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[Diagnosis of candida albicans mycosis in gynaecology by the germ tube test (author's transl)].

Experience with the germ tube test for the diagnosis of candida mycosis in gynaecology and dermatology is reported. Candida albicans forms germ tubes in human serum and other culture media at 37 degrees C within 2 to 3 hours. If the initial culture grows yeast, the germ tube test permits the exact identification of the yeast as candida albicans within three to five hours. The specificity of the test is approximately 99%. Of 3270 samples 934 grew yeast and 742 were candida albicans by the germ tube test. 6 samples with a negative germ tube test were identified as candida albicans by the Chlamydospore test. 125 samples showed a candida species, 45 samples showed a torulopsis species and 16 tests identified a trichosporon species. This test is valuable in the routine office practice of gynaecology because of its simplicity and speed.

Candidiasis, Cutaneous↗

A ten-year review of Candida sepsis and mortality in burn patients.

A retrospective analysis of Candida sepsis was carried out in 1722 burn patients admitted to this center from 1975 to 1984. Cultures were positive for Candida in 233 (13.5%) of these patients during their hospitalization. Candidemia was present in 70 (4.0%) of the 1722 patients. Of the 70 patients with candidemia, 38 (54%) died. However, only 11 patients (15.7%) died of Candida sepsis or mixed Candida and bacterial sepsis (less than 1% of the total patient population). The remaining 27 patients who had candidemia died of bacterial septicemia or organ system failure. The low incidence of Candida and the low incidence of mortality due to Candida was attributed to a comprehensive program of prevention, detection, and treatment. Early initiation of treatment with amphotericin B was an important aspect of the program.

Adult↗

How useful are symptoms in the diagnosis of Candida vaginitis?

Two hundred four female subjects symptomatic and asymptomatic for genitourinary disease were evaluated for Candida vaginitis. All were questioned regarding the presence or absence of a variety of symptoms related to the genitourinary system, including vaginal discharge, its color, if present, pruritus, dysuria, and the like. Additionally, all subjects were cultured for Candida by use of Sabouraud agar. Thirty-six subjects demonstrated cultures positive for Candida. Of 36 totally asymptomatic subjects, 4 had positive cultures, yielding a prevalence (18 percent) and asymptomatic positive rate (11 percent) consistent with those reported in the literature. All genitourinary symptoms individually and in combinations proved to be very poor predictors of the presence of Candida on culture. The study concludes that the diagnosis of Candida vaginitis cannot be made based on symptoms alone. A suggestion describing the office workup of Candida vaginitis is presented.

Adolescent↗

[Anatomo-clinical and therapeutic aspects of Candida albicans endophthalmitis].

Endophthalmia due to Candida has increased in incidence over the last few years, particularly in drug addicts. Two cases of severe bilateral Candida albicans endophthalmia are reported. Histological examination of the globes following treatment by amphotericin B i.v. and secondary vitrectomy demonstrated persistence of Candida, particularly in the preretinal membrane, in the first case. Treatment in the second case consisted of amphotericin B i.v. and 5 fluorocytosine, associated with vitrectomy and an intravitreal injection of 5 micrograms of amphotericin B. Fungal elements were absent on histological examination of the globes. Ocular Candida albicans lesions may be of exogenous origin but contamination arises more frequently from an endogenous source. The resulting endophthalmia leads to retinal nodules having a tendency to extend into the vitreous. Clinical features are fairly typical and diagnosis not a problem. General treatment consists of combined administration of amphotericin B and 5 fluorocytosine, but results are often disappointing as effective penetration of these compounds into the intraocular zone is not obtained. Early vitrectomy, when a vitreal lesion exists, offers many advantages such as the possibility to identify Candida in the removed aqueous humor. As histological examination shows persistence of Candida in the preretinal membrane, the vitrectomy should be combined with intravitreal injections of amphotericin B.

Aged↗

Measurement of Candida-specific blastogenesis: comparison of carboxyfluorescein succinimidyl ester labelling of T cells, thymidine incorporation, and CD69 expression.

