Search

Disseminated histoplasmosis is the most serious form of the disease produced by the fungus Histoplasma capsulatum. Histoplasmosis was first described by Darling in 1906, and it is endemic in certain areas of Central and Southeastern regions of USA, and most Latin America countries, including the Pampa húmeda and Mesopotamia regions in Argentina, but in other continents it has a limited regional distribution. Lung involvement predominates in 90% of cases, but H. capsulatum may involve bone and soft tissues. Bone lesions without other signs of infection are very rare, and are often misdiagnosed as cancer. We report a case of disseminated histoplasmosis in a man with a myelodysplasia who presented a left tibial abscess, without any clinical evidence of pulmonary involvement. The patient was successfully treated with itraconazole.

The treatment of mycetomas varies according to their etiological agents and the clinical state of the patient. For the treatment of eumycetomas, the azole derivatives are the drugs of choice, with itraconazole rendering better results than ketoconazole and presenting better tolerance. Actinomycetomas are treated according to different therapeutic schemes: dapsone plus sulfamethoxazol-trimethoprim (SMT), and streptomycin or amikacin or amoxicillin plus clavulanic acid. The first therapeutic scheme is very useful in the treatment of Nocardia mycetoma, while the association of amikacin plus SMT is the best treatment for those cases produced by Actinomadura madurae. Ciprofloxacin is a very useful drug for the treatment of actinomycotic mycetomas with bone lesions. Although there are several criteria for evaluating clinical outcome there is no accepted criterion of cure. During the 1990s, there was a remarkable increase in the incidence of coccidioidomycosis in California, USA. An almost ten-fold increase in the number of cases was registered during 1992 and 1993 over the usual incidence. A gradual reduction in coccidioidomycosis cases was observed in the late 1990s. This particular coccidioidomycosis outbreak took place in areas of low endemicity, as well as in those of usual high endemicity. Among the factors believed to have influenced this phenomenon were a drought followed by abundant winter/spring rainfall, increased immigration of susceptible individuals, increase in excavation/construction work and a better diagnosis of the infection, particularly in the last part of the decade. The majority of patients presented the usual clinical manifestations of symptomatic primary infection but an unusual number of cases with acute respiratory failure were observed.

Despite advances in diagnosis and treatment, the epidemiological status of the human immunodeficiency virus (HIV) infection is far from under control in most of the developing world. Sub-Saharan Africa, Southeast Asia and India show increased rates of new infections. In Latin America and the Caribbean there were 1.6 million estimated cases of HIV-infected patients at the end of 1997. Fungal diseases have been one of the most relevant diagnoses in relation to the acquired immunodeficiency syndrome (AIDS). Infections due to Candida species and Cryptococcus neoformans var. neoformans are common worldwide. Histoplasma capsulatum, Coccidioides immitis and Penicillium marneffei are important causes of disease in endemic areas. Infection due to Sporothrix schenckii, Blastomyces dermatitidis and Paracoccidioides brasiliensis are uncommon even where they are endemic. Phaeohyphomycetes, hyalohyphomycetes and zygomycetes are still rare as a cause of disease among AIDS patients. However, agents pertaining to these groups, such as Aspergillus spp., have an increasing incidence. Superficial mycoses due to dermatophytes have special features from epidemiological, clinical and therapeutic points of view.

Summary 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 °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.

Internal transcribed spacer (ITS) genes including the 5.8S ribosomal (r)RNA of Paracoccidioides brasiliensis were amplified and the DNA sequences were determined. Based on a comparison of the sequence information, a new polymerase chain reaction (PCR) primer pair was designed for specific amplification of DNA for P. brasiliensis. This primer pair amplified a 418-bp DNA sequence and was 100% successful in identifying 29 strains of P. brasiliensis (including the reference strains) isolated from the regions of Brazil, Costa Rica, Japan, Argentina or from different sources. The results of specificity tests of these primers to compare the fungus with those of Aspergillus fumigatus, Blastomyces dermatitidis, Candida albicans, Cryptococcus neoformans, Histoplasma capsulatum and Penicillium marneffei are also reported.

Restriction fragment length polymorphism (RFLP) of mitochondrial DNA (mtDNA) from 49 clinical Fonsecaea pedrosoi isolates (18 isolates from Japan, 17 from Madagascar, 7 from Argentina, 5 from Venezuela, 1 from Costa Rica and 1 unknown) was studied. The 49 isolates were composed of 20 isolates of Type 1, 16 of Type 2, 12 of Type 4 and 1 of a new mtDNA type, Type 7, which was closely related to Type 2. On the bases of the results of 120 isolates of the present (49 isolates) and previous (71 isolates) studies, F. pedrosoi was classified into seven mtDNA types and according to the relationship between mtDNA types and geographic origins: in Japan and probably in China, Type 1 isolates; in Zaire and Madagascar, Type 2; in Central and South America, Type 4 and Type 1. These results indicated that the geographical origin of F. pedrosoi isolate could be roughly inferred from its mtDNA type.

