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Neurovirol."],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Usutu virus (USUV) is a zoonotic arbovirus causing systemic and neuroinvasive diseases in birds and flu-like or sporadic neuroinvasive disease in humans. USUV-associated encephalitis is morphologically well-characterized in birds, while the mechanisms of cell-damage in spontaneously infected birds remain poorly understood. For the closely related West Nile virus, apoptosis significantly contributes to the pathogenesis of othoflavivirus-induced neuropathology. This study investigates the involvement of apoptosis in USUV-associated encephalitis in Eurasian blackbirds (\n                    <jats:italic>Turdus merula<\/jats:italic>\n                    ). To determine the apoptosis involvement, immunohistochemistry for cleaved-caspase-3 (CC3), TUNEL-assay, and RT-qPCR to compare mRNA expression levels of caspase 8 (extrinsic pathway) and caspase 9 (intrinsic pathway) were performed on brains of USUV-infected (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u200926) and USUV-uninfected (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u200916) blackbirds, as well as Africa 3 and Europe 3 USUV-lineages. The results showed a higher CC3 expression in USUV-infected (m\u2009=\u2009231,96\u2009\u00b1\u2009117,033) compared to uninfected blackbirds (m\u2009=\u200922,00\u2009\u00b1\u200913,31) (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.001). Similarly, higher TUNEL-positive cell counts were detected in USUV-infected (m\u2009=\u2009197,00\u2009\u00b1\u200983,56) compared to uninfected blackbirds (m\u2009=\u200919,00\u2009\u00b1\u200916,08) (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.01). No significant differences were detected comparing Africa 3 and Europe 3 lineages. Additionally, both caspases were upregulated in USUV-infected blackbirds. Caspase 8 was in average 4,37\u2009\u00b1\u20090,89 (CI \u03b1\u2009=\u20090,05) times higher in USUV-infected blackbirds (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.0001), while caspase 9 expression was in average 1,82\u2009\u00b1\u20090,89 (CI \u03b1\u2009=\u20090,05) times higher in USUV-infected blackbirds (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.01). Although both apoptotic pathways are involved, the extrinsic pathway is notably more active in USUV-infected blackbirds, highlighting its role in USUV-associated brain damage.\n                  <\/jats:p>","DOI":"10.1007\/s13365-025-01302-6","type":"journal-article","created":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T17:47:39Z","timestamp":1772473659000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Neuropathogenesis of Usutu virus-associated disease in Eurasian blackbirds (Turdus merula) involves apoptosis through the extrinsic pathway"],"prefix":"10.1007","volume":"32","author":[{"given":"Giuseppe","family":"Giglia","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gianfilippo","family":"Agliani","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bas B.","family":"Oude Munnink","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Reina S.","family":"Sikkema","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Teresa","family":"Mandara","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christine","family":"Fast","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marleen","family":"Gosens","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eefje J. 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