Dissertations / Theses on the topic 'Cryptococcus neoformans'
Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles
Consult the top 50 dissertations / theses for your research on the topic 'Cryptococcus neoformans.'
Next to every source in the list of references, there is an 'Add to bibliography' button. Press on it, and we will generate automatically the bibliographic reference to the chosen work in the citation style you need: APA, MLA, Harvard, Chicago, Vancouver, etc.
You can also download the full text of the academic publication as pdf and read online its abstract whenever available in the metadata.
Browse dissertations / theses on a wide variety of disciplines and organise your bibliography correctly.
Mathoulin-Pelissier, Simone. "Les sérotypes de cryptococcus néoformans en pathologie humaine et dans l'environnement." Bordeaux 2, 1994. http://www.theses.fr/1994BOR23005.
Full textLaurenson, Ian F. "A study of Cryptococcus neoformans varieties gattii and neoformans." Thesis, University of Edinburgh, 1998. http://hdl.handle.net/1842/22395.
Full textSyme, Rachel Mona. "Antigen processing and presentation of Cryptococcus neoformans." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/tape17/PQDD_0014/NQ34704.pdf.
Full textScofield, Melanie. "Heme utilization and storage by Cryptococcus neoformans." Thesis, University of British Columbia, 2009. http://hdl.handle.net/2429/17425.
Full textAlanio, Alexandre. "Dynamique de l'adaptation de Cryptococcus neoformans à l'hôte." Paris 7, 2013. http://www.theses.fr/2013PA077182.
Full textCryptococcosis is an opportunistic infection due to the ubiquitous yeast Cryptococcus neoformans. This pathogen is a facultative intracellular organism. Interaction with immune cells including monocytes/macrophages lineage and dendritic cells are of major importance in the natural history of the infection. In humans, the pathophysiology of the infection evolves in three steps (i) primoinfection in childhood, (ii) dormancy, demonstrated from epidemiological and genotypic data in the lab few years ago (Garcia-Hermoso et al. 1999), (iii) reactivation upon immunosuppression. In terms of disease, clinical presentation and outcome of cryptococcosis are known to be diverse among individuals even those sharing the same underlying diseases. To address the question of the impact of fungal diversity on the natural course of the infection at the macrophage-level (standardized model of yeasts/ j774 macrophages in vitro interaction), murine model-level (murine model of cryptococcosis in OF1 outbred mice) and human-level (CryptoA/D database), we studied (i) the diversity of C. Neoformans/macrophages interactions using well characterized clinical isolates (ii) the correlation between in vitro phenotype of the isolate and clinical outcome in humans (iii) the diversity of adaptation to the host. We developed new assays and new tools using flow cytometry I (quantitative flow cytometry, multispectral imaging flow cytometry, sorting), microscopy (dynamic imaging), gene expression analysis (single-cell quantitative real time PCR) to overcome technical issues. We found high variation in phagocytic, 2 hours-, 48hours-intracellular proliferation indexes among the 54 ClinCn compared to H99. No correlation with the genotype was observed. The lack of sterilization at week 2 despite active antifungal therapy was significantly associated with a lower phagocytic index, whereas treatment failure at month 3 and death from cryptococcosis were significantly related to a higher 2 hours-intracellular proliferation. Among 9 selected clinical isolates compared to H99, (i) the virulence in mice was significantly different, intracellular expression of some virulence factors correlated with (ii) intracellular proliferation and (iii) phagocytic indexes. With a focus on multiplication and stress response and considering the H99 reference strain, we observed the appearance of various populations of yeasts during mice and macrophage infections. After sorting yeasts populations, we observed that a specific one was less prone and dependent of serum to grow compared to the other p'opulations. Gene expression analysis revealed that this population had specific metabolic characteristics that could reflect dormancy. We found a high diversity of C. Neoformans upon interaction with macrophages considering 54 clinical isolates in correlation with clinical outcome in humans, but also a considerable adaptation to host in our two models considering the reference strains H99. We observed also even more diversity of fungal adaptation to host when clinical isolates were considered. Ail together, these data suggest that cryptococcosis and fungal disease in general could be more complex diseases since diversity, plasticity and adaptation of the fungal organism to hosts is high and heterogeneous
Denoyelle, Patricia. "Evaluation comparative du fungitest R avec une méthode de microdilution pour l'étude de la sensibilité aux antifongiques d'isolats de Cryptococcus neoformans." Paris 5, 1999. http://www.theses.fr/1999PA05P056.
Full textVan, de Moer Ariane. "Production et caractérisation d'anticorps monoclonaux anti-Cryptococcus neoformans dirigés contre le galactoxylomannane de la paroi : application au diagnostic et au pronostic des cryptococcoses chez les patients atteints du syndrome de l'immunodéficience acquise." Montpellier 1, 1990. http://www.theses.fr/1990MON13504.
Full textHarrison, Thomas Stephen. "Interactions between Human Immunodeficiency Virus and Cryptococcus neoformans." Thesis, St George's, University of London, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.299059.
Full textOliveira, Fabiana Freire Mendes de. "Análise funcional do gene ERG6 em Cryptococcus neoformans." reponame:Repositório Institucional da UnB, 2013. http://repositorio.unb.br/handle/10482/14252.
Full textSubmitted by Albânia Cézar de Melo (albania@bce.unb.br) on 2013-09-13T15:57:01Z No. of bitstreams: 1 2013_FabianafreireMendesOliveira.pdf: 3550004 bytes, checksum: 479e0f87b52423d1bb6907da07bbdbdc (MD5)
Approved for entry into archive by Marília Freitas(marilia@bce.unb.br) on 2013-10-03T11:53:44Z (GMT) No. of bitstreams: 1 2013_FabianafreireMendesOliveira.pdf: 3550004 bytes, checksum: 479e0f87b52423d1bb6907da07bbdbdc (MD5)
Made available in DSpace on 2013-10-03T11:53:44Z (GMT). No. of bitstreams: 1 2013_FabianafreireMendesOliveira.pdf: 3550004 bytes, checksum: 479e0f87b52423d1bb6907da07bbdbdc (MD5)
O advento de pacientes portadores do HIV e de pacientes imunocomprometidos provocou um aumento na incidência de micoses fúngicas, como a criptococcose. Existem várias classes de antifúngicos disponíveis comercialmente para o tratamento dessas infecções que são utilizadas de acordo com a patologia e o patógeno. No entanto, são relatados casos de resistência a essas drogas e existe uma grande preocupação em relação aos efeitos adversos que elas podem causar nos pacientes. Nesse contexto, o desenvolvimento de novas drogas antifúngicas poderia ser uma solução para a problemática atual das drogas disponíveis e isso requer o estudo de novos alvos moleculares. O gene ERG6 codifica a enzima esterol C-24 metiltransferase que atua na conversão de zimosterol em fecosterol. Constitui uma etapa que ocorre em fungos para a biossíntese do ergosterol, mas não ocorre nos hospedeiros animais, que possuem enzimas específicas para a conversão do zimosterol até colesterol. No presente trabalho, a função do gene ERG6 foi investigada e caracterizada no fungo Cryptococcus neoformans demonstrando que a ausência desse gene gera diversas alterações fenotípicas, tais como sensibilidade ao estresse osmótico, ao estresse oxidativo e maior susceptibilidade a diferentes drogas antifúngicas. Além disso, foi ainda observado que a capacidade de crescer em meios contento diferentes estressores de parede se mostra alterada. Em relação aos fatores de virulência conhecidos para C. neoformans, o fungo foi incapaz de crescer a temperatura de 37°C, porém não teve sua produção de cápsula ou melanina afetadas. No entanto, em testes para observação da sua capacidade de virulência in vitro utilizando macrófagos e in vivo com lagartas da espécie Galleria mellonella mostraram uma redução na virulência em mutantes de ERG6. Por fim a análise dos esteróis de membrana mostrou que ocorre uma alteração em toda a composição de esteróis da membrana celular. Dessa forma, por ser uma enzima encontrada principalmente em fungos, a Erg6 pode ser um alvo molecular potencial para drogas antifúngicas. ______________________________________________________________________________ ABSTRACT
The advent of HIV patients and immunocompromised patients caused an increase in the incidence of fungal mycoses, such as cryptococcosis. There are several antifungal agents commercially available for the treatment of such infections that are chosen based on the disease and the pathogen. However, cases of resistance are reported to these drugs and there is great concern about the adverse effects that they can cause on patients. In this context, the development of new antifungal drugs could be a solution to the current problem of the available drugs and it requires the study of potential molecular targets. The gene ERG6 encodes the sterol C-24 methyltransferase, an enzyme that acts on the conversion zimosterol in fecosterol. This is a step that occurs in fungi in the ergosterol biosynthesis, but does not occur in the animal hosts which have specific enzymes for the conversion of zymosterol to cholesterol. In this study, gene function of ERG6 was investigated and characterized in the Cryptococcus neoformans demonstrating the absence of this gene generates several phenotypic changes, such as sensitivity to osmotic stress, oxidative stress and increased susceptibility to different antifungal drugs. Furthermore, it was also observed that the ability to grow on media with different cell wall stressors was also altered. It was observed that the lack of ERG6 greatly affects the permeabillity of the membrane resulting in osmotic and oxidative stress sensitivity and changing antifungal drugs susceptibility. Furthermore, it was also observed that the ability to grow in medium with cell wall stressor was altered. About the virulence factor, C. neoformans was unable to grow at 37°C, but it had not affected the production of melanin or capsule. However, virulence tests in vitro with macrophages and in vivo with Galleria mellonella caterpillars showed a decrease in the virulence of ERG6 mutants. Finally, the membrane sterols analysis demonstrated a change in the membrane sterol composition. Thus, being an enzyme found especially fungi, the Erg6 may be a potential molecular target for antifungal drugs.
Barros, Amanda Lira Nogueira. "Análise funcional do gene VELB de Cryptococcus neoformans." reponame:Repositório Institucional da UnB, 2014. http://repositorio.unb.br/handle/10482/16480.
