Candida albicans

FEMS Microbiol Lett, 1997 Jul 15, 152(2), 235 - 42
Amended structure of side chains in a cell wall mannan from Candida albicans serotype A strain grown in yeast extract-Sabouraud liquid medium under acidic conditions: detection of the branched side chains corresponding to antigenic factor 4; Kobayashi H et al.; In a previous study, we reported the excess production of alpha-1,3-linked mannose residues with the complete disappearance of beta-1,2-linked mannose residues in cell wall mannans of Candida albicans serotype A strain cells, which were grown in yeast extract-Sabouraud liquid medium at pH 2.0 . In the present study, we examined the immunochemical reactivity of the same mannan of NIH A-207 with an enzyme-linked immunosorbent assay (ELISA) using several antisera to antigenic factors of the genus Candida (FAbs) and the structure of the mannan by two-dimensional homonuclear Hartmann-Hahn analysis . The ELISA showed that the mannan reacts to FAb 4 but not to FAbs 13b and 34, which are reported to be antibody factors against linear side chains containing an alpha-1,3-linked mannose residue . In the Hartmann-Hahn analysis, we found two branched side chains, Man alpha 1-2Man alpha 1-3{Man alpha 1-6}Man alpha 1-(2Man alpha 1-)(2)2Man and Man alpha 1-3{Man alpha 1-6}Man alpha 1-(2Man alpha 1-)(2)2Man, instead of the previously reported linear side chains . The branched side chains are oversynthesized under acidic conditions.

Science, 1997 Jul 4, 277(5322), 105 - 9
Control of filament formation in Candida albicans by the transcriptional repressor TUP1; Braun BR et al.; The pathogenic yeast Candida albicans regulates its cellular morphology in response to environmental conditions . Ellipsoidal, single cells (blastospores) predominate in rich media, whereas filaments composed of elongated cells that are attached end-to-end form in response to starvation, serum, and other conditions . The TUP1 gene, which encodes a general transcriptional repressor in Saccharomyces cerevisiae, was isolated from C . albicans and disrupted . The resulting tup1 mutant strain of C . albicans grew exclusively as filaments under all conditions tested . TUP1 was epistatic to the transcriptional activator CPH1, previously found to promote filamentous growth . The results suggest a model where TUP1 represses genes responsible for initiating filamentous growth and this repression is lifted under inducing environmental conditions.

J Biol Chem, 1997 Jul 4, 272(27), 16822 - 8
Characterization of beta-1,2-mannosyltransferase in Candida guilliermondii and its utilization in the synthesis of novel oligosaccharides; Suzuki A et al.; A particulate insoluble enzyme fraction containing mannosyltransferases from Candida guilliermondii IFO 10279 strain cells was obtained as the residue after extracting a 105,000 x g pellet of cell homogenate with 1% Triton X-100 . Incubation of this fraction with a mannopentaose, Manalpha1-->3(Manalpha1-->6)Manalpha1-->2Manalpha1+ ++-->2Man, in the presence of GDP-mannose and Mn2+ ion at pH 6.0 gave a third type of beta-1,2 linkage-containing mannohexaose, Manbeta1-->2Manalpha1-->3(Manalpha1-->6)Manalpha1++ +-->2Manalpha1-->2Man , the structure of which was identified by means of a sequential NMR assignment . The results of a substrate specificity study indicated that the beta-1,2-mannosyltransferase requires a mannobiosyl unit, Manalpha1--> 3Manalpha1-->, at the nonreducing terminal site . We synthesized novel oligosaccharides using substrates possessing a nonreducing terminal alpha-1,3-linked mannose unit prepared from various yeast mannans . Further incubation of the enzymatically synthesized oligosaccharide with the enzyme fraction gave the following structure, Manbeta1-->2Manbeta1-->2Manalpha1-->3(Manalpha1- ->6)Manalpha1--> 2Manalpha1-->2Man, which has been found to correspond to antigenic factor 9 . Incubation of Candida albicans serotype B mannan with the enzyme fraction gave significantly transformed mannan, which contains the third type of beta-1,2-linked mannose units.

Maturitas, 1997 Jul, 27(3), 253 - 60
The relationship of bacterial vaginosis, Candida and Trichomonas infection to symptomatic vaginitis in postmenopausal women attending a vaginitis clinic; Spinillo A et al.; OBJECTIVE: To estimate the prevalence of bacterial vaginosis, Candida albicans, and Trichomonas vaginalis infections in a population of postmenopausal women with symptoms of vaginitis seen at a vaginitis clinic either as self-referred or clinician referred patients . METHODS: A cross-sectional study of 148 postmenopausal women (cases) and 1564 controls of reproductive age attending a vaginitis clinic . C . albicans and T . vaginalis infections were diagnosed by culture techniques . Bacterial vaginosis was diagnosed on the basis of clinical findings . RESULTS: Fifty-six (37.8%) postmenopausal women and 834 (53.3%) controls were diagnosed with T . vaginalis or C . albicans infection, or bacterial vaginosis, or mixed infection (odds ratio (OR) 0.53, 95% confidence interval (CI) 0.37-0.75) . C . albicans and T . vaginalis infection were diagnosed in 34.1% (534/1564) and 1.92% (30/1564) of women of childbearing age and in 13.5% (20/148) and 10.8% of postmenopausal women, respectively . (P < 0.05 for both comparisons) . The prevalence of bacterial vaginosis was similar between the two groups (14/148 in postmenopausal patients and 210/1564 in controls of reproductive age; P = 0.22) . CONCLUSIONS: Among postmenopausal women attending a vaginitis clinic, a defined diagnosis of bacterial vaginosis, C . albicans or T . vaginalis infection can be made in about one third of such patients . Concerning the two thirds of symptomatic women lacking such a microbiologic diagnosis, alternative causes (e.g., estrogen deficiency, nonanaerobic bacterial infections, local irritants or allergenes, and dermatologic conditions) need to be considered.

Mol Microbiol, 1997 Jul, 25(2), 229 - 36
Beta, a novel repetitive DNA element associated with tRNA genes in the pathogenic yeast Candida albicans; Perreau VM et al.; We have identified a novel 399 bp repetitive DNA element (which we designate beta) 9bp upstream of a seryl-tRNA(CAG) gene in the genome of Candida albicans . There are two copies of the seryl-tRNA(CAG) gene, one on each homologue of chromosome VI, and the beta element is found upstream of one copy of the gene in C . albicans strain 2005E . The beta element is not present upstream of either copy of the seryl-tRNA(CAG) gene in eight other laboratory strains of C . albicans tested, but was detected in this location in several fresh clinical isolates . Southern blot analysis indicated that there are approximately eight copies of the beta element per diploid C . albicans genome and that it is a mobile element, being present on at least two different chromosomes . Three unique genomic DNA clones containing the beta element were isolated from strain 2005E; in each case, a different tRNA gene was found immediately adjacent to the beta element . Three new tRNA genes from C . albicans have thus been identified: tRNA(Asp), tRNA(Ala) and tRNA(Ile) . The beta element shows no significant sequence homology to other known prokaryotic or eukaryotic repetitive elements, although an 8 bp repeat at the 3' end of the element is identical to that of the Ty3 retrotransposable element of Saccharomyces cerevisiae . We propose that the beta element is a solo long terminal repeat (LTR) sequence of a Ty3/gypsy-like transposable element in C . albicans that is closely associated with tRNA genes.

Toxicol Pathol, 1997 Jul-Aug, 25(4), 351 - 62
Single-organism model of host defense against infection: a novel immunotoxicologic approach to evaluate immunomodulatory drugs; Herzyk DJ et al.; The immunotoxicologic effects of drugs on host defense have been studied widely using various animal models of infection . Here we describe a new approach to testing host defense by using a single organism (Candida albicans) in CBA/J mice . The model is configured to test 3 effector systems via different routes of inoculation to stimulate different effector arms of the immune response . Nonspecific immunity was evaluated by C . albicans colony-forming unit (CFU) count from the spleen at 2 hr (uptake) and > or = 22 hr (clearance) following intravenous inoculation . Cell-mediated immunity was assessed by CFU count from an intramuscular injection site 6 days postinoculation . Humoral immunity was assessed by anti-Candida antibody titer, following multiple subcutaneous immunizations with C . albicans . Finally, overall immunity was evaluated following intravenous injection using survival as the endpoint . Histopathological, immunohistochemical, and electron microscopic evaluation of selected tissues revealed the involvement of the expected cell types in the different effector systems . Several immunomodulatory drugs--dexamethasone, cyclosporine, liposomal muramyltripeptide phosphatidylethanolamine, and SK&F 105685--were evaluated in the C . albicans model . Dexamethasone impaired host defense against C . albicans by suppressing all endpoints measured . Similarly, cyclosporine showed broad immunosuppressive activity, with the exception of yeast uptake from the spleen . In contrast, muramyl tripeptide-phosphatidylethanolamine enhanced all but cell-mediated immunity to C . albicans . SK&F 105685 displayed both stimulatory and inhibitory effects on immune responses to the infection . Our studies demonstrate that a single organism-based approach can be a useful method for evaluating the immunological hazards of drugs on host resistance to infection.

Br J Dermatol, 1997 Jul, 137(1), 76 - 80
Adherence of Candida albicans strains isolated from AIDS patients . Comparison with pathogenic yeasts isolated from patients without HIV infection; Pereiro M Jr et al.; The adherence of yeasts to oral mucous cells is one of the main characteristics of the pathogenicity of this fungus . We studied adherence by means of a radiometric test to improve the method . We compared a sample of 40 strains of Candida albicans isolated from the buccal mucosa of HIV-infected patients with 40 strains isolated from non-HIV patients . We found that buccally isolated C . albicans strains from patients in the initial stages of AIDS adhered to oral mucous cells less than the buccally isolated C . albicans strains from subjects without HIV infection . Adherence among the strains of HIV patients increased with the disease stage until it exceeded that of the normal subjects in proportion to the decrease in the CD4/CD8 ratio . The selection of resistant strains by the preliminary antifungal treatments gave us a partial explanation for this increase . Further research should be carried out to compare these results with those obtained from atypical strains and species with high pathogenic potential, such as Candida dubliniensis, which is frequently isolated from advanced AIDS, in order to prevent systemic infections in these patients.

FEMS Immunol Med Microbiol, 1997 Jul, 18(3), 147 - 52
Expression and quantification of the iC3b-binding protein in different Candida albicans strains and their morphological stages; Bujdakova H et al.; Expression and quantification of the iC3b-binding protein in yeast, germ-tube, and mycelial forms of several Candida albicans strains were studied . Ten isolates were obtained from patients with recurrent vaginal candidosis . The germ-tubes generated at 37 degrees C and the mycelial forms of all strains grown at 30 degrees C, as well as most of the mycelial forms grown at 37 degrees C, were able to bind complement-coated sheep erythrocytes (EAiC3b) . ELISA results revealed that a decrease in the binding of EAiC3b by the mycelial form of strains CBS 5982 and K10 correlated with a decrease of the expression of the iC3b-binding protein, detected by cross-reaction with the monoclonal antibody OKM1, recognising the alpha chain of human CR3 . Expression of the iC3b-binding protein in other strains, binding EAiC3b, was higher in the mycelial form or very similar to that of germ-tubes . The dependence of the expression of the iC3b-binding protein on the morphological stages of individual C . albicans isolates suggests a possible association with the virulence of these strains.

J Antimicrob Chemother, 1997 Jul, 40(1), 19 - 25
In-vitro studies of two 5-nitroimidazole derivatives; Castelli M et al.; This paper reports the findings obtained using two new compounds belonging to the 5-nitroimidazole family: sulphuridazole (V1) and sulphonidazole (V2) . We first assessed their antimicrobial activity on Clostridia spp . and then extended the study to Gram-positive and Gram-negative aerobic microorganisms and to Candida albicans . Their MICs were compared with those of metronidazole . The findings show that the antibacterial and antimycotic activity of sulphonidazole is greater than that of sulphuridazole, while metronidazole is not active against any aerobic organism . It also emerges that the NO2 group is indispensable for all the microorganisms assayed and that sulphuridazole and sulphonidazole are the first two 5-nitroimidazoles active against C . albicans . The redox potentials of the 5-nitroimidozoles studied suggest that their action mechanism is mainly based on redox processes.

Gene, 1997 Jul 1, 193(1), 115 - 8
Nucleotide sequence of the gene coding for SEC14p in Candida (torulopsis) glabrata; Dundon W et al.; A gene coding for SEC14p from Candida glabrata has been cloned and characterized . Nucleotide (nt) sequence analysis reveals an open reading frame of 909 bp and predicts the synthesis of a polypeptide of 302 amino acid (aa) residues . Comparison of nt and aa sequences shows that the gene exhibits a much higher homology to the Saccharomyces cerevisiae (72% and 87%, respectively) than to the Candida albicans (55% and 65%, respectively) SEC14 gene.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1997 Jul, 84(1), 68 - 73
Growth patterns of Candida albicans in relation to radicular dentin; Sen BH et al.; Candida albicans is the most common fungal pathogen isolated from the oral cavity . The role of this organism as an endodontic pathogen is poorly understood . OBJECTIVES: The aim of this study was to observe the interaction of C . albicans with root canal walls and the growth patterns of this microorganism in relation to radicular dentin . STUDY DESIGN: Fifteen root sections were infected with C . albicans grown in calf serum and incubated for various periods . The sections were fixed in glutaraldehyde, split into two halves, and evaluated by scanning electron microscopy . RESULTS: Blastospores and hyphal structures were observed on the root canal walls of all specimens . Filamentous hyphal form was dominant in 5-day specimens . Most of the hyphae and blastospores showed penetration into dentinal tubules . The body of germinating mother cells and hyphae demonstrated collapsed cell walls as a result of vacuole formation . CONCLUSIONS: With this invasive affinity to dentinal structures, C . albicans may be considered a dentinophilic microorganism.

Yeast, 1997 Jul, 13(9), 871 - 80
Isolation and characterization of the Candida albicans PFY1 gene for profilin; Ostrander DB et al.; We have isolated the Candida albicans gene for profilin, PFY1 . Degenerate oligonucleotide primers based on regions of high homology were utilized to obtain a polymerase chain reaction-amplified copy of the gene . This was then used as a probe to isolate the gene from a C . albicans genomic library . Our studies indicate that the full-length gene is unstable in Escherichia coli . Several clones were sequenced, and the predicted amino acid sequence demonstrated homology with profilin proteins from other organisms, most notably Saccharomyces cerevisiae . Northern analysis revealed that the gene is expressed in C . albicans . Attempts to express the gene in S . cerevisiae cells were unsuccessful until the C . albicans promoter was replaced with an S . cerevisiae promoter . Functional complementation of the gene was demonstrated in S . cerevisiae profilin-requiring cells . Antibodies raised to isolated C . albicans profilin protein recognized a protein of the predicted molecular weight when the gene was expressed in S . cerevisiae cells.

J Oral Pathol Med, 1997 Jul, 26(6), 290 - 3
Exfoliative cheilitis (EC) in AIDS: association with Candida infection; Reichart PA et al.; Forty-seven of 165 patients with AIDS (28.5%) showed exfoliative cheilitis (EC), predominantly of the lower lip (n = 37) . Histologically, hyphae were revealed in 23 of 47 cases (49%) . In 14 of 23 specimens the histological and microbiological findings were in accordance . Smears of the vermilion border revealed Candida albicans in half of the cases (51%); however, combinations with C . krusei, C . tropicalis and C . glabrata were also seen . Twenty of 35 patients given fluconazole either prophylactically or therapeutically showed clinical signs of oral candidiasis . Frequent moistening of the lips may result in infection of the vermilion border with Candida species; consequent desiccation of the lips will lead to scale formation and exfoliation . Smears of the vermilion border of the lower lip of 20 controls with AIDS were positive in four cases . Twenty HIV-negative controls without EC showed negative microbiological results for Candida species . Exfoliative cheilitis may be associated with Candida infection in some cases and may be considered another variant of candidiasis in AIDS patients.

J Leukoc Biol, 1997 Jul, 62(1), 60 - 6
Possible participation of polymorphonuclear cells stimulated by microbial immunomodulators in the dysregulated cytokine patterns of AIDS patients; Cassone A et al.; Macrophages and polymorphonuclear cells (PMN) play a major role as cells primarily responsive to microbial biological response modifiers (BRM) . Although much attention has been given to macrophages, PMN have been relatively underinvestigated . We have recently studied the responses of PMN from HIV- and HIV+ subjects after stimulation with a powerful immunomodulatory fraction from the cell wall of Candida albicans (MP-F2) and compared this to bacterial lipopolysaccharide (LPS) . Both cytokine patterns and PMN anticandidal activity were investigated . MP-F2, like LPS, was an active inducer of interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-alpha), IL-6, and IL-1 beta production by PMN and monocytes from all subjects . IL-12 was also produced by MP-F2-stimulated PMN in the presence of interferon-gamma (IFN-gamma) . PMN from HIV+ subjects showed increased in vitro expression of TNF-alpha and IL-6 genes as determined by semiquantitative reverse transcriptase-polymerase chain reaction . In all subjects, cytokine gene expression was strongly stimulated by MP-F2 or LPS and inhibited by IL-10 . Production of IL-6 and TNF-alpha protein (measured by ELISA) was higher in PMN from HIV+ subjects in at least one of the conditions tested (unstimulated or stimulated by LPS or MP-F2) . However, the amount of the C-X-C chemokine IL-8 was equal in PMN from HIV- and HIV+ subjects . PMN from HIV+ subjects were at least as active in inhibiting candide growth as PMN from HIV- controls . In both groups PMN were equally stimulated by MP-F2 and LPS . Only in severely neutropenic subjects was there some reduction in the anticandidal activity but not in cytokine responses . When appropriately stimulated by microbial BRM, PMN are active producers of pro-inflammatory and immunomodulatory cytokines . This production is not only totally preserved in HIV+ subjects but may be higher than in PMN from HIV- subjects and may be coupled with an efficient anticandida activity . We suggest that during common bacterial or fungal infections PMN may contribute to the dysregulated production of inflammatory cytokines in AIDS patients.

Clin Diagn Lab Immunol, 1997 Jul, 4(4), 447 - 51
Perioperative variation in phagocytic activity against Candida albicans measured by a flow-cytometric assay in cardiovascular-surgery patients; Tran TL et al.; Candidiasis is an opportunistic fungal infection that frequently occurs following modifications of host defenses . Major surgery can be responsible for such alterations, and therefore it increases the risk of fungal infection . The purpose of this study was to evaluate the perioperative impairment of leukocyte function in patients after cardiovascular surgery by measuring the phagocytic activity against Candida albicans by a flow-cytometric method . The average postsurgical decrease in phagocytosis in our patients was 11.4% . By univariate analysis, three factors, all related to antibiotic therapy, were significantly associated with an important decrease in phagocytosis; the use of antimicrobial therapy before surgery, the number of different antibiotics taken, and the length of antibiotic treatment . The results of our study showed that the use of antibiotics in patients undergoing cardiovascular surgery alters the normal phagocytic activity of the host immune system against C . albicans and that flow cytometry is a rapid and simple technique that helps in early identification of patients at high risk for Candida infections . The mechanisms by which surgery and antibiotics decrease phagocytosis remain to be elucidated.

Ann Clin Lab Sci, 1997 Jul-Aug, 27(4), 282 - 6
Inhibitory effects of seven organosulphur compounds on clinical isolates of Candida species in vitro; Shah DT et al.; Thirty clinical isolates of Candida albicans and 10 other Candida species were tested for susceptibility to 6 substituted dithiocarbamates and one dimercaptosuccinate . Dimethyldithiocarbamate, sodium pyrrolidine dithiocarbamate, and sodium diethyldithiocarbamate showed dose-dependent antifungal activity which was partially reversed by the addition of zinc, copper, or iron sulfate with greatest reversal at 2:1 metal to dithiocarbamate molar ratio . Anaerobiosis also interfered with dithiocarbamate antifungal activity.

Antimicrob Agents Chemother, 1997 Jul, 41(7), 1575 - 8
Effect of granulocyte colony-stimulating factor on the candidacidal activity of polymorphonuclear neutrophils and their collaboration with fluconazole; Natarajan U et al.; The effect of granulocyte colony-stimulating factor (GCSF) treatment of polymorphonuclear neutrophils (PMN) in vitro was studied with respect to their candidacidal activity . The candidacidal activity of PMN was found to be significantly increased when they were pretreated with GCSF . Fluconazole (1 microg/ml) was found to be highly fungistatic (90%) for Candida albicans Sh27 and collaborated with PMN for significantly increased killing . Collaborative killing by PMN significantly increased when they were treated with GCSF before and after fungal exposure . The enhancing activities of GCSF required optimization of the GCSF dose and were thus inoculum and strain dependent.

Antimicrob Agents Chemother, 1997 Jul, 41(7), 1488 - 94
The presence of an R467K amino acid substitution and loss of allelic variation correlate with an azole-resistant lanosterol 14alpha demethylase in Candida albicans; White TC; Azole resistance in the pathogenic yeast Candida albicans is an emerging problem in the human immunodeficiency virus (HIV)-infected population . The target enzyme of the azole drugs is lanosterol 14alpha demethylase (Erg16p), a cytochrome P-450 enzyme in the biosynthetic pathway of ergosterol . Biochemical analysis demonstrates that Erg16p became less susceptible to fluconazole in isolate 13 in a series of isolates from an HIV-infected patient . PCR-single-strand conformation polymorphism (PCR-SSCP) analysis was used to scan for genomic alterations of ERG16 in the isolates that would cause this change in the enzyme in isolate 13 . Alterations near the 3' end of the gene that were identified by PCR-SSCP were confirmed by DNA sequencing . A single amino acid substitution (R467K) that occurred in isolate 13 was identified in both alleles of ERG16 . Allelic differences within the ERG16 gene, in the ERG16 promoter, and in the downstream THR1 gene were eliminated in isolate 13 . The loss of allelic variation in this region of the genome is most likely the result of mitotic recombination or gene conversion . The R467K mutation and loss of allelic variation that occur in isolate 13 are likely responsible for the azole-resistant enzyme activity seen in this and subsequent isolates . The description of R467K represents the first point mutation to be identified within ERG16 of a clinical isolate of C . albicans that alters the fluconazole sensitivity of the enzyme.

Antimicrob Agents Chemother, 1997 Jul, 41(7), 1482 - 7
Increased mRNA levels of ERG16, CDR, and MDR1 correlate with increases in azole resistance in Candida albicans isolates from a patient infected with human immunodeficiency virus; White TC; Resistance to antifungal drugs, specifically azoles such as fluconazole, in the opportunistic yeast Candida albicans has become an increasing problem in human immunodeficiency virus (HIV)-infected individuals . The molecular mechanisms responsible for this resistance have only recently become apparent and can include alterations in the target enzyme of the azole drugs (lanosterol 14alpha demethylase {14DM}), or in various efflux pumps from both the ABC transporter and major facilitator gene families . To determine which of these possible mechanisms was associated with the development of drug resistance in a particular case, mRNA levels have been studied in a series of 17 clinical isolates taken from a single HIV-infected patient over 2 years, during which time the levels of fluconazole resistance of the strain increased over 200-fold . Using Northern blot analysis of steady-state levels of total RNA from these isolates, we observed increased mRNA levels of ERG16 (the 14DM-encoding gene), CDR1 (an ABC transporter), and MDR1 (a major facilitator) in this series . The timing of the increase in mRNA levels of each of these genes correlated with increases in fluconazole resistance of the isolates . Increased mRNA levels were not observed for three other ABC transporters, two other genes in the ergosterol biosynthetic pathway, or the NADPH-cytochrome P-450 oxidoreductase gene that transfers electrons from NADPH to 14DM . Increases in mRNA levels of ERG16 and CDR1 correlated with increased cross-resistance to ketoconazole and itraconazole but not to amphotericin B . A compilation of the genetic alterations identified in this series suggests that resistance develops gradually and is the sum of several different changes, all of which contribute to the final resistant phenotype.

Antimicrob Agents Chemother, 1997 Jul, 41(7), 1455 - 9
Efficacy of D0870 treatment of experimental Candida vaginitis; Fidel PL Jr et al.; In this study, oral administration of the triazole D0870 was compared to oral administration of fluconazole in the treatment of experimental vaginal candidiasis . With an estrogen-dependent murine model of Candida albicans vaginal infection, the effects of D0870 on several isolates, including fluconazole-susceptible and -resistant isolates, were tested . D0870, at doses of 0.5 and 2.5 mg/kg of body weight given once over the course of a 10-day infection, was effective in eradicating vaginitis caused by fluconazole-susceptible laboratory and clinical isolates, respectively . In contrast, a stricter treatment regimen (every 24 to 48 h) with 10 and 25 mg of fluconazole per kg was required to achieve similar reductions in vaginal fungal titers induced by the same isolates . Whereas fluconazole was consistently ineffective in infections induced by fluconazole-resistant isolates, as predicted by in vitro susceptibility tests, D0870 was effective, although a daily regimen of 25 mg/kg was required . Additional studies showed that despite the in vitro activity of D0870 against two clinical Candida glabrata isolates, neither D0870 nor fluconazole was effective at daily doses as high as 100 and 125 mg/kg, respectively . Taken together, although D0870 failed to show efficacy against experimental C . glabrata vaginitis, D0870 was superior to fluconazole in the treatment of experimental C . albicans vaginitis caused by isolates that were either susceptible or resistant to fluconazole.

J Bacteriol, 1997 Jul, 179(13), 4096 - 105
Cloning of the Candida albicans homolog of Saccharomyces cerevisiae GSC1/FKS1 and its involvement in beta-1,3-glucan synthesis; Mio T et al.; Saccharomyces cerevisiae GSC1 (also called FKS1) and GSC2 (also called FKS2) have been identified as the genes for putative catalytic subunits of beta-1,3-glucan synthase . We have cloned three Candida albicans genes, GSC1, GSL1, and GSL2, that have significant sequence homologies with S . cerevisiae GSC1/FKS1, GSC2/FKS2, and the recently identified FKSA of Aspergillus nidulans at both nucleotide and amino acid levels . Like S . cerevisiae Gsc/Fks proteins, none of the predicted products of C . albicans GSC1, GSL1, or GSL2 displayed obvious signal sequences at their N-terminal ends, but each product possessed 10 to 16 potential transmembrane helices with a relatively long cytoplasmic domain in the middle of the protein . Northern blotting demonstrated that C . albicans GSC1 and GSL1 but not GSL2 mRNAs were expressed in the growing yeast-phase cells . Three copies of GSC1 were found in the diploid genome of C . albicans CAI4 . Although we could not establish the null mutation of C . albicans GSC1, disruption of two of the three GSC1 alleles decreased both GSC1 mRNA and cell wall beta-glucan levels by about 50% . The purified C . albicans beta-1,3-glucan synthase was a 210-kDa protein as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and all sequences determined with peptides obtained by lysyl endopeptidase digestion of the 210-kDa protein were found in the deduced amino acid sequence of C . albicans Gsc1p . Furthermore, the monoclonal antibody raised against the purified beta-1,3-glucan synthase specifically reacted with the 210-kDa protein and could immunoprecipitate beta-1,3-glucan synthase activity . These results demonstrate that C . albicans GSC1 is the gene for a subunit of beta-1,3-glucan synthase.

J Infect Dis, 1997 Jul, 176(1), 217 - 26
Induction of protective Th1 responses to Candida albicans by antifungal therapy alone or in combination with an interleukin-4 antagonist; Cenci E et al.; Resistance or susceptibility to disseminated and mucosal Candida albicans infections in mice correlates with the development of protective or nonprotective T helper (Th) cell responses . To determine whether immunomodulatory activity on Th cell functions is an effect beyond that provided by antifungal therapy, mice with disseminated or gastrointestinal infection were treated with amphotericin B or fluconazole and assessed for mortality, fungus burden in the organs, and parameters of Th cell-dependent immunity . Both antimycotics produced protective CD4+ Th1 cell responses, as revealed by increased production of interleukin (IL)-12 and interferon-y, decreased production of IL-4, delayed-type hypersensitivity to fungal antigen, and the disappearance of antigen-specific IgE . Concomitant neutralization of endogenous IL-4 greatly increased the antifungal efficacy and the Th1-promoting activity of both agents . These results indicate that successful antifungal therapy alone or in combination with cytokine antagonists may rely on the induction of an appropriate Th antifungal cell response.

Infect Immun, 1997 Jul, 65(7), 2898 - 903
Effects of pH and salinity on the antimicrobial properties of clavanins; Lee IH et al.; Clavanins are histidine-rich, amidated alpha-helical antimicrobial peptides that were originally isolated from the leukocytes (hemocytes) of a tunicate, Styela clava . The activities of clavanin A amide and clavanin A acid against Escherichia coli, Listeria monocytogenes, and Candida albicans were substantially greater at pH 5.5 than at pH 7.4 . In contrast, clavanin AK, a synthetic variant of clavanin A acid containing 4 histidine-->lysine substitutions exerted substantial activity at both pH 7.4 and pH 5.5 . Each of these three clavanins permeabilized the outer and inner membranes of E . coli very effectively at pH 5.5, but only clavanin AK did so at pH 7.4 . Unlike magainin 1 and cecropin P1, alpha-helical antimicrobial peptides from frog skin and porcine intestine, respectively, clavanins were broadly effective against gram-positive bacteria, including methicillin-resistant Staphylococcus aureus, as well as gram-negative organisms . Because clavanins exert substantial antimicrobial activity in 0.1 to 0.3 M NaCl, they provide templates for designing broad-spectrum peptide antibiotics intended to function in extracellular environments containing normal or elevated NaCl concentrations . The pH-dependent properties of histidine-rich antimicrobial peptides may allow the design of agents that would function selectively in acidic compartments, such as the gastric lumen, or within phagolysosomes.

Infect Immun, 1997 Jul, 65(7), 2663 - 7
Hyperlipoproteinemia enhances susceptibility to acute disseminated Candida albicans infection in low-density-lipoprotein-receptor-deficient mice; Netea MG et al.; Recent studies have suggested the use of lipoproteins as an adjuvant treatment of lethal gram-negative infections . However, other important microorganisms for the etiology of sepsis, such as Candida species, grow better in lipid-rich environments . We investigated the effect of hyperlipoproteinemia on systemic candidiasis in low-density-lipoprotein-receptor-deficient (LDLR-/-) mice, in which the loss of the receptor results in a seven- to ninefold-higher plasma LDL level than that in their wild-type littermates (C57BL/6J) . LDLR-/- mice died earlier, and the outgrowth of Candida albicans in the kidneys and livers of LDLR-/- mice was significantly higher compared with that of controls . After infection, circulating cytokine concentrations were significantly higher in LDLR-/- mice . In vitro, C . albicans grew better in plasma samples of LDLR-/- mice than in control plasma samples and peritoneal macrophages of LDLR-/- mice challenged with heat-killed C . albicans produced more cytokines than did those of controls . This latter phenomenon was probably due to increased binding of yeast cells to macrophages of LDLR-/- mice . These data suggest that hyperlipoproteinemia is deleterious in systemic candidiasis.

J Clin Microbiol, 1997 Jul, 35(7), 1777 - 80
Microsatellite polymorphism in the promoter sequence of the elongation factor 3 gene of Candida albicans as the basis for a typing system; Bretagne S et al.; The polymorphism of a TTC/TTTC microsatellite in the promoter sequence of the elongation factor 3 gene of Candida albicans was investigated by PCR . One primer was fluorescein labeled, and PCR signals were read with an automatic sequencer . Twenty-nine reference strains and 31 independent clinical isolates were studied . Eleven different alleles were identified, giving 16 different profiles among the 60 strains tested, with a discriminatory power of 0.88 . This marker is stable upon subculture, and reproducibility was achieved by automated procedures . When several microsatellite markers are available, many isolates can be rapidly and reproducibly tested for epidemiological questions, such as the prevalence of a given strain in a hospital setting and transmission between patients.

J Clin Microbiol, 1997 Jul, 35(7), 1761 - 5
Variations in fluconazole susceptibility and DNA subtyping of multiple Candida albicans colonies from patients with AIDS and oral candidiasis suffering one or more episodes of infection; Redding SW et al.; Five Candida albicans colonies from each infection in AIDS patients receiving fluconazole therapy for oropharyngeal candidiasis over a 2-year period were evaluated by antifungal susceptibility testing and DNA subtyping, and the results were correlated with clinical response to determine the occurrence of clinically significant selection of more-resistant C . albicans over multiple infections . A total of 534 C . albicans isolates were obtained from 38 patients who exhibited 84 episodes of infection . Antifungal susceptibility testing revealed that the MICs for 93% of the isolates were < or = 8.0 microg/ml and the MICs for 7% of the isolates were > or = 64 microg/ml . DNA subtyping revealed 70 different subtypes, with 78% of patients with one infection exhibiting one DNA subtype and 80% of patients with more than one infection exhibiting multiple DNA subtypes . Also, patients who had multiple infections had lower CD4 counts than those with single infections . Differences between the single-infection group and the multiple-infection group regarding the number of DNA subtypes and CD4 counts were both statistically significant . Of the 74 evaluable infections all were successfully treated with regular-dose (100-mg/day) fluconazole, except for three patients who ultimately responded to higher-dose fluconazole . Only one patient may have shown clinically significant selection of a more-resistant C . albicans strain over multiple courses of treatment . Interestingly, MICs reached only 8.0 microg/ml, even though doses of 400 mg of fluconazole were necessary for clinical cure.

Yeast, 1997 Jun 30, 13(8), 769 - 76
Sequence analysis of the Candida albicans ADE2 gene and physical separation of the two functionally distinct domains of the phosphoribosylaminoimidazole carboxylase; Schmuke JJ et al.; An ADE2 genomic clone from the pathogenic fungus, Candida albicans, was isolated by complementation of an Escherichia coli purK mutant and the gene was analysed by DNA sequencing . A 1707 bp open reading frame was identified encoding a polypeptide of 569 amino acids with significant homology to all the known yeast ADE2 genes . Sequence homology to both the E . coli purE and purK genes suggests that the C . albicans ADE2 gene is the result of an evolutionary fusion . The amino-acid sequence comparison showed that the N-terminal domain of the Ade2 protein has a 52.5% identity to purK, whereas the C-terminal domain has a distinct 64.3% identity to purE . In order to establish the functional relationship of these two regions, deletion mutants of the Ade2 protein were prepared by recombinant expression of the functional domains, which were tested by complementation of their respective E . coli auxotrophs.

Gene, 1997 Jun 19, 192(2), 235 - 40
Sequence and promoter regulation of the PCK1 gene encoding phosphoenolpyruvate carboxykinase of the fungal pathogen Candida albicans; Leuker CE et al.; The PCK1 gene encoding PEP carboxykinase (Pck1) of the fungal pathogen Candida albicans was isolated and sequenced . The deduced Pck1 protein has high homology to ATP-dependent Pck1 proteins in other species, especially to Pck1 of Saccharomyces cerevisiae (70% homology), but not to GTP-dependent Pck1 proteins . PCK1 transcript levels were efficiently repressed by glucose and derepressed (induced) on gluconeogenetic carbon sources . PCK1 regulation occurs on the level of transcription, as demonstrated by a fusion of the PCK1 promoter to the LAC4 reporter gene, yielding derepressed/repressed expression ratios of > 100 . Homologous sequences in the PCK1 promoters of C . albicans and S . cerevisiae were identified . The PCK1 promoter may be useful to efficiently regulate expression and thereby test the function of genes in C . albicans.

FEMS Microbiol Lett, 1997 Jun 15, 151(2), 263 - 8
Molecular analysis of cyp51 from fluconazole-resistant Candida albicans strains; Loffler J et al.; The target enzyme for fluconazole is sterol 14 alpha-demethylase, a cytochrome P450 encoded by cyp51 . One mechanism of fluconazole resistance likely to occur in Candida albicans is through an altered target site . To test this hypothesis DNA sequencing of the cyp51 coding sequence from 19 fluconazole-resistant and 19 fluconazole-sensitive C . albicans was undertaken . A number of point mutations were identified in the resistant isolates which were not present in the sensitive ones: F105L (five), E266D (five), K287R (one), G448G (one), G450E (one), G464S (three) and V488I (one) . These alterations are discussed in the light of a molecular model of the enzyme regarding potential roles in resistance . It was also demonstrated that sequence-specific primers can be employed to identify polymorphisms which may be associated with resistance; diagnostic tests for resistant strains will prove of value in combating this serious clinical problem.

Yeast, 1997 Jun 15, 13(7), 677 - 81
Molecular cloning of a Candida albicans gene (SSB1) coding for a protein related to the Hsp70 family; Maneu V et al.; We have cloned and sequenced a Candida albicans gene (SSB1) encoding a potential member of the heat-shock protein seventy (hsp70) family . The protein encoded by this gene contains 613 amino acids and shows a high degree (85%) of sequence identity to the ssb subfamily (ssb1 and ssb2) of the Saccharomyces cerevisiae hsp70 family . The transcribed mRNA (2.1 kb) is present in similar amounts both in yeast and germ tube cells of C . albicans.

