Natalina Takahashi de Melo

Comprometimento da medula óssea e eosinofilia na paracoccidioidomicose

Sao descritos 3 casos de paracoccidioidomicose com a forma aguda da doenca, nos quais formas leveduriformes de Paracoccidioides brasiliensis foram visualizadas ao exame direto de medula ossea, sendo a cultura tambem positiva em um caso. Salienta-se o acometimento do sistema fagocitico-mononuclear e a ausencia de resposta as provas cutâneas de hipersensibilidade tardia a antigenos microbianos e de P. brasiliensis em todos, bem como a gravidade do quadro clinico e lesoes osseas generalizadas em um caso, com 20.260 eosinofilos/mm3 no sangue periferico. Os autores discutem o possivel papel do eosinofilo na interacao hospedeiro-parasita na paracoccidioidomicose, sugerindo que a ativacao de subpopulacao TH 2 e o aumento de secrecao de IL 5 e de GM-CSF possam estar relacionados a grande eosinofilia presente no caso mais grave

Eumicetoma de grãos pretos por Madurella grisea: registro de dois casos

Two cases of black grains eumycotic mycetoma, occurring on a foot, are reported. Both proceeded from the State of Bahia (Brazil), and in both the etiologic agent was Madurella grisea Mackinnon et al., 1949. The grains structure as well as the micromorphologic characteristics of the fungus in saprophytic life were studied. It is the Authors belief that these observations correspond to the 7th and 8th cases reported in the Brazilian medical literature. The Authors do consider the following Madurella species as nomen dubium or nomina confusa: M. ramiroi, M. oswaldoi, M. bovoi, M. tozeuri, M. mansonii, M. brumpti, M. reynieri, M. americana, M. lackawanna e M. ikedae and the same for Rubromadurella mycetomi. The only valid species must be Madurella mycetomatis McGinnis, 1980 (=Madurella mycetomi Brumpt, 1905) and Madurella grisea Mackinnon et al., 1949. Treatment with itraconazole in both reported cases, for a 3 month duration, did not produce any regression of the lesions, the clinical improvement being meager.Os Autores registram dois casos de eumicetoma de graos pretos, com localizacao podal, procedentes da Bahia, provocados por Madurella grisea Mackinnon et al., 1949. Sao estudadas a estrutura dos graos, bem como as caracteristicas micromorfologicas do fungo em vida saprofitica. Acreditam os Autores que estas observacoes correspondem ao setimo e oitavo casos registrados na literatura do pais, provocadas por este fungo. Os Autores consideram nomen dubium ou nomina confusa as seguintes especies de Madurella: M. ramiroi, M. oswaldoi, M. bovoi, M. tozeuri, M. mansonii, M. brumpti, M. reynieri, M. americana, M. lackawanna e M. ikedae, o mesmo ocorrendo com a chamada Rubromadurella mycetomi. As unicas especies validas sao Madurella mycetomatis McGinnis, 1980 (=Madurella mycetomi Brumpt, 1905) e Madurella grisea Mackinnon et al., 1949. Nos dois casos registrados o tratamento com itraconazol, por um periodo de 3 meses nao fez regredir as lesoes, havendo ligeira melhora clinica.

Molecular study of archival fungal strains isolated from cases of lacaziosis (Jorge Lobo's disease).

Lacazia loboi, the aetiological agent of lacaziosis (Jorge Lobos disease), is an uncultivated anomalous fungal microbe closely related to Paracoccidioides brasiliensis, both restricted Latin American Pathogens. Early reports suggesting that L. loboi had been isolated in pure culture from cases of lacaziosis, only added more confusion to the already confusing aetiology of this disease. These strains were later identified as unusual contaminants and some of them as P. brasiliensis. Recent phylogenetic analysis grouped L. loboi as the sister taxon to P. brasiliensis, thus it was postulated that the original P. brasiliensis strains recovered from cases of lacaziosis, could be the aetiological agent of the disease. Using molecular methodologies, we investigated the archival P. brasiliensis isolate No. 525 from a case of lacaziosis, as well as other archival isolates, identified earlier as common contaminants, all recovered from similar cases of the disease. Phylogenetic analysis, using the 18S small subunit rDNA sequences of these isolates showed that strain No. 525 was a typical P. brasiliensis isolate and the other studied strains were indeed contaminants. This study unequivocally indicates that the aetiological agent of lacaziosis is yet to be cultured.