Full textSubmitted by Ana Cristina Barbosa da Silva (annabds@hotmail.com) on 2014-10-14T21:29:48Z No. of bitstreams: 1 2014_AmandaLiraNogueiraBarros.pdf: 3370997 bytes, checksum: 85117556d8ae68f1292b6073cbb5fc91 (MD5)
Approved for entry into archive by Guimaraes Jacqueline(jacqueline.guimaraes@bce.unb.br) on 2014-10-15T13:24:45Z (GMT) No. of bitstreams: 1 2014_AmandaLiraNogueiraBarros.pdf: 3370997 bytes, checksum: 85117556d8ae68f1292b6073cbb5fc91 (MD5)
Made available in DSpace on 2014-10-15T13:24:45Z (GMT). No. of bitstreams: 1 2014_AmandaLiraNogueiraBarros.pdf: 3370997 bytes, checksum: 85117556d8ae68f1292b6073cbb5fc91 (MD5)
As proteínas Velvet compreendem quatro membros altamente conservados entre ascomicetos e basidiomicetos, os quais compartilham o domínio Velvet. Eles regulam diferentes vias de sinalização em resposta a estímulos ambientais e também coordenam o metabolismo secundário e diferenciação assexual e sexual em diferentes espécies de fungos. Recentemente, as proteínas Velvet foram implicadas na virulência de alguns agentes patogênicos de plantas. Buscou-se avaliar o papel das proteínas Velvet no fungo patogênico humano Cryptococcus neoformans, causador da criptococose, importante micose sistêmica e a terceira doença mais prevalente em indivíduos portadores do vírus HIV. Entre as micoses sistêmicas, ela é classificada como tendo a maior incidência em pacientes imunocomprometidos. A infecção por C.neoformans ocorre pela inalação de células infecciosas e é considerada uma infecção pulmonar primária, o que pode levar a uma infecção disseminada e meningoencefalite. O objetivo geral do trabalho foi avaliar o papel do gene VELB na patogenidade e morfogênese de C. neoformans. O primeiro passo realizado no estudo foi a deleção do gene VELB em Cryptococcus neoformans. O cassete foi transformado em células leveduriformes haploides de C. neoformans diretamente através de biobalística. Confirmou-se a deleção por PCR e Southern Blot. Após confirmação, os transformantes foram submetidos a diferentes condições de crescimento para determinação de seus fenótipos e virulência. Foram realizados testes de estresse osmótico (NaCl), estresse na parede celular (CongoRed, SDS, cafeína) e estresse oxidativo com peróxido de hidrogênio. Foram avaliados os fatores de virulência de C. neoformans como fosfolipase, urease, melanina e cápsula. Os testes fenotípicos não apresentaram alterações entre a linhagem e mutante indicando que o gene VELB parece não ter nenhum envolvimento em virulência e resposta a agentes estressores. Para avaliação da capacidade de acasalamento foram realizados co-cultivos no escuro em diferentes meios de cultura (Filament, SLAD e MS), além disto foi testada a capacidade de fusão dos mutantes. Estes testes demonstraram que a deleção de VELB bloqueia a produção de hifas no acasalamento entre mutantes e que estes apresentam também defeito no processo de fusão celular. Estes resultados gerados neste trabalho sobre o papel de VELB corroboram os dados anteriormente obtidos pelo nosso grupo de pesquisa de que os membros da família velvet estão diretamente envolvidos na regulação do ciclo sexual de C. neoformans. _________________________________________________________________________________ ABSTRACT
The Velvet proteins consist of four highly conserved members in ascomycetes and basidiomycetes, sharing the Velvet domain. They regulate different signaling ways in response to environmental stimulation and also coordinate secondary metabolism and asexual to sexual differentiation in different species of fungi. Recently, Velvet proteins have been implicated in the virulence of some pathogenic agents of plants. It was evaluated the role of Velvet proteins in the human pathogenic fungus Cryptococcus neoformans that causes cryptococcosis, an important systemic mycosis and the third most prevalent disease in HIV positive individuals. Compared to other systemic mycoses, it is classified as having the highest incidence in immunocompromised patients. C.neoformans infection is by inhalation of infectious cells and is considered a primary pulmonary infection, which can lead to widespread infection and meningoencephalitis. The overall objective of this study was to evaluate the role of the VELB gene in morphogenesis and pathogenicity of C. neoformans. The first step was performed at study deletion of the gene VELB in Cryptococcus neoformans. The cassette was transformed into haploid yeast cells of C. neoformans directly through biolistic. The deletion was confirmed by PCR and Southern blot. After confirmation, the transformants were submitted to different growth conditions to determine their phenotypes and virulence. Tests of osmotic stress (KCl, NaCl, sorbitol), stress in the cell wall (Congo Red, Calcofluor, SDS, caffeine) and oxidative stress with hydrogen peroxide were performed. Virulence factors of C. neoformans as phospholipase, urease, melanin and capsule were evaluated. Phenotypic tests showed no change between the strain and mutant indicating that the VELB gene seems to have no involvement in virulence and response to stressors. To evaluate the ability of mating co-cultures were performed in the dark in different culture media (Filament, SLAD and MS), in addition it was tested the ability of fusion of mutants. These tests showed that the deletion of VELB blocks the production of hyphae in mating and that these mutants also exhibit defective cell fusion process. These results generated in this paper about the VELB function corroborate data previously 13 obtained by our research group that the members of the velvet family are directly involved in the regulation of the sexual cycle of C. neoformans.
Garcia, Hermoso Dea. "Variabilite genotypique chez cryptococcus neoformans : application a l'etude epidemiologique de la cryptococcose (doctorat : microbiologie)." Paris 11, 2001. http://www.theses.fr/2001PA114806.
Full textGoulart, Letícia Silveira. "Genes diferencialmente expressos por Cryptococcus neoformans e Cryptococcus gattii durante a infecção de macrófagos." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2009. http://hdl.handle.net/10183/18796.
Full textAminnejad, Mojgan. "Identification and characterization of novel hybrids within Cryptococcus neoformans and Cryptococcus gattii species complex." Thesis, The University of Sydney, 2013. http://hdl.handle.net/2123/10224.
Full textFoster, Alexander J. "Cell surface analysis of the basidiomycete yeast cryptococcus neoformans." Thesis, Aston University, 2004. http://publications.aston.ac.uk/11011/.
Full textBarcellos, Vanessa de Abreu. "Proteínas bioluminescentes : biomarcadores para o monitoramento in vivo da infecção por Cryptococcus neoformans e Cryptococcus gattii." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2013. http://hdl.handle.net/10183/131973.
Full textCryptococcosis is an invasive fungal disease caused mainly by pathogenic species Cryptococcus neoformans and Cryptococcus gattii. Recent epidemiological studies have suggested that infection with C. gattii develops more frequently pulmonary cryptococcomas, while C. neoformans establishes mainly neurological damages. Although exists this association, the mechanism of dissemination of these species is not completely understood, instigating the use of different approaches for the characterization of the infections. Alternatively, the development of bioluminescent organisms has allowed the real-time monitoring of infection in animal models. In this study, different strategies were used to construct strains of C. neoformans and C. gattii expressing Renilla luciferase reporter gene without success. In a second approach, bioluminescent bacteria (BL) were isolated from seawater samples collected in the vicinity of the river Tramandaí estuarine zone, in order to select the lux cassette (CDABE) for use as a reporter gene. All isolates showed luminescence at 28°C, but when incubated at 37°C, only the isolate BL6 remained with a considerably reduced luminescent activity. Thus, bioluminescent bacteria belonging to the genera Vibrio, Photobacterium and Enterovibrio isolated from Tramandaí river estuary zone presented unsuitable luminescence emission levels at human physiological temperature, preventing the use of lux cassette as a reporter gene in experiments to monitoring cryptococcosis infection.
OLIVEIRA, Patrícia Cariolano de. "Padrão taxonômico, quimiotipagem e atividade antifúngica in vitro de Anfotericina B, Ciclopirox Olamina e β-Lapachona em amostras de Cryptococcus." Universidade Federal de Pernambuco, 2009. https://repositorio.ufpe.br/handle/123456789/1007.
Full textA criptococose é uma infecção fúngica causada por Cryptococcus neoformans, que possui tropismo pelo sistema nervoso central. O propósito deste estudo foi diagnosticar criptococose, identificar variedades de C. neoformans e avaliar a suscetibilidade a anfotericina B, ciclopirox olamina e β-lapachona em amostras da Micoteca URM-UFPE e recém isoladas de Líquor de imunocomprometidos de ambos os sexos atendidos em hospitais de Recife-PE, Brasil. Para o diagnóstico de criptococose, exame direto foi realizado com tinta da China e o semeio em ágar Sabouraud. As variedades foram detectadas com meio L-Canavanina-azul bromotimol. A concentração inibitória seguiu o Clinical and Laboratory Standards Institute. Dezenove casos de criptococose foram diagnosticados com obtenção de 16 culturas. Da Micoteca, foram obtidas 14 amostras de C. neoformans. O exame direto mostrou leveduras capsuladas. C. neoformans var. neoformans ocorreu em 50% dos casos. O CIM (μg/mL) desta variedade foi 0,25 - 4 (anfotericina B), 0,125 - 1 (ciclopirox olamina), 4 e 8 (β-lapachona). Os isolados de C. gattii apresentaram-se 0,25 - 4 (anfotericina B), 0,25-1 (ciclopirox olamina), 4 e 8 (β-lapachona). A concentração fungicida da β-lapachona foi 64 μg/mL. Criptococose predominou no sexo masculino com faixa etária de 25-46 anos e apresentando AIDS como fator de risco predominante. Detectaram-se 4 isolados resistentes a anfotericina B (3 isolados de C. gattii e 1 isolado de C. neoformans var. neoformans). A eficácia do ciclopirox olamina e da β-lapachona, comparada a resistência a anfotericina B, mostra a importância da susceptibilidade associados à variedade para monitorar desenvolvimento de resistência possibilitando descoberta de novas alternativas terapêuticas para criptococose.
Severo, Luiz Carlos. "Criptococose : duas doenças?" reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 1993. http://hdl.handle.net/10183/10275.
Full textSilva, Lívia Kmetzsch Rosa e. "Identificação de genes reguladores pela temperatura na levedura patogênica Cryptococcus neoformans." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2006. http://hdl.handle.net/10183/8555.
Full textSilva, Lívia Kmetzsch Rosa e. "Análise funcional de genes de Cryptococcus neoformans envolvidos no processo de interação patógeno-hospedeiro." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2010. http://hdl.handle.net/10183/132837.
Full textThe expression of pathogen virulence determinants is highly controlled by the host milieu. In this context, the description of crucial genes for hostpathogen interaction is fundamental to better understand the virulence mechanisms utilized during infection. Cryptococcus neoformans is an encapsulated yeast, that causes a life-threatening meningoencephalitis in immunocompromised individuals. The AIDS pandemic has contributed significantly to the increase of scientific research concerning the C. neoformans biology. Functional analyses of three genes, VCX1, GAT1, and GRASP, related to calcium (Ca2+) transport, nitrogen metabolism and unconventional secretion in C. neoformans, respectively, are presented and discussed in this thesis. The ability to survive and proliferate at the human body temperature is an essential virulence attribute of this pathogen. This trait is controlled in part by the Ca2+- calcineurin pathway, which senses and utilizes cytosolic calcium for signaling. C. neoformans VCX1 gene encodes a vacuolar calcium exchanger, which is required for intracellular growth in macrophages and for full virulence in mice. The C. neoformans transcription factor Gat1 regulates the expression of Nitrogen Catabolite Repression (NCR) sensitive genes. Genes involved in ergosterol biosynthesis, iron uptake, cell wall organization and capsule biosynthesis are also Gat1-regulated in C. neoformans. However, Gat1 is not required for survival during macrophage infection and for virulence in a mice model of cryptococcosis. One essential virulence strategy of C. neoformans is the release of glucuronoxylomannan (GXM), a capsular immune-modulatory polysaccharide. Our results demonstrate that GRASP, a golgi reassembly and stacking protein, is involved in GXM secretion, capsule formation and virulence in C. neoformans. Interestingly, the vacuolar calcium exchanger Vcx1 and the transcription factor Gat1 are also involved in GXM secretion, which represents the complexity of this important process in C. neoformans.
PORET, PASCAL. "Meningite a cryptococcus neoformans : aspects actuels a propos d'un cas." Reims, 1988. http://www.theses.fr/1988REIMM042.
Full textTurner, Kylie M. "Identification of residues critical for the function of the fungal virulence factor phospholipase B1 from cryptococcus neoformans." Thesis, The University of Sydney, 2006. https://hdl.handle.net/2123/28063.
Full textAmaro, Maria Cristina de Oliveira. "Caracterização de isolados clínicos de Cryptococcus neoformans e Cryptococcus gattii quanto à susceptibilidade a flunocazol." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2006. http://hdl.handle.net/10183/8038.
