[Molds in the masonry].
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Biomedical subjects
Publications and source records attributed to F Staib.
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A serine proteinase (Alp) from the culture supernate of a clinical isolate of Aspergillus fumigatus was purified to virtual homogeneity at a yield of 41%. The procedure involved affinity chromatography on agarose-epsilon-amino-caproyl-D-tryptophan methyl ester. Alp had an estimated mol. wt of 32 Kda and the pI was determined at pH 7.9. The enzyme was fully inhibited by phenylmethyl sulphonyl fluoride, chymostatin and alpha-1-proteinase inhibitor, and it was largely inhibited by alpha-1-anti-chymotrypsin. Partial inhibition was observed with tosyl-phenylalanine chloromethyl ketone, but tosyl-lysine chloromethyl ketone was ineffective. Thus, Alp may be identical with the major chymotryptic activity of A. fumigatus, which has already been described. The N-terminal sequence of 25 amino acids revealed an 88% homology of Alp with the subtilisin-related proteinase of A. oryzae. Alp acted on casein over a broad range from pH 5.5 to 11.5 and also acts to a lesser extent on haemoglobin and serum albumin. The enzyme degraded elastin and a synthetic elastase substrate; hence, it may be identical with the previously described elastinolytic activity of the fungus. At pH 7.3 and a concentration of 1 microgram/ml, Alp was not toxic for Vero cells, but it efficiently detached such cells from a plastic surface. Specific antibodies against Alp were detected by enzyme immunoassay in the sera of patients and Alp-antigen was demonstrated by immunofluorescence in mycotic human lung. In addition, a second proteinase (Exalp) with extremely alkaline activity, and an aspartic proteinase of A. fumigatus are described.
In an 84-year-old man cryptococcosis with a pronounced osteomyelitic course (involvement of a rib, two thoracic vertebrae, the sacrum and a femur) was diagnosed histologically post mortem, thus directing attention to cryptococcosis in advanced age. The presumptive chronic course of the Cryptococcus neoformans (Cr. n.) infection in this patient was ascribed to chronic kidney failure, diabetes mellitus and chronic obstructive lung disease. Special reference is made to the mycological and radiological diagnosis of cryptococcosis of the bones as well as to its pathogenesis, therapy and epidemiology.
A 40-year old homosexual AIDS patient recovering from a Pneumocystis carinii pneumonia developed a Cryptococcus neoformans infection with involvement of the central nervous system (CNS) which could be treated successfully with amphotericin B and flucytosine. After a symptom-free interval of 4 1/2 months, a new acute fatal disease of the CNS did not reveal a cryptococcosis relapse but a necrotizing Toxoplasma encephalitis, a cytomegalovirus infection and striking cultural findings of Staphylococcus aureus in all organs examined. Neither by culture nor by histology Cr. neoformans could be detected in the CNS or in the other organs examined. The temporal course of the Cr. neoformans infection and its specific diagnosis are commented. It is demonstrated that (during or after successful therapy of Pneumocystis carinii pneumonia) a specific cultural examination of specimens from the respiratory tract for Cr. neoformans is needed, in order to recognize a Cr. neoformans infection in its primary stage, i.e. before hematogenous dissemination of Cr. neoformans leading to the secondary stage of the infection.
The clinical course and response to therapy of seven patients with cryptococcosis and AIDS were reviewed. One patient was still in the primary stage of cryptococcosis in AIDS, i.e. the stage that is characterized by the sole cultural detection of Cryptococcus neoformans in the respiratory tract. The other six patients were in the secondary stage, where C. neoformans can be detected from the cerebrospinal fluid (CSF), blood, urine, faeces and other body sites. The main presenting features (headache, fever, nausea) were due to central nervous system involvement, although meningism and mental changes were rarely present, and CSF changes were very subtle. Treatment with amphotericin B and flucytosine was very effective, there being no more growth of fungi in cultures in most cases. Adverse reactions to the drugs used occurred frequently and consisted mainly of anaemia, hepatosis and fever. Diagnosis in the primary stage of cryptococcosis may improve the prognosis.