Measurement of the T cell blastogenic response to Candida may be useful in the evaluation of patients with suspected immunodeficiency. The classic blastogenesis assay is based on uptake of [3H]thymidine by peripheral blood lymphocytes stimulated with Candida antigens for 5 days. An alternative approach involves staining peripheral blood lymphocytes with the intracellular fluorescent dye carboxyfluorescein succinimidyl ester (CFSE) and measuring mitotic activity by the successive twofold reductions in fluorescent intensity using flow cytometry (FCM). The two approaches were compared in 16 subjects who demonstrated various proliferative responses to Candida. FCM-derived indices all involved initial gating on CD3+ T cells and included 1) blastic transformation as measured by changes in light scatter, 2) cell division, measured by CFSE fluorescence, and 3) CD69 expression. A good correlation was found between [3H]thymidine uptake and CFSE-derived indices, irrespective of the analysis algorithm used to interpret CFSE division profiles. Furthermore, significant T cell proliferation occurred only in subjects who had had one or more symptomatic episodes of vaginal candidiasis whereas controls with no such history, and patients with chronic vaginal infection, showed minimal proliferation. The increase in proportion of CD69+ T cells in culture also correlated with the blastogenic response to Candida, but less well than mitotic indices. CFSE-derived indices of T cell blastogenesis to Candida are equivalent to [3H]thymidine-based assays and may allow useful laboratory distinction between subjects who have been exposed to and recovered from vaginal Candida infection, who have a strong proliferative response, from those with no exposure or chronic infection who demonstrate a poor response.

Antigens, CD↗

Reduced adherence of Candida to silane-treated silicone rubber.

Silicone rubber is widely used in the construction of medical devices that can provide an essential role in the treatment of human illness. However, subsequent microbial colonization of silicone rubber can result in clinical infection or device failure. The objective of this study was to determine the effectiveness of a novel silane-treated silicone rubber in inhibiting microbial adherence and material penetration. Test material was prepared by a combination of argon plasma discharge treatment and fluorinated silane coupling. Chemicophysical changes were then confirmed by X-ray photoelectron spectroscopy, contact-angle measurement, and atomic force microscopy. Two separate adherence assays and a material penetration assay assessed the performance of the new material against four strains of Candida species. Results showed a significant reduction (p < 0.01) of Candida albicans GDH 2346 adherence to silane-treated silicone compared with untreated controls. This reduction was still evident after the incorporation of saliva into the assay. Adherence inhibition also occurred with Candida tropicalis MMU and Candida krusei NCYC, although this was assay dependent. Reduced penetration of silane-treated silicone by Candida was evident when compared to untreated controls, plaster-processed silicone, and acrylic-processed silicone. To summarize, a novel silicone rubber material is described that inhibits both candidal adherence and material penetration. The clinical benefit and performance of this material remains to be determined.

Argon↗

Effects of granulocyte colony-stimulating factor and interferon-gamma on antifungal activity of human polymorphonuclear neutrophils against pseudohyphae of different medically important Candida species.

Polymorphonuclear neutrophils (PMNs) are the major host defense against pseudohyphae, the invasive form of Candida species. We studied the effects of granulocyte colony-stimulating factor (G-CSF) and interferon-gamma (IFN-gamma) on the PMN-induced damage of pseudo-hyphae of Candida albicans, Candida tropicalis, and Candida parapsilosis in vitro by using two antifungal assays: a modified limiting dilution assay and a colorimetric metabolic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. PMNs from healthy volunteers were incubated with either G-CSF (100-10,000 U/ml) or IFN-gamma (10-5000 U/ml) or buffer at 37 degrees C for 90 min and their capacity to damage nonopsonized pseudohyphae was then measured. C. tropicalis appeared to be the most susceptible species, whereas C. parapsilosis showed the highest rate of resistance to PMN damage. G-CSF (500-10,000 U/ml) and IFN-gamma (100-1000 U/ml) enhanced the antifungal activity of PMNs against C. albicans pseudo-hyphae (P < .01 and P < .05). Among the others, G-CSF enhanced PMN-induced damage of C. parapsilosis at concentrations 500-10,000 U/ml (P < .05), whereas it enhanced damage of C. tropicalis only at 10,000 U/ml (P < .01). IFN-gamma (100-1000 U/ml)-primed PMNs also caused augmented damage of C. parapsilosis (P < .05) but not of C. tropicalis at the same concentrations. Species-dependent differences exist in the responses of PMNs to Candida pseudohyphae and G-CSF as well as IFN-gamma are important immunomodulators of phagocytic host defenses against them.