Summary 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.

The presence of filamentous bacteria morphologically similar to Nocardia in a fresh stool sample from an AIDS patient with pulmonary nocardiosis is here reported. The material was submitted to our laboratory for a parasitologic examination and was stained by the Kinyoun method, revealing numerous delicate, irregularly stained, branching acid-fast filaments. Nocardia asteroides had been isolated from sputum samples of this patient. The patient was a 32 year-old HIV+ female admitted to our center on June 1997 because of productive cough, right-sided thoracic pain and weight loss. Chest X rays showed the presence of right superior lobe excavated pneumonia. This was the first time we had observed filamentous bacteria similar to Nocardia in a stool sample submitted to parasitologic examination. For similar cases, and when its presence was not detected in other specimens collected from the same patient, intestinal endoscopy and biopsy should be performed for eventual lesions and smear examination repeated with Kinyoun stain and cultures for Nocardia.

Mitochondrial DNA (mtDNA) types based on restriction fragment length polymorphism (RFLP) patterns with HaeIII were investigated in clinical isolates of Sporothrix schenckii in North and South America. In addition to 14 mtDNA types (Types 1-14) so far reported, six new mtDNA types, Types 15-20 were found in this study. Type 3 was divided into two subtypes, Subtype 3A and Subtype 3B based on RFLP with Msp1. Type 14 was also divided into three subtypes, Subtype 14A, Subtype 14B and Subtype 14C based on RFLP with Hha1. Nineteen isolates in the United States consisted of 1 isolate of Type 1, 12 of Type 2, 2 of Type 4, 3 of Type 14 (1 of Subtype 14B and 2 of Subtype 14C) and 1 of Type 15. Twenty nine isolates in Venezuela consisted of 13 of Type 3 (Subtype 3B), 6 of Type 4, 1 of Type 18, 3 of Type 19 and 6 of Type 20. Thirteen isolates in Argentina consisted of 2 of Type 3 (Subtype 3A), 4 of Type 4, 4 of Type 16 and 3 of Type 17. One isolate in Brazil was Type 3 (Subtype 3A). Based on the phylogeny of 20 mtDNA types (Types 1-20) constructed by estimating sequence divergences of mtDNA, mtDNA types were clustered into two groups: Group A (Types 1-3, Type 11 and Types 14-19) and Group B (Types 4-10, Types 12-13 and Type 20). These results suggest that S. schenckii isolates in North and South America mainly belong to Group A.

The international community of mycologists, ISHAM, is faced increasingly with the needs and demands of supporting the development of organizational structures worldwide in the field of medical mycology. This goes beyond the classical objectives of a scientific association, namely organizing international congresses and publishing a journal. In the early part of the next century, this will become a major challenge to ISHAM. Symposia discussing this subject were presented at the 12th Congress of ISHAM held in Adelaide, Australia, in 1994. This paper outlines the reflections and discussions held by members of our Society in 1997.

Medical mycology is of increasing interest to the basic scientist, pathologist, microbiologist and clinician. This interest has been prompted by the rising number of immunosuppressed patients with opportunistic fungal infections, the expanding boundaries of the so-called endemic mycoses, the recognition of several major new endemic mycoses and a variety of other emerging fungal infections, and the development of potent, non-toxic antifungal drugs to treat these infections. The world of mycology is changing dramatically, especially in developing countries which have only limited resources to cope with the impact of the compromised host and the introduction of costly new antifungal drugs. Consequently, there is an urgent need to increase our effectiveness as teachers of medical mycology at all levels and in all regions of the world.

Histoplasmosis is a fungal disease caused by the dimorphous fungus Histoplasma capsulatum (Hc). Cyclophosphamide (Cy) was used as an immunomodulator capable of modifying the course of the disease, as well as of regulating the mechanisms involved in T-lymphocyte mediated immune response. Rats were subjected to intracardiac inoculation of Hc followed by a fractionated treatment with a 100 mg/kg body weight dose of Cy on days +4, +5, +6, +7 and +11 pi. Until day 26 pi, treatment with Cy caused 85% mortality whereas no mortality was observed among animals only inoculated with Hc. On day 14 pi, the group of Hc animals showed a delayed hypersensitivity test (DH) of 26.60 + 13.96 as determined by the swelling of the leg. Conversely, DH was significantly depressed in rats inoculated with Hc and treated with Cy: 3.88 +/- 1.00 (p0.01). Colony forming units count in this group was 2020 CFU/g of spleen, and 24 CFU/g of spleen (p0.01) in controls. A macroscopic study of the organs revealed that the animals in the Hc+Cy group had spleenomegaly and lungs with granuloma and hemorrhagic spots. The controls only presented small lung abscesses. These findings lead to the conclusion that Cy causes a deterioration of cell mediated immune response which results in the manifestation of an acute, fatal experimental mycosis.

