• Mycobiology
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• 제51권6호
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• pp.452-462
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• 2023

Opportunistic infections due to Cryptococcus neoformans and C. gattii species complexes continue to rise unabated among HIV/AIDS patients, despite improved antifungal therapies. Here, we collected a total of 20 environmental and 25 presumptive clinical cryptococcal isolates from cerebrospinal fluid (CSF) samples of 175 patients enrolled in an ongoing clinical trial Ambition 1 Project (Botswana-Harvard Partnership). Identity confirmation of the isolates was done using MALDI-TOF MS and PCR. We describe the diversity of the isolates by PCR fingerprinting and sequencing (Oxford Nanopore Technology) of the intergenic spacer region. Mating types of the isolates were determined by amplification of the MAT locus. We report an unusual prevalence of 42.1% of C. neoformans × C. deneoformans hybrids Serotype AD (n = 16), followed by 39.5% of C. neoformans Serotype A (n = 15), 5.3% of C. deneoformans, Serotype D (n = 2), 7.9% of C. gattii (n = 3), and 5.3% of C. tetragattii (n = 2) in 38 representative isolates that have been characterized. Mating type-specific PCR performed on 38 representative environmental and clinical isolates revealed that 16 (42.1%) were MATa/MAT𝛼 hybrids, 17 (44.7%) were MAT𝛼, and five (13.2%) possessed MATa mating type. We used conventional and NGS platforms to demonstrate a potential link between environmental and clinical isolates and lay a foundation to further describe mating patterns/history in Botswana.

• Mycobiology
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• 제42권1호
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• pp.52-58
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• 2014

A nucleoside diphosphate-linked moiety X (Nudix) hydrolase-like gene, YSA1, has been identified as one of the gromwell plant extract-responsive genes in Cryptococcus neoformans. Ysa1 is known to control intracellular concentrations of ADP-ribose or O-acetyl-ADP-ribose, and has diverse biological functions, including the response to oxidative stress in the ascomycete yeast, Saccharomyces cerevisiae. In this study, we characterized the role of YSA1 in the stress response and adaptation of the basidiomycete yeast, C. neoformans. We constructed three independent deletion mutants for YSA1, and analyzed their mutant phenotypes. We found that ysa1 mutants did not show increased sensitivity to reactive oxygen species-producing oxidative damage agents, such as hydrogen peroxide and menadione, but exhibited increased sensitivity to diamide, which is a thiol-specific oxidant. Ysa1 was dispensable for the response to most environmental stresses, such as genotoxic, osmotic, and endoplasmic reticulum stress. In conclusion, modulation of YSA1 may regulate the cellular response and adaptation of C. neoformans to certain oxidative stresses and contribute to the evolution of antifungal drug resistance.

• Mycobiology
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• 제42권2호
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• pp.152-157
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• 2014

The iron uptake and utilization pathways play a critical role in allowing human pathogens, including Cryptococcus neoformans, the causative agent of fatal meningoencephalitis, to survive within the mammalian body by competing with the host for iron. Here we show that the iron regulon is also required for diverse environmental stress responses and that in C. neoformans, it is regulated by the high-osmolarity glycerol response (HOG) pathway. Between CFO1 and CFO2, two ferroxidase genes in the iron regulon, CFO1 but not CFO2 was induced during oxidative and osmotic stress. Interestingly, we found that the HOG pathway repressed basal expression of both CFO1 and CFO2. Furthermore, when the HOG pathway was blocked, CFO2 also responded to oxidative and osmotic stress and the response of CFO1 was increased. We also established that CFO1 plays a major role in responding and adapting to diverse environmental stresses, including oxidative and genotoxic damage, osmotic fluctuations, heavy metal stress, and stress induced by cell membrane destabilizers. Therefore, our findings indicate that in C. neoformans, the iron uptake and utilization pathways are not only required for iron acquisition and survival, but also play a significant role in the environmental stress response through crosstalk with the HOG pathway.

• 미생물학회지
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• 제14권4호
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• pp.159-166
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• 1976

This study was carried out for the identification of several fungi isolated from some different geographic regional soil samples and evaluation of its pathgenicity to the mice. isolates, The results were isolated from 200 soil samples and 11 strains from 25 pigeon excreta. 2. Seventeen of Cryptococcus neoformans, eight of Candida albicans and Microporum gypseum, six of M. coorei, and three of Sporothrix schencrii were identified group. 3. An association of C. neoformans with avian habitats was found. Ten of the 11 isolates came from areas frequently by pigeons such as pigeon nest and pigeon excreta. 4. The inflammatory reaction produced in mice that were inoculated with C. neoformans and C. albicans intraperitoneal route.

