Estudos de peptidases secretadas por Phialophora verrucosa: efeito de inibidores proteolíticos/agentes quelantes na biologia celular / Studies peptidases secreted by Phialophora verrucosa: effects of proteolytic inhibitors / chelating agents in cell biology

Phialophora verrucosa é um dos agentes etiológicos da cromoblastomicose, uma micose que afeta os tecidos cutâneos e subcutâneos. Esta doença é crônica, recidivante e de difícil tratamento. Vários estudos têm demonstrado que peptidases secretadas estão associadas a importantes processos biológicos. Neste trabalho, foi relatada a presença de atividade peptidásica extracelular em P. verrucosa. A peptidase secretada por este fungo apresentou ótima atividade em condições acídicas e à 37ºC. A atividade proteolítica foi completamente inibida por 1,10-Fenantrolina (1,10-Fen) e EGTA, sugerindo a presença de metalopeptidase em P. verrucosa. Zinco (Zn2+) foi o único metal capaz de estimular a atividade peptidásica, de forma dose dependente. Esta atividade enzimática gerou a hidrólise de proteínas do soro e de matriz extracelular como soro albumina humana (HSA), imunoglobulina G (IgG), fibrinogênio, colágeno (tipo I e IV), fibronectina e laminina. Queratina também foi eficientemente hidrolisada pela atividade peptidásica de P. verrucosa. Esses dados sugerem o envolvimento dessa enzima na disseminação fúngica e evasão do sistema imune do hospedeiro. O crescimento de P. verrucosa na presença de 1,10-Fen e seus derivados complexados a metais, como [Cu(fendio)3](ClO4)2•4H2O e [Ag(fendio)2]ClO4 (Fen-Cu2+ e Fen-Ag+, respectivamente) e a forma livre de metal (Fendio) foi também testado. 1,10-Fen foi o composto mais efetivo na inibição da proliferação fúngica. Fen-Cu2+, Fen-Ag+ e Fendio também apresentaram intensa atividade antimicrobiano. O complexo Cu2+ e Fendio foram fungicidas, enquanto Fen-Ag+ demonstrou atividade fungistática. Conídios tratados com 1,10-Fen e seus derivados apresentaram importantes alterações morfológicas, indicativas de morte celular, como determinado por microscopia eletrônica de transmissão. Além disto, Fen-Cu2+ e Fen-Ag+ foram capazes de inibir a atividade proteolítica, enquanto Fendio não afetou a hidrólise do substrato proteico. 1,10-Fen e Fen-Cu2+ inibiram a filamentação de P. verrucosa. No entanto, Fen-Ag+ foi capaz de inibir a produção de esterol por este fungo. Os resultados também sugerem um efeito antifúngico sinérgico entre Fen-Cu2+ e anfotericina B (AMB). Nossos dados suportam a hipótese de que inibidores de metalopeptidases/agentes quelantes apresentam potencial para serem utilizados em modelos futuros para o tratamento da cromoblastomicose.

Chromoblastomycosis in Korea / 대한의진균학회지

The dematiaceous fungi, characterized by having melanin in their cell walls, have become significant due to an increasing cause of human skin disease in Korea. Chromoblastomycosis is subcutaneous mycotic disease that contains sclerotic cells or muriform cells and histologically show pseudoepitheliomatous hyperplasia with microabscess, and phaeohyphomycosis is a group of mycotic infections that contain dematiaceous yeast-like cells, pseudohypae-like elements, hyphae or combination of these form in tissue. Ten Korean cases of subcutaneous chromoblastomycosis have been reported to date, four males and five females, ranging in age from 37~68 (mean 53.8 years). Causative organisms of subcutaneous chromoblastomycosis were eight of Fonsecaea pedrosoi and one of Phialophora verrucosa. Nine cases of erythematous or verrucous plaque and one case of ulcer were present skin lesion, and most of exposed areas of the body except one case. Patients were treated with itraconazole, terbinafine, 5-flucytocine or amphotericin B.

