[Comparison of different methods of DNA extraction from blood to detect fungal DNA by PCR]. / Comparación de métodos de extracción de ADN de sangre para detectar ADN fúngico por PCR.

Comparison of different methods of DNA extraction from blood to detect fungal DNA by PCR. Invasive fungal infections (IFI) are associated with high mortality by reaching levels of 50%, and also with a significant failure in antifungical treatments. This fact mostly obeys to difficulties in obtaining a fast and accurate mycologic diagnosis due to the low sensitivity of conventional methods, mainly in neutropenic and AIDS patients. Various methods based on fungal DNA study are currently being used for the diagnosis of mycotic infections. We herein evaluated two procedures of extraction and purification of fungal DNA in blood for their use in PCR detection. Both of them showed equal efficiency in obtaining high performance DNA with universal primers ITS land ITS 4 as target.

Comparación de métodos de extracción de ADN de sangre para detectar ADN fúngico por PCR / Comparison of different methods of DNA extraction from blood to detect fungal DNA by PCR

La infección fúngica invasora (IFI) está asociada a un alto índice de mortalidad, que alcanza el 50% debido a la frecuente falla en el tratamiento antifúngico. Existen dificultades para realizar un diagnóstico micológico rápido y certero dada la baja sensibilidad de los métodos convencionales, especialmente en pacientes neutropénicos y con SIDA. Numerosos métodos para diagnosticar infecciones micóticas basados en el estudio del ADN fúngico están actualmente en desarrollo. Nosotros evaluamos la utilidad de dos procedimientos de extracción y purificación del ADN fúngico presente en sangre para su posterior detección por PCR. Ambos métodos resultaron igualmente eficientes para obtener ADNs de óptima calidad y para realizar la técnica de PCR con los iniciadores universales para hongos ITS 1 e ITS 4.

Invasive fungal infections (IFI) are associated with high mortality by reaching levels of 50%, and also with a significant failure in antifungical treatments. This fact mostly obeys to difficulties in obtaining a fast and accurate mycologic diagnosis due to the low sensitivity of conventional methods, mainly in neutropenic and AIDS patients. Various methods based on fungal DNA study are currently being used for the diagnosis of mycotic infections. We herein evaluated two procedures of extraction and purification of fungal DNA in blood for their use in PCR detection. Both of them showed equal efficiency in obtaining high performance DNA with universal primers ITS 1and ITS 4 as target.

[Prevalence of Candida albicans and Candida non-albicans in clinical samples during 1999-2001]. / Prevalencia de Candida albicans y Candida no albicans en diferentes muestras clínicas. Período 1999-2001.

The importance of epidemiological monitoring of yeasts involved in pathologic processes is unquestionable due to the increase of these infections over the last decade, the changes observed in species causing candidiasis, and empirical antifungal treatment. At the Mycology Center, 1006 isolates from a wide range of clinical samples were studied during 1999-2001. Candida albicans (40.3%) was the most isolated species, although, the Candida no albicans species with 54.9% showed the major prevalence. In blood cultures Candida parapsilosis (34.9%), C. albicans (30.2%) and C. tropicalis (25.6%) were recovered most frequently while C. glabrata represented only 2.3%. C. albicans with 60%-80% was the predominant specie in mucosal surface. We also detected Candida mediastinistis, which alert us over the importance at this location. Urinary tract infections caused by yeasts were more frequent in hospitalized patients, being C. albicans (47.7%), the most commonly isolated, followed by C. glabrata (24.8%) and C. tropicalis (20.0%). In the candidal onychomycoses, C. parapsilosis (37.7%) outplaced C. albicans (22.0%). Fluconazole susceptibility studies of Candida species allowed us to conclude that the majority of C. albicans islolates are susceptible, and that the highest resistance averages were observed in C. glabrata (21.41%) and C. krusei (69.23%).

