Deep-seated Candidiasis Research Articles

Vital staining of fungal elements in deep-seated mycotic lesions during experimental murine mycoses using the parenterally applied optical brightener Blankophor.

Optical brighteners of the diaminostilbene type are fluorescent dyes which are popular diagnostic tools in the mycology laboratory. While these dyes are conventionally used for the in vitro diagnosis of mycoses, their low toxicity and chemical reactivity have led us to investigate their potential use for in vivo staining of fungal elements in mycotic tissue. In mice we have established deep-seated candidiasis, cryptococcosis, aspergillosis and zygomycosis, as well as coccidioidomycosis, histoplasmosis and blastomycosis. After establishment of infection, which mostly required immunosuppression, a single dose of 100 microl of an aqueous solution (2.2 x 10(-4) M) of the optical brightener Blankophor P fluessig (4,4'-Bis [(4-anilino-6-substituted-1,3,5-triazine-2-yl) amino] stilbene-2,2'-disulfonic acid) was injected by the tail vein and the animals were sacrificed 1 h later. Sections of freshly prepared target organs were directly subjected to epifluorescence microscopy using an appropriate filter kit. In most cases, fluorescent fungal elements could be detected in the murine tissue. There was little evidence for uptake of the dye by non-infected tissues. It is suggested that radioactive labeling may render parenteral Blankophor suitable for radiographic localization of deep-seated mycotic foci in the host.

Rapid detection of Candida albicans in clinical samples by DNA amplification of common regions from C. albicans-secreted aspartic proteinase genes.

Laboratory diagnosis based on genomic amplification methods such as PCR may provide an alternative and more sensitive method than conventional culture for the early detection of deep-seated candidiasis, an increasing cause of morbidity and mortality among immunocompromised patients. A novel method of DNA extraction from clinical samples based on treatment with proteinase K and isolation of DNA on a silica membrane was developed. The targets used for DNA amplification were the Candida albicans-secreted aspartic proteinase (SAP) genes, a multiple-gene family of at least seven members in C. albicans. A single pair of primers was designed in order to detect six of these SAP genes and, subsequently, to increase the sensitivity of the test. Detection of the PCR product by enzyme-linked immunosorbent assay was found to be as sensitive as Southern blotting with an SAP-labeled probe. The sensitivity of the assay was 1 cell/ml from serially diluted Candida cultures and 1 to 4 cells/ml from seeded blood specimens. The sensitivity and specificity of the present assay were tested in a retrospective study performed blindly with 156 clinical samples and were 100 and 98%, respectively, compared with the results of culture. For the subset of blood culture samples (n = 124), the sensitivity and the specificity were 100%. The two false-positive PCR samples came from patients treated with azole antifungal agents, indicating that PCR was probably able to detect damaged organisms that could not be recovered by culture.

The studies on tuberculosis present research ideas for medicine

I started my medical career as medical mycologist following the first case of deep-seated candidiasis in Japan. On the other hand I have been seeing many patients with tuberculosis as well as respiratory diseases. In my sense the seeing of mycobacterial infections may still remain lot of clinical, biological or immunological research ideas in future. I did the studies on muramyl dipeptide (MDP) which derived from cell wall substance of mycobacteria. MDP seemed to be one of the enhancing immunomodulator for host defense mechanism in particular in the immunocompromised host.

Decrease in Candidemia Rates Associated with Empiric Use of Amphotericin B and Fluconazole for Nosocomial Fever of Unknown Origin

Since 1986, patients with nosocomial fever of unknown origin (NFUO) at Methodist Central Hospital (Memphis) have been initially treated with antibacterial antibiotics if there was a likelihood of infection. If there was no response to therapy and noninfectious causes for the NFUO were eliminated, treatment was then initiated with antifungal agents, notably amphotericin B and fluconazole. Most patients so treated were nonneutropenic and in the intensive care unit (ICU). A retrospective analysis using hospital data collected between 1987 and 1994 suggested that hospitalized patients had an increasing severity of illness during the study period yet a decreasing incidence of candidemia, particularly after 1990 when fluconazole was approved and used frequently for NFUO. From 1988 to 1990, the rate of candidemia per 1000 ICU admissions was 5.56; while from 1991 to 1994, the rate was 2.64 per 1000 ICU admissions (P <.0001). Since the data suggest that early empiric therapy of NFUO with antifungal agents is useful when a patient is at high risk for deepseated candidiasis, a prospective study to evaluate this method of clinical management is indicated.

