{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T01:34:14Z","timestamp":1792114454448,"version":"4.4.0"},"reference-count":29,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,6,6]],"date-time":"2019-06-06T00:00:00Z","timestamp":1559779200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,6,6]],"date-time":"2019-06-06T00:00:00Z","timestamp":1559779200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["097437\/Z\/11\/Z"],"award-info":[{"award-number":["097437\/Z\/11\/Z"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>HRT3 <jats:italic>in vivo<\/jats:italic> confocal microscopy (IVCM) images may indicate clinical outcome, but few studies have analysed this in fungal keratitis (FK). Adults with FK (diameter \u22653\u2009mm) presenting to Aravind Eye Hospital, India from 2012-3 were enrolled prospectively. IVCM was performed at baseline, days 7, 14 and 21 post-enrolment (+\/\u2212 3 days where possible). Specific morphologies were identified in IVCM images by a grader masked to microbiology and clinical outcome (defined as good: healed\/improving, or poor: enlarged ulcer, perforation or transplant\/glue). Associations with final visit outcome assessed using logistic regression. 143 FK participants were enrolled; 87 had good outcome, 56 had poor outcome. Poor outcomes were associated with stellate interconnected cellular processes with no visible nuclei (OR 2.28, 95% CI: 1.03\u20135.06, p\u2009=\u20090.043) in baseline IVCM images, and fungal filaments (OR 6.48, 95% CI:2.50\u201316.78, p\u2009&lt;\u20090.001) or honeycomb distribution of inflammatory cells (OR 5.24, 95% CI: 1.44\u201319.06, p\u2009=\u20090.012) in final visit images. Fungal filaments (OR 3.61, 95% CI:1.64\u20137.95, p\u2009=\u20090.001), stromal dendritiform cells (OR 2.88, 95% CI:1.17\u20137.11, p\u2009=\u20090.022), or stellate cellular processes with no visible nuclei (OR 2.09, 95% CI:1.14\u20133.82, p\u2009=\u20090.017) were associated with poor outcome if not in baseline but present in final visit images. IVCM can reveal morphological changes associated with clinical outcome.<\/jats:p>","DOI":"10.1038\/s41598-019-44833-9","type":"journal-article","created":{"date-parts":[[2019,6,6]],"date-time":"2019-06-06T10:03:09Z","timestamp":1559815389000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Cellular morphological changes detected by laser scanning in vivo confocal microscopy associated with clinical outcome in fungal keratitis"],"prefix":"10.1038","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5438-2858","authenticated-orcid":false,"given":"Jaya D.","family":"Chidambaram","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Namperumalsamy V.","family":"Prajna","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Srikanthi","family":"Palepu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shruti","family":"Lanjewar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manisha","family":"Shah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shanmugam","family":"Elakkiya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prajna","family":"Lalitha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Macleod","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew J.","family":"Burton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,6,6]]},"reference":[{"key":"44833_CR1","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1016\/S1542-0124(12)70291-4","volume":"7","author":"A Labbe","year":"2009","unstructured":"Labbe, A. et al. Contribution of in vivo confocal microscopy to the diagnosis and management of infectious keratitis. Ocul Surf 7, 41\u201352 (2009).","journal-title":"Ocul Surf"},{"issue":"11","key":"44833_CR2","doi-asserted-by":"publisher","first-page":"2285","DOI":"10.1016\/j.ophtha.2016.07.009","volume":"123","author":"Jaya D. Chidambaram","year":"2016","unstructured":"Chidambaram, J. D. et al. Prospective Study of the Diagnostic Accuracy of