The possible adverse effects of chronic, high-dose fluconazole therapy are detailed from analysis of a multicenter, dose-escalating study of the therapy of invasive mycoses. Ninety-three adult patients were studied, 48 of these receivedor = 6 months therapy and 20 receivedor = 1 year. Fifty-eight patients receivedor = 300 mg/day, and 7 receivedor = 600 mg/day. One patient received 1,997 g over 86 months. Twenty-seven percent experienced possible symptomatic side effects, which resulted in 2 patients discontinuing therapy, and 42% had asymptomatic laboratory abnormalities, none of which were progressive. Headache, hair loss and anorexia were the most common symptoms experienced (each by 3% of patients), and eosinophilia and aspartate aminotransferase increases were the most common laboratory findings (12 and 10%, respectively). Fluconazole appears well tolerated and safe in these doses and durations.

A case of cutaneous hyalohyphomycosis, due to Fusarium oxysporum, in a 40 years old man is presented. The patient came from Paraguay where he worked in a tropical rural area. His disease had begun 2 months before his admission as a skin ulcer located in the left leg. Clinical characteristics, diagnosis methods, differential diagnosis with other ulcers of the legs in tropical areas as well as therapeutic measures are discussed in this presentation.

Some epidemiologic, diagnostic and immunologic aspects of 93 (75 males and 18 women) cases of cryptococcosis (CRY) associated with AIDS, hospitalized in the Muñiz Hospital (MH) in Buenos Aires during 1994, were retrospectively studied. The median age (MA) of the patients under study was 28 years (17-49 y); 30 (17-49) y for males and 23 (22-34) y for women. Intravenous drug addiction (34% of patients) and homo/bisexuality in men (17%) were the most frequent risk factors for HIV infection. The MA of these groups were 27.5 (17-41) y and 34 (25-41) y, respectively. Microscopic CSF examination with India ink and the blood cultures (lysis-centrifugation) achieved the diagnosis of CRY in 67 (72%) and 16 (17%) patients, respectively. At diagnosis, the median titers for Cryptococcus neoformans capsular antigen were 1/1,000, 1/100 and 1/1 in serum, CSF and urine, respectively. Sixty five strains of C. neoformans were insolated from clinical samples; all were identified at variety neoformans employing the culture medium proposed by SalkinHurd and the D-proline assimilation test. At diagnosis, CD4+ lymphocytes counts were50/microliter in 46 patients (83.63%), and were between 51 and 200/microliter in 9 (16.4%). The CD4+/CD8+ ratio was1 in all patients. Most patients were born (83%) and lived at diagnosis (96%) in Buenos Aires (Bs As) city, Bs As outskirts towns and cities located in Bs As province.

Summary. The efficacy of two different daily doses of itraconazole (ITRA) and the combination of flucytosine (5-FC) with ITRA in the treatment of an experimental model of cryptococcosis in hamsters was studied. Five groups of 20 animals each were inoculated by the intracardiac route with 105 cells of Cryptococcus neoformans. Treatment started 3 days after the infection, and was administered by gavage for 30 days. ITRA was applied at a daily dose of 25 mg kg-1 or 50 mg kg-1 and the combination of 5-FC and ITRA was given at 75 mg kg-1 day-1 or 50 mg kg-1 day-1 respectively. One group of 20 hamsters received the vehicle and was used as a control group. Treatment evaluation was based on the following parameters: number of surviving animals 60 days after the infection; presence of encapsulated yeasts on microscopic examination of wet preparations of brain, lungs, liver, spleen and kidneys at necropsy; and brain qualitative (massive seeding) and quantitative cultures (determination of colony forming units, CFU). ITRA 50 (50 mg kg day-1) was the most effective treatment according to the studied parameters; 70% of brain cultures became negative and 95% of the treated hamsters survived to the end of the study period. ITRA efficacy was dose dependent. The combination of ITRA with 5-FC was less effective than administering the drugs separately; the reason for this finding is not known. The results obtained in this study should encourage the use of high doses of ITRA in cases of disseminated cryptococcosis in humans. Zusammenfassung. In einem Tiermodell mit Cryptococcose an Hamstern wurde die Wirksamkeit von Itraconazol in zwei unterschiedlichen Dosierungen (25 mg kg-1 d-1) bzw. 50 mg kg-1 d-1) und die der Kombination von Flucytosin (75 mg kg-1 d-1) + Itraconazol (50 mg kg-1 d-1) untersucht. Die Studie wurde an funt Gruppen zu je 20 Tieren durchgefuhrt. die intrakardial mit 105Cryptococcus neoformans-Zellen infiziert worden waren. Eine Gruppe von 20 Hamstern erhielt nur das Losungsmittel und diente als Therapiekontrolle. Die Auswertung der antimykotischen Behandlung stutzte sich auf folgende Parameter: Zahl der uberlebenden Tiere 60 d nach Infektion; mikroskopischer Nachweis bekapselter Hefen in Naspraparaten aus Nekropsiematerial von Gehirn, Lunge, Leber, Milz und Nieren; kultureller Nachweis von Cr. neoformans aus Gehirn-Homogenat qualitativ und quantitativ. Itraconazol 50 mg kg-1 d-1 erwies sich als das wirksamste Behandlungsverfahren: es fuhrte zu 70% Negativitat der Gehirnkulturen und 95%uberlebenden Tieren. Die Itraconazol-Wirksamkeit war dosisabhangig. Die Kombination von Itraconazol mit Flucytosin war weniger wirksam als beide Wirkstoffe fur sich allein, fur diesen Befund gibt es keine Erklarung. Die Ergebnisse ermuntern den Einsatz von Itraconazol in hohen Dosen bei der disseminierten Cryptococcose des Menschen.

