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1.
J. appl. oral sci ; 25(3): 274-281, May-June 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-893617

RESUMO

Abstract Pulpal and periodontal tissues have similar microbiota that allows cross-contamination between the pulp and periodontal tissues. Objective The aim of this study was to investigate the prevalence of isolated Candida albicans from periodontal endodontic lesions in diabetic and normoglycemic patients, and the fungi's virulence in different atmospheric conditions. Material and Methods A case-control study was conducted on 15 patients with type 2 diabetes mellitus (G1) and 15 non-diabetics (G2) with periodontal endodontic lesions. Samples of root canals and periodontal pockets were plated on CHROMagar for later identification by polymerase chain reaction (PCR) and virulence test. Results C. albicans was identified in 79.2% and 20.8% of the 60 samples collected from diabetic and normoglycemic patients, respectively. Of the 30 samples collected from periodontal pockets, 13 showed a positive culture for C. albicans, with 77% belonging to G1 and 23% to G2. Of the 11 positive samples from root canals, 82% were from G1 and 18% from G2. Production of proteinase presented a precipitation zone Pz<0.63 of 100% in G1 and 72% in G2, in redox and negative (Pz=1), under anaerobic conditions in both groups. Hydrophobicity of the strains from G1 indicated 16.4% with low, 19.3% with moderate, and 64.3% with high hydrophobicity in redox. In G2, 42.2% had low, 39.8% had moderate, 18% had high hydrophobicity in redox. In anaerobic conditions, G1 showed 15.2% with low, 12.8% with moderate, and 72% with high hydrophobicity; in G2, 33.6% had low, 28.8% had moderate, and 37.6% had high hydrophobicity. There was statistical difference in the number of positive cultures between G1 and G2 (p<0.05) with predominance in G1. There was statistical difference for all virulence factors, except hemolysis (p=0.001). Conclusions Candida albicans was isolated more frequently and had higher virulence in diabetic patients.


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Doenças Periodontais/microbiologia , Candida albicans/isolamento & purificação , Candida albicans/patogenicidade , Doenças da Polpa Dentária/microbiologia , Diabetes Mellitus Tipo 2/microbiologia , Oxirredução , Peptídeo Hidrolases/análise , Doenças Periodontais/fisiopatologia , Doenças Periodontais/diagnóstico por imagem , Bolsa Periodontal/microbiologia , Fosfolipases/análise , Virulência , DNA Fúngico , Radiografia Dentária , Estudos de Casos e Controles , Reação em Cadeia da Polimerase , Estatísticas não Paramétricas , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/fisiopatologia , Doenças da Polpa Dentária/diagnóstico por imagem , Diabetes Mellitus Tipo 2/fisiopatologia , Eletroforese , Interações Hidrofóbicas e Hidrofílicas
2.
Braz. j. microbiol ; 47(4): 911-916, Oct.-Dec. 2016. tab
Artigo em Inglês | LILACS | ID: biblio-828186

RESUMO

Abstract Objective Candida albicans is the primary causative agent of oral candidosis, and one of its key virulent attributes is considered to be its ability to produce extracellular phospholipases that facilitate cellular invasion. Oral candidosis can be treated with polyenes, and azoles, and the more recently introduced echinocandins. However, once administered, the intraoral concentration of these drugs tend to be sub-therapeutic and rather transient due to factors such as the diluent effect of saliva and cleansing effect of the oral musculature. Hence, intra-orally, the pathogenic yeasts may undergo a brief exposure to antifungal drugs. We, therefore, evaluated the phospholipase production of oral C. albicans isolates following brief exposure to sub-therapeutic concentrations of the foregoing antifungals. Materials and methods Fifty C. albicans oral isolates obtained from smokers, diabetics, asthmatics using steroid inhalers, partial denture wearers and healthy individuals were exposed to sub-therapeutic concentrations of nystatin, amphotericin B, caspofungin, ketoconazole and fluconazole for one hour. Thereafter the drugs were removed and the phospholipase production was determined by a plate assay using an egg yolk-agar medium. Results The phospholipase production of these isolates was significantly suppressed with a percentage reduction of 10.65, 12.14, 11.45 and 6.40% following exposure to nystatin, amphotericin B, caspofungin and ketoconazole, respectively. This suppression was not significant following exposure to fluconazole. Conclusions Despite the sub-therapeutic, intra oral, bioavailability of polyenes, echinocandins and ketoconazole, they are likely to produce a persistent antifungal effect by suppressing phospholipase production, which is a key virulent attribute of this common pathogenic yeast.


Assuntos
Humanos , Fosfolipases/biossíntese , Candida albicans/efeitos dos fármacos , Candida albicans/metabolismo , Candidíase Bucal/microbiologia , Candidíase Bucal/tratamento farmacológico , Antifúngicos/farmacologia , Polienos/uso terapêutico , Polienos/farmacologia , Azóis/uso terapêutico , Azóis/farmacologia , Candida albicans/isolamento & purificação , Candida albicans/patogenicidade , Fumar , Testes de Sensibilidade Microbiana , Dentaduras , Fatores de Virulência , Diabetes Mellitus , Ativação Enzimática , Espaço Extracelular , Equinocandinas/farmacologia , Antifúngicos/uso terapêutico
3.
Bol. micol. (Valparaiso En linea) ; 30(2): 6-15, dic. 2015. ilus, tab
Artigo em Espanhol | LILACS | ID: biblio-868801