Prevalencia de Candida albicans y Candida no albicans en diferentes muestras clínicas: Período 1999-2001 / Prevalence of Candida albicansand Candida no albicans inclinical samples during 1999-2001

Las levaduras implicadas en procesos patológicos son de indiscutible importancia debido al incremento experimentado por estas infecciones en las últimas décadas, a los cambios observados en las especies causales y al uso empírico de antifúngicos. En el Centro de Micología se estudiaron 1006 aislamientos provenientes de una amplia gama de muestras clínicas durante el periodo 1999-2001. Candida albicans con 40,3% resultó la especie de mayor frecuencia de aislamiento, pero las especies de Candida no albicans con 54,9% resultaron de mayor prevalencia y el 4,8% fueron otras levaduras. En los hemocultivos Candida parapsilosis con 34,9%, C. albicans con 30,2% y C. tropicalis con 25,6% resultaron las más recuperadas, mientras que C. glabrata se presentó con un 2,3%. En las secreciones mucosas C.albicans con 60%-80% fue la especie preponderante. Hemos detectado especies de Candida causantes de mediastinitis, lo que nos alerta sobre su importancia en estos procesos. Las infecciones del tracto urinario por levaduras se detectaron en mayor frecuencia en individuos hospitalizados, resultando C. albicans con 47,7% la especie más aislada, y dentro de Candida no albicans, C. glabrata con 24,8% y C. tropicalis con 20,0%. En las onixis candidiásicas C. parapsilosis con 37,7% desplazó a C.albicans con 22,0% de este lugar anatómico. Los estudios de sensiblidad al fluconazol de las especies de Candida nos permiten concluir que C.albicans es una especie sensible y que los mayores porcentajes de resistencia se observaron en C. glabrata (21,41%) y and C. krusei (69,23%).

The importance of epidemiological monitoring of yeasts involved in pathologic processes is unquestionable due to the increase of these infections over the last decade, the changes observed in species causing candidiasis, and empirical antifungal treatment. At the Mycology Center, 1006 isolates from a wide range of clinical samples were studied during 1999-2001. Candida albicans (40.3%) was the most isolated species, although, the Candida no albicans species with 54.9% showed the major prevalence. In blood cultures Candida parapsilosis (34.9%), C. albicans (30.2%) and C. tropicalis (25.6%) were recovered most frequently while C. glabrata represented only 2.3%. C. albicans with 60%-80% was the predominant specie in mucosal surface. We also detected Candida mediastinistis, which alert us over the importance at this location. Urinary tract infections caused by yeasts were more frequent in hospitalized patients, being C. albicans (47.7%), the most commonly isolated, followed by C. glabrata (24.8%) and C. tropicalis (20.0%). In the candidal onychomycoses, C. parapsilosis (37.7%) outplaced C. albicans (22.0%). Fluconazole susceptibility studies of Candida species allowed us to conclude that the majority of C. albicans islolates are susceptible, and that the highest resistance averages were observed in C. glabrata (21.41%) and C. krusei (69.23%).

Prevalencia de Candida albicans y Candida no albicans en diferentes muestras clínicas. Período 1999-2001. / [Prevalence of Candida albicans and Candida non-albicans in clinical samples during 1999-2001]

The importance of epidemiological monitoring of yeasts involved in pathologic processes is unquestionable due to the increase of these infections over the last decade, the changes observed in species causing candidiasis, and empirical antifungal treatment. At the Mycology Center, 1006 isolates from a wide range of clinical samples were studied during 1999-2001. Candida albicans (40.3

) was the most isolated species, although, the Candida no albicans species with 54.9

showed the major prevalence. In blood cultures Candida parapsilosis (34.9

), C. albicans (30.2

) and C. tropicalis (25.6

) were recovered most frequently while C. glabrata represented only 2.3

. C. albicans with 60

-80

was the predominant specie in mucosal surface. We also detected Candida mediastinistis, which alert us over the importance at this location. Urinary tract infections caused by yeasts were more frequent in hospitalized patients, being C. albicans (47.7

), the most commonly isolated, followed by C. glabrata (24.8

) and C. tropicalis (20.0

). In the candidal onychomycoses, C. parapsilosis (37.7

) outplaced C. albicans (22.0

). Fluconazole susceptibility studies of Candida species allowed us to conclude that the majority of C. albicans islolates are susceptible, and that the highest resistance averages were observed in C. glabrata (21.41

) and C. krusei (69.23

).

