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PubMed1994

Transfer RNA profiling: a new method for the identification of pathogenic Candida species.

A new molecular taxonomic method applicable to the identification of medically important Candida species and other yeast species has been developed. It is based on the electrophoretic pattern of total tRNA samples (a 'tRNA profile') isolated from Candida species and generated using high-resolution semi-denaturing urea-polyacrylamide gel electrophoresis and methylene blue staining. Species-specific tRNA profiles for the species C. albicans, C. tropicalis, C. parapsilosis, C. guilliermondii, C. glabrata and Pichia guilliermondii were obtained. Detailed studies with the major human pathogen of the Candida genus, C. albicans, demonstrated that the tRNA profile for a given species was both reproducible and strain-independent; seven different C. albicans strains generated identical tRNA profiles. Minor strain-specific heterogeneities in the tRNA profiles of C. guilliermondii and C. parapsilosis were detected, but in neither case did they significantly alter the species-specific diagnostic tRNA profile. The potential of this method in clarifying taxonomic anomalies was demonstrated by the finding that Type I and Type II strains of C. stellatoidea generate very different tRNA profiles, with that of a Type II strain being identical to the C. albicans tRNA profile. This method offers a number of advantages over current electrophoretic karyotype methods for species identification, both within the Candida genus and with yeast species in general.

Candida↗
PubMed1994

Heterogeneous secretory response of phagocytes from different anatomical districts to the dimorphic fungus Candida albicans.

In the present study, we have examined the ability of phagocytes from different anatomical districts to discriminate between the two morphogenetic forms of Candida albicans. We have demonstrated that resident peritoneal macrophages (RP-M phi) and thioglycollate-elicited peritoneal macrophages (TP-M phi) were able to distinguish between the hyphal (H-Candida) and the yeast (Y-Candida) form of the fungus, since TNF production was observed only upon exposure of RP-M phi and TP-M phi to H-Candida. In contrast, splenic macrophages (S-M phi), bone marrow-derived macrophages (BM-M phi) and polymorphonuclear neutrophils (PMN) did not discriminate between the two forms because S-M phi and PMN produced TNF regardless of the morphogenetic status of the fungus, while BM-M phi did not. Under the same experimental conditions, we failed to observe IL-1 production from any of the phagocytic cell populations examined, with the exception of PMN. This implies that the interaction between phagocytes and C. albicans triggers differential secretory responses depending upon the morphogenetic status of the fungus and the anatomical localization of the immune cells.

Animals↗
PubMed1994

Glucocorticoid-induced alterations of monocyte defense mechanisms against Candida albicans.

The mechanisms of glucocorticoid-induced immunosuppression of human monocytes for the defense of Candida albicans were examined in this in vitro study. Dexamethasone at pharmacological concentrations (10(-7)-10(-5) M) dose-dependently attenuated growth inhibition of C. albicans by resident monocytes. In interferon-gamma (IFN-gamma)-primed monocytes, fungal growth inhibition was maximal and not altered by dexamethasone. Similarly, phagocytosis and killing of Candida by monocytes were not affected by steroids. To study the underlying mechanisms, we found that Candida-induced synthesis of tumor necrosis factor-alpha (TNF-alpha) by monocytes was completely abrogated by dexamethasone. Substitution of TNF-alpha to dexamethasone-treated monocytes fully reversed the alterations of growth inhibition of C. albicans induced by steroids. These findings suggest that glucocorticoids affect the growth control of Candida by monocytes indirectly via suppression of the formation of TNF-alpha, which is required as an autocrine cofactor for full monocyte activation.

Candida albicans↗
PubMed1994

Different events involved in the induction of macrophage tumor necrosis factor by Candida albicans and lipopolysaccharide.

Using an in vitro experimental model, we have recently demonstrated that Candida albicans in its hyphal form (H-Candida), similarly to lipopolysaccharide (LPS), enhances tumor necrosis factor (TNF) secretory response in the cloned macrophage (M phi) population ANA-1. Here we show that H-Candida and LPS each differ in their requirements for intact protein kinase functions, susceptibility to 0.4-microns micropore-size membranes, and sensitivity to polymyxin B. These results, together with the synergistic effect occurring between H-Candida and LPS in inducing TNF response, indicate the existence of different receptor(s) and/or signal-transduction pathway(s) through which the two stimuli act.

Animals↗
PubMed1996

Synergy of human neutrophils with fluconazole in killing Candida species.