Dermatophytosis caused by Trichophyton raubitschekii. Report of the first case in São Paulo, Brazil

The authors report the first case of dermatophytosis caused by Trichophyton raubitschekii in a patient from the State of São Paulo with Tinea corporis lesions localized on the buttocks. Culture on Sabouraud-agar with cycloheximide permitted the isolation and identification of the fungus, and the diagnosis was confirmed by Dr. Lynne Sigler, University of Alberta, Canada. Systemic treatment with fluconazole, 150 mg/week for 4 weeks, in combination with topical treatment with isoconazole initially yielded favorable results, with recurrence of the lesions after the medication was discontinued. This is the fifth case of this dermatophytosis published in the Brazilian medical literature.

Paracoccidioides brasiliensis: a mycologic and immunochemical study of a sample isolated from an armadillo (Dasipus novencinctus)

A sample of P. brasiliensis isolated from the spleen and the liver of an armadillo (Dasipus novencinctus) has been analysed under a mycological and immunochemical viewpoint. The armadillo was captured in an area of Tucuruí (State of Pará, Brazil), the animal being already established as an enzootic reservoir of P. brasiliensis at that region of the country. This sample maintained in the fungal collection of the Tropical Medicine Institute of São Paulo (Brazil) numbered 135, has got all the characteristics of P. brasiliensis, with a strong antigenic power and low virulence for guinea-pigs and Wistar rats. The specific exoantigen of P. brasiliensis--the glycoprotein with a molecular weight of 43 kDa--was easily demonstrated with double immunodiffusion, immunoelectrophoresis, SDS-PAGE and immunobloting techniques.

Reactivity of anti-gp43 antibodies from Paracoccidioides brasiliensis antiserum with extracts from cutaneous lesions of Lobo's disease. Preliminary note.

We demonstrated through several immunochemical tests the presence of gp43 from P. brasiliensis in extracts of cutaneous lesions from Jorge Lobos disease. This glicoprotein is one of the immunodominant antigens in this species, and is used to identify it. The demonstration of gp43 tissues infected by the agent of Jorge Lobos disease is an additional evidence for classifying it in the genera Paracoccidioides, species loboi.

In vitro sensitivity of dermatophytes to urea

OBJECTIVE Urea is commonly used as a keratolytic substance in the treatment of onychomycoses to improve the penetration of antifungal drugs in the lesion sites. The aim of the present study was to investigate the inhibitory action of urea on samples of dermatophytes in vitro. METHOD Minimum inhibitory concentration of urea was determined for 31 samples of dermatophytes cultured in Sabouraud-dextrose broth containing different concentrations (7.5% up to 40%) of urea. Absence of growth was the criterion adopted to determine the minimum inhibitory concentration. RESULTS The majority of samples (87%) were sensitive to urea at 12.5%, or less. 2 isolates of Trichophyton tonsurans and 2 of Trichophyton rubrum required 30%, and 40% urea, respectively, to be completely inhibited. CONCLUSION In vitro results demonstrate inhibitory activity of urea on dermatophytes, suggesting that it could be used as an adjuvant in topical treatments.

