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PubMed2005

Candida yeast long chain fatty alcohol oxidase is a c-type haemoprotein and plays an important role in long chain fatty acid metabolism.

The industrial yeasts Candida tropicalis or Candida cloacae are able to grow on a variety of long chain alkanes and fatty acids as the sole carbon source. The complete oxidation of these substrates involves two sequential oxidative pathways: omega-oxidation, comprising the P450 alkane oxidase, a flavin-dependent membrane-bound long chain fatty alcohol oxidase [FAO] and a possible separate aldehyde oxidase [F.M. Dickinson, C. Wadforth, Purification and some properties of alcohol oxidase from alkane-grown Candida tropicalis, Biochem. J. 282 (1992) 325-331], and the beta-oxidation pathway, which utilises acylCoA substrates. We recently purified the membrane-bound long chain fatty alcohol oxidase FAO1 and confirmed it is also a c-type haemoprotein. Multiple isoforms may exist for many of these long chain fatty alcohol oxidases and the in vivo requirements for individual genes with respect to specific substrates are still being elucidated. In vitro reconstitution experiments have demonstrated that in Candida maltosa, the cytochrome P450 52A3 gene product can completely oxidise alkanes to dicarboxylic acids [U. Scheller, T. Zimmer, D. Becher, F. Schauer, W. Schunck, Oxygenation Cascade in Conversion of n-Alkanes to, -Dioic Acids Catalyzed by Cytochrome P450 52A3, J. Biol. Chem. 273 (1998) 32528-32534], potentially obviating requirements for a long chain alcohol oxidase. Here, we directly determine in vivo the role of the long chain alcohol oxidase (FAOT) in C. tropicalis, grown on a variety of substrates, followed by gene deletion. The faot double knockout has no detectable faot activity and is incapable of growth on octadecane, but it grows well on oleic acid, palmitic acid and shorter chain alkanes/fatty acids. A spontaneous mutation[s] may have occurred in the faot double gene knockout of C. tropicalis resulting in its inability to grow on oleic acid and hexadecane. The mutations demonstrate that different pathways of octadecane, hexadecane, oleic acid and palmitic acid utilisation exist in C. tropicalis.

Alcohol Oxidoreductases↗
PubMed2006

Discovery of uracil-based histone deacetylase inhibitors able to reduce acquired antifungal resistance and trailing growth in Candida albicans.

Among fungal pathogens such as Candida albicans, acquired drug resistance has not been associated with plasmids or other transferable elements, but it is thought to involve primarily mutations and genetic or epigenetic phenomena. This prompted us to test some histone deacetylase inhibitors (HDACi) from our library, in combination with fluconazole, against C. albicans strains in vitro. Among the tested compounds, the two chloro-containing uracil-hydroxamates 1c and 1d showed a strong reduction of the MIC values on Candida strains that show the trailing growth effect. In this assay, 1c,d were more potent than SAHA, a well-known HDAC inhibitor, in reducing the Candida growth. More interestingly, 1c,d as well as SAHA were able to inhibit the fluconazole-induced resistance induction in Candida cultures.

Antifungal Agents↗
PubMed2004

Phagocytosis of Candida albicans by metastatic and non metastatic human breast cancer cell lines in vitro.

Experiments were carried out to investigate the kinetic characteristics of phagocytosis of candida by metastatic (MCF-7 and ZR-75-1) and nonmetastatic (HCC70) breast cancer cell (BCC) lines. Cancer cells were mixed with candida at a ratio of 1:10 and attachment/phagocytosis were examined using cytospin preparations stained with either Giemsa or tannic acid and May-Grünwald Giemsa and by using flow cytometry. A high attachment of candida to cancer cells (29-39%) was detected as early as 10 min. Following attachment, cancer cells phagocytized yeast. The phagocytic activity of MCF-7 and ZR-75 cells was significantly higher (58-61%) than that of HCC70 cells (26%). A similar trend was observed with respect to the phagocytic index. Phagocytosis of candida by tumor cells was inhibited significantly by both cytochalasin B (50%) and by lowering temperature to 4 degrees C (66%). Phagocytosis was not associated with oxidative burst in any cell lines used. In conclusion, metastatic breast cancer cells possess phagocytic activity which may explain their invasive property.