Adult↗

Detection of Candida sp. mannan antigen by indirect ELISA-inhibition. In vitro crossed reactivity among mannans obtained from C. albicans A, C. albicans B, and C. tropicalis.

We have obtained mannans from four Candida species: C. albicans A, C. albicans B and C. tropicalis; anti-mannan sera against C. albicans A, C. albicans B and C. tropicalis were obtained by immunizing rabbits subcutaneously with the respective yeast extract. The efficacy of these sera in reacting with mannans obtained from three Candida sp. has been proven by indirect ELISA-inhibition. Any of three immune sera can be used to detect mannan antigen from the three Candida sp. tested. This confirms the existence of crossed reactivity and the possibility of detecting mannan antigen in serum from patients infected by different Candida sp., although we had only one immune serum and one Candida mannan.

Animals↗

Interactions of Candida albicans with bacteria and salivary molecules in oral biofilms.

The yeast Candida albicans coaggregates with a variety of streptococcal species, an interaction that may promote oral colonization by yeast cells. C. albicans and Candida tropicalis are the yeasts most frequently isolated from the human oral cavity and our data demonstrate that both these species bind to Streptococcus gordonii NCTC 7869 while two other Candida species (Candida krusei and Candida kefyr) do not. Adherence of C. albicans was greatest when the yeast had been grown at 30 degrees C to mid-exponential growth phase. For 21 strains of C. albicans there was a positive correlation between the ability to adhere to S. gordonii and adherence to experimental salivary pellicle. Whole saliva either stimulated or slightly inhibited adherence of C. albicans to S. gordonii depending on the streptococcal growth conditions. The results suggest that the major salivary adhesins and coaggregation adhesins of C. albicans are co-expressed.

Bacterial Adhesion↗

Comparison of three commercial media for direct identification and discrimination of Candida species in clinical specimens.

One hundred and ninety-two clinical specimens were used to compare the three chromogenic media Albicans ID, Candiselect, and CHROMagar Candida to a standard method using a Sabouraud-chloramphenicol agar medium and standard methods for identification of yeasts. The detection rates were 83.79, 83.24, 86.59 and 84.91% respectively. For the chromogenic media, the rates of direct identification (growth plus specific pigmentation) for Candida albicans were 56.50, 37.68 and 11.59% after 24 hours' incubation and 92.75, 91.30 and 88.57% after 72 hours' incubation respectively, with 100% specificity. Furthermore, CHROMagar Candida identified the five Candida tropicalis and the two Candida krusei strains detected after 48 hours' incubation.

Candida↗

Adherence in vitro of various Candida species to acrylic surfaces.

Adherence of microorganisms to the surfaces of the host is believed to be an initial and essential step in the production of infection. The objective of the present study was to compare the adherence in vitro of 17 isolates of various Candida species to acrylic surfaces. The results showed that all 17 Candida strains adhered to acryl. Generally, all Candida albicans strains were more adherent than the other strains of Candida species that were tested. Marked differences in the adherence ability of strains belonging to the same species were noted. Future studies to search for possible association between adherence of Candida strains to acryl and pathogenesis of denture stomatitis are planned.

Acrylic Resins↗

Palmitoyl carnitine, a lysophospholipase-transacylase inhibitor, prevents Candida adherence in vitro.

