A 5-Year Retrospective Analysis of the Risk Factors, Treatment, and Prognosis of Patients With Fungal Keratitis in Heilongjiang, China.

PURPOSE: To report the epidemiologic features, laboratory findings, and treatment outcomes of patients with fungal keratitis (FK) during a busy farming period (May, June, and September to November) in Heilongjiang Province, China. DESIGN: Retrospective, observational case series. METHODS: In total, 251 patients diagnosed with FK at the Eye Hospital, First Affiliated Hospital of Harbin Medical University, from 2017 to 2021 were identified. Medical records were retrospectively analyzed, and demographic features, risk factors, monthly distributions, clinical characteristics, laboratory findings, treatment strategies, and prognostic data were collected. RESULTS: The number of FK cases in the busy farming period accounted for 74.1% (186/251) of the total. The mean patient age was 58.2±9.5 years. Males (66.7%) were more likely to develop FK than females (33.3%), and plant-related trauma was the main cause in 80.1% (149/186) of the cases. The most common causative fungal species was Fusarium (34.9%). In vivo confocal microscopy had the highest positivity rate for FK diagnosis (94.6%). The depth of hypopyon, depth of hyphae or spores, and infiltrate width were significantly positively correlated with delayed presentation. CONCLUSIONS: Patients with a longer time of presentation to the hospital were more likely to undergo surgery because of topical medical therapy failure. The most important measures for FK prevention are the use of eye protection during outdoor work and visiting the hospital for treatment as soon as possible after eye injury.

Pyroptosis in pterygium pathogenesis.

Pterygium is a common ocular disease characterized by proliferating fibrovascular tissue. Pyroptosis, a recently discovered programed cell death, is known to be associated with oxidative stress, one of the main causes of pterygia. Here, we aimed to study the role of pyroptosis in pterygium pathogenesis. The expression of nod-like receptor pyrins-3 (NLRP3), caspase-1, IL-18, and IL-1ß was analyzed in 60 human pterygium tissues and 60 human conjunctival epithelium tissues using real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot analysis. Human conjunctival epithelial cells (HConECs) and human pterygium fibroblasts (HPFs) were primary cultured and the level of pyroptosis-associated factors was detected. Both cells were treated with H2O2, and cell lysis was detected by lactate dehydrogenase (LDH) release assay, the expression of the factors by qRT-PCR, Western blot analysis, and immunostaining. The downstream factors IL-18 and IL-1ß were measured after inhibition of caspase-1 to confirm the caspase-1-dependent pyroptosis. α-SMA and E-cadherin were detected as indicators of pyroptosis-induced myofibroblast activation in HPFs. We discovered that the expression of the factors was significantly increased in pterygium and that caspase-1-dependent pyroptosis presents in both H2O2-treated HPFs and HConECs during which the expression of these factors was significantly elevated and the elevation of downstream factors IL-18 and IL-1ß was restrained after caspase-1 inhibition. α-SMA increase and E-cadherin down-regulation were detected in H2O2-treated HPFs and the changes were reversed by caspase-1 inhibition. Pyroptosis displays a role in the pathological process of pterygium formation and progression. Pyroptosis appears to be an intriguing target to prevent pterygium pathogenesis.

Negative roles of a novel nitrogen metabolite repression-related gene, TAR1, in laccase production and nitrate utilization by the basidiomycete Cryptococcus neoformans.

The multicopper oxidase laccase is widespread in fungi and has great industrial importance. One puzzle regarding laccase production in the basidiomycetous yeast Cryptococcus neoformans is that it is inhibited by high temperature (e.g., 37 degrees C). In this paper, we report identification of a nitrogen metabolite repression-related gene, TAR1, which is responsible for laccase repression. Disruption of TAR1 results in a significant increase in the level of LAC1 mRNA at 37 degrees C. The putative protein Tar1 shares a moderate level of similarity with the nitrogen metabolite repressors Nmr1 and NmrA from Neurospora crassa and Aspergillus nidulans, respectively. Likewise, Tar1 has a negative role in the utilization of nitrate. Furthermore, the structure of Tar1 is unique. Tar1 lacks the long C-terminal region of Nmr1 and NmrA. It contains the canonical Rossmann fold motif, GlyXXGlyXXGly, whereas Nmr1 and NmrA have variable residues at the Gly positions. Interestingly, the promoter region of TAR1 contains three TTC/GAA repeats which are likely the heat shock factor (Hsf) binding sites, implying that Hsf has a role in laccase inhibition. TAR1 mediation of temperature-associated repression of LAC1 suggests a novel mechanism of laccase regulation and a new function for Nmr proteins. Our work may be helpful for industry in terms of promotion of laccase activity.

A copper-responsive factor gene CUF1 is required for copper induction of laccase in Cryptococcus neoformans.

The multicopper laccase is a major virulence factor in Cryptococcus neoformans. Its expression regulation is complex. We presented molecular evidence to show that laccase expression was induced by high concentrations of exogenous copper. Melanin production and laccase enzymatic activity increased dramatically in response to the addition of copper to the media. Reverse transcription-PCR amplification of the laccase gene LAC1 mRNA revealed that the induction occurred at the transcriptional level, which required the copper-responsive factor-encoding gene CUF1. Disruption of CUF1 demolished the activation of LAC1 transcription by copper, whereas the reconstituted strain restored the phenotypic defects. Furthermore, copper induction was shown to be independent of derepression by glucose starvation, a well-established activation factor for laccase expression. These results demonstrate a role of the copper-responsive factor gene CUF1 in the expression of laccase in C. neoformans.