade Federal Fluminense, Niter\u00f3i 24210-130, RJ, Brazil"},{"name":"Rede Micologia, RJ, FAPERJ, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5875-9886","authenticated-orcid":false,"given":"Susana","family":"Frases","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biof\u00edsica de Fungos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"},{"name":"Rede Micologia, RJ, FAPERJ, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7190-9471","authenticated-orcid":false,"given":"Alana Kelyene","family":"Pereira","sequence":"additional","affiliation":[{"name":"Instituto de Qu\u00edmica, Universidade Estadual de Campinas, Campinas, S\u00e3o Paulo 13083-970, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taicia Pacheco","family":"Fill","sequence":"additional","affiliation":[{"name":"Instituto de Qu\u00edmica, Universidade Estadual de Campinas, Campinas, S\u00e3o Paulo 13083-970, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3946-8375","authenticated-orcid":false,"given":"Igor L.","family":"Estevao","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, University of Texas El Paso, El Paso, TX 79902, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-6935-8847","authenticated-orcid":false,"given":"Angel","family":"Torres","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, University of Texas El Paso, El Paso, TX 79902, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Igor C.","family":"Almeida","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, University of Texas El Paso, El Paso, TX 79902, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9905-5754","authenticated-orcid":false,"given":"Joshua D.","family":"Nosanchuk","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461, USA"},{"name":"Division of Infectious Diseases, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9281-6856","authenticated-orcid":false,"given":"Leonardo","family":"Nimrichter","sequence":"additional","affiliation":[{"name":"Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"},{"name":"Rede Micologia, RJ, FAPERJ, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1900","DOI":"10.1080\/21505594.2022.2137987","article-title":"Pathogenicity & virulence of Histoplasma capsulatum\u2014A multifaceted organism adapted to intracellular environments","volume":"13","author":"Valdez","year":"2022","journal-title":"Virulence"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.idc.2015.10.009","article-title":"Histoplasmosis","volume":"30","author":"Wheat","year":"2016","journal-title":"Infect. 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