Retrospective analysis of deep-seated candidiasis among cases autopsied between 1982 to 1991

Candida is present in the flora of the oral cavity, skin, intestinal tract and vagina, and is also known to be an opportunistic pathogen. Infection with this fungus has been increasing annually along with wide spread use of broad-spectrum antimicrobial agents. The subjects included 95 patients (48 males and 47 females) who had been diagnosed as having had deep-seated candidiasis, among patients autopsied between 1982 to 1991. In regard to annual changes in deep-seated candidiasis, the incidence reached a peak in the 1985 to 1988 period, and thereafter decreased. The number of cases with leukemia as the underlying disease was the largest, 36 (37.9%), followed by malignant lymphoma in 10, and aplastic anemia 5. The number of cases with infection of the stomach was largest, 42 (44.2%), followed by the esophagus in 33 (34.7%), the lung and kidney. The cases with deep-seated candidiasis showed low values of or level of lymphocyte, hemoglobin, CRP, total protein and cholesterol and high values or levels of LDH, urea N, creatinine and total bilirubin. Cases with marked decrease in neutrophils showed no regional infiltration of inflammatory cells in any of the organs infected with Candida. Cases with disseminated candidiasis showed vascular invasion by Candida. The laboratory findings also showed that most of the cases had been undernourished and had high values of CRP which supports the presence of inflammation. Common sites of infection are the esophagus, stomach, and intestinal tract. In the presence of granulocytopenia and immunodeficiency, tissue invasion become severe and associated with vascular invasion.

Surveillance and treatment of liver transplant recipients for candidiasis and aspergillosis.

Between June 1988 and May 1991 88 orthotopic liver transplants and 1 liver and pancreas transplant were performed at the Liver Transplantation Department of the Ospedale Maggiore of Milan. All the patients underwent mycological surveillance and received antifungal prophylaxis with oral amphotericin B (6000 mg/day) or oral or intravenous fluconazole (200 mg/day) from the time of their transplant. The incidence of Candida colonization was 67%. Fluconazole was superior to oral amphotericin B in the treatment of C. albicans colonization (9/9 vs 6/15), but less effective in the treatment of colonization by other Candida spp. (0/3 vs 3/3). Deep-seated candidiasis developed in 5 patients, caused by C. albicans in 4 cases and C. krusei in 1. C. albicans infection resolved rapidly with fluconazole in 2 subjects, with intravenous amphotericin B alone in 1, and with amphotericin B plus flucytosine in the other. On the contrary, C. krusei infection did not respond to treatment with amphotericin B combined with flucytosine. Aspergillosis was diagnosed in 11 patients, of whom 4 died from invasive aspergillosis, despite 15 and 26 days of amphotericin B treatment in 2. In another patient invasive aspergillosis, diagnosed a few hours before retransplantation, improved with liposomal amphotericin B, but this man died from cytomegalovirus infection one month later. Aspergillosis was eradicated by itraconazole in 4 other patients and by topical amphotericin B in 2 whose infection was localized to surgical wound.

Candidiasis visualised by proteinase-directed immunofluorescence

The secretory aspartic proteinases of Candida albicans and C. tropicalis are potential factors for virulence produced during infection. By indirect immunofluorescence, we have demonstrated proteinase antigen on elements of both species in deparaffinized tissue sections derived from clinical cases of mucosal and deep-seated candidiasis. Occasionally, we observed a halo of fluorescence in the close vicinity of candidal cells, which may reflect secretion of the enzyme. In kidneys, a ring of amorphous fluorescent material surrounding candidal colonies may illustrate alkaline denaturation of secreted enzyme within a pH gradient, which is generated by the fungus. Our findings support the view that candidal proteinase may be a diagnostically relevant antigen.