Yonsei Med J, 1997 Jun, 38(3), 178 - 86
Seroreactivities of proteinases of Candida albicans, C . tropicalis, and C . parapsilosis in sera from various Candida species-infected mice; Lee KH et al.; From the culture filtrates of C . albicans, C . tropicalis and C . parapsilosis, proteinases were purified using a series of chromatographic steps consisting of DEAE-Sepharose, Sephacryl S-200 and size-exclusion HPLC which removed contaminating mannoproteins and extraneous proteins . Anti-Candida proteinase antibodies in sera from mice infected with various Candida species were detected using ELISA for serodiagnosis of candidiasis . Three proteinases were blotted by homologous and heterologous anti-proteinase antisera on Western blot analysis . All sera from six Candida species-infected mice were reactive with proteinases of C . albicans, C . tropicalis, and C . parapsilosis, although C . glabrata, C . guilliermondii, and C . krusei did not secrete proteinase . The seroreactivities of proteinase with sera from mice infected with homologous C . albicans and C . tropicalis were higher than those with sera from heterologous Candida species-infected mice . These results suggest that three proteinases have at least one common epitope, but its application for diagnosis of candidiasis should be considered with limits of specificity.

Eur J Epidemiol, 1997 Jun, 13(4), 447 - 50
Prevalence and antifungal susceptibility of vaginal yeasts in outpatients attending a gynecological center in Ancona, Italy; Arzeni D et al.; Between February 1993 and May 1994 we studied the prevalence of fungal vulvovaginitis among women attending the Obstetric and Gynecology Clinic of the University of Ancona . Out of the 222 patients, 18 (8.2%) women had symptomatic vaginitis and 24 (10.8%) were carriers . Candida albicans was the species most frequently isolated (44.2%), followed by Torulopsis glabrata (28%) and Saccharomyces cerevisiae (16.2%), from symptomatic and carrier patients . The activity of acid proteinase was determined for C . albicans isolated from both symptomatic and carrier patients . All 13 carriers showed low activity for aspartyl proteinase (score 1+), while 5 of 6 symptomatic patients showed higher activity (score 2+), with a significant difference (p = 0.026) . In general, isolates of T . glabrata and S . cerevisiae were less susceptible in vitro to fluconazole than isolates of C . albicans . We did not find any differences in fluconazole MIC results among the C . albicans strains isolated from symptomatic and carrier patients . On the other hand, the fluconazole MICs of T . glabrata and S . cerevisiae isolates showed statistically significant differences between symptomatic and carrier patients (p = 0.009 and p = 0.000, respectively) . The differences in proteinase secretion between the isolates from symptomatic and carrier patients suggest a correlation between proteinase production and vaginal candidiasis caused by C . albicans . Torulopsis glabrata, however, was found to be the most common causative agent of vaginitis (7 out 19 episodes), followed by C . albicans (6 out of 19 episodes) . Due to the varying patterns of antifungal susceptibility, mainly to fluconazole for the yeast isolates considered in this study, an in vitro susceptibility testing program might be useful for monitoring the outcome of this infection.

Allerg Immunol (Paris), 1997 Jun, 29(6), 160 - 4
{Diagnosis of delayed type hypersensitivity to Candida albicans . Evaluation of lymphocyte activation by flow cytometry (171 observations)}; Brunet JL et al.; Abnormal delayed-type hypersensitivy to Candida albicans, since it results in an excessive reaction of the immune system, is very difficult to diagnose . This study shows that the syndromic reaction observed after intradermal injection of an extract of Candida albicans, in patients suspected of abnormal delayed-type hypersensitivy to this antigen, is associated with the presence of specific circulating T cells, detectable through cell culture in the presence of Candida albicans . There is a very significant correlation between the clinical symptoms, the cutaneous tests, and the lymphocyte activation tests . This abnormal reactivity essentially involves the CD8 cells.

Diagn Microbiol Infect Dis, 1997 Jun, 28(2), 65 - 7
Evaluation of growth characteristics on blood agar and eosin methylene blue agar for the identification of Candida (Torulopsis) glabrata; Bale MJ et al.; Candida albicans and Candida (Torulopsis) glabrata are the most common species of yeast encountered in the clinical laboratory . In this study, we sought to evaluate simple means of screening cultures for the presence or absence of C . glabrata . Twelve thousand five hundred (12,500) consecutive cultures were evaluated for sufficient yeast growth to warrant identification . When detected (369 isolates), the amount of growth on eosin methylene blue agar (EMB) versus sheep blood agar (BAP) (both incubated in 5% CO2), wet mount morphology, and germ tube production were evaluated . All germ tube-negative yeasts were definitively identified using the Vitek YBC card . Of the 369 yeast isolates included in this study, 225 were C . albicans, 102 C . glabrata, and 42 other Candida species . Growth on EMB was greater than BAP for 92 isolates; all identified as C . glabrata . When EMB growth was equal to or less than BAP, 10 isolates were C . glabrata and 267 were other Candida ssp . An accurate presumptive identification of C . glabrata may be made using the observation of greater growth on EMB versus BAP . When coupled with the germ tube test, the majority of yeast isolates could be identified by these simple methods in our laboratory.

Arzneimittelforschung, 1997 Jun, 47(6), 793 - 6
Augmentation of host defence against bacterial and fungal infections of mice pretreated with the non-pathogenic Escherichia coli strain Nissle 1917; Hockertz S; Escherichia coli strain Nissle 1917 (DSM 6601, Mutaflor) was investigated for its ability to enhance the immune response against bacterial or fungal infections in vivo . Mice were infected intravenously with either 6 x 10(3) colony forming units (cfu) of Listeria monocytogenes bacteria or 5 x 10(5) Candida albicans cells . One day prior to infection, mice were treated orally with four different concentrations of E . coli strain Nissle 1917 (10(6), 10(7), 10(8), and 10(9) viable cells) . Three days after infection with L . monocytogenes or one day after infection with C . albicans, mice were sacrificed and the parasite burden of the main target organs of the respective infection model were examined . The protective effect of E . coli strain Nissle 1917, compared to placebo-treated controls and to mice treated with a dose of 10(4) . Units interferon gamma, is shown as the reduction of viable bacteria in spleen and liver or viable fungi in the kidneys of infected animals, respectively . Orally administered E . coli strain Nissle 1917 reduced Listeria monocyto-genes and Candida albicans in a dose-dependent manner . Treatment with 10(9) cfu of E . coli bacteria led to a reduction of Listeria counts to 7.4% in spleen and 2.4% in liver . A more than 10-fold decrease of viable Candida albicans (residual parasitaemia 6.8%) in the kidneys of the infected animals was also achieved by this E . coli concentration . These results suggest that E . coli strain Nissle 1917 is a potent immunostimulator of bacterial origin with highly protective efficacy against pathogenic bacterial of fungal infections.

FEMS Immunol Med Microbiol, 1997 Jun, 18(2), 105 - 12
Augmented inhibition of growth of Candida albicans by neutrophils in the presence of lactoferrin; Okutomi T et al.; The combined inhibitory effects of neutrophils and lactoferrins on the growth of Candida albicans were examined . Murine or human neutrophils partially inhibited growth of C . albicans when cultured with C . albicans in vitro . The growth inhibition was augmented by a combination of neutrophils and more than 30 microg/ml of bovine lactoferrin or 1 microg/ml of human lactoferrin, concentrations less than 1/10-1/200 their inhibiting concentrations when used alone . The inhibition of C . albicans was also enhanced by combination of neutrophils and bovine apolactoferrin or iron-bound holo-lactoferrin, but not by transferrin . Combination effects of neutrophils and lactoferrin were also observed in a condition where there was no contact between neutrophils and Candida cells . These results suggest that neutrophils inhibit the growth of C . albicans regardless of whether there is direct contact between them and Candida cells: neutrophil growth inhibition effects were augmented in the presence of a physiological concentration of lactoferrin, perhaps through some action of lactoferrin other than chelation of ferric ion.

J Hand Surg {Br}, 1997 Jun, 22(3), 423 - 4
Candida infection of a silicone metacarpophalangeal arthroplasty; Dunkley AB et al.; Fungal infections following joint arthroplasty are extremely rare . Only 16 cases of Candida prosthetic infections have been reported, involving the hip, knee or shoulder joints . We report a case of a silicone metacarpophalangeal joint replacement complicated by a Candida albicans infection.

Biol Pharm Bull, 1997 Jun, 20(6), 637 - 40
Anti Candida activity of induced transferrin in mice immunized with inactivated Candida albicans; Watanabe T et al.; Mice immunized with formalin-killed Candida albicans were resistant to challenge by a lethal amount of viable C . albicans . The growth-inhibitory activity to C . albicans was detected in sera from the immunized mice, and was inhibited by the addition of anti-transferrin antibody or ferric sulfate . Both the amount of transferrin and the unsaturated iron-binding capacity (UIBC) in the serum were significantly increased, indicating that apo-transferrin increased in the immunized mice . Moreover, the intraperitoneal administration of apo-transferrin enhanced the protection from the Candida infection in vivo.

J Endocrinol, 1997 Jun, 153(3), 475 - 83
Modulation of human neutrophil function in vitro by gastrin; De la Fuente M et al.; We have studied the effects in vitro of gastrin-17 and gastrin-34, at concentrations from 10(-14) M to 10(-6) M, on several of the functions of peripheral blood human neutrophils, i.e . adherence to substrate, mobility (spontaneous and directed by a chemical gradient or chemotaxis), ingestion of inert particles (latex beads) and cells (Candida albicans) and superoxide anion production . Both gastrins inhibited several steps of the phagocytic process of human neutrophils, such as mobility and ingestion . By contrast, these peptides increased adherence and had no effect on superoxide anion production . In general, these effects were significant at peptide concentrations between 10(-12) M and 10(-8) M with a maximal effect at 10(-10) M . In addition, gastrin peptides induced a significant increase in intracellular cAMP levels at 30, 60 and 120 s . Moreover, the inhibitory effect of gastrin-17 on the ingestion capacity of neutrophils (latex bead phagocytosis) was similar to that obtained with EGTA, a well-known extracellular calcium chelating compound . Gastrin-17 was found to inhibit completely the stimulation of latex bead phagocytosis in neutrophils caused by the calcium ionophore A23187 . These results suggest that gastrin is a negative modulator of the phagocytic process of human neutrophils, and that this effect might involve an increase in intracellular cAMP levels and a decrease in calcium entry into the cells.

Pediatr Nephrol, 1997 Jun, 11(3), 325 - 7
Peritonitis in continuous ambulatory peritoneal dialysis in children living in Saudi Arabia; Mirza K et al.; The clinical aspects of peritonitis and catheter infections were reviewed in 64 children on continuous ambulatory peritoneal dialysis living in Saudi Arabia over a period of 6 years . Peritonitis occurred in 41 children (64%) . The mean time from starting dialysis to the first episode of peritonitis was 7.2 months . The incidence of peritonitis was 1 episode in 9 treatment months . Gram-negative organisms were responsible for the majority of episodes (42%), followed by Gram-positive organisms (20%), and Candida albicans (6%); 32% were culture negative . Recurrent peritonitis was present in 20 cases . Catheter was replaced in 24 patients: 44% due to recurrent peritonitis . Peritoneal membrane loss occurred in 7 patients, 3 had Candida peritonitis and 3 had recurrent peritonitis due to Pseudomonas . The mortality rate was 4.6% but none of the deaths were related to peritonitis or dialysis.

Scand J Immunol, 1997 Jun, 45(6), 596 - 604
A human in vitro granuloma model using heat killed Candida albicans cells immobilized on plastic culture wells; Heinemann DE et al.; A new model for studying the initial events of granuloma formation in vitro is presented using heat killed Candida albicans immobilized on the surface of plastic culture wells . Human monocytes were induced to accumulate and to proliferate, forming multinucleated giant cells (MGC) and epitheloid cells within 4 days of culture . Tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta and IL-6 were detected in culture supernatants . These monokines, and additionally macrophage colony stimulating factor (M-CSF), were also detected immunocytochemically . The granuloma formation was inhibited by Dexamethasone (Dex), Pentoxifylline (POF), or interferon-gamma (IFN-gamma) in a dose-dependent manner . Antibodies to M-CSF reduced the granuloma formation to a great extent with a striking reduction of monocyte proliferation . Using antibodies to TNF-alpha the authors found a complete inhibition of the granuloma including MGC formation and monocyte proliferation.

Clin Infect Dis, 1997 Jun, 24(6), 1172 - 7
Infectious ocular complications in orthotopic liver transplant patients; Papanicolaou GA et al.; We report the frequency and type of infectious ocular complications following orthotopic liver transplantation (OLT) and review diagnostic and therapeutic strategies . During the period September 1988 through November 1994, 684 patients underwent OLT at Mount Sinai Hospital (New York) . Nine orthotopic liver transplant patients (1.3%) developed ocular infections: Candida albicans endophthalmitis (2), Aspergillus fumigatus endophthalmitis (1), cytomegalovirus retinitis (4), herpes simplex virus keratitis (1), and varicella-zoster virus panophthalmitis (1) . The mean time from OLT to ocular symptoms was 42 days for patients with fungal infections and 128 days for patients with viral infections . Blurred vision was the commonest symptom (five of nine cases) . The mean duration of follow-up was 2 years (range, 33 days to 5 years) . Permanent loss of vision occurred in three patients, five had improvement in visual acuity, and one died of disseminated aspergillosis 33 days after OLT . Infectious ocular complications following OLT may occur as isolated events or with disseminated disease . Fungal infections occur earlier (mean, 42 days after OLT) than viral infections (mean, 4 months after OLT) . The clinical presentation may be atypical; aggressive vitreoretinal procedures and serial examinations may be required to establish the diagnosis . Cytomegalovirus retinitis in orthotopic liver transplant patients may not require life-long maintenance therapy with antiviral agents.

J Bacteriol, 1997 Jun, 179(12), 3837 - 44
The WH11 gene of Candida albicans is regulated in two distinct developmental programs through the same transcription activation sequences; Srikantha T et al.; Candida albicans strain WO-1 undergoes two developmental programs, the bud-hypha transition and high-frequency phenotypic switching in the form of the white-opaque transition . The WH11 gene is expressed in the white budding phase but is inactive in the white hyphal phase and in the opaque budding phase . WH11 expression, therefore, is regulated in the two developmental programs . Through fusions between deletion derivatives of the WH11 promoter and the newly developed Renilla reniformis luciferase, the WH11 promoter has been characterized in the two developmental programs . Three transcription activation sequences, two strong and one weak, are necessary for the full expression of WH11 in the white budding phase, but no negative regulatory sequences were revealed as playing a role in either the white hyphal phase or the opaque budding phase . These results suggest that regulation is solely through activation in the white budding phase and the same mechanism, therefore, is involved in regulating the differential expression of WH11 in the alternative white and opaque phases of switching and the budding and hyphal phases of dimorphism.

Microbiol Mol Biol Rev, 1997 Jun, 61(2), 170 - 92
Macrophages in resistance to candidiasis; Vazquez-Torres A et al.; Candida albicans, an increasingly common opportunistic pathogenic fungus, frequently causes disease in immunodeficient but not immunocompetent hosts . Clarifying the role of the phagocytic cells that participate in resistance to candidiasis not only is basic to understanding how the host copes with this dimorphic pathogen but also will expedite the development of innovative prophylactic and therapeutic approaches for treating the multiple clinical presentations that candidiasis encompasses . In this review, we present evidence that a diverse population of mononuclear phagocytes, in different states of activation and differentiation and from a variety of host species, can phagocytize C . albicans blastoconidia via an array of opsonic and nonopsonic mechanisms and can kill C . albicans blastoconidia and hyphae by means of oxygen-dependent and -independent mechanisms . Reactive nitrogen intermediates should now be added to the well-established candidacidal reactive oxygen intermediates of macrophages . Furthermore, what were thought to be two independent pathways, i.e., nitric oxide and superoxide anion, have now been shown to combine to form a potent macrophage candidacidal molecule, peroxynitrite . In contrast to monocytes and neutrophils, which are important in resistance to early stages of C . albicans infections, more differentiated macrophages activated by cytokines such as gamma interferon participate in the acquired resistance of hosts with C . albicans-specific, cell-mediated immunity . Evidence presented in this review demonstrates that mononuclear phagocytes, in some instances in the absence of other professional phagocytes such as neutrophils, play an import role in resistance to systemic and mucosal candidiasis.

J Infect Dis, 1997 Jun, 175(6), 1467 - 76
Iron overload alters innate and T helper cell responses to Candida albicans in mice; Mencacci A et al.; The effect of iron overload on susceptibility of mice to Candida albicans infection and on the type of T helper (Th) immunity elicited was investigated . Iron overload greatly increased susceptibility to disseminated infection with low-virulence C . albicans cells of exogenous origin . The candidacidal activity and the ability to release nitric oxide and bioactive interleukin (IL)-12 were greatly impaired in neutrophils and macrophages from infected mice . CD4 T cells from spleens of iron-overloaded mice were found to produce high levels of IL-4 and IL-10 and low levels of interferon-gamma . Treatment of iron-overloaded mice with the iron chelator, deferoxamine, resulted in the cure of mice from infection, restored the antifungal effector and immunomodulatory functions of the phagocytic cells, and allowed the occurrence of CD4 Th1 protective antifungal responses . These data indicate that iron overload may negatively affect CD4 Th1 development in mice with candidiasis, a function efficiently restored by therapy with deferoxamine.

Antimicrob Agents Chemother, 1997 Jun, 41(6), 1392 - 5
Antifungal pharmacodynamic characteristics of fluconazole and amphotericin B tested against Candida albicans; Klepser ME et al.; Time-kill curves were determined for three isolates of Candida albicans tested against fluconazole and amphotericin B at multiples of the MIC . Fluconazole produced fungistatic activity, with concentration-related growth effects observed over a narrow range of concentrations . Amphotericin B exhibited fungicidal activity, with enhancement of activity over a broader range of concentrations.

Antimicrob Agents Chemother, 1997 Jun, 41(6), 1345 - 8
Combination therapy with amphotericin B and fluconazole against invasive candidiasis in neutropenic-mouse and infective-endocarditis rabbit models; Sanati H et al.; Although there are an increasing number of new antifungal agents available, the morbidity and mortality due to invasive mycoses remain high . The high rates of polyene toxicities and the development of azole resistance have raised the issue of using antifungal agents of these classes in combination, despite theoretical concerns regarding antagonism between such agents . This study was designed to evaluate the in vivo efficacy of combined therapy with amphotericin B and fluconazole against Candida albicans . Two distinct animal models were used in this study: a neutropenic-mouse model of hematogenously disseminated candidiasis and the infective-endocarditis rabbit model . Treatment efficacy was assessed by determining reductions in mortality as well as decreases in tissue fungal densities . In the neutropenic-mouse model, amphotericin B, as well as combination therapy, significantly prolonged survival compared to untreated controls (P < 10(-5) and P = 0.001, respectively) . The fungal densities in the kidneys of neutropenic mice were significantly reduced with either amphotericin B monotherapy or amphotericin B-fluconazole combined therapy compared to those of controls (P < 10(-6)) . Fluconazole monotherapy also reduced fungal densities in the kidneys; however, this decrease was not statistically significant (P = 0.17) . In contrast, treatment with either fluconazole alone or combined with amphotericin B (but not amphotericin B monotherapy) significantly decreased fungal densities in the brain (P = 0.025) . In the rabbit endocarditis model, amphotericin B monotherapy or combined therapy significantly decreased fungal densities in cardiac vegetations (P < 0.01 versus the controls) . Although no significant antagonism was seen when fluconazole was given in combination with amphotericin B, combination therapy did not augment the antifungal activity of amphotericin B.

J Clin Microbiol, 1997 Jun, 35(6), 1473 - 6
Effects of incubation time and buffer concentration on in vitro activities of antifungal agents against Candida albicans; Tornatore MA et al.; Nine selected isolates of Candida albicans were tested for their susceptibilities to amphotericin B and fluconazole by using three methods to assess the effect of incubation time and buffer concentration . By using a microdilution method with 0.0165 M 3-(N-morpholino)propanesulfonic acid (MOPS) and a 24-h incubation time, all of the isolates were found to be susceptible to amphotericin B and fluconazole . After 48 h of incubation, all isolates were still susceptible to amphotericin B . Seven of the nine isolates were resistant to fluconazole, and for the remaining two isolates, MICs increased by fourfold or more but the isolates remained susceptible (MIC, < or = 10 microg/ml) . The nine isolates, along with three control strains, were further tested against amphotericin B and fluconazole by a standard broth macrodilution method with both 0.165 and 0.0165 M MOPS . The susceptibility results for fluconazole by the broth macrodilution method with the lower MOPS concentration correlated with the results of the 24-h broth microdilution method for determination of susceptibility or resistance in eight of nine tests and with the results of the 48 h broth microdilution method in three of nine tests . The results of the broth macrodilution method with the standard MOPS concentration did not correlate with any of the results obtained by the 24-h broth microdilution but correlated with results of seven of nine tests by the 48-h broth microdilution method . All nine test strains appeared to be susceptible when they were examined by a flow cytometric method . For clinical yeast susceptibility testing in microdilution panels, the 0.0165 M MOPS concentration combined with 24 h of incubation appeared to be the method of choice . The lower MOPS concentration may also be a useful modification to the tentative broth macrodilution method of the National Committee for Clinical Laboratory Standards . Use of the higher buffer concentration or longer incubation time may lead to false in vitro resistance for agents like fluconazole.

J Clin Microbiol, 1997 Jun, 35(6), 1454 - 9
Rapid identification of Candida species in blood cultures by a clinically useful PCR method; Shin JH et al.; Widespread use of fluconazole for the prophylaxis and treatment of candidiasis has led to a reduction in the number of cases of candidemia caused by Candida albicans but has also resulted in the emergence of candidemias caused by innately fluconazole-resistant, non-C . albicans Candida species . Given the fulminant and rapidly fatal outcome of acute disseminated candidiasis, rapid identification of newly emerging Candida species in blood culture is critical for the implementation of appropriately targeted antifungal drug therapy . Therefore, we used a PCR-based assay to rapidly identify Candida species from positive blood culture bottles . This assay used fungus-specific, universal primers for DNA amplification and species-specific probes to identify C . albicans, C . krusei, C . parapsilosis, C . tropicalis, or C . glabrata amplicons . It also used a simpler and more rapid (1.5-h) sample preparation technique than those described previously and used detergent, heat, and mechanical breakage to recover Candida species DNA from blood cultures . A simple and rapid (3.5-h) enzyme immunosorbent assay (EIA)-based format was then used for amplicon detection . One hundred fifty blood culture bottles, including 73 positive blood culture bottle sets (aerobic and anaerobic) from 31 patients with candidemia, were tested . The combined PCR and EIA methods (PCR-EIA) correctly identified all Candida species in 73 blood culture bottle sets, including bottles containing bacteria coisolated with yeasts and 3 cultures of samples from patients with mixed candidemias originally identified as single-species infections by routine phenotypic identification methods . Species identification time was reduced from a mean of 3.5 days by routine phenotypic methods to 7 h by the PCR-EIA method . No false-positive results were obtained for patients with bacteremias (n = 18), artificially produced non-Candida fungemias (n = 3), or bottles with no growth (n = 20) . Analytical sensitivity was 1 cell per 2-microl sample . This method is simpler and more rapid than previously described molecular identification methods, can identify all five of the most medically important Candida species, and has the potential to be automated for use in the clinical microbiology laboratory.

J Clin Microbiol, 1997 Jun, 35(6), 1332 - 6
Comparative analysis of genetic variability among Candida albicans isolates from different geographic locales by three genotypic methods; Clemons KV et al.; The objective of the present study was to conduct a comparative genotypic analysis of Candida albicans isolates from the United States, Europe, and Southeast Asia to determine whether differences between isolates might be associated with geographic locations . The genotypes of 86 unrelated isolates of C . albicans (from the United States and Europe) and 26 isolates from Singapore were examined by three DNA typing methods . Computer-assisted methods were used to analyze the gel patterns for all isolates . A dendrogram based on the overall similarity of the patterns obtained by restriction endonuclease analysis (REA) with EcoRI clustered the U.S . and European isolates into two major groups (groups A and B) . The Singaporean isolates demonstrated unique REA profiles, with nine isolates having both or neither of the REA-characteristic 3.7- and 4.2-kb bands present in groups A and B . By REA profiles, the Singaporean isolates were related to each other with similarity values (S(AB)s) of > 0.80, but only one isolate mixed with the U.S . and European isolates at this S(AB) (an arbitrary threshold for genetic similarity) . Randomly amplified polymorphic DNA (RAPD) analysis generated DNA profiles that clustered the C . albicans isolates into approximately the same number of distinct typing groups as REA . However, isolates identical to each other by REA were generally different from each other by RAPD analysis . In a composite dendrogram prepared from the results obtained by RAPD analysis, the isolates from the United States and Europe clustered in major groups with S(AB)s of > 0.85, while Singaporean isolates connected to these clusters at S(AB)s of > or = 0.75 . Pulsed-field gel electrophoresis was less discriminatory, discerning about one-third as many distinct subtypes as REA or RAPD analysis; the Singaporean isolates were distributed randomly with the U.S . and European isolates . These results suggest that a high degree of genetic diversity exists between C . albicans isolates from Southeast Asia and those from the United States and Europe.

Biochem J, 1997 May 15, 324 ( Pt 1), 329 - 39
Molecular cloning and expression of the Candida albicans TOP2 gene allows study of fungal DNA topoisomerase II inhibitors in yeast; Keller BA et al.; Candida albicans topoisomerase II, encoded by the TOP2 gene, mediates chromosome segregation by a double-strand DNA break mechanism and is a potential target for anti-fungal therapy . In this paper, we report the characterization of the C . albicans TOP2 gene and its use to develop a yeast system that allows the identification and study of anti-fungal topoisomerase II inhibitors in vivo . The gene, specifying a 1461-residue polypeptide with only 40% identity with human topoisomerase IIalpha and beta isoforms, was isolated from C . albicans on a 6.3 kb EcoRI fragment that mapped to chromosome 4 . It was used to construct a plasmid in which TOP2 expresses a recombinant enzyme (residues 57-1461 of C . albicans topoisomerase II fused to the first five residues of Saccharomyces cerevisiae topoisomerase II) under the control of a galactose-inducible promoter . The plasmid rescued the lethal phenotype of a temperature-sensitive S . cerevisiae DNA topoisomerase II mutant allowing growth at 35 degrees C . Yeast cells, bearing ISE2 permeability and rad52 double-strand-break-repair mutations the growth of which at 35 degrees C was dependent on C . albicans topoisomerase II, were killed by the known topoisomerase II inhibitors amsacrine and doxorubicin . Parallel experiments in yeast expressing human topoisomerase IIalpha allowed the relative sensitivities of the fungal and host topoisomerases to be examined in the same genetic background . To compare the killing in vivo with drug inhibition in vitro, the recombinant C . albicans topoisomerase II protein was expressed and purified to near-homogeneity from S . cerevisiae yielding a 160 kDa polypeptide that displayed the expected ATP-dependent DNA-relaxation and DNA-decatenation activities . The enzyme, whether examined in vitro or complementing in S . cerevisiae, was comparably sensitive to amsacrine and doxorubicin . Our results suggest that potential topoisomerase II-targeting anti-fungal inhibitors can be identified and studied in S . cerevisiae.

FEBS Lett, 1997 May 12, 408(1), 89 - 93
Biogenesis of Candida albicans Can1 permease expressed in Saccharomyces cerevisiae; Matijekova A et al.; The Candida albicans CAN1 gene, encoding a high-affinity permease for arginine, lysine and histidine, was tagged at its C-terminus with a c-myc epitope and introduced into strains of Saccharomyces cerevisiae lacking basic amino acid permeases . The expression levels of Ca-Can1p were influenced by the available nitrogen source, being almost negligible when cells were grown in the presence of ammonia . Ca-Can1p was shown to follow the secretory pathway in S . cerevisiae . Ca-Can1p activity was not detected in a secretion-defective sec1-1 mutant grown at a non-permissive temperature . Shr3p, an ER protein that participates in the biogenesis of amino acid permeases was also required for the functional expression of Ca-Can1p . The shr3 mutation does not affect the affinity for substrate but does decrease the number of Can1p molecules reaching the plasma membrane.

J Med Chem, 1997 May 9, 40(10), 1422 - 38
Conformationally constrained {p-(omega-aminoalkyl)phenacetyl}-L-seryl-L-lysyl dipeptide amides as potent peptidomimetic inhibitors of Candida albicans and human myristoyl-CoA:protein N-myristoyl transferase; Nagarajan SR et al.; MyristoylCoA:protein N-myristoyltransferase (NMT) covalently attaches the 14-carbon saturated fatty acid myristate, via an amide bond, to the N-terminal glycine residues of a variety of cellular proteins . Genetic studies have shown that NMT is essential for the viability of the principal fungal pathogens which cause systemic infection in immunosuppressed humans and hence is a target for development of fungicidal drugs . We have generated a class of potent peptidomimetic inhibitors of the NMT from one such fungal pathogen, Candida albicans . The N-terminal tetrapeptide from a substrate analog inhibitor, ALYASKL-NH2, was replaced with an omega-aminoalkanoyl moiety having an optimal 11-carbon chain for inhibition (11-aminoundecanoyl-SKL-NH2, 3a, IC50 = 1.2 +/- 0.14 microM) . A series of replacements for the C-terminal Leu established that residues containing a lipophilic side chain were most effective, with cyclohexylalanine having the greatest potency (3g, IC50 = 0.36 +/- 0.06 microM) . Removal of the carboxamide moiety led to a metabolically stable dipeptide inhibitor containing an N-(cyclohexylethyl)lysinamide (17e, IC50 = 0.11 +/- 0.03 microM) . Partial rigidification of the flexible aminoundecanoyl chain produced the dipeptide p-(omega-aminohexyl)phenacetyl-L-seryl-L-lysyl-N-(cyclohexyleth yl)amide (26b, IC50 = 0.11 +/- 0.04 microM) . Subsequent incorporation of an alpha-methyl substituent into 26b provided the dipeptide analog {2-{p-(omega-aminohexyl)phenyl}propionyl}-L-seryl-L-lysyl-N-(cyclohex ylethyl)amide, a very potent inhibitor (48, IC50 = 0.043 +/- 0.006 microM), which retained the three essential elements required for recognition by the acyl transferase's peptide binding site.

J Biol Chem, 1997 May 2, 272(18), 11874 - 80
Scanning alanine mutagenesis and de-peptidization of a Candida albicans myristoyl-CoA:protein N-myristoyltransferase octapeptide substrate reveals three elements critical for molecular recognition; McWherter CA et al.; Candida albicans produces a single myristoyl-CoA:protein N-myristoyltransferase (Nmt) that is essential for its viability . An ADP-ribosylation factor (Arf) is included among the few cellular protein substrates of this enzyme . An octapeptide (GLYASKLS-NH2) derived from a N-terminal Arf sequence was used as the starting point to identify elements critical for recognition by the acyltransferases's peptide-binding site . In vitro kinetic studies, employing purified Nmt and a panel of peptides with single Ala substitutions at each position of GLYASKLS-NH2, established that its Gly1, Ser5, and Lys6 residues play predominant roles in binding . ALYASKLS-NH2 was found to be an inhibitor competitive for peptide (Ki = 15.3 +/- 6.4 microM) and noncompetitive for myristoyl-CoA (Ki = 31.2 +/- 0.7 microM) . A survey of 26 derivatives of this inhibitor, representing (i) a complete alanine scan, (ii) progressive C-terminal truncations, and (iii) manipulation of the physical-chemical properties of its residues 1, 5, and 6, confirmed the important stereochemical requirements for the N-terminal amine, the beta-hydroxyl of Ser5, and the epsilon-amino group of Lys6 . Remarkably, replacement of the the N-terminal tetrapeptide of ALYASKLS-NH2 with an 11-aminoundecanoyl group produced a competitive inhibitor, 11-aminoundecanoyl-SKLS-NH2, that was 38-fold more potent (Ki = 0.40 +/- 0.03 microM) than the starting octapeptide . Removing the primary amine (undecanoyl-SKLS-NH2), or replacing it with a methyl group (dodecanoyl-SKLS-NH2), resulted in 26- and 34-fold increases in IC50, confirming the important contribution of the amine to recognition . Removal of LeuSer from the C terminus (11-aminoundecanoyl-SK-NH2) yielded a competitive dipeptide inhibitor with a Ki (11.7 +/- 0.4 microM) equivalent to that of the starting octapeptide, ALYASKLS-NH2 . Substitution of Ser with homoserine, cis-4-hydroxyproline, or tyrosine reduces potency by 3-70-fold, emphasizing the requirement for proper presentation of the hydroxyl group in the dipeptide inhibitor . Substituting D- for L-Lys decreases its inhibitory activity >100-fold, while deletion of the epsilon-amino group (Nle) or masking its charge (epsilon-N-acetyl-lysine) produces 4-7-fold attenuations . L-His, but not its D-isomer, can fully substitute for L-Lys, producing a competitive dipeptide inhibitor with similar potency (Ki = 11.9 +/- 1.0 microM) . 11-Aminoundecanoyl-SK-NH2 and 11-aminoundecanoyl-SH-NH2 establish that a simple alkyl backbone can maintain an appropriate distance between three elements critical for recognition by the fungal enzyme's peptide-binding site: a simple omega-terminal amino group, a beta-hydroxyl, and an epsilon-amino group or an imidazole . These compounds contain one peptide bond and two chiral centers, suggesting that it may be feasible to incorporate these elements of recognition, or functionally equivalent mimics, into a fully de-peptidized Nmt inhibitor.

J Med Vet Mycol, 1997 May-Jun, 35(3), 219 - 24
Membrane changes associated with the early stages of apoptosis in HEp-2 cells decrease susceptibility to adherence by Candida albicans; Cotter G et al.; The aim of the present work was to establish whether apoptotic HEp-2 cells demonstrated an altered susceptibility to adherence by the pathogenic yeast Candida albicans . Cultures of HEp-2 cells were treated with concentrations of three chemotherapeutic agents sufficient to induce cell death by apoptosis and this was confirmed by microscopic examination and by the loss of membrane asymmetry (as indicated by phosphatidylserine externalization) and the fragmentation of nuclear DNA into distinct subunits . Cells in the early stages of apoptosis demonstrated a reduced susceptibility to adherence by a number of strains of C . albicans . A correlation between the decrease in yeast adherence and the loss of membrane asymmetry, associated with the early stages of apoptosis, was established.

J Med Vet Mycol, 1997 May-Jun, 35(3), 197 - 203
Increased tissue resistance in the nude mouse against Candida albicans without altering strain-dependent differences in susceptibility; Fulurija A et al.; Strain differences in tissue responses to infection with Candida albicans were examined in nude mice having susceptible (CBA/CaH) and resistant (BALB/c) parentage . Homozygous (nu/nu) mice of both strains were more resistant to systemic infection with C . albicans than heterozygous (nul+) littermates as indicated by a reduction in both the severity of tissue damage and colony counts in the brain and kidney . However, the tissue lesions in nu/nu CBA/CaH mice were markedly more severe than those in nu/nu mice with the BALB/c background . This pattern was reflected in the greater fungal burden in the CBA/CaH strain . Analysis of cDNA from infected tissues using a competitive polymerase chain reaction excluded interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha), and interleukin 6 (IL-6) as mediators of the enhanced resistance of the nude mice . The results confirm that the different patterns of lesion severity in BALB/c and CBA/CaH mice do not involve T lymphocyte-mediated pathology, and are consistent with the hypothesis that strain-dependent tissue damage is not dependent on the effector function of macrophages or their precursors.

J Med Vet Mycol, 1997 May-Jun, 35(3), 173 - 9
Molecular cloning of a Rho family, CDC42Ca gene from Candida albicans and its mRNA expression changes during morphogenesis; Mirbod F et al.; The small GTP-binding protein family regulates various cell functions in mammalian and yeast cells . In the yeast Saccharomyces cerevisiae it has been known to be involved in vegetative growth . As an initial attempt to explore the involvement of CDC42, a member of this family, in the regulation of morphological changes in Candida albicans, we isolated a gene encoding this protein (CDC42Ca) from this fungus . The sequence of isolated gene revealed an open reading frame of 570 nucleotides with the potential to encode a protein of 190 amino acids with a predicted molecular weight of 20.5 kDa . The deduced amino acid sequence was highly homologous to CDC42s from yeast (87.8%), human (76.4%) and Caenorhabditis elegans (73.7%) . The CDC42Ca mRNA level showed a transient increase with a peak at 2 h after the fresh medium shift (28 degrees C) when cells synchronously formed buds, whereas it displayed a gradual increase up to 12 h after the medium shift (37 degrees C) with elongation of germ tubes . This suggests that CDC42 may play a role in the bud emergence and also germ tube formation in C . albicans.

Eur J Clin Microbiol Infect Dis, 1997 May, 16(5), 346 - 50
Semiquantitative polymerase chain reaction enzyme immunoassay for diagnosis of disseminated candidiasis; Burnie JP et al.; A polymerase chain reaction enzyme immunoassay (PCR-EIA) was developed for the semiquantitative of circulating candidal DNA in disseminated candidiasis due to Candida albicans . Polymerase chain reaction was based on primers from the internal transcribed ribosomal region . Binding of the product to a streptavidin-coated microtitration plate was mediated by a biotinylated capture probe . The product was digoxigenylated during PCR; this was the tag to which antibody was bound in the subsequent EIA . The optical density (OD) endpoint was < 0.1 in 15 sera from patients with no evidence of candidal infection (group 1) and in 13 of the 16 sera from colonized patients (group 2); it was > 0.1 in the other three sera from group 2 blood culture-negative patients who required intravenous amphotericin B for cure . The OD was positive in 28 patients with disseminated candidiasis (group 3), defined as positive blood cultures and successful treatment with amphotericin B (n = 11), positive blood culture confirmed at autopsy (n = 11), or negative blood culture first proven at necropsy (n = 6) . In patients from whom multiple samples were available, recovery correlated with an optical density of < 0.1 by day 4 in four patients and by day 13 in the rest . In the five patients with fatal outcome from whom multiple samples were available, the mean OD rose from 0.174 to 0.668 . Samples seeded with Candida albicans blastoconidia demonstrated that on OD of 0.220 was equivalent to 10 cfu . Assay of the group 3 sera by a commercial antigen detection test gave a corresponding sensitivity of 60% which rose to 67.9% when an in-house reverse passive latex agglutination test was used.