Breast Neoplasms↗
PubMed2004

In vitro activities of voriconazole, posaconazole, and fluconazole against 4,169 clinical isolates of Candida spp. and Cryptococcus neoformans collected during 2001 and 2002 in the ARTEMIS global antifungal surveillance program.

We examined the in vitro activities of voriconazole, posaconazole, and fluconazole against 3,932 isolates of Candida spp. and 237 isolates of Cryptococcus neoformans obtained from over 100 medical centers worldwide during 2001 and 2002. The MICs of the antifungal drugs were determined by broth microdilution tests performed according to the National Committee for Clinical Laboratory Standards (NCCLS) methods using RPMI 1640 as the test medium. Voriconazole and posaconazole were very active against Candida spp. (97-98% susceptible at MICs < or =1 microg/ml) and C. neoformans (98-100% susceptible at MICs < or =1 microg/mL). C. albicans (MIC90, 0.015-0.03 microg/ml) was the most susceptible species of Candida to both agents and C. glabrata (MIC90, 1-2 microg/mL) was the least susceptible. Both voriconazole and posaconazole were more active than fluconazole against all Candida spp. and C. neoformans. These results provide further evidence for the increased spectrum and potency of the new triazoles against a large and geographically diverse collection of opportunistic fungal pathogens.

Candida↗
PubMed2004

Tolerance to amphotericin B in clinical isolates of Candida tropicalis.

A broth microdilution method was used for testing amphotericin B against 33 clinical isolates of Candida tropicalis. All isolates were in vitro susceptible to the polyene (MIC [minimal inhibitory concentration] < or = 1.0 microg/mL). However, when the isolates were cultured in a medium containing amphotericin B at a concentration of 1.5 microg/mL, a wide interstrain variation of growth rate was observed. Five isolates (15%) proved to be highly tolerant to the drug and grew at a frequency ranging from 1 x 10(-1) to 2 x 10(-2). Twenty-three isolates (70%) grew at a frequency ranging from 1 x 10(-5) to 1 x 10(-8). The remaining five isolates (15%) failed to grow in drug-containing medium. In general, this growth variation was not associated with amphotericin B MICs displayed by the single isolates. In addition, the strains grown in drug-containing medium did not represent amphotericin B-resistant mutants, as shown by the maintenance of MICs similar to those of their respective parent isolates. Killing experiments conducted in selected isolates confirmed a variation of fungicidal activity of amphotericin B. To see whether this phenomenon was associated with a variation of amphotericin B response in vivo, we established an experimental model of systemic murine candidiasis in CD1 mice by intravenous injection of cells belonging to Candida tropicalis 3147 (growth rate at a frequency of 1 x 10(-1) in amphotericin B medium) and Candida tropicalis 4055 (no growth). Low (0.3 mg/kg/day) and high (1 mg/kg/day) doses of amphotericin B were both effective at reducing the fungal burdens in the kidneys of mice infected with either strain (p, 0.01 to 0.02). However, whereas the burden of mice infected with isolate 3147 and treated with the polyene at 0.3 mg/kg/day was reduced by 1.2 +/- 0.25 (mean +/- standard deviation) log10 cfu/g compared to untreated mice, the same dosing regimen yielded a burden reduction of 2.6 +/- 0.07 log10 cfu/g in mice infected with isolate 4055 (p < 0.001). Similarly, amphotericin B at 1 mg/kg/day yielded a burden reduction of 1.8 +/- 0.20 vs. 2.5 +/- 0.30 log10 cfu/g in mice infected with isolates 3147 and 4055, respectively (p < 0.001). Our data revealed a variable pattern of tolerance to amphotericin B among isolates of Candida tropicalis and showed that this phenomenon might influence the rate of organ clearance during therapy.