The killing of Candida species by human neutrophils in a long-term 24-h assay and possible synergy with fluconazole (FCZ) for killing was investigated. The test medium (TM) consisted of RPMI-1640, penicillin and streptomycin (P/S), and 10% fresh autologous serum. TM alone was highly fungistatic for Candida species compared to TM without serum. When neutrophils were cocultured in TM with Candida species for 24 h the inoculum colony-forming units (CFU) were always significantly reduced (killing) by 58 to 99%. FCZ was tested over a range of 1-500 micrograms/ml, and though almost always fungistatic itself, it synergized with neutrophils for significantly increased killing of C. albicans (isolate Sh27) (P < 0.01) and C. albicans (isolate 94-20) (P < 0.05). Killing of non-albicans Candida species was so efficient in the absence of FCZ that demonstration of synergy with FCZ was difficult.

Antifungal Agents↗
PubMed1986

In vitro modification of Candida albicans invasiveness.

Candida albicans produces germ-tubes (GT) when it is incubated in animal or human serum. This dimorphism is responsible for its invasive ability. The purpose of the present paper is (1) to evaluate the ability of rat peritoneal macrophages to inhibit GT production of ingested Candida albicans, obtained from immunized rats and then activated in vitro with Candida-induced lymphokines; (2) to determinate any possible alteration of phagocytic and candidacidal activities. The phagocytes were obtained from rats immunized with viable C. albicans. Some of them were exposed to Candida-induced lymphokines in order to activate the macrophages in vitro. The monolayers of activated, immune and normal macrophages were infected with a C. albicans suspension during 4 hr. Activated macrophages presented not only the highest phagocytic and candidacidal activities but a noticeable inhibition of GT formation and incremented candidacidal activity.

Animals↗
PubMed1989

Ethanol inhibition of Saccharomyces and Candida enzymes.

Ethanol inhibition of several hydrolases (sucrase, maltase, trehalase, melezitase and cellobiase) has been measured in both highly ethanol-tolerant Saccharomyces strains (R) and in Candida strains less tolerant to ethanol (S). Cells were either grown in the presence of ethanol and the activities of the enzymes measured without preincubation in this alcohol ("in situ" inhibition assay), or the culture was grown in the absence of ethanol and the activities of the enzymes were determined after preincubation and in the presence of this compound ("in vitro" inhibition assay). Ethanol inhibition (Ki values) of sucrase, maltase, trehalase, and melezitase was quite different for these different enzymes in the same strain (R or S), but similar for the same enzyme in different strains (R and S). The Ki values for cellobiase, which is absent from the R strain, were higher when induced than at the basal level and higher in in vitro assays than in in situ assays. This suggests that the inhibition observed in situ is mainly the result of an inhibition of other proteins related to cellobiase (i.e., those involved in its synthesis) but not a direct inactivation of the enzyme by ethanol. Accordingly, when hybrids between Saccharomyces (R) and Candida (S) strains were constructed by protoplast fusion, and cellobiase was measured in the parental Candida strain and some of the hybrids, there was an increase in the Ki values in the in situ assays from 2.25% ethanol in Candida to 5.5% in some of the hybrids.

Candida↗
PubMed1992

Isolation of auxotrophic mutants of the methylotrophic yeast Candida boidinii and determination of its ploidy.

Auxotrophic mutations in the methylotrophic yeast strain Candida boidinii 11Bh were induced by different mutagens and their combinations (nitrosoguanidine, UV light, HNO2+UV). Majority of the mutants obtained carried defects in histidine, arginine, proline and/or adenine biosynthetic pathways. His- mutants were distributed into four complementation groups using the protoplasts fusion technique. Ploidy determination of Candida boidinii 11Bh was performed by measuring its DNA content and by following its survival after chemical mutagens treatment. The DNA content of this strain was found to be similar to that of a Saccharomyces cerevisiae diploid strain. Also the kinetics of survival of Candida boidinii cells indicate that Candida boidinii 11Bh is a diploid.

Candida↗
PubMed1996

Role of Candida in indirect pathogenesis of antibiotic associated diarrhoea in infants.

One hundred and thirty seven isolates of Candida species were isolated from antibiotic associated diarrhoea cases and were examined to study the role of Candida in the pathogenesis of diarrhoea in infants. The quantitative estimation of yeast population by simple gram stain smear revealed more than 70% of the cases had 3+ score. The isolates further screened for detection of beta-lactamases. Among the isolated Candida sp, beta-lactamases was secreted by C. albicans, C. tropicalis, C. krusei and C. parapsilosis. Further, 46% of the Candida isolates were found to be produced 741-1110 mU/ml of beta-lactamases, suggesting that these enzyme would inactivate penicillin group of drugs and cause failure in the therapy directed against other diarrhoegenic bacteria.

Anti-Bacterial Agents↗
PubMed1994

Significance of modes of adherence in esophageal Candida albicans.