the In Vivo Laser Scanning Confocal Microscope for Severe Microbial Keratitis. Ophthalmology, https:\/\/doi.org\/10.1016\/j.ophtha.2016.07.009 (2016).","journal-title":"Ophthalmology"},{"issue":"Suppl 10","key":"44833_CR3","doi-asserted-by":"publisher","first-page":"S61","DOI":"10.1097\/ICO.0000000000000540","volume":"34","author":"V Kocaba","year":"2015","unstructured":"Kocaba, V., Colica, C., Rabilloud, M. & Burillon, C. Predicting Corneal Graft Rejection by Confocal Microscopy. Cornea 34(Suppl 10), S61\u201364, https:\/\/doi.org\/10.1097\/ICO.0000000000000540 (2015).","journal-title":"Cornea"},{"key":"44833_CR4","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1097\/00003226-199701000-00006","volume":"16","author":"K Winchester","year":"1997","unstructured":"Winchester, K., Mathers, W. D. & Sutphin, J. E. Diagnosis of Aspergillus keratitis in vivo with confocal microscopy. Cornea 16, 27\u201331 (1997).","journal-title":"Cornea"},{"key":"44833_CR5","doi-asserted-by":"publisher","first-page":"581","DOI":"10.1007\/s00417-007-0719-x","volume":"246","author":"W Shi","year":"2008","unstructured":"Shi, W., Li, S., Liu, M., Jin, H. & Xie, L. Antifungal chemotherapy for fungal keratitis guided by in vivo confocal microscopy. Graefe\u2019s Archive for Clinical and Experimental Ophthalmology 246, 581\u2013586, https:\/\/doi.org\/10.1007\/s00417-007-0719-x (2008).","journal-title":"Graefe\u2019s Archive for Clinical and Experimental Ophthalmology"},{"key":"44833_CR6","doi-asserted-by":"publisher","first-page":"1129","DOI":"10.3109\/02713683.2014.902073","volume":"39","author":"M Hovakimyan","year":"2014","unstructured":"Hovakimyan, M. et al. Morphological analysis of quiescent and activated keratocytes: a review of ex vivo and in vivo findings. Curr. Eye Res. 39, 1129\u20131144, https:\/\/doi.org\/10.3109\/02713683.2014.902073 (2014).","journal-title":"Curr. Eye Res."},{"key":"44833_CR7","doi-asserted-by":"publisher","first-page":"602","DOI":"10.1016\/j.ajo.2010.06.045","volume":"150","author":"EY Tu","year":"2010","unstructured":"Tu, E. Y. & Joslin, C. E. Recent outbreaks of atypical contact lens-related keratitis: what have we learned? Am. J. Ophthalmol. 150, 602\u2013608 e602, https:\/\/doi.org\/10.1016\/j.ajo.2010.06.045 (2010).","journal-title":"Am. J. Ophthalmol."},{"key":"44833_CR8","unstructured":"Srinivasan, M. et al. Guidelines for the Management of Corneal Ulcers at Primary, Secondary & Tertiary Care health facilities in the South-East Asia Region. 36 (2004)."},{"key":"44833_CR9","doi-asserted-by":"publisher","first-page":"982","DOI":"10.1136\/bjo.2009.175083","volume":"94","author":"SC Hau","year":"2010","unstructured":"Hau, S. C. et al. Diagnostic accuracy of microbial keratitis with in vivo scanning laser confocal microscopy. British Journal of Ophthalmology 94, 982\u2013987, https:\/\/doi.org\/10.1136\/bjo.2009.175083 (2010).","journal-title":"British Journal of Ophthalmology"},{"key":"44833_CR10","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1016\/j.ajo.2018.03.010","volume":"190","author":"Jaya D. Chidambaram","year":"2018","unstructured":"Chidambaram, J. D. et al. In vivo confocal microscopy cellular features of host and organism in bacterial, fungal and Acanthamoeba keratitis. Am. J. Ophthalmol., https:\/\/doi.org\/10.1016\/j.ajo.2018.03.010 (2018).","journal-title":"American Journal of Ophthalmology"},{"key":"44833_CR11","doi-asserted-by":"publisher","first-page":"638","DOI":"10.3928\/1081597X-20091111-01","volume":"26","author":"JO Croxatto","year":"2010","unstructured":"Croxatto, J. O., Tytiun, A. E. & Argento, C. J. Sequential in vivo confocal microscopy study of corneal wound healing after cross-linking in patients with keratoconus. J. Refract. Surg. 26, 638\u2013645, https:\/\/doi.org\/10.3928\/1081597X-20091111-01 (2010).","journal-title":"J. Refract. Surg."