Amphotericin B↗
PubMed2004

Anti-retroviral therapy with protease inhibitors decreases virulence enzyme expression in vivo by Candida albicans without selection of avirulent fungus strains or decreasing their anti-mycotic susceptibility.

Highly active anti-retroviral therapies (HAART) with human immunodeficiency virus (HIV) protease inhibitors (PIs) or nonnucleoside reverse-transcriptase inhibitors (NNRTI) were compared for their effect on prevalence, aspartyl proteinase (Sap) production and the biotypes and anti-mycotic sequential susceptibility of Candida spp. isolates from the oral cavity in a longitudinal prospective study. HAART-PI, but not HAART-NNRTI strongly inhibited Sap expression in the oral cavity without exerting any consistent effect on the role of Candida spp. isolation or selection of low virulence or anti-mycotic resistant fungus biotype. More importantly, the sequential isolates of Candida albicans from HAART-PI, but not those from suspended HAART-NNRTI, showed an increased Sap production in vitro. While further demonstrating that HIV-PI inhibit Sap expressions, our results do not support the view that the mentioned inhibition could eliminate Candida or its selection of the oral cavity.

AIDS-Related Opportunistic Infections↗
PubMed2006

Antifungal resistance in Candida spp. isolated in Italy between 2002 and 2005 from children and adults.

The activity of amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole was tested in vitro against 618 clinical Candida spp. isolates, using the broth microdilution or the disk diffusion method (voriconazole). Amphotericin B and voriconazole were the most potent antifungal agents assayed (100% of susceptible strains). Resistance to fluconazole and itraconazole was detected in three (0.7%) and 11 (2.7%) isolates of Candida albicans and in four (3.7%) isolates of Candida glabrata. Flucytosine intermediate, resistant strains, or both, were observed in C. albicans (0.3% and 0.7%), C. glabrata (2.8% intermediate) and C. tropicalis (15.2% and 15.2%). C. krusei was the least susceptible species to azoles. No statistically significant differences in the rates of resistant isolates depending on site of infection and age of the patient were observed, with the exception of C. albicans and itraconazole (higher percentage of resistance in children). At present, acquired antifungal resistance represents an uncommon finding in most Candida spp. circulating in Northern Italy.

Adult↗
PubMed2026

Machine learning-based drug susceptibility prediction from Candida genomic data.

OBJECTIVES: Invasive Candida infection is an increasing clinical concern, with antifungal resistance rising across multiple species. However, rapid and accurate antifungal susceptibility testing (AFST) remains limited in routine practice. The study evaluated species distribution and antifungal susceptibility of invasive Candida isolates in China and assessed the feasibility of combining whole-genome sequencing (WGS) with machine learning to predict minimum inhibitory concentrations (MICs). METHODS: Consecutive non-repetitive isolates were collected from 20 hospitals in 13 provinces during 2022-2023. MICs of nine antifungal agents were determined by broth microdilution, and WGS was performed for species accounting for >5% of the total isolates. Genomic 11-mer features were extracted and used to train random forest (RF), support vector machine (SVM), and extreme gradient boosting (XGBoost) models, followed by optimization of the best-performing algorithm. RESULTS: A total of 337 isolates were obtained from blood (n = 232) and sterile body fluids (n = 105), comprising C. albicans (n = 103), C. tropicalis (n = 71), C. parapsilosis (n = 67), and C. glabrata (n = 63). Non-albicans Candida showed higher azole and echinocandin resistance, with C. tropicalis notably resistant to azoles and C. glabrata to echinocandins. Among the three models, RF demonstrated the best performance on 304 sequenced isolates. The optimized RF model was evaluated by the receiver operating characteristic (ROC) curve analysis and achieved an average area under the ROC curve (AUC) of 0.979 (95% CI: 0.974-0.984), essential agreement over 90.1%, and categorical agreement over 93.2% across species. CONCLUSIONS: These findings underscore the clinical challenge posed by non-albicans Candida resistance, and indicate that WGS-based MIC prediction may offer a highly accurate reference for earlier antifungal therapy.