J Surg Res, 1997 May, 69(2), 399 - 407
Candida infection following severe trauma exacerbates Th2 cytokines and increases mortality; Mack VE et al.; Following trauma, there is an increase of Th2 cytokines (IL-4, IL-6, and IL-10) and a decrease in Th1 cytokines (IFN-gamma and IL-2) that may account for impaired cellular immunity . However, the functional significance of a dominant Th2 pattern to the host remains unclear . The aim of this study was to evaluate whether Candida albicans (CA) sepsis in the setting of a Th2 response to trauma leads to increased mortality and to examine the mediators involved . Female BALB/c mice were randomized (12 per group) to receive no injury (C); trauma, consisting of a combined femur fracture and 40% total blood loss (T); no injury plus CA infection (C+CA); and CA infection 1 week following trauma (T+CA) . Survival was then followed for 3 weeks . In a separate study, mice were treated as above (5 per group) and sacrificed . Harvested splenocytes were evaluated for concanavalin A-stimulated cytokine production and liver and kidney homogenates were plated to evaluate CA growth per organ and examined histologically . Candida infection at 1 week following trauma resulted in significantly increased mortality compared to infected controls . Furthermore, the Th2 dominant cytokine pattern was significantly augmented in the presence of CA infection in both C+CA and T+CA groups . Additional analysis showed significant growth of CA in liver and kidney homogenates from T+CA compared to C+CA mice . These results suggest that injured and infected mice demonstrate augmentation of Th2 dominant responses above that of injury or infection alone, as well as a decreased ability to clear Candida which may partially explain the increase in mortality observed . Therapies designed to neutralize Th2 cytokines or augment Th1 cytokines may prove beneficial in the setting of sepsis following trauma.

Arzneimittelforschung, 1997 May, 47(5), 667 - 70
Antifungal activity of organic and organometallic derivatives of benzimidazole and benzothiazole; Kucukbay H et al.; Forty organic or organometallic derivatives of benzimidazole and benzothiazole and five rhodium(I) and ruthenium(II) complexes were evaluated for their in vitro antifungal activity against Candida albicans . Four of the tested compounds, the rhodium containing compounds 30, 31, 32 and 33, were found effective at the minimum inhibitory concentrations(MICs) between 400-600 micrograms/ml.

Clin Neuropathol, 1997 May-Jun, 16(3), 143 - 6
Primary Candida albicans empyema associated with epidural hematomas in craniocervical junction; Duffner F et al.; The case of a 27-year-old patient with chronic candida empyema in the craniocervical junction is presented . Occlusive hydrocephalus at admittance, primary subdural candida empyema, and recurrent epidural bleedings are the outstanding features in the clinical course . Despite intact immunity this patient acquired primary candidosis of CNS . Pathological changes in dura, ventricular system, and CSF required multiple shunt revisions . Antimycotic therapy was performed with a combination of 3 antimycotics . The clinical improvement was prolonged by several complications.

Gen Pharmacol, 1997 May, 28(5), 797 - 804
A new substance (Yoshixol) with an interesting antibiotic mechanism from wood oil of Japanese traditional tree (Kiso-Hinoki), Chamaecyparis obtusa; Koyama S et al.; 1 . A neutral wood oil was extracted from Chamaecyparis obtusa (Kiso-Hinoki), which has been trusted nationally and preserved historically in the central part of Japan (Kiso, Nagano) . 2 . Hinokitiol, or thujaplicin (C10H12O2), which has been believed to exist in Cupressaceae, was not found in this neutral wood oil . Some differences between the extracting processes of the natural products are discussed . 3 . A new chemical substance (Yoshixol, 4,4-dimethyl-6-methylene-2-cyclohexen-1-one) was simulated by several criteria (details in the text) as a major candidate of the neutral wood oil from Chamaecyparis obtusa . Thus, Yoshixol was newly synthesized . 4 . The antibiotic effects of hinokitiol, the neutral wood oil and Yoshixol on methicillin-resistant Staphylococcus aureus (MRSA) were examined bacteriologically and morphologically . 5 . All of the aforementioned three test materials showed complete antibiotic effects on MRSA by the bacteriological examination . However, the morphological findings showed entirely different aspects of cell death . 6 . Hinokitiol caused an aggregative, degenerative and/or necrotic aspect, but the neutral wood oil and Yoshixol produced characteristic aspects: separation of contacted cells, blebbing, bugging-like eruption, formation of granules and an extensive reduction of individual cell size of MRSA . 7 . Yoshixol was able to enhance those antibiotic effects on MRSA distinctly more than the neutral wood oil . 8 . Yoshixol also showed a strong antibiotic effect on Escherichia coli, Mycobacterium chelonei, Pseudomonas aureginosa and Candida albicans . Morphological observations of those bacilli after Yoshixol revealed characteristic aspects of separation of contacted cells, bugging-like swelling, granulation, ballooning and reduction of cell size . 9 . A possible mechanism of Yoshixol is discussed in regard to a molecular orbital theory on the basis of its electron orbits and to a thermodynamic interaction with the prokaryotic cell membrane . On the basis of the molecular properties of Yoshixol, future biological interests and possible biological effects of Yoshixol are suggested.

Gen Pharmacol, 1997 May, 28(5), 733 - 5
Defibrotide augments the anticandidial activity of NK cells; Turhan A et al.; 1 . The mechanism of action of Defibrotide, a fibrinolytic agent on Natural Killer (NK) cell cytotoxicity, was investigated through verapamil, TMB-8 cells and pertussis toxin . 2 . Defibrotide increased the activity against Candida albicans cells (anticandidial activity), and it is determined that the calcium channels have a role in this effect . 3 . Blockage of calcium channels reduced the anticandidial effect by 39.2% . Pertussis toxin led to a 10.7% inhibition, whereas the application of TMB-8 resulted in the stimulation of anticandidial activity . 4 . It is concluded that defibrotide is a potent activator of NK cells.

Pharmazie, 1997 May, 52(5), 350 - 7
Synthesis of acyclo-C-nucleosides: 2-(alditol-1-yl)-5-methylthio- and -5-benzylthio-1,3,4-thiadiazoles; Shaban MA et al.; Condensation of S-methylhydrazinecarbodithioate or S-benzylhydrazinecarbodithioate with aldopentoses or aldohexoses gave the corresponding aldehydo-sugar S-methylhydrazonecarbodithioates of S-benzylhydrazonecarbodithioates . Oxidative cyclization of these hydrazones with bromine in acetic acid gave the corresponding 2-(alditol-1-yl)-5-alkylthio-1,3,4-thiadiazoles . Acetylation of the latter gave the corresponding per-O-acetyl derivatives which were also obtained by one-pot preparation by treatment of the hydrazones with bromine and sodium acetate in acetic acid followed by acetic anhydride . Some of the prepared compounds were tested for antimicrobial activity against Escherichia coli, Bacillus subtilis, Staphylococcus aureus and Candida albicans . While hydrazones showed significant activity against these organisms, the thiadiazoles were devoid of antimicrobial activity.

J Oral Rehabil, 1997 May, 24(5), 350 - 7
Antifungal effect of zeolite-incorporated tissue conditioner against Candida albicans growth and/or acid production; Nikawa H et al.; A new antimicrobial material, Ag-zeolite (Zeomic), was combined with a commercial tissue conditioner (GC-Soft Liner (GC); 1-5%) and, through monitoring the pH of the growth medium, examined for effects on the in vitro growth and/or acid production of Candida albicans on protein-free and saliva-coated specimens . The effect of incorporation of this agent on the physical property of the lining material was also examined according to the ISO penetration test . Comparison studies were carried out using GC, Coe Comfort (CC) or undecylenate combined GC (1-5%) specimens . Although the pH changes in the media varied depending upon the materials on which the Candida was grown, reverse sigmoidal pH curves were observed with most samples . As compared with GC, the soft lining materials showed, to some extent, an inhibitory effect on the acid production and/or the growth of C . albicans . These inhibitory effects consisted of a delay in the onset of rapid pH decline, decreases in the rate of pH change and increases in minimum pH . In most cases, the inhibitory effects of test specimens were dose-dependent, and zeolite specimens showed a significantly higher antifungal effect, followed by CC and undecylenate-combined GC; GC showed the least antifungal effect . The inhibitory effects of these materials on fungal growth were decreased by the presence of a saliva-coat, particularly with zeolite specimens and CC . However, four of eight 5%-Zeomic specimens still exhibited perfect growth inhibition in the presence of the salivary pellicle . Furthermore, test specimens containing 2-5% Zeomic showed a significantly greater effect on the delay in rapid decline of pH, as compared with the other specimens examined . In addition, the significantly higher minimum pH was observed where the yeasts were grown on 4%- and 5%-Zeomic specimens . The physical properties of all the test specimens conformed with the ISO standard as examined by penetration test . These results taken together suggest that an antimicrobial zeolite-combined tissue conditioner would be a potential aid in denture plaque control.

Clin Exp Allergy, 1997 May, 27(5), 584 - 92
Cell surface expression of two major yeast allergens in the Pityrosporum genus; Zargari A et al.; BACKGROUND: We have previously identified two major allergens of Pityrosporum orbiculare and characterized these as 37 kDa and 67 kDa proteins . OBJECTIVE: In the present study we have investigated the presence and subcellular location of the 37 kDa and 67 kDa allergen components in various members of the genus Pityrosporum as well as in Candida albicans, Candida parapsilosis and Saccharomyces cerevisiae . METHODS: To detect both cell surface and intracellular expression of the allergens, flow cytometry and confocal laser scanning microscopy (CLSM) were used . The cells were stained with indirect immunofluorescent (IIF) or alkaline phosphatase anti-alkaline phosphatase (APAAP) methods using mouse monoclonal antibodies (MoAbs) . RESULTS: Ninety-five per cent of the P . orbiculare (P . ovale) cells cultured for 4 days showed cell surface-binding of the anti-37 kDa MoAb and 88% of the cells bound the anti-67 kDa MoAb when analysed with IIF and flow cytometry . It was found that the members of the genus Pityrosporum (Malassezia), P . pachydermatis and M . sympodialis, expressed the 37 kDa and 67 kDa allergens to a similar extent as did P . orbiculare . Less than 5% of the cells of the genus Candida and S . cerevisiae showed positive staining with the MoAbs . The CLSM revealed that the 37 kDa and the 67 kDa components were located to the cell wall and could not be detected inside the acetone fixed and APAAP stained yeast cells of the genus Pityrosporum . When the yeast cells were cultured for more than 4 days the expression of both allergens decreased significantly . CONCLUSION: All three members of the genus Pityrosporum express the 37 kDa and 67 kDa major allergens on the cell surface, whereas these proteins could virtually not be detected in the Candida genus and S . cerevisiae.

J Oral Pathol Med, 1997 May, 26(5), 233 - 6
The in vitro adhesion of Candida albicans to buccal epithelial cells (BEC) from diabetic and non-diabetic individuals after in vivo and in vitro application of nystatin; Darwazeh AM et al.; Buccal epithelial cells (BEC) from 12 patients with diabetes mellitus and 12 age- and sex-matched non-diabetic subjects were tested in vitro for adhesion of Candida albicans following exposure to nystatin both in vitro and in vivo . Adhesion was significantly reduced (P < 0.002) to cells from both the diabetic and non-diabetic subjects after in vitro exposure to nystatin, but the reduction in adhesion was variable (5.0-50.7% in control subjects and 0.5-48.4% in diabetic subjects) and equivalent between the two groups . In vivo exposure to nystatin produced no overall significant reduction in candidal adhesion to cells from either diabetic or control subjects.

J Matern Fetal Med, 1997 May-Jun, 6(3), 151 - 4
Candida chorioamnionitis: a report of two cases; Berry DL et al.; Intra-amniotic infection (IAI) across intact membranes by Candida albicans is a rare occurrence . We report two cases of preterm labor and Candida chorioamnionitis that were diagnosed by intrapartum amniocentesis . The diverse maternal and neonatal outcomes observed indicate that Candida IAI is associated with significant, unpredictable maternal and neonatal morbidity.

Microbiology, 1997 May, 143 ( Pt 5), 1765 - 78
Variation in assimilating functions occurs in spontaneous Candida albicans mutants having chromosomal alterations; Rustchenko EP et al.; In this study, four clinical isolates and over 100 colony morphology mutants, previously derived spontaneously from strain 3153A during growth on glucose medium, were examined for their utilization of 21 carbon and 3 nitrogen sources at various growth temperatures . The results demonstrated extensive variability in the pattern of assimilation among the mutants and strains, including both the gain and loss of assimilating functions . The persistent alterations in assimilation patterns observed in sequentially produced subclones illustrated an extensive ability of C . albicans populations to constantly produce new combinations of assimilating functions . The variability among spontaneous mutants derived from a single strain explains the well documented variability among natural isolates . From these results we established a relationship between the previously documented broad spectrum of spontaneous chromosomal aberrations in these mutants to the expression of genes controlling the utilization of alternative carbon and nitrogen sources . The existence of cryptic genes, responsible for growth on alternative substrates, was previously deduced from the analysis of other mutants obtained as a response to the restrictive condition on media containing non-assimilating carbon sources . Thus, mutants with altered assimilation functions can arise either on glucose medium or by selection on restricted media . Extensive differences between the patterns of chromosomal aberrations and the distribution of correlated phenotypes in the two groups of mutants indicated that the same phenotypes may be produced by two different mechanisms involving the same or different genes.

Pharmacotherapy, 1997 May-Jun, 17(3), 538 - 48
The fluconazole era: management of hematogenously disseminated candidiasis in the nonneutropenic patient; Kramer KM et al.; Hematogenously disseminated candidiasis arising from nosocomial fungal infection is a life-threatening complication in critically ill, nonneutropenic patients . The overall nosocomial fungal infection rate in United States hospitals doubled from 1980-1990 . Until recently, amphotericin B was the only agent available for the treatment of life-threatening candidal infections, but its use is plagued by toxicities including nephrotoxicity and infusion-related reactions such as rigors and hypotension . The availability of fluconazole, which is regarded as much less toxic than amphotericin B, prompted a surge in research to determine if it is as efficacious in the management of candidemia and hematogenously disseminated candidiasis . Complicating the interpretation of studies is the broad range of infection severity, from candidemia that may be transient and self-limiting to life-threatening hematogenously disseminated candidiasis . Clinical trials comparing fluconazole and amphotericin B demonstrate the efficacy of fluconazole for catheter-associated candidemia in critically ill patients when the likely pathogen is Candida albicans . Amphotericin B should remain the first-line agent for the management of candidemia and hematogenously disseminated candidiasis in all other patients.

J Prosthet Dent, 1997 May, 77(5), 535 - 9
Retention of Candida albicans on acrylic resin and silicone of different surface topography; Verran J et al.; STATEMENT OF PROBLEM: The adhesion of microorganisms to a denture surface is a prerequisite for colonization . PURPOSE: This study compared the retention of Candida albicans on smooth and rough acrylic resin and silicone surfaces after a washing procedure to determine the effect of surface roughness on prosthesis infection and hygiene . MATERIAL AND METHODS: Standardized cell suspensions of C . albicans were incubated with smooth and rough acrylic resin and silicone surfaces for 1 hour at 24 degrees C . After washing, cells that had been retained on the surface were stained with acridine orange and examined with incident beam fluorescent microscopy . RESULTS: There was no significant difference in cell numbers on either of the smooth surfaces . Significantly higher numbers of cells (p > 0.0005) were observed on roughened surfaces (silicone > acrylic resin) than on smooth surfaces . The fitting surface of the maxillary denture was not polished . CONCLUSIONS: Silicones used in prostheses were processed against dental stone . The resultant surface roughness may facilitate microbial retention and infection and should therefore be kept to a minimum.

Antimicrob Agents Chemother, 1997 May, 41(5), 1156 - 7
In vitro activity of a new echinocandin, LY303366, compared with those of amphotericin B and fluconazole against clinical yeast isolates; Uzun O et al.; The in vitro activity of LY303366, a new echinocandin derivative, was evaluated with 191 yeast isolates by a broth microdilution method . The MICs at which 50% of the isolates were inhibited were 0.125 microg/ml for Candida albicans and C . tropicalis, 0.25 microg/ml for C . krusei, C . kefyr, and C . glabrata, and 2.0 microg/ml for C . parapsilosis.

Clin Diagn Lab Immunol, 1997 May, 4(3), 328 - 33
Differential humoral response against alpha- and beta-linked mannose residues associated with tissue invasion by Candida albicans; Jouault T et al.; Candida albicans mannan is the major cell wall antigen that elicits antibodies considered to be of little diagnostic value . It comprises epitopes corresponding to sequences of alpha- and beta-1,2-linked mannose residues . Both types of oligomannosidic epitopes may also be present on the glycosidic portions of other C . albicans molecules, i.e., mannoproteins (MP) (either structural or enzymatic) and glycolipids . The human humoral responses against beta-1,2- and alpha-linked oligomannosides were investigated by C . albicans Western blotting by considering the elective distribution of beta-1,2-oligomannosidic epitopes over a 14- to 18-kDa phospholipomannan (PLM) and the presence of alpha-mannosidic epitopes over heavily glycosylated MP . Western blotting of 51 control sera confirmed the presence of antibodies against C . albicans as a commensal member of the indigenous microflora; an immunoglobulin G (IgG) reactivity linked to enzyme-linked immunosorbent assay mannan signals was found for both PLM (beta-1,2-Man residues) and MP (alpha-Man residues) . Despite strong reactivities against mannan and MP, IgG from 21 hospitalized patients with mycological evidence of deep-tissue invasion by C . albicans very significantly failed to react or reacted only faintly with PLM . This downregulation of anti-beta-1,2-oligomannosidic epitopes, associated with tissue invasion by C . albicans, was confirmed in 3 of 4 AIDS patients with extended oroesophageal candidosis . The application of a dissociation procedure proved that the absence of PLM reactivity was not due to the presence of immune complexes . These data provide the first evidence for a qualitative modification of the human antimannan antibody response associated with the C . albicans commensal-pathogenic transition.

AIDS, 1997 May, 11(6), 759 - 63
A dose comparison study of a new triazole antifungal (D0870) in HIV-positive patients with oral candidiasis; De Wit S et al.; OBJECTIVE: This multicentre study evaluated the clinical efficacy and tolerability of D0870 in treating oropharyngeal candidiasis in HIV-positive patients who had no history of clinical resistance to fluconazole . METHODS: Three regimens were evaluated in two phases . In phase I a 50 mg initial dose was followed by 10 mg for 4 days (Group 1) . In phase II a 100 mg initial dose was followed by 25 mg for 4 days (Group 2), or 10 mg for 5 days (Group 3) . RESULTS: Clinical cure was obtained in 27 patients of a total of 35 (77%) and six other patients improved (17%) . Two patients at the lowest dose failed and both had very low plasma concentration of D0870 . No association was found between clinical outcome; minimum inhibitory concentration of D0870 pre-therapy for Candida albicans, maximum recorded plasma D0870 concentration, cfu of culture or CD4 cell count at entry . Overall, 37% of the patients experienced relapse during the 2 weeks post therapy . Tolerance was excellent . Mild adverse events possibly related to the study drug were recorded in five patients . CONCLUSION: D0870 demonstrates excellent efficacy at low doses in the treatment of HIV-related OPC and exhibits a favourable safety profile.

Chemotherapy, 1997 May-Jun, 43(3), 198 - 203
Antifungal and immunoadjuvant properties of fluconazole in mice immunosuppressed with morphine; Di Francesco P et al.; We investigated the efficacy of fluconazole on experimental disseminated candidiasis in mice immunocompromised by chronic morphine treatment . CD1 mice were severely immunosuppressed by repeated morphine administrations, i.e., subcutaneous (s.c.) injections of 75 mg/kg/day, 3 days before and 5 days after a systemic Candida albicans infection induced by intravenous administration of 1 x 10(6) fungal cells/mouse . Fluconazole (2.5 mg/kg, s.c., at 6, 24 and 48 h postinfection) was very effective in prolonging survival time of morphine-treated mice . Fluconazole treatment also promotes a recovery of killing activity of polymorphonuclear leukocyte cells suppressed by morphine administrations.

Ann Intern Med, 1997 May 1, 126(9), 689 - 96
Weekly fluconazole for the prevention of mucosal candidiasis in women with HIV infection . A randomized, double-blind, placebo-controlled trial . Terry Beirn Community Programs for Clinical Research on AIDS; Schuman P et al.; BACKGROUND: Candidiasis is a frequent complication of infection with the human immunodeficiency virus (HIV); however, few data exist about the natural history, prevention, and treatment of mucosal candidiasis in women . OBJECTIVE: To evaluate the safety and effectiveness of weekly fluconazole prophylaxis for mucosal candidiasis in women infected with HIV . DESIGN: Randomized, double-blind, placebo-controlled trial . SETTING: 14 sites participating in the Community Programs for Clinical Research on AIDS (CPCRA) . PATIENTS: 323 women with HIV infection and CD4+ cell counts of 300 cells/mm3 or less . INTERVENTION: 200 mg of fluconazole per week or placebo . Open-label fluconazole for candidiasis prophylaxis was permitted after two oropharyngeal or vaginal episodes or one esophageal episode . MEASUREMENTS: Development of mucosal candidiasis, clinical and in vitro resistance of Candida species to fluconazole, survival, and adverse events . RESULTS: After a median follow-up of 29 months, 72 of 162 patients receiving fluconazole and 93 of 161 patients receiving placebo had at least one episode of candidiasis (relative risk {RR}, 0.56 {95% Cl, 0.41 to 0.77); P < 0.001) . Weekly fluconazole was effective in preventing oropharyngeal candidiasis (RR, 0.50 {Cl, 0.33 to 0.74}; P < 0.001) and vaginal candidiasis (RR, 0.64 {Cl, 0.40 to 1.00}; P = 0.05) but not esophageal candidiasis (RR, 0.91 {Cl, 0.48 to 1.72}; P > 0.2) . Relative risks were similar for women who had a history of mucosal candidiasis (RR, 0.5 {Cl, 0.35 to 0.75}) and those who did not (RR, 0.69 {Cl, 0.35 to 1.34}) . Absolute risk reduction for patients with a history of infection was 25.6 per 100 person-years, which is more than twice the reduction of 11.2 per 100 person-years seen in patients with no history of infection . This difference reflects the higher risk of patients who previously had an infection . Candida albicans was not usually resistant to fluconazole in vaginal specimens in clinical or in vitro settings; such resistance occurred in less than 5% of patients in each group . CONCLUSIONS: Weekly fluconazole (200 mg) seems to be safe and effective in preventing oropharyngeal and vaginal candidiasis . This regimen has a useful role in the management of HIV-infected women who are at risk for recurrent mucosal candidiasis.

J Infect Dis, 1997 May, 175(5), 1169 - 75
Assessment of a mouse model of neutropenia and the effect of an anti-candidiasis monoclonal antibody in these animals; Han Y et al.; As previously reported, monoclonal antibody (MAb) B6.1 increases resistance to hematogenous disseminated candidiasis in normal and SCID mice . In this study, MAb B6.1 was examined in a mouse model of neutropenia . The neutropenia was induced for a short period of time by a single dose of the anti-neutrophil antibody, MAb RB6-8C5, or for a protracted period by doses of MAb RB6-8C5 every other day . At low doses (< or = 25 microg/mouse), neutrophils were primarily affected, but at high doses (> or = 50 microg/mouse), lymphocytes were also depleted . Mice given either single or multiple doses of MAb RB6-8C5 were more resistant to experimental hematogenous disseminated candidiasis if they received MAb B6.1 before and after challenge with Candida albicans yeast cells intravenously . These results show the utility of MAb RB6-8C5 for induction of a protracted neutropenia in mice and demonstrate that MAb B6.1 can enhance resistance against candidiasis under neutropenic conditions.

J Immunol, 1997 May 1, 158(9), 4328 - 35
B cell-independent selection of memory T cells after mucosal immunization with Candida albicans; Jones-Carson J et al.; B cell-deficient mice have normal T cell responses to Ags inoculated systemically; however, it is not known whether they can mount systemic and mucosal T cell responses to Ags through normally B cell-enriched gastrointestinal mucosae . Mucosal colonization of germfree, B cell-deficient J(H)D mice with the pathogenic gastrointestinal fungus, Candida albicans selected splenic CD4+ and CD8+ TCR alphabeta memory T cells, as indicated by 1) increased numbers of splenic CD4+ and CD8+ TCR alphabeta expressing T cells of the CD45RB(low) CD44(high) phenotype, 2) early expansion followed by progressive decrease in the number of splenic CD4+ and CD8+ TCR alphabeta T cells, and 3) concomitant increases in the percentage of apoptosis and proliferation in the latter subsets . Although i.v . challenge of germfree or conventional J(H)D mice with C . albicans did not increase apoptosis or induce changes in the number of splenic memory T cells, i.v . challenge of mucosally immunized germfree J(H)D mice led to further proliferation and expansion of activated splenic CD4+ and CD8+ TCR alphabeta thymic-educated memory T cells, which were first evoked by mucosal immunization . Oral colonization with C . albicans also increased the number of gammadelta and thymic and extrathymic alphabeta T cells in gastrointestinal mucosae . In conclusion, our results are the first strong evidence that thymic and extrathymic T cells participate in mucosal immunity to C . albicans in the absence of B cells; however, CD4+ and thymic-educated CD8+ TCR alphabeta memory subsets evoked by mucosal, but not parenteral (i.v.), challenge contribute to protective immunity to systemic candidiasis.

Infect Immun, 1997 May, 65(5), 1748 - 53
Gamma interferon is not essential in host defense against disseminated candidiasis in mice; Qian Q et al.; In vitro studies have suggested a role for interferon gamma (IFN-gamma) in host defense against disseminated candidiasis, but in vivo studies are inconclusive . We utilized homozygous IFN-gamma knockout (GKO) mice to determine if the cytokine is essential in host defense against this disease . Genotypes of mice were determined by PCR with specific primers for the normal or disrupted IFN-gamma gene . The GKO status of the mice was confirmed by an enzyme-linked immunosorbent assay, which showed no detectable IFN-gamma produced by their splenocytes stimulated by concanavalin A . To test the susceptibility of GKO mice to candidiasis, the animals were infected either intravenously (i.v.) or intragastrically (i.g.) with Candida albicans . GKO mice infected i.v . survived as long as wild-type (WT) mice and showed no difference in Candida CFU counts in liver, spleen, or kidneys compared to those for WT mice . When animals were given Candida i.g., at 3 h or at 10 or 21 days after infection, there was no dissemination of Candida to the lung, liver, spleen, or kidneys for either GKO or WT mice . There was no difference in Candida CFU counts recovered from the stomach or intestines between GKO and WT mice . Histological examination of the stomach cardial-atrium fold, where the fungus was located, showed that GKO mice did not have evidence of more tissue damage or fungal invasion than WT mice . Finally, the jejunum for both types of mice showed no evidence of tissue damage or fungal invasion . These studies indicate that IFN-gamma is not essential in host defense against C . albicans that originates from a mucosal site or that is given directly into the bloodstream in a mouse model.

J Clin Microbiol, 1997 May, 35(5), 1263 - 5
Evaluation of bichro-latex albicans, a new method for rapid identification of Candida albicans; Quindos G et al.; A method for identification of Candida albicans within 5 min was evaluated by using 4,643 yeast isolates . Six false-positive and three false-negative reactions were observed . The specificity (99.87%) and sensitivity (99.74%) obtained indicate that the Bichro-latex albicans test is a useful method for the rapid identification of C . albicans colonies.

Mol Cells, 1997 Apr 30, 7(2), 214 - 9
Purification and characterization of an antifungal PR-5 protein from pumpkin leaves; Cheong NE et al.; A 28-kDa antifungal PR-5 protein (PLTP) was purified from pumpkin leaves to homogeneity by using ammonium sulfate fractionation, a regenerated chitin column, and reversed-phase column chromatographies on butyl-Toyopearl and HPLC C18 columns . Analysis of 14 N-terminal amino acid sequences of PLTP shows 100% sequence identity to those of two PR-5 proteins, NP24 from tomatoes and AP24 from tobacco . The identical sequence also exhibited high amino acid sequence homology to that of an osmotin-like protein (OLP; 71%) from tobacco cells and thaumatin (64%), a sweet-tasting protein of Thaumatococcus danielli Bench . When the PLTP was immuno-blotted with antiserum raised against the tobacco OLP, the OLP antibody specifically cross-reacted with the PLTP, suggesting that they share several common epitopes in their tertiary structure of the proteins . The purified PLTP rapidly lyzed hyphal tips of Neurospora crassa at a concentration greater than 200 nM and significantly inhibited the fungal growth of Fusarium oxysporum in an agar-disc plate at a concentration greater than 2 microM . It also shows a synergistic effect with nikkomycin, a chitin synthase inhibitor, for the growth inhibition of Candida albicans.

Gene, 1997 Apr 29, 190(1), 99 - 104
A novel MAP-kinase kinase from Candida albicans; Singh P et al.; A putative MAP-kinase kinase-encoding gene, CaSTE7, was isolated from Candida albicans by complementation of ste7 and ste11 mutants of the pheromone signal-transduction pathway of Saccharomyces cerevisiae . The nucleotide (nt) sequence revealed an ORF of 1767 nt encoding a putative protein of 589 amino acids (aa) . CaSTE7 has a strong homology with MAP-kinase kinase STE7 of S . cerevisiae, the kinase domain having 45% homology with that of STE7 . The deduced aa sequence contained all eleven consensus kinase subdomains found in MAP-kinase kinases . It can suppress the mating defect of ste5, ste11, ste7, and fus3 kss1 double mutants, but it cannot bypass the ste12 mutation . CaSTE7 behaves as a hyperactive allele of STE7, suppressing the mating defects of the pheromone signal-transduction pathway by constitutively stimulating STE12, and hence STE12-dependent processes.

Mol Gen Genet, 1997 Apr 28, 254(4), 417 - 26
Multidrug resistance in Aspergillus nidulans involves novel ATP-binding cassette transporters; Del Sorbo G et al.; Two single-copy genes, designated atrA and atrB (ATP-binding cassette transporter A and B), were cloned from the filamentous fungus Aspergillus nidulans and sequenced . Based on the presence of conserved motifs and on hydropathy analysis, the products encoded by atrA and atrB can be regarded as novel members of the ATP-binding cassette (ABC) superfamily of membrane transporters . Both products share the same topology as the ABC transporters PDR5 and SNQ2 from Saccharomyces cerevisiae and CDR1 from Candida albicans, which are involved in multidrug resistance of these yeasts . Significant homology also occurs between the ATP-binding cassettes of AtrA and AtrB, and those of mammalian ABC transporters (P-glycoproteins) . The transcription of atrA and, in particular, atrB in mycelium of A . nidulans is strongly enhanced by treatment with several drugs, including antibiotics, azole fungicides and plant defense toxins . The enhanced transcription is detectable within a few minutes after drug treatment and coincides with the beginning of energy-dependent drug efflux activity, reported previously in the fungus for azole fungicides . Transcription of the atr genes has been studied in a wild-type and in a series of isogenic strains carrying the imaA and/or imaB genes, which confer multidrug resistance to various toxic compounds such as the azole fungicide imazalil . atrB is constitutively transcribed at a low level in the wild-type and in strains carrying imaA or imaB . Imazalil treatment enhances transcription of atrB to a similar extent in all strains tested . atrA, unlike atrB, displays a relatively high level of constitutive expression in mutants carrying imaB . Imazalil enhances transcription of atrA more strongly in imaB mutants, suggesting that the imaB locus regulates atrA . Functional analysis demonstrated that cDNA of atrB can complement the drug hypersensitivity associated with PDR5 deficiency in S . cerevisiae.

EMBO J, 1997 Apr 15, 16(8), 1982 - 91
Efg1p, an essential regulator of morphogenesis of the human pathogen Candida albicans, is a member of a conserved class of bHLH proteins regulating morphogenetic processes in fungi; Stoldt VR et al.; We identified a gene of the fungal pathogen Candida albicans, designated EFG1, whose high-level expression stimulates pseudohyphal morphogenesis in the yeast Saccharomyces cerevisiae . In a central region the deduced Efg1 protein is highly homologous to the StuA and Phd1/Sok2 proteins that regulate morphogenesis of Aspergillus nidulans and S . cerevisiae, respectively . The core of the conserved region is homologous to the basic helix-loop-helix (bHLH) motif of eukaryotic transcription factors, specifically to the human Myc and Max proteins . Fungal-specific residues in the bHLH domain include the substitution of an invariant glutamate, responsible for target (E-box) specificity, by a threonine residue . During hyphal induction EFG1 transcript levels decline to low levels; downregulation is effected at the level of transcriptional initiation as shown by a EFG1 promoter-LAC4 fusion . A strain carrying one disrupted EFG1 allele and one EFG1 allele under the control of the glucose-repressible PCK1 promoter forms rod-like, pseudohyphal cells, but is unable to form true hyphae on glucose-containing media . Overexpression of EFG1 in C . albicans leads to enhanced filamentous growth in the form of extended pseudohyphae in liquid and on solid media . The results suggest that Efg1p has a dual role as a transcriptional activator and repressor, whose balanced activity is essential for yeast, pseudohyphal and hyphal morphogenesis of C . albicans . Functional analogies between Efg1p and Myc are discussed.

Arch Biochem Biophys, 1997 Apr 15, 340(2), 347 - 54
Purification and characterization of protein kinase CK2 from Candida albicans: evidence for the presence of two distinct regulatory subunits beta and beta'; Walz K et al.; Protein kinase CK2 of Candida albicans has been purified to near homogeneity by a procedure which involves chromatography on DEAE-cellulose, phosphocellulose, Q-Sepharose, and heparin-agarose . The purified enzyme has the characteristic properties of animal and yeast CK2, i.e., it utilizes ATP as well as GTP as phosphate donor, phosphorylates serine and threonine residues on casein, is inhibited by low concentrations of heparin, and is stimulated by NaCl and polycationic compounds such as polylysine, spermine, and spermidine . The native form of the enzyme exhibits a molecular mass of 159 kDa, and SDS-PAGE analysis indicates that it is composed of four polypeptides with relative molecular masses of 44, 39, 37 and 36 kDa . The 39- and 37-kDa polypeptides were identified as distinct catalytic subunits alpha and alpha' on the basis of in situ phosphorylation assays and immunological recognition with heterologous antibodies . The purified kinase undergoes autophosphorylation on the 44- and 36-kDa polypeptides, a characteristic of the beta subunits from other species . Antibodies raised against the beta subunit of Drosophila melanogaster and human CK2 crossreact only with the 36-kDa polypeptide . The 44-kDa polypeptide was identified as an unusually large beta' subunit by Western blotting with an antibody raised against the beta' subunit of Saccharomyces cerevisiae . All these data suggest that C . albicans CK2 has an alpha alpha' beta beta' heterotetrameric composition similar to that found in S . cerevisiae.

Arch Biochem Biophys, 1997 Apr 15, 340(2), 265 - 9
Differential inhibition of fungal amd mammalian squalene epoxidases by the benzylamine SDZ SBA 586 in comparison with the allylamine terbinafine; Favre B et al.; The allylamine class of antifungal compounds are specific inhibitors of squalene epoxidase (SE) . However, depending on their chemical structure, allylamine derivatives can be highly selective for either fungal or mammalian SEs . All allylamines tested previously, irrespective of their selectivity, inhibit fungal SEs in a noncompetitive manner and mammalian SEs in a competitive manner . Here we have analyzed the inhibitory properties of the benzylamine SDZ SBA 586 toward fungal and mammalian SEs in comparison to the systemic antimycotic terbinafine, SDZ SBA 586 was, like terbinafine a selective inhibitor of fungal SE . Microsomal SE from the pathogenic yeast candida albicans was sixfold more sensitive to SDZ SBA 586 than to terbinafine, C50: 8 nM versus 44 nM, while the enzyme from the dermatophyte fungus Trichophyton rubrum was slightly less sensitive to SDZ SBA 586 than to terbinafine, IC50: 39 and 18 nM, respectively . Similarly to terbinafine, SDZ SBA 586 inhibited the yeast enzyme in non competitive manner, SDZ SBA 586 also inhibited mammalian microsomal SEs, but only at micromolar concentrations . It was more active than terbinafine toward both guinea pig SE, IC50: 2 microM versus 4 microM, and rat SE, IC50: 11 microM versus 87 microM . However, in contrast to terbinafine as well as allylamines selective for mammalian SE, SDZ SBA 586 was a noncompetitive inhibitor of rat microsomal SE . Interestingly, depending on the source of microsomal SE, binding of terbinafine and SDZ SBA 586 exhibited a positive, indifferent, or negative cooperativity, suggesting that SE is an oligomeric enzyme.

FEMS Microbiol Lett, 1997 Apr 15, 149(2), 141 - 9
Molecular mechanisms of azole resistance in fungi; Joseph-Horne T et al.; This paper reviews the current status of our understanding of azole antifungal resistance mechanisms at the molecular level and explores their implications . Extensive biochemical studies have highlighted a significant diversity in mechanisms conferring resistance to azoles, which include alterations in sterol biosynthesis, target site, uptake and efflux . In stark contrast, few examples document the molecular basis of azole resistance . Those that do refer almost exclusively to mechanisms in laboratory mutants, with the exception of the role of multi-drug resistance proteins in clinical isolates of Candida albicans . It is clear that the technologies required to examine and define azole resistance mechanisms at the molecular level exist, but research appears distinctly lacking in this most important area.