Candida adherence is poorly understood. The results of this study indicate that interactions of Candida with the lyso-forms of phospholipids may be one important attachment mechanism. C. tropicalis and C. albicans adhered to purified lysophospholipids immobilized on microtiter wells, as well as to a human laryngeal epidermoid carcinoma (HEp-2) cell line. Adherence to both lysophospholipids and HEp-2 cells was significantly reduced by palmitoyl carnitine, a lysophospholipase-transacylase inhibitor. Over time there was a positive correlation between Candida adherence and its transacylase activity. The data suggest that palmitoyl carnitine interferes with Candida adherence to lysophospholipids and the HEp-2 cell line by blocking the interaction between the Candida-associated transacylase enzyme receptor site and its lysophospholipid substrate ligand.

Acyltransferases↗

The isolation, identification and molecular analysis of Candida spp. isolated from the oral cavities of patients with diabetes mellitus.

Previous studies have shown a high incidence (77%) of isolation of Candida spp. from the oral cavities of patients with type 1 diabetes mellitus. The aim of the present study was to assess the prevalence of yeast in the oral cavities of patients suffering from type 1 and type 2 diabetes mellitus. The patients were classified according to the level of diabetic control (HbA1c), and further stratified on the presence or absence of dental prosthesis. Oral rinse samples were assessed for the growth of yeast and the degree of colonization. Oral isolates were defined to the species level by both phenotypic and novel molecular methods. The overall proportion (60%) of diabetic patients who had Candida spp. isolated from the oral cavity was similar to that previously reported. Local oral factors, such as the presence of dentures, seemed to have a greater influence than diabetic status on the amount and species of Candida isolated from the oral cavities of diabetic patients. Diabetic patients with dentures had more non-albicans Candida isolated from their mouths than dentate diabetic patients. Candida dubliniensis was isolated from diabetic patients and may have a predilection for dentate patients.

Adult↗

Alteration of humoral responses to Candida in HIV infection.

The objectives of this preliminary study were to determine the prevalence of oral candidal carriage and infection in a group of HIV-positive individuals and compare the humoral immune responses in serum and saliva in this group with a control group of HIV-negative subjects. Patients were examined clinically with particular reference to the presence of candidal lesions and oral swabs taken to identify carriers. Venous blood and whole saliva were obtained for estimation of total and anti-Candida antibody levels. Pseudomembranous candidiasis was the commonest clinical variant in HIV-positive individuals. Candida albicans was the commonest species isolated in both groups. Increased levels of anti-Candida IgG were found in both serum and saliva of HIV-negative individuals who were either carriers of Candida species or had clinical candidiasis. This was associated with a reciprocal fall in anti-Candida IgA. Similar trends were seen in HIV-positive individuals in association with candidal carriage and infection, although the changes were more marked.

Antibodies, Fungal↗

Vertebral osteomyelitis due to Candida species: case report and literature review.

Candida species uncommonly cause vertebral osteomyelitis. We present a case of lumbar vertebral osteomyelitis caused by Candida albicans and review 59 cases of candidal vertebral osteomyelitis reported in the literature. The mean age was 50 years, and the lower thoracic or lumbar spine was involved in 95% of patients. Eighty-three percent of patients had back pain for >1 month, 32% presented with fever, and 19% had neurological deficits. The erythrocyte sedimentation rate was elevated in 87% of patients, and blood culture yielded Candida species for 51%. C. albicans was responsible for 62% of cases, Candida tropicalis for 19%, and Candida glabrata for 14%. Risk factors for candidal vertebral osteomyelitis were the presence of a central venous catheter, antibiotic use, immunosuppression, and injection drug use. Medical and surgical therapies were both used, and amphotericin B was the primary antifungal agent. Prognosis was good, with an overall clinical cure rate of 85%.

Adult↗

Differential inhibitory effects of protoberberines on sterol and chitin biosyntheses in Candida albicans.