Full textIn the last years have occurred an increase of fungal infections due to rise of number of immunocompromised patients, particularly by appearance of the Acquired Immunodeficiency Syndrome (AIDS). The incresing use of antifungal agents has determined the appearance of resistants strains to this agents, already verified with the fluconazole azole, the antifungal agent more used in the maintenance therapy of AIDS patients that developing criptococosis. In this work were determined the Minimuns Inibitories Concentrations (MICs) of fluconazole to Cryptococcus neoformans and Cryptococcus gattii clinical isolates, using the National Commitee for Clinical Laboratory Standards (NCCLS) broth microdilution method. This is the test standard to test antifungal susceptibility to yeasts. Were tested 173 clinical isolates of C. neoformans and C. gattii. The MICs to fluconazole ranged of 0.125 to 16 μg/mL and the MIC50% was 2 and MIC90% was 4μg/mL. All isolates were considered susceptibles to fluconazole. A test of fluconazole resistance inducing was carry out by exposure to drug incresing concentrations of one susceptible isolate, to verify if only the exposure can to result in the resistance development. However occurred only a temporary resistance in one of the subpopulations, since the susceptibility profile was restored. The data obtained in this work are in agreement with national and international studies that also used the standard broth microdilution method. The resistance between C. neoformans and C. gattii clinical isolates has been rare in the world when compared with others yeasts. Nonetheless due to references to resistance development during the maintenance therapy of cryptococosis, it’s important to remain alert as for possibility of appearance and spread of resistants strains.
Andrade, Claudia Sala. "ATIVIDADE DE NOVOS ANTIFÚNGICOS SOBRE Candida spp. E Cryptococcus neoformans." Universidade Federal de Santa Maria, 2004. http://repositorio.ufsm.br/handle/1/5814.
Full textAs infecções fúngicas têm-se tornado uma importante causa de morbidade e mortalidade em pacientes, principalmente os imunocomprometidos. Na tentativa de se transpor esta realidade, têm-se buscado estabelecer e avaliar novas drogas antifúngicas eficazes e com melhor tolerabilidade e segurança. O presente estudo foi objetivado a verificar a atividade dos triazólicos fluconazol, ravuconazol e albaconazol e da micafungina, frente a um banco de fungos leveduriformes incluindo as espécies C.albicans, C. dubliniensis, C. tropicalis, C. lusitaniae, C. glabrata e C. krusei, todas de origem clínica e Cryptococcus neoformans de origens clínica e ambiental. De modo geral, as espécies de Candida spp. e Cryptococcus foram sensíveis aos antifúngicos testados, com exceção da Candida albicans e Candida tropicalis frente ao fluconazol, onde 6,6% e 3,3%, respectivamente dos isolados, foram resistentes a este azólico. Estes resultados demonstram a necessidade de uma constante vigilância da suscetibilidade dos fungos e maiores pesquisas em relação às drogas antifúngicas, pois, desta forma, estaremos contribuindo para a menor morbidade e maior sobrevida dos pacientes.
SILVA, Inês Filipa Horta Pancada Lopes da. "Tipagem molecular de isolados clínicos e ambientais de Cryptococcus neoformans." Master's thesis, Instituto de Higiene e Medicina Tropical, 2010. http://hdl.handle.net/10362/14028.
Full textMycoses are amongst the infectious diseases that are very dangerous affecting all social status in several ways all over the world. These infectious diseases can be superficial, cutaneous, subcutaneous and invasive or systemic, spreading all over the body. A fast and efficient diagnosis of these diseases, including a correct identification of the respective etiological agents, is essential to plan the best treatment to be applied to the patients. An increase in the number of people infected by fungal infections is registered worldwide, as a consequence of an increase in the number of cases of immunocompromised patients and of immunosuppressive therapies, invasive medical procedures and prolonged prescribed treatments, among other aspects. The outcome of the human immunodeficiency epidemics at the beginning of the 80’s, caused by HIV, has also contributed to the increase of the occurrence of opportunistic mycoses. Cryptococcosis is a fungal infection caused by several species of Cryptococcus and presents a worldwide epidemiologic distribution. The most clinically relevant species is Cryptococcus neoformans, whose strains have been traditionally classified into five serotypes related with the antigens of the respective polysaccharide capsules: serotype A (C. neoformans var. grubii), D (C. neoformans var. neoformans); B and C (presently recognised as a distinct but closely related species, C. gattii); and AD (hybrid strains). The main objective of this work was to retrospectively determine the molecular types of a large group of C. neoformans strains that were isolated and maintained for the last 18 years in the Mycology lab of IHMT/UNL. Most of these strains were isolated from immunocompromised patients with Cryptococcosis. The PCR-RFLP of the URA5 gene was used to differentiate the strains. Four molecular types were detected among the isolates: VN1 and VN2 (related to C. neoformans var. grubii, serotype A); VN3 (related with hybrid strains of the serotype AD); and VN4 (related with C. neoformans var. neoformans, serotype D). Molecular types VG1, VG2, VG3 and VG4, related with C. gattii, were not found. VN1 was the most abundant molecular type among clinical isolates (45% of the isolates), followed by VN3 (31%) and VN2 and VN4, both corresponding to 12% of the clinical isolates. The standardized typing method used in this work allowed comparing our results with other recent epidemiological studies on the same subject. In Portugal we have found a greater percentage of C. neoformans molecular types VN1 and VN3 whilst in other European and Latin American countries the most abundant types were 7 VN1 and VN2. Type VN4 was also found to be more abundant in our country. This type seems to be common in Mediterranean areas and is associated to cutaneous manifestations of infections of the central nervous system.
Saavedra, Pedro Henrique Viana. "O papel do inflamassoma na infecção induzida pelo Cryptococcus neoformans." reponame:Repositório Institucional da UnB, 2013. http://repositorio.unb.br/handle/10482/13094.
Full textSubmitted by Albânia Cézar de Melo (albania@bce.unb.br) on 2013-05-13T13:38:11Z No. of bitstreams: 1 2013_PedroHenriqueVianaSaavedra.pdf: 1314251 bytes, checksum: ee5b0094f71f46179c6f13bef2a69329 (MD5)
Approved for entry into archive by Guimaraes Jacqueline(jacqueline.guimaraes@bce.unb.br) on 2013-05-15T11:34:05Z (GMT) No. of bitstreams: 1 2013_PedroHenriqueVianaSaavedra.pdf: 1314251 bytes, checksum: ee5b0094f71f46179c6f13bef2a69329 (MD5)
Made available in DSpace on 2013-05-15T11:34:05Z (GMT). No. of bitstreams: 1 2013_PedroHenriqueVianaSaavedra.pdf: 1314251 bytes, checksum: ee5b0094f71f46179c6f13bef2a69329 (MD5)
Cryptococcus neoformans é um fungo encapsulado patogênico humano que afeta principalmente indivíduos imunossuprimidos. Sua cápsula é um dos mais importantes fatores de virulência, e está envolvida com os processos de evasão da resposta imune e disseminação no hospedeiro. Neste trabalho foi investigado a ativação do inflamassoma por C. neoformans e o papel de sua cápsula nesse processo. Observou-se que o mutante acapsular de C. neoformans induziu elevados níveis da secreção de IL-1β e ativação da protease caspase-1. A secreção de IL-1β foi analisada em macrófagos derivados de medula deficientes para as proteínas ASC, caspase-1, NLRP3 e NLRC4, demonstrando que essa secreção depende do inflamassoma NLRP3, mas não de NLRC4. Além disso, os componentes do inflamassoma se mostraram dispensáveis para fagocitose e atividade antifúngica frente a infecção. Os mecanismos envolvidos no processamento de IL-1β foram dependentes de catepsina B frente ao dano lisossomal e sinalização via quinase Syk. Foi demonstrado também que a sinalização por IL-1β não aumenta a atividade antifúngica de macrófagos. No entanto, ela participa na restrição da taxa de infecção por mecanismos desconhecidos. Estes resultados mostram pela primeira vez que C. neoformans ativa o inflamassoma NLRP3 de forma dependente de caspase-1 através da liberação de catepsina B e sinalização via Syk, levando a produção de IL-1β e restrição da taxa de infecção. Adicionalmente, esses dados demonstram que os isolados encapsulados conseguem diminuir a ativação do inflamassoma. ______________________________________________________________________________ ABSTRACT
Cryptococcus neoformans is an encapsulated human pathogenic fungus that affects primarly immunocompromised individuals. It has a prominent capsule that is an important virulence factor, which is involved in immune response evasion and fungal dissemination. In the present study we investigated whether C. neoformans was able of triggering inflammasome activation and the role of its capsule in this event. The results showed that the acapsular mutant induced high levels of IL-1β secretion and caspase-1 activation. IL-1β -/- -/-secretion induced by C. neoformans was assessed in WT, Asc , Casp1 , -/- -/- Nlrp3 and Nlrc4 bone marrow-derived macrophages, showing IL-1β secretion to be dependent on NLRP3, but independent on NLRC4 inflammasome. In addition, inflammasome components were dispensable for C. neoformans uptake or killing. The mechanisms underlying IL-1β processing and release were dependent on cathepsin B release following lysosomal destabilization and Syk tirosine kinase signaling. We also demonstrate that IL- 1β signaling is not required to restrict intracellular yeast, but limited the rate of infection by an unknown mechanism. Together, our data show for the first time that acapsular C. neoformans activates the NLRP3 inflammasome and IL-1β secretion in a caspase-1 dependent manner and that the mechanisms involved in this event rely on cathepsin B released during lysosomal damage and Syk signaling, leading to restriction of infection rate. Furthermore, our results show that encapsulated C. neoformans is able to dimish inflammasome activation.
Peconick, Luísa Defranco Ferreira. "Caracterização molecular e funcional do gene vosa de Cryptococcus neoformans." reponame:Repositório Institucional da UnB, 2013. http://repositorio.unb.br/handle/10482/14903.
