Combined anti-fungal therapy and surgical resection as treatment of pulmonary zygomycosis in allogeneic bone marrow transplantation.

Opportunistic fungal infection is a rare but severe complication in allogeneic bone marrow transplant (BMT) recipients. We report a 49-year-old patient who developed pneumonitis after BMT, due to a Mucorales fungus (class Zygomycetes), Absidia corymbifera. Infections due to mucormycosis are likely to become increasingly recognized even though the occurrence after BMT has only been described sporadically. We postulate that the patient was contaminated before BMT despite no intensive drug treatment or other iatrogenic features, related to his poor living conditions and developed the infection during aplasia. He immediately received i.v. liposomal amphotericin B (AmBisome) and GM-CSF. Because there was no response, the infected area and necrotic tissue were resected. Despite initial clinical and biological improvement and the absence of Mucor on mycological examination post-surgery, the patient died 3 weeks later from bilateral pulmonary infection and multiorgan failure.

Purification of a specific antigen of Trypanosoma cruzi from culture forms.

A monospecific rabbit antiserum was prepared against a specific component of Tryanosoma cruzi epimastigote culture forms (designated antigen 5). Rabbits were immunized intradermally in 30-50 sites with the zone of agarose gel in which the component 5:anti-5 complex was visualized after two-dimensional immunoelectrophoresis. With this appropriate antiserum and the total epimastigote soluble extracts, the specific component of T. cruzi was purified by immunoadsorption. Purity and specificity controls of thefinal product were carried out by prolonged immunization of rabbits, and then by immunoelectrophoretic and two-dimensional electrophoretic analyses.

A case of chromomycosis treated by a combination of cryotherapy, shaving, oral 5-fluorocytosine, and oral amphotericin B.

A case of chromomycosis from Comoro Islands was first treated without success with high doses of oral amphotericin B (3 g per day). Treatment with itraconazole (400 mg per day) was also unsuccessful. Then, in vitro tests were done to study the susceptibility of this Fonsecaea pedrosoi strain to antifungal drugs. It was resistant to itraconazole, sensitive to 5-fluorocytosine, and the combination of 5-fluorocytosine with amphotericin B was synergistic. The patient was then treated with this last combination of drugs, which seemed to be effective. The patient stopped this treatment after six months, and relapse occurred two years later. The best therapeutic strategy in cases of chromomycosis seems to be a combination of two drugs chosen according to the results of prior antifungal susceptibility testing.

Antigenic make-up of Trypanosoma cruzi culture forms: identification of a specific component.

A comparison of Trypanosoma cruzi water soluble antigens with those of stercorarian and salivarian trypanosomes, and Leishmania using immunoprecipitation in gels and immunoelectrophoresis, with the aid of hyperimmune rabbit serum and heterologous adsorptions showed the following. 1) There is a high complexity of soluble antigens of T. cruzi and T. rangeli. 2) At the intraspecific level our results demonstrated the antigenic stability of T. cruzi when maintained in vitro, and that there was quantitative antigenic consistency of the culture forms of different strains of T. cruzi from diverse geographic and parasite sources. At the interspecific level, the antigenic relationships between T. cruzi and the other Trypanosomatidae were established, as follows: 6/10ths of the antigens are shared by stercorarian species (T. dionisii, T. rangeli); 4/10ths by a salivarian trypanosome (T. brucei); and 3/10ths by Leishmania (L. donovani, L. mexicana). 3) Among the 4/10ths of antigenic components specific to T. cruzi, one component was characterized by its antigenicity and immunogenicity in natural and experimental infections, and in immunization experiments; this component was specific to T. cruzi when compared to the other Trypanosomatidae antigens.

[Detection of an antigen related to systemic Candida infection using a monoclonal antibody conjugated to colloidal gold]. / Détection d'un antigène témoin d'infection systémique à Candida à l'aide d'un anticorps monoclonal couplé à l' or colloïdal.

