• Title/Summary/Keyword: Cryptococcus neoformans

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Sensing the Stress: the Role of the Stress-activated p38/Hog1 MAPK Signalling Pathway in Human Pathogenic Fungus Cryptococcus neoformans

  • Bahn, Yong-Sun;Heitman, Joseph
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2007.05a
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    • pp.120-122
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    • 2007
  • All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.

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Molecular Characterization of Adenylyl Cyclase Complex Proteins Using Versatile Protein-Tagging Plasmid Systems in Cryptococcus neoformans

  • So, Yee-Seul;Yang, Dong-Hoon;Jung, Kwang-Woo;Huh, Won-Ki;Bahn, Yong-Sun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.357-364
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    • 2017
  • In this study, we aimed to generate a series of versatile tagging plasmids that can be used in diverse molecular biological studies of the fungal pathogen Cryptococcus neoformans. We constructed 12 plasmids that can be used to tag a protein of interest with a GFP, mCherry, $4{\times}FLAG$, or $6{\times}HA$, along with nourseothricin-, neomycin-, or hygromycin-resistant selection markers. Using this tagging plasmid set, we explored the adenylyl cyclase complex (ACC), consisting of adenylyl cyclase (Cac1) and its associated protein Aca1, in the cAMP-signaling pathway, which is critical for the pathogenicity of C. neoformans. We found that Cac1-mCherry and Aca1-GFP were mainly colocalized as punctate forms in the cell membrane and non-nuclear cellular organelles. We also demonstrated that Cac1 and Aca1 interacted in vivo by co-immunoprecipitation, using $Cac1-6{\times}HA$ and $Aca1-4{\times}FLAG$ tagging strains. Bimolecular fluorescence complementation further confirmed the in vivo interaction of Cac1 and Aca1 in live cells. Finally, protein pull-down experiments using $aca1{\Delta}$::ACA1-GFP and $aca1{\Delta}$::ACA1-GFP $cac1{\Delta}$ strains and comparative mass spectrometry analysis identified Cac1 and a number of other novel ACC-interacting proteins. Thus, this versatile tagging plasmid system will facilitate diverse mechanistic studies in C. neoformans and further our understanding of its biology.

Antifungal Mechanism of Action of Lauryl Betaine Against Skin-Associated Fungus Malassezia restricta

  • Do, Eunsoo;Lee, Hyun Gee;Park, Minji;Cho, Yong-Joon;Kim, Dong Hyeun;Park, Se-Ho;Eun, Daekyung;Park, Taehun;An, Susun;Jung, Won Hee
    • Mycobiology
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    • v.47 no.2
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    • pp.242-249
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    • 2019
  • Betaine derivatives are considered major ingredients of shampoos and are commonly used as antistatic and viscosity-increasing agents. Several studies have also suggested that betaine derivatives can be used as antimicrobial agents. However, the antifungal activity and mechanism of action of betaine derivatives have not yet been fully understood. In this study, we investigated the antifungal activity of six betaine derivatives against Malassezia restricta, which is the most frequently isolated fungus from the human skin and is implicated in the development of dandruff. We found that, among the six betaine derivatives, lauryl betaine showed the most potent antifungal activity. The mechanism of action of lauryl betaine was studied mainly using another phylogenetically close model fungal organism, Cryptococcus neoformans, because of a lack of available genetic manipulation and functional genomics tools for M. restricta. Our genome-wide reverse genetic screening method using the C. neoformans gene deletion mutant library showed that the mutants with mutations in genes for cell membrane synthesis and integrity, particularly ergosterol synthesis, are highly sensitive to lauryl betaine. Furthermore, transcriptome changes in both C. neoformans and M. restricta cells grown in the presence of lauryl betaine were analyzed and the results indicated that the compound mainly affected cell membrane synthesis, particularly ergosterol synthesis. Overall, our data demonstrated that lauryl betaine influences ergosterol synthesis in C. neoformans and that the compound exerts a similar mechanism of action on M. restricta.