The goal of this work was to develop in solid medium a fast method to obtain Paracoccidioides brasiliensis (Pb) with a high yield. Four culture media were assayed: Sabouraud honey-agar, Sabouraud dextrose-agar, tomato -agar-medium (TOM) and a medium based on grape pulp. The most exhuberant growth was observed in medium based on grape pulp. Antigen was prepared in microscale at 6, 10 and 15 days incubation of solid cultures and the crude product concentrated by means of Centriplus tubes (Helena, France). Isolated antigens were subjected to polyacrylamide gel electrophoresis, followed by immunolabelling and detection of the characteristic gp45 antigen employing human and Pb-infected rat sera. Best results were observed after 10 days culture in grape medium. None of the other three media afforded comparable results.

Fueron estudiadas en forma comparativa 6 cepas de P. brasiliensis con el propósito 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) aspiración ósea en 1991 (63265), 4) testículo de cobayo 1984(C24), 5) punción-aspiración ganglionaren 1986 (G) y 6) cepa proveniente de la Escola Paulista de Medicina (339). Se prepararon antigenos citoplasmáticos liofilizados de cada una de ellas, en la concentración final de 100 mg/ml y se realizaron pruebas de inmunodifusión frente a 6 sueros patrones positivos de ratas. En este ensayo todos los antígenos presentaron dos ó tres bandas de precipitación. Para estudiar el poder patógeno se inocularon, en total, 120 ratas Wistar, de ambos sexos de 200 g de peso, por via intracardíaca con suspensiones de la fase levaduriforme del P. brasiliensis, en concentraciones de 3x10(7) y 5x10(7) células/ml de cada cepa. Los animales que no murieron espontáneamente fueron sacrificados a los 14,28,42, 56 y 70 dias post-infección y se evaluaron los siguientes parámetros: A) exámenes macro y microscópicos de pulmones, hígado, bazo y riñones; B) cultivos de un pulmón y C) prueba de inmunodifusión con antígeno homólogo. Se consideró además, el porcentaje de muertes espontáneas por cada cepa. Los resultados de estos estudios fueron los siguientes:No se observó relación entre la patogenicidad y la antigenicidad. La cepa más virulenta correspondió a un aislamiento reciente a partir de una forma juvenil grave y la más antigénica fue una cepa, morfológicamente atípica, que no provocó lesiones macroscópicas ni microscópicas en los órganos de las ratas. A comparative study of antigenicity and patho-genicity for rats of six Paracoccidioides brasiliensis strains was carried out. The antigenic capacity "in vitro" of cytoplasmic extract from each strain was determined by immuno-diffusion 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 3x10(7) and 5x10(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 hystologic 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. The most virulent strain was a recent isolate from an acute disseminated form of the juvenil type. A morphological atypic isolate, which produced a very mild experimental infection with viable P. brasiliensis determined 100% of positive immunodifussion tests. The strain 339 did not produce infection in the rats, and the animals inoculated did not presented antibodies. Nevertheless this strain is useful to prepare antigens for Serologic reactions. No correlation between antigenicity or pathogenicity and the time during which these strains were mantained "in vitro" could be established.

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 HE; 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)

One hundred and seventeen patients suffering systemic mycosis and AIDS were studied during 5 years in the Muñiz Hospital of Buenos Aires City. Seventy four of them presented cryptococcosis, 39 histoplasmosis and 4 both mycoses. The following specimens were studied for the diagnosis: skin and mucous membrane scrapings, bone marrow aspirations, bronchial secretions, biopsies of different organs, cerebral spinal fluid and blood cultures. Sera were also collected for serologic tests. A total of 203 samples from patients with histoplasmosis were studied, 46.3% of them showed H. capsulatum in microscopic examinations or in cultures, skin scraping was the most sensitive diagnostic method (94.7% of positive results), followed by biopsies (80%) and bone marrow cultures (42.1%). Specific antibodies were detected in 45.4% of the patients with histoplasmosis, using 2 different antigens and 3 types of serologic reactions (complement fixation test, immunodiffusion and counterimmunoelectrophoresis). A total of 413 samples from patients with cryptococcosis were examined, 69% of them confirmed the diagnosis. The mycologic study of CSF was the most sensitive method of study, since it registered positive results in 89.5%, followed by blood cultures (61.2%), skin scrapings (42.9%), and urine cultures (41.7%). Polysaccharyde antigens from C. neoformans in organic fluids were detected in almost all the cases. The aim of this study is to compare all the suitable diagnostic methods which can be used in systemic mycosis associated with AIDS in order to find the most rapid way of diagnosis.