RESUMO

Introducción: la criptocococis es una micosis sistémica causada por C. neoformans y C. gattii, es frecuente y oportunista en inmunocomprometidos y patógeno primario en personas inmunocompetentes. C. neoformans tiene una distribución mundial y se ha aislado desde las excretas de palomas. C. gattii se considera restringida a regiones con clima tropical, subtropical, y templadas, se encuentra asociada frecuentemente a detritos de especies de Eucalyptus sp. La virulencia de estas levaduras le permite desarrollar patogénesis en mamíferos y supervivencia en el ambiente. Objetivo: Identificar y determinar la actividad de proteinasas y fosfolipasas, de C. neoformans y C. gattii aisladas desde las oquedades de árboles en lugares con alta afluencia de público. Materiales y Métodos: Se tomaron 200 muestras de hisopado desde distintas especies de árboles desde sectores de la región de O’Higgins y el Maule. Se siembran en ASG, se aíslan y mantienen en ASD. Identificación con tinta china, Urea de Christensen, crecimiento a 37°C, asimilación y fermentación de azucares, y siembra en medio CGB. Se mide índice de actividad enzimática Prz de proteinasas y fofolipasas. Resultados y Conclusiones: Se obtuvieron 109 cepas de C. neoformans aisladas desde las oquedades de diferentes especies arbóreas y 3 cepas presuntivas de C. gattii desde Eucalyptus sp. y Prunus cerasifera artropurpurea. El 88,1 por ciento de las cepas C. neoformans y 100 por ciento de C. gattii, presentaron alta actividad proteolítica, El 49,5 por ciento de las cepas de C. neoformans y 33,3 por ciento de C. gattii mostraron alta actividad de fosfolipasas.


Introduction: criptocococis is a systemic mycosis caused by C. neoformans and C. gattii, frequent and opportunistic in immunocompromised and primary pathogen in immunocompetent persons. C. neoformans has a worldwide distribution and has been isolated from the excreta of pigeons. C. gattii is considered restricted to regions with tropical, subtropical, and temperate, is often associated with species of Eucalyptus sp. The virulence of these yeasts develop pathogenesis allows survival in mammals and the environment. Objective: To identify and determine the activity of proteinases and phospholipases of C. neoformans and C. gattii isolated from the hollows of trees in places with high turnout. Materials and Methods: 200 swab samples were taken from different species of trees from areas of the region of O’Higgins and Maule. Planted in ASG, they are isolated and kept in ASD. Identification with ink, Urea Christensen, growth at 37 ° C, assimilation and fermentation of sugars, and planting medium CGB. Prz index proteinase enzyme activity is measured and phospholipases. Results and Conclusions: We manage to get 109 strains of C. neoformans isolated from the hollows of different tree species and 3 presumptive strains of C. gattii from Eucalyptus sp. and Prunus cerasifera artropurpurea. 88.1 percent of the strains C. neoformans and C. gattii 100 percent , they showed high proteolytic activity, 49.5 percent of the strains of C. neoformans and C. gattii 33.3 percent showed high activity phospholipases.


Assuntos
Humanos , Cryptococcus gattii/isolamento & purificação , Cryptococcus gattii/enzimologia , Cryptococcus gattii/patogenicidade , Cryptococcus neoformans/isolamento & purificação , Cryptococcus neoformans/enzimologia , Cryptococcus neoformans/patogenicidade , Peptídeo Hidrolases , Fosfolipases , Árvores/microbiologia , Chile , Criptococose/etiologia , Eucalyptus/microbiologia , Pneumopatias Fúngicas , Prunus/microbiologia
4.
Rev. bras. enferm ; 67(6): 920-927, Nov-Dec/2014.
Artigo em Português | LILACS, BDENF - Enfermagem | ID: lil-732823

RESUMO

Este estudo objetivou compreender as práticas de cuidado dos profissionais de saúde que assistem os idosos Kaingang. Estudo qualitativo, apoiado na etnografia, realizado com dez profissionais à que atuam na atenção primária saúde da Terra Indígena Faxinal, Paraná, Brasil. Os dados foram coletados no período de novembro de 2010 a fevereiro de 2012 por meio da observação participante e entrevistas, e, analisados à luz da Teoria Transcultural do Cuidado. Identificaram-se como práticas de cuidado a medicação e imunização, bem como, cuidados da medicina tradicional. Para realização destes cuidados, os profissionais dispunham de estratégias que proporcionavam manutenção dos idosos na assistência. Conclui-se que valores culturais e científicos necessitam integrar a assistência para melhoria da saúde dos idosos indígenas.


This research aims to understand the care practices of health professionals who assist the elderly Kaingang. It is a qualitative study, supported in ethnography, conducted by ten professionals working in primary health care in the indigenous land of Faxinal, Paraná, Brazil. The data was collected from November 2010 to February 2012 by participant observation and interviews, and analyzed based on the Transcultural Care Theory. Was identified the preoccupation of the carers practices with the medication and immunization, as well as traditional medical care. To achieve these, care professionals had strategies that implemented maintenance of older people in care. We conclude that cultural values and integrate scientific need assistance to improve the health of elderly indigenous.


Este estudio tuvo como objetivo entender las prácticas de cuidado de los profesionales de la salud que asisten a los ancianos Kaingang. Estudio cualitativo, apoyado en la etnografía, llevado a cabo con diez profesionales que trabajan en la atención primaria de la salud de la tierra indígena de Faxinal, Paraná, Brasil. Los datos fueron recogidos a partir de noviembre 2010 a febrero 2012 a través de la observación participante y las entrevistas, y analizado con base en la Teoría del Cuidado Transcultural. Se identificaron las prácticas de atención médica y imunizacion,el cuidado de la medicina, así tradicional. Para lograrlo, los profesionales tenían estrategias que proporcionaban el mantenimiento de las personas mayores en su atención. Se concluye que los valores culturales y científicos necesitan ayuda para mejorar la salud de los ancianos indígenas.