Previously we showed it was possible to detect antigenemia associated with systemic candidiasis using an anti-C. albicans monoclonal antibody conjugated to colloidal gold. The technique being used, known as Immuno-Gold-Silver staining (IGSS), is applied to serum dots on cellulose nitrate. It is very simple in practice, the results of the reaction being visible with the naked eye. The diagnostic value of IGSS has been compared, on the one hand, with that of the anti-Candida antibody detection by co-counterimmunoelectrophoresis and indirect immunofluorescence, on the other hand with that of the antigen detection using the Cand-Tec commercial test. The specificity and sensitivity of these methods have been established in relation to sera of 79 subjects shared out into 4 groups: sound-subjects, patients having developed systemic candidiasis following surgery, leukemic patients apparently uninfected with Candida and leukemic patients suffering from systemic candidiasis. The IGSS which is slightly less specific than the Cand Tec makes it possible to diagnose a much greater number of infections. Selected bioclinical observations show that there exists complementarity between the detection tests of antibodies and those of antigens and that it is possible to attribute a prognosis value to antigenemia detected with the IGSS dot method.

[Fungal infections: involved species and minimal inhibitory concentrations of common antifungal agents]. / Infections à levures: espèces concernées et concentrations minimales inhibitrices des antifongiques usuels.

Fungal cultures of cutaneous or ungual origin (477 cultures), nasopharyngeal and urogenital origin (2,000 cultures), and blood, internal organs, surgical incisions, and catheters (300 cultures) were obtained in this study. Analysis of the data yielded the following information: Candida albicans and C. tropicalis, frequent causes of superficial or systemic mycoses, were very sensitive to flucytosine and amphotericin B in a liquid medium, but less sensitive to the imidazole derivatives; C. parapsilosis, a cause of superficial and systemic mycoses, was remarkably sensitive to all four antifungal agents tested; Torulopsis glabrata and C. krusei are of greatest concern in a hospital setting since systemic or visceral infections are minimally sensitive to antifungal agents; C. pseudotropicalis, C. guilliermondii, and C. zeylanoides are less pathogenic and sensitive to antifungal agents varies, depending on the strain. Specific measures are presented for management of fungal infections in a hospital setting. The role of antifungal susceptibility testing in everyday practice is also evaluated.

[Application of a co-electrosyneresis reaction to the diagnosis and serological surveillance of candidiasis in a hospital milieu]. / Applications d'une réaction de co-électrosynérèse au diagnostic et à la surveillance sérologique des candidoses en milieu hospitalier.

We have previously described a co-immunofiltration reaction which identifies a specific precipitating system (SPCS) in sera from patients presenting systemic candidiasis. The SPCS is characterized by coalescence with an experimental serum directed against the germinative tubes of Candida albicans. The present study concerns our experience of the use of co-immunofiltration in routine hospital practice. Complete observations involving clinical, mycological and serologic data were selected in order to illustrate various possible developmental trends for SPCS during candidiasis. The SPCS usually develops early infections due to the yeast species most frequently implicated in hospital pathologies; an increase in its intensity reflects a developing infection or the start of therapy. The SPCS disappears slowly and gradually when infectious development is favorable, but its sudden disappearance represents an unfavorable prognosis correlated with detection of circulating antigens. Circulating antigens are also seen in severe cases of candidiasis in which the SPCS is not present. Indeed there is a certain complementarity between these two observations.

[Occupational extrinsic allergic alveolitis due to Aspergillus oryzae]. / Alvéolite allergique extrinsèque professionnelle à Aspergillus oryzae.

We report a case of extrinsic allergic alveolitis, provoked by Deterzyme, a product used in dermatology for the cleaning of cutaneous sores, and containing fractions of protolytic enzymes and amylases of Aspergillus oryzae. The diagnosis was based on the positive precipitins to an extract of antigen of the product as well as positive bronchial provocation tests (semi-delayed) and the disappearance of the symptomatology without any sequelae following cessation of exposure to the risk.

Expression of an epitope by surface glycoproteins of Candida albicans. Variability among species, strains and yeast cells of the genus Candida.