Itraconazole is a highly lipophilic triazolic compound, scarcely soluble in acidified polyethylene glycol, and soluble in hydroxypropyl-beta-cyclodextrin. It possesses an excellent digestive adsorption and its peak plasma level after oral administration of 100 mg is 0.16 microgram/ml at 3 or 4 h after drug intake. Half-life of itraconazole ranges between 17 to 21 h and 99.8% binds to plasmatic proteins, especially albumin. Metabolization is mainly done in the liver where inactive metabolites are formed with the exception of hydroxy-itraconazole, which exhibits a discrete antifungal activity. Stabilization of blood levels with repeated drug administration is reached at day 14, showing an increase both in plasma concentrations and in its half-life. Tissue levels of itraconazole are 3- to 20-fold higher than plasmatic concentrations, whereas only negligible concentrations are in CSF and urine. In the skin and particularly nails, itraconazole persists for a long time after discontinuation of therapy. Its mechanism of action is similar to other azolic compounds, inhibiting the alpha-14-demethylase of lanosterol which interferes with the synthesis of ergosterol. This drug behaves as a wide spectrum antifungal agent, acting against most pathogenic fungi with the exception of the Zygomycetes. Daily doses vary, according to indications, from 100 to 400 mg. The efficacy and results obtained in dermatomycosis, candidiasis, paracoccidioidomycosis, keratomycosis, sporotrichosis, chromoblastomycosis, coccidioidomycosis, blastomycosis, cryptococcosis, phaeohyphomycosis and maduromycotic mycetomas are detailed.

A urease negative strain of Cryptococcus neoformans was isolated form a patient with AIDS. The identification of the yeast was confirmed by physiological and pathogenicity tests. Clinically, the disseminated cryptococcal infection in our patient was identical to those reported in other patients with AIDS.

Twenty-seven patients suffering AIDS and disseminated histoplasmosis were included in this study, comprising twenty-three males and four females, from 18 to 46 years of age (mean = 32.9). The most frequent clinical manifestations were fever, weight loss, anaemia, skin lesions, pulmonary micronodules, hepatosplenomegaly and adenomegalies. All of them presented other infectious diseases or neoplasias frequently found in AIDS patients. The diagnosis of histoplasmosis was based upon the finding of Histoplasma capsulatum in microscopic examination or in cultures from the following specimens: skin scrapings, bone marrow aspiration, bronchoalveolar lavage, blood cultures, buccal biopsies and lymph node biopsy. Serologic reactions, searching for antibodies, were positive in 11 cases. Itraconazole by oral route, at a daily dose of 200 mg (24 cases) or 400 mg (3 cases), was administered for 6 months. Those patients who were clinically cured after receiving this scheme of treatment were treated with itraconazole 100 mg day-1 as a suppressive therapy. Twenty-three patients were considered responders, 1 as a non-responder and 3 non-assessable. The average survival time was 7.8 months and eleven cases are still alive. Itraconazole proved to be a useful medication in disseminated histoplasmosis associated with AIDS and it was very well tolerated.

Some epidemiological characteristics of 105 cases of cryptococcosis diagnosed in Argentina, between 1981-1990, were studied. Until 1987, the annual number of cases was 4-8. It has increased since 1988 by AIDS influence and reached 35 cases in 1990. The annual number of cases non associated with AIDS has remained in 3-7 cases. The age median of the all patients, AIDS and non AIDS associated cases was 30, 30 and 45 years old, respectively. The 20-39 years old group was the most affected and the age distribution was different in AIDS and non AIDS population. Masculine predominance was more evident in AIDS than in non AIDS associated cases. The predisposing factor was AIDS in 57 patients, another different factor in 20 and unknown in 28 cases. The indirect estimation of the percentage of AIDS cases with cryptococcosis wasor = 6.19% (57 cases in 920 HIV+) during 1981-1990 this period. Cryptococcus neoformans variety neoformans was isolated from 101 cases and the gattii variety (serotype B) from only 4. The obtained data are similar to those observed in Europe and United States.

Eighty-eight immunocompetent patients with deep mycoses from eight countries were evaluated with the same protocol for efficacy of fluconazole monotherapy. Entry doses were raised from 100 to 400 mg as safety was shown in initial cohorts, and dosages up to 2,400 mg daily and durations up to 44 months were studied. Results were very similar in different countries. Twenty-seven of 28 evaluable patients with paracoccidioidomycosis, 13 of 19 with sporotrichosis, 14 of 16 with coccidioidomycosis, and eight of eight with histoplasmosis demonstrated objective responses to therapy, as did one patient each with zygomycosis and alternariosis. For these patients, relapses have been unusual thus far. In contrast, one patient with chromoblastomycosis responded but relapsed, and six did not respond; one patient with mycetoma responded but relapsed, and two did not respond. The drug was well tolerated by patients, including six who received intravenous therapy. In vitro susceptibility tests suggested that clinical response was correlated with susceptibility but that resistance did not preclude clinical response. Fluconazole therapy appears efficacious for several deep mycoses; dosages of greater than 200 mg daily may be needed for some diseases. The further evaluation of fluconazole for these entities is warranted.

