Human leukocyte antigen class I and MICA haplotypes in a multicase family with Cladophialophora carrionii chromoblastomycosis.

Previous studies carried out in an endemic semiarid region northwest of Venezuela at Falcon State have shown a prevalence of 15.4/1000 of chromoblastomycosis following traumatisms with xenophile vegetation infected with Cladophialophora carrionii. We performed high-resolution DNA typing of human leukocyte antigen (HLA)-A, -B and -C and major histocompatibility complex class I chain related gene A (MICA) alleles and segregation analysis in 49 members of one extended family with 12 affected individuals, who have lived for approximately 70 years in this endemic zone. None of the alleles, haplotypes or genotypes is shared by all the patients. No deviation from the expected HLA haplotype distribution or association of chromoblastomycosis with HLA-A, -B and -C haplotypes was observed. Further, a haplotype-sharing transmission/disequilibria testing of 11 nuclear families did not give enough evidence to claim linkage (P = 0.398), suggesting that genes located in the short arm of chromosome 6 may not be relevant in the immune response toward infection with C. carrionii in this Venezuelan endemic zone. Deleted MICA alleles on HLA-B*4802 haplotypes were present among several members of the extended family, but only two of them were affected.

Chromoblastomycosis in children and adolescents in the endemic area of the Falcón State, Venezuela.

The present paper describes 22 cases of chromoblastomycosis (CBM) caused by Cladophialophora carrionii in children and adolescents (2-19 years old). The patients were seen between 1992 and 2004 and all resided in a CBM endemic area in the semi-arid zone of the Falcón state, Venezuela. Twelve of the 22 patients (54.55%) had close relatives who also had CBM and 19 (86.36%) were male. Lesions consisted of erythematous papules with desquamation or squamous plaques (0.12-14.19 cm in diameter), located primarily on the upper limbs (77.27% of patients). Thirteen of the patients were treated with topical 5-fluorouracil (5-FU; 1% cream), seven with topical ajoene (0.5% gel) and two had electrodesiccation and/or fulguration. Two patients who did not respond to 5-FU were treated with oral itraconazole (100 mg/day for 1 month). Complete clinical and mycological remission was achieved in 17/20 (85%) of the patients treated with 5-FC, ajoene and electrodesiccation and/or fulguration. In addition, similar results were obtained with the two patients who received itraconazole therapy. These cases emphasize the importance of early diagnosis in difficult-to-treat mycotic diseases such as CBM. By early intervention we were able to employ topical treatment with a minimum of adverse effects to achieve a high percentage of favorable therapeutic responses. The patients were thus able to avoid the evolution of the chronic, deforming and incapacitation clinical manifestations associated with CBM.

Saccharomyces cerevisiae en la fabricación del licor cocuy / Sacchromyces cerevisiae in the manufacturing of cocoy liquor

El licor "cocuy" es una bebida artesanal, producida por las comunidades rurales en el Occidente de Venezuela mediante un proceso de fermentación y destilación del mosto extraído del Agave cocui. Este estudio fue enmarcado en el "Programa Agave" con el propósito de contribuir a rescatar esta actividad productiva tradicional. En vista de la falta de información en relación al proceso autóctono se hicieron estudios de las levaduras fermentadoras, la optimización de la producción de etanol y la utilización del residuo de la destilización (vinaza) como medio de cultivo. Los aislados con mayor capacidad fermentativa fueron seleccionados e identificados mediante parámetros morfológicos y metabólicos. Se compararon los niveles de consumo de azúcar de las levaduras con mayor capacidad fermentativa. Se estudió el efecto de la adición del azúcar blanca comercial y/o del fosfato de amonio y en la producción del alcohol en el proceso artesanal. Las concentraciones de azúcares en el mosto se evaluaron por refractometría, y el contenido de alcohol de licor por hidrometría. La utilización de la vinaza para la producción de biomasa como un componente del medio de cultivo fue comparada con un medio sintético mediante medidas del peso seco de la biomasa. Se confirma el papel de sccharomyces ceravisiae en el proceso fermentativo espontáneo. Los resultados in situ evidenciaron un efecto favorable de la elevación del contenido de azúcar (11 a 18 °Brix) y de la adición de fosfato de amonio dibásico (0,2 g/l). En estas condiciones, el tiempo de fermentación del mosto se acortó y la producción de licor aumentó hasta un 92 por ciento. Se demostró la posibilidad de utilizar la vinaza como un componente para un medio de cultivo de esta levadura, para iniciar la fermentación y para la producción de biomasa como fuente de nutrientes de alto valor nutritivo para aves de corral o caprinos. Se recomienda apoyar los esfuerzos para desarrollar de esta importante fuente para los campesinos que habitan las zonas semiáridas de los estados Falcón y Lara

Cladophialophora carrionii: aportes al conocimiento de la endemia en venezuela durante el siglo XX / Cladophialophora carrionii: contributions to the knowledgement of the endemia in Venezuela during the century