Expression of an antigenic epitope reactive with an IgM monoclonal antibody was studied in 253 Candida isolates by direct agglutination. Isolates of Candida albicans reacted significantly more often with the antibody than did isolates of other species. The agglutination scores for C. albicans isolates from sources associated with possible deep-seated Candida infection were significantly higher than for those from other sources. However, there was considerable overlap of scores between these two groups so that the functional significance of the association is uncertain. There were no associations between agglutination score and C. albicans biotype: multiple isolates from individual patients gave similar agglutination scores. Expression of the epitope within colonies of cloned strains was studied by autoradiography of colony blots on nitrocellulose and by immunoperoxidase staining of colonies in situ. It was found that the epitope was expressed variably by portions of colonies of both agglutination-positive and agglutination-negative strains. Indirect immunofluorescence revealed that expression of the epitope at the surface of C. albicans germ tubes and their associated buds was variable from cell to cell. It was not directly related to morphology or stage of growth of the cells.

Retrospective evaluation of two latex agglutination tests for detection of circulating antigens during invasive candidosis.

Two latex agglutination tests for the detection of Candida antigens, Pastorex Candida (Sanofi Diagnostics Pasteur, Marnes-la-Coquette, France) and Cand-Tec (Ramco Laboratories, Inc., Houston, Tex.), were applied to 79 serum samples from 19 patients who were retrospectively selected on the basis of mycological and clinical evidence of C. albicans infection and the availability of serial serum samples taken near the date of a positive culture. The specificity in 60 control individuals was 100% for Pastorex and 98.3% for Cand-Tec. The tests scored positive for 10 (52.6%) and 9 (47.4%) patients, respectively. Pastorex detected antigen in only 3 of 12 patients (25%) with positive antibody detection tests, but was positive for all 7 patients (100%) who produced no or a low antibody response, suggesting that the test performs better in the absence of antibodies. However, the sensitivity of Pastorex also increased with the number of samples available per patient, which was lower for high-antibody-responder patients (2.8 versus 5.7). If the patients who provided only one or two serum samples were eliminated, the sensitivity of Pastorex rose to 76.9%. For the Cand-Tec, the sensitivity was not related to the presence of antibodies, nor was it related to the number of samples per patient. The observed antigenemia was transient with both Pastorex and Cand-Tec. Only 12.5% of the positive reactions occurred on the same serum sample, confirming that the two tests react with different antigens. A positive antigen test preceded other diagnostic indications for 6 of 10 Pastorex-positive patients and 5 of 9 Cand-Tec-positive patients.

[Comparative immunoelectrophoretic analysis of culture and bloodstream forms of "trypanosoma" (Trypanozoon) brucei gambiense". Demonstration of stage and antigenic type specific antigens (author's transl)]. / Analyse immunoélectrophorétique comparative des formes de culture et des formes sanguicoles de Trypanosoma (Trypanozoon) brucei gambiense. Caractérisation d'antigènes spécifiques de stade et de type antigénique

The comparative immunoelectrophoretic analysis of the antigenic structures of culture and bloodstream forms with cross absorption tests confirm the existence among Trypanosoma gambiense antigens of: --numerous antigenic components common to culture and bloodstream forms;--several antigenic components specific of either culture forms or bloodstream forms. Among the latter, two components are shown to be specific of the antigenic type and appear to support the immunochemical specificity of each variant.

Identification and purification of the specific antigen of Paracoccidioides brasiliensis responsible for immunoelectrophoretic band E.

A new purified antigen (E2) of Paracoccidioides brasiliensis mycelial growth phase was isolated by immunoadsorption from a crude metabolic soluble extract of the fungus. The antiserum prepared in a rabbit by inoculation of E2 antigen developed only one immunodiffusion line with the crude metabolic extract. Findings on immunological analysis showed that E2 antigen is the antigenic component of immunoelectrophoretic band E. The isolated antigens did not possess detectable alkaline phosphatase activity. It reacted in immunodiffusion tests with all the sera (14/14) from P. brasiliensis infected patients containing precipitating antibodies.

Peanut agglutinin affinity chromatography of Trypanosoma cruzi glycoproteins.