Chromoblastomycosis Caused by Phialophora richardsiae

Chromoblastomycosis is a chronic fungal disease of the skin and subcutaneous tissues caused by a group of dematiaceous (black) fungi. The most common etiologic agents are Fonsecaea pedrosoi and Cladophialophora carrionii, both of which can be isolated from plant debris. The infection usually follows traumatic inoculation by a penetrating thorn or splinter wound. Several months after the injury, painless papules or nodules appear on the affected area; these papules then progress to scaly and verrucose plaques. We report a case of chromoblastomycosis caused by Phialophora richardsiae, which has been rarely associated with chromoblastomycosis. The case involved a 43-year-old male, who for the past 2 months had noted an erythematous, pustulous plaque that was somewhat dark brown in color on his right shin; the plaque also had intermittent purulent discharge and crust formation. On histopathological examination, chronic granulomatous inflammation and sclerotic cells were seen. The tissue fungus culture grew out the typical black fungi of P. richardsiae, which was confirmed by polymerase chain reaction. The patient has been treated with a combination of terbinafine and itraconazole for 3 months with a good clinical response.

Chromoblastomycosis: two cases of a disfiguring deep mycosis

Chromoblastomycosis is a chronic cutaneous and subcutaneous mycotic infection caused mostly by traumatic implantation of pigmented saprophytic moulds ubiquitous in the environment. If not diagnosed earlier, it has a chronic evolutional course that may cause several problems. We present two cases [father and a son] who were simultaneously infected with this uncommon fungal infection. Both had extensive face involvement. Diagnosis was considerably delayed and they were subjected to a myriad of empirical treatments

Chromoblastomycosis Caused by Rhinocladiella aquaspersa / 대한의진균학회지

Chromoblastomycosis is a rare disease in Korea and a total of 7 cases have been reported until now. The causative agents were Fonsecaea pedrosi in 5 cases and Phialophora verrucosa in 1 case. Rhinocladiella (R.) aquaspersa is a very rare etiologic agent and only a few cases of chromoblastomycosis due to this fungus have been reported in the Central and South Americas. We report a case of chromoblastomycosis caused by R. aquaspersa in a 52-year-old woman. The lesion was an erythematous, verrucous plaque on the abdomen, 2x1.3 cm in size, and had developed for 6 months before presentation without a traumatic history. She was successfully treated with itraconazole 200 mg/day for 4 months together with surgical excision without recurrence.

Ribosomal DNA Gene Analysis of Dematiaceous Fungi Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism / 대한피부과학회지

BACKGROUND: There are three kinds of diseases caused by dematiaceous fungi: chromoblastomycosis, phaeohyphomycosis, and eumycotic mycetoma. The dematiaceous fungi have been identified and classified by morphological, biochemical and physiological tests. Recently molecular analysis has been introduced to the field of medical mycology. OBJECTIVE: Ribosomal DNA gene analysis of dematiaceous fungi using polymerase chain reaction(PCR)-restriction fragment length polymorphism(RFLP) is investigated. METHODS: The dematiaceous fungal strains studied were eight clinical isolates of chromoblastomycosis and phaeohyphomycosis agents(3 strains of Fonsecaea pedrosoi, 2 strains of Exophiala dermatitidis, 1 strain of Exophiala jeanselmei, 1 strain of Phialophora verrucosa, 1 strain of Rhinocladiella aquaspersa) and 4 standard strains(F. pedrosoi IFM 4889, E. dermatitidis IFM 4828, P. verrucosa IFM 4928, R. aquaspersa IFM 4930). Total twelve strains of dematiaceous fungi were cultured on Sabouraud dextrose broth and their DNA was extracted by bead-beating method. PCR-RFLP of ribosomal gene small subunit(SSU rDNA) and internal transcribed spacer(ITS)1-ITS2 ribosomal regions using the primer pairs NS1-NS2 and ITS1-ITS4, respectively were done. RESULTS: The PCR product obtained from the SSU rDNA amplification was of approximately 603 bp and restriction profile was clustered into two genetically heterogenous groups, the first one formed by F. pedrosoi and the second one formed by other dematiaceous fungi. In contrast, the amplified PCR products of ITS regions ranged in sized from 580 to 620 bp and restriction profile was clustered into five genetically heterogenous groups and it can be possible to differentiate dematiaceous fungi. But one clinical isolate of F. pedrosoi showed intra-specific variability. CONCLUSION: The PCR-RFLP analysis of SSU rDNA and ITS regions provided useful information for identification and classification of dematiaceous fungi.