Full textSubmitted by Jaqueline Ferreira de Souza (jaquefs.braz@gmail.com) on 2014-01-02T10:23:34Z No. of bitstreams: 1 2013_LuisaDefrancoFerreiraPeconick_Parcial.pdf: 855251 bytes, checksum: bb95fc3c687f69b402e9466503c903f5 (MD5)
Approved for entry into archive by Marília Freitas(marilia@bce.unb.br) on 2014-01-02T13:07:31Z (GMT) No. of bitstreams: 1 2013_LuisaDefrancoFerreiraPeconick_Parcial.pdf: 855251 bytes, checksum: bb95fc3c687f69b402e9466503c903f5 (MD5)
Made available in DSpace on 2014-01-02T13:07:31Z (GMT). No. of bitstreams: 1 2013_LuisaDefrancoFerreiraPeconick_Parcial.pdf: 855251 bytes, checksum: bb95fc3c687f69b402e9466503c903f5 (MD5)
O basidiomiceto Cryptococcus neoformans, é um fungo oportunista, que comumente infecta pacientes portadores do vírus da imunodeficiência humana (HIV) e imunocomprometidos em geral. É o agente etiológico da criptococose, doença potencialmente fatal e cosmopolita cuja incidência mundial vem se aproximando da de doenças como a tuberculose. O gene VOSA pertence à família velvet, exclusiva de fungos e primeiramente descrita em Aspergillus, e está envolvido na viabilidade dos esporos e na regulação das fases assexuada e sexuada. Proteínas dessa família são fatores transcricionais altamente conservados entre ascomicetos e basidiomicetos que regulam diferentes processos em resposta a estímulos ambientais. A análise in silico do gene VOSA de C. neoformans mostrou sua conservação entre os fungos. Construiu-se um cassete de deleção do gene VOSA contendo o gene de resistência à higromicina B por PCR double joint. O cassete foi transformado por biobalística nas linhagens selvagens KN99a/α (soroA) que permitiu a obtenção de mutantes vosAaΔ e vosAαΔ. Da mesma forma, o fragmento contendo o locus VOSA foi transformado para obtenção do reconstituído tipo a (vosAaΔ::VOSA). Confirmou-se a deleção e a reconstituição de VOSA por PCR comum. Os mutantes vosAaΔ e vosAαΔ foram submetidos aos testes de fenótipo: crescimento a 30°C e 37°C, formação de cápsula, produção de urease e fosfolipase, alterações de parede celular e produção de melanina, no entanto não foram observadas quaisquer alterações no fenótipo quando comparados com a linhagem selvagem. Quanto à capacidade de realizar acasalamento, os mutantes vosAaΔ e vosAαΔ foram submetidos a acasalamento com linhagem selvagem de tipo sexual oposto e apresentaram diminuição na formação de hifas e alteração morfológica significativa de sua estrutura. Mutantes de tipo sexual opostos (vosAaΔ vs. vosAαΔ) também foram submetidos a acasalamento e não apresentaram qualquer formação de hifas. Acredita-se que VOSA seja um regulador positivo da reprodução sexuada, e que controle a formação de estruturas sexuais. A expressão de genes velvet foi analisada no mutante vosAaΔ em comparação à cepa selvagem sem indicação de qualquer alteração transcricional. Os resultados até agora obtidos em C. neoformans colaboram para um melhor entendimento do papel de VOSA na patobiologia deste patógeno. Pretende-se identificar em detalhe o papel deste gene no ciclo de vida de C. neoformans, em especial na produção de esporos, que são as formas infectivas e de disseminação deste patógeno. Os dados moleculares gerados neste trabalho servem como ponto de partida para elucidar a complexa regulação do ciclo sexual de C. neoformans e pela primeira vez descreve o envolvimento de um gene velvet na biologia de um basidiomiceto. ______________________________________________________________________________ ABSTRACT
The basidiomycete fungus Cryptococcus neoformans is an opportunistic organism which commonly infects HIV and immunocompromised patients. It is the etiological agent of cryptococcosis, a potentially fatal and cosmopolitan disease, whose world’s incidence is approaching from diseases such as tuberculosis. VOSA gene belongs to the velvet family, fungal exclusive and primarily described in Aspergillus, involved in the regulation of sexual and asexual development as well as in the viability of the spores. Proteins of this family are transcriptional factors highly conserved among ascomycetes and basidiomycetes regulating different process in response to environmental stimuli. In silico analysis of VOSA gene showed its conservation among the fungi. A VOSA disruption cassette containing the hygromycin reistance gene was assembled by Double Joint PCR. The cassette was transformed by biolistic in KN99a/α (soroA) wild type strains that allowed obtaining the vosAaΔ and vosAαΔ mutants. Likewise, the fragment containing the VOSA locus was transformed to obtain a mating type a reconstituted strain (vosAaΔ::VOSAa). Both disruption and reconstitution were confirmed by traditional PCR. The mutants vosAαΔ and vosAaΔ were tested for various phenotypes: growth at 30 and 37°C, capsule formation, urease and phospholipase production, changes in cell wall and melanin production, however no phenotypic changes were observed compared to the wild type strains. Regarding the ability to mate, vosAaΔ and vosAαΔ mutants were crossed with wild type strain of opposite mating type and the crosses showed decreased hyphae formation and significant morphological alteration of its structure. Mutants of opposite mating type (vosAaΔ vs. vosAαΔ) were subjected to mating and showed no formation of hyphae. VOSA is believed to be a positive regulator of sexual reproduction, and that controlling the formation of sexual structures. Expression of velvet genes was analyzed in vosAaΔ mutant in comparison with wild type strain and any significant transcriptional changes were noticed. The results obtained so far in C. neoformans collaborate for a better understanding of its role in the pathobiology of this pathogen. It is intended to identify in detail the role of VOSA in C. neoformans lyfe cycle, in particular spore production, the infective and dissemination form of the pathogen. The molecular data generated in this study are starting point for a research to elucidate the molecular mechanism of the sexual cycle regulation on C. neoformans and for the first time describ a velvet gene involved in the biology of a basidiomycete.
Clauson, John. "Cryptococcus neoformans Serotype Groups Found in Clinical and Environmental Isolates." TopSCHOLAR®, 1993. http://digitalcommons.wku.edu/theses/1888.
Full textCheng, Po-Yan. "Cryptococcus gattii isolates induce less protective inflammation than Cryptococcus neoformans in a murine model of infection." Thesis, University of British Columbia, 2009. http://hdl.handle.net/2429/7119.
Full textMershon, Kileen Louise. "Complement, passively administered antibodies, and disease dissemination in mice infected with Cryptococcus neoformans or Cryptococcus gattii." Diss., Restricted to subscribing institutions, 2008. http://proquest.umi.com/pqdweb?did=1694502671&sid=1&Fmt=2&clientId=1564&RQT=309&VName=PQD.
Full textSevero, Cecília Bittencourt. "Criptococose em pacientes submetidos a transplante de órgãos sólidos." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2010. http://hdl.handle.net/10183/26147.
Full textIn the period of 1981 to 2010, 54 cases of cryptococcosis in patients with solid organ transplantation indetified at Mycology Laboratory in Santa Casa Hospital Complex, Porto Alegre, RS, were retrospectively studied. Cryptococcois occured in 31 kidney, 13 liver, 7 lung, 2 kidney-pancreas, and 1 heart transplant. The mean age was 47.3 years old (range, 12-76; SD 13.98). A total of 38 patients were male (70.4%). The most frequent clinical manifestation (54 patients) was fever, headache, vomiting, cough and altered mental status. The most common chest radiographic fidings, in 27 patients, were nodules, masses, consolidation, cavitation, and pleural effusion, 10 with proved pulmonary involvement. Thirty four patients had central nervous system involvement, 7 with cutaneous involvement, and 4 at other sites. The cerebospinal fluid, blood, and urine had the highest yield for the microbiologic diagnosis, respectivelly. Nearly all infections in this series of patients with cryptococcosis involved Cryptococcus neoformans (92.7%). By the first time in the literature, we documented C. gattii in lung transplant patients. Finally, considering the type of primary immunosupressive agent used, there was a higher mortality rate on patients with cyclosporine based therapy, and lowest in those with tacrolimus.
Cardoso, Pedro Henrique Magalhães. "Classificação e perfil fenotípico de cepas clínicas e ambientais do complexo Cryptococcus neoformans mantidas em banco de microrganismos." Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/42/42132/tde-12112012-094105/.
Full textAiming to study the phenotypic profile of yeasts maintened in stock culture identified as Cryptococcus neoformans, 40 clinical and 44 environmental strains, were chosen ran domly. The strains had undergone biochemical typing for differentiation as C. neoformans and C. gattii. Among the clinical strains 90% were typed as C. neoformans and 10% as C. gattii, already among the environmental strains were 95,5% C. neoformans and 4,5% C. gattii. The strains thah were positive in biochemical test for C. gattii underwent molecular typing (PCR-RFLP) and found that only four strain were actually C. gattii (VGII) and two other C. neoformans (VNI and VNII). When the factors related to virulence were studied, both the clinical and environmental strains were phospholipase positive but the clinical strains produced a greater amount of this enzyme. All strains were both clinical and environmental production of protease and also showed an intensity colony color (melanization). When the thickness of the capsule was evaluated, all of it showed a capsule, from the clinical strains 67% had an average capsule and from environmental strains 70% had an average capsule. When assessed by sensitivity to antifungal by E-test method all clinical and environmental strains were sensitive to the anphotericine B, ketoconazole, fluconazole, voriconazole and posoconazole. Itraconazole was tested and the samples showed drug sensitive-strains. We point out that phospholipase production would be used as marked to C. neoformans complex and a correct identification of strains maintened in stock culture is necessary.
Bürgel, Pedro Henrique Miranda. "Modulação do inflamassoma por componentes de cápsula secretados pelo Cryptococcus neoformans." reponame:Repositório Institucional da UnB, 2015. http://dx.doi.org/10.26512/2015.02.D.18230.
Full textSubmitted by Albânia Cézar de Melo (albania@bce.unb.br) on 2015-05-20T15:07:31Z No. of bitstreams: 1 2015_PedroHenriqueMirandaBurgel.pdf: 2254644 bytes, checksum: d9741ccd569672b51edd5debc891636f (MD5)
Approved for entry into archive by Guimaraes Jacqueline(jacqueline.guimaraes@bce.unb.br) on 2015-05-21T12:58:45Z (GMT) No. of bitstreams: 1 2015_PedroHenriqueMirandaBurgel.pdf: 2254644 bytes, checksum: d9741ccd569672b51edd5debc891636f (MD5)
Made available in DSpace on 2015-05-21T12:58:45Z (GMT). No. of bitstreams: 1 2015_PedroHenriqueMirandaBurgel.pdf: 2254644 bytes, checksum: d9741ccd569672b51edd5debc891636f (MD5)
O Cryptococcus neoformans é um fungo patogênico humano que afeta majoritariamente indivíduos imunossuprimidos. Ele é o agente causador da meningite criptocócica e tem como principal fator de virulência uma cápsula polissacarídica que recobre sua parede celular. Os componentes desta cápsula estão envolvidos em diversos mecanismos de escape e de supressão da resposta imune desenvolvidos pelo fungo, o que leva a uma resposta não adequada da imunidade inata frente à infecção com consequente disseminação do patógeno. Dentre estes componentes, a glucoronoxilomanana (GXM) se destaca devido à grande variedade de modulações que a mesma é capaz de exercer em diversos tipos celulares. Neste trabalho, demonstramos uma nova interação entre estes componentes capsulares secretados e macrófagos: a inibição da secreção de IL-1β e da piroptose em macrófagos previamente ativados. Primeiramente verificamos que estes componentes possuíam a capacidade de inibir a produção de IL-1β, sem interferir na ativação fornecida pelo primeiro sinal – necessária para a produção da dada citocina. Caracterizamos parcialmente a molécula responsável por esta inibição, chegando a uma molécula pequena (menor que 1kDa), resistente ao calor (121 °C) e polar. Também verificamos que a piroptose dos macrófagos estimulados era prevenida por estes componentes capsulares, confirmando a interferência na via do inflamassoma destas células. Posteriormente realizamos ensaios para definir o mecanismo de ação pelo qual esta interferência estava ocorrendo. Verificamos que existiam depósitos da citocina intracelular pró-IL-1β e também que a enzima caspase-1 estava ativada nos macrófagos em contato com os componentes capsulares do C. neoformans. Por último, realizamos ensaios de interação entre os macrófagos e leveduras de C. neoformans, observando uma replicação intracelular exacerbada do fungo nas células tratadas com os componentes capsulares. Estes resultados demonstram a presença de uma molécula com caráter anti-inflamatório, acrescentando mais um mecanismo de modulação ao já extenso arsenal de fatores de virulência expressos pelo C. neoformans.
Cryptococcus neoformans is a human pathogenic fungus that affects mainly immunocompromised individuals. It is the causative agent of the cryptococcal meningitis and it´s major virulence factor is a polysaccharide capsule that covers it´s cell wall. The components of this capsule are involved in various mechanisms of evasion and suppression of the immune response developed by the fungus, which leads to an inadequate answer of the innate immunity to the infection and consequent dissemination of the pathogen. Among these components, the glucuronoxylomannan (GXM) stands out due to the great variety of modulations that it is able to perform in several cell types. In this study, we demonstrated a novel interaction between these capsular components secreted by the fungus and macrophages: the inhibition of IL-1β secretion and pyroptosis in previously stimulated macrophages. First, we verified that these components had the ability to inhibit the production of IL-1β without interfering in the activation led by the first signal – necessary for the production of this given cytokine. Then we partially characterized the molecule responsible for this inhibition, discovering that it was a small molecule (smaller then 1kDa), heat resistant (121 °C) and polar. We also found that the pyroptosis was being prevented in the stimulated macrophages, confirming the interference in the inflammasome pathway of these cells. Later we performed tests to define the mechanism of action by which these interferences were occurring. We found deposits of the intracellular cytokine pro-IL-1β and also activated caspase-1 enzyme in macrophages that were in contact with C. neoformans capsular components. Ultimately, we performed interaction tests between macrophages and yeast cells of C. neoformans, observing an exacerbated intracellular growth of the fungus inside the cells treated with the capsular components. These results demonstrated the presence of a molecule with anti-inflammatory character, adding another modulatory mechanism into the already extensive arsenal of virulence factors expressed by the C. neoformans.