Because of the known pathogenicity of Cryptococcus neoformans and of aspergilli depending on defined but different immunodeficiencies of the host, the evaluation of their simultaneous cultural detection in specimens of the respiratory tract of AIDS patients is of epidemiological, diagnostic, pathogenetic and therapeutic interest. In 10 out of 15 AIDS patients the following species of the genus Aspergillus could be isolated either once or repeatedly during the course of Cr. neoformans infections in specimens from the respiratory tract: A. fumigatus, A. flavus, A. nidulans, A. niger and A. glaucus. From the 10 different case reports the conclusion can be drawn that the cultural findings of aspergilli in specimens from the respiratory tract during the course of cryptococcosis were of no pathogenic significance. Rather, they should be interpreted as inhaled and not yet germinated conidia still capable of growth lacking primary pathogenicity for the AIDS patient. The significance of serodiagnosis for the evaluation of the simultaneous occurrence of Cr. neoformans and Aspergillus spp. is demonstrated and discussed.
For the cultural control of Cryptococcus neoformans (Cr.n.), among the routinely examined standard specimens like CSF, sputum, blood, etc., urine earns special attention. The combination of membrane filtration technique (MFT) and Staib agar for the detection of Cr.n. from body fluids as described by Staib in 1963 was used for the cultural isolation of Cr.n. from urine of AIDS patients. In 3 examplary cases the diagnostic significance of this method could be demonstrated: The brown colour effect (BCE) of Cr.n. of a single CFU, as well as in colonies growing with a high density, was produced on average within 3-5 d/26 degrees C. The method was found to be useful for the evaluation of antimycotic therapy. One example of the survival of a few CFUs of Cr.n. under treatment with fluconazole as compared to the efficacy of therapy with amphotericin B + flucytosine, and one example of a re-emergence of Cr.n. in the urogenital tract after a too short duration of treatment with amphotericin B + flucytosine are shown. For the exclusion of the survival of single CFUs of Cr.n. in the urogenital tract of males, quantities up to 1 l of urine for the combination of MFT and Staib agar are proposed. As a secondary observation, it was found that this diagnostic combination in addition to its primary purpose, can serve to detect the metabolic end products of the human body present in urine which may influence capsule formation of Cr.n. neoformans.
In a 33-year-old HIV-positive homosexual male suffering from unexplained headache, cryptococcosis was diagnosed in a progressive secondary stage. After treatment with the standard combination therapy of amphotericin B + flucytosine for 34 d, the patient was clinically symptom-free and discharged, upon his own request, from the hospital. He remained under ambulatory mycological control. After an interval of 65 d during which the urine had been free from Cryptococcus neoformans (Cr.n.), the fungus could not be isolated from urine but 3 X 10(5) CFUs/ml were found in the seminal fluid. Andrologically, teratospermia and hyposemia were present. There were no clinical signs in the genitourinary tract including the prostate. The significance of ecological niches for Cr.n. colonization of the genitourinary tract after antimycotic therapy is discussed. In such cases, in addition to cultural examination of urine for Cr.n. by the membrane filtration technique (MFT) and Staib agar, an additional cultural examination of seminal fluid is recommended. It is also proposed to pay more attention to Cr.n. in andrological examinations. Special regard should be given to a possible occurrence of Cr.n. in the seminal fluid of AIDS patients. In cytology of the seminal fluid, use of the Giemsa stain is unsuitable for the purpose of Cr.n. detection. For this reason, it should be supplemented by PAS staining.
It was demonstrated that the in vitro growth of a mucoid Escherichia coli strain from the urine of an AIDS patient could disturb the concurrent growth of Cryptococcus neoformans and the development of its brown colour effect (BCE) on Staib agar (syn. Guizotia abyssinica creatinine agar, bird seed agar, niger seed agar etc.) supplemented with penicillin + streptomycin. Owing to the supplementation with the triple antibiotic combination of penicillin + streptomycin + gentamicin and the resulting inhibition of E. coli growth, the formation of an intense BCE of the Cr. neoformans colonies after 3 d at 26 degrees C could be observed. On the same medium supplemented with this triple antibiotic combination 40 Cr. neoformans strains tested showed growth with an intense BCE after 3 d at 26 degrees C; but on Emmons' neutral Sabouraud's dextrose agar (NSDA) supplemented with the same triple antibiotic combination, inhibition of growth was found. For the examination of clinical specimens for Cr. neoformans contaminated with gram-negative rod-like bacteria, Staib agar supplemented with this triple antibiotic combination is proposed. Various antibiotic supplements to primary recovery media for fungi are discussed and ecological interrelations of bacteria and fungi are emphasized.