Susceptibility of Candida species to fluconazole assayed by a disk diffusion method with automated reading versus a microdilution method.

In vitro susceptibilities of 290 isolates of Candida spp to fluconazole were evaluated by a new NCCLS M44-P agar disk diffusion method that was read and interpreted automatically by the BIOMIC image-analysis plate reader system. Disk test results were compared to results obtained by a modified NCCLS M27-A broth microdilution method using RPMI-1640 supplemented with 2% dextrose. Overall agreement between both methods was 90.0%. Category agreement between the broth and disk test results for susceptible, susceptible dose-dependent and resistant disk results were 99.6%, 19.05% and 52.17%, respectively. No very major discrepancies, 1.03% major discrepancies, and 8.97% minor discrepancies were observed between results of the two test methods. This analysis suggests that results from both methods correlate highly for Candida strains susceptible to fluconazole. The lower agreement between the two methods for resistant and susceptible dose-dependent isolates was due to strains near or on that breakpoint, different media (Mueller-Hinton vs RPMI), agar vs broth, 80% vs 50% endpoints, and trailing growth near the endpoints.

Susceptibility of Candida species to fluconazole assayed by a disk diffusion method with automated reading versus a microdilution method

In vitro susceptibilities of 290 isolates of Candida spp to fluconazole were evaluated by a new NCCLS M44-P agar disk diffusion method that was read and interpreted automatically by the BIOMIC image-analysis plate reader system. Disk test results were compared to results obtained by a modified NCCLS M27-A broth microdilution method using RPMI-1640 supplemented with 2

. Category agreement between the broth and disk test results for susceptible, susceptible dose-dependent and resistant disk results were 99.6

major discrepancies, and 8.97

minor discrepancies were observed between results of the two test methods. This analysis suggests that results from both methods correlate highly for Candida strains susceptible to fluconazole. The lower agreement between the two methods for resistant and susceptible dose-dependent isolates was due to strains near or on that breakpoint, different media (Mueller-Hinton vs RPMI), agar vs broth, 80

endpoints, and trailing growth near the endpoints.

Susceptibility of Candida species to fluconazole assayed by a disk diffusion method with automated reading versus a microdilution method.

In vitro susceptibilities of 290 isolates of Candida spp to fluconazole were evaluated by a new NCCLS M44-P agar disk diffusion method that was read and interpreted automatically by the BIOMIC image-analysis plate reader system. Disk test results were compared to results obtained by a modified NCCLS M27-A broth microdilution method using RPMI-1640 supplemented with 2

dextrose. Overall agreement between both methods was 90.0

. Category agreement between the broth and disk test results for susceptible, susceptible dose-dependent and resistant disk results were 99.6

, 19.05

and 52.17

, respectively. No very major discrepancies, 1.03

major discrepancies, and 8.97

minor discrepancies were observed between results of the two test methods. This analysis suggests that results from both methods correlate highly for Candida strains susceptible to fluconazole. The lower agreement between the two methods for resistant and susceptible dose-dependent isolates was due to strains near or on that breakpoint, different media (Mueller-Hinton vs RPMI), agar vs broth, 80

vs 50

endpoints, and trailing growth near the endpoints.

Experimental coccidioidomycosis in hamsters. Disease kinetics and death curve in relation to infective dose.