Although esophageal candidiasis is the most common form of Candida infection in the gastro-intestinal tract, little attention has been directed toward determining the mechanism of its infection. We have already clarified the existence of four modes of adherence of Candida albicans to the esophagus; attachment, subepithelial cell insertion, cavitation, and invasion. This study was undertaken to clarify the significance of each of these modes. Scanning electron microscopic observations were made of esophageal specimens from 8-week-old rabbits infected with Candida albicans IFO 1060. In this study, attachment and subepithelial cell insertion were found to be the most frequent modes of adherence. Cavitation occurred following subepithelial cell insertion, while invasion occurred following attachment and subepithelial cell insertion. These results suggest that attachment and subepithelial cell insertion play the most important role in the initial stage of adherence. The ratios of these modes for living yeast cells were similar to those for dead yeast cells and beads. This suggests that Candida albicans can gain a foothold on the esophageal epithelium solely by physical contact, after which colonization occurs.

Animals↗
PubMedDate not supplied

Correlation between antifungal susceptibility testing of Candida isolates from patients with HIV infection and clinical results after treatment with fluconazole.

In an open-label controlled study 23 HIV-infected patients (CDC IV A-E) with documented oropharyngeal candidosis were treated with 100 mg fluconazole orally over 5 days (53 episodes; 1-6 treatments/patient). Efficacy data were compared with a control group of 21 patients who received treatment for 10-21 days with 100 mg fluconazole for candidosis. Candida isolates were repeatedly recovered from patients before and after treatment with fluconazole and antifungal susceptibility testing (microbroth-dilution) was done. Inoculum size, medium pH, incubation time and temperature were standardized. Up to 85% of patients responded to therapy clinically and mycologically. Candida albicans was the most important yeast (86%) isolated from cultures of oral washings. In 90% of C. albicans isolates MIC to fluconazole were low (< or = 1.56 mg/l). Primary resistance to fluconazole was not seen, but secondary resistance occurred in two cases clinically and in vitro (MIC > or = 25 mg/l). Short treatment for 5 days was as successful as for 10 to 21 days without leading to significantly more recurrences of oral candidosis in these patients. Selection of Candida spp. other than C. albicans (e.g. Candida krusei, Torulopsis glabrata) under repeated fluconazole treatment occurred rarely. One patient developed clinical signs of chronic recurrent candidiasis, where only C. krusei could be cultured repeatedly.

AIDS-Related Opportunistic Infections↗
PubMedDate not supplied

Fluconazole in Candida albicans sepsis during pregnancy: case report and review of the literature.

Candida sepsis during pregnancy is a rare but life-threatening complication of infection with Candida albicans. In contrast to the situation with other antimicrobial agents, there exists only limited experience with systemic antifungal therapy during pregnancy. A recent report focuses on amphotericin B treatment in systemic fungal infection during pregnancy. The present report discusses a pregnant patient with Candida albicans sepsis and endophthalmitis as well as candida infection of the oral and genital mucous membranes, after hyperalimentation and broad spectrum antibiotic therapy via a central venous catheter. The patient was treated with 10 mg/kg fluconazole from week 16 of gestation for a total duration of 50 days. Adverse effects did not occur and the rest of the pregnancy proceeded favourably for both the mother and the baby.

Abortion, Septic↗
PubMed1993

Detection of antibodies to Candida albicans germ tube as a possible aid in diagnosing systemic candidiasis in bone marrow transplant patients.

Indirect immunofluorescence assays to detect antibodies to Candida albicans blastospore and germ tube were performed in sera of 29 bone marrow transplant patients. Antibodies to germ tube were present in the sera of six patients, in four of whom a Candida albicans infection was highly probable, while in the other two patients it was not possible to determine the previous course. No healthy blood donors had these antibodies. On the other hand, detection of antibodies to Candida albicans blastospore showed low specificity in the diagnosis of systemic candidiasis. These preliminary findings suggest that the detection of antibodies to Candida albicans germ tube may be an important aid in the diagnosis of systemic candidiasis in bone marrow transplant patients.

Adolescent↗
PubMed1989

Effects of cilofungin (LY121019) on carbohydrate and sterol composition of Candida albicans.

Cilofungin (LY 121019) is a novel analogue of echinocandin B with potent activity against Candida albicans. The effects of cilofungin on the sterol and cell wall carbohydrate composition of Candida albicans were investigated. Exposure of Candida albicans to cilofungin resulted in a 55-60% decrease in ergosterol and a 4-13% decrease in lanosterol content relative to controls. Carbohydrate analysis revealed a 72-79% decrease in glucan content and no significant decrease in mannan or chitin content relative to controls. These studies suggest that cilofungin specifically inhibits glucan synthesis in Candida albicans and thus may be less toxic to mammalian cells. The effects of cilofungin on sterol composition may be non-specific and will require additional investigation.

Candida albicans↗
PubMed1989

Value of the Cand-Tec Candida antigen assay in the diagnosis and therapy of systemic candidiasis in high-risk patients.