Assuntos
Animais , Ratos , Fígado/enzimologia , Lisossomos/enzimologia , Fosfolipases A/metabolismo , Fosfolipases/metabolismo , Inibidores de Proteases/farmacologia , Células Cultivadas , Quimotripsina/antagonistas & inibidores , Inibidores de Cisteína Proteinase/farmacologia , Leucina/análogos & derivados , Leucina/farmacologia , Leupeptinas/farmacologia , Oligopeptídeos/farmacologia , Pepstatinas/farmacologia , Fosfolipases A1 , Fatores de Tempo
5.
Braz. j. oral sci ; 12(3): 199-204, July-Sept. 2013. graf, tab
Artigo em Inglês | LILACS | ID: lil-701306

RESUMO

An essential factor to the virulence of the genus Candida is the ability to produce enzymes and this may be crucial in the establishment of fungal infections. AIM:This study investigated in vitro enzymatic activities of Candida species and their virulence in an in vivo Galleria mellonella experimental model. METHODS: Twenty-four clinical strains of Candida spp. isolated from the human oral cavity were evaluated, including the following species: C. albicans, C. dubliniensis, C. glabrata, C. tropicalis, C. krusei, C. parapsilosis, C. norvegensis, C. lusitaniae and C. guilliermondii. All Candida strains were tested in vitro for production of proteinase and phospholipase. The Candida strains were also injected into Galleria mellonella larvae to induce experimental candidiasis, and after 24 hours, the survival rate was assessed. RESULTS: Phospholipase and proteinase activity were observed in 100% of the C. albicans strains. In the non-albicans species, proteinase and phospholipase activity were observed in 25 and 43% of the studied strains, respectively. The most pathogenic Candida species in G. mellonella were C. albicans, C. dubliniensis and C. lusitaniae, whereas C. glabrata was the least virulent species. Furthermore, a positive significant correlation was found between both enzymatic activities with virulence in G. mellonella. CONCLUSIONS: The virulence of Candida strains in G. mellonella is related to the quantity of proteinases and phospholipases production of each strain.


Assuntos
Humanos , Candida/patogenicidade , Invertebrados/patogenicidade , Peptídeo Hidrolases , Fosfolipases , Fatores de Virulência
6.
Rev. Soc. Bras. Med. Trop ; 46(3): 340-342, May-Jun/2013. tab
Artigo em Inglês | LILACS | ID: lil-679510

RESUMO

Introduction Candida albicans is a commensal and opportunistic agent that causes infection in immunocompromised individuals. Several attributes contribute to the virulence and pathogenicity of this yeast, including the production of germ tubes (GTs) and extracellular hydrolytic enzymes, particularly phospholipase and proteinase. This study aimed to investigate GT production and phospholipase and proteinase activities in bloodstream isolates of C. albicans. Methods One hundred fifty-three C. albicans isolates were obtained from blood samples and analyzed for GT, phospholipase, and proteinase production. The assays were performed in duplicate in egg yolk medium containing bovine serum albumin and human serum. Results Detectable amounts of proteinase were produced by 97% of the isolates, and 78% of the isolates produced phospholipase. GTs were produced by 95% of the isolates. A majority of the isolates exhibited low levels of phospholipase production and high levels of proteinase production. Conclusions Bloodstream isolates of C. albicans produce virulence factors such as GT and hydrolytic enzymes that enable them to cause infection under favorable conditions. .


Assuntos
Animais , Bovinos , Humanos , Candida albicans/enzimologia , Candida albicans/crescimento & desenvolvimento , Peptídeo Hidrolases/biossíntese , Fosfolipases/biossíntese , Fatores de Virulência/biossíntese , Candida albicans/patogenicidade , Soroalbumina Bovina
7.
Artigo em Inglês | LILACS | ID: lil-686621

RESUMO

Backgound: The venom of the Cuban scorpion Rhopalurus junceus is poorly study from the point of view of their components at molecular level and the functions associated. The purpose of this article was to conduct a proteomic analysis of venom components from scorpions collected in different geographical areas of the country. Results: Venom from the blue scorpion, as it is called, was collected separately from specimens of five distinct Cuban towns (Moa, La Poa, Limonar, El Chote and Farallones) of the Nipe-Sagua-Baracoa mountain massif and fractionated by high performance liquid chromatography (HPLC); the molecular masses of each fraction were ascertained by mass spectrometry analysis. At least 153 different molecular mass components were identified among the five samples analyzed. Molecular masses varied from 466 to 19755 Da. Scorpion HPLC profiles differed among these different geographical locations and the predominant molecular masses of their components. The most evident differences are in the relative concentration of the venom components. The most abundant components presented molecular weights around 4 kDa, known to be K+-channel specific peptides, and 7 kDa, known to be Na+-channel specific peptides, but with small molecular weight differences. Approximately 30 peptides found in venom samples from the different geographical areas are identical, supporting the idea that they all probably belong to the same species, with some interpopulational variations. Differences were also found in the presence of phospholipase, found in venoms from the Poa area (molecular weights on the order of 14 to 19 kDa). The only ubiquitous enzyme identified in the venoms from all five localities studied (hyaluronidase) presented the same 45 kD molecular mass, identified by gel electrophoresis analysis. Conclusions: The venom of these scorpions from different geographical areas seem to be simila, and are rich in peptides that have of the same molecular masses of the peptides...