A study of experimental coccidioidomycosis in hamsters (Mesocricetus auratus) is presented. Two experiments were conducted on 75 animals inoculated intracardially with the mycelial form of Coccidioides immitis. The first research (experiment I) studied the kinetics of experimental disease in 15 hamsters inoculated with 300 C. immitis arthroconidia. The parameters studied were: (a) presence of macroscopic lesions in the brain, lungs, liver, spleen and kidneys; (b) microscopic identification of spherules in wet mount preparations of these specimens; (c) samples from all organs cultured at 37 degrees C on Sabouraud glucose agar; (d) blood cultures drawn every 24 h during the first week and subsequently every 48 h and (e) histopathological studies of all organs. The second experiment (experiment II) determined the relationship between the inoculum size and death curve in six groups of 10 animals each, which had received doses of 10, 50, 100, 150, 200 and 300 arthroconidia, respectively. On day 14 post-inoculation, all the animals underwent skin tests and 1 ml of blood was obtained by cardiac puncture to detect antibodies. Disseminated disease with persistent fungaemia developed in all the studied animals. Coccidioides immitis was recovered from all organs, with the lungs being the first to present disease. Death occurred in all groups, regardless of the dose of arthroconidia and 83.3% died between day 22 and day 28 post-infection. The use of this model is proposed for the biological standardization of antigens, the study of prophylactic measures and the "in vivo" evaluation of new antifungal treatments.

New advances in antifungal treatment.

Clinical aspects of treatment of invasive aspergillosis, infections caused by dematiaceous fungi, and mycoses caused by endemic, dimorphic fungi, are described in this review.

Histopathological evolution of experimental paracoccidioidomycosis in Wistar rats.

Twenty Wistar rats were inoculated, by the intracardiac route, with 0.5 ml each of a yeast phase suspension of Paracoccidioides brasiliensis Utero strain. The rats were sacrificed at regular intervals post-infection, at which time their lungs, heart, liver, spleen and kidneys were removed, fixed and stained for study. The parameters of interest for the lung specimens were: (a) extent of the lesions; (b) number of fungi; (c) presence of a lymphomononuclear halo. Extrapulmonary lesions were also sought. Until the fourth month post-infection, the lesions were progressive in nature, contained great numbers of viable fungi, and were surrounded by an important lymphomononuclear halo which tended to be confluent. At four and a half months p.i., the extent of the pulmonary lesions was reduced, the granulomas were less compact with fewer viable fungi, macrophages showed microvacuolation, and the lymphomononuclear halo was less pronounced. Extrapulmonary lesions, which were frequently identified in the first months post-infection, diminished from the seventh month onwards. The histological characteristics of extrapulmonary lesions were always the same as those found in the lungs. Infection tended to be controlled by the animals from the fourth month, but without complete resolution of the lesions.

Estudio comparativo de la patogenicidad y la antigenicidad de 6 cepas de Paracoccidioides brasiliensis / Comparative study of pathogenicity and antigenicity of six Paracoccidioides brasiliensis strains

Fueron estudiadas en forma comparativa 6 cepas de P. brasiliensis con el proposito de determinar su patogenicidad para la rata y su antigenicidad. Las mismas fueron aisladas de: 1) biopsia de cuello uterino en 1989 (U), 2) biopsia de mucosa bucal en 1988 (V), 3) aspiracion osea en 1991 (63265), 4) testiculo de cobayo 1984 (C24), 5) puncion-aspiracion ganglionar en 1986 (G) y 6) cepa proveniente de la Escola Paulista de Medicina (339). Se prepararon antigenos citoplasmaticos liofilizados de cada una de ellas, en la concentracion final de 100mg/ml y se realizaron pruebas de inmunodifusion frente a 6 sueros patrones positivos de ratas....

[Comparative study of the pathogenicity and antigenicity of 6 Paracoccidioides brasiliensis strains]. / Estudio comparativo de la patogenicidad y la antigenicidad de 6 cepas de Paracoccidioides brasiliensis.