Fluconazole

Fluconazole is a bis-triazole antifungal drug with novel pharmacokinetic properties (metabolic stability, relatively high water solubility) which contribute to its therapeutic activity. Clinical experience is limited to a relatively small number of mycoses and, as might be expected at this early stage of development, optimal dosage and duration of treatment for some serious mycoses is not yet established. Further study to evaluate higher dosages and to establish the efficacy of fluconazole relative to more established antifungal agents is required. In patients with oropharyngeal or oesophageal candidiasis, fluconazole produces rapid relief and eradicates the yeast in 50 to 90% of patients. Relapse of oral infection is common in chronically immunocompromised patients regardless of the antifungal used, and adequate primary therapy plus long term prophylaxis appears necessary in patients with AIDS. A single oral dose of fluconazole was comparable to standard topical azole therapy in women with acute vaginal candidiasis. Preliminary reports of success against deep-seated candidiasis are encouraging; moreover, experience in noncomparative clinical trials suggests that fluconazole 200 to 400mg once daily resolves infection in the majority of seriously ill patients. Clinical improvement has been reported in a few cases of pulmonary Aspergillus infection but the overall efficacy of conventional dosages of fluconazole in this mycosis has not been as impressive. Early experience in coccidioidosis, predominantly meningitis, suggests a beneficial clinical effect with oral fluconazole in this difficult to treat mycosis but relapse remains a problem. Fluconazole is a promising treatment of cryptococcal meningitis. The rate of clinical resolution and eradication of Cryptococcus neoformans from cerebrospinal fluid has been similar between fluconazole and amphotericin B treatment groups in comparative trials. Comparative trials of maintenance therapy indicate a similar low rate of relapse among patients given oral fluconazole once daily and intravenous amphotericin B once weekly. However, these results are preliminary and further study is required. Fluconazole has been well tolerated to date but wider clinical experience is needed, especially with regard to the rate occurrence of hepatotoxicity and exfoliative skin reactions. The promising clinical response of patients with various forms of candidiasis or cryptococcosis--together with convenient administration regimens--recommends fluconazole as a useful addition to currently available systemic antifungal therapies, in particular for the treatment of mycoses in patients with AIDS.

Detection of an antibody response in immunocompetent patients with systemic candidiasis orCandida albicans colonisation

Tests to detect circulating antibodies to Candida albicans antigens were performed in sera from 27 immunocompetent patients, 15 of whom had deep-seated candidiasis and 12 of whom were colonised by Candida albicans. For the diagnosis of deep-seated candidiasis in patients with either deep-seated candidiasis or Candida albicans colonisation, counterimmunoelectrophoresis had a sensitivity of 87% and a specificity of 75%. Using immunoblotting it could be shown that antibodies to 35K, 47K, 68K and 88K antigens of Candida albicans occurred more frequently in patients with deep-seated candidiasis than in colonised patients. The presence of dense bands in immunoblots representing antibodies against the 47K and/or 68K antigen served to discriminate significantly between deep-seated and superficial candidiasis (p less than 0.05).

血液疾患に合併した真菌症とくに深在性カンジダ症の検討

Deep-seated mycosis is prominently increasing as a terminal infection in compromised hosts with malignant blood disorders or malignant tumors. Moreover, localized candidal abscess of visual organs has recently been reported in several laboratories. We investigated the occurrence of deep-seated mycosis in 105 autopsied cases with blood disorders in our clinic from 1980 to 1987. Forty-four of those cases had died of various infections, and 80% of them were fungal infections. More than half of the fungal infections were aspergillosis. Deep-seated candidiasis was recognized in 10 cases, 6 of which were systemic candidiasis, the average duration of neutropenia below 500/mm3 was 19.7 days that of lymphopenia was 36.5 days. Two cases were complicated with GI-tract ulcer, and involved with hepatic candidiasis. On the other hand, in the 4 cases of localized candidial abscess, the duration of neutropenia was 58.5 days and that of lymphopenia was 28.8 days. These four cases were complicated with GI-tract ulcer. Histologically, Candida spp. were recognized at the bottom of the ulcer and invasion by inflammatory cells or tumor cells was found in the portal vein. We surmised that GI-tract ulceration is a very important complication of hepatic candidiasis or liver abscess, and the occurrence of localized candidiasis seems to depend on the duration and severity of neutropenia.

A serum mannan-binding protein and candidiasis.

A mannan-binding protein purified from rabbit serum agglutinated Saccharomyces cerevisiae in a Ca2+-dependent manner. Its binding to related carbohydrate ligands is reduced either by yeast mannan at 10 ng ml-1 or serum from patients with suspected deep-seated candidiasis. A role for mannan-binding protein in diagnosis and pathogenesis is considered.

Diagnostic Value of Monitoring Kinetics of Antibody Responses in Candidiasis by a Solid-Phase Radioimmunoassay

A solid-phase radioimmunoassay was used to monitor antibody binding to cytoplasmic antigens of Candida albicans in sera from patients with superficial or deep-seated candidiasis over extended observation periods. Healthy persons showed a constant level of Candida antibodies. Patients with superficial candidiasis or colonization generally had a slow, but significant rise; patients with presumptive and proven systemic candidiasis, however, developed rapid rises (up to forty-fold) of antibody levels within a few days. This method permitted the kinetics of antibody response to be followed and, thus, appeared to aid in the differentiation among the various forms of candidiasis.