Assuntos
Animais , Peptídeos , Fosfolipases , Proteômica , Venenos de Escorpião/isolamento & purificação , Cuba/epidemiologia , Espectrometria de Massas/métodos
8.
Braz. j. med. biol. res ; 45(5): 411-416, May 2012. ilus, tab
Artigo em Inglês | LILACS | ID: lil-622763

RESUMO

Fusarium species have emerged as one of the more outstanding groups of clinically important filamentous fungi, causing localized and life-threatening invasive infections with high morbidity and mortality. The ability to produce different types of hydrolytic enzymes is thought to be an important virulence mechanism of fungal pathogens and could be associated with the environment of the microorganism. Here, we have measured the production of two distinct lipolytic enzymes, phospholipase and esterase, by sixteen Fusarium isolates recovered from the hospital environment, immunocompromised patients’ blood cultures, foot interdigital space scrapings from immunocompromised patients, and foot interdigital space scrapings from immunocompetent patients (4 isolates each). Fourteen of these 16 isolates were identified asFusarium solani species complex (FSSC) and two were identified as F. oxysporum species complex (FOSC). Some relevant genus characteristics were visualized by light and electron microscopy such as curved and multicelled macroconidia with 3 or 4 septa, microconidia, phialides, and abundant chlamydospores. All Fusarium isolates were able to produce esterase and phospholipase under the experimental conditions. However, a negative correlation was observed between these two enzymes, indicating that a Fusarium isolate with high phospholipase activity has low esterase activity and vice versa. In addition, Fusarium isolated from clinical material produced more phospholipases, while environmental strains produced more esterases. These observations may be correlated with the different types of substrates that these fungi need to degrade during their nutrition processes.


Assuntos
Humanos , Esterases/biossíntese , Fusarium/enzimologia , Fosfolipases/biossíntese , Fusarium/patogenicidade , Fusarium/ultraestrutura , Microscopia Eletrônica de Varredura , Especificidade da Espécie
9.
Rev. Inst. Med. Trop. Säo Paulo ; 54(1): 17-24, Jan.-Feb. 2012. tab
Artigo em Inglês | LILACS, Sec. Est. Saúde SP | ID: lil-614891

RESUMO

INTRODUCTION: In HIV-infected patients, colonization of the oral cavity by potential pathogenic yeast may lead to development of systemic fungemia. We evaluated the prevalence of yeast in the oral cavity of Brazilian HIV-positive patients and verified whether or not the species characterized were enzymatically active. Furthermore, the species identified were tested for their susceptibility to antifungal treatment. METHODS: Patient saliva and oropharyngeal candidiasis samples were collected from 60 seropositive HIV patients and identified by the API20C system. Enzymatic activity was evaluated by the production of proteinase and phospholipase. Susceptibility to antifungal treatments were determined using the broth microdilution method. RESULTS: the most commonly isolated species were C. albicans (51.56 percent) followed by non-albicans Candida species (43.73 percent), Trichosporon mucoides (3.12 percent) and Kodamaea ohmeri (1.56 percent). Oral colonization by association of different species was observed in 42 percent of the patients. Enzymatic activity was verified in most of species isolated, except for C. glabrata, C. lusitaniae and C. guilliermondii. Resistance to Fluconazole and Amphotericin B was observed in isolates of C. albicans, C. glabrata, C. parapsilosis, C. krusei, and K. ohmeri. CONCLUSION: HIV-positive patients are orally colonized by single or multiple species of yeast that are occasionally resistant to Fluconazole or Amphotericin B.


INTRODUÇÃO: Em pacientes infectados pelo HIV, a colonização da cavidade bucal por leveduras patogênicas pode levar ao desenvolvimento de fungemias. No presente estudo, avaliamos a prevalência de leveduras na cavidade bucal de pacientes HIV-positivos e verificamos se as espécies isoladas foram enzimaticamente ativas. Além disso, as espécies identificadas foram testadas quanto à suscetibilidade a antifúngicos. MÉTODOS: Amostras de saliva e de candidose orofaríngea foram coletadas de 60 pacientes soropositivos para HIV e identificados pelo sistema API20C. A atividade enzimática foi avaliada pela produção de proteinase e fosfolipase. A suscetibilidade a antifúngicos foi determinada utilizando o método de microdiluição em caldo. RESULTADOS: As espécies mais comumente isoladas foram C. albicans (51,56 por cento), seguido por espécies de Candida não-albicans (43,73 por cento), Trichosporon mucoides (3,12 por cento) e Kodamaea ohmeri (1,56 por cento). A colonização bucal por associação de diferentes espécies foi observada em 42 por cento dos pacientes. A atividade enzimática foi verificada na maioria das espécies isoladas, com exceção de C. glabrata, C. lusitaniae e C. guilliermondii. Resistência ao fluconazol e anfotericina B foi observada em isolados de C. albicans, C. glabrata, C. parapsilosis, C. krusei, e K. ohmeri. CONCLUSÃO: Os pacientes HIV-positivos são colonizados por espécies únicas ou múltiplas de levedura que ocasionalmente são resistentes ao fluconazol ou anfotericina B.


Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Anfotericina B/farmacologia , Antifúngicos/farmacologia , Candida/classificação , Candidíase Bucal/microbiologia , Fluconazol/farmacologia , Soropositividade para HIV/microbiologia , Candida/efeitos dos fármacos , Candida/enzimologia , Testes de Sensibilidade Microbiana , Peptídeo Hidrolases/metabolismo , Fosfolipases/metabolismo
10.
Rev. Soc. Bras. Med. Trop ; 44(3): 334-338, May-June 2011. tab
Artigo em Inglês | LILACS | ID: lil-593358

RESUMO

INTRODUCTION: Candida yeasts are commensals; however, if the balance of normal flora is disrupted or the immune defenses are compromised, Candida species can cause disease manifestations. Several attributes contribute to the virulence and pathogenicity of Candida, including the production of extracellular hydrolytic enzymes, particularly phospholipase and proteinase. This study aimed to investigate the in vitro activity of phospholipases and acid proteinases in clinical isolates of Candida spp. METHODS: Eighty-two isolates from hospitalized patients collected from various sites of origin were analyzed. Phospholipase production was performed in egg yolk medium and the production of proteinase was verified in a medium containing bovine serum albumin. The study was performed in triplicate. RESULTS: Fifty-six (68.3 percent) of isolates tested were phospholipase positive and 16 (44.4 percent) were positive for proteinase activity. C. tropicalis was the species with the highest number of positive isolates for phospholipase (91.7 percent). Statistically significant differences were observed in relation to production of phospholipases among species (p<0,0001) and among the strains from different sites of origin (p=0.014). Regarding the production of acid protease, the isolates of C. parapsilosis tested presented a larger number of producers (69.2 percent). Among the species analyzed, the percentage of protease producing isolates did not differ statistically (χ2=1.9 p=0.5901 (χ2=1.9 p=0.5901). CONCLUSIONS: The majority of C. non-albicans and all C. albicans isolates were great producers of hydrolytic enzymes and, consequently, might be able to cause infection under favorable conditions.


