Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica

Detalhes bibliográficos
Autor(a) principal: Midoricava, Luana Rossi Oliveira [UNESP]
Data de Publicação: 2015
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://hdl.handle.net/11449/134141
Resumo: Histoplasma capsulatum var. capsulatum is a dimorphic fungi that causes Histoplasmosis, a systemical fungal disease that is a major health problem worldwide. Infection occurs by inhalation of infective propagules from the environment. The biofilm formation by this fungi was characterized recently, however little is known about the mechanisms involved in their formation. In this case, studies with proteomic approaches would be of fundamental importance for understanding the difference between the planktonic and biofilm forms. The antifungal agents used in conventional therapy are not effective against biofilms, in addition, high doses of these drugs can be toxic to patients. This study aimed to verify the anti-Histoplasma activity of 93 compounds derived from chalcones and protocatechuic acid; characterize the biofilm formation by the clinical isolate H. capsulatum 317 and discover new anti-biofilm compounds searching for a mechanism of action by differential proteomics analysis, comparing the treated and untreated biofilms. Susceptibility tests were performed according to the document M27- A3, proposed by the CLSI (2008). The biofilms were formed in BHI broth supplemented and metabolic activity was determined by the XTT reduction assay. For the characterization of biofilm formation and action of the compound, was used the scanning electron microscopy (SEM) and confocal microscopy, while the mass of the biofilm and the extracellular matrix were quantified by crystal violet and safranin. Lastly was performed the analysis of the protein profile of biofilms with and without treatment. The best minimum inhibitory concentration values (MIC) were obtained for six derivatives of chalcones (T50, C3, E2A, F2A, T18A and T46A) and nonyl protocatechuate (MIC = 3.9 mg/L). The clinical isolate H. capsulatum EH317 was able to form biofilms in vitro after 96 hours. After 96 hours, the biofilm ...
id UNSP_95efafcfedb0d50450ae22956f99679a
oai_identifier_str oai:repositorio.unesp.br:11449/134141
network_acronym_str UNSP
network_name_str Repositório Institucional da UNESP
repository_id_str 2946
spelling Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômicaHistoplasmoseFungoBiofilmeMicologiaProteômicaHistoplasmosisHistoplasma capsulatum var. capsulatum is a dimorphic fungi that causes Histoplasmosis, a systemical fungal disease that is a major health problem worldwide. Infection occurs by inhalation of infective propagules from the environment. The biofilm formation by this fungi was characterized recently, however little is known about the mechanisms involved in their formation. In this case, studies with proteomic approaches would be of fundamental importance for understanding the difference between the planktonic and biofilm forms. The antifungal agents used in conventional therapy are not effective against biofilms, in addition, high doses of these drugs can be toxic to patients. This study aimed to verify the anti-Histoplasma activity of 93 compounds derived from chalcones and protocatechuic acid; characterize the biofilm formation by the clinical isolate H. capsulatum 317 and discover new anti-biofilm compounds searching for a mechanism of action by differential proteomics analysis, comparing the treated and untreated biofilms. Susceptibility tests were performed according to the document M27- A3, proposed by the CLSI (2008). The biofilms were formed in BHI broth supplemented and metabolic activity was determined by the XTT reduction assay. For the characterization of biofilm formation and action of the compound, was used the scanning electron microscopy (SEM) and confocal microscopy, while the mass of the biofilm and the extracellular matrix were quantified by crystal violet and safranin. Lastly was performed the analysis of the protein profile of biofilms with and without treatment. The best minimum inhibitory concentration values (MIC) were obtained for six derivatives of chalcones (T50, C3, E2A, F2A, T18A and T46A) and nonyl protocatechuate (MIC = 3.9 mg/L). The clinical isolate H. capsulatum EH317 was able to form biofilms in vitro after 96 hours. After 96 hours, the biofilm ...Histoplasma capsulatum variedade capsulatum é um fungo dimórfico causador da Histoplasmose, uma doença fúngica sistêmica que constitui um importante problema de saúde mundial. A infecção ocorre através da inalação de propágulos infectantes provenientes do meio ambiente. A capacidade de formação de biofilme por esse fungo foi caracterizada recentemente, porém ainda pouco se sabe sobre os mecanismos envolvidos na sua formação. Neste caso, estudos com abordagens proteômicas seriam de fundamental importância para o entendimento da diferença entre a forma planctônica e o biofilme. Os antifúngicos usados na terapia convencional contra biofilmes não são eficientes, além disso, doses elevadas para perfusão destes fármacos nesta forma seriam tóxicas para os pacientes. O presente trabalho teve como objetivo verificar a atividade anti-Histoplasma de 92 compostos derivados de chalcona e do ácido protocatecuico; caracterizar o biofilme do isolado clínico H. capsulatum 317 e pesquisar compostos anti-biofilme buscando um provável mecanismo de ação através da análise proteômica diferencial entre o biofilme tratado