An E.L.I.S.A. test for antibody detection, with an exo-antigen of Coccidioides immitis was standardized in 67 humans sera diluted in 1/1000, 1/2000, 1/4000 and 1/8000. Eighteen sera from mycologically proved cases of coccidioidomycosis were studied: 5 were negative and 13 were positive in some dilutions. 3/26 sera of healthy persons who presented positive skin tests with coccidioidin were positive and the other 23 sera did not have positive reactions. None of the 15 sera of healthy human exhibited positive E.L.I.S.A. Serum samples of 8 patients suffering other deep mycosis were studied, 4 of them presented cross-reactions in E.L.I.S.A. tests. E.L.I.S.A. test seems to be a useful serologic technique for antibody detection in anticomplementary serum samples or when a low concentration of antibodies should be detected. As it is very sensitive, cross-reactions with other mycoses are frequent, thus the use other more specific serologic technique together E.L.I.S.A. is recommended.

A new, nontoxic material extracted from Aspergillus fumigatus, is described with complement-depletion capacity both in vivo and in vitro. Extracts of the mycelium of A. fumigatus were found to haemolyse sheep red blood cells, to be lethal for mice and to convert C3 into its faster electrophoretic form when incubated with normal human serum. Conversion did not take place when purified C3 was used, suggesting that serum factors were required in the reaction. Adsorption of the extracts with activated carbon or heating at 100 °C for 30 min yielded detoxified materials devoid of both toxic and haemolytic properties but still capable of converting serum C3. In guinea pigs, administration of detoxified extracts was followed by complement depletion lasting not less than 48 h. Heating (100 °C for 30 min) extracts which had been previously detoxified by carbon adsorption resulted in a partial loss (40%) of their complement-inhibitory capacity indicating that part of the anticomplementarity of AFE was due to a heat-stable substance.

The author presents the drugs that are available for the treatment of opportunistic mycoses: amphotericin B, nystatin, 5-fluorocytosine, miconazole and the newest imidazole derivative econazole. He presents his experience with econazole in 4 cases with deep mycoses. He speaks of the mode of application and the therapeutical limits of these products as well as of the favorable factors and the prophylactic measures to be taken.

Some of the biological properties of saline extracts of the mycelia of several species of the Aspergillus genus, namely, A. fumigatus, A. flavus, A. niger, A. nidulans, A. parasitucus and A. glaucus, were studied. Only the extract prepared from A. fumigatus was found to be hemolytic for sheep red blood cells. In contrast, all the extracts with the only exception of that of A. glaucus, had cytotoxic effects on Vero cells. Both, the hemolytic and cytotoxic constituents of the extracts were removable by adsorption with activated carbon. Heating of the extracts at 100 degrees C for 30 minutes also resulted in detoxification. In vivo studies, performed only with detoxified extracts of A. fumigatus, showed these were capable of depleting complement levels in guinea pigs. Complement inactivation was also found to occur in vitro and was cuased by all the extracts tested. Also triggered by the extracts was the conversion of serum C3 but not of purified C3, indicating that other serum factors are essential in the process. Despite the similarity in this respect with cobra venom factor, differences in activity after heating-negative in cobra venom factor-indicate that the complement inactivating substance/s present in the Aspergillus extracts differ from those of the snake product.

The results of skin tests with two antigens of the yeast phase of Histoplasma capsulatum are presented. Both antigens were able to fix complement and form precipitating bands in the presence of sera from patients with active histoplasmosis. Their sensitivity was lower than the metabolic antigen usually employed in serology. Results of skin tests obtained with both yeast phase antigens and a standard histoplasmin coincided in animals infected with H. capsulatum. Only one cross reaction was observed in animals inoculated with Paracoccidioides brasiliensis. Skin tests in humans were conducted on patients with and without mycotic disease. The percentages of positive reactions in patients with histoplasmosis were not significantly different between control histoplasmin and whole cell extracts. In the patients with non mycotic diseases the frequency of positive tests varied between 28.8% and 32%, which agrees with previous statistical data for the general population of the area. Equally the positive percentage of 55% and 68% in patients with paracoccidioidomycosis coincided with the results of previous studies. The histological patterns of these skin tests showed that they were produced by cell mediated hypersensitivity. The sensitivity of a whole yeast cell extract was similar to histoplasmin L48, but its preparation was quicker and easier to perform and it had no foreign substances from the culture medium, so we think that it would be easier to standardize it chemically.