Gene, 1997 Apr 11, 189(1), 119 - 26
Cloning and expression of squalene epoxidase from the pathogenic yeast Candida albicans; Favre B et al.; The allylamine antimycotic terbinafine prevents the formation of sterols by specifically inhibiting squalene epoxidase (SE) . The biological and biochemical action of terbinafine on fungal pathogens has been well investigated, but little is known at the molecular level . Here we report the cloning, sequencing and expression of the target of terbinafine from the major pathogen Candida albicans . A C . albicans genomic DNA library was constructed in gamma ZAP Express and screened with a DNA fragment obtained by polymerase chain reaction with two primers designed from sequences common to Saccharomyces cerevisiae and rodent SEs . Two types of clone, approximately 3.9 kbp and 4.1 kbp, were isolated . Both contained an identical open reading frame of 1488 nucleotides, while a few sequence differences were found in the flanking regions, suggesting an allelic heterogeneity . The deduced protein sequence of C . albicans SE, 496 amino acids (55324 Da), is 54% and 41% identical to those of S . cerevisiae and rat, respectively . A 1.8-kb transcript was observed on Northern blots of C . albicans mRNA . Polyclonal antibodies, raised against an internal peptide of C . albicans SE, recognized a protein associated with the particulate fraction of M(r) 55000 on Western blots of C . albicans extracts . C . albicans SE was overexpressed in S . cerevisiae with the expression vector pYES2 . In homogenates from S . cerevisiae overexpressing the C . albicans protein SE activity was 10-fold higher than the endogenous activity from controls.

AIDS Res Hum Retroviruses, 1997 Apr 10, 13(6), 485 - 91
Molecular epidemiology of mucosal candidiasis in HIV-positive women; Dahl KM et al.; Mucosal candidiasis is a common complication of HIV infection and HIV-positive women may develop both oropharyngeal and vaginal disease . Colonization with Candida albicans and related species at either site is a common preceding event in asymptomatic women . To examine the molecular epidemiology of colonizing yeast strains in HIV-positive women, concurrent oropharyngeal and vaginal cultures were obtained from 32 women (mean CD4 count 392 cells/mm3, range 0-1319) . Positive oropharyngeal cultures were obtained in 18 (56%) and positive vaginal cultures in 10 (31%) . Candida species were isolated from both sites simultaneously in nine (28%) women . All strains were evaluated for restriction fragment length polymorphisms (RFLPs) at the ribosomal DNA locus (using a heterologous 8.4-kb NotI probe from H . capsulatum) and with a C . albicans-specific repetitive DNA probe . Isolates were grouped into three classes by the NotI probe and then members of each class were evaluated with the C . albicans-specific probe . Isolates were subsequently evaluated by random amplified polymorphic DNA (RAPD) PCR with four arbitrary primers to detect strain-specific differences . All isolates tested were unique and could be discriminated by RFLP or RAPD PCR . Vaginal and oropharyngeal isolates from the same individual in all nine cases were dissimilar, suggesting that the dominant strain of Candida colonizing different body sites is different . These findings suggest that the epidemiology of Candida infection in HIV disease is complex, that the development of oropharyngeal and vaginal disease may be disassociated, and that HIV-positive patients are each infected by their own unique strains of Candida.

Oral Microbiol Immunol, 1997 Apr, 12(2), 126 - 8
In vitro antifungal resistance of oral Candida albicans strains in non-AIDS patients; Monteil RA et al.; Some cases of oral candidosis are refractory to antifungal treatment . This might be related to development of resistant Candida strains, but susceptibility testing is not standardized and not routinely available, and information related to this problem is scarce in non-AIDS patients . In this study, the in vitro antifungal resistance of oral Candida albicans strains was evaluated . The strains were obtained from a cohort of 72 HIV-negative patients with oral yeast carriage and clinical complaint . Laboratory identification revealed C . albicans in 93% of cases . None of these oral C . albicans isolates showed in vitro resistance to polyenes, but they showed varying resistance levels to fluorocytosine and azoles . This study confirms the usefulness of standardizing susceptibility testing so that it could be routinely available and of realizing a mycological diagnosis including an antifungigram when oral candidosis is suspected, whenever antifungal treatment with azoles is planned.

Allergy, 1997 Apr, 52(4), 394 - 403
Anti-Candida albicans IgE and IgG subclasses in sera of patients with allergic bronchopulmonary aspergillosis (ABPA); Roig E et al.; We performed immunoblotting experiments to determine specific IgE and IgG subclass responses to Candida albicans antigens in allergic bronchopulmonary aspergillosis (ABPA) patients . This is a first report describing C . albicans antigens recognized by serum IgE and IgG subclasses of ABPA patients sensitized to that yeast . Among the various antigens reacting with serum IgE, a 43-kDa component was recognized by all seven patients and can be considered a major antigen of C . albicans for this particular group of patients . By comparison, only 20% of a group of asthmatic atopics (25 patients) and 10% of a group of normal controls (10 subjects) were 43-kDa positive . Multiple banding patterns, revealing no major antigen, were observed for all four IgG subclasses except for IgG1 in one case . In particular, the 43-kDa component was not always recognized by all the patients . Furthermore, oral or inhaled steroid treatment appears to have no impact on the specific IgE immunopatterns obtained . Using immunoelectron-microscopy, we localized IgE-binding primarily in the mannoprotein-containing layers of the C . albicans cell wall . In conclusion, C . albicans-IgE and IgG subclasses may participate in the physiopathology of ABPA by exacerbating pulmonary infiltrates (IgE) and inducing eosinophil-mediated inflammatory reaction (IgG1, IgG3).

Inflammation, 1997 Apr, 21(2), 159 - 72
Macrophage-mediated candidacidal activity is augmented by exposure to eosinophil peroxidase: a paradigm for eosinophil-macrophage interaction; Lefkowitz DL et al.; Various disease states are associated with eosinophilia and the release of eosinophil peroxidase (EPO) into the microenvironment . The present study targets the effects of low levels of EPO on macrophage (M phi) phagocytosis and intracellular killing of Candida albicans as well as M phi oxidative activity measured as the luminescence product of luminol dioxygenation . Resident murine peritoneal M phi were exposed to various concentrations of EPO . Chemiluminescence data indicate that nanomolar concentrations of EPO markedly enhanced the dioxygenation activity (respiratory burst) of M phi . In other studies, the exposure of M phi to 0.17 microM EPO for 10 min . enhanced M phi-mediated candidacidal activity 10 fold . The above data indicate that EPO enhances certain M phi functions . Also the results illustrate a previously un-recognized interaction between eosinophils and M phi and implicate yet another possible role for EPO in host defenses against disease.

Arch Pediatr, 1997 Apr, 4(4), 331 - 4
{Maternofetal disseminated candidiasis and high-grade prematurity}; Baud O et al.; BACKGROUND: Despite the frequency of vaginal yeast colonization, serious candidiasis infections in pregnant patients or neonates remain rare . Four cases of disseminated congenital candidiasis in very preterm infants are reported . CASE REPORTS: Congenital Candida albicans infection has been diagnosed in four very preterm infants . In three cases, the mothers had intrauterine devices in place throughout pregnancy . A careful macroscopic examination of the umbilical cord and placenta after birth has allowed an early management strategy in three cases . In all cases, a serious infectious alveolitis occurred . A pronounced increase in white blood cells (> 50,000/mm3) and high levels of both segmented neutrophil and band cells, despite the frequent normality of the CRP, constituted other features . Infection was controlled by parenteral amphotericin B or fluconazole . In one case, serious thrombocytopenia occurred after the first amphotericin B injection requiring substitution for fluconazole . The outcome was unfavourable in two cases with an extensive periventricular leukomalacia . CONCLUSION: Congenital candidiasis in these four very preterm neonates has several features in common: intrauterine contraceptive device during pregnancy, characteristic chorioamnionitis and funisitis, high WBC count, infectious alveolitis . Fluconazole as alternative to amphotericine B therapy is proposed.

J Clin Gastroenterol, 1997 Apr, 24(3), 165 - 8
Fatal diffuse invasive gastrointestinal candidiasis masking as ileus after bone marrow transplantation; DiBaise JK et al.; High-dose cytotoxic chemotherapy has increased the incidence of candidal infections that make neutropenic patients very sick and may kill them . We report fatal invasive candidiasis involving the entire alimentary tract after autologous bone marrow transplantation in a young woman with breast cancer . Illustrated are the significance of fungal infections in this patient population, the potential for Candida albicans to invade the entire gastrointestinal tract, and the potential role of endoscopy in the early diagnosis of these often catastrophic infections . We also suggest that diffuse, invasive candidiasis should be considered in the differential diagnosis of ileus in the immunocompromised patient.

Eur J Clin Microbiol Infect Dis, 1997 Apr, 16(4), 296 - 300
Use of specialised isolation media for recognition and identification of Candida dubliniensis isolates from HIV-infected patients; Schoofs A et al.; During a study of oral rinses of 130 HIV-infected individuals, both typical and atypical Candida albicans colonies were isolated from ten patients on a yeast differential medium . Typical Candida albicans colonies were light green; atypical colonies were dark green . Both types of colonies were germ tube-positive and produced chlamydospores . However, DNA fingerprinting of the atypical isolates with the Ca3 Candida albicans-specific probe showed that they belonged to the recently described species Candida dubliniensis . Candida dubliniensis colonies could also be differentiated from Candida albicans colonies on isolation plates by the absence of fluorescence of colonies on methyl blue-Sabouraud agar under Wood's light . Among other phenotypic characteristics, only the absence of intracellular beta-glucosidase activity reliably distinguished Candida albicans from Candida dubliniensis . Candida dubliniensis may be underreported in clinical samples because most currently used isolation and identification methods fail to recognize this yeast.

J Clin Microbiol, 1997 Apr, 35(4), 960 - 4
Widespread geographic distribution of oral Candida dubliniensis strains in human immunodeficiency virus-infected individuals; Sullivan D et al.; Candida dubliniensis is a recently identified chlamydospore-positive yeast species associated with oral candidiasis in human immunodeficiency virus (HIV)-infected (HIV+) patients and is closely related to Candida albicans . Several recent reports have described atypical oral Candida isolates with phenotypic and genetic properties similar to those of C . dubliniensis . In this study 10 atypical chlamydospore-positive oral isolates from HIV+ patients in Switzerland, the United Kingdom, and Argentina and 1 isolate from an HIV-negative Irish subject were compared to reference strains of C . albicans and Candida stellatoidea and reference strains of C . dubliniensis recovered from Irish and Australian HIV+ individuals . All 11 isolates were phenotypically and genetically similar to and phylogenetically identical to C . dubliniensis . These findings demonstrate that the geographical distribution of C . dubliniensis is widespread, and it is likely that it is a significant constituent of the normal oral flora with the potential to cause oral candidiasis, particularly in immunocompromised patients.

J Clin Microbiol, 1997 Apr, 35(4), 856 - 61
Comparison of four molecular typing methods for evaluating genetic diversity among Candida albicans isolates from human immunodeficiency virus-positive patients with oral candidiasis; Diaz-Guerra TM et al.; Candida albicans strain delineation by karyotyping . NotI restriction pattern analysis, hybridization with specific probe 27A, and PCR fingerprinting with the phage M13 core sequence were performed with 30 isolates from the oral cavities of 30 human immunodeficiency virus (HIV)-infected patients and 8 reference strains . Within the panel of clinical isolates, 20 were geographically related, although 10 isolates were susceptible to fluconazole and 10 isolates were resistant to fluconazole . The remaining isolates used in this study were fluconazole resistant and geographically unrelated . A composite DNA type was defined for each of the strains as the combination of types obtained by the four molecular methods . By this procedure, a great diversity of DNA types was found among isolates from the oropharynges of HIV-infected individuals with oral candidiasis . This diversity was not reduced when isolates were evaluated on the basis of whether they came from the same geographical locale and whether they were fluconazole resistant . These data refute the idea of a clonal origin for fluconazole-resistant strains among HIV-positive patients . Karyotyping was the least discriminatory method, yielding 19 DNA types among the 38 strains analyzed . Conversely, hybridization with the 27A probe showed a unique DNA pattern for each of the strains examined in this study . Our results demonstrate that at least two different molecular methods are needed for Candida albicans typing and that there is a great deal of strain variation within the species, irrespective of place of origin or antifungal resistance patterns.

J Vet Med Sci, 1997 Apr, 59(4), 271 - 6
Relationship between age-dependent changes of bovine neutrophil functions and their intracellular Ca2+ concentrations; Higuchi H et al.; Neutrophil functions and intracellular Ca2+ concentrations ({Ca2+}i) were evaluated in 15 Holstein cattle divided into the following 3 groups: 5 neonatal calves less than 1 week old (group 1), 5 young calves 2 to 4 weeks old (group 2) and 5 cows 2 to 3 years old (group 3) . The ability of neutrophils to phagocytose Candida albicans (C . albicans) was significantly higher (p < 0.05) in neonatal and young calves than in cows, whereas the phagocytosis by neutrophils of bovine IgG-coated yeasts (IgG-yeasts) was significantly lower (p < 0.05) in neonatal and young calves than that in cows . The killing activity by neutrophils of C . albicans in neonatal and young calves was significantly lower (p < 0.05) than that in cows . Luminol dependent chemiluminescent (LDCL) responses stimulated with opsonized zymosan (OPZ), heat-aggregated IgG (H-agg.IgG) and phorbol myristate acetate (PMA) were apparently lower in neonatal and young calves than in cows . No clearly different expressions of complement receptor type 3 (CR3) on neutrophils were observed among the 3 groups of cattle, although the values due to the binding of FITC-anti-bovine IgG to neutrophils in neonatal and young calves were lower than those in group 3 . The OPZ-induced {Ca2+}i of neutrophils in neonatal and young calves were significantly higher (p < 0.05) than those in cows, but they were lower in neonatal and young calves when stimulated with H-agg.IgG . These results indicate that CR3- and FcR-mediated phagocytic and killing activities of neutrophils in neonatal and young calves are different from those in cows . These phenomena may be associated with age-dependent changes in {Ca2+}i.

Prostaglandins Leukot Essent Fatty Acids, 1997 Apr, 56(4), 281 - 3
Effect of nordihydroguaretic acid and fluconazole on the LTC4/PGE2 ratio in the kidney of mice damaged by Candida albicans; Kustimur S et al.; The kidney is a major target organ in generalized candidiasis . When mice were infected with C.albicans, prostaglandin E2 (PGE2)-like activity was found to be significantly decreased while leukotriene C4 (LTC4)-like activity increased in the kidneys within 10 days . The aim of this study is to investigate the effect of nordihydroguaretic (NDGA) and fluconazole on the LTC4/PGE2 ratio in the mice kidneys infected by proteinase (+) C . albicans . The LTC4/PGE2 ratio was found to be significantly decreased both in NDGA and fluconazole-pretreated groups . These results indicate that pretreatment with the lipoxygenase inhibitor NDGA and the antifungal drug fluconazole, protect the kidney against C . albicans infection . These results also indicate a possible role of arachidonic acid metabolites (increase LTC4 and decrease PGE2) in kidney damage due to C . albicans infection.

J Med Microbiol, 1997 Apr, 46(4), 326 - 32
Scanning electron microscopy of the development of structured aerial mycelia and satellite colonies of phenotypically switched Candida albicans; Radford DR et al.; Candida albicans is an asexual diploid fungus that can express high frequency phenotypic switching . Switched variants can develop structured aerial mycelia (SAM) and cultures that have been grown for a protracted period exhibit satellite colonies . This study investigated the development of both SAM and satellite colonies by means of a freeze-drying technique and scanning electron microscopy . The results show that SAM may develop due to hyphae developing in the colony at the base of the structures . The development of satellite colonies is a result of cells that have grown deep into the agar re-emerging on to the agar/air surface and producing a new colony . Although both SAM and satellite colonies are often seen together on mature colonies, their cause and development are different.

Infect Immun, 1997 Apr, 65(4), 1335 - 44
Characterization of the Aspergillus nidulans aspnd1 gene demonstrates that the ASPND1 antigen, which it encodes, and several Aspergillus fumigatus immunodominant antigens belong to the same family; Calera JA et al.; For the first time, an immunodominant Aspergillus nidulans antigen (ASPND1) consistently reactive with serum samples from aspergilloma patients has been purified and characterized, and its coding gene (aspnd1) has been cloned and sequenced . ASPND1 is a glycoprotein with four N-glycosidically-bound sugar chains (around 2.1 kDa each) which are not necessary for reactivity with immune human sera . The polypeptide part is synthesized as a 277-amino-acid precursor of 30.6 kDa that after cleavage of a putative signal peptide of 16 amino acids, affords a mature protein of 261 amino acids with a molecular mass of 29 kDa and a pI of 4.24 (as deduced from the sequence) . The ASPND1 protein is 53.1% identical to the AspfII allergen from Aspergillus fumigatus and 48% identical to an unpublished Candida albicans antigen . All of the cysteine residues and most of the glycosylation sites are perfectly conserved in the three proteins, suggesting a similar but yet unknown function . Analysis of the primary structure of the ASPND1 coding gene (aspnd1) has allowed the establishment of a clear relationship between several previously reported A . fumigatus and A . nidulans immunodominant antigens.

Infect Immun, 1997 Apr, 65(4), 1321 - 6
A protective role of platelet-activating factor in murine candidiasis; Im SY et al.; Platelet-activating factor (PAF) is a potent phospholipid-derived modulator of immunological and inflammatory processes . In this study, the role of exogenous and endogenous PAF in resistance to infection with Candida albicans was investigated . Administration of PAF following a lethal challenge of C . albicans significantly protected mice from death and reduced the number of organisms in the kidneys . Neutralization of endogenous PAF with the PAF antagonist BN50739 shortened the mean survival time and increased the number of C . albicans cells per kidney . Shortly after infection of mice (30 min), significant levels of PAF were detected in the serum . PAF-induced protection appears to be mediated through the actions of tumor necrosis factor alpha (TNF-alpha), since pretreatment with anti-TNF-alpha before each injection of PAF abrogated the majority of PAF-induced enhanced resistance . Administration of PAF in vivo elevated serum TNF-alpha levels and TNF-alpha mRNA expression in the kidney . Production of TNF-alpha was markedly diminished by pretreatment with the PAF antagonist BN50739 prior to infection with C . albicans . We conclude that PAF, which is produced during infection with C . albicans, plays an important role in determining the level of resistance to this infectious microorganism . This effect of PAF appears to be mediated, at least in part, through the induction of TNF-alpha.

FEMS Microbiol Lett, 1997 Apr 1, 149(1), 59 - 64
Fabatins: new antimicrobial plant peptides; Zhang Y et al.; Two new antimicrobial peptides related to the gamma-thionine family have been isolated by acid extraction from the broad bean Vicia faba . The extract was separated by ion exchange chromatography, and a fraction showing antibacterial activity was further purified by reverse-phase HPLC . Material from a single HPLC peak was sequenced and revealed the presence of two peptides differing by one amino acid . The peptides were named fabatins . They are 47 amino acids long, have an overall positive charge and contain 8 cysteines that probably form 4 disulfide bridges characteristic of the gamma-thionins . Fabatins were active against both Gram-negative and Gram-positive bacteria, but were inactive against the yeasts Saccharomyces cerevisiae and Candida albicans.

FEMS Microbiol Lett, 1997 Apr 1, 149(1), 25 - 30
Differential inhibition of Candida albicans CYP51 with azole antifungal stereoisomers; Lamb DC et al.; Azole antifungal compounds are important in agriculture and in the treatment of mycotic infection . The target enzyme, sterol 14 alpha-demethylase (CYP51), is inhibited through binding of triazole N-4 to the haem of this P450, as a sixth ligand together with the N-1 substituent groups interacting in some way with the apoprotein . Here we use Saccharomyces cerevisiae expression systems for the target enzyme of Candida albicans to investigate binding of enantiomers of the azole antifungal compounds SCH39304 and tetraconazole . A molecular model produced previously provided qualitative explanations for these differences . Interaction of the azole antifungal aromatic group with Phe-233 or -235 may cause the higher activity for (R)-tetraconazole while inactivity of the (SS)-enantiomer of SCH39304 was predicted to result from incompatibility of the hydrophilic sulfonyl moiety when located into the hydrophobic pocket of the active site.

Appl Environ Microbiol, 1997 Apr, 63(4), 1312 - 7
Effect of nonylphenol on growth of Neurospora crassa and Candida albicans; Karley AJ et al.; The effects of the nonionic surfactant nonylphenol on the growth and morphologies of the filamentous fungus Neurospora crassa and the diploid yeast Candida albicans have been examined . Nonylphenol inhibited respiration and growth of N . crassa, effecting a 10-fold decrease in organism yield at 25 microM . Severe morphological defects were also induced: cell shape was abnormal and apical dominance was lost . Nonylphenol monoethoxylate (the parent compound of nonylphenol) was a less potent growth inhibitor and morphogen . The growth of the yeast form of C . albicans was sensitive to nonylphenol (inducing an order of magnitude decrease in specific growth rate with a 10-fold increase in dose concentration) but not nonylphenol monoethoxylate . Similarly, C . albicans ATP content was reduced and glucose-induced extracellular acidification was inhibited only by nonylphenol . Although estrogens may induce the dimorphic transition of C . albicans, nonylphenol (as an environmental estrogen mimic) failed to trigger germ tube formation under nonpermissive conditions and inhibited it under permissive conditions . The effects of nonylphenol are most readily explained as the result of uncoupling of respiration, which produces multiple physiological effects.

Clin Invest Med, 1997 Apr, 20(2), 85 - 93
Asymptomatic oral carriage of Candida albicans in patients with HIV infection; Fong IW et al.; OBJECTIVE: To assess the relationship of asymptomatic carriage of Candida albicans and clinically apparent thrush in patients with HIV infection . DESIGN: Prospective, longitudinal, controlled study . SETTING: The HIV clinic at St . Michael's Hospital, University of Toronto . PARTICIPANTS: One hundred and twenty-seven patients with HIV-infection were divided into 3 groups according to the CD4+ lymphocyte count, and 37 healthy volunteers served as controls . INTERVENTIONS: Determination of blood type, baseline CD4+ lymphocyte count in patients with HIV infection, and immunophenotyping . Samples of saliva (2 mL) were obtained from each patient and control . MAIN OUTCOME MEASURES: Carrier status, clinical presence of thrush, the association between carriage of C . albicans and blood type, secretor status and history of oral infection . RESULTS: In patients with HIV infection and C albicans colonization no correlation was found with blood type or secretor status of blood group antigen in the saliva . The frequency of oral carriage of yeast was greater in patients infected with HIV than in controls, but the difference was not significant for asymptomatic subjects with a CD4+ lymphocyte count greater than 500/microL . Persistent carriage of yeast and development of clinical thrush were associated with lower CD4+ counts . Clinical thrush developed only in patients with persistent asymptomatic carriage of C . albicans and CD4+ counts less than 500/microL . CONCLUSION: The greater risk of oral colonization with C . albicans in patients with HIV infection partly explains the high prevalence of thrush found in this group.

Antimicrob Agents Chemother, 1997 Apr, 41(4), 771 - 5
Improved activity of a synthetic indolicidin analog; Falla TJ et al.; A novel cationic peptide, CP-11, based on the structure of the bovine neutrophil peptide indolicidin, was designed to increase the number of positively charged residues, maintain the short length (13 amino acids), and enhance the amphipathicity relative to those of indolicidin . CP-11, and especially its carboxymethylated derivative, CP-11C, demonstrated improved activity against gram-negative bacteria and Candida albicans, while it maintained the activity of indolicidin against staphylococci and demonstrated a reduced ability to lyse erythrocytes . In Escherichia coli, CP-11 was better able than indolicidin to permeabilize both the outer membrane, as indicated by the enhancement of uptake of 1-N-phenylnaphthylamine, and the inner membrane, as determined by the unmasking of cytoplasmic beta-galactosidase, providing an explanation for its improved activity.

J Infect Dis, 1997 Apr, 175(4), 1012 - 5
Granulocyte colony-stimulating factor administered in vivo augments neutrophil-mediated activity against opportunistic fungal pathogens; Liles WC et al.; Granulocyte colony-stimulating factor (G-CSF) not only increases neutrophil (polymorphonuclear leukocyte, PMNL) production but also modulates PMNL biologic function . To assess the ability of G-CSF administered in vivo to enhance PMNL activity against opportunistic fungal pathogens, the antifungal activity of PMNL obtained from normal human volunteers before and after G-CSF administration was compared . In vivo, G-CSF significantly enhanced PMNL-mediated killing of Aspergillus fumigatus and Rhizopus arrhizus by 4-fold and 15-fold, respectively (P < .05) . In contrast, PMNL-mediated killing of Candida albicans was unaffected by G-CSF . The ability of aqueous fungal extracts to induce the PMNL respiratory burst was evaluated by luminol-enhanced chemiluminescence . G-CSF in vivo primed PMNL for sustained chemiluminescence in response to extracts of Candida, Aspergillus, and Rhizopus organisms . These data demonstrate that G-CSF in vivo augments antifungal activities of PMNL, thereby implicating a possible therapeutic role for G-CSF as a biologic response-modifying agent during opportunistic fungal infection.

J Bacteriol, 1997 Apr, 179(7), 2363 - 72
Isolation of the Candida albicans homologs of Saccharomyces cerevisiae KRE6 and SKN1: expression and physiological function; Mio T et al.; Cell wall beta-glucan in a pathogenic fungus, Candida albicans, is highly branched with beta-1,3 and beta-1,6 linkages . We have isolated the C . albicans cDNAs for KRE6 and SKN1, the genes required for beta-1,6-glucan synthesis in Saccharomyces cerevisiae . The results of Northern blot analysis revealed that C . albicans KRE6 was expressed at a higher level than SKN1 in the yeast phase, while SKN1 expression was strongly induced upon induction of hyphal formation . In addition, the C . albicans KRE6 and SKN1 mRNAs but not the actin mRNA were shortened during the yeast-hypha transition . Unlike S . cerevisiae, more than 50% of cell wall glucan was beta-1,6 linked in C . albicans . Neither beta-1,3-glucan nor beta-1,6-glucan was affected by the homozygous C . albicans skn1 delta null mutation . Although we never succeeded in generating the homozygous C . albicans kre6 delta null mutant, the hemizygous kre6 delta mutation decreased the KRE6 mRNA level by about 60% and also caused a more than 80% reduction of beta-1,6-glucan without affecting beta-1,3-glucan . The physiological function of KRE6 was further examined by studying gene regulation in C . albicans . When KRE6 transcription was suppressed by using the HEX1 promoter, C . albicans cells exhibited the partial defect in cell separation and increased susceptibility to Calcofluor White . These results demonstrate that KRE6 plays important roles in beta-1,6-glucan synthesis and budding in C . albicans.

J Bacteriol, 1997 Apr, 179(7), 2331 - 8
Overexpression of a cloned IMP dehydrogenase gene of Candida albicans confers resistance to the specific inhibitor mycophenolic acid; Kohler GA et al.; An IMP dehydrogenase gene was isolated from Candida albicans on a approximately 2.9-kb XbaI genomic DNA fragment . The putative Candida IMP dehydrogenase gene (IMH3) encodes a protein of 521 amino acids with extensive sequence similarity to the IMP dehydrogenases of Saccharomyces cerevisiae and various other organisms . Like the S . cerevisiae IMH3 sequence characterized in the genome sequencing project, the open reading frame of the C . albicans IMH3 gene is interrupted by a small intron (248 bp) with typical exon-intron boundaries and a consensus S . cerevisiae branchpoint sequence . IMP dehydrogenase mRNAs are detected in both the yeast and hyphal forms of C . albicans as judged by Northern hybridization . Growth of wild-type (sensitive) C . albicans cells is inhibited at 1 microg of mycophenolic acid (MPA), a specific inhibitor of IMP dehydrogenases, per ml, whereas transformants hosting a plasmid with the IMH3 gene are resistant to MPA levels of up to at least 40 microg/ml . The resistance of cells to MPA is gene dosage dependent and suggests that IMH3 can be used as a dominant selection marker in C . albicans.

MMWR Morb Mortal Wkly Rep, 1997 Mar 28, 46(12), 261 - 3
Candida albicans endocarditis associated with a contaminated aortic valve allograft--California, 1996; Oral lesions in Thai heterosexual AIDS patients: a preliminary study; Faculty of Dentistry, Prince of Songkla University, Haadyai, Songkhla, ThailandOBJECTIVE: To study the types of oral lesions in Thai heterosexual AIDS patients . DESIGN: Cross-sectional study and single centre . SETTING: Medical ward of the Prince of Songkla University Hospital, Thailand, from June 1994 to May 1995 . SUBJECTS: Heterosexual AIDS patients who had been admitted because of opportunistic infections and/or neoplasms . MAIN OUTCOME MEASURES: Types of oral lesions, opportunistic systemic diseases present and drugs in use, as well as the lymphocyte count, were recorded in each patient . Mycological investigations by the oral rinse technique were also performed . RESULTS: 41 patients were examined (32 male, 9 female; aged 19-53 years, median 29 years) . Oral lesions were found in 35 (85%) patients as follows: oral candidiasis (31), hairy leukoplakia (3), aphthous ulcer (3), linear gingival erythema (2), non-Hodgkin's lymphoma (2) and histoplasmosis (2) . CANDIDA ALBICANS: was isolated in 31 patients . There was correlation between the clinical signs of oral candidiasis and the colony-forming units of Candida (Mann-Whitney U test; two-tailed P = 0.0007) . CONCLUSIONS: Oral candidiasis was the most common lesion . It is of interest that non-Hodgkin's lymphoma was the only neoplasm detected . We conclude that oral lesions among Thai heterosexual AIDS patients may differ from those in other countries.

Gene, 1997 Mar 18, 187(2), 151 - 8
Isolation and characterization of a gene encoding alpha-tubulin from Candida albicans; Daly S et al.; A gene encoding the alpha-tubulin of Candida albicans has been cloned and characterized . Nucleotide sequence analysis reveals the presence of an intron within the structural gene and predicts the synthesis of a polypeptide of 448 amino acid residues . Comparison of nucleotide and amino acid sequences with the Saccharomyces cerevisiae alpha-tubulin encoding genes shows a 75% homology and about 92% similarity respectively . In contrast to S . cerevisiae, C . albicans appears to possess only one gene for alpha-tubulin which is able to functionally complement a S . cerevisiae cold-sensitive tub1 mutant.

Biochem Biophys Res Commun, 1997 Mar 17, 232(2), 350 - 3
Hemoglobin is utilized by Candida albicans in the hyphal form but not yeast form; Tanaka WT et al.; Hyphal cells of Candida albicans bound to human hemoglobin, but not the yeast cells . The amount of hemoglobin receptor was significantly higher on the hyphal cells than on the yeast cells . Only the hyphal cells of C . albicans used hemoglobin as a source of iron . The hemolytic factor was detected in the culture supernatant of the hyphal cell of C . albicans.

FEMS Microbiol Lett, 1997 Mar 15, 148(2), 247 - 54
Temperature-related expression of the vacuolar aspartic proteinase (APR1) gene and beta-N-acetylglucosaminidase (HEX1) gene during Candida albicans morphogenesis; Niimi M et al.; Expression of the Candida albicans vacuolar aspartic proteinase (APR1) and beta-N-acetylglucosaminidase (HEX1) genes was studied when carbon-starved cells of strains ATCC 10261 and A72 were induced to grow as yeast or as germ tube-forming cells . Amounts of APR1 mRNA were similar under yeast or germ tube growth conditions . However, more APR1 mRNA was present in cells grown at 28 degrees C than in cells grown at 37 degrees C . The Apr1 enzyme activity of cell-free extracts was not affected by cellular morphology, culture pH or growth temperature . Amounts of HEX1 mRNA were also higher in N-acetylglucosamine (GlcNAc)-induced cells grown at 28 degrees C than in cells grown at 37 degrees C . There was slightly more HEX1 mRNA in cells grown at pH 4.5 than in cells grown at pH 6.7 . The beta-N-acetylglucosaminidase activities of GlcNAc-grown cells correlated with the amounts of HEX1 mRNA and were higher when cells were grown at a lower temperature and at a lower pH . Although a similar temperature- and pH-dependent pattern of HEX1 mRNA expression was seen in cells grown on glucose, the enzyme activities in cell-free extracts were all very low . These data indicate that the APR1 and HEX1 genes play no direct role in the dimorphic transition of C . albicans and that transcription of both genes appears to be temperature regulated when the cells are released from carbon starvation . The expression of HEX1 mRNA is in part under the control of culture pH and translation of HEX1 mRNA seems to be regulated by glucose.

Nagoya J Med Sci, 1997 Mar, 60(1-2), 1 - 14
Strain-relatedness among different populations of the pathogenic yeast Candida albicans analyzed by DNA-based typing methods; Tanaka K; A pathogenic yeast Candida albicans is one of the most frequently isolated microorganisms in patients suffering from opportunistic infection . The typing of the isolates is important not only to elucidate the route of infection but to explore the evolution of the pathogenic yeast . This article presents an overview of recent research on the strain typing in different populations of C . albicans using the following DNA-based methods: electrophoretic karyotyping by pulsed field gel electrophoresis (PFGE), restriction fragment length polymorphism (RFLP) of digests of genomic DNA by restriction enzymes (with or without Southern hybridization with a DNA probe) and amplification of random or specific sequences by using polymerase chain reaction (PCR) . The results of these methods were compared for strain discrimination . Studies on the genetic diversity of the strains among different individuals in a single population cohort, and among different populations of healthy carriers and immunocompetent and immunocompromised patients, were reviewed . Typing of the strains recurrently isolated through the episodes of diseases such as vaginitis and AIDS revealed the occurrence of strain variation or microevolution . Typing of isolates from nosocomial infections indicated the possible occurrence of horizontal transmission of the disease within a single hospital . In addition, recent studies suggested a possible mechanism that might involve the C . albicans-specific repetitive sequences, RPSs, for chromosome rearrangements leading to strain variation.

Hautarzt, 1997 Mar, 48(3), 175 - 80
{Diagnosis of dermatomycoses with polymerase chain reaction}; Bock M et al.; A polymerase chain reaction (PCR) assay for the detection of dermatophytes was established . The primers "TR1" and "TR2" amplify a 581 bp fragment within the gene coding for the small ribosomal subunit (185 rRNA) of fungi . PCR allowed the detection of isolates of 7 common dermatophytes and in addition several yeasts and moulds . Hybridisation with specific oligonucleotides results in the identification of dermatophytes and Candida albicans . Restriction analysis of the PCR product allowed us to distinguish between dermatophytes and yeasts or moulds . The specificity of the PCR with respect to fungi was assessed by testing human DNA collected from 42 dermis and epidermis specimens as well as DNA from selected plants and animals . To evaluate the clinical relevance of the PCR assay, 69 routinely collected skin and nail specimens were examined by PCR and culture . PCR detected dermatophytes in 35 and culture in 28 of 38 specimens that were classified as positive . Sensitivity of PCR (92%) was higher than that of culture (73%) . These results show that PCR has advantages over culture for the detection of dermatophytes.

J Med Vet Mycol, 1997 Mar-Apr, 35(2), 133 - 7
New evidence that Candida albicans possesses additional ATP-binding cassette MDR-like genes: implications for antifungal azole resistance; Walsh TJ et al.; Emergence of resistance of Candida albicans to antifungal triazoles is increasingly recognized as an important cause of refractory mucosal candidiasis in HIV-infected patients . Recently, CDR1, which is thought to be analogous to the human MDR-1 P-glycoprotein, has been cloned in C . albicans . It has been proposed that its expression is partially responsible for fluconazole resistance in C . albicans . This gene is characterized by the presence of an ATP binding cassette (ABC) region and is distinct from the BENr gene which does not encode such a functional domain . As the molecular basis for fluconazole resistance appears to be multifactorial, we considered that there may be other ATP binding cassette-containing MDR genes that may potentially contribute to antifungal azole resistance in C . albicans . We therefore sought to identify potential target sequences that may be derived from candidate genes that share homology with the ATP binding cassette region of the human MDR-1 P-glycoprotein . Degenerate oligonucleotide primers based on the known sequence from the ATP binding cassette region of the human MDR-1 P-glycoprotein were used to amplify PCR products within the range of 100 bp in length from C . albicans isolates (3 fluconazole-susceptible and 3 fluconazole-resistant) . Sequence analysis of individually subcloned PCR products, derived from the six isolates revealed 34 sequences in total . The results of our study identified 14 clones (with at least one per isolate) with a high degree of homology to the ATP binding cassette of the human MDR-1 P-glycoprotein . The BLAST search did not disclose homology of these new sequences to the C . albicans CDR1 gene, suggesting that C . albicans may possess more than one MDR-like gene . We conclude that C . albicans may possess one or more additional genes encoding ATP binding cassette MDR-like proteins that are distinct from CDR 1 and which could participate in the development of fluconazole resistance.