Se revisan los aportes en Venezuela al conocimiento de la cromomicosis a partir de mediados del siglo pasado, así como los estudios en relación a los factores de riesgo que podrían explicar la alta prevalencia en el estado Falcón de esta enfermedad rural, laboral y familiar. Se postuló esta micosis profunda como una enfermedad multifactorial. La endemia estaría determinada por la presencia simultánea en un mismo territorio de una población suceptible del agente. CL. carrionii en plantas espinosas; existiendo, además, un alto riesgo de inoculación durante las actividades laborales de los campesinos dedicados a la cría de caprinos en la zona semi-árida. Se señala la tendencia a la endogamia y la presencia del hongo en la reservárea como factores de riesgo determinantes. Se recomienda la búsqueda de los casos entre los familiares de los afectados para su tratamiento precoz, para evitar la invalidez del miembro afectado.

Black fungi: clinical and pathogenic approaches.

Data are presented on the clinically relevant black yeasts and their relatives, i.e., members of the Ascomycete order Chaetothyriales. In order to understand the pathology of these fungi it is essential to know their natural ecological niche. From a relatively low degree of molecular variability of the black yeast Exophiala dermatitidis, potential agent of brain infections in patients from East Asia, it is concluded that this species is an emerging pathogen, currently going through a process of active speciation. It is found to be an oligotrophic fungus in hot, moist environments, such as steambaths. Cladophialophora-, Fonsecaea- and Ramichloridium-like strains, known in humans as agents of chromoblastomycosis, are frequently found on rotten plant material, but the fungal molecular diversity in the environment is much higher than that on the human patient, so that it is difficult to trace the etiological agents of the disease with precision. This approach has been successful with Cladophialophora carrionii, of which cells resembling muriform cells, the tissue form of chromoblastomycosis, were found to occur in drying spines of cacti. Phagocytosis assays provide a method to distinguish between pathogens and non-pathogens, as the killing rates of strict saprobes proved to be consistently higher than of those species frequently known as agents of disease. The therapeutic possibilities for patients with chromoblastomycosis are reviewed.

Cromomisosis por Cladophialophora carrionii: estudio del componente genético en la zona endémica en Venezuela / Chromomycosis for Cladophialophora carrionii: genetic component study in Venezuela endemic zone

Con el fin de conocer los factores que puedan explicar la alta prevalencia de cromomicosis en la zona endémica del estado Falcón, se encuestaron los pobladores de 3 caseríos situados en la zona semiárida. Se elaboraron las genealogías, se aplicó el Indice de Falconer y se ubicó el foco geográfico. Se realizaron estudios micológicos y aislamiento del agente del ambiente peridomiciliari mediante microcultivos. Se diagnosticaron 21 (1,54 por ciento) casos por C. carrionii en 1.356 personas: 20 (95 por ciento) pertenecían a un mismo grupo familiar, con una relación de uniones consanguíneas de 1/4; 7 (54 por ciento) de los casos producto directo de éstas. Se demostró la presencia del agente en una cactácea, Opuntia caribaea (guazábara). Se observó un patrón de herencia correspondiente a un gen autosómico recesivo. Se estimó una heredabilidad del 47 por ciento (5,6 por ciento de hermanos afectados). Nuestros resultados demuestran una alta participación del componente genético en esta micosis profunda, factor determinante para explicar la endemia en estas poblaciones rurales, sedentarias y aisladas con tendencia a las uniones consanguíneas. Se proponen estrategias racionales para el control de la endemia

[Chromomycosis by Rhinocladiella aquaspera: The first case in Venezuela.]. / Cromomicosis por Rhinocladiella aquaspersa: descripción del primer caso en Venezuela.

A case of chromomycosis induced by Rhinocladiella aquaspersa is decribed in Falcon State, which is considered as the first case in Venezuela, and the fifth one in the whole world of this species of Rhinocladiella. The case was observed in a five year-old child from the south part of Falcon State. The child presented wounds of two years of duration, in his upper left limb. These wounds were characterized by queloide cicatrices, and contiguous nodules, with a scalyo-scab surface. The disease was diagnosed through direct observation of cells extracted from the wounds and cleared with potassium hydroxide at 10%. The identification of R. aquaspersa was made by 1) The isolation in culture, using Lactritmel, Sabouraud-agar and Sablac as culture media; and 2) By observation, in slide culture, of dissemination structures characterized by erect conidiophores which were either simple or branched. The thermo-tolerance test applied indicate an optimum development temperature of 28 degrees C. It was not possible to reproduce the illness in Balb/c mice. The patient showed an unsatisfactory response to Itraconazol medication and improvement to an external local application therapy. Up to the present time, 370 cases of chromomycosis have been studied in Falcon State, with 21 cases in children. The suggests the existence of a susceptibility to this kind of mycosis from childhood.

[Mycetoma: report of 3 cases in Falcón State, Venezuela]. / Micetoma: reporte de tres casos en el Estado Falcón, Venezuela.