INTRODUÇÃO: Candida são leveduras comensais, porém, se o equilíbrio da flora normal for interrompido ou as defesas imunitárias estiverem comprometidas, espécies de Candida podem causar manifestações de doença. Vários atributos contribuem na virulência e patogenicidade de Candida, inclusive a produção de enzimas extracelulares hidrolíticas, especialmente fosfolipases e proteinases. O objetivo deste estudo foi verificar a atividade in vitro de fosfolipases e proteinases ácidas em isolados clínicos de Candida spp. MÉTODOS: Oitenta e dois isolados provenientes de pacientes hospitalizados coletados a partir de sítios de origem diversos foram analisados. A produção de fosfolipase foi verificada em meio egg yolk e a de proteinase em meio contendo soro albumina bovina. O estudo foi feito em triplicata. RESULTADOS: Cinquenta e seis (68,3 por cento) dos isolados testados apresentaram atividade de fosfolipase positiva e 16 (44,4 por cento) foram positivos para atividade de proteinase. C. tropicalis foi a espécie que apresentou o maior número de isolados positivos para fosfolipases (91,7 por cento). Diferenças estatisticamente significantes em relação à produção de fosfolipases entre as espécies e entre as cepas provenientes de diferentes sítios de origem foram detectadas. Quanto à produção de proteinases ácidas, os isolados de C. parapsilosis testados foram os maiores produtores (69,2 por cento). Entre as espécies analisadas, a porcentagem de produção de proteinase entre os isolados não diferiu estatisticamente (χ2=1.9 p=0.5901 (χ2=1.9 p=0.5901). CONCLUSÕES: A maioria dos isolados de C. não-albicans, assim como os de C. albicans, foram grandes produtores de enzimas hidrolíticas e, consequentemente, podem ser capazes de causar infecção em condições adequadas.


Assuntos
Animais , Bovinos , Humanos , Candida/enzimologia , Peptídeo Hidrolases/biossíntese , Fosfolipases/biossíntese , Candida/classificação , Candida/isolamento & purificação
11.
Rev. Inst. Adolfo Lutz ; 70(2): 101-105, abr.-jun. 2011. tab, graf
Artigo em Inglês | LILACS, Sec. Est. Saúde SP, SESSP-CTDPROD, Sec. Est. Saúde SP, SESSP-ACVSES, SESSP-IALPROD, Sec. Est. Saúde SP, SESSP-IALACERVO | ID: lil-620818

RESUMO

Production of exoenzymes, specifically the proteinase and phospholipase, is considered one of the most important of pathogenicity mechanisms of C. albicans, which is crucial for tissue invasion. This study aimed at evaluating the production of these exoenzymes in 50 oral C. albicans isolates from HIV-positive (HIV+) patients treated with highly active anti-retroviral therapy (HAART), and from 50 control individuals. For testing the production of phospholipase and proteinase, the culture media containing egg yolk and bovine albumin were used, respectively. The results were obtained by measuring the diameter of the colony and divided by the diameter of colony plus the precipitation zone, defined as Pz. Data were statistically analyzed by Student’s t test (5). Statistically significant difference (p = 0.001) was observed between the mean values of Pz for proteinase in isolates from HIV+ patients (Pz = 0.358±0.295) and from control group (Pz = 0.660±0.370). The same results were observed for phospholipase production (Pz = 0.399±0.227 for HIV+ group; Pz =0.635±0.292 control group). Both enzymes were highly produced by C. albicans isolated from HIV+ patients when compared with those secreted by C. albicans obtained from control group, suggesting that HAART did not reduce the secretion of these enzymes by this pathogenic fungus infecting HIV+ patients.


Assuntos
Humanos , Masculino , Feminino , HIV , Terapia Antirretroviral de Alta Atividade , Boca , Candida albicans , Fosfolipases , Peptídeo Hidrolases
12.
Rev. patol. trop ; 39(2): 83-89, abr.-jun. 2010. tab
Artigo em Português | LILACS | ID: lil-561511

RESUMO

A criptococose é uma micose sistêmica com caráter oportunista, que pode acometer homens e diferentes espécies animais, principalmente em casos de imunodepressão. A levedura penetra pela via respiratória, pode disseminar-se por via hematógena e atingir o sistema nervoso central. Objetivando o isolamento da levedura Cryptococcus neoformans a partir de sistema nervoso central de cães na cidade do Rio de Janeiro, procederam-se coleta de amostras de cérebro no Laboratório de Diagnóstico de Raiva do Instituto Municipal de Medicina Veterinária Jorge Vaitsman, no bairro da Mangueira, Rio de Janeiro. As etapas de isolamento, identificação e avaliação da produção de protease e fosfolipase ocorreram no Laboratório de Leveduras Patogênicas e Ambientais da Universidade Federal Rural do Rio de Janeiro (LLPA-UFRRJ). Para a avaliação de protease, utilizou-se meio de cultura contendo albumina bovina e para avaliação de fosfolipase, o meio utilizado compunha-se de gema de ovo e CaCi2. Foram utilizadas 166 amostras de sistema nervoso central, com quatro isolamentos positivos para o gênero, sendo um de Cryptococcus laurentii, dois de Cryptococcus neoformans e um de Cryptococcus gattii, todos produtores de protease e fosfolipase.