Genetic Diversity of Dematiaceous Fungi Using Random Amplified Polymorphic DNA / 대한의진균학회지

BACKGROUND: There are three kinds of diseases caused by dematiaceous fungi: chromoblastomycosis, phaeohyphomycosis, and eumycotic mycetoma. The dematiaceous fungi have been identified and classified by morphological, biochemical and physiological tests. Recently molecular analysis has been introduced to the field of medical mycology. OBJECTIVE: We investigated the genetic diversity of dematiaceous fungi using random amplified polymorphic DNA (RAPD). METHODS: The dematiaceous fungal strains studied were eight clinical isolates of chromoblastomycosis and phaeohyphomycosis agents (3 strains of Fonsecaea pedrosoi, 2 strains of Exophiala dermatitidis, 1 strain of Exophiala jeanselmei, 1 strain of Phialophora verrucosa, 1 strain of Rhinocladiella aquaspersa) and 4 standard strains (F. pedrosoi IFM 4889, E. dermatitidis IFM 4828, P. verrucosa IFM 4928, R. aquaspersa IFM 4930). Total twelve strains of dematiaceous fungi were cultured on Sabouraud's dextrose broth and their DNA were extracted by bead-beating method. RESULTS: The optimal condition for PCR was template DNA 0.025 mg and annealing temperature 39 degrees C. The RAPD analysis using OPA 10 primer (5'-GTGATCGCAG-3') of Operon kit showed different patterns among dematiaceous fungi. But one clinical isolate of F. pedrosoi showed intra-specific variability. CONCLUSION: The RAPD analysis is considered a rapid and reliable method for identification and classification of dematiaceous fungi if the procedure is carefully standardized with adequate primer.

Mycetoma and phaeohyphomycosis caused by Phialophora parasitica in Thailand.

Phaeohyphomycosis caused by Phialophora parasitica is rare and it has never been documented in Thailand. The first two Thai cases of phaeohyphomycosis caused by P. parasitica were recognized in early 1990 at Ramathibodi Hospital, Bangkok, Thailand. Both patients had underlying diseases. The fungus developed in abscesses with pigmented mycelium at the lower extremity. Cultures from pus and tissue biopsies were positive for dematiaceous fungi. Light microscopic features suggested P. parasitica and which was illustrated by both scanning and transmission electron microscope. The first case was treated with itraconazole with a satisfactory initial response. The second case was successfully treated by surgical removal of the entire lesion.

Cromomicosis: informe de cinco casos con énfasis histopatológico y terapéutico / Chromomycosis: report of five cases with emphasis terapeutic and histopathological

La cromomicosis es una micosis subcutánea y cutánea crónica que predomina en las áreas tropicales y subtropicales. Determinar las características epidemiológicas, clínicas e histopatológicas de los pacientes diagnosticados como cromomicosis; con énfasis en los datos histopatológicos y terapéuticos. Se informan cinco pacientes estudiados entre 1987 y 1997; 3 hombres y dos mujeres con una evolución promedio de 15,8 años. La topografía fue extremidades inferiores y superiores, y un caso diseminada a cara, tronco y extremidades superiores, manifestándose en todos los casos como placas verrucosas. En todos los casos se aisló Fonsecaea pedrosoi. En cuanto a distribución geográfica, 2 pacientes eran de Oaxaca y el resto de Puebla, Tamaulipas y Chiapas respectivamente. En el estudio histopatológico se demostraron cambios proliferativos en la epidermis, constituidos por acantosis e hiperplasia pseudoepiteliomatosa aguda y crónica; así como proliferación vascular y fibrosis: Se pudieron observar células fumagoides tanto dentro de células gigantes multinucleadas, en forma libre, como en los microabscesos de neutrófilos. Las modalidades terapéuticas se basaron en su mayoría en la combinación de procedimientos médicos y quirúrgicos. Debido al número de pacientes incluidos en este estudio, solo podemos describir las características clínicas, histopatológicas y etiología de la cromomicosis. Así mismo podemos concluir que no existe un tratamiento de elección, y que la combinación de procedimientos quirúrgicos y médicos siguen siendo la mejor opción en la cromomicosis