Antifungal Agents↗
PubMed2004

Candida albicans osteomyelitis: case report and literature review.

OBJECTIVE: Osteomyelitis due to Candida species is an unusual but recognized entity. However, with the increasing occurrence of factors predisposing to candidemia and invasive candidiasis, candidal osteomyelitis is being diagnosed more frequently. An unusual case of Candida albicans osteomyelitis is reported here, along with a review of the published literature on previously reported cases of this disease. METHODS: Report of the case and literature review. RESULTS: In this case, Candida albicans was isolated from the talus; however, the diagnosis was made after several cultures were performed. Only one other case of Candida osteomyelitis located in foot bones was found in the review. CONCLUSIONS: It is thought that this is the first case reported in Venezuela, and only the second in international literature.

Aged↗
PubMed2006

Influence of leflunomide on gastrointestinal Candida albicans infection induced in naive and arthritic newborn mice.

Mucosal Candida albicans infection in immunocompromised individuals being treated with recently advanced drugs can progress to systemic disease. One such medication applied in patients with rheumatoid arthritis is leflunomide. The object of the present study was to investigate the effect of leflunomide on a model of gastrointestinal (g.i.) C. albicans infection induced in naïve or arthritic mice and on the host resistance to systemic re-infection. Newborn mice were orally inoculated with 1 x 10(5) colony forming units (CFU) of C. albicans and at age of 5 weeks they were treated with 5 mg/kg or 20 mg/kg of leflunomide for 10 consecutive days. Both doses elevated the yeast colonization of the stomach, without the dissemination into the internal organs. This was in parallel with the enhanced delayed type hypersensitivity (DTH) reaction to the yeast. Contrary to that, leflunomide caused a shift to Th2 reactivity by prevalence of IL-4 to IFN-gamma and a suppression of anti-Candida antibody synthesis by a higher dose. It might be supposed that infection increased autoimmune response in arthritic mice, according to stimulated DTH reaction to collagen. The administration of leflunomide during the simultaneous development of infection and arthritis diminished anti-Candida and anti-collagen antibody synthesis compared to untreated infected arthritic mice. The improved survival of arthritic infected animals against severe systemic re-infection was not changed after administration of leflunomide to re-infected arthritic mice. We can conclude that although leflunomide influenced cytokine secretion and suppressed anti-Candida antibody production it neither provokes a progression from gastrointestinal to systemic C. albicans infection nor increases the susceptibility to severe C. albicans re-infection of arthritic mice.

Animals↗
PubMed2003

Long-term epidemiological survey of Candida species: comparison of isolates found in an intensive care unit and in conventional wards.

The aim of this study was to compare the distribution of Candida species in patients hospitalized in an intensive care unit (ICU) and in conventional wards. A retrospective analysis was performed covering an 18-year period in a 700-bed teaching hospital. Various body sites were investigated in all patients admitted during the study and isolates were identified by microscopic and macroscopic morphology, and by commercially available kits. The susceptibility of strains to amphotericin B and flucytosine was assessed by the ATB-fungus system, itraconazole and fluconazole by Etest. No difference was observed between the distribution of Candida species in ICU and in conventional wards. Candida albicans represented about 70% of isolates and Candida glabrata was the second most common species involved in infection or colonization. The small number of C. glabrata resistant to fluconazole suggested this antifungal agent as suitable empirical treatment for non-immunocompromized patients in whom a fungal infection was suspected.

Antifungal Agents↗
PubMed2005

Twice weekly fluconazole prophylaxis for prevention of invasive Candida infection in high-risk infants of <1000 grams birth weight.