The anti-Candida potentials of 12 Korean medicinal plants were explored: methanol extracts from Coptis rhizoma and Phellodendron amurense caused significant inhibition of growth of Candida albicans, Candida glabrata, Candida krusei and Candida parapsilosis. The predominant active components of the extracts were the protoberberines berberine and palmatine; the most potent inhibition of growth was exhibited by berberine on C. krusei (MIC <4 mg/L) and palmatine on C. parapsilosis (MIC 16 mg/L). Both berberine and palmatine inhibited the in-vivo rate of incorporation of L-[methyl-14C]methionine into C-24 of ergosterol in C. albicans (50% inhibition concentration (IC50 values), 25 microM and 300 microM, respectively); this result suggests that sterol 24-methyl transferase (24-SMT) is one of the cellular targets for the antifungal activity of the protoberberines. In-vitro 24-SMT activity in microsomes from the yeast growth form of C. albicans was inhibited by both berberine (inhibition constant (Ki) 232 microM) and palmatine (Ki 257 microM) in a non-competitive manner; inhibition of 24-SMT was more marked for the mycelial form than for the yeast growth form of this organism. Palmatine inhibited chitin synthase from both the yeast and mycelial growth phases of C. albicans in a non-competitive manner (Ki 780 microM). The effects of protoberberines, extracted from established medicinal plants, on both sterol and cell wall biosyntheses in pathogenic fungi indicate that the potential of these compounds, or their semi-synthetic derivatives, as a novel class of antifungal agents should be investigated more fully.

Amphotericin B↗

Distribution of a corticosteroid-binding protein in Candida and other fungal genera.

Using [3H]corticosterone as a probe, corticosteroid-binding protein (CBP) was detected in eight out of eight isolates of Candida albicans, of both A and B serotypes. The apparent dissociation constant (Kd) in the various isolates ranged between 8 and 19 nM; the binding capacity varied from 122 to over 2400 fmol (mg cytosol protein)-1. There was no correlation between the amount or affinity of CBP and isolate virulence for murine hosts. Further analysis revealed demonstrable CBP in six out of six Candida species other than C. albicans. One isolate of C. tropicalis has been identified which fails to bind [3H]corticosterone. Saccharomyces cerevisiae, Neurospora crassa and Paracoccidioides brasiliensis also failed to bind [3H]corticosterone. Preliminary attempts were made to determine functions mediated by CBP in Candida, but in vitro growth, phase conversion and glucose oxidation by Candida were unaffected by the addition of a variety of steroid hormones. These data indicate that the presence of CBP in Candida does not correlate with either virulence or serotype. The physiological significance of CBP remains to be determined.

Candida↗

Activation of the alternative (properdin) pathway of complement by Candida albicans and related species.

Accumulations of neutrophilic granulocytes within the epidermis and beneath the stratum corneum of the skin are a prominent histologic feature of experimental and clinical candidiasis. The mechanism of cell accumulation was studied by standard chemotactic methods. Suspensions of viable or heat-killed Candida sp caused marked chemotaxis of human neutrophils in fresh serum. Culture supernatants of Candida sp were not chemotactic. Chemotaxis was dependent upon fresh serum, and could be abolished by heating the serum to 56 degress C for 30 min, suggesting that interaction of these organisms with a heat-labile serum factor generated a chemoattractant. Incubation of Candida sp with fresh human serum resulted in the conversion of the third component of complement and properdin factor B, as measured by immunoelectrophoresis. Conversion did not occur in serum chelated with EDTA, or heated to 50 degress C for 30 min (to destroy factor B). Conversion was present in serum chelated with EGTA (to deplete calcium), or genetically deficient in the fourth component of complement. By contrast, the three components of the kinin-forming system (Hageman factor, prekallikrein, high-molecular-weight kininogen) were not activated by Candida sp. We suggest that Candida sp do not release a chemotactic substance but, in the presence of serum, activate the alternative pathway of complement, generating chemotactic factors.

Candida↗