J Med Vet Mycol, 1997 Mar-Apr, 35(2), 115 - 21
Atypical strains of Candida albicans recovered from AIDS patients; Pujol C et al.; By using multilocus enzyme electrophoresis (MLEE) we have analyzed the genetic diversity encountered among chlamydospore-positive Candida albicans strains . While the type II strains of the former C . stellatoidea were genetically indistinguishable from those of C . albicans, type I strains constituted a distinct subgroup compared with C . albicans strains . Nevertheless, all these strains remained genetically very closely related compared with other species of Candida (e.g . C . tropicalis, C . krusei and C . glabrata) . These results corroborate the synonymy between C . stellatoidea and C . albicans . Chlamydospore-positive C . albicans strains with atypical sugar assimilation patterns displayed a great genetic divergence from the cluster constituted by C . albicans and the strains of the former C . stellatoidea . However, these atypical strains were more closely related to C . albicans than they were to C . tropicalis, C . krusei or C . glabrata . These strains represent a genetically entity distinct from the typical C . albicans strains used in this study . The data also support the view that the atypical strains described here belong to the same genetic group as atypical C . albicans strains previously described by others.

J Med Vet Mycol, 1997 Mar-Apr, 35(2), 87 - 99
Molecular bases of adhesion of Candida albicans; Fukazawa Y et al.; The purpose of this review is to focus on the location and the adhesion activity of the protein (peptide) and the mannan moieties of the mannoprotein in the outer surface of the Candida albicans cell wall . A macromolecule of the mannoprotein located on the outermost surface is undoubtedly a strong adhesin comprising several adhesion molecules including protein and mannan . Mannoproteins can be divided into two classes, higher molecular weight peptidomannans (260 kDa) and lower molecular weight mannoproteins (50-66 kDa), both of which consist of similar mannans and disparate proteins or peptides which have distinct adhesion specificities . The protein moiety of mannoprotein can be divided functionally into two groups, lectin-like proteins and proteins recognizing arginine-glycine-aspartic acid (RGD) ligands . The latter proteins are further subdivided into two groups, CR2/CR3-like proteins and proteins binding extracellular matrix (ECM) proteins . Hydrophobicity of the cell surface of C . albicans influences adhesion of the organisms to epithelial cells . Degree of glycosylation of cell surface mannoproteins that affect yeast cell surface hydrophobicity affects adhesion of C . albicans to epithelial cells . The hydrophobic proteins may have low levels of glycosylation, and changes in glycosylation may determine exposure of hydrophobic protein regions at the cell surface . The serotype A-specific oligosaccharide of antigen 6 (pentaose or hexaose of mannan moiety) has been shown to exhibit marked adhesion ability for epithelial cells, and mannotetraose related to antigenic factor 5 which is present in both serotypes A and B showed adhesive activity for tissue macrophages . Proteinoceous adhesins of C . albicans are expressed preferably on the mycelial form . It is suggested that several of the adhesion molecules of C . albicans described above appear to complementarily utilize multiple adhesion mechanisms.

J Med Vet Mycol, 1997 Mar-Apr, 35(2), 79 - 86
Lipopeptide inhibitors of fungal glucan synthase; Kurtz MB et al.; The echinocandins and pneumocandins are lipopeptide antifungal agents that inhibit the synthesis of 1,3-beta-D-glucan, an essential cell wall homopolysaccharide found in many pathogenic fungi . Compounds with this fungal-specific target have several attractive features: lack of mechanism-based toxicity, potential for fungicidal activity and activity against strains with intrinsic or acquired resistance mechanisms for existing antimycotics . Semi-synthetic analogues of naturally occurring lipopeptides are currently in clinical trials with the aim of treating systemic candidiasis and aspergillosis . Thus a fuller understanding of the target enzyme and its inhibition by these compounds should be useful for epidemiological and other clinical studies . Although it has been long known that lipopeptides inhibit fungal glucan synthase activity both in cell extracts and in whole cells, the genetic and biochemical identification of the proteins involved has been accomplished only recently . We now know that in Saccharomyces cerevisiae, glucan synthase is a heteromeric enzyme complex comprising one large integral membrane protein (specified by either FKS1 or by FKS2) and one small subunit more loosely associated with the membrane (specified by RHO1) . Additional components may also be involved . The heteromeric enzyme complex containing Fks1p constitutes the majority of the activity found in vegetatively growing cells in this organism . The FKS2 gene product is needed for sporulation . Lipopeptides affect the function of the Fksp component from either FKS gene . The current model for interaction and regulation of these components in S . cerevisiae and the application to Candida albicans and other pathogenic fungi are discussed in this review.

Clin Lab Haematol, 1997 Mar, 19(1), 39 - 47
Effects of in vitro and in vivo cytokine treatment, leucapheresis and irradiation on the function of human neutrophils: implications for white blood cell transfusion therapy; Cohen DM et al.; Treatment of human polymorphonuclear leucocytes (PMNL) separated by density sedimentation (DS) from normal donors (PMNL-NL-DS) with interferon-gamma (IFN-gamma) + granulocyte colony-stimulating factor (G-CSF) lessens the damage caused by isolation and irradiation . We have studied granulocyte-macrophage colony-stimulating factor (GM-CSF) in this system, as well as the behaviour of PMNL collected by continuous flow leucapheresis (CFL) from donors treated with G-CSF (PMNL-GCSF-CFL) . After isolation, PMNLs were treated with IFN-gamma + G-CSF, GM-CSF or IFN-gamma + G-CSF + GM-CSF, irradiated with 0 or 30 Gy and studied after 0 and 20 h in cell culture . All regimens reduced apoptosis of PMNL-NL-DS . Killing of Candida albicans by 20-h-old PMNL-NL-DS was best preserved by IFN-gamma + G-CSF treatment . A similar pattern of results was obtained for assays of PMNL-NL-DS chemotaxis and superoxide production . There was a consistent trend toward reduced function after irradiation in all assays . PMNL-GCSF-CFL less often demonstrated the morphological features of apoptosis, and this was further reduced by cytokine regimens containing IFN-gamma + G-CSF . In assays of C . albicans killing and chemotaxis, 20-h-old untreated PMNL-GCSF-CFL performed as well as freshly isolated PMNL-GCSF-CFL . PMNL-GCSF-CFL showed decay in CD11b (CR3), CD16 (Fc gamma III) and CD64 (Fc gamma R1) expression after 20 h in cell culture, but treatment with IFN-gamma + G-CSF preserved expression . There was a trend toward reduced function after radiation . Comparison of PMNL-GCSF separated by CFL and DS demonstrated that CFL itself is a strong inducer of the morphological features of apoptosis . This study shows that while separation by CFL, and irradiation are damaging to PMNLs, damage may be reduced by use of cytokines.

Eur J Clin Microbiol Infect Dis, 1997 Mar, 16(3), 228 - 32
Evaluation of the E test for fluconazole susceptibility testing of Candida albicans isolates from oropharyngeal candidiasis; Dannaoui E et al.; The aim of the present study was to evaluate the utility of the E test in determining the antifungal susceptibility of Candida albicans . Reproducibility of the E test was determined for amphotericin B, fluconazole, and itraconazole using three different solid media: RPMI 1640, Casitone, and yeast nitrogen base agar . Minimum inhibitory concentrations (MICs) were comparable (results at +/- 2 dilutions) in 92% of the tests for amphotericin B and in 100% for fluconazole and itraconazole . Determination of MIC endpoints was easiest on Casitone agar . Candida albicans isolates from 23 patients undergoing fluconazole therapy for oropharyngeal candidiasis were tested for fluconazole susceptibility . Good correlation was obtained between the MICs of fluconazole and clinical outcome . Clinical failure was associated with strains for which MICs were > or = 48 micrograms/ml . These results suggest that the E test has potential utility for fluconazole susceptibility testing of clinical yeast isolates.

Eur J Biochem, 1997 Mar 1, 244(2), 325 - 33
Identification of an Fe(III)-dihydroxyphenylalanine site in recombinant phosphomannose isomerase from Candida albicans; Smith JJ et al.; Candida albicans phosphomannose isomerase (PMI) is a zinc metalloprotein of known crystal structure . When heterologously overexpressed in Escherichia coli, a blue protein that contains up to 0.5 iron atoms/PMI molecule could be isolated, with absorption maxima at 420 nm and 680 nm . These bands are reminiscent of ferric catecholate complexes, an assignment that has been confirmed by resonance Raman spectroscopy, and by reaction with Arnow's reagent, which is specific for the presence of 3,4-dihydroxyphenylalanine (Dopa) . After enzymatic digestion of blue PMI, a peptide with the sequence DPHAXISG was isolated corresponding to residues Asp283-Gly290 in the amino acid sequence of C . albicans PMI, where the unidentified residue X287 is encoded by a tyrosine codon . It is proposed that iron and oxygen bring about hydroxylation of Tyr287 in PMI and that Fe(III) subsequently chelates the Dopa residue to give the characteristic absorption spectrum . The EPR spectrum of the blue protein suggests three iron environments in the protein, two in axial environments with E/D values approximately equal to 0.06 and 0.12 and one rhombic species . The nature of the iron co-ordination sites is discussed with the help of model systems and by comparison with other blue non-heme iron proteins.

Br J Dermatol, 1997 Mar, 136(3), 319 - 25
Differential T-cell reactivity to the round and oval forms of Pityrosporum in the skin of patients with psoriasis; Baker BS et al.; Pityrosporum yeasts have been implicated as a trigger for the initiation of scalp lesions in psoriasis . To determine whether Pityrosporum-reactive T cells are present in lesional psoriatic skin . T-cell lines (TCL) were cultured from the scalps of nine patients with psoriasis and seven with alopecia areata (disease controls), and from non-scalp lesions from six of the psoriatic patients . The psoriatic skin TCL were stained for CD3, CD4, CD8 and TCR alpha beta expression and tested in a proliferation assay with Candida albicans and purified protein derivative (PPD), and cytoplasmic and cell-wall extracts of P . ovale (oval) and P . orbiculare (round) . The proliferative responses of corresponding peripheral blood mononuclear cells (PBMC) were also determined . All the PBMC samples responded to the Pityrosporum extracts to variable extents, but no significant difference in the response of the group to the two different forms of yeast was observed . The response was mediated by CD4+ T cells and inhibited by the addition of anti-HLA-DR antibody . In addition, all nine psoriatic scalp TCL, which were predominantly CD3+, CD4+ TCR alpha beta(+), responded to the cytoplasmic, and five of nine TCL to the cell-wall extract of P . orbiculare . In contrast, only three of the nine TCL proliferated to either extract of P . ovale . This difference was significant for both the cytoplasmic (P < 0.01) and cell wall (P = 0.01) extracts . Similarly, the TCL cultured from non-scalp psoriatic lesions also showed a more marked response to the P . orbiculare extracts (P = 0.05) . Furthermore, four of seven and two of seven scalp TCL from lesions of alopecia areata responded to the P . orbiculare and P . ovale extracts, respectively; these responses did not differ significantly from those of the psoriatic scalp TCL . None of the skin TCL responded to either Candida albicans or PPD . These findings demonstrate that T cells with differential reactivity to the round and oval forms of Pityrosporum are present in, but are not specific for, psoriatic skin lesions . A role for these cells in the pathogenesis of psoriasis remains speculative.

Infection, 1997 Mar-Apr, 25(2), 112 - 6
Multifocal osteoarthritis due to Candida albicans in a neonate: serum level monitoring of liposomal amphotericin B and literature review; Evdoridou J et al.; A 32-week neonate weighing 2,300 g at birth with fungemia due to Candida albicans subsequently developed multifocal osteoarthritis of the lower extremities due to the same organism during therapy with amphotericin B (0.5 mg/kg/day) and flucytosine (100 mg/kg/day) to which the isolates were susceptible . Liposomal amphotericin B (3.5 mg/kg/day) was substituted for conventional amphotericin B and complete clinical and radiologic recovery as well as sterilization of affected joints were achieved with a 38-day-course (144.5 mg/kg total) . Adequate drug concentrations in serum and synovial fluid were attained with liposomal amphotericin B . Nine neonates (< or = 28 days of age) with bone and/or joint infections due to Candida spp . had previously been reported in the English-language literature . To our knowledge, the present case is the first reported cure with liposomal amphotericin B . Previous cases are reviewed and the potential role of liposomal amphotericin B in this serious neonatal infection is discussed.

Medicine (Baltimore), 1997 Mar, 76(2), 94 - 103
Fungal prosthetic valve endocarditis in 16 patients . An 11-year experience in a tertiary care hospital; Melgar GR et al.; Fungal prosthetic valve endocarditis (PVE) is an infrequent but serious complication of valve replacement surgery . To examine its long-term outcome, we retrospectively studied 16 patients with 19 episodes of definite fungal PVE . The mean age was 51 years (range, 27-71 yr) . Onset of fungal PVE ranged from 8 days to 3.4 years after valve replacement . Candida albicans was the most common (56%) pathogen isolated . A portal of entry was identified in only 25% of the patients; the presence of intravascular catheters (50%) and prior bacterial endocarditis (38%) were leading predisposing factors . Fever (83%) was the most consistent clinical finding . Potentially serious embolic events, particularly strokes (32%), were common at presentation . Transesophageal echocardiography (sensitivity = 100%) was more useful than transthoracic echocardiography (sensitivity = 60%) in detecting lesions due to fungal PVE . Combined valve replacement surgery and amphotericin B (mean total dose of 1.8 g) in 15 patients resulted in an 87% in-hospital survival and 67% overall survival with a mean follow-up of 4.5 years (range, 5 mo to 16 yr) . Two patients had 3 late relapses of fungal PVE up to 9 years after the preceding episode . Each relapse was treated with repeat valve replacement and amphotericin B; in addition, oral azole was utilized for chronic suppression, although the efficacy of this strategy remains unproven . Because of the possibility of relapse, long-term follow-up is essential even after surgical and prolonged antifungal therapy.

Ann Clin Lab Sci, 1997 Mar-Apr, 27(2), 151 - 6
The effect of estradiol on Candida albicans growth; Gujjar PR et al.; The size of colonies growing on Sabouraud's dextrose agar (SDA) supplemented with 1 microM beta estradiol (1,3,5{10}-estratiene-3,17B-diol) was compared to those growing without the estrogen supplement . Data (video pixel counts) were obtained by video capture and image analysis and revealed that yeast strains were not uniformly stimulated by estradiol . One estrogen-responsive strain was evaluated in a chemically defined medium to evaluate growth in the presence of 17-alpha and 17-beta isomers of estradiol . The beta isomer promoted more rapid growth of the test organism and resulted in greater biomass production than the alpha isomer . Understanding of the role of mammalian hormones as growth promoting signals may provide information on this organism's shift from saprophytic to commensal to pathogenic status in vivo . Strain variation in response to estrogen may provide an explanation for differences in virulence.

Microbiology, 1997 Mar, 143 ( Pt 3), 733 - 8
High resolution X-ray micrography of live Candida albicans using laser plasma pulsed point X-ray sources; Rajyaguru JM et al.; Electron microscopy is still the most frequently used method for visualization of subcellular structures in spite of limitations due to the preparation required to visualize the specimen, High resolution X-ray microscopy is a relatively new technique, still under development and restricted to a few large synchrotron X-ray sources . We utilized a single-shot laser (nanosecond) plasma to generate X-rays similar to synchrotron facilities to image live cells of Candida albicans . The emission spectrum was tuned for optimal absorption by carbon-rich material . The photoresist was then scanned by an atomic force microscope to give a differential X-ray absorption pattern . Using this technique, with a sample image time of 90 min, we have visualized a distinct 152.24 nm thick consistent ring structure around cells of C albicans representing the cell wall, and distinct 'craters' inside, one of 570-90 nm diameter and three smaller ones, each 400 nm in diameter . This technique deserves further exploration concerning its application in the ultrastructural study of live, hydrated microbiological samples and of macromolecules.

Microb Pathog, 1997 Mar, 22(3), 187 - 92
Evidence that two independent host genes influence the severity of tissue damage and susceptibility to acute pyelonephritis in murine systemic candidiasis; Ashman RB et al.; Tissue damage in the kidney and brain after systemic infection with Candida albicans was examined in recombinant inbred strains (AKXL) derived from AKR and C57/L progenitors . Nine of the 15 strains showed mild (C57/L-like) tissue damage . Of the remainder, two strains developed lesions comparable to the AKR parental strain, whereas four exhibited a much more severe pattern of tissue damage . This was characterized by pronounced mycelial growth in the brain, and gross oedema of the kidney, with extensive fungal colonization and marked tissue destruction . The presence of the null allele of the haemolytic complement gene (Hc) may be necessary, but not sufficient, for the expression of the very severe lesions . The results were interpreted as reflecting the actions of two independent genes, which have been designated Carg1 and Carg2 (Candida albicans resistance genes 1 and 2).

J Prosthet Dent, 1997 Mar, 77(3), 306 - 12
Adherence of Candida albicans to experimental denture soft lining materials; Waters MG et al.; STATEMENT OF PROBLEM: Colonization of denture soft lining materials by Candida albicans can result in clinical problems . Two experimental silicone soft lining materials have been developed and demonstrate good physical properties . PURPOSE OF STUDY: The aim of this study was to determine the extent of candidal adherence to these materials compared with a commercially available soft lining material and an acrylic resin denture base . MATERIAL AND METHODS: The experimental materials were constructed in a stainless-steel mold, and their surface energies were determined with a dynamic contact angle analyzer . The adherence of three strains of C . albicans was determined with an in vitro assay . In addition, one test strain was used to determine the effect on adherence of precoating the materials with whole saliva . RESULTS: Adhesion to all materials was strain variable, with the lowest adherence recorded for the two experimental materials . Decreased adherence was also apparent after precoating the materials with saliva . Correlation between surface energy of the material and the degree of candidal adhesion was seen for one strain, but no correlation was seen for the other two strains . CONCLUSION: Adherence of C . albicans to the two experimental silicone soft lining materials was significantly less than that for an acrylic resin denture base and a commercially available soft lining material.

Clin Exp Immunol, 1997 Mar, 107(3), 451 - 7
Responsiveness of human polymorphonuclear cells (PMNL) to stimulation by a mannoprotein fraction (MP-F2) of Candida albicans; enhanced production of IL-6 and tumour necrosis factor-alpha (TNF-alpha) by MP-F2-stimulated PMNL from HIV-infected subjects; Torosantucci A et al.; IL-1beta, IL-6, IL-8 and TNF-alpha production by PMNL from 21 HIV-infected (HIV+), including 11 full-blown AIDS, and 20 HIV-uninfected (HIV-) subjects (matched for age and sex to HIV+ ones) was studied by reverse transcriptase-polymerase chain reaction (RT-PCR) and ELISA . PMNL from both categories of subjects were strongly stimulated in their actual cytokine production by a mannoprotein fraction (MP-F2) of Candida albicans, as well as by the bacterial lipopolysaccharide (LPS) . These stimulatory effects were apparently due to increased cytokine gene expression and were substantially reversed by the physiological inhibitor IL-10 . However, PMNL from HIV+ subjects showed increased IL-6 and TNF-alpha gene expression and produced more IL-6 and TNF-alpha than PMNL from HIV- controls, under similar stimulation conditions . This difference could not be attributed to a given stage of HIV infection, any associated medication, or to a generalized increase of gene expression, as quantitatively similar beta-actin and IL-1beta transcripts were detected . Moreover, no significant difference in IL-8 production by the PMNL from HIV+ and HIV- subjects was observed . Our studies suggest that PMNL from HIV+ subjects might add to other cellular sources of IL-6 and TNF-alpha (e.g . monocytes-macrophages) in contributing to the cytokine-dysregulated pattern typical of the HIV+ patient.

Int Arch Allergy Immunol, 1997 Mar, 112(3), 313 - 6
IgE antibody response against Aspergillus umbrosus in farmer's lung disease; Kaukonen K et al.; IgG and IgA antibodies against fungus Aspergillus umbrosus have been found in the sera of patients with farmer's lung (FL) disease and healthy exposed farmers in Finland . To determine the IgE response to antigens of A . umbrosus and Candida albicans, sera from 20 patients with FL, 20 healthy farmers and 20 nonfarming controls were tested by nitrocellulose radioallergosorbent test (RAST) . The values of RAST indices were low in each group and the only statistically significant difference was found between the groups of FL patients and nonfarming controls against A . umbrosus polysaccharide antigen . Individual IgE responses to polysaccharide antigens of A . umbrosus and C . albicans correlated among FL patients, however, no cross-reacting IgE antibodies could be shown . To conclude, the IgE antibody levels of A . umbrosus polysaccharide and crude antigens were low in FL patients, healthy exposed farmers and nonfarming controls . Neither polysaccharide nor crude antigen-specific IgE antibodies of A . umbrosus have any diagnostic value in FL disease.

Antimicrob Agents Chemother, 1997 Mar, 41(3), 575 - 7
In vitro activities of voriconazole (UK-109,496) against fluconazole-susceptible and -resistant Candida albicans isolates from oral cavities of patients with human immunodeficiency virus infection; Ruhnke M et al.; The susceptibility of Candida albicans to a new antifungal triazole, voriconazole (UK-109,496), was investigated in 105 isolates obtained from the oral cavities of patients with human immunodeficiency virus (HIV) infection to study this drug's activity against fluconazole-susceptible and -resistant isolates . MICs were determined by a broth microdilution technique according to document M27-T from the National Committee for Clinical Laboratory Standards and by using a broth microdilution technique and a synthetic high-resolution medium . These antifungal susceptibility testing methods showed high levels of agreement (93% for fluconazole and 86% for voriconazole) . Data from in vitro studies showed that voriconazole has good activity against fluconazole-susceptible and -resistant C . albicans isolates; the MICs at which 90% of all isolates were inhibited were 0.19 to 0.39 microgram/ml . We found that for isolates for which fluconazole MICs were high, voriconazole MICs were proportionally higher than those for fluconazole-susceptible C.albicans (P < 0.001) . Pretreatment isolates from six patients with fluconazole-refractory esophageal candidiasis were included in the study . For these isolates the MICs were < or = 0.39 microgram/ml, and all patients responded to voriconazole . These results suggest that voriconazole is effective even in the treatment of fluconazole-refractory esophageal candidiasis and should be studied further to determine its clinical relevance in patients with HIV infection.

Antimicrob Agents Chemother, 1997 Mar, 41(3), 535 - 9
Reversible fluconazole resistance in Candida albicans: a potential in vitro model; Calvet HM et al.; To study the development and potential mechanisms of antifungal resistance in relation to antifungal exposure, reversible fluconazole resistance was examined in vitro . Candida albicans ATCC 36082 blastospores were passed in liquid yeast nitrogen base medium containing either 4, 8, 16, or 128 micrograms of fluconazole per ml, and susceptibility testing was performed after each passage . High-level fluconazole resistance (50% inhibitory concentration, > 256 micrograms/ml) developed in the isolates after serial passage in medium containing 8, 16, or 128 micrograms of fluconazole per ml, but not in isolates passed in 4 micrograms of fluconazole per ml . Reduced susceptibility was noted within four to seven passages, which was equivalent to 14 to 19 days of exposure to the drug . However, all isolates returned to the susceptible phenotype after 8 to 15 passages in medium lacking the drug; thus, fluconazole resistance was reversible in vitro . In vivo, organisms retained the resistant phenotype after a single passage in the rabbit model of infective endocarditis . Restriction digest profiles and karyotypic analysis of the parent strain and selected fluconazole-resistant and -susceptible isolates from each group were identical . Investigations into the molecular mechanisms of this reversible resistance failed to reveal increased accumulation of mRNA for 14 alpha-demethylase, the target enzyme for fluconazole, or for the candidal multidrug transporters CDR1 and BENr . This process of continuous in vitro exposure to antifungal drug may be useful as a model for studying the effects of different antifungal agents and dosing regimens on the development of resistance and for defining the mechanism(s) of reversible resistance.

J Clin Microbiol, 1997 Mar, 35(3), 667 - 72
Identification of various medically important Candida species in clinical specimens by PCR-restriction enzyme analysis; Morace G et al.; A single primer pair amplifying a cytochrome P-450 lanosterol-14 alpha-demethylase (L1A1) gene fragment that encodes a highly conserved region was used to detect yeast DNA in clinical specimens . Positive PCR products were obtained from genomic DNAs of Candida albicans, C . parapsilosis, C . tropicalis, C . guilliermondii, C . krusei, C . (Torulopsis) glabrata, and C . kefyr . No human, bacterial, or parasitic DNA was amplified . The sensitivity was evaluated for C . albicans genomic DNA by using various DNA concentrations (200 pg to 2 fg) . The amplified DNAs of Candida species with unknown P-450 L1A1 gene sequences were subcloned and sequenced . Identification at the species level was achieved by digestion of the PCR products with different restriction enzymes . A specific restriction enzyme analysis pattern was determined for each species investigated . Subsequently, we used PCR to detect specific yeast DNA directly with clinical specimens such as blood and bronchoalveolar lavage specimens . After appropriate treatment, the specimens were processed by PCR and the results were compared with those obtained by traditional diagnostic procedures such as cultures and serology . Although preliminary, the PCR results seem to correlate well, at least for blood, with those of antigen detection assays and traditional blood cultures, with a better and earlier detection of candidemia.

Infect Immun, 1997 Mar, 65(3), 1077 - 82
Involvement of mannose receptor in cytokine interleukin-1beta (IL-1beta), IL-6, and granulocyte-macrophage colony-stimulating factor responses, but not in chemokine macrophage inflammatory protein 1beta (MIP-1beta), MIP-2, and KC responses, caused by attachment of Candida albicans to macrophages; Yamamoto Y et al.; The production of chemotactic cytokines (chemokines) and other cytokines by macrophages in response to fungal infection is thought to be critical during the course of candidiasis . However, the mechanism of cytokine synthesis by macrophages in response to fungi is not well understood . Therefore, the response of macrophages to Candida albicans was examined in terms of receptor-mediated chemokine and other cytokine mRNA induction . Attachment of C . albicans to murine thioglycollate-elicited peritoneal macrophages induced increased mRNA levels of the cytokines interleukin-1beta (IL-1beta), IL-6, and granulocyte-macrophage colony-stimulating factor (GM-CSF) and the chemokines macrophage inflammatory protein 1beta (MIP-1beta), MIP-2, and KC (a member of the platelet factor 4 neutrophil chemoattractant family), as determined by quantitative reverse transcription-PCR . However, treatment of macrophages with alpha-methyl-D-mannoside significantly reduced the cytokine GM-CSF response to C . albicans but did not affect the chemokine MIP-2 response . Antisense oligodeoxynucleotide (ODN) to mannose receptor (MR) mRNA inhibited the expression and function of MR in macrophages as determined by Western blot analysis and 125I-labeled mannose-bovine serum albumin (BSA) binding, and also inhibited the elevation of cytokine IL-1beta, IL-6, and GM-CSF mRNA levels induced by C . albicans attachment . Elevation of chemokine MIP-1beta, MIP-2, and KC mRNA levels induced by C . albicans was not affected in macrophages whose MR expression was suppressed by antisense ODN treatment . Furthermore, IL-4 treatment of macrophages, which up-regulated MR expression as determined by Western blot analysis and fluorescein isothiocyanate-labeled mannose-BSA uptake, enhanced the level of cytokine GM-CSF mRNA induced by C . albicans but not the level of the chemokine MIP-2 mRNA . These results indicate that selected cytokine responses of macrophages to C . albicans are mediated by MR, while some chemokine responses may be mediated by other receptors.

Infect Immun, 1997 Mar, 65(3), 1023 - 31
Purification and in vitro activities of rabbit platelet microbicidal proteins; Yeaman MR et al.; Recent in vitro studies have demonstrated that rabbit platelets release a small, cationic antimicrobial protein in response to thrombin stimulation under physiological conditions (M . R . Yeaman, S . M . Puentes, D . C . Norman, and A . S . Bayer, Infect . Immun . 60:1202-1209, 1992) . This observation prompted our present investigation, focused on determining the array of antimicrobial proteins contained within rabbit platelets and their in vitro activity against common bloodstream pathogens . A group of small (6.0- to 9.0-kDa), cationic proteins with in vitro antimicrobial activity was purified from whole and thrombin-stimulated rabbit platelets by gel filtration and reversed-phase high-performance liquid chromatography . Purified proteins in micromolar concentrations (10 to 40 microg/ml) exerted in vitro microbiostatic and/or microbicidal activities against Staphylococcus aureus, Escherichia coli, and Candida albicans in a dose-dependent manner . The antimicrobial activities of proteins purified from rabbit platelet acid extracts were generally inversely related to pH, with maximal activity observed at pH 5.5 . In contrast, the predominant protein isolated from thrombin-stimulated rabbit platelets, though biochemically and microbiologically similar to proteins extracted by acid, exhibited antimicrobial activities which were modestly enhanced at pH 7.2 compared with pH 5.5 . Amino acid compositional analyses in combination with molecular mass determinations suggest that the majority of these proteins are distinct molecules not derived from a single common precursor . Collectively, these data indicate that rabbit platelets contain proteins which exert potent in vitro antimicrobial activity against bacterial and fungal pathogens which commonly invade the bloodstream . Moreover, several of these proteins were released from platelets stimulated with thrombin under physiological conditions and exerted potent antimicrobial activities in physiological pH ranges . These observations support the hypothesis that platelets serve an important role in host defense against infection, via localized release of antimicrobial proteins in response to stimuli associated with tissue injury or microbial colonization.

J Immunol, 1997 Mar 1, 158(5), 2356 - 62
An immunoregulatory role for neutrophils in CD4+ T helper subset selection in mice with candidiasis; Romani L et al.; Granulocytes may serve immunoregulatory and effector roles in different limbs of the immune response to infection . Using live vaccine strain or virulent challenge in mucosal or systemic infection of mice with Candida albicans, we examined the effect of mAb-mediated depletion of neutrophils on the course of primary and secondary challenge and on development of CD4+ cell-dependent immunity . We obtained evidence of deleterious effects of neutrophil depletion occurring at the time of infection under all conditions of testing, both in naive and in previously immunized mice . In contrast, neutrophil depletion appeared to benefit the hosts late in the course of an overwhelming systemic infection . In an attempt to correlate neutrophil function with the nature of the T cell response, we tested the ability of neutrophils to produce cytokines associated with functionally distinct CD4+ Th cell responses to Candida . We found that neutrophils were endowed with the capacity to secrete IL-12 and IL-10 in vitro in response to the yeast . Neutrophil ablation early in the course of Th1-associated, self-limiting infection appeared to change the qualitative development of the T cell response, and rendered mice susceptible to infection . In addition to long recognized contributions to acute anti-candidal responses, these data suggest an important role for neutrophils both in initiation and in expression of Candida-specific immunity.

J Immunol, 1997 Mar 1, 158(5), 2294 - 302
Early T cell unresponsiveness in mice with candidiasis and reversal by IL-2: effect on T helper cell development; Spaccapelo R et al.; To investigate the role and effect of IL-2 in the genesis of Th1 and Th2 responses to Candida albicans in vivo, we assessed the levels of IL-2 production and the Ag-specific proliferative response in mice with healing or nonhealing infection and the effects of IL-2 neutralization or administration on the course and outcome of infection and on the type of CD4+ Th immunity elicited . High levels of IL-2 production and Ag-specific proliferation in vitro correlated with disease progression in susceptible mice . In contrast, resolution of infection in resistant mice was accompanied by the induction of Ag-specific hyporesponsiveness and impaired IL-2 production . Progression of infection did not occur in susceptible mice treated with anti-IL-2 or anti-IL-2R mAbs; conversely, disease resolution was prevented in resistant mice treated with IL-2 . CD4+ Th1 cell responses were present in BALB/c mice rendered resistant by IL-2 neutralization and CD4+ Th2 responses in mice rendered susceptible by IL-2 treatment . The presence of IL-2 restored Ag-specific responsiveness in vitro and correlated in vivo with the expansion of CD4+ MEL-149(low) cells capable of producing IL-2 and IL-4 both in vitro and in vivo as observed in adult thymectomized mice . These results indicate that production of IL-2 early in infection correlates with the induction of IL-4-producing CD4+ Th2 cells, while a transient loss of T cell responsiveness, such as IL-2 production, appears to be required for CD4+ Th1 occurrence in mice with candidiasis.

J Biol Chem, 1997 Feb 28, 272(9), 5682 - 8
The mutation T315A in Candida albicans sterol 14alpha-demethylase causes reduced enzyme activity and fluconazole resistance through reduced affinity; Lamb DC et al.; Sterol 14alpha-demethylase (P45051) is the target for azole antifungal compounds, and resistance to these drugs and agrochemicals is of significant practical importance . We undertook site-directed mutagenesis of the Candida albicans P45051 heterologously expressed in Saccharomyces cerevisiae to probe a model structure for the enzyme . The change T315A reduced enzyme activity 2-fold as predicted for the removal of the residue that formed a hydrogen bond with the 3-OH of the sterol substrate and helped to locate it in the active site . This alteration perturbed the heme environment, causing an altered reduced carbon monoxide difference spectrum with a maximum at 445 nm . The changes also reduced the affinity of the enzyme for the azole antifungals ketoconazole and fluconazole and after expression induced by galactose caused 4-5-fold azole resistance in transformants of S . cerevisiae . This is the first example of a single base change in the target enzyme conferring resistance to azoles through reduced azole affinity.

Cell Immunol, 1997 Feb 25, 176(1), 75 - 81
Differential effects of granulocyte/macrophage colony-stimulating factor (GM-CSF) in enhancing macrophage resistance to Legionella pneumophila vs Candida albicans; Yamamoto Y et al.; It has been reported that granulocyte/macrophage colony-stimulating factor (GM-CSF), one of the hemopoietic growth factors which regulates the function of phagocytic cells, is a potent activator of cultured macrophages and induces antimicrobial activities as well as differentiation of precursor cells . In this study, we examined the ability of recombinant murine GM-CSF to activate mouse peritoneal macrophages to restrict the growth of two different microorganisms, Candida albicans and Legionella pneumophila, both of which are important opportunistic pathogens in an immunocompromised host . Treatment of thioglycollate-elicited BDF1 mouse macrophages with GM-CSF for 24 hr enhanced the anti-C . albicans activity of the macrophages in terms of inhibiting growth of the fungi . Reactive oxygen (H2O2) and IL-1 production by the macrophages were also enhanced by treatment with GM-CSF . However, no enhancement of anti-L . pneumophila activity of macrophages obtained from either susceptible A/J or resistant BDF1 mice to L . pneumophila infection after treatment with up to 1000 units/ml GM-CSF was observed under the same conditions . When the treatment time was extended to 72 hr . GM-CSF was still unable to induce anti-L . pneumophila activity . As a control study, treatment with recombinant IFN-gamma enhanced both anti-Candida and anti-Legionella activity in cultured macrophages under the same conditions used in the GM-CSF study . Measurement of cellular iron content revealed the low iron content in IFN-gamma-treated macrophages, but no decrease of iron in GM-CSF-treated macrophages compared with the control group, indicating a possible involvement of iron as a key factor in anti-L . pneumophila activity . Thus, the results of the study show that GM-CSF activation of elicited peritoneal macrophages is selective with regard to the type of antimicrobial activity induced.

FEMS Microbiol Lett, 1997 Feb 15, 147(2), 189 - 93
Reduced intracellular accumulation of azole antifungal results in resistance in Candida albicans isolate NCPF 3363; Lamb DC et al.; Candida albicans strain NCPF 3363 was isolated from a British patient with chronic mucocutaneous candidiasis (CMC) and confirmed to be resistant to azole antifungal compounds . In this study we investigate the molecular basis of resistance and show that azole tolerance in NCPF 3363 was associated with reduced intracellular accumulation of drug and not reduced affinity for the target site, as previously indicated . Relative impermeability or the presence of transporters related to those responsible for multidrug resistance are implicated in the mechanism of resistance.

J Biol Chem, 1997 Feb 7, 272(6), 3161 - 7
Identification of Glu-330 as the catalytic nucleophile of Candida albicans exo-beta-(1,3)-glucanase; Mackenzie LF et al.; The exo-beta-(1,3)-glucanase from Candida albicans hydrolyzes cell wall beta-glucans via a double-displacement mechanism involving a glycosyl enzyme intermediate . Reaction of the enzyme with 2',4'-dinitrophenyl-2-deoxy-2-fluoro-beta-D-glucopyranoside resulted in the time-dependent inactivation of this enzyme via the accumulation of a 2-deoxy-2-fluoro-glycosyl-enzyme intermediate as monitored also by electrospray mass spectrometry . The catalytic competence of this intermediate is demonstrated by its reactivation through hydrolysis (kreact = 0.0019 min-1) and by transglycosylation to benzyl thio-beta-D-glucopyranoside (kreact = 0.024 min-1; Kreact = 56 mM) . Peptic digestion of the labeled enzyme followed by tandem mass spectrometric analysis in the neutral loss mode allowed detection of two glycosylated active site peptides, the sequences of which were identified as NVAGEW and NVAGEWSAA . A crucial role for Glu-330 is confirmed by site-directed mutagenesis at this site and kinetic analysis of the resultant mutant . The activity of the Glu-330 --> Gln mutant is reduced over 50,000-fold compared to the wild type enzyme . The glutamic acid, identified in the exoglucanase as Glu-330, is completely conserved in this family of enzymes and is hereby identified as the catalytic nucleophile.

Int J Immunopharmacol, 1997 Feb, 19(2), 75 - 82
Upregulation of phagocytosis and candidicidal activity of macrophages exposed to the immunostimulant acemannan; Stuart RW et al.; Previous studies by these investigators have shown that mannosylated bovine serum albumin (m-BSA) enhances the respiratory burst (RB), phagocytosis, and killing of Candida albicans by resident murine peritoneal macrophages (MO) . Upregulation of the above MO functions was associated with binding of m-BSA to the MO-mannose receptor . The present study was done to determine if the immunostimulant, acemannan prepared from aloe vera, could stimulate MO in a similar manner . Resident peritoneal MO collected from C57BL/6 mice were exposed to acemannan for 10 min . The RB was measured using chemiluminescence and demonstrated approximately a two-fold increase above the media controls . In studies involving phagocytosis, MO were exposed to acemannan, washed and exposed to Candida at a ratio of 1:5 . The percent phagocytosis and Candida killing were determined using fluorescence microscopy . There was a marked increase in phagocytosis in the treated cultures (45%) compared to controls (25%) . Macrophages exposed to acemannan for 10 min resulted in ca 38% killing of Candida albicans compared with 0-5% killing in controls . If MO were incubated with acemannan for 60 min, 98% of the yeast were killed compared to 0-5% in the controls . The results of the present study indicate that short term exposure of MO to acemannan upregulates the RB, phagocytosis and candidicidal activity . Further studies are needed to clarify the potential use of this immunostimulant as an anti-fungal agent.