OBJECTIVES: We tested the hypothesis that twice weekly prophylactic dosing of fluconazole prevents invasive candidiasis without promoting resistant Candida species in high-risk, preterm infants. STUDY DESIGN: We compared our previous dosing schedule (Group A) to a less frequent dosing schedule of twice a week (Group B) of fluconazole prophylaxis for up to 6 weeks in a prospective, randomized, double-blind clinical trial in preterm infants weighing <1000 grams at birth and with an endotracheal tube and/or central vascular catheter over a 24-month period. Weekly surveillance cultures were obtained on study patients. RESULTS: Candida colonization was documented in 5 (12%) of 41 Group A and in 4 (10%) of 40 Group B infants. Candida sepsis developed in two (5%) of Group A and one (3%) of Group B infants (risk difference, -0.02; 95% confidence interval, -0.14-0.10; P=.68). All fungal isolates remained sensitive to fluconazole, and no drug side effects were documented. CONCLUSIONS: Twice weekly dosing of prophylactic fluconazole can decrease Candida colonization and invasive infection, cost, and patient exposure in high-risk, preterm infants weighing <1000 grams at birth. We speculate that lower and less frequent dosing may delay or prevent the emergence of antifungal resistance.

Antifungal Agents↗
PubMed2002

Genotyping and antifungal susceptibility of human subgingival Candida albicans isolates.

Subgingival colonization by Candida albicans has been described in human immunodeficiency virus (HIV)-infected individuals, but subgingival isolates have scarcely been characterized, particularly with respect to genotype and antifungal susceptibility. A series of 29 subgingival strains of C. albicans isolated from nine HIV-infected individuals was typed by electrophoretic karyotyping and tested for susceptibility to fluconazole, itraconazole, the new investigational triazole posaconazole and amphotericin B. DNA typing showed genetic heterogeneity within subgingival isolates, as almost every individual harbored his/her own specific isolate. Genetic identity was usually demonstrated within oral and subgingival isolates simultaneously collected from the same individual, but a number of DNA types were found to be unique to subgingival strains. These findings suggest that colonization is not just the result of Candida spreading from oral surfaces, and that subgingivally adapted strains could be involved. All isolates were susceptible to all the triazole drugs tested and amphotericin B. Additional studies on subgingival Candida colonization and further characterization of subgingival isolates are now required to clarify the role of Candida as opportunistic periodontal pathogen.

Adult↗
PubMed2003

Structural characterization of (1-->3)-beta-D-glucans isolated from blastospore and hyphal forms of Candida albicans.

Glucans are (1-->3)-beta-linked linear and branched polymers containing anhydroglucose repeat units. They comprise a major portion of the cell wall of saprophytic and pathogenic fungi. Glucans activate a wide range of innate immune responses. They are also released from the fungal cell wall as exopolymers into the blood of patients with fungal infections. Extensive studies have been done on glucans isolated from saprophytic fungi, such as Saccharomyces cerevisiae; however, much less is known about the glucans produced by the polymorphic fungal pathogen Candida albicans. We have undertaken an extensive structural characterization and comparison of glucans isolated from C. albicans blastospores and hyphae using high-resolution, solution-state proton nuclear magnetic resonance spectroscopy (NMR). In addition, we developed a simple and straightforward method for the production of Candida hyphae that resulted in gram quantities of hyphal mass. Also, we compared and contrasted the Candida glucans isolated by two different protocols with those isolated from S. cerevisiae. Isolation protocols provide high purity glucans with source-based structural differences. Structural details provided by this NMR analysis included the degree of polymerization, molecular weight, degree and type of branching, and structural composition. We observed that Candida glucans, derived from blastospores or hyphae, are different compared to those isolated from S. cerevisiae with regard to side-chain branching along the backbone and at the reducing terminus. These structural details are an important prerequisite for biomedical studies on the interaction of isolated fungal cell wall glucans with the innate immune system.

Candida albicans↗
PubMed1998

Intact autocrine activation and cytokine production by PMNs from injured adults with elevated Candida antigen titres.