Results are presented of treatment with miconazole, orally and intravenously, in patients with paracoccidioidomycosis. Twenty-eight male patients aged from 34 to 66 years and exhibiting various clinical forms of the disease were studied. Twenty-five came from endemic areas in north east Argentina (Chaco, Formosa, Misiones, Corrientes and northern Santa Fe) and the remaining three from Paraguay. Twenty patients were engaged in agricultural work or at woodmills. single or multiple lesions were observed in 24 cases. Thirteen were suffering from infection of the larynx and in two of them a tracheotomy was necessary. Twenty-three showed pulmonary lesions on X-rays. Twelve had ganglionic lesions, eight had cutaneous lesions and one patient had osteoarthritis of the knee. One patient had hepatomegaly which was unrelated to chronic alcoholism. Fourteen patients had received previous treatments such as sulphonamides and amphotericin B (7 cases); sulphonamides (3), sulphonamides and the combination sulfamethoxazole + trimethoprim (3), and one patient had received all three medications. All patients had relapsed before starting miconazole therapy. Diagnosis was established by the presence of P. brasiliensis in all cases, recovered either from cutaneous or mucosal biopsy samples or from the sputum. Complement fixation tests were positive in all patients at the onset of the treatment and the immunodiffusion reactions showed precipitation bands in 27/28 patients. Skin tests with P. brasiliensis antigens proved to be positive in 18 cases and negative in 10. The erythrocyte sedimentation rate was markedly accelerated in 22 patients (greater than 20 mm in the first hour).(ABSTRACT TRUNCATED AT 250 WORDS)

The extent of complement depletion by Aspergillus fumigatus extracts (AFE) was smaller in hypogammaglobulinemic than in normal human sera, suggesting that γ-globulins play a role in this phenomenon. The titers of Cl, C4 and C2 were reduced after reaction with AFE indicating that complement activation followed the classic pathway. The alternate pathway was also involved since AFE caused conversion of both C3 and factor B in the sera of Clr- and C2-deficient patients and in normal human serum in the presence of EGTA. Similar conversions were observed with hypogammaglobulinemic sera. Heating the AFE at 100°C for 30 min abolished their ability to activate the classic but not the alternate pathway. Hydrazine-sensitive factors were required by AFE to activate the C3 bypass.

The sera from normal subjects gave negative results with the following antigens used in the complement-fixation tests: 1) polysaccharide prepared according toFava Netto's technic; 2) a filtrate of shaked cultures followingAjello et al.'s technic; 3) an aqueous extract of mechanically disrupted yeast cells ofP. brasiliensis.

SummaryStudies have been made on a case of septicaemia produced by a yeast-like fungus classified as Torulopsis glabrata (Anderson). The patient was a girl suffering from “myasthenia gravis”. She presented a mediastinal empyema and pleural effusion following a thymectomy. Antibiotics were administered. Serology revealed a high titre of circulating antibodies to T. glabrata and cross reactions with Candida albicans during the acute phase of the mycosis. The patient was successfully treated with amphotericin B, the titre of circulating antibodies decreasing with improvement of the mycotic condition.

Male guinea-pigs were inoculated by the testicular route with a suspension of chlamydo-arthrospores of the filamentous phase ofC. immitis. The following histopathological changes were observed: voluminous pyocytic foci as well as granulomatous mononuclear reaction was initially observed and granuloma with multinucleated giant cells were seen 20 days after the inoculation. The following changes of the microscopic aspect of the parasite were correlatively registered: the so-called primary infection type of sporangia appear in great number three days after the inoculation. This type of sporangium is characterized by its great diameter up to 98 µ, peripheral endospore formation leaving a large central vacuole which frequently contain residual protoplasm. The endospores are thin walled, polyhedral and get free through an ostiole. Sporangia completely filled with globose endospores (the cystic type of sporangia) with radiate acidophilic formations on the peridial wall were observed in the testicles of the guinea-pigs killed 6 days after the inoculation. Reduction in number and in the size of the parasite were seen after the 20th. day of the inoculation. Radiate formation of the cell wall of the parasite appeared simultaneously with precipitin antibodies and would be the expression of antigen-antibody reaction.

Ketoconazole was administered orally in daily doses of 200 mg to 70 patients with onychomycosis. Complete recovery was attained by 48 patients, improvement of > 50% by 10 patients, and for three patients therapy failed. For the remaining nine patients the results could not be evaluated. The average duration of treatment was 7.5 months for disease due to Trichophyton species and 6.5 months for disease due to Candida species. There were no adverse effects or signs of toxicity attributable to the administration of ketoconazole. It is concluded that ketoconazole is a positive development in the effort to control the difficult problems presented by onychomycosis.