J Infect Dis, 1997 Feb, 175(2), 421 - 8
Phagocytosis of viable Candida albicans by alveolar macrophages: lack of opsonin function of surfactant protein A; Rosseau S et al.; Surfactant protein A (SP-A) contributes to host defense by opsonizing microbial organisms for phagocytosis by alveolar macrophages (AM) . The role of SP-A as opsonin for phagocytosis of Candida albicans was analyzed . AM in suspension exhibited no phagocytosis of nonopsonized yeast . This was not increased by SP-A, whether provided for preincubation of AM or yeast or present during coincubation . However, the engulfment of serum-opsonized yeast by AM in suspension was inhibited by SP-A . This inhibitory effect was mimicked by complement subcomponent C1q and concanavalin A but not by type IV collagen . SP-A did not interfere with phagocytosis of serum-opsonized yeast by adherent AM, monocytes, neutrophils, or peritoneal macrophages . SP-A lacks function as an opsonin for the phagocytosis of C . albicans by AM but interferes with binding of yeast to AM, inhibiting subsequent ingestion . The role of SP-A as an alveolar space opsonin may thus critically depend on the microbial species involved.

Hautarzt, 1997 Feb, 48(2), 94 - 9
{Characterization of nonresponders in high dosage UVA1 therapy of acute exacerbated atopic dermatitis}; Schempp CM et al.; High-dose UVA1 therapy is an effective treatment of patients with acute atopic dermatitis . However, some patients do not respond well to this new therapy . We attempted to further characterize the non-responder population in a retrospective study . Two closely matched groups of responders (n = 20) and non-responders (n = 20) were compared . No significant differences were observed between both groups with respect to the following parameters: skin type, minimal erythema dose, single and cumulative doses of UVA1, and peripheral blood eosinophils . However, non-responders were characterized by a highly elevated atopic score, and by high levels of total IgE and of specific IgE . Furthermore, colonization of the skin with Staphylococcus aureus occurred at higher densities, and intestinal growth of Candida albicans was more frequently observed . These data indicate that high-dose UVA1 irradiation is not effective in all patients suffering from atopic dermatitis . We conclude that non-responders with complicating infections might benefit from the combination of high-dose UVA1 therapy and antibiotic or antimycotic treatment.

Genitourin Med, 1997 Feb, 73(1), 39 - 43
Risk factors for sexually transmitted diseases among women attending family planning clinics in Dar-es-Salaam, Tanzania; Gertig DM et al.; BACKGROUND: Identification of risk factors for sexually transmitted diseases (STDs) assists in development of treatment algorithms, which are potentially important components of STD control when microbiologic facilities are limited . METHODS: A cross-sectional study was performed to assess STD and HIV risk factors of 2285 women attending three family planning clinics in Dar-es-Salaam, Tanzania during 1991-92 . Women were interviewed and examined for signs of STDs . Specimens were taken for laboratory diagnosis of HIV, other sexually transmitted organisms, and Candida albicans . RESULTS: The prevalence of gonorrhoea was found to be 4.2%, prevalence of trichomoniasis was 14.3%, and positive syphilis serology was found in 2.5% of women . Unmarried women were at increased risk of trichomoniasis (age-adjusted OR = 1.48 95% CI {1.12, 1.95}), gonorrhoea (age-adjusted OR = 1.81 95% CI {1.14, 2.86}) and syphilis (age-adjusted OR 1.5 {0.84, 2.68}) . An increasing number of sexual partners in the past five years was associated with an increased risk of all STDs . Current use of the oral contraceptive pill was positively associated with gonorrhoea, multivariate OR = 1.75 95% CI {1.05, 2.93} . The prevalence of candidiasis was 11.5% and was not associated with any of the demographic or behavioural risk factors examined . Clinical diagnostic algorithms for STDs in this study population had relatively low sensitivity and low positive predictive value . CONCLUSION: Being unmarried and having a higher number of sexual partners were consistently associated with each STD, while the associations for other risk factors varied between STDs, emphasising the complexity of STD distribution . Further development of diagnostic algorithms and other methods for screening women for STDs are needed to reduce the impact of STDs and HIV in developing countries.

Chem Pharm Bull (Tokyo), 1997 Feb, 45(2), 321 - 6
Optically active antifungal azoles . VII . Synthesis and antifungal activity of stereoisomers of 2-{(1R,2R)-2-(2, 4-difluorophenyl)-2-hydroxy-1-methyl-3-(1H-1,2,4-triazol-1-yl) propyl}-4-{4-(2,2,3,3-tetrafluoropropoxy)phenyl}-3(2H,4H)-1,2,4-triazo lone (TAK-187); Tasaka A et al.; 2-{(1R,2R)-2-(2,4-Difluorophenyl)-2-hydroxy-1-methyl-3-(1H-1,2, 4-triazol-1-yl)propyl}-4-{4-(2,2,3,3,tetrafluoropropoxy) phenyl}-3(2H, 4H)-1,2,4-triazolone {(1R,2R)-1: TAK-187} is a new antifungal agent selected as a candidate for clinical trials . The three stereoisomers {(1S,2S)-, (1R,2S)- and (1S,2R)-1} of this compound were prepared to clarify the relationship between the stereochemistry and the biological activities . In vitro and in vivo assays of antifungal activity revealed that TAK-187 {(1R,2R)-1} is the most potent among the four stereoisomers . Furthermore, TAK-187 was found to exert strong and selective inhibitory effect on the sterol synthesis in Candida albicans as compared with that in rat liver.

Clin Infect Dis, 1997 Feb, 24(2), 258 - 60
Myeloperoxidase deficiency manifesting as pustular candidal dermatitis; Nguyen C et al.; Myeloperoxidase deficiency is the most common neutrophilic lysosomal enzyme deficiency . Case studies indicate that individuals with myeloperoxidase deficiency are not susceptible to serious infection in the absence of coexisting conditions such as diabetes mellitus . We present a case of myeloperoxidase deficiency manifesting as disseminated pustular candidal dermatitis in a nondiabetic male . Ceftriaxone therapy was administered to the patient for 8 days after he received a closed head injury and before the development of fever and pustular dermatitis . Candida albicans was isolated from the skin lesion . His neutrophils demonstrated a qualitative lack of myeloperoxidase . Patients who develop rapidly disseminated fungal dermatitis while they are receiving antimicrobial therapy that is relatively limited in coverage should be evaluated for myeloperoxidase deficiency.

Clin Infect Dis, 1997 Feb, 24(2), 201 - 10
Vaginal colonization or infection with Candida albicans in human immunodeficiency virus-infected women during pregnancy and during the postpartum period . Women and Infants Transmission Study Group; Burns DN et al.; We evaluated the relationship between immunologic status and vaginal colonization or infection with Candida albicans for 605 women enrolled in a multicenter, prospective cohort study of mother-to-infant transmission of human immunodeficiency virus type 1 (HIV-1) . A low CD4+ lymphocyte level (< 14% vs . > or = 14%, which corresponds to an absolute count of approximately 200 x 10(6)/L) was associated with a two- to fivefold increased likelihood of vaginal colonization (odds ratio {OR}, 2.28; 95% confidence interval {CI}, 1.01-5.19) and vaginal candidiasis (OR, 3.08; 95% CI, 1.21-7.71) during pregnancy and during the postpartum period (OR, 2.98; 95% CI, 1.51-5.88 and OR, 5.45; 95% CI, 1.73-16.6, respectively) . These associations persisted in multivariate logistic regression analyses . No associations with CD8+ lymphocyte levels or CD8+ CD38+ or other lymphocyte subset levels were found after adjustment for CD4+ cell level and other covariates . However, postpartum (but not antepartum) antibiotic use and pregnancy were also associated with vaginal colonization and candidiasis (P < or = .001 for each) . Vaginal candidiasis was not associated with an increased risk of mother-to-infant transmission of HIV-1; however, a related, more inclusive variable, clinical vaginitis or vaginosis of any etiology at the last antepartum visit, was associated with mother-to-infant transmission (OR, 1.92; 95% CI, 1.07-3.43) . These findings emphasize the complex, multifactorial nature of vaginal candidiasis and highlight the need for safe and effective treatment and prevention strategies for women with advanced HIV infection.

Clin Infect Dis, 1997 Feb, 24(2), 124 - 30
Patterns of fluconazole susceptibility in isolates from human immunodeficiency virus-infected patients with oropharyngeal candidiasis due to Candida albicans; Laguna F et al.; We evaluated 119 episodes of oropharyngeal candidiasis due to C . albicans to study the patterns of fluconazole susceptibility of the isolates and the characteristics of the patients and to confirm the correlation between fluconazole susceptibility of isolates and therapeutic outcome . Sixty-one isolates were considered susceptible to fluconazole (MICs, < or = 0.5 microg/mL), 33 were intermediate (MICs, 1.0-8.0 microg/mL), and 25 were resistant (MICs, > or = 16.0 microg/mL) . Patients infected with resistant strains had significantly lower CD4+ cell counts and a less recent AIDS diagnosis than patients infected with intermediate or susceptible strains . Previous fluconazole therapy and prophylaxis were significantly more frequent for patients infected with resistant and intermediate strains (P < .001) . Decreased susceptibility to ketoconazole and itraconazole was observed in resistant and intermediate strains . Fluconazole treatment was ineffective for patients infected with resistant isolates; however, high doses of ketoconazole or itraconazole were successful for nine (81%) of them . Different patterns of fluconazole susceptibility among C . albicans strains are correlated with patients' characteristics and with therapeutic outcomes.

J Chemother, 1997 Feb, 9(1), 17 - 22
Inhibition of intracellular growth of Staphylococcus aureus by exposure of infected human monocytes to clarithromycin and azithromycin; Fietta A et al.; The direct effect of clarithromycin and azithromycin on human polymorphonuclear leukocyte (PMN) functions and their intracellular activity against Staphylococcus aureus, phagocytosed by human monocytes, were studied . The presence of both antibiotics, in the range of concentrations from 0.25 to 20 micrograms/ml, did not affect chemotaxis, opsonized-zymosan phagocytosis, respiratory burst measured by nitroblue tetrazolium reduction and phorbol myristate acetate-induced superoxide production, or the microbicidal activity of human PMNs against Candida albicans . Both macrolides were bactericidal against staphylococci in the monocyte system, while bacteriostatic activity was found in cell free system . At concentrations equal to the minimum inhibitory concentrations (MICs) (0.75 and 0.1 respectively for azithromycin and clarithromycin) more than 99% of intraphagocytic S . aureus were killed after 24 h incubation . Increasing the concentrations of each drug above the MICs (5 and 10 MICs) did not alter the killing rate of intracellular bacteria . Moreover, no differences between the intracellular bioactivity of these antibiotics were demonstrated, despite their different uptake kinetics.

Can J Microbiol, 1997 Feb, 43(2), 122 - 8
Characterization of a lipopeptide-resistant strain of Candida albicans; Frost DJ et al.; Lipopeptides are antifungal agents that inhibit cell wall beta-(1,3)-glucan biosynthesis in fungal organisms . A mutant resistant to lipopeptides was generated by UV mutagenesis and characterized . The Candida albicans mutant (LP3-1) was stable and showed resistance specificity to a broad range of lipopeptides and certain glycolipid inhibitors . Other antifungal agents with diverse modes of action had a normal minimum inhibitory concentration profile for LP3-1 compared with the wild-type strain (CCH 442) . In the in vitro beta-(1,3)-glucan synthase assay, both the lipopeptides and papulacandin-related agents had considerably higher 50% inhibitory concentration values in the LP3-1 strain than in the wild-type strain . In reconstitution assays, the resistance factor was associated with the integral membrane pellet rather than the peripheral GTP-binding protein . The LP3-1 strain had a membrane lipid profile similar to that of the parent strain and was virulent in a murine model of systemic candidiasis . Taken together, these results indicate that the resistance factor is associated with the integral membrane component of beta-(1,3)-glucan synthase . Lipopeptides are common antifungal agents encountered during screening of natural products . The LP3-1 strain was resistant to natural product extracts known to contain various lipopeptides . Thus, LP3-1 can be used in a dereplication assay.

Vaccine, 1997 Feb, 15(2), 220 - 4
Liposomes containing Candida albicans ribosomes as a prophylactic vaccine against disseminated candidiasis in mice; Eckstein M et al.; This study describes the development of prophylactic anti-Candida vaccine which combines the advantages of Candida albicans ribosomes as antigen(s) and of liposomes as carrier/adjuvant with minimal side-effects, which could be suitable for human use . The liposomal vaccine was composed of C . albicans ribosomes and the lipids dimyristoyl phosphatidyl choline (DMPC) and dimyristoyl phosphatidyl glycerol (DMPG) (9:1 molar ratio) . Some of the vaccines contained Lipid-A (LA) as an additional adjuvant . The vaccine was prepared by two methods: (I) dehydration by lyophylization of small liposomes in the presence of the Candida ribosomes (DRV method); (II) colyophylization of DMPC/DMPG (9:1 mole ratio) in tertiary butanol with aqueous dispersion of Candida ribosomes . In both cases a dry powder was obtained which was rehydrated to form large multilamellar vesicles . The efficacy of the vaccines in mice was tested by their protectivity against a challenge with C . albicans, as assessed by survival rate, induction of cell mediated immunity (measured by delayed type hypersensitivity-DTH) and anti-Candida antibody titer . Unimmunized mice and mice vaccinated by ribosomes supplemented with incomplete Freund's adjuvant (IFA) were used as controls . The results indicated that the liposomal vaccines were effective at least as the IFA-based vaccine . The study indicates the feasibility of developing an efficacious anti-Candida vaccine for human use.

Thorax, 1997 Feb, 52(2), 161 - 5
Exercise induced bronchospasm in Ghana: differences in prevalence between urban and rural schoolchildren; Addo Yobo EO et al.; BACKGROUND: As more developing countries adopt a westernised style of living, an increase in the prevalence of asthma can be expected to occur in these areas . A study was undertaken to establish the normal response to exercise in Ghanaian children and to use these normal values to determine the prevalence of exercise induced bronchospasm (EIB) in urban rich (UR), urban poor (UP), and rural (R) school children . Skin test reactivity to common inhalant allergens in UR, UP, and R children with and without EIB was also investigated . METHODS: Two hundred children aged 9-16 years without a previous history of respiratory symptoms were randomly selected and underwent free running exercise testing . A normal response to exercise was defined as the group mean change in peak expiratory flow rate (PEFR) +/- 2 standard deviations . This value was used to identify the prevalence of EIB in UR, UP, and R schoolchildren . A total of 1095 children from three different schools underwent exercise testing (220 UP, 599 UR, 276 R), after which 916 children underwent skin prick testing to six common inhalant allergens (D farinae, D pteronyssinus, cat, dog, Aspergillus flavus and Candida albicans) . RESULTS: From the results of exercise testing in asymptomatic children the normal range was defined as a fall in PEFR of < 12.5% after exercise . Thirty four children were classified as having EIB on the basis of the above definition, giving an overall prevalence of 3.1% . The prevalence of EIB was significantly higher in UR children (4.7%) than in both UP (2.2%; p < 0.05) and R children (1.4%; p < 0.01) . However, the prevalence rates in the UP and R children were similar . The prevalence of atopy in the whole population was 4.4% . Of the children with EIB, 10% were skin test positive to at least one of the allergens tested . The prevalence of atopy was significantly higher in UR children (6.55%, 95% confidence interval (CI) 4.5% to 9.2%) than in UP (2.9%, 95% CI 0.9% to 6.7%) and R children (1.5%, 95% CI 0.4% to 3.7%), respectively (p < 0.005) . CONCLUSIONS: The prevalence of EIB and atopy is higher in urban rich than in urban poor or rural children suggesting that, in addition to genetic predisposition, social and environmental factors such as wealth, life style, and housing are important determinants of these phenotypes.

Microbiology, 1997 Feb, 143 ( Pt 2), 437 - 48
Metabolism of inositol 1,4,5-trisphosphate in Candida albicans: significance as a precursor of inositol polyphosphates and in signal transduction during the dimorphic transition from yeast cells to germ tubes; Gadd GM et al.; The metabolism of inositol 1,4,5-trisphosphate {Ins(1,4,5)P3} was examined in yeast cells and germ tubes of Candida albicans . Methods have been developed for analysis of the two key metabolic enzymes, Ins(1,4,5)P3, kinase and phosphatase . ATP-dependent Ins(1,4,5)P3 kinase activity was detected predominantly in the soluble fraction of cell extracts and exhibited a Km of approximately 9 microM . The apparent Km of Ins(1,4,5)P3 phosphatase for Ins(1,4,5)P3 was approximately 480 microM . The slow rate of dephosphorylation of Ins(1,4,5_P3 to inositol bisphosphate suggests a lower importance of the phosphatase within cells compared to the kinase . Since both yeast cells and germ tubes of C . albicans rapidly phosphorylated Ins(1,4,5)P3 to inositol tetrakisphosphate and inositol penta/hexakisphosphate, it is suggested that Ins(1,4,5)P3 has an important role as a precursor for production of these compounds . A sustained increase in cellular Ins(1,4,5)P3 levels was observed during germ tube formation and, prior to the onset of germination between 1 and 2 incubation, the Ins(1,4,5)P3 content increased up to eightfold . Transient increases in the level of Ins(1,4,5)P3 were also observed during yeast-like growth of C . albicans . The possible role and relative importance of Ins(1,4,5)P3 as a precursor for inositol polyphosphates and in signal transduction involving Ca2+ release from internal stores is discussed.

Microbiology, 1997 Feb, 143 ( Pt 2), 429 - 35
Biochemical and genetic characterization of Rbf1p, a putative transcription factor of Candida albicans; Ishii N et al.; A Candida albicans gene encoding a novel DNA-binding protein that bound to the RPG box of Saccharomyces cerevisiae and the telomeric repeat sequence of C . albicans was previously cloned and designated RBF1 (RPG-box-binding factor) . In this report, determination of the functional domains of the protein is described . The DNA-binding domain was 140 aa in length, was centrally located between two glutamine-rich regions, and correlated with transcriptional activation in S . cerevisiae . The results, together with the previous finding that showed its predominant localization in the nucleus, suggest that this DNA-binding protein could be a transcription factor . Disruption of the functional RBF1 gene of C . albicans strains caused an alteration in cell morphology to the filamentous form on all solid and liquid media tested . Thus, we speculate that Rbf1p may be involved in the regulation of the transition between yeast and filamentous forms at the level of transcription.

Microbiology, 1997 Feb, 143 ( Pt 2), 417 - 27
A DNA-binding protein from Candida albicans that binds to the RPG box of Saccharomyces cerevisiae and the telomeric repeat sequence of C . albicans; Ishii N et al.; Electromobility shift assays with a DNA probe containing the Saccharomyces cerevisiae ENO1 RPG box identified a specific DNA-binding protein in total protein extracts of Candida albicans . The protein, named Rbf1p (RPG-box-binding protein 1), bound to other S . cerevisiae RPG boxes, although the nucleotide recognition profile was not completely the same as that of S . cerevisiae Rap 1p (repressor-activator protein 1), an RPG-box-binding protein . The repetitive sequence of the C . albicans chromosomal telomere also competed with RPG-box binding to Rbf1p . For further analysis, we purified Rbf1p 57,600-fold from C . albicans total protein extracts, raised mAbs against the purified protein and immunologically cloned the gene, whose ORF specified a protein of 527 aa . The bacterially expressed protein showed RPG-box-binding activity with the same profile as that of the purified one . The Rbf1p, containing two glutamine-rich regions that are found in many transcription factors, showed transcriptional activation capability in S . cerevisiae and was predominantly observed in nuclei . These results suggest that Rbf1p is a transcription factor with telomere-binding activity in C . albicans.

Microbiology, 1997 Feb, 143 ( Pt 2), 405 - 16
Cloning of Candida albicans genes conferring resistance to azole antifungal agents: characterization of CDR2, a new multidrug ABC transporter gene; Sanglard D et al.; Resistance to azole antifungal agents in Candida albicans can be mediated by multidrug efflux transporters . In a previous study, we identified at least two such transporters, Cdr1p and Benp, which belong to the class of ATP-binding cassette (ABC) transporters and of major facilitators, respectively . To isolate additional factors potentially responsible for resistance to azole antifungal agents in C . albicans, the hypersusceptibility of a Saccharomyces cerevisiae multidrug transporter mutant, delta pdr5, to these agents was complemented with a C . albicans genomic library . Several new genes were isolated, one of which was a new ABC transporter gene called CDR2 (Candida drug resistance) . The protein Cdr2p encoded by this gene exhibited 84% identity with Cdr1p and could confer resistance to azole antifungal agents, to other antifungals (terbinafine, amorolfine) and to a variety of metabolic inhibitors . The disruption of CDR2 in the C . albicans strain CAF4-2 did not render cells more susceptible to these substances . When the disruption of CDR2 was performed in the background of a mutant in which CDR1 was deleted, the resulting double delta cdr1 delta cdr2 mutant was more susceptible to these agents than the single delta cdr1 mutant . The absence of hypersusceptibility of the single delta cdr2 mutant could be explained by the absence of CDR2 mRNA in azole-susceptible C . albicans strains . CDR2 was overexpressed, however, in clinical C . albicans isolates resistant to azole antifungal agents as described previously for CDR1, but to levels exceeding or equal to those reached by CDR1 . Interestingly, CDR2 expression was restored in delta cdr1 mutants reverting spontaneously to wild-type levels of susceptibility to azole antifungal agents . These data demonstrate that CDR2 plays an important role in mediating the resistance of C . albicans to azole antifungal agents.

Microbiology, 1997 Feb, 143 ( Pt 2), 397 - 404
Functional reconstitution of a purified proline permease from Candida albicans: interaction with the antifungal cispentacin; Jethwaney D et al.; We have purified proline permease to homogeneity from Candida albicans using an L-proline-linked agarose matrix as an affinity column . The eluted protein produced two bands of 64 and 67 kDa by SDS-PAGE, whereas it produced a single band of 67 kDa by native PAGE and Western blotting . The apparent Km for L-proline binding to the purified protein was 153 microM . The purified permease was reconstituted into proteoliposomes and its functionality was tested by imposing a valinomycin-induced membrane potential . The main features of L-proline transport in reconstituted systems, viz . specificity and sensitivity to N-ethylmaleimide, were very similar to those of intact cells, The antifungal cispentacin, which enters C . albicans cells via an inducible proline permease, competitively inhibited the L-proline binding and translocation in reconstituted proteoliposomes . However, the uptake of L-proline in proteoliposomes reconstituted with the purified protein displayed monophasic kinetics with an apparent Km of 40 microM.

Microbiology, 1997 Feb, 143 ( Pt 2), 377 - 86
The topoisomerase I gene from Candida albicans; Jiang W et al.; We report here the cloning of the Candida albicans genomic topoisomerase I gene (TOP1) by use of PCR and subsequent hybridization . The predicted protein sequence shared 58.8% identity with the Saccharomyces cerevisiae topoisomerase I and 30-50% identity with other eukaryotic topoisomerase I proteins . A conditional gene disruption strain (CWJ477) was constructed so that one copy of TOP1 was deleted and the other copy of TOP1 was placed under a regulatable promoter . Under repressed conditions, cells grew slowly and cell morphology was abnormal . The virulence of CWJ477 was markedly reduced in a mouse model system, and that of the single gene knockout-strain was slightly attenuated, indicating that TOP1 might play a role in the infection of C . albicans in mice in a dose-dependent manner . Despite the reduced virulence of both the single and double knockout strains, viable cells of the pathogen were recovered from the kidneys as late as 22 d post-infection.

Microbiology, 1997 Feb, 143 ( Pt 2), 367 - 76
Phenotype in Candida albicans of a disruption of the BGL2 gene encoding a 1,3-beta-glucosyltransferase; Sarthy AV et al.; The BGL2 gene encodes a unique 1,3-beta-glucosyltransferase (Bgl2p) present in the cell wall of Candida albicans and other fungi . Although believed to be involved in cell wall assembly, disruption of the gene in saccharomyces cerevisiae showed no apparent phenotype . We performed sequential disruptions of the BGL2 loci in a homozygous ura3 clinical isolate of C . albicans using the URA3 blaster method, in order to investigate the role of Bgl2p in this dimorphic, pathogenic fungus . Strain CACW-1 contained disruptions of both homologues of the BGL2 gene and lacked Bgl2p, as assessed by protein extraction, SDS-PAGE and Western blot analysis, and enzyme assay; however, residual non-Bgl2p transferase activity was detected . CACW-1 was attenuated in virulence for mice when compared to an isogenic parent strain, and fewer organisms were recovered from the kidneys of infected animals . Additional phenotypic changes included: (1) a dramatic increase in the sensitivity to the chitin synthesis inhibitor nikkomycin Z when CACW-1 cells were incubated at 37 or 42 degrees C; (2) an 8.7 +/- 1.6% slower growth rate at 37 degrees C for CACW-1 when compared to its isogenic parent; and (3) aggregation of CACW-1 cells during stationary phase and/or incubation of stationary phase cells in phosphate buffer . Characterization of SDS-extracted cell walls did not reveal any significant differences in the levels of 1,3-beta- or 1,6-beta-glucan . These data reveal that loss of Bgl2p does have a phenotype in C . albicans, and indicate that (1) loss of Bgl2p function renders cells more dependent on chitin for wall integrity, and attenuates virulence (probably due to subtle changes in wall structure), and (2) that additional 1,3-beta-glucosyltransferases are present in the C . albicans BGL2 disruptant.

Microbiology, 1997 Feb, 143 ( Pt 2), 357 - 66
N-myristoylation of Arf proteins in Candida albicans: an in vivo assay for evaluating antifungal inhibitors of myristoyl-CoA: protein N-myristoyltransferase; Lodge JK et al.; Myristoyl-CoA: protein N-myristoyltransferase (Nmt) catalyses the covalent attachment of myristate to the N-terminal glycine of a small subset of cellular proteins produced during vegetative growth of Candida albicans . nmt447D is a mutant NMT allele encoding an enzyme with a Gly447-->ASP substitution and reduced affinity for myristoyl-CoA . Among isogenic NMT/NMT, NMT/ delta nmt and nmt delta/nmt447D strains, only nmt delta/nmt447D cells require myristate for growth on yeast/peptone/dextrose media (YPD) at 24 or 37 degrees C . When switched from YPD/myristate to YPD alone, 60% of the organisms die with 4 h . Antibodies raised against the C-terminal eight residues of Saccharomyces cerevisiae Arf1p were used to probe Western blots of total cellular proteins prepared from these isogenic Candida strains . N-Myristoylation of C . albicans ADP-ribosylation factor (Arf) produced a change in its electrophoretic mobility during SDS-PAGE: the myristoylated species migrated more rapidly than the nonmyristoylated species . In an NMT/nmt delta strain, 100% of the Arf is N-myristoylated based on this mobility shift assay . When exponentially growing nmt delta/nmt447D cells were incubated at 24 degrees C in YPD/myristate, < 25% cellular Arf was nonmyristoylated . In contrast, 2 or 4 h after withdrawal of myristate, > or = 50% of total cellular Arf was nonmyristoylated . This finding suggests that > or = 50% reduction in Arf N-myristoylation is a biochemical marker of a growth-arrested cell . A similar conclusion was made after assaying isogenic S . cerevisiae strains containing various combinations of NMT1, nmt1-451D, ARF1, arf1 delta, ARF2 and arf2 delta alleles and grown at 24-37 degrees C on YPD of YPD/myristate . Peptidomimetic inhibitors of C . albicans Nmt were synthesized based on the N-terminal sequence of an S . cerevisiae Aft . SC-59383 has an IC50 of 1.45 +/- 0.08 microM for purified C . albicans Nmt and is 560-fold selective for the fungal compared to human N-myristoyltransferase . It had an EC50 of 51 +/- 17 and 67 +/- 6 microM, 24 and 48 h after a single administration of the drug to cultures of C . albicans . The Arf gel mobility shift assay indicated that a single dose of 200 microM produced a < 50% reduction in Arf N-myristoylation after 4 h, which is consistent with the fungistatic, but not fungicidal, activity . The effect on Nmt was specific: an enantiomer, SC-59840, had no inhibitory effect on purified C . albicans Nmt (IC50 > 1,000 microM), and 200 microM of the compound produced no detectable reduction in Arf N-myristoylation in vivo . SC-58272, which is related to SC-59383, was a more potent inhibitor in vitro (IC50 0.056 +/- 0.01 microM), but had no growth inhibitory activity and did not produce any detectable reduction in Arf N-myristoylation . These findings highlight the utility of the Arf protein gel mobility shift assay for demonstrating the mechanism-based antifungal activity of SC-59383, a selective inhibitor of C . albicans Nmt.

Microbiology, 1997 Feb, 143 ( Pt 2), 349 - 56
Analysis of secreted aspartic proteinases from Candida albicans: purification and characterization of individual Sap1, Sap2 and Sap3 isoenzymes; Smolenski G et al.; The recently discovered secreted aspartic proteinase multi-gene (SAP) family in Candida albicans has complicated assessment of proteolytic activity as a factor in the onset and development of Candida infections . Differential expression of the SAP genes under various conditions, as well as possible variation in the properties of the individual isoenzymes, have consequences for immunological detection, for targeted drug design and possibly for pathogenicity . It is therefore important to be able to monitor Sap isoenzyme profiles in different strains of C . albicans cultures, and to know the biochemical properties of each isoenzyme . We have employed a simple purification protocol based on strong anion exchange chromatography for the direct analysis of C . albicans Sap isoenzymes from culture filtrates, as well as recovery of individual Sap1, Sap2 and Sap3 products . In the case of Sap1, this involved development of an overexpression system using the pEMBLyex4 vector transformed into Saccharomyces cerevisiae . The C . albicans strains ATCC 10231 and 10261 were shown to produce different ratios of Sap2 and Sap3 under the same conditions . Analysis of all three purified proteins by gel electrophoresis, immunoblotting and proteinase assays which were designed to evaluate pH dependence, thermal stability and substrate specificity revealed similar but distinct properties for each isoenzyme . Although Sap3 was shown to be antigenically more similar to Sap2 than was Sap1, it was less similar in terms of thermal stability and activity at low pH, being more stable and more active.

Microbiology, 1997 Feb, 143 ( Pt 2), 341 - 8
Identification of salivary basic proline-rich proteins as receptors for Candida albicans adhesion; O'Sullivan JM et al.; The adherence of Candida albicans cells to oral surfaces is believed to be an important step in the development of oral candidosis . Electrophoretically separated parotid salivary proteins were transferred to nitrocellulose membranes and incubated with {35S}methionine-radiolabelled C . albicans cells in a cell overlay adherence assay . A subset of four proteins with apparent molecular masses of 17, 20, 24 and 27 kDa (designated bands A-D) acted as receptors for cells of C . albicans ATCC 10261 and four clinical C . albicans isolates, in overlay assays . The N-terminal amino acid sequence of bands A-D indicated that these proteins were members of the basic proline-rich protein (bPRP) family . Digestion of protein A with endoproteinase Glu-C resulted in a single band (designated Ap) detected by Coomassie blue staining after SDS-PAGE . This band was not bound by C . albicans cells in overlay assays and comprised two fragments, designated ApN and ApC . These fragments had N-terminal sequences corresponding to the N-terminal and post endoproteinase Glu-C cleavage site sequences of bPRP IB-6 and had molecular masses of 6189 and 4261 Da as determined by mass spectrometry . Thus intact bPRP IB-6, and other bPRPs, may act as receptors for C . albicans adhesion.

Microbiology, 1997 Feb, 143 ( Pt 2), 313 - 20
The highly immunogenic enolase and Hsp70p are adventitious Candida albicans cell wall proteins; Eroles P et al.; Screening cDNA libraries with polyclonal antibody preparations against Candida albicans yeast or mycelial cell walls resulted in isolation of several positive clones . Some of them encoded enolase; others encoded a protein of the 70 kDa heat-shock protein family (Hsp70p), etc . The presence of these cytosolic proteins in the cell wall of actively growing C . albicans was discovered by analytical (SDS-PAGE and Western blot) and cytological (indirect immunofluorescence) experiments . Supplementation of cell cultures with papulacandin B, an antibiotic that inhibits formation of the beta-glucan skeleton, resulted in the release of enolase to the supernatant fluids; this release was prevented when 0.6 M KCl was present as an osmotic stabilizer . The cell wall of C . albicans incorporated exogenously added proteins (enolase and Escherichia coli and C., albicans cytosolic proteins) . The presence in the C . albicans cell wall of enolase, Hsp70p, and probably other intracellular proteins that are highly immunogenic might help the fungal cells to evade the host defences, and consequently could represent a survival mechanism for C . albicans 'in vivo'.

Microbiology, 1997 Feb, 143 ( Pt 2), 303 - 11
Yeast-enhanced green fluorescent protein (yEGFP)a reporter of gene expression in Candida albicans; Cormack BP et al.; The green fluorescent protein (GFP) of Aequorea victoria has been developed here as a reporter for gene expression and protein localization in Candida albicans . When wild-type (wt) GFP was expressed in C . albicans, it was not possible to detect fluorescence or a translation product for the wt protein . Since this was probably due in part to the presence of the non-canonical CTG serine codon in the Aequorea sequence, this codon was changed to the leucine codon TTG . C . albicans cells expressing this construct contained GFP mRNA but were non-fluorescent and contained no detectable translation product . Hence a codon-optimized GFP gene was constructed in which all of the 239 amino acids are encoded by optimal codons for C . albicans . In this gene were also incorporated two previously identified mutations in the chromophore that increase GFP fluorescence . C . albicans cells expressing this yeast-enhanced GFP gene (yEGFP3) are fluorescent and contain GFP protein . yEGFP3 can be used as a versatile reporter of gene expression in C . albicans and Saccharomyces cerevisiae and the optimized GFP described here should have broad applications in these and other fungal species.

Microbiology, 1997 Feb, 143 ( Pt 2), 297 - 302
Cloning, analysis and one-step disruption of the ARG5,6 gene of Candida albicans; Negredo A et al.; The ARG5,6 gene from the dimorphic fungus Candida albicans was cloned by functional complementation of the arginine auxotrophy present in strain EL2 (Arg-) using a gene library constructed in the double autonomously replicating sequence vector pRM1 . Sequence analysis revealed a putative 857 amino acid polypeptide (95 kDa) which showed high homology (63% protein identity) to the Saccharomyces cerevisiae ARG5,6 gene . Similarly to the S . cerevisiae gene, the C . albicans ARG5,6 gene is responsible for both the acetylglutamate kinase and acetylglutamyl-phosphate reductase activities, the second and third steps of arginine biosynthesis at the mitochondria . The C . albicans ARG5,6 gene complemented the arg6 mutation present in S . cerevisiae (strain D160-4D) on a yeast episomal plasmid using its own regulatory signals . A set of non-integrative high-efficiency plasmid vectors based on this gene marker was constructed and a null C . albicans arg5,6 delta strain was obtained using the common URA3-blaster strategy . In addition, we generated an arg5,6 delta null mutant in a single transformation event, thus improving the basic strategy for generating gene deletions in C . albicans.

Microbiology, 1997 Feb, 143 ( Pt 2), 289 - 95
WO-2, a stable aneuploid derivative of Candida albicans strain WO-1, can switch from white to opaque and form hyphae; Magee BB et al.; Candida albicans strain WO-2 was isolated as a spontaneous derivative of the white-opaque switching strain WO-1 . The electrophoretic karyotype of WO-2 lacks two bands which are found in the parent . These bands correspond to one homologue of chromosome 7 and to a translocation product containing parts of chromosomes 6 and 5 . Probing a blot of the karyotype demonstrated that the genetic material in these bands had been lost, yielding an aneuploid strain . UV-irradiation experiments showed that auxotrophs due to mutation in genes located in this region predominated, supporting the conclusion that WO-2 is partially haploid . WO-2 contained about 10% of its genome in the haploid state, and it grew with a doubling time of about twice that of its parent . However, it was able to undergo both the yeast-to-hyphal transition and the white-opaque transition . Hence, these processes do not require perfect diploidy.

South Med J, 1997 Feb, 90(2), 246 - 8
Neonatal Torulopsis glabrata fungemia; Reich JD et al.; Torulopsis glabrata is a yeastlike fungus that has recently become recognized as an important opportunistic pathogen . Only four cases of T glabrata infection in neonates have been reported . We report two cases of fungemia caused by this organism in premature infants . Both patients were treated with amphotericin B and survived the fungemia, but one patient later died of bacterial sepsis . Both patients had been treated with surfactant, artificial ventilation, intravascular catheters (arterial and venous), broad spectrum antibiotics, and hyperalimentation, which appear to be risk factors for T glabrata fungemia . A review of the literature indicates that T glabrata is susceptible to amphotericin B and 5-fluorocytosine and is resistant to fluconazole . In addition, it is less susceptible to ketoconazole, clotrimazole, and itraconazole than is Candida albicans . We recommend that T glabrata infections be treated initially by reducing iatrogenic risk factors and beginning amphotericin B therapy . If necessary, 5-fluorocytosine should be added to the drug regimen.