Assuntos
Criptococose , Cryptococcus , Cryptococcus neoformans/isolamento & purificação , Peptídeo Hidrolases , Fosfolipases
13.
Rev. Inst. Med. Trop. Säo Paulo ; 52(3): 139-143, May-June 2010. tab
Artigo em Inglês | LILACS | ID: lil-550344

RESUMO

Phospholipase and proteinase production and the ability of adhesion to buccal epithelial cells (BEC) of 112 Candida isolates originated from oral cavity of HIV infected patients and from blood and catheter of intensive care unit patients were investigated. The proteinase production was detected by inoculation into bovine serum albumin (BSA) agar and the phospholipase activity was performed using egg yolk emulsion. A yeast suspension of each test strain was incubated with buccal epithelial cells and the number of adherence yeast to epithelial cells was counted. A percentage of 88.1 percent and 55.9 percent of Candida albicans and 69.8 percent and 37.7 percent of non-albicans Candida isolates produced proteinase and phospholipase, respectively. Non-albicans Candida isolated from catheter were more proteolytic than C. albicans isolates. Blood isolates were more proteolytic than catheter and oral cavity isolates while oral cavity isolates produced more phospholipase than those from blood and catheter. C. albicans isolates from oral cavity and from catheter were more adherent to BEC than non-albicans Candida isolates, but the adhesion was not different among the three sources analyzed. The results indicated differences in the production of phospholipase and proteinase and in the ability of adhesion to BEC among Candida spp. isolates from different sources. This study suggests that the pathogenicity of Candida can be correlated with the infected site.


A produção de proteinase e fosfolipase e habilidade de adesão à célula epitelial bucal de 112 isolados de Candida originadas da cavidade bucal de pacientes infectados pelo HIV e de sangue e cateter de pacientes hospitalizados foram investigados. A produção de proteinase foi detectada por inoculação em ágar soro albumina bovina e a atividade de fosfolipase foi realizada usando emulsão de gema de ovo. A suspensão de levedura de cada isolado foi incubada com célula epitelial e o número de leveduras aderidas a célula epitelial foi contada. Uma porcentagem de 88,1 e 55,9 por cento de C. albicans e 69,8 e 37,7 por cento de isolados de Candida não albicans produziram proteinase e fosfolipase, respectivamente. Candida não albicans obtidas do cateter foram mais proteolíticos que isolados de Candida albicans (p < 0,001). Isolados do sangue foram mais proteolíticos do que isolados do cateter e cavidade bucal, enquanto isolados da cavidade bucal produziram mais fosfolipase do que aqueles isolados do sangue e cateter. C. albicans isoladas da cavidade bucal e do cateter foram mais aderentes à célula epitelial bucal do que isolados de Candida não albicans, mas não houve diferença na adesão entre os três locais analisados. Os resultados indicaram diferenças na produção de fosfolipase e proteinase e na habilidade de adesão à célula epitelial bucal entre os isolados de Candida das diferentes fontes. Este estudo sugere que a patogenicidade de Candida spp pode estar correlacionada ao local infectado.


Assuntos
Humanos , Ácido Aspártico Proteases/biossíntese , Aderência Bacteriana/fisiologia , Candida/enzimologia , Candida/fisiologia , Fosfolipases/biossíntese , Candida/isolamento & purificação , Cateteres de Demora/microbiologia , Células Epiteliais/microbiologia , Infecções por HIV/microbiologia , Boca/microbiologia
14.
Rev. Soc. Bras. Med. Trop ; 43(2): 205-206, Mar.-Apr. 2010.
Artigo em Inglês | LILACS | ID: lil-545778

RESUMO

INTRODUCTION: The phospholipase activity in Candida albicans and Candida dubliniensis isolated from oral candidiasis cases were studied. METHODS: The phospholipase activity was evaluated in egg yolk agar. RESULTS: All the C. albicans isolates (n = 48) showed phospholipase activity (mean Pz = 0.66). However, none of the C. dubliniensis isolates (n = 24) showed this activity. CONCLUSIONS: The authors discuss whether these findings are a true characteristic of C. dubliniensis or a consequence of the methodology employed, which includes the possibility that NaCl may have inhibited the enzymatic activity of C. dubliniensis.


INTRODUÇÃO: Avaliou-se a atividade fosfolipásica em Candida albicans e Candida dubliniensis isoladas de casos de candidíase oral. MÉTODOS: A atividade de fosfolipase foi avaliada em ágar gema de ovo. RESULTADOS: Todos os isolados de C. albicans (nº = 48) evidenciaram atividade fosfolipásica (média Pz = 0.66). Todavia, nenhum isolado de C. dubliniensis (nº= 24) demonstrou esta atividade. CONCLUSÕES: Os autores discutem se estes achados são uma característica verdadeira de C. dubliniensis ou uma conseqüência da metodologia empregada, a qual inclui a possibilidade de que o NaCl seja inibidor da atividade enzimática de C. dubliniensis.