Comparison of Virulence between Five Strains of Cryptococcus Species Complex in a Rat Model

  • Park, Gyu-Nam;Kim, Sun-Young;Kim, Hye-Ran;Jung, Bo-Kyung;An, Dong-Jun;Hong, Seung-Bok;Chang, Kyung-Soo
    • Biomedical Science Letters
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    • v.24 no.3
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    • pp.183-195
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    • 2018
  • Cryptococcosis, which is caused by the Cryptococcus species complex (including Cryptococcus neoformans and Cryptococcus gattii), is well known as one of the most important medical problems. However, the of the Cryptococcus species complex is still limited to pneumonia and meningitis. In particular, the differences in virulence among the five major serotypes of the Cryptococcus species complex are not fully understood. To elucidate the virulence of the Cryptococcus species complex when it is disseminated hematogenously, rats were infected by different strains of the Cryptococcus species serotype, and their histopathological characteristics were compared after infection. The cumulative mortality ratio of rats infected with serotype B strain was slightly higher than in the other experimental groups. In addition, the average recovery of the Cryptococcus species complex from rats infected with serotype B strain was significantly higher than in the other groups in almost all organ samples except spleen. The recovery of the Cryptococcus species complex was associated with the severity of histopathological lesions, including bleeding, inflammation, and tissue damage in all organs. In rats infected with serotype B strain, the virulence was the most severe, especially in the lungs and liver. These results indicate that the pathophysiology of the Cryptococcus species complex infection differs according to serotype.

Studies on the Distribution of Several Pathogenic Fungi and Its Pathogenicity Isolated from Avian Excreta and Soil (조류분 및 토양에서 분리된 수종 병원성 진균의 검색 및 병원성에 관항 연구)

  • Koh, Choon-Myung;Lew, Joon
    • Korean Journal of Microbiology
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    • v.14 no.4
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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.

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Antifungal Activity of Essential Oil from Asarum sieboldii against Epidermal and Opportunistic Pathogenic Fungi (세신 정유 추출물의 피부진균 및 기회감염진균에 대한 항진균 효과)

  • Han, Kap-Hoon
    • The Korean Journal of Mycology
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    • v.35 no.1
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    • pp.58-60
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    • 2007
  • Root of Asarum sieboldii has been used in chinese medicine. It was been known that the extract of A. sieboldii has medicinal effect such as antifebrile, anodyne, and anti-inflammatory. Antifungal activity of the essential oil extracted from A. sieboldii against various epiderman and opportunistic human pathogenic fungi was investigated. Steam-distillated essential oil demonstrated very high antifungal activity against Aspergillus fumigatus, Aspergillus niger, Cryptococcus neoformans, and Candida albicans. MIC (minimum inhibitory concentration) of the essential oil against C. albicans was 5 ul/ml, indicating that the essential oil may be used as natural preserves for cosmetic and nutritional products.