Manejo de la cromomicosis por phialophora verrucosa. (presentacion de un caso)

Se presenta el caso de una micosis subcutanea de 10 años de evolución y múltiples tratamientos fallidos que respondió favorablemente el manejo con INH e intraconazol.

Cromoblastomicose auricular: relato de um caso / Auricular chromoblastomycosis : case report

E apresentado um caso de cromoblastomicose auricular apresentado sob forma de lesao eritemato-escamosa, tendo tido o diagnostico de eczema. Os AA discutem sobre a raridade desta localizacao e aspecto clinico. Embora os poucos casos relatados de cromoblastomicose auricular tenham sido causados pela Fonsecaea pedrosoi, neste o agente etiologico foi a Phialophora verrucosa.

Cromoblastomicose no interior do Estado do Rio Grande do Sul / Chromoblastomycosis in the hinterland of the State of Rio Grande do Sul - Brazil

Foram revistos os protocolos clínico-micológicos de 73 pacientes com cromoblastomicose vistos no interior do Estado do Rio Grande do Sul. A relaçäo entre pacientes homens e mulheres foi de 7:1. Lesöes precoces foram observadas em 8,2% dos pacientes. As lesöes localizavam-se nos membros superiores e face em 8,2 e 4,1% dos doentes, respectivamente. F. pedrosi foi o único agente da infecçäo isolada. Foi discutida a distribuiçäo geográfica peculiar que a micose apresente no Estado

Cromomicose por Fonsecaea pedrosoi apresentando as tres fases conidiais, acrógena, acropleurógena e semi-endógena / Chromomycosis due to Fonsecaea pedrosoi presenting 3 conidial stages: acrogenous, acropleurogenous and semi-endogenous

Una muestra de Fonsecaea pedrosoi (Brumpt) Negroni 1936, fue aislada de un caso de cromomicosis forma plana y presentó tres fases de esporulación: Cladosporium, Rhinocladiella y Phialophora

Feo-hifomicose do couro cabeludo por Phialophora parasitica: registro de um caso / Phalophy-phamycosis of the scalp due to Phialophora parasitica: a case report

Os autores registram caso de feo-hifomicose subcutânea, do couro cabeludo em paciente adulta, do sexo feminino, provocado pela Phialophora parasitica, fungo demácio identificado por AJELLO e cols. Säo descritas as principais características deste hifomiceto. A histopatologia foi conclusiva para o diagnóstico de feo-hifomicose, cujas lesöes simulavam favo do couro cabeludo. Näo havia, no presente caso, manifestaçöes clínicas de uma imunodeficiência

Cromomicosis por Phialophora verrucosa: comunicación del primer caso en México / Chromomycosis by Phialophora verrucosa: report of the first case in Mexico

Se comunica el primer caso en México de cromomicosis producida por Phialophora verrucosa en un paciente de 73 años originario de Caltepec, Puebla, y quien presentaba dos localizaciones del proceso: antebrazo y mano izquierdos y miembro inferior derecho. Se comenta la rareza de casos con dos localizaciones de cromomicosis sobre todo cuando no aparecen simultáneamente y también que es el primer caso en que se demuestra P. verrucosa ya que casi todos los casos mexicanos son producidos por F. pedrosoi. El paciente respondió favorablemente al tratamiento con calciferol, estreptomicina e isoniacida