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Kidney failure and pneumonia by Aspergillus flavus and A. fumigatus were found in a 56-year-old woman who had received antibiotic and corticoid treatment to control high fever. Her bloody tracheal secretion was a suspension of granule-like spore-free colonies of both Aspergillus species. Hemorrhages in mucous membranes and skin suggested a hematogenous dissemination of the fungi. Aspergillus spores in the soil of ornamental plants were assumed to be responsible for the inhalatory infection. The kidney function normalized rapidly under treatment by amphotericin B plus flucytosine and hemodialysis performed eight times. After 29 days of antimycotic treatment (amphotericin B 463 mg, flucytosine 150 g), besides normalization of the kidney function, healing of the pneumonia and bleeding from skin and mucal membranes took place. One and a half years later kidney function and blood parameters were found to be normal. In cases of Aspergillus pneumonia and kidney failure, a combined treatment by hemodialysis and amphotericin B plus flucytosine is recommended. In addition, there is discussion of the general importance of uremia and its influence on the mycotic infection.
Cryptococcus neoformans was found as the causative agent of cryptococcosis in 7 (3.6%) out of 195 HIV-positive persons, most of them being hospitalized. These 7 persons included 6 homosexuals and 1 heroin addict. The examinations were performed at the Mycology Unit of the Robert Koch Institute in Berlin (West) between 1984 and 1986. The brown colour effect (BCE) of the C. neoformans colonies on Guizotia abyssinica creatinine agar (with 0.1% glucose) within 2-5 days at 26 degrees C facilitated the diagnosis of disseminated cryptococcosis. In all 7 cryptococcosis cases, the antigen of C. neoformans was detected in serum and CSF by the latex agglutination test. The initial titres ranged from 1:100,000 to 1:160 in the serum and from 1:1280 to 1:10 in the CSF. In comparison to the progressive stage of the infection with the involvement of the various organs and high antigen titres, the fungus may be detected at an early stage in the respiratory tract only where low antigen titres are observed. All the C. neoformans isolates were found to belong to the variety neoformans. Proposals for an effective control of cryptococcosis are made.
In two AIDS patients (homosexual men) microscopical demonstration of Cryptococcus neoformans in samples obtained by puncture of the liver (n = 1) and additionally of the spleen (n = 1) led to the diagnosis of systemic cryptococcosis. Using the India ink method capsulated Cryptococcus neoformans cells could also be detected in cerebrospinal fluid (CSF) and urine. Concomitant culture of the fungus from tracheal secretion, CSF, urine and faeces confirmed the diagnosis of a disseminated infection; the identification and germ count of C. neoformans was achieved by means of the differential-selective medium Guizotia-abyssinica-creatinine agar. The C. neoformans antigen titres in serum and CSF corresponded to the stage of the mycosis as detected by microscopy and culture. After a six-week course of treatment with amphotericin B and flucytosine (Ancotil), the fungus could no longer be isolated from the materials examined in one patient. Mycological monitoring aiming at the detection of C. neoformans in the tracheal secretions by means of the mentioned differential-selective medium is therefore recommended as a prophylactic measure in AIDS patients and persons at risk.
The risk for AIDS patients from Cryptococcus neoformans is outlined on the basis of a case report on a 28-year old male patient whose disease was complicated by cryptococcosis. Beside the description of the diagnosis of cryptococcosis (demonstration of the agent and its antigen), epidemiological associations (habitat of Cr. neoformans in fecal matter of birds) and the clinically, mostly not recognized, route of infection via the lungs is stressed. The effective therapy with the combination of amphotericin B and 5-flucytosine, which also in this case has been successful is described from the clinical and microbiological angles. Finally, proposals for the prevention of Cr. neoformans infections in AIDS patients and for a special mycological surveillance directed at this fungus are made.
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Cytostatic-treated persons and cases of severe hepatic failure under corticoid therapy are predisposed to disseminating Aspergillus infections. Constant exposure to Aspergillus spores may result in a fatal Aspergillus infection. The triad of hepatic failure, corticoid therapy and constant exposure to Aspergillus spores is described in a 70-year-old female patient. A painless icterus was clinically diagnosed as non-A non-B hepatitis, with a protracted cholestatic course. She had been treated with an oral corticoid preparation. After leaving the hospital at her own insistance when still in the icteric stage, severe pneumonia due to Aspergillus developed within 14 days; this was confirmed radiologically. The autopsy results showed unexpected infarction, similar to pneumonic foci, in all lobes and dissemination in the myocardium, stomach, kidneys and brain. The liver showed subacute dystrophy. Constant exposure to the conidia of A fumigatus came about as a result of the soil of potted ornamental plants in the patient's living room. The fungus could only be successfully cultured by putting infected tissue particles on Sabouraud dextrose agar; it was not possible by the common method of fractionated streaking.