Microbe Hunting: A Curious Case of Cryptococcus

  • Bartlett, Karen H.;Kidd, Sarah;Duncan, Colleen;Chow, Yat;Bach, Paxton;Mak, Sunny;MacDougall, Laura;Fyfe, Murray
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2005.06a
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    • pp.45-72
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    • 2005
  • C. neoformans-associated cryptococcosis is primarily a disease of immunocompromised persons, has a world-wide distribution, and is often spread by pigeons in the urban environment. In contrast, C. gattii causes infection in normal hosts, has only been described in tropical and semi-tropical areas of the world, and has a unique niche in river gum Eucalyptus trees. Cryptococcosis is acquired through inhalation of the yeast propagules from the environment. C. gattii has been identified as the cause of an emerging infectious disease centered on Vancouver Island, British Columbia, Canada. No cases of C. gattii-disease were diagnosed prior to 1999; the current incidence rate is 36 cases per million population. A search was initiated in 2001 to find the ecological niche of this basidiomycetous yeast. C. gaftii was found in the environment in treed areas of Vancouver Island. The highest percentage of colonized-tree clusters were found around central Vancouver Island, with decreasing rates of colonization to the north and south. Climate, soil and vegetation cover of this area, called the Coastal Douglas fir biogeoclimatic zone, is unique to British Columbia and Canada. The concentration of airborne C. gattii was highest in the dry summer months, and lowest during late fall, winter, and early spring, months which have heavy rainfall. The study of the emerging colonization of this organism and subsequent cases of environmentally acquired disease will be informative in planning public health management of new routes of exposure to exotic agents in areas impacted by changing climate and land use patterns. Cryptococcosis is an infection associated with an encapsulated, basidiomycetous yeast Cryptococcus neoformans. The route of entry for this organism is through the lungs, with possible systemic spread via the circulatory system to the brain and meninges. There are four cryptococcal serogroups associated with disease in humans and animals, distinguished by capsular polysaccharide antigens. Cryptococcus neoformans: variety grubii (serotype A), variety neoformans (serotype D), and variety gattii (serotypes B and C) (Franzot et at. 1999). C. neoformans variety gattii has recently been elevated to species status, C. gattii. C. neoformans val. grubii and var. neoformans have a world-wide distribution, and are particularly associated with soil and weathered bird droppings. In contrast, C. gattii (CG) is not associated with bird excrement, is primarily found in tropical and subtropical climates, and has a restricted environmental niche associated with specific tree species. (Ellis & Pfiffer 1990) Ellis and Pfeiffer theorize that, as a basidiomycete, CG requires an association with a tree in order to become pathogenic to mammals. In Australia, CG has been found to be associated with five species of Eucalypts, Eucalyptus camaldulensis, E. tereticornis, E. blakelyi, E. gomphocephala, and E. rudis. Eucalypts, although originally native to Australia, now have a world-wide distribution. CG has been found associated with imported eucalypts in India, California, Brazil, and Egypt. In addition, in Brazil and Columbia, where eucalypts have been naturalized, native trees have been shown to harbour CG (Callejas et al. 1998; Montenegro et al. 2000). In British Columbia, Canada, since the beginning of 1999, there have been 120 confirmed cases of cryptococcal mycoses associated with CG in humans, including 4 fatalities (data from British Columbia Centre for Disease Control), and over 200 cases in animal pets in BC (data from Central Laboratory for Veterinarians). What is remarkable about the BC outbreak of C. gattii-cryptococcosis is that all of the cases have been residents of, or visitors to, a narrow area along the eastern coast of Vancouver Island, BC, from the tip of the island in the south (Victoria) to Courtenay on the north-central island as illustrated in Figure 1. Of the first 38 human cases, 58% were male with a mean age of 59.7 years (range 20 - 82): 36 cases (95%) were Caucasian. Ten cases (26%) presented with meningitis, the remainder presented with respiratory symptoms. Cultures recovered from cases of cryptococcosis associated with the outbreak were typed as serogroup B, which is specific to CG (Bartlett et al. 2003). This was the first reported outbreak of CVG in Canada, or indeed, the world. Where infection with CG is endemic, for example, Australia, the incidence of cryptococcosis ranges from 1.8 - 4.7 per million between the southern and northern states (Sorrell 2001). However, the overall incidence of cryptococcosis in immunocompenent individuals has been estimated at 0.2 per million population per year (Kwon-Chung et al. 1984). The population of Vancouver Island is approximately 720,000,consequently, even if the organism were endemic, one would expect a maximum of 0.15 cases of cryptococcal disease annually.

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Cryptococcal Granuloma (Cryptococcal Granuloma -1예 보고-)

  • 심성보
    • Journal of Chest Surgery
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    • v.19 no.2
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    • pp.341-346
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    • 1986
  • Crytococcosis is a subacute or chronic, very rare in incidence and very often fatal infection caused by Cryptococcus Neoformans. The organism is of world-wide distribution, affecting both sexes and all ages. The most common portals of entry appear to be respiratory tract. Recently we experienced one case of isolated Cryptococcal Granuloma on the right lower lobe. The patient was 63 year old housewife, who complained only of epigastric pain. On baseline studies of chest X-ray, we incidentally found the abnormal finding of mass density on right lower lobe.

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