A comparative study of antigenicity and pathogenicity for rats of six Paracoccidioides brasiliensis strains was carried out. The antigenic capacity "in vitro" of cytoplasmic extract from each strain was determined by immunodiffusion test against 6 serum samples obtained from rats experimentally infected with Paracoccidioides brasiliensis, that had presented positive reactions with a metabolic control antigen. The cytoplasmic extracts were used at final concentration of 100 mg/ml. All of them showed 2 or 3 precipitation bands in this assay. One hundred twenty Wistar rats both sexes weighing approximately 200 g, were inoculated intracardiacally with suspensions of the yeast phase of different P. brasiliensis strains. Two concentrations containing 3 x 10(7) and 5 x 10(7) cells/ml of each isolate were prepared. The inoculated animals were divided in two groups, one was left to its spontaneous outcome and the percentages of deaths were registered and the other rats were sacrificed at 14, 28, 56 and 70 days post-infection. The following parameters were taken into account for evaluation: A) presence of macroscopic granulomas in lung, liver, spleen and kidney; B) presence of P. brasiliensis in microscopic exams of the same organs, in wet preparations and in histologic sections stained by H&E; C) culture of lung and D) immunodiffusion test using pre-mortem serum samples and the homologous antigen. The correlation between the most important parameters studied in each strain are summarized as follow: As no significant differences between the two inocula employed for each strain was observed, the before mention results are the average of those obtained with each inoculation doses.(ABSTRACT TRUNCATED AT 250 WORDS)

Tratamiento de la coccidioidomicosis experimental en ratas Wistar / Experimental treatment of coccidioidomycosis in Wistar rats

Se realizó un estudio comparativo entre Itraconazol (I) y Fluconazol (F) en el tratamiento de la coccidioidomicosis experimental en ratas Wistar. Treinta animales de 250 g de peso fueron inoculados con 200 artroconidias de Coccidioides immitis por vía intercardíaca. Ambas drogas fueron administradas por gastroclisis a razón de 200 mg//kg/día durante 5 semanas a partir del 7 día post-inoculación. El grupo control recibió el solvente de las drogas. Los animales que sobrevivieron 2 semanas después de concluído el tratamiento fueron sacrificados y se le realizaron los siguientes estudios: 1) examen macroscópico (presencia de granulomas) y microscópico (presencia de esporangios) en los pulmones; 2) unidades formadoras de colonias en homogeneizado de pulmones; 3) estudio histopatológico de pulmones con H & E realizando una evaluación semi-cuantitativa contando: a) número de granulomas por campo de 50x; b) número de esporangios con esporangiosporos en cada granuloma y c) número de esporangios vacíos. Los resultados obtenidos mostraron que el itraconazol fue más eficaz que el fluconazol en el tratamiento de este modelo experimental, en la evaluación efectuada a través de varios de los parámetros considerados: 1) se comprobaron menor cantidad de cultivos pulmonares positivos, (30 por ciento vs 100 por ciento); 2) menor número de UFC/g de pulmón (<1x10 vs 1.7x10 ); 3) menos cantidad de granulomas pulmonares en el estudio histopatológico (1/14 campos vs 1/4 campos); 4) menor cantidad de esporangios por granuloma (0 vs 33) y 5) mayor número de esporangios vacíos o alterados. Cabe señalar, sin embargo, que el fluconazol fue capaz de reducir el número de UFC/g pulmón,comparando con el grupo control

Tratamiento de la coccidioidomicosis experimental en ratas Wistar / Experimental treatment of coccidioidomycosis in Wistar rats