Mansour, Michael K. "Host response to mannosylated proteins from the pathogenic yeast, Cryptococcus neoformans." Thesis, Boston University, 2005. https://hdl.handle.net/2144/37166.
Full textPLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you.
The pathogenic yeast, Cryptococcus neoformans , is a leading cause of death among individuals with compromised T cell function. Mannoproteins (MP) are a set of functionally heterogeneous, heavily mannosylated glycoforms found in many fungal species including C. neoformans. Cryptococcal MP have been shown to stimulate cell-mediated immunity and pro-inflammatory cytokines, both vital for the clearance of this yeast. As it is unclear how MP elicit immunity, it was hypothesized that the extensive conjugated carbohydrate backbone present on MP plays an essential role in immune stimulation. Mannose receptors (MR), including the macrophage mannose receptor (MMR) and dendritic cell-specific ICAM-3-grabing nonintegrin (DC-SIGN), both present on antigen-presenting cells (APC) and known to recognize mannosylated pathogens were shown to bind MP. Deglycosylation of MP or MR blockade with competitive mannosylated ligands inhibited T-cell activation. The immunodominant APC responsible for immune stimulation was determined by incubating T cells with purified splenic dendritic cells (DC), B cells, and peritoneal macrophages. T cells responded to MP only in the presence of DC. In addition, whole splenocyte populations, but not DC-depleted splenocytes, stimulated MP-specific T cells. Comparing APC populations, only DC captured fluorescent MP in a MR-dependent process. The kinetics of MP capture appear to involve 2 processes, a major, rapid, saturable receptor-mediated process that is inhibited with mannan, and minor pinocytic uptake. Impressively, DC pulsed for short periods with MP were still able to functionally stimulate T cells in a MR-dependent mechanism. Confocal microscopy of MP in DC at early time points indicated co-localization with transferrin, MMR and DC-SIGN suggesting MP enters early endosomes via MR. Subsequently, MP enter degradative perinuclear compartments. In vivo, MP-immunization resulted in increased survival, as well as a decrease in organ fungal load in response to live C. neoformans challenge. This partial protection was dependent on T cells, and not B cells. Moreover, analysis of tissue pro-inflammatory cytokine levels showed an increase in tumor necrosis factor-α and interferon-γ in infected MP-immunized mice. Collectively, these studies begin to provide both a molecular and cellular basis for the immunogenicity of cryptococcal MP and support T cell vaccination strategies that target MR and DCs.
2031-01-01
Pearl, Esther, and n/a. "Requirements for splicing by Cne PRP8, a novel intein from cryptococcus neoformans." University of Otago. Department of Biochemistry, 2006. http://adt.otago.ac.nz./public/adt-NZDU20070405.145703.
Full textNeto, Lauro Vieira PerdigÃo. "Aspectos clÃnicos, epidemiolÃgicos e laboratoriais da criptococose no Estado do Cearà entre os anos de 1985 e 2010, bem como efeitos de antirretrovirais inibidores de protease sobre virulÃncia e crescimento de cryptococcus neoformans in vitro." Universidade Federal do CearÃ, 2011. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=9553.
Full textA criptococose à a infecÃÃo causada a partir da inalaÃÃo de leveduras do gÃnero Cryptococcus. A doenÃa costuma acometer mais freqÃentemente pacientes com alguma imunodepressÃo e evoluir de forma subaguda ou crÃnica. Tem predileÃÃo por sistema nervoso central, manifestando-se como meningite, mas tambÃm pode apresentar-se como doenÃa pulmonar ou de forma disseminada, com envolvimento de outros ÃrgÃos. A atividade proteolÃtica em leveduras do gÃnero Cryptococcus tem sido associada a sua patogenicidade. A atividade da enzima fosfolipase tambÃm se faz importante para este gÃnero, uma vez que està relacionada à nutriÃÃo e à capacidade invasiva do microrganismo. Outro mecanismo de virulÃncia de maior relevÃncia para a patogenicidade de Cryptococcus spp. à a cÃpsula polissacarÃdica presente em sua superfÃcie, responsÃvel principal pelo escape aos mecanismos de defesa do hospedeiro. Esta pesquisa teve como objetivo traÃar um perfil clÃnico, epidemiolÃgico e laboratorial dos pacientes com criptococose entre os anos de 1985 e 2010, bem como investigar os efeitos dos inibidores de protease Saquinavir, Darunavir, Ritonavir e Indinavir sobre o crescimento, espessura da cÃpsula, atividade de protease e expressÃo de fosfolipase em cepas de Cryptococcus neoformans isoladas de humanos. A comparaÃÃo entre grupo soropositivo para o HIV com grupo soronegativo mostrou que o primeiro apresenta maiores mÃdias de idade e maior freqÃÃncia de sexo masculino acometido; por outro lado, o grupo soropositivo demonstra menores proteinorraquia e celularidade no lÃquor, bem como menores valores de leucometria e plaquetometria em sangue perifÃrico. ApÃs a realizaÃÃo do teste de sensibilidade por microdiluiÃÃo em caldo, os fungos foram expostos a trÃs concentraÃÃes distintas dos fÃrmacos e avaliados com relaÃÃo à expressÃo das duas enzimas e espessura da cÃpsula. Os resultados mostraram inibiÃÃo significativa (p<0,05) da atividade de protease pelo menos na maior concentraÃÃo testada para todas as drogas, bem como estreitamento da cÃpsula em algumas combinaÃÃes de drogas e cepas. Por outro lado, quanto à atividade de fosfolipase, observou-se um aumento na expressÃo dessa enzima, especialmente nas concentraÃÃes mais elevadas das quatro drogas. Conclui-se, portanto, que apesar de estes antirretrovirais nÃo inibirem o crescimento de Cryptococcus neoformans, sÃo capazes de alterar mecanismos de virulÃncia destas leveduras.
Gullo, Fernanda Patrícia [UNESP]. "Novas alternativas terapêuticas para o tratamento da Criptococose: análogos de Resveratrol e microRNAs." Universidade Estadual Paulista (UNESP), 2016. http://hdl.handle.net/11449/137768.
Full textRejected by Sandra Manzano de Almeida (smanzano@marilia.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: No campo “Versão a ser disponibilizada online imediatamente” foi informado que seria disponibilizado o texto completo porém no campo “Data para a disponibilização do texto completo” foi informado que o texto completo deverá ser disponibilizado apenas 12 meses após a defesa. Caso opte pela disponibilização do texto completo apenas 12 meses após a defesa selecione no campo “Versão a ser disponibilizada online imediatamente” a opção “Texto parcial”. Esta opção é utilizada caso você tenha planos de publicar seu trabalho em periódicos científicos ou em formato de livro, por exemplo e fará com que apenas as páginas pré-textuais, introdução, considerações e referências sejam disponibilizadas. Se optar por disponibilizar o texto completo de seu trabalho imediatamente selecione no campo “Data para a disponibilização do texto completo” a opção “Não se aplica (texto completo)”. Isso fará com que seu trabalho seja disponibilizado na íntegra no Repositório Institucional UNESP. Por favor, corrija esta informação realizando uma nova submissão. Agradecemos a compreensão. on 2016-03-29T16:52:20Z (GMT)
Submitted by FERNANDA PATRICIA GULLO null (nanda_gullo@hotmail.com) on 2016-04-05T13:11:55Z No. of bitstreams: 1 Tese_Fernanda P. Gullo.pdf: 7327179 bytes, checksum: 0393cbeb07198f30f49ab07572fbb16f (MD5)
Rejected by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: No campo “Versão a ser disponibilizada online imediatamente” foi informado que seria disponibilizado o texto completo porém no campo “Data para a disponibilização do texto completo” foi informado que o texto completo deverá ser disponibilizado apenas 12 meses após a defesa. Caso opte pela disponibilização do texto completo apenas 12 meses após a defesa selecione no campo “Versão a ser disponibilizada online imediatamente” a opção “Texto parcial”. Esta opção é utilizada caso você tenha planos de publicar seu trabalho em periódicos científicos ou em formato de livro, por exemplo e fará com que apenas as páginas pré-textuais, introdução, considerações e referências sejam disponibilizadas. Se optar por disponibilizar o texto completo de seu trabalho imediatamente selecione no campo “Data para a disponibilização do texto completo” a opção “Não se aplica (texto completo)”. Isso fará com que seu trabalho seja disponibilizado na íntegra no Repositório Institucional UNESP. Por favor, corrija esta informação realizando uma nova submissão. Agradecemos a compreensão. on 2016-04-05T13:54:55Z (GMT)
Submitted by FERNANDA PATRICIA GULLO null (nanda_gullo@hotmail.com) on 2016-04-05T15:17:00Z No. of bitstreams: 1 Tese_Fernanda P. Gullo.pdf: 7327179 bytes, checksum: 0393cbeb07198f30f49ab07572fbb16f (MD5)
Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-04-05T17:24:26Z (GMT) No. of bitstreams: 1 gullo_fp_dr_arafc.pdf: 7327179 bytes, checksum: 0393cbeb07198f30f49ab07572fbb16f (MD5)
Made available in DSpace on 2016-04-05T17:24:26Z (GMT). No. of bitstreams: 1 gullo_fp_dr_arafc.pdf: 7327179 bytes, checksum: 0393cbeb07198f30f49ab07572fbb16f (MD5) Previous issue date: 2016-02-29
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação para a Ciência e a Tecnologia (FCT)
Criptococose é uma importante micose sistêmica que acomete principalmente pacientes imunocomprometidos e é classificada como a infecção fúngica com maior mortalidade entre indivíduos portadores de HIV. Cryptococcus neoformans é uma levedura capsulada e o principal agente etiológico da criptococose; encontra-se dispersa no meio ambiente na forma de basidiósporos, os quais são responsáveis pela infecção em humanos e animais. As principais manifestações clínicas estão associadas à infecção pulmonar e meningite. A terapia se dá basicamente com a administração de anfotericina B (AMB) na terapia de ataque associada ou não a 5-fluocitosina e como manutenção é indicado ao longo do tratamento o fluconazol (FCZ). Apesar de esta terapêutica ser eficiente, é constatado elevado número de casos de reincidência e desenvolvimento de resistência aos azóis, além de problemas de toxicidade. Diante desta problemática, este estudo propõe o desenvolvimento de novas alternativas terapêuticas para o tratamento da criptococose, visando duas abordagens distintas. A primeira, aplicando novo composto antifúngico e a segunda, o estudo de microRNAs (miRNAs) envolvidos na interação da levedura e células de glioblastoma humano (U87-MG), com a finalidade de usá-los como reguladores da infecção, sendo esta uma estratégia recentemente divulgada em uma variedade de doenças. Para tanto, inicialmente moléculas análogas de resveratrol foram avaliadas quanto a atividade antifúngica e o derivado orto,orto-HDZ (HDZ) mostrou forte atividade fungicida contra isolados de C. neoformans, com valores de concentração fungicida mínima (CFM) variando entre 0,97 a 7,81 µg/mL. A associação entre HDZ e FCZ mostrou sinergia com potencialização do efeito do FCZ em até 64 vezes para um isolado clínico resistente. Baixa toxicidade foi observada em ensaios in vitro e in vivo (Galleria mellonella e camundongos), apesar de mostrar alterações em embriões de Zebrafish. Além disso, este estudo mostrou a capacidade da associação HDZ + FCZ em inibir a interação da levedura e células hospedeiras (pneumócitos, macrófagos e glioblastoma) por citometria de fluxo e imunofluorescência. Ensaios in vivo (modelo murino) da atividade antifúngica de HDZ em combinação com FCZ mostrou que após 15 dias de tratamento houve redução significativa das unidades formadoras de colônias (UFC) no fígado, baço, cérebro, pulmões e lavado broncoalveolar, assim como o aumento da sobrevida dos animais em 198 dias e redução da morbidade, a qual foi avaliada por ensaio de comportamento (SHIRPA). Diante da segunda proposta deste trabalho, foram encontrados 10 miRNAs super-expressos na interação de C. neoformans e células U87-MG e foi observado que os miRNAs regulam principalmente vias de sinalização TGF-β, MAP quinase (MAPK) e interação receptor e matriz extracelular (MEC). Tais dados foram cruzados com transcritos que mostraram diferencialmente expressos na interação fungo-célula e as proteínas GTPase-3 e COP-1 foram selecionadas como importantes nesse processo. Tais proteínas estão associadas à via Rho-GTPase a qual é primordial para a ligação da levedura em células hospedeiras e mostrou aumento da expressão na infecção, assim como ligeira redução após o tratamento com HDZ. A indução da apoptose de células de glioblastoma por C. neoformans, foi observada através da técnica TUNEL e da expressão da proteína pró-apoptótica Bak e o tratamento com HDZ foi capaz de reverter esse processo. Nossos resultados indicam HDZ como uma interessante molécula para avançar nos estudos de desenvolvimento de um protótipo antifúngico contra C. neoformans e a inibição da expressão dos miRNAs durante a interação fungo-célula, pode ser interessante para controle da infecção fúngica, pois acreditamos que uma vez inibidos, podem reduzir a invasão da levedura em células hospedeiras, sendo então os miRNAs interessantes novos alvos terapêuticos e que o tratamento com HDZ são promissores para o desenvolvimento de um novo antifúngico.