e não tratado. Os testes de susceptibilidade foram realizados conforme o documento M27-A3, proposto pelo CLSI (2008). Os biofilmes foram formados em caldo BHI suplementado e a atividade metabólica foi determinada através do ensaio de redução do XTT. Para a caracterização da formação do biofilme e da ação do composto, utilizou-se a microscopia eletrônica de varredura (MEV) e microscopia confocal, enquanto que a massa do biofilme e a matriz extracelular foram quantificadas pelo cristal violeta e safranina. E como etapa final foi realizada a análise do perfil proteico dos biofilmes com e sem tratamento. Os melhores valores de concentração inibitória mínima (CIM) foram obtidos por seis derivados de chalconas (T50, C3, E2A, F2A, T18A e T46A) e...Universidade Estadual Paulista (Unesp)Almeida, Ana Marisa Fusco [UNESP]Universidade Estadual Paulista (Unesp)Midoricava, Luana Rossi Oliveira [UNESP]2016-02-05T18:29:55Z2016-02-05T18:29:55Z2015-08-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis122 f. : figs., tabsapplication/pdfMIDORICAVA, Luana Rossi Oliveira. Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica. 2015. 122 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Ciências Farmacêuticas, 2015.http://hdl.handle.net/11449/134141000854060000854060.pdf33004030081P7Alephreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPporinfo:eu-repo/semantics/openAccess2024-06-24T18:10:08Zoai:repositorio.unesp.br:11449/134141Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T20:02:01.817632Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica
title Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica
spellingShingle Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica
Midoricava, Luana Rossi Oliveira [UNESP]
Histoplasmose
Fungo
Biofilme
Micologia
Proteômica
Histoplasmosis
title_short Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica
title_full Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica
title_fullStr Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica
title_full_unstemmed Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica
title_sort Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica
author Midoricava, Luana Rossi Oliveira [UNESP]
author_facet Midoricava, Luana Rossi Oliveira [UNESP]
author_role author
dc.contributor.none.fl_str_mv Almeida, Ana Marisa Fusco [UNESP]
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Midoricava, Luana Rossi Oliveira [UNESP]
dc.subject.por.fl_str_mv Histoplasmose
Fungo
Biofilme
Micologia
Proteômica
Histoplasmosis
topic Histoplasmose
Fungo
Biofilme
Micologia
Proteômica
Histoplasmosis
description Histoplasma capsulatum var. capsulatum is a dimorphic fungi that causes Histoplasmosis, a systemical fungal disease that is a major health problem worldwide. Infection occurs by inhalation of infective propagules from the environment. The biofilm formation by this fungi was characterized recently, however little is known about the mechanisms involved in their formation. In this case, studies with proteomic approaches would be of fundamental importance for understanding the difference between the planktonic and biofilm forms. The antifungal agents used in conventional therapy are not effective against biofilms, in addition, high doses of these drugs can be toxic to patients. This study aimed to verify the anti-Histoplasma activity of 93 compounds derived from chalcones and protocatechuic acid; characterize the biofilm formation by the clinical isolate H. capsulatum 317 and discover new anti-biofilm compounds searching for a mechanism of action by differential proteomics analysis, comparing the treated and untreated biofilms. Susceptibility tests were performed according to the document M27- A3, proposed by the CLSI (2008). The biofilms were formed in BHI broth supplemented and metabolic activity was determined by the XTT reduction assay. For the characterization of biofilm formation and action of the compound, was used the scanning electron microscopy (SEM) and confocal microscopy, while the mass of the biofilm and the extracellular matrix were quantified by crystal violet and safranin. Lastly was performed the analysis of the protein profile of biofilms with and without treatment. The best minimum inhibitory concentration values (MIC) were obtained for six derivatives of chalcones (T50, C3, E2A, F2A, T18A and T46A) and nonyl protocatechuate (MIC = 3.9 mg/L). The clinical isolate H. capsulatum EH317 was able to form biofilms in vitro after 96 hours. After 96 hours, the biofilm ...
publishDate 2015
dc.date.none.fl_str_mv 2015-08-28
2016-02-05T18:29:55Z
2016-02-05T18:29:55Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv MIDORICAVA, Luana Rossi Oliveira. Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica. 2015. 122 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Ciências Farmacêuticas, 2015.
http://hdl.handle.net/11449/134141
000854060
000854060.pdf
33004030081P7
identifier_str_mv MIDORICAVA, Luana Rossi Oliveira. Prospecção de substâncias anti histoplasma capsulatum nas formas planctônica e biofilme e análise proteômica. 2015. 122 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Ciências Farmacêuticas, 2015.
000854060
000854060.pdf
33004030081P7
url http://hdl.handle.net/11449/134141
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 122 f. : figs., tabs
application/pdf
dc.publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.source.none.fl_str_mv Aleph
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
_version_ 1808129152598933504