• 미생물학회지
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• 제42권4호
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• pp.257-264
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• 2006

한국의 임상검체와 자연환경에서 분리된 Cryptococcus neoformans 58주에 대한 혈청형과 세포외효소 생성능에 관한 셍물학적 특성을 조사하였다. 환자로부터 분리된 임상균주 51주 중 48주는 혈청형 A (94.1%) 였으며 2주는 혈청형 B (3.92%),그리고 나머지 1주는 혈청형 D (1.96%)였다. 자연환경에서 분리된 7주는 비둘기 분변에서 분리된 것들이며 모두 혈정형 A였다. 모든 균주는 proteinase와 phnospholipase를 생성하였고, 또한 API-ZYM system을 이용한 19종류의 효소생성능 시험에서는 alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, $\alpha$-glucosidase, 그리고 $\beta$-glucosidase를 생성하였으나, N-acetyl-$\beta$-glucosarninidase는 39주 (67.2%)에서만 생성하였다. 혈청형 B로 동정된 2주와 혈청형 A로 동정된 균주중 1주는$\beta$-glucuronidase를 생성하였다. 본 연구에 사용된 총21종류의 효소 생성능 시험을 기초로 하여 생물형을 구분하였는데, 모두 4가지의 유형을 나타내었고, 또한 임상과 환경균주에서 혈정형과 생물형 특성간의 유의한 상관성를 나타내었다.

• 대한미생물학회지
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• 제21권1호
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• pp.127-131
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• 1986

The mating types and serotypes of 10 clinical isolates of Cryptococcus neoformans have been investigated. Seven isolates were serotype A and three were serotype D and thus they fell in C. neoformans var. neoformans. Mating types of six isolates were found $\alpha$ and two were $\alpha$ but another two isolates were untypable. Enzyme linked immuno-sorbent assay(ELISA) using rabbit hyper-immune serum to cryptococcal polysaccharides was well adapted to the analysis of capsular polysaccharides in sera of the patients with cryptococcal meningitis.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1644-1652
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• 2012

Iron plays a key role in host-pathogen interactions. Microbial pathogens require iron for survival and virulence, whereas mammalian hosts sequester and withhold iron as a means of nutritional immunity. We previously identified two paralogous genes, CFT1 and CFT2, which encode homologs of a fungal iron permease, Cft1 and Cft2, respectively, in the human fungal pathogen Cryptococcus neoformans. Cft1 was shown to play a role in the high-affinity reductive iron uptake system, and was required for transferrin utilization and full virulence in mammalian hosts. However, no role of Cft2 has been suggested yet. Here, we identified the third gene, CFT3, that produces an additional fungal iron permease homolog in C. neoformans, and we also generated the cft3 mutant for functional characterization. We aimed to reveal distinct functions of Cft1, Cft2 and Cft3 by analyzing phenotypes of the mutants lacking CFT1, CFT2 and CFT3, respectively. The endogenous promoter of CFT1, CFT2 and CFT3 was replaced with the inducible GAL7 promoter in the wild-type strain or in the cft1 mutant for gain-of-function analysis. Using these strains, we were able to find that CFT2 is required for growth in low-iron conditions in the absence of CFT1 and that overexpression of CFT2 compensates for deficiency of the cft1 mutant in iron uptake and various cellular stress conditions. However, unlike CFT2, no clear phenotypic characteristic of the cft3 mutant and the strain overexpressing CFT3 was observed. Overall, our data suggested a redundant role of Cft2 in the high-affinity iron uptake and stress responses in C. neoformans.

• 한국미생물·생명공학회지
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• 제38권1호
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• pp.13-18
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• 2010

Past decade systemic mycoses caused by opportunistic human fungal pathogens, including Candida, Aspergillus, and Cryptococcus, have been a growing problem for both immunocompromised and immunocompetent individuals. Particularly, Cryptococcus neoformans has recently emerged as a major fungal pathogen, which can cause fungal pneumonia and meningitis that are lethal if not timely medicated. However, treatment for cryptococcosis has been difficult due to a lack of proper anti-cryptococcal drugs with fungicidal activity and less toxicity. In this review we introduced novel therapeutic methods for treating cryptococcosis by exploring pathogenomic signa1ing networks of C. neoformans with genome-wide transcriptome approaches as well as diverse molecular/genetic tools.

• 미생물학회지
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• 제52권2호
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• pp.226-229
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• 2016

먹물버섯 Coprinellus congregatus의 버섯조직이 자가분해되는 시기에 발현되는 chitinase (Chi2)는 사람에게 질병을 일으키는 Cryptococcus neoformans의 세포벽 두께를 정상 세포벽의 32% 수준까지 감소시킬 뿐만 아니라, 세포 외 섬유상 물질도 제거하였다. Chi2 처리에 의하여 사람에게 알레르기를 유발하는 Alternaria alternata의 생장은 물론 생쥐 모델에서 이 균에 의한 피부알레르기 증상도 감소시켰다.

• 대한간호학회지
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• 제21권1호
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• pp.5-15
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• 1991

The present study was undertaken in an effort to investigate the effects of alcohol on survival of mice and on their humoral and cellular immune responses, The immune responses examined were Arthus and delayed-type hyperrsnesitivity(DTH) reactions to sheep red blood cells(SRBC), contact hypersensitivity to dinitrofluorobenzend(DNFB), antibody response to thymus - dependent SRBC and to thymus -independent polyvinylpyroridone(PVP), and the recovery of Crytococcus neoformans from the liver, spleen, kidney and brain of experimentally infected mice. The administration of ethanol concentrations of 20% or less did not cause any change in survival rates as compared withs saline injected control group. In general, ethanol administration inhibited the Arthus and DTH reactions to SRBC, contact hypersensitivity to DNFB, and antibody response to both SRBC and PVP and it also decreased the resistance of mice to C. neoformans infection. Taken together, the present study stongly suggested that ethanol inhibits immune response and decrease the resistance of mice to C. neoformans infection.