Diagnostic value of monitoring kinetics of antibody responses in candidiasis by a solid-phase radioimmunoassay.

A solid-phase radioimmunoassay was used to monitor antibody binding to cytoplasmic antigens of Candida albicans in sera from patients with superficial or deep-seated candidiasis over extended observation periods. Healthy persons showed a constant level of Candida antibodies. Patients with superficial candidiasis or colonization generally had a slow, but significant rise; patients with presumptive and proven systemic candidiasis, however, developed rapid rises (up to forty-fold) of antibody levels within a few days. This method permitted the kinetics of antibody response to be followed and, thus, appeared to aid in the differentiation among the various forms of candidiasis.

Increasing the predictive value positive of the precipitin test for the diagnosis of deep-seated candidiasis.

Three hundred fifty human sera were tested by double immunodiffusion, crossed-line electrophoresis, and crossed immuno-affinoelectrophoresis with a concanavalin A intermediate gel for precipitating antibodies to antigens present in cytoplasmic extracts of Candida albicans. Sera from 48 of 287 hospitalized patients at risk of invasive candidiasis contained precipitating antibodies to Candida antigens. Of these 48 sera, 27 had precipitating antibodies only to cell-wall antigens present in the cytoplasmic extract, and 21 sera had precipitating antibodies to both cytoplasmic and cell-wall antigens. The latter sera came from patients who were 2.5 times as likely to have deep-seated candidiasis as those patients with precipitins exclusively to cell-wall antigens. Sera from seven of 22 patients with vaginal candidiasis and 10 of 41 patients with other fungal infections had precipitating antibodies to C. albicans cell-wall antigens; only two of these sera also contained precipitating antibodies to the cytoplasmic antigens. Crossed immunoaffinoelectrophoresis with concanavalin A reduced the number of false-positive results and increased the predictive value positive of the precipitin test for deep-seated candidiasis from 31% to 71%.

Increasing the Predictive Value Positive of the Precipitin Test for the Diagnosis of Deep-seated Candidiasis

Increasing the Predictive Value Positive of the Precipitin Test for the Diagnosis of Deep-seated Candidiasis

Immunologic Responses to Candida Albicans

Abstract Passive transfer of immune serum gave a significant degree of protection against deep seated candidiasis in mice. Repeated attempts to transfer resistance by the transfer of sensitized lymphoid cells gave negative results, even though cutaneous delayed hypersensitivity was transferred by the cells. The results suggest that cell-mediated immunity is not of primary importance in this model of murine candidiasis, and that humoral immunity contributes to protection.

Pneumocystis and fungal infection in patients with malignancies

The alarming increase in infections resulting from fungi and Pneumocystis among patients with malignancies has prompted the organization of a number of workshops to explore the magnitude of the problem and to elicit suggestions for approaching diagnosis, treatment and prevention of infection with these opportunistic pathogens. Our knowledge of the source and means of spread of organisms like Aspergillus and Pneumocystis, and the methods of preventing infection by them have advanced only little in spite of the acknowledged high incidence of these infections in major medical centers in the United ‘States during the last decade. Premortem diagnosis of infection with Pneumocystis and those fungi causing the greatest incidence of morbidity and mortality in these patients rarely is accomplished without invasive procedures; frequently, even when the diagnosis is suspected premortem it may be established solely at autopsy. We shall review briefly our own experience with deep seated (most often disseminated) candidiasis and Pneumocystis infection in tumor bearing patients. For a review of the literature, the reader is referred to other references. (1.6.7.9.11.12.14-16) Aspergillus has been a

Assessment of germ tube dispersion activity of serum from experimental candidiasis: a new procedure for serodiagnosis.

Germ tubes of Candida albicans and C. stellatoidea clump in normal serum but disperse in serum from animals infected with either species of Candida. A new procedure for the assessment of grades of germ tube dispersion activity of serum is presented; this procedure is to count the number of freely dispersed germ tubes in test serum into which a definite number of yeast-type C. albicans has been inoculated. The relationship between the serum activity and macroscopic lesions caused by candidal infection is observed, indicating the possibility of applying the phenomenon to the serodiagnosis of deep-seated candidiasis. The specificity and sensitivity of the test are also examined.

Four Cases of Deep-Seated Candidiasis of Pharynx and Larynx

Four Cases of Deep-Seated Candidiasis of Pharynx and Larynx

Deep-seated Candidiasis

Deep-seated Candidiasis