Glycoproteins from Trypanosoma cruzi epimastigotes have been extracted by diiodosalycilic acid and lithium salts, and phenol-water biphasic partition. Peanut agglutinin has been used in a one step preparative method for fractionating the total extract in order to separate the so-called galactose-terminal glycoproteins. The different fractions have been studied by SDS electrophoresis, ultracentrifugation and immunoelectrophoresis techniques. The experimental immunogenicity, antigenicity and specificity of the PNA affinity fractions has been evaluated.

Diagnosis of systemic candidiasis: application of co-counterimmunoelectrophoresis.

The accuracy of co-counterimmunoelectrophoresis was compared with mycological and clinical data for different groups of inpatients. The specificity and sensitivity of this method ranged from 100 to 69%. The co-specific precipitin line was observed during systemic infections caused by most opportunistic Candida species. The variations in the line in successive serum samples provided a simple means of serological surveillance. The test also detected seroconversion. The gradual disappearance of the line reflected a favourable prognosis, whereas its abrupt disappearance indicated unfavourable prognosis and correlated with the appearance of detectable circulating antigens.

New enzyme immunoassays for sensitive detection of circulating Candida albicans mannan and antimannan antibodies: useful combined test for diagnosis of systemic candidiasis.

Two standardized enzyme immunoassays for the serological diagnosis of candidiasis were developed. The first one detects antimannan antibodies, while the second one detects mannan with a sensitivity of 0.1 ng/ml. These tests were applied to 162 serum samples retrospectively selected from 43 patients with mycologically and clinically proven candidiasis caused by Candida albicans. Forty-three serum samples were positive for mannan, and 63 had significant antibody levels. Strikingly, only five serum samples were simultaneously positive by both tests. When the results were analyzed per patient, 36 (84%) presented at least one serum positive by one test. For 30 of them, positivity by one test was always associated with negative results by the other test for any of the tested sera. For six patients whose sera were positive for either an antigen or an antibody response, a balance between positivity by each test was evidenced by kinetic analysis of sera drawn during the time course of the infection. Controls consisted of 98 serum samples from healthy individuals, 93 serum samples from patients hospitalized in intensive care units, and 39 serum samples from patients with deep mycoses. The sensitivities and specificities were 40 and 98% and 53 and 94% for mannanemia or antibody detection, respectively. These values reached 80 and 93%, respectively, when the results of both tests were combined. These observations, which clearly demonstrate a disparity between circulation of a given mannan catabolite and antimannan antibody response, suggest that use of both enzyme immunoassays may be useful for the routine diagnosis of candidiasis.

Antigenic analysis of a variant specific component of Trypanosoma Brucei Brucei I -- Immunochemical purification by affinity chromatography.

An antigenic component specific to the bloodstream form of serotype AnTat-1 of Trypanosoma brucei brucei was purified by immunoadsorption technique. Subsequently, a monospecific rabbit antiserum was prepared. Purity and specificity controls of the final product were carried out by immunoelectrophoretic analysis.

Scedosporium (Monosporium) apiospermum: multiple brain abscesses.

A 32-year-old female with alcoholic cirrhosis attempted to drown herself in a pond. She remained in a coma for 57 days. After a stormy course she developed multiple brain abscesses due to Scedosporium (Monosporium) apiospermum. Despite intravenous and intrathecal amphotericin B (total dose 870 mg) the patient died 71 days after the near drowning.

Kinetic study of antifungal activity of amphotericin B, 5-fluorocytosine and ketoconazole against clinical yeast isolates using liquid-phase turbidimetry.

The fungal growth of clinical yeast isolates and of VW32 clone of Candida albicans were measured in vitro using a liquid-phase turbidimetric system (Bioscreen from Labsystems, France) in defined conditions. Cultures were performed in Shadomy's liquid medium and the fungal growth automatically evaluated every 10 minutes for 24 hours using various concentrations of drugs. The system made it possible to test 200 culture samples in one experiment. Yeast sensitivity to drugs was also measured by using our routine semi-automatic turbidimetric system. We observed that kinetic patterns of activity of each antifungal agent were typical. The in vitro tests showed that of 927 clinical yeast isolates 99.2% were sensitive to amphotericin B, 94.4% to 5-fluorocytosine and 69.7% to ketoconazole.