Injured patients with Candida antigen titres have increased mortality due to sepsis. Polymorphonuclear leucocytes (PMNs) from injured patients with elevated Candida antigen titres demonstrate impaired function against Candida albicans growth when compared with PMNs from injury matched controls. To determine if PMN dysfunction is global, PMNs from patients with positive Candida antigen titres were evaluated for their ability to activate the anticandidal function of normal PMNs (autocrine activation) and to produce tumour necrosis factor (TNF) and interleukin 8 (IL8), known activators of PMN anticandidal function, this study demonstrates that the PMN dysfunction is not global, as PMN cytokine production and autocrine activation remain intact.

Adult↗
PubMed1996

Denture stomatitis: quantification of interleukin-2 production by mononuclear blood cells cultured with Candida albicans.

Denture stomatitis is usually associated with the presence of yeast, particularly Candida albicans, and several bacteria. In this study mononuclear blood cells were grown in the presence of Candida albicans from a single colony, and interleukin-2 production induced in T lymphocytes was measured. Blood cells were from a population of patients with denture stomatitis and a control group of denture wearers without stomatitis. Induction of interleukin-2 production was correlated with factors that condition denture stomatitis, namely, isolation of Candida albicans in selective medium, age of the denture, and diabetes. Concentrations of interleukin-2 in supernatant and serum were also compared. Significant differences in interleukin-2 production were found between patients with denture stomatitis and controls. Statistical analysis demonstrated a significant association between isolation of Candida albicans and elevated interleukin-2 production in cultures from patients with and without denture stomatitis.

Adult↗
PubMed1989

Clinical significance of Candida isolated from peritoneum in surgical patients.

Over a 2-year period, all surgical patients from whom Candida was isolated from intra-abdominal specimens were evaluated. All but 1 of the 49 evaluable patients had either a spontaneous perforation (57%) or a surgical opening of the gastrointestinal tract (41%). Candida caused infection in 19 patients (39%), of whom 7 had an intra-abdominal abscess and 12 peritonitis. In the other 30 patients (61%), there were no signs of infection and specific surgical or medical treatment was not required. Candida was more likely to cause infection when isolated in patients having surgery for acute pancreatitis than in those with either gastrointestinal perforations or other surgical conditions. The development of a clinical infection was significantly associated with a high initial or increasing amount of Candida in the semiquantitative culture. Surgery alone failed in 16 of 19 patients (84%), of whom 7 died and 9 recovered after combined antifungal and surgical treatment. The overall mortality and the mortality related to infections were significantly higher in the patients with intraabdominal candidal infections than in those without such infections.

Abscess↗
PubMed1996

Corneal anesthetic abuse and Candida keratitis.

PURPOSE: Topical corneal anesthetic abuse is a self-inflicted injury, causing profound corneal morbidity. Superimposed infection is an important complicating factor. The authors report four patients with confirmed topical anesthetic abuse of the cornea, in whom Candida keratitis developed. METHODS: A retrospective review of the medical records of four patients with confirmed topical corneal anesthetic abuse and fungal keratitis. RESULTS: A 21-year-old woman, two 28-year-old women, and a 35-year-old man were included in the study. All these patients sustained a corneal injury, prompting the chronic use of topical anesthetics (0.5% proparacaine hydrochloride in 3 patients, and 0.5% tetracaine hydrochloride and 0.4% benoxinate hydrochloride in the other). Corneal findings included epithelial defects in all patients, focal infiltrate in one patient, and ring-shaped stromal infiltrate in three patients. Topical anesthetic was discontinued, all patients initially were treated empirically with antibacterial agents, and three patients received topical corticosteroids. Subsequent corneal cultures grew Candida spp, Candida albicans specifically in three patients, and local and systemic antifungal therapy was started. Corneas in two patients re-epithelialized; a conjunctival flap was performed on another patient with a descemetocele; and the remaining patient was lost to follow-up, although repeat fungal cultures yielded no growth. CONCLUSIONS: Corneal superinfection with Candida may occur during topical anesthetic abuse. Therapy includes discontinuation of the anesthetic and institution of antifungal therapy.

Administration, Topical↗