Phytochemistry, 1997 Feb, 44(4), 575 - 9
Protective effect of iridals from saponin injury in Candida albicans cells; Leconte O et al.; Spirostanol saponin permeabilization of Candida albicans cells resulted in phosphate (Pi) leakage from these cells . Pre-incubation of C . albicans suspension cultures with ergosterol or cycloiridals inhibited Pi leakage when cells were subjected to saponins . Saponins were shown to precipitate ergosterol but not cycloiridals . Inhibition of Pi leakage in the presence of cycloiridals could thus be due to their incorporation into cell membranes . Cycloiridals, which were previously shown to modify fluidity parameters in artificial phospholipid bilayers, seem in vivo to have a protective effect on membranes against surfactant (saponin) injury.

Infect Immun, 1997 Feb, 65(2), 833 - 7
Reduced virulence of Candida albicans MKC1 mutants: a role for mitogen-activated protein kinase in pathogenesis; Diez-Orejas R et al.; Deletion of the Candida albicans mitogen-activated protein kinase MKC1 gene gave rise to viable cells whose cell integrity was affected (F . Navarro-Garcia, M . Sanchez, J . Pla, and C . Nombela, Mol . Cell . Biol . 15:2197-2206, 1995) . In an experimental infection system using a murine model, the C . albicans mkc1 delta/mkc1 delta strain was found to be less pathogenic than the parental strain, as show the different time of survival, percentage of mortality, fungal load in the most representative organs, and histological analysis . This is the first study that shows the involvement of the cell integrity pathway in the pathogenicity of a dimorphic fungus.

Infect Immun, 1997 Feb, 65(2), 829 - 32
Avirulence of Candida albicans FAS2 mutants in a mouse model of systemic candidiasis; Zhao XJ et al.; Disruption of both alleles of the Candida albicans FAS2 gene abolishes the ability of the organism to establish infection in a murine model of systemic candidiasis . Within 72 h all mice inoculated with 10(6) CFU of the parental C . albicans strain had died . In contrast, all animals inoculated with the mutant strain CFD2 survived for the course of the experiment (21 days) . Animals infected with either mutant strain CFD1 or CFD3, in which only one FAS2 allele was disrupted, also succumbed to infection, but mortality was not observed until 4 days postinfection and survivors remained for up to 20 days postinfection . The results demonstrate that FAS2 is required for successful C . albicans infection.

Infect Immun, 1997 Feb, 65(2), 661 - 7
Oral carriage of Candida albicans in murine AIDS; Deslauriers N et al.; Oral candidiasis is a common fungal infection in patients infected with the human immunodeficiency virus (HIV) . Although rare at the time of primary HIV infection, it is frequently found throughout the asymptomatic phase and is predictive of progressive immunodeficiency . However, the precise immune defect which results in outgrowth of commensal Candida albicans in HIV infection has not been identified . Mice infected with the Du5H(G6T2) mixture of mouse leukemia viruses develop a syndrome, designated murine AIDS (MAIDS), that has many of the immune abnormalities found in HIV infection . Retrovirus-infected C57BL/6 mice were examined for their ability to resist the development of oral candidiasis from the carrier state established after a self-limiting acute infection and to clear a subsequent secondary inoculum of oral C . albicans . Most of the mice orally colonized with C . albicans and then inoculated with the retrovirus mixture maintained a low-level oral carriage of C . albicans, while 30% of coinfected mice developed recurring 2- to 3-week episodes of acute Candida proliferation, separated by transient recoveries to the carrier state . The frequencies of CD4+ and CD8+ lymphocytes were, respectively, unchanged and significantly decreased (P < 0.05) in both cervical lymph nodes and spleens of coinfected mice compared to the corresponding frequencies in C . albicans-carrying, virus-free, age-matched control animals . Secretion of gamma interferon by concanavalin A (ConA)-stimulated spleen cells from Candida-carrying, retrovirus-infected mice was significantly decreased (P < 0.05) compared to that of C . albicans-carrying, retrovirus-free mice, in accordance with known abnormalities associated with MAIDS . However, production of this cytokine by ConA-stimulated or unstimulated cervical lymph node cells from coinfected mice was enhanced compared to that of virus-free animals colonized with C . albicans . Acquired resistance to reinfection with C . albicans was maintained in retrovirus-infected mice and was associated with a mucosal recruitment of CD8+ cells not observed in control mice . These results suggest that alterations in mucosal immunity which occur in MAIDS differ substantially from defects observed at other sites and that surrogate epithelial defense mechanisms may function locally to limit Candida proliferation.

Infect Immun, 1997 Feb, 65(2), 551 - 6
Role of aspartic proteases in disseminated Candida albicans infection in mice; Fallon K et al.; A murine model of disseminated candidiasis involving intranasal challenge with Candida albicans was developed and used to explore the role of C . albicans aspartic proteases as virulence factors during early dissemination . Pretreatment of neutropenic mice with the aspartic protease inhibitor pepstatin A by intraperitoneal injection afforded strong dose-dependent protection against a subsequent lethal intranasal dose of an aspartic protease-producing strain (ATCC 32354) of C . albicans . Administration of 0.6 mg of pepstatin A kg of body weight(-1) prior to challenge and on days 1 to 4 postchallenge resulted in 100% survival at day 15 postchallenge, whereas 100% of animals receiving saline had died by day 6 . This effect was comparable to the dose-dependent protection obtained with amphotericin B, which resulted in 100% survival when administered at 0.1 mg kg(-1) . The reduction in mortality afforded by pepstatin A correlated with its dose-dependent blockade of C . albicans numbers in the lungs, liver, and kidneys . By sharp contrast, no protection by pepstatin A was observed in mice challenged intravenously, and protection was markedly attenuated in mice given pepstatin A after intranasal challenge only . These data show the utility of pepstatin A in the prophylaxis of disseminated Candida infections and suggest that Candida aspartic proteases play an essential role early in dissemination.

J Clin Microbiol, 1997 Feb, 35(2), 358 - 63
Rapid susceptibility testing of Candida albicans by flow cytometry; Kirk SM et al.; The emerging magnitude of human fungal infections has renewed interest in developing rapid and standardized methods for susceptibility testing . We demonstrated that susceptibility testing of Candida albicans can be accomplished rapidly by using flow cytometry . Test results were available within 8 to 24 h after C . albicans isolates were incubated with amphotericin B, itraconazole, and flucytosine . This is an improvement of 24 to 60 h in the time to availability of susceptibility test results compared to the time to availability of National Committee for Clinical Laboratory Standards-recommended broth macrodilution test results . In addition, the flow cytometric endpoints, mean channel fluorescence, and number of fluorescence-labeled C . albicans cells were easy to interpret for greater sensitivity and reliability . Flow cytometry provides a more accurate means of obtaining antifungal susceptibility test results.

Mol Gen Genet, 1997 Jan 27, 253(4), 520 - 8
The Aspergillus nidulans genes chsA and chsD encode chitin synthases which have redundant functions in conidia formation {corrected and republished article originally appeared in Mol Gen Genet 1996 Jun; 251(4):442-50}; Motoyama T et al.; We previously isolated three chitin synthase genes (chsA, chsB, and chsC) from Aspergillus nidulans . In the present work, we describe the isolation and characterization of another chitin synthase gene, named chsD, from A . nidulans . Its deduced amino acid sequence shows 56.7% and 55.9% amino acid identity, respectively, with Cal1 of Saccharomyces cerevisiae and Chs3 of Candida albicans . Disruption of chsD caused no defect in cell growth or morphology during the asexual cycle and caused no decrease in chitin content in hyphae . However, double disruption of chsA and chsD caused a remarkable decrease in the efficiency of conidia formation, while double disruption of chsC and chsD caused no defect . Thus it appears that chsA and chsD serve redundant functions in conidia formation.

J Med Chem, 1997 Jan 17, 40(2), 201 - 9
Synthesis and inhibition studies of sulfur-substituted squalene oxide analogues as mechanism-based inhibitors of 2,3-oxidosqualene-lanosterol cyclase; Stach D et al.; The synthesis and biological evaluation of three new sulfur-substituted oxidosqualene (OS) analogues (1-3) are presented . In these analogues, C-11, C-15, or C-18 in the OS skeleton was replaced by sulfur . The sulfur position in the OS skeleton was chosen to disrupt one or more key processes involved in cyclization: (a) the folding of the B-ring into a boat conformation, (b) the anti-Markovnikov cyclization leading to the C-ring, or (c) the formation of the D-ring during the lanosterol biosynthesis . Enzyme inhibition kinetics using homogeneous mammalian oxidosqualene cyclases (OSC) were also examined for the previously reported S-19 analogue 4 . The four analogues were potent inhibitors of mammalian OSCs (IC50 = 0.05-2.3 microM for pig and rat liver OSC) and fungal cell-free Candida albicans OSC (submicromolar IC50 values) . In particular, the S-18 analogue 3 showed the most potent inhibition toward the rat liver enzyme (IC50 = 50 nM) and showed potent, selective inhibition against the fungal enzyme (IC50 = 0.22 nM, 10-fold more potent than the S-19 analogue 4) . Thus, 3 is the most potent OSC inhibitor known to date . The Ki values ranged from 0.5 to 4.5 microM for pig OSC, with 3 and 4 showing about 10-fold higher potency for rat liver OSC . Interestingly, the S-18 analogue 3 showed time-dependent irreversible inhibition with homogeneous pig liver OSC (kinact = 0.06 min-1) but not with rat OSC.

Nucleic Acids Res, 1997 Jan 15, 25(2), 431 - 7
Expression of a reporter gene interrupted by the Candida albicans group I intron is inhibited by base analogs; Mercure S et al.; We previously reported the identification of an intron (CaLSU) in the 25S ribosomal RNA of some Candida albicans yeast strains . CaLSU was shown to self-splice and has the potential to adopt a secondary structure typical of group I introns . The presence of CaLSU inC . albicans strains correlates with a high degree of susceptibility to base analog antifungal agents, 5-fluorocytosine (5-FC) or 5-fluorouracil (5-FU) . Cell death, resulting from addition of base analogs to growing cultures, precluded demonstration of a causal relationship between CaLSU presence and susceptibility to base analogs . In the present study, CaLSU was inserted in a non-essential lacZ reporter gene and expression was examined in Saccharomyces cerevisiae . Different mutations affecting in vitro self-splicing also had similar effects on reporter gene expression in vivo . This indicates that in vivo removal of CaLSU from the reporter gene occurs through the typical self-splicing mechanism of group I introns . Base analogs inhibited expression of the reporter gene product in a concentration-dependent manner upon their addition to the cultures . This supports a model in which disruption of intron secondary structure, consecutive to the incorporation of nucleotide analogs, is a major factor determining the susceptibility of C.albicans cells to base analogs.

FEMS Microbiol Lett, 1997 Jan 15, 146(2), 255 - 61
Partial purification of (1,3)-beta-glucan synthase from Candida albicans; Frost D et al.; (1,3)-beta-Glucan synthase from Candida albicans was solubilized from microsomal membranes using the detergent 3-{(3-cholamidopropyl) dimethylammonio}-1-propane sulfonate (Chaps) . Effective solubilization was dependent upon the strain and the method used to detect enzyme activity . The solubilized enzyme was purified over 765-fold using a modified product entrapment technique . Bovine serum albumin, an activator of glucan synthase, precipitated proteins during product entrapment and was replaced with BSA immobilized on agarose beads . SDS-PAGE analysis revealed a prominent 187-kDa band present in the product entrapped pellet as well as several additional polypeptides at 227, and 187, 182 and 39 kDa which were not prevalent in crude preparations.

FEBS Lett, 1997 Jan 2, 400(1), 80 - 2
Resistance to fluconazole and cross-resistance to amphotericin B in Candida albicans from AIDS patients caused by defective sterol delta5,6-desaturation; Kelly SL et al.; Fluconazole resistance occurs in > 10% of cases of candidosis during the late stages of AIDS . We show here in two clinical isolates that resistance was caused by defective sterol delta5,6-desaturation . This altered the type of sterol accumulating under fluconazole treatment from 14alpha-methylergosta-8,24(28)-dien-3beta,6alpha -diol to 14alpha-methylfecosterol which is capable of supporting growth . A consequence of this mechanism of azole resistance is that an absence of ergosterol causes cross-resistance to the other major antifungal agent available, amphotericin B . The results also show that growth arrest after fluconazole treatment of C . albicans in clinical conditions is caused by 14alpha-methylergosta-8,24(28)-dien-3beta,6alpha -diol accumulation.

Akush Ginekol (Sofiia), 1997, 36(1), 32 - 3
{The treatment of acute mycotic colpitis with ciclopiroxolamine in patients using oral contraceptives}; Damianov L et al.; The author reveal the results of treatment with ciclopiroxolamine of women suffering from mycotic colpitis and using OC's . The research covers 26 women users of monophasic OC's with mycotic colpitis during the last 2-3 weeks before the research . The cause for the colpitis in 92% of cases is Candida albicans . The treatment was successful in 84.6% of the cases after the first 6-day-course with Batrafen creme . With 7.7% the treatment course with Batrafen creme was repeated, and with another 7.7% was included oral antimycotic (itraconazol) . After treatment was performed microbiological check-up that confirmed the positive result of treatment . The authors' conclusion is that Batrafen creme is an effective device suitable for application on patients, CO's users.

Am J Chin Med, 1997, 25(2), 181 - 4
Antifungal activity of plant extracts against Candida albicans; Perez C et al.; In previous papers, we reported the antimicrobial activity of plants used in Argentine folk medicine against different micro-organisms . The present study deals with the screening of 11 of these plants against the opportunistic pathogen fungus Candida albicans . Aqueous extracts 6% p/v (6 mg dry plant per 100 ml of water) were checked against fungus cultures by the agar-well diffusion method . Five extracts showed antifungal activity.

Acta Haematol, 1997, 98(2), 65 - 71
Proliferative responses to PHA, anti-CD3 and antigens in patients with lymphoproliferative disease of granular lymphocytes . Mannoproteins of Candida albicans induce proliferation and cytotoxicity; Giovannetti A et al.; In this paper we investigated the lymphoproliferative and cytotoxic responses of peripheral blood mononuclear cells (PBMC) of patients affected with lymphoproliferative disease of granular lymphocytes of T cell origin to mitogens and antigens . Most patients with lymphoproliferative disease of granular lymphocytes (LDGL) showed a severely impaired PBMC proliferative capacity in response to phytohemagglutinin, anti-CD3 and tetanus toxoid, an impairment that, in the case of anti-CD3 response, appears to be related to a defect of phosphorylation in response to triggering . In contrast, at least 50% of the patients had normal proliferation in response to a mannoprotein fraction purified from Candida albicans . These data suggest that some CD3+ GL from LDGL patients, that usually respond poorly to proliferative stimuli in vitro, can be triggered to perform these functions by candidal antigens.

Int J Clin Lab Res, 1997, 27(2), 118 - 22
Benzydamine inhibits the release of tumor necrosis factor-alpha and monocyte chemotactic protein-1 by Candida albicans-stimulated human peripheral blood cells; Sironi M et al.; Benzydamine is a non-steroidal antiinflammatory drug, devoid of activity on arachidonic acid metabolism, which is extensively used as a topical drug in inflammatory conditions, particularly for the treatment of bacterial vaginosis and Candida albicans-sustained vaginitis . In the present study the effects of benzydamine on the production of several inflammatory cytokines were examined in cultures of Candida albicans-stimulated human mononuclear cells . Benzydamine (6.25-50 microM) inhibited Candida-induced tumor necrosis factor-alpha and, to a lesser extent, interleukin-1 beta production, whereas it did not affect interleukin-6 release . Benzydamine also blocked monocyte chemotactic protein-1 secretion, but it did not affect interleukin-8 production . Unlike benzydamine, ibuprofen and naproxen, two non-steroidal antiinflammatory drugs also used topically, were unable to suppress inflammatory lymphokine production from Candida-activated mononuclear cells . These data suggest that benzydamine may be effective in local Candida infections at least in part by suppressing inflammatory cytokine and monokine production in the vaginal mucosa and consequently decreasing their levels in vaginal secretions.

Scand Cardiovasc J, 1997, 31(3), 165 - 7
Post-traumatic costochondritis caused by Candida albicans . Aetiology, diagnosis and treatment; Heckenkamp J et al.; Candida costochondritis is a rare disease of complex aetiology . Pathogenetic factors range from postoperative and posttraumatic complications to haematogenous dissemination in intravenous drug addicts . In addition to clinical examination, possible diagnostic procedures include scintiscan and magnetic resonance imaging . The treatment of choice is extensive debridement and resection of the structures affected by the inflammatory process . The long-term prognosis is good.

Mycoses, 1997 Jan-Feb, 40(1-2), 47 - 54
Retrospective analysis of yeast colonization and infections in paediatric bone marrow transplant recipients; Hoppe JE et al.; Sixty-four paediatric patients who underwent allogeneic (n = 35), autologous (n = 28) or syngeneic (n = 1) bone marrow transplantation (BMT) between 1992 and 1994 were evaluated retrospectively . As antifungal prophylaxis, all patients received amphotericin B tablets and 62 of 64 (96.9%) received oral fluconazole . Weekly surveillance cultures revealed fungal colonization in 35 patients (54.7%) . Six patients (9.4%) were colonized before BMT only, 17 (26.6%) after BMT only and 12 (18.8%) both before and after BMT . Candida albicans was the most frequently isolated fungus {21 of 46 fungal isolates (45.7%)}, followed by C . glabrata {14 isolates (30.4%)} . Non-albicans species of Candida were most frequently isolated after BMT from the faeces, often in high numbers . Autologous marrow recipients had a higher fungal colonization rate both before and after BMT than allogeneic marrow recipients . One patient suffered from invasive pulmonary aspergillosis after BMT . No fungaemias or deep-seated yeast infections were observed . Six of the seven patients who had to be treated with intravenous amphotericin B because of antibiotic-refractory fever had undergone autologous BMT . Multivariate analysis of various parameters showed only pre-BMT yeast colonization to be independently associated with post-BMT colonization . Thus, systemic mycoses occurred only rarely in this study population; however yeast colonization after BMT (especially with non-albicans species) was a frequent event in spite of double prophylaxis with oral amphotericin B and fluconazole.

Mycoses, 1997 Jan-Feb, 40(1-2), 41 - 6
Activated CD8+ T cells are involved in elimination of Candida albicans from the livers of mice; Kretschmar M et al.; We have investigated the course of infection of Balb/c mice with Candida albicans . After intravenous infection of mice, the yeasts were evenly distributed in the liver sections . There was a 90% reduction in the yeasts found within 7 days after infection . This reduction was accompanied by an increase in the number of phagocytic cells (Mac-1+, Gr-1+) in cellular infiltrates around the yeast cells . Furthermore, both CD4+ and CD8+ T lymphocytes were found in these pseudogranulomata, with the CD8+ cells in the periphery . Peak amounts of the cell types investigated were found at days 5 and 6 post infection, with an increased expression of receptors for interleukin 2 (IL-2R) on the surface of CD8+ cells . At day 14 of infection, the same levels as those in control mice were reached . At this time, the yeasts were virtually eliminated from the liver . By using simultaneous detection of cell surface markers and yeast cells in immunohistology, these data demonstrate the close association of fungal cells, phagocytes, TH and TC cells in pseudogranulomata during elimination of Candida albicans from the livers of mice.

Mycoses, 1997 Jan-Feb, 40(1-2), 33 - 9
Adhesion of Candida albicans to epithelial cells--effect of nikkomycin; Segal E et al.; This study investigated the effect of the chitin synthetase inhibitors, the nikkomycins (NZ and NZ+NX), on Candida albicans adhesion to buccal epithelial cells (BECs) in vitro . The effect was expressed in reduced chitin synthetase activity and chitin content of fungal cells . In vitro adhesion assays to BECs of Candida exposed to NZ and NZ+NX revealed reduced adhesion values . Light, scanning and transmission electron microscopy (SEM, TEM) of NZ-treated and untreated micro-organisms showed changed fungal morphology and reduced adherence of the treated yeasts . Scanning electron microscopy of NZ-treated C . albicans labelled with gold-conjugated wheatgerm agglutinin (WGA) revealed less labelling than in the untreated organisms . A close contact between the fungus and the epithelial cell at a site with intense WGA-gold labelling was noted in TEM experiments . The data point to the involvement of chitin in the adhesion of C . albicans to epithelial cells.

Scand J Infect Dis, 1997, 29(3), 301 - 4
Fungemia in cancer patients undergoing chemotherapy versus surgery: risk factors, etiology and outcome; Kralovicova K et al.; 26 patients with fungemia and cancer treated with chemotherapy (group A) were compared to 25 patients with fungemia and cancer treated with surgery (group B), to assess differences in etiology, risk factors and outcome . Candida albicans was responsible for 42% of fungemias in group A, and for 92% of fungemias in group B (p < 0.005) . Breakthrough fungemia occurring during antifungal prophylaxis appeared in 46.6% of group A vs 12% of group B (p < 0.02) . There was significant difference in outcome between the groups: 20% of patients after surgery vs 7.7% of those after chemotherapy died from fungemia (p < 0.04) . Most common risk factors recorded in both groups were catheter insertion and previous therapy with broad spectrum antibiotics.

Microbios, 1997, 89(358), 23 - 8
Incubation temperatures affect adherence to plastic of Candida albicans by changing the cellular surface hydrophobicity; Blanco MT et al.; The influence of cellular surface hydrophobicity on the adherence capacity to plastic of Candida albicans was investigated at two culture temperatures (37 and 22 degrees C) . The majority of the 42 strains studied were hydrophobic at 22 degrees C and hydrophilic at 36 degrees C . The hydrophobic cells showed a consistent adherence capacity which was absent from the hydrophilic strains . The culture temperatures affect adherence to plastic of C . albicans by changing the cellular surface hydrophobicity.

Zh Mikrobiol Epidemiol Immunobiol, 1997 Jan-Feb, (1), 27 - 30
{The possibilities for using combined methods in typing Candida albicans for establishing epidemiological relationships}; Degteva GK et al.; An outbreak of candidiasis caused by C . albicans was registered in a neonatal intensive care unit of one of the hospital of St.-Petersburg at the period of August 12-October 8, 1992 . C . albicans were isolated from 21 out of 87 patients . To achieve their more detailed deciphering and to establish epidemiological relationships, the intraspecific typing of C . albicans was carried out by DNA typing and by the electrophoretic spectrum of exoproduct proteins . The analysis of the results of this typing revealed that strains classified with the same biotype belonged to different electrophoretic groups and, vice versa, strains of the same electrophoretic group could belong to different biotypes . The analysis of the data obtained by taking into account the results of typing by the two methods, the time of the patients stay in the unit, the number of the maternity clinic from which an individual patient arrived, the number of the reanimation place and the number of the apparatus for artificial lung ventilation made it possible to establish epidemiological relationships between the patients in all cases.

Microbiol Immunol, 1997, 41(5), 395 - 402
Ultrastructural alterations of Candida albicans induced by a new imidazole antimycotic omoconazole nitrate; Nishiyama Y et al.; The antifungal effects of an imidazole-antimycotic omoconazole nitrate (OMZ) on the morphology and ultrastructure of Candida albicans yeast cells were studied using scanning and transmission electron microscopy . The treatment of growing Candida cultures with fungistatic doses (0.4 to 4 micrograms/ml) of OMZ produced the formation of a chain or cluster of cells . Thickening of the cell wall and accumulation of electrondense vesicles in the wall were clearly observed . Development of Golgi-like complex membranous structures in the cytoplasm was the most prominent finding . The cytological alteration induced by exposure to a higher concentration (40 micrograms/ml) of the drug was characterized by disruption of the intracytoplasmic organelles . Our results confirm the strong antifungal activity of OMZ against fungal cells.

Microbiol Immunol, 1997, 41(5), 377 - 86
An ATP bioluminescence assay applicable to rapid fluconazole susceptibility testing of dermatophytes; Yoshida T et al.; An ATP bioluminescence assay as a rapid reference method for fluconazole (FLCZ) susceptibility testing of dermatophytes, as well as yeasts, was developed and evaluated by comparing it with viability, turbidity and fungal protein content-based conventional methods . FLCZ susceptibility results obtained with strains of Candida albicans and dermatophytes by the bioluminescence method in high-resolution medium were well correlated with those obtained by conventional methods currently used in clinical microbiology laboratories or reported previously, including a broth dilution method by the National Committee for Clinical Laboratory Standards (NCCLS) . Thus, ATP bioluminescence assay can be used to monitor fungal growth in liquid culture media . The procedure has considerable potential for the rapid testing of FLCZ susceptibility of dermatophytes and other fungi.

Urol Int, 1997, 58(3), 131 - 6
Experimental study of ascending Candida albicans pyelonephritis focusing on the hyphal form and oxidant injury; Nishikawa T et al.; To investigate the pathogenicity of the hyphal form of Candida in pyelonephritis a Candida albicans strain, assuming only the yeast form but not the hyphal form when induced by ultraviolet mutagenesis, and a revertant strain from this mutant strain showing bimorphism were compared in a rat experimental model with regard to the incidence of ascending Candida pyelonephritis and the grade of inflammation . To increase the frequency of pyelonephritis unilateral incomplete ureteral stenosis was created . The revertant strain assuming the hyphal form showed a significantly (p < 0.01) higher frequency of pyelonephritis as compared with the mutant strain not assuming this form, and the grade of inflammation was also higher in the revertant strain group . Also, higher renal tissue and serum levels of both lipid peroxide and superoxide dismutase, which are related to marked renal oxidant injury, tended to be correlated with the degree of neutrophil infiltration in the acute phase . These findings suggest that the hyphal form plays an important role in the development of C . albicans pyelonephritis and also that the oxygen radicals from neutrophils appearing at the sites of inflammation play a major part in the further extension of inflammatory lesions.

Dermatology, 1997, 194(3), 293 - 6
Immunohistochemistry with monoclonal antibody against Candida albicans mannan antigen demonstrates cutaneous Candida granulomas as evidence of Candida sepsis in an immunosuppressed host; Breier F et al.; We report the occurrence of invasive Candida albicans infection with disseminated cutaneous Candida granulomas in a patient with aplastic anaemia after viral hepatitis . Fungal elements in a skin biopsy specimen were detected by PAS stain and identified as Candida sp . by immunohistochemistry directed against the C . albicans mannan surface antigen . Based on rapid diagnosis of Candida granuloma and by Candida-positive cultures of blood and swabs, systemic treatment with liposomal amphotericin B led to survival of the patient.

Ann Pharm Fr, 1997, 55(2), 69 - 72
{Antimicrobial activity of new pyridazine derivatives}; Ungureanu M et al.; Continuing the investigations concerning synthesis-structure-biological activity in the pyridazine series, the results of the antimicrobial and antifungal tests of some new pyridazinium compounds are presented . The test was performed using the diffusimetric method with rustles steel cylinders based on the diffusion of the tested substances on the gelose surface . The comparative analysis of the obtained data leads to the following conclusions concerning the relation between structure and activity in the pyridazine series: 1 . The cis-isomers are always more active comparatively with similar trans- isomers: 2 . In the pyrrolopyridazine series we remark that the saturated or partial saturated compounds have always a stronger activity as the related aromatics derivatives . We also noticed that the saturated, partial saturated and aromatic compounds have different selectivity . Thus, the saturated are more active on the Pseudomonas aeruginosa and Candida albicans, the partial saturated are more active on the Staphylococcus aureus and Bacillus subtilis while the aromatic compounds are active on the Bacillus subtilis . 3 . The influence of the Y-substitutes of para position of benzoylic radical and the substitutes of pyrrolo ring is not decisive towards activity, these affecting especially the selectivity.

Biopolymers, 1997, 43(1), 43 - 71
Selective peptidic and peptidomimetic inhibitors of Candida albicans myristoylCoA: protein N-myristoyltransferase: a new approach to antifungal therapy; Sikorski JA et al.; MyristoylCoA: protein N-myristoyltransferase (NMT) catalyzes the cotranslational covalent attachment of a rare cellular fatty acid, myristate, to the N-terminal Gly residue of a variety of eukaryotic proteins . The myristoyl moiety is often essential for expression of the biological functions for these proteins . Attachment of C14:0 alone provides barely enough hydrophobicity to allow stable association with membranes . The partitioning of N-myrisotylproteins is therefore often modulated by "switches" that function through additional covalent or noncovalent modifications . Candida albicans, the principal cause of systemic fungal infection in immunocompromised humans, contains a single NMT gene that is essential for its viability . The functional properties of the acylCoA binding site of human and C . albicans NMT are very similar . However, there are distinct differences in their peptide binding sites . An ADP ribosylation factor (Arf) is included among the few cellular protein substrates of the fungal enzyme . Alanine scanning mutagenesis of an octapeptide derived from an N-terminal Arf sequence (GLYASKLS-NH2) disclosed that Gly1, Ser5, and Lys6 play predominant roles in binding . ALYASKLS-NH2 is an inhibitor competitive for peptide {Ki(app) = 15.3 +/- 6.4 microM} and noncompetitive for myristoylCoA . Remarkably, replacement of the N-terminal tetrapeptide with an 11-aminoundecanoyl group results in a competitive inhibitor (11-aminoundecanoyl-SKLS-NH2) that is approximately 40-fold more potent {Ki(app) = 0.40 +/- 0.03 microM} than the starting octapeptide . Removal of Leu-Ser from the C-terminus generates a competitive dipeptide inhibitor (11-aminoundecanoyl-SK-NH2) with a Ki(app) of 11.7 +/- 0.4 microM, equivalent to that of the starting octapeptide . A derivative dipeptide inhibitor containing a C-terminal N-cyclohexylethyl lysinamide moiety has the advantage of being more potent (IC50 = 0.11 +/- 0.03 microM) and resistant to digestion by cellular carboxypeptidases . Rigidifying the flexible aminoundecanoyl chain results in very potent general NMT inhibitors (IC50 = 40-50 nM) . Substituting a 2-methylimidazole for the N-terminal amine and adding a benzylic alpha-methyl group with R stereochemistry to the rigidifying element produces even more potent inhibitors (IC50 = 20-50 nM) that are up to 500-fold selective for the fungal compared to human enzyme . A related less potent member of this series of compounds is fungistatic . Its growth inhibitory effects are associated with a reduction in cellular protein N-myristoylation, judged using cellular Arf as a reporter . These studies establish that NMT is a new antifungal target.

J Drug Target, 1997, 4(5), 311 - 9
Release of amphotericin B from delivery systems and its action against fungal and mammalian cells; Legrand P et al.; Spectroscopic studies of four amphotericin B (AmB) lipid preparations--small negatively charged unilamellar vesicles "AmBisome", positively charged oligolamellar liposomes "Ampholiposomes", AmB Lipid Complex "L-AmpB33"--and AmB association with gamma cyclodextrin demonstrated that the composition of drug delivery system directly influences AmB organization . The aggregation state and release in external medium of AmB was monitored by circular dichroism and UV-visible absorption . AmB short-term activity against Candida albicans (K+ leakage) was found to be correlated with the amount of free AmB released from lipid preparations . These data seem to indicate that lipid composition influences anti-Candida albicans activity, by modulation of AmB binding to lipids.

Dermatology, 1997, 194 Suppl 1, 27 - 31
Effect of Lamisil and azole antifungals in experimental nail infection; Richardson MD; Onychomycosis is primarily caused by dermatophyte fungi but occasionally by yeasts and non-dermatophytic moulds . The aim of this study was to develop an in vitro model of nail invasion by dermatophytes, yeasts and non-dermatophytic moulds, and to provide an alternative system for studying the activity of different classes of antifungal drugs against fungi associated with onychomycosis . In the absence of extraneous nutrients, Trichophyton mentagrophytes was seen in electron microscopy to degrade completely healthy nail plate . Candida albicans germinated on nail fragments, but invasion of the nail plate was not seen . The mould Fusarium formed long channels through the matrix of the nail plate . Aspergillus versicolor appeared to penetrate the outer and intermediate surface of the nail plate only . Acremonium sp . and Scopulariopsis brevicaulis did not invade nail in this model . Exposure of nail fragments to terbinafine (0.25 mg/l for 3 h) inhibited invasion by T . mentagrophytes, C . albicans and the non-dermatophytic moulds . Itraconazole (0.25 mg/l for 3 h) prevented nail plate invasion by T . mentagrophytes, A . versicolor and Fusarium but did not totally inhibit the surface growth of Acremonium or S . brevicaulis . C . albicans grew in the presence of itraconazole . The results indicate that terbinafine is readily absorbed by the nail and that the drug is bio-available in nail keratin . A short exposure of nail to low concentrations of terbinafine acted as a barrier against fungal invasion . Itraconazole appeared to be effective against Trichophyton and some non-dermatophytic moulds.

Peptides, 1997, 18(3), 415 - 22
Inhibition of human neutrophil functions by sulfated and nonsulfated cholecystokinin octapeptides; Carrasco M et al.; The effects of CCK-8s and desulfated CCK-8 at concentrations ranging from 10(-14) to 10(-6) M were studied in vitro on several functions of human peripheral neutrophils: adherence to substrate, mobility (spontaneous and directed by a chemical gradient or chemotaxis), ingestion of inert particles (latex beads) or cells (Candida albicans), and production of superoxide anion measured by the nitroblue tetrazolium reduction test . The effect of CCK-8s on intracellular levels of cAMP was investigated as well as the implication of calcium in the action of CCK-8s on phagocytic function using stimulants and inhibitors of both intracellular and extracellular calcium channels . The two peptides, at concentrations from 10(-12) to 10(-8) M, inhibited significantly both mobility and ingestion capacities and increased adherence to substrate . A dose-response relationship was observed with a maximum inhibition of neutrophil functions at 10(-10) M, CCK-8s and desulfated CCK-8 induced in these cells a significant, but transient, increase of cAMP levels at 60 s . Moreover, CCK-8s was found to inhibit completely the stimulation of latex bead phagocytosis in neutrophils produced by the calcium ionophore A23187 . These results suggest that CCK-8 is a negative modulator of several neutrophil functions and that the inhibition of these activities could be carried out through an increase of the intracellular cAMP levels and a decrease of the extracellular calcium input.

ASAIO J, 1997 Jan-Feb, 43(1), 23 - 30
Nonaldehyde sterilization of biologic tissue for use in implantable medical devices; Moore MA et al.; Biologic tissue stabilized by dye-mediated photooxidation has found application in implantable devices . The desire to avoid aldehydes in the processing of photooxidized tissues led to the development of a nonaldehyde, iodine based sterilant . The interaction of tissue with iodine was indicated by a change in tissue shrinkage temperature, dependent upon solution and incubation parameters . The amino acid tyrosine also was altered, presumably because of aromatic ring iodination . Transmission electron microscopic study indicated no change in the quarter staggered array structure of collagen under controlled iodine treatment conditions . The D10 values for iodine kill of several organisms, in the absence of tissue, were determined in 0.1% iodine (pH 6.5) at 37 degrees C for Bacillus subtilis (12 min, Aspergillus niger, Escherichia coli, Candida albicans, Staphylococcus aureus, and Pseudomonas aeruginosa (all < 1 min) . In a separate experiment, samples of 0.1% iodine (pH 6.5) containing photooxidized pericardial tissue were inoculated with 1.6 X 10(7) Bacillus subtilis, 4.6 X 10(6) Pseudomonas aeruginosa, or 7.2 X 10(6) Staphylococcus aureus and incubated at 37 degrees C . No survivors were detected on the tissue samples after exposure of 48 hr . Photooxidized pericardial tissue samples inoculated with either 3.2 X 10(5) porcine parvovirus or 1 X 10(9) infectious bovine rhinotracheitis were exposed to 0.1% iodine (pH 6.5) at 36 degrees C for 12 h . No viral particles were detected after exposure, yielding minimum viral log reduction factors of 3.0 and 6.5, respectively . The results presented indicate the potential for the nonaldehyde, iodine based solution to sterilize implantable devices containing biologic tissue.

Drugs Exp Clin Res, 1997, 23(1), 33 - 8
Effects of nedocromil sodium on the oxidative burst of polymorphonuclear leukocytes: comparison with salbutamol; Braga PC et al.; The effects of nedocromil sodium and of salbutamol on the generation of oxygen-derived free radicals in human polymorphonuclear leukocytes (PMNs) were compared in vitro by the luminol-amplified-chemiluminescence (LACL) assay induced by both particulate (Candida albicans) and soluble formyl-methionyl-leucyl-phenylalanine (fMLP) stimulants . Inhibitory dose-effect linear regressions were observed from 10(-3) to 10(-8) M for nedocromil and salbutamol after a 3' period of incubation with either C . albicans or fMLP . There was a linear regression with nedocromil sodium after 30' incubation, but desensitization was observed with salbutamol after this longer period of incubation . The generation of oxygen-derived free radicals was significantly greater for asthmatic patients than for normal subjects; therefore antiasthmatic drugs with this inhibitory activity could be an extra pharmacological benefit in the treatment of asthmatic patients.

Boll Chim Farm, 1997 Jan, 136(1), 17 - 21
Application of triphenyltetrazolium chloride in microbial limit test in pharmaceuticals and cosmetics . Multiple- tube method for yeasts detection; Ohara MT et al.; The triphenyltetrazolium chloride (TTC) was studied as a means to detect yeasts growth in liquid medium, aiming its application to microbial limit test using Multiple-tube method, for testing pharmaceuticals or cosmetics samples with opacity . The results from most probable number (MPN) using TTC and sub-culture techniques obtained from the tests of simulated samples, spiked with Candida albicans and Sacharomyces cerevisiae, showed to be equivalents.