Cryptococcosis is an important systemic mycosis that mainly affects immunocompromised patients and is classified as the fungal infection with higher mortality among individuals with HIV. Cryptococcus neoformans is an encapsulated yeast and the main etiologic agent of cryptococcosis. This yeast is dispersed into the environment in the form of basidiospores, which are responsible for infection in humans and animals. The main clinical manifestations are associated with pulmonary infection and meningitis. The treatment is basically the administration of amphotericin B (AMB) as therapy consolidation with or without the use of 5-flucytosine and, for the maintenance therapy, fluconazole (FCZ) is indicated. Although this therapy is effective, it is observed high number of cases of relapses, development of resistance to azoles and toxicity problems. In front of these problems, this study proposes the development of new therapies for the treatment of cryptococcosis, targeting two distinct approaches. The first aims a new antifungal compound and the second the study microRNAs (miRNAs) involved in the interaction between yeast and human glioblastoma cells (U87-MG), in order to use them as regulators of infection, since this is a recently disclosed strategy in a variety of diseases. For this purpose, initially, analogs of resveratrol molecule were evaluated for antifungal activity and the derived ortho,ortho-HDZ (HDZ) showed a strong fungicidal activity against isolates of C. neoformans with values of minimum fungicidal concentration (MFC) ranging between 0.97 to 7.81 µg/mL. The association between HDZ and FCZ showed synergy with potentiation of the effect of FCZ in up to 64 times for a resistant clinical isolate to FCZ. Low toxicity was observed in in vitro and in vivo assays (Galleria mellonella and mice), despite showing changes in Zebrafish embryos. In vivo assay (murine model) of the antifungal activity of HDZ in combination with FCZ showed that after 15 days of treatment, a significant reduction in colony forming units (CFU) in the liver, spleen, brain, lungs and bronchoalveolar lavage was observed, as well as, increased survival of the animals in 198 days with the reduction of the morbidity, which was evaluated by behavioral assay (SHIRPA). Considering the second purpose of this study, 10 miRNAs were over-expressed during the interaction of C. neoformans and U87-MG cells and it was noted that these miRNAs regulate the signaling pathway of TGF-β, MAP kinase (MAPK) and the receptor and matrix extracellular components (MEC) interaction. These data was cross with transcripts that showed differentially expressed in fungus-cell interaction and the GTPase-3 and COP-1 protein were selected as important in this process. Such proteins are associated to Rho-GT pathway Rho-GTPase which is essential for the yeast adhesion into host cells and showed increased expression in infection, as well as a slight reduction after treatment with HDZ. Induction of apoptosis in glioblastoma cells by C. neoformans was observed by TUNEL technique and the expression of pro-apoptotic protein Bak and treatment with HDZ was able to reverse this process. Our results indicate HDZ as an interesting molecule to advance the development of studies of a prototype antifungal against C. neoformans and that the inhibition of expression of miRNAs during fungus-cell interaction, may be interesting to control fungal infection, because we believe that once inhibited, the yeast can reduce invasion of host cells, being the miRNAs interesting new therapeutic targets and the treatment with HDZ is promising for the development of a new antifungal drug.
CNPq: 403586/2012-7
FCT: 345/13
Paes, Hugo Costa. "Caracterização do módulo regulador do fator de transcrição Mlc1 de Cryptococcus neoformans." reponame:Repositório Institucional da UnB, 2016. http://repositorio.unb.br/handle/10482/22543.
Full textTexto liberado parcialmente pelo autor. Conteúdo restrito: Apêndices.
Submitted by Fernanda Percia França (fernandafranca@bce.unb.br) on 2016-07-20T15:30:33Z No. of bitstreams: 1 2016_HugoCostaPaes_Parcial.pdf: 3794325 bytes, checksum: 3d01a16ba44758f90caa9750abbd4e47 (MD5)
Approved for entry into archive by Raquel Viana(raquelviana@bce.unb.br) on 2017-02-14T21:27:37Z (GMT) No. of bitstreams: 1 2016_HugoCostaPaes_Parcial.pdf: 3794325 bytes, checksum: 3d01a16ba44758f90caa9750abbd4e47 (MD5)
Made available in DSpace on 2017-02-14T21:27:37Z (GMT). No. of bitstreams: 1 2016_HugoCostaPaes_Parcial.pdf: 3794325 bytes, checksum: 3d01a16ba44758f90caa9750abbd4e47 (MD5)
Fatores de transcrição da família Gti1/Pac2 ocorrem exclusivamente em fungos e foram cooptados pela evolução para desempenhar papéis reguladores distintos de acordo com a espécie, papéis esses que vão do controle da transição micélio-levedura e virulência em fungos termodimórficos, à regulação do acasalamento em Candida e do metabolismo secundário em patógenos de plantas. Como essas são funções-chaves para patogênese, levantamos a hipótese de que o homólogo em Cryptococcusneoformans poderia também controlar a virulência, no caso em questão de um importante patógeno fúngico de humanos, que causa uma doença que vitima centenas de milhares de pessoas todo ano. O mutante para o gene que caracterizamos retém a capacidade de secretar diversas substâncias associadas à virulência – melanina, urease e fosfolipase – mas é hipocapsular e tem um defeito de citocinese e de crescimento a 37 °C. É avirulento em camundongos e hipovirulento em Galleria a 30 °C, o que indica que a temperatura do hospedeiro não é a única razão para o fenótipo observado. Além disso, a análise de RNA-Seq mostra que na ausência da proteína correspondente, a expressão da maioria das ORFscodantes do locus MAT é parcial ou completamente reprimida a 37 °C num meio de cultura de células. Entretanto, o mutante não apresenta um defeito na iniciação do acasalamento. Estes resultados dão suporte à idéia de que acasalamento e a resposta a estresses ambientais são mecanismos que evoluíram paralelamente, e é o primeiro relato de um fator de transcrição que controla o locus MAT de um patógeno fúngico num contexto independente do acasalamento. A proteína foi, portanto, nomeada MAT locuscontroller 1 (“controladora do locus MAT 1”, Mlc1).
Gti1/Pac2 transcription factors occur exclusively in fungi and have been co-opted during evolution to perform distinct regulatory roles according to species, ranging from yeast-to-mycelium transition and virulence in dimorphic fungi, to mating in Candida and secondary metabolism in plant pathogens. As these roles are important in causing disease, we hypothesized that the Cryptococcus neoformans homologue could also control virulence in this important fungal pathogen, which causes a disease that kills hundreds of thousands of people each year. The mutant for this gene retains the ability to secrete several substances associated with virulence – melanin, urease and phospholipase – but is hypocapsular and has a cytokinesis and a growth defect at 37 °C. It is avirulent in mice and hypovirulent in Galleria at 30 °C, which indicates that host temperature is not the only reason. Furthermore, RNA-Seq analysis shows that in the absence of the protein, the expression of most protein-coding ORFs of the MAT locus is partial or completely repressed at 37 °C in a cell culture medium. However, the mutant does not have a defect in mating initiation. These results support the idea that mating and the response to environmental stress are co-evolved mechanisms, and is the first report of a transcription factor that controls the MAT locus of a fungal pathogen in a mating-unrelated context. The protein has thus been named MAT locus controller 1 (Mlc1).
Silveira, Carolina Pereira. "Análise do papel da Hsp70 e catalase 2 na fisiologia e patogênese do fungo Cryptococcus neoformans." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2013. http://hdl.handle.net/10183/84966.
Full textCryptococcus neoformans affects immunocompromised patients, especially patients with impaired T cell-mediated immune response, and therefore are susceptible to opportunistic pathogens. C. neoformans has a repertoire of virulence factors that allows the establishment and dissemination of the infection in the central nervous system, causing meningoencephalitis. The virulence factors of Cryptococcus has been subject of many studies that have showed the involvement of gene products in the infection process. However, the treatment of cryptococcal meningoencephalitis consists of antifungal therapy and displays many side effects. For these reasons, new therapies are being addressed, in order to promote the activation of the host immune system to overcome the infection. To develop these therapies, it is necessary to identify molecules that stimulate cell-mediated immune response. Heat shock proteins from many pathogens have been used as a target for the treatment of many humans infectious diseases. These proteins stimulate the T cell-mediated immune response and show promising results in treatment against fungal infections. Similarly, proteins from antioxidant system such as catalase for example, play an important role against free radicals produced by the host and appear to have other functions in the infection process. The understanding of these proteins biology and their involvement with host immune system cells is essential for the validation of its applicability in the treatment of cryptococcosis. In this context, we present here a study of the biological function of a heat shock protein (Hsp70) and one of the enzymes involved in detoxification of reactive oxygen species (catalase 2) of the fungus C. neoformans. We evaluated the therapeutic effects of these proteins in the treatment of cryptococcosis. We have demonstrated that the protein Hsp70 from C. neoformans is located in the polysaccharide capsule, and it is important in the adhesion process of the yeast to epithelial cells A549. Also demonstrated that Cn_rHsp70 co-localizes with the main component of the polysaccharide capsule, glucuronoxilomanana (GXM), on the surface of macrophages. When Cn_rHsp70 are in contact with fungal cells, the cellular viability decreased and does not change the capsule size however, interferes with the GXM secretion. Catalase 2 is deposited on the fungal cell wall and showed activity by hydrogen peroxide degradation. In vivo experiments using animals infected and treated with Hsp70 DNA vaccines and catalase 2, no protection were observed, but there are immune response mediated by Th1 and Th2 cells. The mechanism by which these proteins stimulate the immune system has yet to be elucidated.