Assuntos
Humanos , Candida/enzimologia , Candidíase Bucal/microbiologia , Fosfolipases/metabolismo , Candida albicans/enzimologia , Candida albicans/patogenicidade , Candida/patogenicidade , Fosfolipases/análise
15.
Braz. j. microbiol ; 41(1): 19-23, Jan.-Mar. 2010. tab
Artigo em Inglês | LILACS | ID: lil-531728

RESUMO

Patients suffering of diseases that affect central nervous system may be considered more susceptible to the infectious diseases of mouth. Sixty-nine patients suffering of cerebral palsy, Down's syndrome and metal retardation were submitted to saliva examination for the presence of Candida spp. before and after a procedure of dental cleaning. The isolates were submitted to assay for verifying phospholipase production. 55.10 percent of the patients provided isolation of Candida spp. The frequency of isolation obtained before dental procedure was: C. albicans (83.33 percent), C. krusei (8.33 percent) and C. kefyr, C. parapsilosis and C. glabrata (2.78 percent each). The frequency after the procedure was: C. albicans (68.57 percent), C. parapsilosis (11.43 percent), C. krusei and C. kefyr (8.57 percent each) and Candida glabrata (2.86 percent). We verified significantly difference (p < 0.01) between populations obtained at the two examinations. Phospholipase production was verified only among C. albicans strains and the proportion of producers was higher when testing isolates obtained after dental cleaning procedure. Studies focused on Candida spp. isolation are useful for better comprehension of the role of these yeasts on the oral flora from patients with cerebral palsy, Down's syndrome and metal retardation.


Assuntos
Humanos , Criança , Candidíase Bucal , Paralisia Cerebral , Candida/isolamento & purificação , Síndrome de Down , Fosfolipases/análise , Fosfolipases/isolamento & purificação , Deficiência Intelectual , Escovação Dentária , Técnicas e Procedimentos Diagnósticos , Epidemiologia , Métodos
16.
Rev. Soc. Bras. Med. Trop ; 42(1): 63-66, Jan.-Feb. 2009. tab
Artigo em Português | LILACS | ID: lil-507367

RESUMO

Enzimas hidrolíticas secretadas por fungos têm um papel importante na patogenicidade das infecções. Objetivando avaliar a atividade enzimática foram testados 31 isolados de Acremonium mantidos na Coleção de Culturas University Recife Mycology. Fragmentos das culturas foram transferidos para caldo glicosado para reativação e posterior crescimento em meio ágar batata dextrose, para verificar viabilidade, pureza e confirmação taxonômica pela observação das características macroscópicas e microscópicas. Para detecção enzimática foram utilizados substratos de caseína do leite e gelatina para protease, amido para amilase e lecitina de soja para fosfolipase. Das 31 culturas, 26 (83,9 por cento) mantiveram-se viáveis e 24 (92,3 por cento) foram confirmadas taxonomicamente. Das 24 culturas, 12 (50 por cento) apresentaram atividade proteásica, duas (16,7 por cento) em caseína do leite, uma (8,3 por cento) em gelatina e nove (75 por cento) em ambos os substratos; 16 (66,7 por cento) degradaram amido. Nenhuma cultura apresentou atividade fosfolipásica. Conclui-se que espécies de Acremonium são capazes de produzir enzimas envolvidas na patogenicidade das infecções fúngicas.


Hydrolytic enzymes secreted by fungi play an important role in the pathogenesis of infection. With the aim of evaluating the enzymatic activity, 31 isolates of Acremonium stored in the University of Recife Mycology (URM) Culture Collection were tested. Culture fragments were transferred to glycoside broth for reactivation and further growth in potato dextrose agar medium in order to investigate viability and purity and to confirm the taxonomy through observing the macroscopic and microscopic characteristics. To detect enzymes, milk casein and gelatin were used as substrates for proteinase, starch for amylase and soy lecithin for phospholipase. Among the 31 cultures, 26 (83.9 percent) remained viable and 24 (92.3 percent) were confirmed taxonomically. Out of these 24 cultures, 12 (50 percent) presented proteinase activity, of which two (16.7 percent) were on milk casein, one (8.3 percent) on gelatin and nine (75 percent) on both substrates; 16 (66.7 percent) degraded starch. None of the cultures presented phospholipase activity. It was concluded that Acremonium species are able to produce enzymes that are involved in the pathogenicity of fungal infections.


Assuntos
Acremonium/enzimologia , Amilases/biossíntese , Peptídeo Hidrolases/biossíntese , Fosfolipases/biossíntese , Acremonium/classificação , Acremonium/crescimento & desenvolvimento , Óleo Mineral , Preservação Biológica/métodos
17.
Bol. micol ; 23: 21-25, dic. 2008. tab
Artigo em Espanhol | LILACS | ID: lil-585729

RESUMO

Se detectó la actividad de fosfolipasa en 19 cepas clínicas y 17 aviarias de C. neoformans var. neoformans, usando Agar Sabouraud con yema de huevo, incubándose a 37ºC por 5 días. Se determinó el índice Pz estableciéndose los siguientes rangos: Pz muy alto (0.9-1), alto (0.89-0.80), bajo (0.79-0.70) y muy bajo (<0.69). El 84 por ciento de las cepas clínicas presentaron índice Pz muy bajo, el 5 por ciento bajo y 11 por ciento muy alto. Mientras en las cepas aviarias el 82 por ciento presentaron índice muy bajo y un 18 por ciento muy alto. Los valores de Pz promedio fueron muy bajos en todos los aislamientos, sin existir diferencias significativas (p > 0,05) entre las cepas clínicas y aviarias, lo que implica una alta actividad enzimática. La susceptibilidad in vitro a Fluconazol se realizó por el método de difusión con discos y el 89,5 por ciento de las cepas clínicas fueron sensibles y el 10,5 por ciento resistentes, mientras en las cepas aviarias, el 59 por ciento fueron sensibles, 29 por ciento sensible dosis dependiente y un12 por ciento resistentes.


Phospholipase activity was determined to 19 clinical and 17 aviars trains of C. neoformans var. neoformans, incubating the yeast for 5 days at 37º C on Sabouraud Agar supplemented with egg yolk. Pz values were determined and the following ranges were established: very high (0.9-1), high (0.89-0.80), low (0.79-0.70) and very low (<0.69). The 84 percent of the clinical isolates showed Pz values very low (5 percent) and 11 percent very high. On the other hand, the 82 percent of the aviars strains presented Pz values very low and 18 percent very high. Average Pz values were very low in all isolates , there were no statically significant differences (p>0.05) implying a high enzymatic activity. Susceptibility in vitro testing to Fluconazole was performed by a disk diffusion method (M44-A).The 89.5 percent of the clinical isolates were susceptible and 10.5 percent resistant, while in avian strains, 59 por ciento were susceptible, 29 percent susceptible dose dependent and 12 percent resistant.