This is a report of three cases of mycetome studied in our laboratory during the last four years. We also informed about the evolution of the first case produced by Pyrenochaeta romeroi nova species reported by Borelli in 1959. All the patients were farmer men, 18, 42 and 54 years old, from the semiarid and subhumid zones of Falcon State, located on the northwest region of Venezuela. Clinically, all patients presented the mycetome syndrome (subcutaneous edema, sinus tract and "granules"), with a evolution of six months to three years. The lesions were localized in the inferior limbs in two of the patients and in the thorax in one of them. The clinical diagnose was confirmed by microbiological and mycological studies, isolating Nocardia brasiliensis in two cases and Madurella grisea in one. Two patients treated with Sulfas showed remission of the disease in a two years follow up. Including this report, 15 cases of mycetome have been reported in the Falcon State (Venezuela) in a period of thirty five years, representing only 9, 14% of the national casuistic. Actinomadura madurae and N. brasiliensis have been the most frequently isolated agents in this region. In order to determine the real endemicity of the mycetome in Venezuela, multidisciplinary clinical, inmunological, microbiological and epidemiological studies are recommended. According to the reported casuistic (15 cases), the mycetome should be a low frequency disease in Falcon State, Venezuela. We confirm that early diagnosis favors an efficient therapy.

Virulence and pathogenicity of human and environmental isolates of Cladosporium carrionii in new born ddY mice.

Three strains of Cladosporium carrionii, two human isolates and one from a xerophilous plant, were used to study the effect of culture conditions in 106 newborn ddY mice. Growth in a complex medium (YPG) and a basal synthetic medium (BSM) was compared. Filamentous forms developed during static incubation while conidia were readily formed with shaking. Mice inoculated intraperitoneally were sacrificed and autopsied after 4 weeks. Mortality was related only to sporulated exponential phase growing cells. Invasiveness ability was preserved in all experimental conditions. BSM medium that inhibited exopigment formation appeared more suitable than YPG to obtain intact cells for further studies. Biochemical and physiological alteration associated with shape changes during differentiation of vegetative cells into spores could play an important role in virulence of C. carrionii.

Itraconazol: ensayo terapéutico en micosis profundas y leishmaniasis / Itraconazole: therapeutic assay in deep mycoses and leishmaniasis

Hemos administrado per os este imidazólico (Janssen R 51 211) a 2 pacientes con esporotricosis, 10 con paracoccidioidosis, 3 con micetomas, 14 con cromomicosis, 5 con leishmaniasis. Los controles clínicos y de laboratorio mostraron tolerancia completa, ausencia de efectos colaterales, actividad satisfactoria en paracoccidioidosis, esporotricosis y cromomicosis por Cladosporium carrionii; actividad alentadora en un micetoma por Madurella grisea y en algunos casos de Leishmaniasis, incluyendo uno por Leishmania mexicana pifanoi. La dosis diaria empleada ha sido de 50 mg. en paracoccidioidosis, 200 ó 400 mg. en cromomicosis, 200 en micetomas, esporotricosis y leishmaniasis

Liberation of protoplasts from the yeast-like form of Paracoccidioides brasiliensis by enzymatic treatment.

Spherical and osmotically sensitive protoplasts were released from cultures of the yeast-like form of Paracoccidioides basilienisis strain IVIC Pb9 through the action of a mixture of crude enzyme preparations: alpha and beta-glucanases and chitinase, obtained from culture filtrates of Cladosporium resinae, Basidiomycete QM 806 and Streptomyces sp respectively. The highest efficiency of protoplast liberation was achieved when each crude enzyme preparation was used at 1 mg/ml.

Histoplasma capsulatum: chemical variability of the yeast cell wall.

Four strains of Histoplasma capsulatum were analyzed to observe any variations in their cell wall composition. Strain G-184B produced the same amount of sugars reported by Domer (2) although qualitative analyses agreed with Kanetsuna's report (4) as far as the presence of glucose, galactose, and mannose was concerned. Our results agree with previous reports in classifying H. capsulatum into chemotypes I and II, the latter having more alpha-glucan in its cell wall. A decrease in the amount of fraction 2 (alpha-glucan) of strain G-184B was observed when the strain was subcultured in vitro for many years, an effect similar to that reported for Paracoccidioides brasiliensis (12, 13).

Effects of cyanein and ramihyphin A on the dimorphism of Paracoccidioides brasiliensis.

As a preliminary step in the study of dimorphism in Paracoccidioides brasiliensis, the effects of cyanein and ramihyphin A were studied. These antibiotics have been reported to induce morphological changes in fungi. The results obtained suggest that ramihyphin A induce swelling of hyphae while partially inhibiting Y leads to M transformation with the production of an incipient and swollen mycelium. Cyanein did not affect the mycelial morphology as did ramihyphin A. However, the Y leads to M transformation was inhibited and, also, the M leads to Y transformation was blocked with the production of a few yeast cells which were not released from the mycelium.