Squizani, Eamim Daidrê. "Papel da homeostase de cálcio na patogênese e sinalização celular de Cryptococcus neoformans." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2016. http://hdl.handle.net/10183/158332.
Full textCryptococcus neoformans an opportunistic human pathogen, is the main cause of fungal infections related to death, especially in immunocompromised patients. The infection starts with the colonization of the lung tissue, thereafter the yeast disseminates hematogenously to reach the central nervous system (CNS), causing meningoencephalitis. In order to transmigrate to CNS the fungal cells must cross the blood brain barrier (BBB). For infection establishment, C. neoformans relies on virulence factors, such as polysaccharide capsule, the ability to grow at 37 ºC, and some factors directly related to transmigration. Enzymes as urease, phospholipase B and hyaluronic acid synthase are known as essential components to cross the BBB. The calcium (Ca2+) is a cellular second messenger that participates on calcineurin signaling pathway. This pathway in C. neoformans is crucial and responsible for virulence and host adaptation. Moreover, the regulation of Ca2+ intracellular levels is coordinated by Ca2+ transporters such as Pmc1 and Vcx1, which are the main foccus of our studies. Despite of many advances in understanding the molecular mechanisms of C. neoformans infection the available repertoire of antifungal drugs is still poor, therefore it is necessary to comprise the factors that regulate dissemination to CNS, and also unveil new targets for antifungal therapy. In the present study it was demonstrated through a murine model of systemic infection that the null mutants pmc1 and pmc1vcx1 are avirulent and unable to access the SNC. Besides, the disruption of PMC1 gene leads to alterations on the gene expression profile, mainly in pathways involved with transmigration and calcium homeostasis. Genes related to urease activity, and connected to paracellular mechanism of transmigration had alterations on their expression profile. Taken togheter the result shows the requirement of Pmc1 transporter to transmigration events through BBB and for proper cryptococcosis establishment.
Freitas, Daniel Domingues. "Avaliação in vitro da curcumina frente às cepas de Candida spp. e Cryptococcus neo¬formans resistentes ao fluconazol." reponame:Repositório Institucional da UFC, 2015. http://www.repositorio.ufc.br/handle/riufc/13765.
Full textSubmitted by denise santos (denise.santos@ufc.br) on 2015-10-26T16:23:45Z No. of bitstreams: 1 2015_dis_ddfreitas.pdf: 967080 bytes, checksum: acc93633be99ddf77a8f153781912efc (MD5)
Approved for entry into archive by denise santos(denise.santos@ufc.br) on 2015-10-26T16:32:43Z (GMT) No. of bitstreams: 1 2015_dis_ddfreitas.pdf: 967080 bytes, checksum: acc93633be99ddf77a8f153781912efc (MD5)
Made available in DSpace on 2015-10-26T16:32:43Z (GMT). No. of bitstreams: 1 2015_dis_ddfreitas.pdf: 967080 bytes, checksum: acc93633be99ddf77a8f153781912efc (MD5) Previous issue date: 2015
Considering that some therapeutic properties of curcumin are well described, so the present study points to an antifungal effect of curcumin against strains of pathogenic yeasts resistant to fluconazole, strains of Candida spp. and Cryptococcus neoformans. All strains studied were inhibited by curcumin with different degrees of inhibition among species with MICs ranging from 8-64 mg / ml. After exposure strains of C. albicans, curcumin a decrease in the number of viable cells was observed, thus indicating damage to cell membranes, with possible compromise of their functions. Our results showed that after treatment with curcumin, found in more intense staining with PI (Propidium iodide), indicating changes in cell membranes. In this study, the mitochondrial function of C. albicans cells appears to have been affected after exposure to curcumin. The collapse in Δψm (Mitochondrial Transmembrane Potential) can lead to openings of transient pores in the mitochondrial membranes. In relation to the different mechanisms of action described for curcumin, there is evidence that DNA is one of the celular targets of this molecule. Our data suggest that after exposure to curcumin, cells of C. albicans showed total breaks in DNA strands. In conclusion, the compound curcumin has antifungal activity in vitro against strains of Candida spp. and Cryptococcus neoformans resistant to fluconazole. In addition to promoting DNA damage and externalization of phosphatidylserine, the respective compound seem to act at specific sites near mitochondria of C. albicans cells, leading to death by apoptosis.
Tendo em vista que algumas propriedades terapêuticas da curcumina já são bem descritas, dessa forma o presente estudo aponta para um efeito antifúngico da curcumina frente a cepas de leveduras patogênicas resistentes ao fluconazol, cepas de Candida spp. e Cryptococcus neoformans. Todas as cepas estudadas foram inibidas pela curcumina, com diferentes graus de inibição entre as espécies com CIMs variando entre 8-64 μg/mL. Após a exposição das cepas de C. albicans a curcumina, foi observado uma diminuição no número de células viáveis, indicando assim danos às membranas celulares, com possível comprometimento de suas funções. Os nossos resultados revelaram que após o tratamento com a curcumina, constatamos uma intensificação na marcação com PI (Iodeto de propídio), indicando alterações nas membranas celulares. No presente trabalho, a função mitocondrial das células de C. albicans parece ter sido afetada após exposição à curcumina. O colapso no Δψm (Potencial Transmembrana Mitocondrial) pode conduzir a aberturas de poros transientes nas membranas mitocondriais. Em relação aos diversos mecanismos de ação descritos para a curcumina, existem evidências de que o DNA é um dos alvos celulares desta molécula. Nossos dados sugerem que após a exposição à curcumina, as células de C. albicans, apresentaram quebras totais nas cadeias de DNA. Em conclusão, o composto curcumina apresenta atividade antifúngica in vitro contra cepas de Candida spp. e Cryptococcus neo-formans resistentes ao fluconazol. Além de promover danos ao DNA e externalização da fosfatidilserina, o respectivo composto parecem atuar em sítios específicos próximos a mitocôndria das células de C. albicans, levando à morte por apoptose.
Morales, Bernardina Penarrieta. "Suscetibilidade in vitro de Cryptococcus neoformans e Cryptococcus gattii frente a drogas antifúngicas pela citometria de fluxo." reponame:Repositório Institucional da FIOCRUZ, 2009. https://www.arca.fiocruz.br/handle/icict/8168.
Full textFundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil
Cryptococcus neoformans (C. n) e Cryptococcus gattii (C. g) são agentes da criptococose. A carência de publicações sobre teste de suscetibilidade de C. n e principalmente de C. g é evidente. A maioria de isolados clínicos de C. g mostra-se suscetível in vitro a fluconazol e itraconazol; no entanto o surgimento de resistência a drogas antifúngicas incentivou os testes de suscetibilidade in vitro. Como conseqüência, Clinical and Laboratory Standadards Institute \2013 CLSI, anteriormente National Committee for Clinical Laboratory Standards \2013 NCCLS, publicaram metodologia padronizada M27-A2 para alcançar reprodutibilidade e permitir a comparação de resultados de suscetibilidade entre laboratórios, porém alguns problemas como o tempo de incubação e o padrão de leitura levaram à busca de técnicas alternativas, tais como a citometria de fluxo. O objetivo deste trabalho é padronizar a técnica de citometria de fluxo para o teste de suscetibilidade in vitro de C.n e C.g. visando a redução do tempo de incubação, a utilização de leitura automatizada e a obtenção de resultados rápidos e reprodutíveis A concentração inibitória mínima (CIM) de 20 isolados de C.n e 21 de C.g foi determinada por citometria de fluxo e os resultados comparados com o protocolo padrão proposto pelo CLSI/M27A-2. Após duas horas de incubação com anfotericina B utilizando FUN-1, C. gattii resultou em 100% de concordância entre as duas técnicas para diluição de 2\03BCg/mL e 95,2% para 1\03BCg/mL e C. neoformans resultou em 100% de concordância para 1 e 2\03BCg/mL. Para azólicos e flucitosina, foram obtidos resultados reprodutíveis com o fluorocromo Acridine Orange após 18 horas de incubação, que resultou em 78% de concordância entre as duas técnicas para fluconazol, 85% para itraconazol e 97% para flucitosina. Em ambas as metodologias, C. gattii foi menos suscetível do que C. neoformans frente ao itraconazol e flucitosina (p<0,05). A citometria de fluxo é uma ferramenta útil, com potencial para testes in vitro e determinação da CIM dos antifúngicos estudados, com apreciável redução do tempo mínimo para obtenção de resultados
Cr yptococcus neoformans ( C.n ) and Cryptococcus gattii ( C.g ) are the agents of cryptococcosis. The lack of publications on susceptibility tests of C.n and especially C.g is evident. Most clinical isolates of C.g proved to be susceptible in vitro to fluconazol e and itraconazole and yet the emergence of resistance to antifungal drugs encouraged susceptibility testing in vitro . As a consequence, Clinical and Laboratory Standadards Institute – CLSI ( National Committee for Clinical Laboratory Standards – NCCLS ) pub lished standardized methodology to achieve reproducibility and allow comparison of susceptibility results between laboratories (M27 - A2 document) , however some problems of methodology, like incubation time and standard reading, led to the search for alterna tive techniques, such as flow cytometry. The objective of this work is to standardize the technique of flow cytometry to test the in vitro susceptibility of C . n and C . g in order to reduce the incubation time, use of automated reading and obtain fast and re producible results . The minimum inhibitory concentration (MIC) of 20 isolates of C.n and 21 of C.g was determined by flow cytometry and the results compared with the standard protocol proposed by CLSI/M27A - 2. Flow citometry showed 100% agreement with CLSI/ M27A - 2 results for 2 μ g/mL and 95.2% for 1 μ g/mL dilution when C.g isolates were tested, and 100% agreement for 1 and 2 μ g/ml dilution when C.n were tested after two hours of incubation with amphotericin B using FUN - 1 . Reproducible results were obtained with fluorochrome Acridin e Orange for azoles and flucytosine after 18 hours incubation, resulting in 78% agreement for fluconazole, 85% for itraconazole and 97% for flucytosine. C.g was less susceptible than C.n against itraconazole and flucytosine (p <0.05) in both methodologies. Flow cytometry is a useful tool, with potential for in vitro susceptibility test s of the antifungal agents studied, with appreciable reduction in the minimum time for achieving results.
Mdodo, Rennatus M. "Prevalence and susceptibility of Cryptococcus neoformans to fluconazole in HIV patients in Kenya." Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2010. https://www.mhsl.uab.edu/dt/2010p/mdodo.pdf.
Full textTangen, Kristin Lynne. "Genomic approaches to explore virulence in the fungal pathogen Cryptococcus neoformans." Thesis, University of British Columbia, 2006. http://hdl.handle.net/2429/38338.
Full textScience, Faculty of
Microbiology and Immunology, Department of
Graduate
Gross, Norma Teresa. "Alveolar macrophages and lung surfactant in the defense against Cryptococcus neoformans /." Stockholm, 2000. http://diss.kib.ki.se/2000/91-628-4327-3/.
Full textYan, Zhun Xu Jianping. "Mating system and mitochondrial inheritance in a basidiomycete yeast, Cryptococcus neoformans." *McMaster only, 2006.
Find full textCarroll, Scott. "Functional and genetic dissection of susceptibility to experimental «Cryptococcus neoformans» infection." Thesis, McGill University, 2010. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=95084.