Assuntos
Cryptococcus neoformans/isolamento & purificação , Cryptococcus neoformans/crescimento & desenvolvimento , Cryptococcus neoformans/enzimologia , Fluconazol , Testes de Sensibilidade Microbiana , Fosfolipases
18.
Rev. bras. anal. clin ; 40(3): 205-207, 2008. tab
Artigo em Português | LILACS | ID: lil-541905

RESUMO

A candidíase oral é uma doença oportunista que acomete principalmente indivíduos imunocomprometidos. Entre os fatores responsáveis pelo aumento da prevalência de infecções por Candida spp., está a quimioterapia antineoplásica, que favorece a instalação de patógenos oportunistas, bem como a proliferação da microbiota, por causar um estado de comprometimento do sistemaimune. Este trabalho teve como objetivos avaliar a freqüência de Candida spp. em isolados da mucosa oral de pacientes submetidosà quimioterapia, além da identificação das espécies e atividade de fosfolipase. As amostras de swabs orais de 25 pacientes foram semeadas em ágar de Sabouraud dextrose, incubadas por 48 horas, a 37 °C, para posterior identificação por testes micológicos. Dez pacientes apresentaram leveduras do gênero Candida sendo todas C. albicans produtoras de fosfolipase. Possivelmente, os efeitos daquimioterapia estão relacionados com o aparecimento de infecções fúngicas nos pacientes analisados.


Oral candidiasis is an oportunist disease that catches overall immunecompromised individuals. Among the factors responsible for increase of Candida infection prevalence, the anticancer chemotherapy leads to oportunistic infections, as well as the proliferation of normal microbiota. This treatment cause a immune system compromisement. The aims of this paper was evaluate the Candida spp. frequency in oral mucus isolated of chemotherapy treat patients, beyond the identification and phospholipase enzimatic activity. The oral swabs samples of 25 patients were inoculated in Sabouraud agar dextrose, maintained for 48 hours at 37°C to mycologicaltests. Ten patients had conferred leavenings of Candida spp., being all C. albicans phospholipase producers. Possibly, the chemotherapy effects are related with the apearance of fungal infections in the analyzed patients.


Assuntos
Humanos , Candidíase Bucal , Candida/isolamento & purificação , Tratamento Farmacológico , Mucosa Bucal , Fosfolipases
19.
Rev. Inst. Med. Trop. Säo Paulo ; 49(6): 349-353, Nov.-Dec. 2007. tab
Artigo em Inglês | LILACS | ID: lil-470516

RESUMO

The study involved 100 yeast isolates, obtained from urine samples provided by a Public Pediatric Hospital of São Paulo, Brazil, from 1999 to 2004. The most frequent species was Candida albicans, followed by C. tropicalis, C. glabrata and C. parapsilosis. In regard to virulence, 97 percent of the isolates showed index 3 for proteinase and 63 percent index 2 for phospholipase. The most frequent killer biotypes were 511 and 888.


Estudou-se 100 amostras de leveduras, isoladas de urina, provenientes de Hospital Público Infantil de São Paulo Brasil, no período de 1999-2004. A espécie mais freqüente foi Candida albicans, seguida de C. tropicalis, C. glabrata e C. parapsilosis. Em relação à virulência, 97 por cento dos isolados apresentaram índice 3 para proteinase e, 63 por cento índice 2 para fosfolipase. Os biótipos "killer" mais freqüentes foram o 511 e 888.


Assuntos
Criança , Pré-Escolar , Humanos , Lactente , Recém-Nascido , Candida/classificação , Candidíase/urina , Infecções Urinárias/microbiologia , Brasil , Candida/enzimologia , Candida/isolamento & purificação , Candidíase/microbiologia , Peptídeo Hidrolases/metabolismo , Fosfolipases/metabolismo
20.
Rev. Soc. Bras. Med. Trop ; 40(3): 354-355, maio-jun. 2007. tab
Artigo em Português | LILACS | ID: lil-456337

RESUMO

A habilidade de Candida spp secretar enzimas extracelulares e slime tem sido associada como fatores de patogenicidade. Do total de 37 cepas de Candida sp, 100 por cento foram produtoras de proteinase, 83,8 por cento fosfolipase, 64,9 por cento slime e 100 por cento sensíveis ao fluconazol e itraconazol. Foram encontradas 17 tipagens (enzima/slime). Esta metodologia apresentou um bom índice discriminatório (D=0,93) podendo ser utilizado na caracterização fenotípica das leveduras.


Abilith of Candida spp to secrete extracellular enzymes and slime has been associated as pathogenicity factors. Out of a total of 37 strains of Candida sp, 100 percent were proteinase producers, 83.8 percent were phospholipase producers, 64.9 percent were slime producers and 100 percent were sensitive to fluconazole and itraconazole. Seventeen typings (enzymes/slime) were found. This methodology presented a good discrimination rate (D = 0.93) and could be used for phenotypic characterization of yeasts.


Assuntos
Humanos , Antifúngicos/farmacologia , Biofilmes/crescimento & desenvolvimento , Candida/efeitos dos fármacos , Peptídeo Hidrolases/metabolismo , Fosfolipases/metabolismo , Biofilmes/efeitos dos fármacos , Candida/enzimologia , Fluconazol/farmacologia , Itraconazol/farmacologia , Testes de Sensibilidade Microbiana , Peptídeo Hidrolases/efeitos dos fármacos , Fosfolipases/efeitos dos fármacos
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