Se realizó un estudio comparativo entre Itraconazol (I) y Fluconazol (F) en el tratamiento de la coccidioidomicosis experimental en ratas Wistar. Treinta animales de 250 g de peso fueron inoculados con 200 artroconidias de Coccidioides immitis por vía intercardíaca. Ambas drogas fueron administradas por gastroclisis a razón de 200 mg//kg/día durante 5 semanas a partir del 7 día post-inoculación. El grupo control recibió el solvente de las drogas. Los animales que sobrevivieron 2 semanas después de concluído el tratamiento fueron sacrificados y se le realizaron los siguientes estudios: 1) examen macroscópico (presencia de granulomas) y microscópico (presencia de esporangios) en los pulmones; 2) unidades formadoras de colonias en homogeneizado de pulmones; 3) estudio histopatológico de pulmones con H & E realizando una evaluación semi-cuantitativa contando: a) número de granulomas por campo de 50x; b) número de esporangios con esporangiosporos en cada granuloma y c) número de esporangios vacíos. Los resultados obtenidos mostraron que el itraconazol fue más eficaz que el fluconazol en el tratamiento de este modelo experimental, en la evaluación efectuada a través de varios de los parámetros considerados: 1) se comprobaron menor cantidad de cultivos pulmonares positivos, (30 por ciento vs 100 por ciento); 2) menor número de UFC/g de pulmón (<1x10 vs 1.7x10 ); 3) menos cantidad de granulomas pulmonares en el estudio histopatológico (1/14 campos vs 1/4 campos); 4) menor cantidad de esporangios por granuloma (0 vs 33) y 5) mayor número de esporangios vacíos o alterados. Cabe señalar, sin embargo, que el fluconazol fue capaz de reducir el número de UFC/g pulmón,comparando con el grupo control

Treatment of experimental cryptococcosis with SCH 39304 and fluconazole.

The efficacy of two triazoles, SCH 39304 and fluconazole, in the treatment of disseminated cryptococcosis in Wistar rats was determined. A total of 160 rats were inoculated intracardiacally with 2 x 10(5) cells of Cryptococcus neoformans. Both drugs were administered by gavage once daily, at three doses (8, 16, and 32 mg/kg/day). Two treatment schedules were followed: (i) treatment began 1 week after infection and continued for 3 weeks and (ii) prophylaxis treatment began 3 days before infection and continued an additional 3 weeks. Evaluation was based on (i) macroscopic examination of lungs, (ii) microscopic examination of brains and lungs, (iii) histopathology of brains and lungs, and (iv) determination of number of CFU in brains. The number of CFU was the best measure of activity. SCH 39304 was more active than fluconazole in both regimens, and, prophylactically, SCH 39304 was able to achieve biological cures.

Compuestos triazólicos en el tratamiento de la histoplasmosis diseminada experimental del hamster / Triazoles compound in the treatment of disseminated experimental histoplasmosis in hamsters

Se llevó a cabo un estudio comparativo de la eficacia de cuatro compuestos triazólicos en el tratamiento de la histoplasmosis experimental del hamster. Fueron utilizados en total 110 hamsters, de ambos sexos. Estos animales se inocularon por vía intracardíaca con una suspensión de la fase levaduriforme del Histoplasma capsulatum. Los tratamientos comenzaron 1 semana después de la infección y se prolongaron por 3 semanas. Todas las drogas fueron administradas por gastroclisis una vez por día, a las dosis de 8 y 16 mg/kg de peso. Tres grupos de hamsters fueron empleados como control y recibieron los solventes de los antifúngicos estudiados. Fueron administrados los siguientes triazoles: itraconazol, fluconazol, Sch 39.304 y Bay r 3783. Todos los animales se sacrificaron una semana después de terminados los tratamientos. La evaluación de los resultados se realizó en base a los siguientes criterios: 1)Aspecto macroscópico de higado y bazo, 2)examen microscópico de frotis teñidos con Giemsa de los mismos órganos, 3)cortes histológicos de bazo teñidos por el método de P.A.S.y 4)cultivos de bazo (cultivos masivos de un homogeneizado del órgano y determinación de U.F.C/g). El itraconazol y el Sch 39.304 redujeron en forma muy marcada las alteraciones macroscópicas, la positividad de los exámenes microscópicos y los cultivos. Los animales tratados con fluconazol y con Bay r 3783 a la dosis de 16 mg/kg/día, sólo mostraron escasa reducción del número de U.F.C/g de bazo, en relación a los controles. El itraconazol es una droga de probada eficacia en la histoplasmosis humana. El Sch 39.304 parece ser un antifúngico interesante en el tratamiento de esta enfermedad, en especial si se tienen en cuenta sus propiedades fármaco-cinéticas distintas de las itraconazol lo que facilitaría su aplicación en otras localizaciones de la histoplasmosis diseminada