Neurocriptococose por Cryptococcus neoformans não capsulado

Os autores registram um caso de neurocriptococose em paciente HIV-negativo, por Cryptococcus neoformans acapsulado ou deficiente em capsula. O quadro neurologico era de meningoencefalite subaguda, compativel ao diagnostico de neurotuberculose, pelo exame do liquido cefalorraqueano (LCR), Estruturas leveduriformes foram encontradas no interior de macrofagos, ao exame citomorfologico do LCR. Cultivo do sedimento do LCR revelou a presenca de Cryptococcus neoformans nao capsulado (identificacao bioquimica). A inoculacao da amostra em camundongo, por via intraperitoneal, permitiu a producao de capsula que desaparecia em cultivos. Foi estudada a micromorfologia do fungo a microscopia eletronica de varredura. A evolucao foi favoravel com o emprego da anfotericina B associada a 5-fluorocitosina. Nao foi caracterizada a variadade de Criptococcus neoformans agente do processo.The case of a patient with meningoencephafalitis due to a nonencapsulated strain of Cryptococcus neoformans is reported; he had no risk factors for the disease or AIDS. Clinical examination showed a chronic meningoencephalitis first diagnosed as tuberculosis. In the second cerebrospinal fluid (CSF) examination after a week from admission yeasts appeared inside macrophage cells. CSF culture in Sabouraud medium disclosed nonencapsulated Cryptococcus neoformans (biochemical identification). Sample inoculation in mouse (intraperitoneal) evidenced a capsule that disappeared in several consecutive cultures. The morphology of the yeast was studied by electronic microscopy. After treatment with amphotericin B and 5-fluorocytosine the patient had a favorable evolution. The significance of capsular material is discussed.

Paracoccidioides brasiliensis: A MYCOLOGIC AND IMMUNOCHEMICAL STUDY OF TWO STRAINS

Os Autores estudaram do ponto de vista micologico, imunoquimico e de sua biologia molecular, duas amostras de Paracoccidioides brasiliensis, uma isolada do solo, no municipio de IBIA (MG) por Silva-Vergara et al. (l996,1998)20,21 denominada IBIA e outra, BAT, cultivada de um caso humano de paracoccidioidomicose em Ribeirao Preto (SP) por Freitas da Silva (l996)6. Tais amostras apresentam colonias cotonosa (M) e leveduriforme (L ou Y), sendo patogenicas para cobaios inoculados por via testicular, produzindo orquite granulomatosa e/ou supurativa. Do ponto de vista imunoquimico, atraves de provas de Imunodifusao dupla, Imunoeletroforese e Western Blotting, foi demonstrada a presenca da gp43. A sequencia de nucleotideos do DNA de tais amostras, atraves do sequenciamento de 761 bases, revelou homologia de 100% com amostra padrao de P. brasiliensis, o mesmo ocorrendo com tres amostras humanas, uma isolada de fezes de pinguim e outra de racao alimentar para caes contaminada com terra. Sao discutidos varios aspectos dos resultados obtidos, comparando-os com alguns dados da literatura.The authors conducted a mycologic, immunochemical and molecular biology study on two strains of Paracoccidioides brasiliensis, one of them, called IBIA, isolated from soil in the municipality of IBIA (Minas Gerais) by Silva-Vergara et al. (1996, 1998), and the other, BAT, cultivated from a human case of paracoccidioidomycosis in Ribeirão Preto (São Paulo/Brazil) by Freitas Da Silva (1996). Both strains showed cotton-like (M) and yeast-like (Y) forms and were pathogenic for testicularly inoculated guinea pigs, producing granulomatous and/or suppurative orchitis. Immunochemically was demonstrated the presence of gp43 by double immunodiffusion, immunoelectrophoresis and immunoblotting.

In vitro action of some disinfectants on Paracoccidioides brasiliensis yeast forms

The fungicidal action of sodium hypochlorite (0.3, 1, 2.5, 5 and 10%); formaldehyde (2, 5, and 10%); and ethyl alcohol (70%) on yeast forms of Paracoccidioides brasiliensis Pb 18 and a newly-isolated Goiana strain was described. Contact between the fungus and the disinfectants was maintained for 1, 2, 24, 48 and 72 hours at room temperature. Viability was evaluated by the fluorescein diacetate-ethidium bromide treatment, culture in solid and liquid media (36 degrees C and 26 degrees C); yeast to mycelial germination at room temperature; and radiometric study of metabolic activity. All concentrations of disinfectants were found to be effective in inactivating Pb 18 and Goiana strains, except for the 1-hour contact with 2% formaldehyde, in which fluorescein diacetate-ethidium bromide treatment was found to reveal 40 and 27% of viable cells, respectively. The yeast to mycelial germination method was considered to reveal faster and similar results as compared to culture in solid and liquid media.