A total of 911 sera from 171 patients at risk for systemic candidiasis and 24 sera from 24 non-hospitalized control subjects were analyzed for the presence of candida antigen using a commercially available latex agglutination test (Cand-Tec). Thirty-seven (22%) patients had systemic candidiasis documented by positive blood cultures, deep biopsy culture and histopathology or autopsy. Six patients had transient candidemia, 20 patients had candiduria, 53 patients had mucous membrane colonization, 21 patients were not colonized but received empiric amphotericin B, and 34 patients were not colonized and not treated with amphotericin B. The intraobserver reproducibility was 90% for the exact titer and 100% for a deviation of one dilution. The sensitivity and specificity of the candida antigen test in detection of systemic candidiasis was 95% and 50% (greater than or equal to 1:2), 73% and 72% (greater than or equal to 1:4), and 46% and 80% (greater than or equal to 1:8) respectively. Despite the poor specificity, serial antigen determinations in patients with documented systemic candidiasis demonstrated both an early diagnostic and prognostic role for the candida antigen test. Seventy-one percent of patients whose antigen titer increased during the course of amphotericin B therapy of documented infection died versus only 13% of those whose titer decreased while on therapy (p = 0.01). The candida antigen test has a limited yet potentially useful role in the diagnosis and management of systemic candidiasis in high-risk patients.

Amphotericin B↗
PubMed2002

Candidal urinary tract infections caused by non-albicans Candida species.

The incidence of non-albicans Candida and non-Candida species isolated from the urine of patients admitted to various departments of the Faculty Hospital of the Medical Faculty of Safárik University in Kosice was examined. From a total of 94 samples of analyzed urine 58 strains of C. albicans and 36 strains of yeasts belonging to 6 species of non-albicans Candida and non-Candida spp. were detected: C. parapsilosis (n = 23), C. tropicalis (6), C. krusei (3), C. robusta (2), C. catenulata (1) and Cryptococcus neoformans (1). In relation to the diagnosis, the yeasts were isolated from patients suffering from a kidneys disease, epididymitis, diabetes, neoplastic diseases, urogenital anomalies, obstructive uropathy, cystitis, prostatitis, hemolytic-uremic syndrome, and others.

Candida↗
PubMed1995

T helper cell dichotomy to Candida albicans: implications for pathology, therapy, and vaccine design.

Acquired immunity to Candida albicans is believed to prevent mucosal colonization of adult immunocompetent individuals from progressing to symptomatic infection. Resistance to disease appears to correlate with the detection of delayed-type hypersensitivity responses in vivo and a T helper type 1 (Th1) cytokine secretion profile in vitro. Cellular immunodeficiency, particularly HIV infection, greatly increases the risk of mucosal infection, confirming that CD(4+)-cell-directed immunity is effective locally in controlling infectivity of the yeast. While Th1-type CD4+ cell activation resulting in phagocyte-dependent immunity clearly represents an important mechanism of anticandidal resistance, clinical observations suggest that Th2-type CD4+ cell reactivity may be triggered by Candida antigens in several disease states, including symptomatic infections and immunopathology. This may imply that a Th1-type pattern of reactivity characterizes the saprophytic yeast carriage and resistance to disease by healthy humans, whereas Th2-type responses would be mostly associated with pathology. Moreover, Candida-specific T helper responses, namely humoral and cell-mediated immunity, appear to be reciprocally regulated, as typically occurs in experimental models of parasitic and retroviral infection, where the Th1/Th2 paradigm of acquired immunity has been best characterized. Recent studies, besides providing direct evidence for the occurrence of cross-regulatory Th1 and Th2 responses in mice with candidiasis, emphasize the potential of cytokine/anticytokine therapy for recruiting Candida-specific responses toward protective, Th1-type CD4+ cell reactivity. At the same time, these studies call attention to the possible consequences of C. albicans infection for immunopathology, allergy, and coinfection.

Animals↗
PubMed2004

Occurrence of Candida strains in cases of paronychia.

A total of 43 patients, 11 males and 32 females, with paronychia of the fingernails were examined for the presence of Candida spp. The yeast species isolated were identified using standard laboratory methods, including germ-tube production, morphology on rice agar with Tween 80, and mainly fermentation and assimilation of saccharides. In the male group, two Candida species were detected: C. albicans as the dominant species in 9 patients and C. parapsilosis in 2 cases. Similarly, C. albicans was the prevalent species also in females (n = 17); other Candida species detected were C. parapsilosis (n = 7), C. tropicalis (5) and C. krusei (3). In addition to the genus Candida, the following anaerobic and aerobic microorganisms were isolated from patients of both groups: Fusobacterium spp., Bacteroides spp., Staphylococcus aureus, alpha-hemolytic streptococci, group A beta-hemolytic streptococci, Klebsiella pneumoniae, Neisseria spp. and Pseudomonas aeruginosa.

Adult↗