Criptococosis humana producida por una variante puntiforme de cryptococcus neoformans / Cryptococcosis human produced for an variant punctiform of cryptococcus neoformans

Se estudiaron la patogenicidad y la susceptibilidad antifúngica "in vitro" e "in vivo" de una cepa puntiforme de Cryptococcus neoformans var. neoformans, aislada del LCR de un paciente parkinsoniano.Sembrado el material en los medios habituales, se observó desarrollo, tras 20 días de incubación, de colonias pequeñas, de menos de 1mm de diámetro. No se comprobaron diferencias significativas en la patogenicidad de esta cepa con respecto a otra utilizada como control. Si bien la CIM para la anfotericina B fue elevada, esta resistencia no fue evidenciada en el modelo experimental del ratón

Compuestos triazólicos en el tratamiento de la histoplasmosis diseminada experimental del hamster / Triazoles compound in the treatment of disseminated experimental histoplasmosis in hamsters

Se llevó a cabo un estudio comparativo de la eficacia de cuatro compuestos triazólicos en el tratamiento de la histoplasmosis experimental del hamster. Fueron utilizados en total 110 hamsters, de ambos sexos. Estos animales se inocularon por vía intracardíaca con una suspensión de la fase levaduriforme del Histoplasma capsulatum. Los tratamientos comenzaron 1 semana después de la infección y se prolongaron por 3 semanas. Todas las drogas fueron administradas por gastroclisis una vez por día, a las dosis de 8 y 16 mg/kg de peso. Tres grupos de hamsters fueron empleados como control y recibieron los solventes de los antifúngicos estudiados. Fueron administrados los siguientes triazoles: itraconazol, fluconazol, Sch 39.304 y Bay r 3783. Todos los animales se sacrificaron una semana después de terminados los tratamientos. La evaluación de los resultados se realizó en base a los siguientes criterios: 1)Aspecto macroscópico de higado y bazo, 2)examen microscópico de frotis teñidos con Giemsa de los mismos órganos, 3)cortes histológicos de bazo teñidos por el método de P.A.S.y 4)cultivos de bazo (cultivos masivos de un homogeneizado del órgano y determinación de U.F.C/g). El itraconazol y el Sch 39.304 redujeron en forma muy marcada las alteraciones macroscópicas, la positividad de los exámenes microscópicos y los cultivos. Los animales tratados con fluconazol y con Bay r 3783 a la dosis de 16 mg/kg/día, sólo mostraron escasa reducción del número de U.F.C/g de bazo, en relación a los controles. El itraconazol es una droga de probada eficacia en la histoplasmosis humana. El Sch 39.304 parece ser un antifúngico interesante en el tratamiento de esta enfermedad, en especial si se tienen en cuenta sus propiedades fármaco-cinéticas distintas de las itraconazol lo que facilitaría su aplicación en otras localizaciones de la histoplasmosis diseminada

Criptococosis humana producida por una variante puntiforme de cryptococcus neoformans / Cryptococcosis human produced for an variant punctiform of cryptococcus neoformans

Se estudiaron la patogenicidad y la susceptibilidad antifúngica "in vitro" e "in vivo" de una cepa puntiforme de Cryptococcus neoformans var. neoformans, aislada del LCR de un paciente parkinsoniano.Sembrado el material en los medios habituales, se observó desarrollo, tras 20 días de incubación, de colonias pequeñas, de menos de 1mm de diámetro. No se comprobaron diferencias significativas en la patogenicidad de esta cepa con respecto a otra utilizada como control. Si bien la CIM para la anfotericina B fue elevada, esta resistencia no fue evidenciada en el modelo experimental del ratón