329 adult patients with tinea pedis interdigitalis, which clinical diagnosis was confirmed by positive direct microscopic examinations and cultures of organisms from the lesions, were enrolled in this open and multicentric study. Informed consent was requested and obtained in each case. The bifonazole, 1% gel, tested was applied topically once daily for 3 weeks. Weekly clinical controls were performed during the treatment period and, at least, two clinical mycological posttreatment assessments were carried out. The evaluation criteria were based on the clinical and mycological findings at the end of the second posttreatment week. According to these criteria, the results obtained were: very good (clinical cure, negative mycology) 276 (84.0%); good (clinical improvement, negative mycology) 32 (9.5%); moderate (clinical improvement, positive mycology) 20 (6.0%), and failure (no clinical and mycological changes) 1 (0.5%). Tolerance was excellent, side effects were not observed in any case.

With the objectives of determining the effectiveness of bifonazole 1% powder in the prophylactic treatment of tinea pedis interdigitalis and its local tolerance, 200 patients who achieved clinical and mycological cure in another previous trial with bifonazole 1% gel, and who were also reliable in the compliance of the treatment, were included in this study. Informed consent was requested and obtained in each case. This was a multicenter, double-blind, controlled and comparative study of one group (100 patients) treated with bifonazole verum against another group (100 patients) treated with bifonazole placebo. The allocation to each group was performed by randomization. Bifonazole was applied once daily in the morning on both feet, after showering, and inside shoes; patients applied it again when changing shoes or socks during the day. Treatment lasted up to 6 months. During the treatment, patients were clinically and mycologically assessed once a month and, if it was necessary, when signs and/or symptoms of infection reappeared. The evaluation criteria were based on the reinfection, clinically and mycologically demonstrable, during the treatment. 29 patients abandoned the treatment. Reinfection was observed in 57 out of the 171 patients who finished the treatment, 47 reinfections occurred in the placebo group and the remaining 10 in the verum group. Tolerance was excellent, no side effects were observed in any patient.

A cellular antigen from Paracoccidioides brasiliensis, obtained from the supernate of the disrupted yeast phase, was studied. It was prepared by a method similar to that for obtaining the spherulin from Coccidioides immitis. This antigen is composed of 55 mug/mg of polysaccharides, determined by the phenol method, and 7-8 mug/mg of protein, determined by Folin-Ciocalteu's technique. In vitro tests of antigenicity were done by immunodiffusion and complement fixation. The optimum dilution of the antigen was 0-5 mg/ml determined by microtiter complement fixation. The in vivo standardization of the cellular paracoccidioidin was performed on three groups of patients. The first was composed of 56 patients with mycologically proved paracoccidioidomycosis. The disease was active in 32 and clinically healed in 24 of this group. The second was composed of 15 cases of histoplasmosis with strong histoplasmin skin tests. The third group were 70 patients with no mycotic pulmonary diseases who had come from different zones in Argentina. Each individual was injected with 0-1 ml cellular paracoccidioidin (3 mg/ml), 0-1 ml of Fava Netto's antigen diluted 1/10, and 0-1 ml histoplasmin L48, diluted 1/100. The cellular paracoccidioidin skin test was positive in 44/56 (78.5%) patients with paracoccidioidomycosis. The Fava Netto's antigen gave 42 (75%) positive results and the histoplasmin skin tests gave 28 (50%) in the same group. Twenty-eight (50%) presented positive histoplasmin skin tests. The paracoccidioidin skin test was positive in 2 of the histoplasmosis group (13-3%) and the Fava Netto's antigen gave 3 positive results in the same group (20%). The histoplasmin skin test was positive in 32% of the individuals with no mycotic disease. Ten per cent of these exhibited positive reaction with both P. brasiliensis antigens. Of those paracoccidioidomycosis cases (Group 1) who had negative skin tests with 3 mg/ml of the cellular paracoccidioidin 7/10 gave positive skin tests when tested at a concentration of 8 mg/ml. Characteristic changes of cell medicated hypersensitivity were observed in the histopathological studies of the cellular paracoccidioidin skin test.

The ultrastructural changes produced by ketoconazole in the yeast-phase of H. capsulatum and P. brasiliensis were studied by means of scanning and transmission electron microscopy. The observed alterations on both fungi were very similar to those induced by the same drug on the ultrastructure of C. albicans. These alterations include surface changes, abnormal membrane proliferation, fatty degeneration of the cytoplasm and lysis of the subcellular organelles. P. brasiliensis seems to be more sensitive to ketoconazole than H. capsulatum, since the necrosis of most of the cells was obtained in the former at a concentration of 0.1 microgram/ml and in the latter at 1 microgram/ml.

A comparative study of the "in vitro" susceptibility of 24 Cryptococcus strains to 5 antifungal drugs (amphotericin B, 5 fluorocytosine, miconazole, itraconazole and ketoconazole), was carried out. These strains were grouped according to species, varieties and isolation's origins. The minimum inhibitory concentration (M.I.C.) was determined by the agar dilution technique in yeast nitrogen base agar with dextrose. The mean geometrical of the M.I.C. values of each group was compared with the others. The results obtained were homogeneous with the only exception of the "non neoformans" strains, in which, higher M.I.C. to 5 fluorocytosine values were detected.