},{"key":"44833_CR12","doi-asserted-by":"crossref","unstructured":"Masters, B. R. & Kino, G. S. In Noninvasive diagnostic techniques in ophthalmology. (ed Masters, B. R.) Ch. 10, 152\u2013171 (Springer-Verlag, New York, 1990).","DOI":"10.1007\/978-1-4613-8896-8_10"},{"key":"44833_CR13","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2415-4-16","volume":"4","author":"GK Vemuganti","year":"2004","unstructured":"Vemuganti, G. K., Reddy, K., Iftekhar, G., Garg, P. & Sharma, S. Keratocyte loss in corneal infection through apoptosis: a histologic study of 59 cases. BMC Ophthalmol. 4, 16, https:\/\/doi.org\/10.1186\/1471-2415-4-16 (2004).","journal-title":"BMC Ophthalmol."},{"key":"44833_CR14","first-page":"291","volume":"48","author":"GK Vemuganti","year":"2000","unstructured":"Vemuganti, G. K., Sharma, S., Athmanathan, S. & Garg, P. Keratocyte loss in Acanthamoeba keratitis: phagocytosis, necrosis or apoptosis? Indian J. Ophthalmol. 48, 291\u2013294 (2000).","journal-title":"Indian J. Ophthalmol."},{"key":"44833_CR15","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1016\/j.exer.2014.01.004","volume":"120","author":"SD Hanlon","year":"2014","unstructured":"Hanlon, S. D., Smith, C. W., Sauter, M. N. & Burns, A. R. Integrin-dependent neutrophil migration in the injured mouse cornea. Exp. Eye Res. 120, 61\u201370, https:\/\/doi.org\/10.1016\/j.exer.2014.01.004 (2014).","journal-title":"Exp. Eye Res."},{"key":"44833_CR16","doi-asserted-by":"publisher","first-page":"6576","DOI":"10.4049\/jimmunol.164.12.6576","volume":"164","author":"XL Rudner","year":"2000","unstructured":"Rudner, X. L., Kernacki, K. A., Barrett, R. P. & Hazlett, L. D. Prolonged elevation of IL-1 in Pseudomonas aeruginosa ocular infection regulates macrophage-inflammatory protein-2 production, polymorphonuclear neutrophil persistence, and corneal perforation. J. Immunol. 164, 6576\u20136582 (2000).","journal-title":"J. Immunol."},{"key":"44833_CR17","doi-asserted-by":"publisher","first-page":"207","DOI":"10.1097\/01.ico.0000248384.16896.7d","volume":"26","author":"G Rohini","year":"2007","unstructured":"Rohini, G., Murugeswari, P., Prajna, N. V., Lalitha, P. & Muthukkaruppan, V. Matrix metalloproteinases (MMP-8, MMP-9) and the tissue inhibitors of metalloproteinases (TIMP-1, TIMP-2) in patients with fungal keratitis. Cornea 26, 207\u2013211, https:\/\/doi.org\/10.1097\/01.ico.0000248384.16896.7d (2007).","journal-title":"Cornea"},{"key":"44833_CR18","doi-asserted-by":"publisher","unstructured":"Chidambaram, J. D. et al. Persistence of Innate Immune Pathways in Late Stage Human Bacterial and Fungal Keratitis: Results from a Comparative Transcriptome Analysis. Frontiers in Cellular and Infection Microbiology 7, https:\/\/doi.org\/10.3389\/fcimb.2017.00193 (2017).","DOI":"10.3389\/fcimb.2017.00193"},{"key":"44833_CR19","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/j.jprot.2014.11.017","volume":"115","author":"RM Selvam","year":"2015","unstructured":"Selvam, R. M. et al. Exoproteome of Aspergillus flavus corneal isolates and saprophytes: identification of proteoforms of an oversecreted alkaline protease. J. Proteomics 115, 23\u201335, https:\/\/doi.org\/10.1016\/j.jprot.2014.11.017 (2015).","journal-title":"J. Proteomics"},{"key":"44833_CR20","doi-asserted-by":"publisher","first-page":"1495","DOI":"10.1016\/j.ophtha.2014.03.004","volume":"121","author":"CQ Sun","year":"2014","unstructured":"Sun, C. Q. et al. Association between in vitro susceptibility to natamycin and voriconazole and clinical outcomes in fungal keratitis. Ophthalmology 121, 1495\u20131500 e1491, https:\/\/doi.org\/10.1016\/j.ophtha.2014.03.004 (2014).","journal-title":"Ophthalmology"},{"key":"44833_CR21","doi-asserted-by":"publisher","first-page":"238","DOI":"10.1097\/00003226-198706040-00002","volume":"6","author":"DM O\u2019Day","year":"1987","unstructured":"O\u2019Day, D. M. Selection of appropriate antifungal therapy. Cornea 6, 238\u2013245 (1987).","journal-title":"Cornea"},{"key":"44833_CR22","doi-asserted-by":"publisher","first-page":"1538","DOI":"10.1016\/S0161-6420(02)01088-6","volume":"109","author":"GK Vemuganti","year":"2002","unstructured":"Vemuganti, G. K. et al. Evaluation of agent and host factors in progression of mycotic keratitis: A histologic and microbiologic study of 167 corneal buttons. Ophthalmology 109, 1538\u20131546 (2002).","journal-title":"Ophthalmology"},{"key":"44833_CR23","doi-asserted-by":"publisher","first-page":"433","DOI":"10.1006\/exer.2000.0971","volume":"72","author":"U Gopinathan","year":"2001","unstructured":"Gopinathan, U. et al. Enzymatic, clinical and histologic evaluation of corneal tissues in experimental fungal keratitis in rabbits. Exp. Eye Res. 72, 433\u2013442, https:\/\/doi.org\/10.1006\/exer.2000.0971 (2001).","journal-title":"Exp. Eye Res."},{"key":"44833_CR24","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1016\/0732-8893(90)90081-6","volume":"13","author":"WS Zhu","year":"1990","unstructured":"Zhu, W. S., Wojdyla, K., Donlon, K., Thomas, P. A. & Eberle, H. I. Extracellular proteases of Aspergillus flavus. Fungal keratitis, proteases, and pathogenesis. Diagn. Microbiol. Infect. Dis. 13, 491\u2013497 (1990).","journal-title":"Diagn. Microbiol. Infect. Dis."},{"issue":"Pt 2","key":"44833_CR25","first-page":"301","volume":"186","author":"JV Jester","year":"1995","unstructured":"Jester, J. V., Petroll, W. M., Barry, P. A. & Cavanagh, H. D. Temporal, 3-dimensional, cellular anatomy of corneal wound tissue. J. Anat. 186(Pt 2), 301\u2013311 (1995).","journal-title":"J. Anat."},{"key":"44833_CR26","doi-asserted-by":"publisher","first-page":"5136","DOI":"10.1167\/iovs.10-7048","volume":"52","author":"A Cruzat","year":"2011","unstructured":"Cruzat, A. et al. Inflammation and the nervous system: the connection in the cornea in patients with infectious keratitis. Invest. Ophthalmol. Vis. Sci. 52, 5136\u20135143, https:\/\/doi.org\/10.1167\/iovs.10-7048 (2011).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"44833_CR27","doi-asserted-by":"publisher","first-page":"1132","DOI":"10.1001\/archopht.121.8.1132","volume":"121","author":"P Hamrah","year":"2003","unstructured":"Hamrah, P., Liu, Y., Zhang, Q. & Dana, M. R. Alterations in corneal stromal dendritic cell phenotype and distribution in inflammation. Arch. Ophthalmol. 121, 1132\u20131140, https:\/\/doi.org\/10.1001\/archopht.121.8.1132 (2003).","journal-title":"Arch. Ophthalmol."},{"key":"44833_CR28","doi-asserted-by":"publisher","first-page":"393","DOI":"10.1007\/s10456-011-9223-3","volume":"14","author":"BB Peebo","year":"2011","unstructured":"Peebo, B. B., Fagerholm, P., Traneus-Rockert, C. & Lagali, N. Cellular level characterization of capillary regression in inflammatory angiogenesis using an in vivo corneal model. Angiogenesis 14, 393\u2013405, https:\/\/doi.org\/10.1007\/s10456-011-9223-3 (2011).","journal-title":"Angiogenesis"},{"key":"44833_CR29","unstructured":"Sperlich, K., Bohn, S., Stolz, H., Guthoff, R. & Stachs, O. In ARVO Imaging in the Eye Conference (Association for Research in Vision and Ophthalmology (ARVO), Hawaii, USA, 2018)."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-44833-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-44833-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-44833-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T10:05:56Z","timestamp":1671271556000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-44833-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,6,6]]},"references-count":29,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["44833"],"URL":"https:\/\/doi.org\/10.1038\/s41598-019-44833-9","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,6,6]]},"assertion":[{"value":"21 September 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 May 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"6 June 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"8334"}}