Full textCette thèse étudie les facteurs fonctionnels et génétiques contrôlant la réponse de l'hôte aux infections fongiques causées par la levure Cryptococcus neoformans. Les lignées de souris consanguines C57BL/6J et C3H/HeN sont sensibles à la pneumonie expérimentale à cryptocoques, alors que les lignées CBA/J et SJL/J y sont résistantes. En se basant sur ces observations, trois hypothèses ont été formulées: (1) des différences du système immunitaire inné expliquent la sensibilité différentielle à la pneumonie à cryptocoques des lignées de souris consanguines; (2) la variation naturelle de la sensibilité entre lignées consanguines est un trait génétique complexe sous le contrôle de locus génétiques spécifiques; et (3) la maladie allergique respiratoire est un phénotype commun associé à une sensibilité à la pneumonie à cryptocoques progressive chez les lignées consanguines. La caractérisation fonctionnelle de la réaction immunitaire innée chez les souris C57BL/6J et SJL/J a révélé une réponse pro-inflammatoire accrue chez les souris SJL/J trois heures après l'infection. Cette polarisation a continué tout au long de l'infection, les souris C57BL/6J développant une réponse immunitaire allergique Th2, alors que les souris SJL/J ont présenté une réponse immunitaire Th1. Une analyse in vitro de la signalisation intracellulaire a révélé que la réaction pro-inflammatoire observée chez les souris SJL/J dépend de l'activation prolongée des cascades de signalisation NF-κB et PI3K (phosphatidylinositol 3 kinase). L'étude des liaisons de traits complexes (QTL) chez des souris F2 (C57BL/6J x CBA/J) a révélé un effet du sexe sur le phénotype, justifiant ainsi une analyse séparée. Deux locus de susceptibilité ont été identifiés chez les femelles: Cnes1 (chromosome 1), et Cnes2 (chromosome 17). Chez les mâles, seul le locus Cnes3 (chromosome 17, en aval de Cnes2) a été identifié. Une analyse QTL sur 2 locus a
Evans, Robert J. "The role of Cryptococcus neoformans derived phospholipase B1 during host infection." Thesis, University of Birmingham, 2016. http://etheses.bham.ac.uk//id/eprint/6634/.
Full textSabiiti, Wilber. "Mechanisms of brain infection by the human fungal pathogen Cryptococcus neoformans." Thesis, University of Birmingham, 2012. http://etheses.bham.ac.uk//id/eprint/3861/.
Full textFreitas, Vivianny Aparecida Queiroz. "Ação da curcumina e morina em leveduras do Complexo Cryptococcus neoformans." Universidade Federal de Goiás, 2017. http://repositorio.bc.ufg.br/tede/handle/tede/7402.
Full textApproved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-06-02T11:13:05Z (GMT) No. of bitstreams: 2 Dissertação - Vivianny Aparecida Queiroz Freitas - 2017.pdf: 4680258 bytes, checksum: 0e08c83bdb454e044ddcef0daedb874a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)
Made available in DSpace on 2017-06-02T11:13:05Z (GMT). No. of bitstreams: 2 Dissertação - Vivianny Aparecida Queiroz Freitas - 2017.pdf: 4680258 bytes, checksum: 0e08c83bdb454e044ddcef0daedb874a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-05-12
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
Cryptococcosis is a fungal disease caused by yeasts belonging to the complex Cryptococus neoformans. The antifungal arsenal available for the treatment of this disease is still restricted and is related to the high toxicity and side effects of some drugs, causing great harm to the patients. In this context, it is necessary to discover new bioactives to quell the infections and reduce the adverse effects. The plants have rich source of secondary bioactive metabolites such as tannins, terpenoids, saponins, alkaloids, flavonoids and other compounds, registered with expressive antimicrobial properties. Therefore, the research of natural compounds and derivatives of natural products has been relevant in recent years, due to their relevance in the discovery of new drugs, in addition to which the association of drugs or compounds with different mechanisms of action has been used as an alternative in conventional therapy. We evaluated the mechanisms of action and antifungal effect of natural compounds, curcumin and morin, on Cryptococcus neoformans and Cryptococcus gattii isolates. The antifungal activity was determined across the minimum inhibitory concentration (MIC) and minimum fungicide (MFC). Polyphenols showed MICs ranging from 2 to 256 μg/mL. The compounds showed a synergistic interaction in 30% (6/20) and 15% (3/20) of the isolates when dealing with morin and curcumin, respectively, associated with fluconazole. In the interaction with amphotericin B morin presented synergism in 70% (14/20) of the isolates. Both compounds did not exhibit antagonism in any of the combinations. Changes in fungal cell morphology were observed, and in contact with red blood cells, presented low toxicity. The mechanisms of action revealed that both polyphenols act on the membrane of the fungal cell, inhibiting the synthesis of ergosterol and causing damage to it. These results demonstrate the potential of the compounds as natural bioactives, with great impact on the discovery of new drugs and their efficacy to be used in the treatment of fungal infections.
A Criptococose é uma doença fúngica causada por leveduras pertencentes ao complexo Cryptococus neoformans. O arsenal antifúngico disponível para o tratamento dessa doença ainda é restrito e está relacionado à elevada toxicidade e efeitos colaterais de alguns fármacos, causando grande prejuízo aos pacientes Neste contexto, se faz necessário descobrir novos bioativos para debelar as infecções e reduzir os efeitos adversos. As plantas apresentam rica fonte de metabólitos secundários bioativos como taninos, terpenóides, saponinas, alcalóides, flavonoides e outros compostos, registrados com expressiva propriedade antimicrobiana. Portanto, a pesquisa de compostos naturais e derivados de produtos naturais tem sido pertinente nos últimos anos, devido à sua relevância na descoberta de novos medicamentos, além do que, a associação de fármacos ou compostos, com diferentes mecanismos de ação tem sido utilizada como alternativa na terapia convencional. Nós avaliamos os mecanismos de ação e o efeito antifúngico dos compostos naturais, curcumina e morina, sobre isolados de Cryptococcus neoformans e Cryptococcus gattii. A atividade antifúngica foi averiguada através da determinação da concentração inibitória mínima (CIM) e fungicida mínima (CFM). Os polifenóis, apresentaram CIMs que variaram de 2 a 256 μg/mL. Os compostos apresentaram interação sinérgica em 30% (6/20) e 15%(3/20) dos isolados se tratando da morina e curcumina, respectivamente, associada ao fluconazol. Na interação com a anfotericina B a morina apresentou sinergismo em 70% (14/20) dos isolados. Ambos os compostos não apresentaram antagonismo em nenhuma das combinações. Alterações na morfologia da célula fúngica foram observadas, e em contato com as hemácias, apresentaram baixa toxicidade. Os ensaios de mecanismos de ação revelaram que ambos os polifenóis atuam na membrana da célula fúngica inibindo a síntese de ergosterol e causando danos a mesma. Estes resultados demonstram o potencial dos compostos como bioativos naturais, apresentando grande impacto na descoberta de novos fármacos e sua eficácia para serem utilizados no tratamento de infecções fúngicas.
Silva, Fabiana Brandão Alves. "Aspectos epigenéticos da virulência em Cryptococcus neoformans : papel das histonas desacetilases." reponame:Repositório Institucional da UnB, 2016. http://repositorio.unb.br/handle/10482/21024.
Full textSubmitted by Fernanda Percia França (fernandafranca@bce.unb.br) on 2016-07-01T13:08:28Z No. of bitstreams: 1 2016_FabianaBrandãoAlvesSilva.pdf: 6438172 bytes, checksum: 73b5269f03e7db3bc90d6fea9386fb12 (MD5)
Approved for entry into archive by Raquel Viana(raquelviana@bce.unb.br) on 2016-07-26T22:27:58Z (GMT) No. of bitstreams: 1 2016_FabianaBrandãoAlvesSilva.pdf: 6438172 bytes, checksum: 73b5269f03e7db3bc90d6fea9386fb12 (MD5)
Made available in DSpace on 2016-07-26T22:27:58Z (GMT). No. of bitstreams: 1 2016_FabianaBrandãoAlvesSilva.pdf: 6438172 bytes, checksum: 73b5269f03e7db3bc90d6fea9386fb12 (MD5)
Os genes de histonas desacetilases (HDAC) são altamente conservados entre diferentes espécies e dirigem importantes processos epigenéticos na manutenção da estrutura e funcionamento do genoma. Entretanto, o papel das HDAC na regulação dos traços de virulência de patógenos permanece pobremente explorado. O fungo patogênico em humanos Cryptococcus neoformans passa por mudanças fenotípicas que promovem persistência e sobrevida dentro do hospedeiro ou em nichos ecológicos. Tais mudanças estão associadas à regulação diferencial da expressão gênica. Neste contexto, inicialmente foi avaliado o efeito de dois inibidores químicos de histona desacetilases (HDACi), o butirato de sódio (NaBut) e a tricostatina A (TSA), sobre as células de C. neoformans. Os resultados demonstraram que ambos inibidores foram capazes de afetar os principais traços de virulência de C. neoformans. Em seguida, foram identificados e deletados 8 genes de HDAC de Classe I e Classe II em C. neoformans. A maioria dos processos previamente associados à virulência em C. neoformans foi afetada pelo tratamento com HDACi eou pela deleção de genes: a capacidade de crescimento a 37 – 39 ºC, a formação da cápsula polissacarídica, a produção de melanina, as atividades de fosfolipase e de proteases, a formação de hifas de acasalamento e a integridade da parede celular. Os mutantes de HDAC também mostraram defeitos na sobrevivência intracelular quando co-cultivados com macrófagos murinos, assim como virulência comprometida nos modelos de infecção de Galleria mellonella e camundongos. A histona desacetilase Clr3 foi apontada como um importante regulador da virulência em C. neoformans. A linhagem de C. neoformans clr3mutante mostrou-se hipovirulenta em ambos os modelos de criptococose animal. Outrossim, a análise de RNA-seq indicou que a proteína Clr3 regula a expressão de vários genes importantes para a adaptação e sobrevivência de C. neoformans ao ambiente hospedeiro. Os resultados indicam que a remodelação da cromatina, por meio das conservadas HDAC, é um importante mecanismo envolvido na virulência de C. neoformans. _________________________________________________________________________________________________ ABSTRACT
Histone Deacetylase (HDAC) genes are highly conserved among different species, directing one of the most important epigenetic processes regulating gene expression – chromatin remodeling. However, the role of HDACs in the regulation of virulence traits in pathogens remains poorly explored. The human fungal pathogen Cryptococcus neoformans undergoes phenotypic changes to promote persistence and survival inside the host or in specific ecological niches. Very likely, these changes are associated with epigenetic regulation. In this context, we initially evaluated the effect of two chemical inhibitors of histone deacetylases (HDACi): Sodium butyrate (NaBut) and Trichostatin A (TSA). The results showed that both were able to impair the expression of the main virulence traits of C. neoformans. Based on these data, we identified and deleted eight genes encoding predicted class I/II HDACs in C. neoformans. In this way, we predicted that we would be able to assign specific function to each HDAC, especially in regards to virulence trait expression. Phenotypes of specific HDAC mutant strains indicate that individual proteins control non-identical but overlapping cellular processes associated with virulence. In the other hand, for some genes we also have observed an opposite regulation. Most processes previously related to virulence in C. neoformans were affected here, such as thermotolerance (growth at 37-39 oC); capsule, melanin and protease formation; and cell wall integrity. Additionally, defects in mating and hyphal development were observed for the clr3HDAC mutant strain. HDAC mutants also displayed defects in intracellular survival when co-cultured with activated macrophages, a finding highly correlated with altered virulence in vivo. Also, we tested the virulence in Galleria mellonella and mice. Overall our results support that the Clr3 histone deacetylase is a newly identified regulator of fungal virulence. The corresponding mutant was hypovirulent in both animal models of cryptococcosis. Furthermore transcriptional profiling shows that the Clr3 protein regulates the expression of many genes that are important for the adaptation of C. neoformans for survival inside the host. Our work displays that chromatin remodeling by the conserved histone deacetylases is an important mechanism behind the virulence of C. neoformans.