Eur J Clin Microbiol Infect Dis, 1997 Jan, 16(1), 16 - 20
Clearances of Candida albicans-derived alpha- and beta-linked mannose residues in sera from patients with candidiasis; Poulain D et al.; Two monoclonal antibodies (MAbs) reacting with different Candida albicans mannan oligomannosidic epitopes were used to detect antigens in human sera . The first MAb reacted with alpha-linked mannose residues shared by mannoproteins, and the second displayed reactivity against beta-1,2-linked oligomannosides shared by phospholipomannan . Two latex agglutination tests performed after dissociation of serum immune complexes were 100% specific . In a retrospective analysis of 39 sera from 20 patients with candidiasis, each test detected seven patients; in combination, they detected ten . More than 60% of positive samples were positive with only one test . These results demonstrate that the clearance of Candida albicans antigens from the blood differs according to the type of oligomannoside and glycoconjugate . Antigen detection kits with MAbs to different Candida albicans mannan epitopes could provide a logical rationale to compensate for the transitory character of mannanemia detection during candidal infections.

J Clin Lab Anal, 1997, 11(2), 104 - 9
Plasma (1-->3)-beta-D-glucan assay and immunohistochemical staining of (1-->3)-beta-D-glucan in the fungal cell walls using a novel horseshoe crab protein (T-GBP) that specifically binds to (1-->3)-beta-D-glucan; Tamura H et al.; A highly sensitive enzyme-linked immunosorbent assay specific to (1-->3)-beta-D-glucans (GBP-ELISA) has been developed using a novel (1-->3)-beta-D-glucan-binding protein (T-GBP), which was purified from the amebocyte lysate of the Japanese horseshoe crab, Tachypleus tridentatus . This method allowed quantitation of the glucans in a concentration range of 0.1-1,000 ng/ml, regardless of linear and branched structures, and was applied to determine the amounts of (1-->3)-beta-D-glucan in human and animal plasmas for diagnosis of fungemia . High levels of plasma glucan contents in clinical samples were found to be correlated closely with the severity of fungal infection . T-GBP was successfully utilized for indirect immunofluorescence staining of (1-->3)-beta-D-glucan in Candida albicans cell walls.

Intensive Care Med, 1997 Jan, 23(1), 23 - 30
Candidemia in non-neutropenic critically ill patients: analysis of prognostic factors and assessment of systemic antifungal therapy . Study Group of Fungal Infection in the ICU; Nolla-Salas J et al.; OBJECTIVE: To determine the incidence and prognosis of candidemia in non-neutropenic critically ill patients, to define mortality-related factors, and to evaluate the results of systemic antifungal therapy . DESIGN: A prospective multicenter survey in which medical and/or surgical intensive care units (ICUs) in 28 hospitals in Spain participated . PATIENTS: All critically ill patients with positive blood cultures for Candida species admitted to the participating ICUs over a 15-month period were included . INTERVENTIONS: Candidemia was defined as the presence of at least one positive blood culture containing Candida species . The follow-up period was defined as the time elapsed from the first positive blood culture for Candida species to discharge or death during hospitalization . Antifungal therapy was considered to be "early" when it was administered within 48 h of the date when the first positive blood culture was obtained and "late" when it was administered more than 48 h after the first positive blood culture . MEASUREMENTS AND MAIN RESULTS: Candidemia was diagnosed in 46 patients (mean age 59 years), with an incidence of 1 critically ill patient per 500 ICU admissions . The species most frequently isolated were Candida albicans (60%) and C . parapsilosis (17%) . Fluconazole alone was given to 27 patients, amphotericin B alone to 10, and sequential therapy to 6 . Three patients did not receive antifungal therapy . The overall mortality was 56% and the attributable mortality 21.7% . In the univariate analysis, mortality was significantly associated with a higher Acute Physiology and Chronic Health Evaluation (APACHE) II score at the onset of candidemia (p = 0.04) and with the time elapsed between the episode of candidemia and the start of antifungal therapy 48 h or more later (p < 0.02) . Patients with an APACHE II score lower than 21 at the onset of candidemia had a higher probability of survival than patients who were more seriously ill (p = 0.04) . Patients with "early" antifungal therapy (< or = 48 h between the onset of candidemia and the start of antifungal therapy) had a higher probability of survival compared with patients with late therapy (p = 0.06) . No significant differences were noted between the two groups on different antifungal therapy . CONCLUSIONS: The incidence of candidemia in ICU patients was very low . An APACHE II score > 20 at the time of candidemia was associated with a higher mortality . Further studies with a large number of patients are needed to assess the effect of early antifungal therapy on the decrease in mortality associated with candidemia and to determine the appropriate dosage of fluconazole and duration of treatment.

J Ethnopharmacol, 1997 Jan, 55(2), 151 - 9
Anticandidal activity of Santolina chamaecyparissus volatile oil; Suresh B et al.; A search for naturally occurring drugs with antifungal activity lead to Santolina oil, a volatile oil distillate of Santolina chamaecyparissus . The studies revealed that Santolina oil was effective in controlling experimental candidiasis in vitro and in vivo . It had a synergistic effect on clotrimazole in controlling Candida albicans in vitro . It significantly controlled experimental vaginal candidiasis and experimental systemic candidosis . Santolina oil was able to control the superficial cutaneous mycoses . It is recommended as a potential candidate for further studies, including clinical studies.

Comp Immunol Microbiol Infect Dis, 1997 Jan, 20(1), 21 - 7
Effect of serum from breast- or formula-fed infants on polymorphonuclear leukocyte function; Barriga Ibars C et al.; The aim of the present study was to determine the influence of serum from formula and breast-fed infants on neutrophil function (as measured by the attachment and phagocytosis of Candida albicans) as well as the chemoattractant activity of the serum . The results indicate that: (a) serum from breast-fed infants induces a greater chemoattractant activity in neutrophils than serum from 3-month-old formula-fed infants; (b) the highest values of the attachment capacity were obtained after incubation of neutrophils with serum from 1-month-old breast-fed infants; and (c) serum from breast-fed infants induces a greater phagocytic capacity against C . albicans in neutrophils than serum from formula-fed infants.

Clin Diagn Lab Immunol, 1997 Jan, 4(1), 96 - 103
Reconstruction of the immune system after unrelated or partially matched T-cell-depleted bone marrow transplantation in children: functional analyses of lymphocytes and correlation with immunophenotypic recovery following transplantation; Kook H et al.; Reconstitution of the immune system following T-cell-depleted bone marrow transplantation (BMT) in children has yet to be fully elucidated . Thus, we prospectively studied the recovery of immune function in 64 children who underwent T-lymphocyte-depleted marrow transplants using either matched family member donors or matched unrelated donors . We measured in vitro posttransplantation proliferative responses to phytohemagglutinin (PHA), concanavalin A, pokeweed mitogen, and Candida albicans antigen and assessed unidirectional allogeneic mixed-lymphocyte culture (MLC) responses at various times . A total of 129 healthy individuals served as normal controls for these assays . Responses to T-cell mitogens normalized within 12 months posttransplantation, while MLC responses normalized by 9 months . The presence of graft-versus-host disease (grade II or greater) and cytomegalovirus infection was associated with delays in immune function recovery . Importantly, immune function recovery correlated temporally with a rise in peripheral lymphocyte count . In contrast, the CD4/CD8 ratio was not predictive of immune recovery . Knowledge of immune function recovery may guide clinicians in devising strategies to minimize the risk of infection post-BMT.

Am J Respir Crit Care Med, 1997 Jan, 155(1), 87 - 93
Allergen exposure decreases glucocorticoid receptor binding affinity and steroid responsiveness in atopic asthmatics; Nimmagadda SR et al.; Allergen exposure can confound the management of asthma . To understand the potential mechanisms by which allergens increase the steroid requirements in atopic asthmatics, we examined the effects of allergens on glucocorticoid receptor (GCR) binding affinity and glucocorticoid (GC) responsiveness of peripheral blood mononuclear cells (PBMC) from atopic asthmatics . A significant reduction (p < 0.001) in the GCR binding affinity (Kd) was observed in ragweed-allergic asthmatics during ragweed pollen season compared with PBMC obtained before and after ragweed season . In vitro effects of allergen on PBMC GCR Kd were also examined by incubating PBMC from atopic asthmatics with allergen (ragweed and cat) versus Candida albicans . GCR binding affinity was significantly reduced after incubation with ragweed (p < 0.001) or cat allergen (p < 0.001) compared with baseline or C . albicans stimulation . This effect was limited to atopic asthmatics in that in vitro cat allergen incubation for 48 h failed to significantly alter GCR binding affinity in nonasthmatic, atopic individuals . These allergen-induced reductions in GCR binding affinity also rendered the PBMC less sensitive to the inhibitory effects of hydrocortisone and dexamethasone on allergen-induced proliferation (p < 0.01) . To test the hypothesis that allergen-induced alterations in GCR binding affinity were cytokine-induced, we examined the effects of interleukin-2 (IL-2)and IL-4 neutralization using anticytokine antibodies . Addition of both anti-lL-2 and anti-lL-4 antibodies resulted in a significant (p < 0.001) inhibition of allergen-induced alterations in GCR binding affinity . Furthermore incubation with cat allergen induced significantly higher concentrations of IL-2 (p = 0.03) and IL-4 (p = 0.02) by PBMC from atopic as compared with nonatopic subjects . Our current observations suggest that allergen exposure may contribute to poor asthma control by reducing GCR binding affinity in mononuclear cells . This appears to be mediated through IL-2 and IL-4 . These findings may have important implications for novel approaches to the treatment of poorly controlled asthma.

Curr Genet, 1997 Jan, 31(1), 1 - 6
Characterization of the Saccharomyces cerevisiae FCY1 gene encoding cytosine deaminase and its homologue FCA1 of Candida albicans; Erbs P et al.; By functional complementation of a fcy1 null mutant of Saccharomyces cerevisiae, we have cloned and characterized the FCY1 gene, encoding cytosine deaminase in Saccharomyces cerevisiae, and its homologue FCA1, encoding cytosine deaminase in Candida albicans . Disruption of FCY1 resulted in high resistance to 5-fluorocytosine (10(-2) M) and in total loss of cytosine deaminase activity . By contrast the transformation by FCY1 or FCA1 of the haploid FCY1-disrupted host strain restored sensitivity to 5-fluorocytosine and allowed growth on cytosine, as a source of pyrimidine, or ammonium . FCA1 as opposed to FCY1 contains an intron . FCA1 and FCY1 encode respectively 150- and 158- residue proteins of 60% identity . Both Fcy1p and Fca1p share common motifs with cytidine and CMP deaminases, but homology with cytosine deaminase of E . coli could not be detected.

FEMS Microbiol Lett, 1997 Jan 1, 146(1), 65 - 71
Thermotolerance and trehalose accumulation induced by heat shock in yeast cells of Candida albicans; Arguelles JC; Candida albicans yeast cells growing exponentially on glucose are extremely sensitive to severe heat shock treatments (52.5 degrees C for 5 min) . When these cultures were subjected to a mild temperature preincubation (42 degrees C), they became thermotolerant and displayed higher resistance to further heat stress . The intracellular content of trehalose was very low in exponential cells, but underwent a marked increase upon non-lethal heat exposure . The accumulation of trehalose is likely due to heat-induced activation of the trehalose-6-phosphate synthase complex, whereas the external trehalase remained practically unmodified . After a temperature reversion shift (from 42 degrees C to 28 degrees C), the pool of trehalose was rapidly mobilized without any concomitant change in trehalase activity . These results support an important role of trehalose in the mechanism of acquired thermotolerance in C . albicans and seem to exclude the external trehalase as a key enzyme in this process.

Clin Infect Dis, 1997 Jan, 24(1), 28 - 34
Infection due to fluconazole-resistant Candida in patients with AIDS: prevalence and microbiology; Maenza JR et al.; A cross-sectional study was conducted to assess the prevalence and microbiology of oral infection due to fluconazole-resistant Candida in patients with AIDS . Oral swab specimens for fungal cultures were obtained from 100 consecutive outpatients with CD4 lymphocyte counts of < 200/mm3 . At least one fungal organism demonstrating in vitro resistance to fluconazole (minimum inhibitory concentration, > or = 8 micrograms/mL) was isolated from 26 (41%) of 64 patients for whom cultures were positive . When fluconazole-resistant C . albicans was isolated, in vitro resistance correlated with clinical thrush . None of 10 patients from whom only non-albicans species of Candida were isolated had active thrush . The patients from whom fluconazole-resistant Candida albicans was isolated had lower CD4 cell counts (median, 9/mm3), a greater number of treated episodes of thrush (median, 4.5), and a greater median duration of prior fluconazole treatment (231 days) than did patients from whom fluconazole-susceptible C . albicans was isolated (median CD4 cell count, 58/mm3 {P = .004}; median number of treated episodes of thrush, 2.0 {P = .001}; and median duration of prior fluconazole treatment, 10 days {P = .01}; respectively) . In a multivariate analysis, the number of episodes and duration of fluconazole therapy were independent predictors of resistance.

Crit Care Med, 1997 Jan, 25(1), 111 - 20
The yeast to hyphal transition following hematogenous candidiasis induces shock and organ injury independent of circulating tumor necrosis factor-alpha; Matuschak GM et al.; OBJECTIVES: Dimorphic Candida albicans spp . increasingly cause lethal septic shock and disseminated infection in the critically ill . Following candidemia, production of specific fungal exotoxins coincident with the yeast to hyphal phenotypic transition is believed to be important in the pathogenesis of Candida septic shock . However, overexpression of the pleiotropic cytokine tumor necrosis factor (TNF)-alpha by the host following hyphal germination is also thought to be a mechanism of Candida-related cardiopulmonary dysfunction, as well as of bacteremic shock . In this study, we hypothesized that increases in circulating TNF-alpha coinciding with the yeast to hyphal transition modulate the onset and progression of shock with multiple organ injury early after hematogenous candidiasis . DESIGN: Prospective, controlled laboratory animal study . SETTING: University hospital animal research facility . SUBJECTS: Pathogen-free, male Sprague-Dawley rats (n = 26) . INTERVENTIONS: Conscious, antibiotic-treated animals with chronic indwelling carotid arterial and jugular venous catheters were intravenously infected with 10(9) viable blastoconidia of the C . albicans clinical pathogen, CA-MEN (n = 10), over 30 mins and ending at t = 0 hr, compared with an equivalent inoculum of its viable agerminative mutant, CA-MM2002 n = 11), or an intravenous infusion of 0.9% sodium chloride (n = 5) . MEASUREMENTS AND MAIN RESULTS: Mean arterial pressure (MAP), pulse rate, respiratory frequency, rectal temperature, acid-base status, quantitative blood cultures, circulating alanine aminotransferase (ALT), and bioactive TNF-alpha were serially measured in all three groups over 24 hrs or until death . Organ cultures, wet/dry weight ratios, and histopathologic changes in the lungs, heart, liver, and kidneys were determined in Candida-infected and 0.9% sodium chloride (normal saline)-infused subgroups at 6 and 24 hrs . Animals hematogenously infected with the C . albicans clinical isolate developed lethal nonendotoxemic shock in < or = 6 hrs (MAP 49 +/- 7 mm Hg {SEM}; p < .05 vs . t = 0 hr), and at death (7.0 +/- 0.3 hrs) were acidotic, hypocapnic, and hypothermic (rectal temperature 33.2 +/- 0.7 degrees C) . Despite similar peak concentrations of circulating fungal colony-forming units (cfu) and kinetics of vascular clearance in both Candida-infected groups, survival and MAP in rats challenged with the agerminative C . albicans mutant were unchanged for > 8 hrs, as were pH, Pco2, and rectal temperature . No germination of the agerminative fungal strain occurred in vivo over 6 hrs . Serum TNF was nearly undetectable at t = 0 hr in all three groups . Although shock developed soon after fungemia with the C . albicans clinical isolate, TNF-alpha concentrations did not increase above normal saline values in either candidemic group at t = 1.5, 4.5, or 6 hrs (17 +/- 7 vs . 14 +/- 1 U/mL in the parent C . albicans organism vs . its agerminative mutant at t = 6 hrs) . Greater numbers of agerminative C . albicans than its dimorphic parent strain were recovered from the lungs (5.41 +/- 1.0 vs . 2.02 +/- 0.38 x 10(7) cfu/g, respectively; p < .05) and kidneys (p < .01) . By 24 hrs, modest germination of the mutant Candida strain was observed in the tissues . However, lung wet/dry ratios, intrapulmonary hyphal proliferation, and alveolar hemorrhage were all greater after infection with the parent fungal isolate . Likewise, myocardial necrosis and hepatic glycogen depletion with vacuolization were more severe after infection with the C . albicans clinical isolate vs . candidemia with its agerminative mutant, although serum ALT values did not differ between these groups . CONCLUSIONS: Lethal C . albicans sepsis with lung injury and multiple organ damage are temporally associated with the in vivo yeast to hyphal transition in this model . However, this candidemic septic shock syndrome is modulated by circulating fungal virulence factors or host mediators other than TNF-alpha, a cytokine considered essen

Free Radic Biol Med, 1997, 22(3), 561 - 5
Scavenging of reactive oxygen species by a glycolipid fraction of Mycobacterium avium serovar 2; Scherer TA et al.; Previous experiments indicated that MIF-A3, a peptidoglycolipid extracted from Mycobacterium avium serovar 2 (Mycobacterium paratuberculosis 18), inhibits the killing of Candida albicans by activated bovine peripheral blood-derived macrophages and murine thioglycollate-elicited peritoneal macrophages in vitro . Subsequent in vitro data from our laboratory indicated that this reduction in killing may be related to the ability of MIF-A3 to scavenge reactive oxygen species (ROS) . In this study we examined this hypothesis directly by determining if MIF-A3 reduced exogenous H2O2-induced candidacidal activity . When Candida albicans was incubated with H2O2 (4 mM) alone, colony-forming units/ml x 10(4) (CFU/ml) were 0.4 +/- 0.1 (mean +/- SE, n = 4) as compared to 11.3 +/- 2.0 CFU/ml in control (untreated) cultures (p < .05) . The addition of catalase at concentrations > or = 6.8 U/ml, completely blocked the fungicidal effect of H2O2 . However, reducing the amount of catalase from 6.8 U/ml to 3.4 U/ml resulted in a loss of scavenging activity, which was associated with a 50% increase in H2O2-mediated killing . Substituting MIF-A3 (400 micrograms/ml) for catalase, also reduced H2O2-induced fungicidal activity . In the absence of MIF-A3, H2O2 reduced Candida albicans to less than 10(3) CFU/ml . However, in the presence of MIF-A3 the CFU/ml of Candida albicans increased 7.5-fold . Based on concentration-response curves of H2O2 inhibition vs . increasing amounts of catalase we determined that the relative inhibitory capacity of the MIF-A3 (400 micrograms/ml) was approximately 1.0 U/ml "catalase equivalents." These findings provide direct evidence that MIF-A3 can scavenge H2O2, and reduce H2O2-induced killing of Candida albicans.

Antimicrob Agents Chemother, 1997 Jan, 41(1), 196 - 9
Isolation and characterization of fluconazole- and amphotericin B-resistant Candida albicans from blood of two patients with leukemia; Nolte FS et al.; Infections with fluconazole-resistant Candida albicans isolate have rarely been described in clinical settings other than oropharyngeal candidiasis in patients with late-stage AIDS . We report on two patients with leukemia who developed fungemia caused by fluconazole-resistant C . albicans after receiving fluconazole prophylaxis (400 mg/day) and empiric amphotericin B therapy (0.5 mg/kg of body weight per day) . The fluconazole MICs for the isolates were > or = 64 micrograms/ml, and the isolates were resistant to other azoles and had membrane sterol changes consistent with a mutation in the delta 5,6-sterol desaturase gene . The lack of ergosterol in the cytoplasmic membrane of the fluconazole-resistant strains also imparted resistance to amphotericin B . Both patients were successfully treated with high-dose amphotericin B (1 to 1.25 mg/kg/day) and flucytosine (150 mg/kg/day).

J Clin Microbiol, 1997 Jan, 35(1), 313 - 6
Candida and Torulopsis: a blinded evaluation of use of pseudohypha formation as basis for identification of medically important yeasts; Odds FC et al.; Seventy yeast isolates representing species in the genera Candida and Torulopsis but excluding Candida albicans were examined in three laboratories for production of pseudohyphae in Dalmau cultures . The microscopic morphology of the isolates was scrutinized by four individuals experienced in yeast identification and three inexperienced persons, all of whom were blinded as to the putative identification of the yeasts . For 49 (70%) of the 70 isolates, the seven observers recorded comparable scores for morphology, but 5 (7%) of the isolates showed extreme variation in recorded morphologies, from true hyphae formed to no pseudohyphae formed . Isolates of Candida parapsilosis and Torulopsis glabrata consistently did and did not form pseudohyphae, respectively: however, other Candida and Torulopsis spp . did not always express their expected morphologies . In 48 (19%) of 252 readings (seven observers), 36 isolates of Candida spp . were scored as forming no pseudohyphae, and in 22 (9.2%) of 238 readings, 34 isolates of Torulopsis spp . were recorded as forming true hyphae or pseudohyphae . These results show that pseudohypha formation is not a reliable characteristic for identification of yeasts at the genus level; we suggest that the merger of Torulopsis spp . into the genus Candida should be finally accepted.

J Clin Microbiol, 1997 Jan, 35(1), 5 - 10
Rapid susceptibility testing of fungi by flow cytometry using vital staining; Wenisch C et al.; A 1-h assay for antifungal susceptibility testing measuring the impairment of fungal metabolic activity was developed . Yeast viability was analyzed by flow cytometry with a novel fluorescent probe, FUN-1, which emits a red fluorescence when the yeast is metabolically active . For nine Candida albicans strains tested, this method yielded results comparable to those obtained by the standard M27 procedure for amphotericin B, flucytosine, fluconazole, and ketoconazole . Whether the flow cytometry antifungal susceptibility test results correlate with the in vivo activities of the drugs remains to determined.

FEMS Immunol Med Microbiol, 1996 Dec 31, 16(3-4), 229 - 34
Over-expression of Saccharomyces cerevisiae hsp90 enhances the virulence of this yeast in mice; Hodgetts S et al.; Saccharomyces cerevisiae, a yeast of low pathogenic potential, is a rare but well-documented cause of invasive infections in humans . The yeast Candida albicans is a much commoner cause of significant and life-threatening infections . In such infections the heat shock protein hsp90 is an immunodominant antigen associated with protective humoral immunity . In this study it was shown that over-expression of S . cerevisiae hsp90, the amino acid sequence of which shows 84% identity to C . albicans hsp90, significantly increased the virulence of a laboratory strain of S . cerevisiae in mice, both in terms of colony counts in the kidney, liver, and spleen, and in terms of mortality . This is the first direct evidence that hsp90 is a virulence factor.

Wien Klin Wochenschr, 1996 Dec 27, 108(24), 795 - 801
{Early postoperative infections after liver transplantation--pathogen spectrum and risk factors}; Gotzinger P et al.; Infections occurring during the early postoperative phase after liver transplantation result in a significant rise in morbidity and mortality . The records of 279 orthoptic transplantations performed in 248 patients were analyzed retrospectively . 55.6% of all patients suffered from one or more episodes of bacterial and/or fungal infection during their postoperative hospitalisation . The median onset of bacterial/fungal infection was on day 7 after transplantation . Enterococci (42 episodes), Pseudomonas aeruginosa (38 episodes), staphylococci (37 episodes), Escherichia coli (17 episodes) and Candida albicans (11 episodes) were the most frequently detected organisms . 74 (29.8%) patients developed viral infections . 20 patients (8.1%) showed infection with cytomegalovirus (CMV), 32 patients (12.9%) with herpes simplex virus (HSV) and 6 patients (2.4%) with varicella zoster virus (VZV) . 14 patients (5.6%) developed infection with both CMV and VZV . Triple infection with CMV, HSV and VZV occurred in one patient . Statistical analysis of potential risk factors showed a significant influence of blood volume replacement (p < 0.001) and occurrence of at least one rejection period (p < 0.02) for major bacterial/fungal infection and immunosuppression (p < 0.001), cold ischemic time (p < 0.04), occurrence of at least one rejection period (p < 0.005) and blood volume replacement (p < 0.04) for viral infection.

Gene, 1996 Dec 12, 183(1-2), 159 - 65
Isolation and molecular characterisation of the POL3 gene from Candida albicans; Nolan T et al.; In Saccharomyces cerevisiae, the CDC2 gene encodes the large subunit of DNA polymerase III, the analogue of mammalian DNA polymerase delta . We have isolated DNA fragments from a library of Candida albicans genomic DNA in the vector pRS316 that rescue temperature sensitive cdc2 mutations in S . cerevisiae . These fragments contain an ORF coding for a protein of 1038 aa with a predicted molecular mass of 118.8 kDa . The predicted protein shows homology to a number of eukaryotic DNA polymerases, with 62% identity over its length to the S . cerevisiae Cdc2 protein . It also contains a number of motifs which are characteristic of DNA polymerases in general and viral polymerases in particular, as well as the conserved motif which interacts with proliferating cell nuclear antigen . These results indicate that this gene is C . albicans POL3 . Analysis of the expression of C . albicans POL3 revealed that the transcript is present throughout the mitotic cell cycle, which contrasts with the expression of S . cerevisiae CDC2.

J Diarrhoeal Dis Res, 1996 Dec, 14(4), 286 - 8
Epithelial adherence of Candida albicans is enhanced by passage through rat small intestine; Nath G et al.; Seven Candida albicans isolates (four from patients with diarrhoea and three from healthy persons) underwent two passages through rat ileal loop (RIL) to see the effect of consecutive passages on the adherence to rat intestinal epithelium . The isolates from patients with diarrhoea showed a significant enhancement in adherence after the first passage (1.95 x 10(4) cfu/cm2 versus 3.67 x 10(4) cfu/cm2) . There was no further increase between the first passage (3.67 x 10(4) cfu/cm2) and the second one (3.61 x 10(4) cfu/cm2) . A similar pattern was observed with the three nondiarrhoeal isolates . Animal passage of this fungus probably leads to better interactions between the cell surfaces causing the enhanced adherence.

Rev Esp Fisiol, 1996 Dec, 52(4), 215 - 22
Comparative study of the effect of teicoplanin and vancomycin upon the phagocytic process of peritoneal macrophages; Barriga C et al.; The effect of Teicoplanin and Vancomycin upon the phagocytic process was compared by evaluating the different activities of the peritoneal macrophage phagocytic function from mice treated with these antibiotics . The results indicated that teicoplanin and vancomycin increased both the substrate adherence and chemotaxis of peritoneal macrophages and that neither antibiotic, at the concentrations of 10, 25, 50, 75 and 100 mg/l, had any chemoattractant capacity for peritoneal macrophages . There was an increase in the attachment of Candida albicans only in the macrophages from mice treated with vancomycin . The phagocytosis of C . albicans, Staphylococcus aureus, Escherichia coli and inert particles as well, as the nitroblue tetrazolium reduction capacity (microbicide capacity) increased in the presence of both antibiotics . The C . albicans digestion capacity increased only in the peritoneal macrophages from mice treated with teicoplanin.

Antimicrob Agents Chemother, 1996 Dec, 40(12), 2835 - 41
Multiple efflux mechanisms are involved in Candida albicans fluconazole resistance; Albertson GD et al.; Fluconazole-susceptible Candida albicans strains accumulated {3H}fluconazole at a rate of approximately 2 pmol/min per 10(9) cells . Fluconazole accumulation was not affected by the pretreatment of cells with sodium azide or with 2-deoxyglucose . The rate of fluconazole accumulation became saturated at high fluconazole concentrations and was not affected by the addition of ketoconazole, and there was no fluconazole accumulation in cells incubated at 4 degrees C . A fluconazole-resistant mutant of C . albicans SGY-243 was isolated following growth enrichment in fluconazole-containing medium . Cells of the mutant strain, designated FR2, showed a reduced rate of fluconazole accumulation compared with SGY-243 and were not resistant to other azole antifungal agents . The rates of fluconazole accumulation by C . albicans FR2 and the other azole-resistant strains, B59630, AD, and KB, were increased in the presence of sodium azide, suggesting that fluconazole resistance in these strains may be associated with an energy-dependent drug efflux . Fluconazole-resistant C . albicans strains all contained elevated amounts (2- to 17-fold) of mRNA encoding Cdr1, and an ATP-binding cassette-type transporter . In addition, C . albicans FR2 also contained increased amounts of mRNA encoding Benr, a major facilitator superfamily transporter . These results suggest that fluconazole enters C . albicans cells by facilitated diffusion and that fluconazole resistance may involve energy-dependent drug efflux associated with increased expression of Benr and/or Cdr1.

Versicherungsmedizin, 1996 Dec 1, 48(6), 215 - 7
{Fungi in feces, fungi in the intestines--therapeutic consequences?}; Rosch W; Yeast in stool specimen are due to transient or commensal growth in the GI tract . Only in immune deficient subjects candida albicans may grow invasively in squamous epithelium . In dermal or vaginal mycosis systemic therapy does not add benefit to local measures . Candida-induced diarrhea in hospitalized patients following chemotherapy stop after a few days of nystatin treatment . Candida hypersensitivity syndrome does not exist, antifungal diet does not eradicate yeast . Stool examination for candida is of no sense because a positive finding is seen in up to 80% of healthy persons.

Epidemiol Mikrobiol Imunol, 1996 Dec, 45(4), 163 - 70
{Newly emerging fungal pathogens}; Tomsikova A; Candida albicans was once considered the only important fungus species associated with human infection . Modern medical therapy and improved methods for detection and differentiation of fungi have now shown that many other species are clinically important . During the past decade, is has become apparent that species once considered only of industrial importance or harmless, are capable to attack the human host . These organisms can vary greatly.

Enferm Infecc Microbiol Clin, 1996 Dec, 14(10), 586 - 9
{Utility of Fluoroplate Candida for the rapid identification of Candida albicans}; Quindos G et al.; BACKGROUND: Candida albicans infections are frequent in immunocompromised patients and a prompt diagnosis could favor an early and proper antifungal treatment . The rapid identification of clinical yeast isolates facilitate this diagnosis . METHODS: The utility of Fluoroplate Candida ready-to-use plates for Candida albicans rapid identification was evaluated with 653 clinical isolates from 23 yeast species, including 307 C . albicans plated onto Fluoroplate Candida agar (Merck, Germany) . Rapid identification of C . albicans was based on the hydrolysis of 4-methylumbelliferyl-N-acetyl-beta-D-galactosaminide by the galactosaminidase activity of C . albicans producing white fluorescent colonies under ultraviolet light . Identification on Fluoroplate Candida was confirmed by germ tube, chlamydoconidia formation and API-ATB ID 32C assays . RESULTS: Three hundred and five of 306 isolates showing fluorescent colonies were C . albicans and one was Candida glabrata (false positive) . The rest of the isolates showed colonies without fluorescence and with the exception of two false negatives, these isolates were identified as non-C . albicans by other methods . CONCLUSIONS: Fluoroplate Candida allows a rapid and excellent identification of C . albicans showing a sensitivity and specificity of 99.3 and 99.7%, respectively.

J Dent Res, 1996 Dec, 75(12), 1979 - 85
Reactivity of Candida albicans germ tubes with salivary secretory IgA; Ponton J et al.; Salivary secretory IgA (sIgA) has been shown to react with a group of heat shock mannoproteins preferentially expressed on yeast cells grown at 37 degrees C . Since at this temperature C . albicans can induce germ tubes, we explored the role of germ tube induction on human salivary sIgA reactivity in both germinative and agerminative C . albicans strains, in an attempt to investigate whether the germ tube expressed the heat shock mannoproteins reactive with sIgA . The reactivity with sIgA of the agerminative strain, grown at 25 and 37 degrees C for different times, was measured spectrofluorometrically and was fairly constant with time . Yeast cells grown at 37 degrees C tended to be more reactive than those grown at 25 degrees C . In contrast, when compared with the yeast cells of the germinative strain grown at 25 degrees C, there was a statistically significant decrease in reactivity with sIgA during germ tube formation . Serum IgA and IgG did not show statistically significant changes in reactivity with C . albicans during germination, suggesting differences in reactivity with C . albicans cell wall antigens between mucosal and systemic humoral responses . Cell wall mannoproteins of molecular masses > 60 kDa were characterized by Western blotting as responsible for the decrease in sIgA reactivity observed in the germ tube, and the fall in sIgA reactivity was related to the release of cell wall mannoproteins into the culture medium . The release of these mannoproteins may be a mechanism whereby C . albicans avoids the action of sIgA, and it may play an important role in the post-parasite relationship in oral candidiasis.

Eur J Clin Microbiol Infect Dis, 1996 Dec, 15(12), 909 - 12
Occurrence of yeast bloodstream infections between 1987 and 1995 in five Dutch university hospitals; Voss A et al.; The aim of this study was to identify retrospectively trends in fungal bloodstream infections in The Netherlands in the period from 1987 to 1995 . Results of over 395,000 blood cultures from five Dutch university hospitals were evaluated . Overall, there were more than 12 million patient days of care during the nine-year study period . The rate of candidemia doubled in the study period, reaching an incidence of 0.71 episodes per 10,000 patient days in 1995 . The general increase in candidemia was paralleled by an increase in non-Candida albicans bloodstream infections, mainly due to Candida glabrata . However, more than 60% of the infections were caused by Candida albicans . Fluconazole-resistant species such as Candida krusei did not emerge during the study period . The increasing rate of candidemia found in Dutch university hospitals is similar to the trend observed in the USA, but the rate is lower and the increase is less pronounced.

J Antibiot (Tokyo), 1996 Dec, 49(12), 1221 - 5
Antifungal antibiotic benanomicin A increases susceptibility of Candida albicans to phagocytosis by murine macrophages; Watabe H et al.; Benanomicin A is an antifungal antibiotic produced by Actinomadura spadix . In the present study, we investigated the effect of benanomicin A on the phagocytosis of Candida albicans by murine peritoneal macrophages and on the cell-surface hydrophobicity (CSH) of C . albicans . Although pretreatment of macrophages with benanomicin A had no effect on the phagocytosis, addition of benanomicin A to the culture of macrophages and Candida cells increased the susceptibility of Candida cells to the phagocytosis by the macrophages . Pretreatment of Candida cells with benanomicin A also increased the susceptibility of Candida cells to the phagocytosis . When Candida cells were mixed with benanomicin A, the antibiotic bound irreversibly to Candida cells . These data suggest the possibility that the increased susceptibility of Candida cells to the phagocytosis is mediated by the binding of benanomicin A to Candida cells . Examination of physicochemical property of Candida cell surface showed that the CSH of Candida cells significantly decreased by the treatment with benanomicin A . Thus, binding of benanomicin A to Candida cells may induce biochemical/physicochemical alternation of the surfaces, so that they become more susceptible to phagocytosis by murine macrophages . These properties of benanomicin A, along with its antifungal activity, seem to be beneficial in the treatment of fungal infections.

J Antimicrob Chemother, 1996 Dec, 38(6), 953 - 61
Itraconazole for prophylaxis of systemic mycoses in neutropenic patients with haematological malignancies; Bohme A et al.; The efficacy of oral itraconazole 2 x 200 mg capsules daily for prevention of systemic mycoses was investigated in granulocytopenic patients with haematological malignancies . Of 241 patients, 197 were evaluable for prophylactic efficacy, and 214 for adverse events . Patients with similar characteristics receiving oral amphotericin B as antifungal prophylaxis, observed over 15 months before introduction of itraconazole, served as control group (n = 223) . With itraconazole prophylaxis, 13 cases of aspergillosis (9 proven, 1 probable, 3 possible; 7%) and no systemic yeast infection occurred, compared with 14 episodes of aspergillosis (9 proven, 2 probable, 3 possible; 6%) and 3 proven systemic yeast infections (Candida albicans, Candida norvegensis, Trichosporon beigelii) in the historical group . Adverse events were observed in 13% of evaluable patients receiving itraconazole . In four patients with acute lymphoblastic leukaemia receiving itraconazole and vincristine simultaneously, severe vinca alkaloid-induced neurotoxicity occurred . Plasma concentrations of itraconazole and hydroxyitraconazole were measured in 64 patients . After eight days of itraconazole the median drug concentration was adequate (700 ng/mL), but there was a marked individual variation (229-2861 ng/mL) . In comparison with a historical group, antifungal prophylaxis with itraconazole reduced the incidence of systemic yeast infections, but the frequency of aspergillosis was similar . However, a general increasing incidence of aspergillus infections at our hospital over the last four years should be considered in the assessment of study results.

Allergy, 1996 Dec, 51(12), 887 - 92
Allergenicity of acid protease secreted by Candida albicans; Akiyama K et al.; We have previously reported the cases of Candida albicans (C . alb) acid protease (CAAP)-induced atopic asthma . In this study, the allergenicity of the released enzyme CAAP was examined among asthmatic patients with positive immediate skin response to crude C . alb antigen . Among 49 patients with positive skin response to crude C . alb, anti-crude C . alb IgE antibodies were detected in 40 and anti-CAAP IgE antibodies were detected in 18 . Moreover, anticrude C . alb IgE antibodies were detected in all of the patients in whom anti-CAAP IgE antibodies were detected . No correlations between IgG antibodies to both antigens or between IgE and IgG antibodies to CAAP were observed . CAAP induced significant T-cell proliferation in 20/28 patients showing positive T-cell proliferation response to crude C . alb antigen . Most of the patients showing positive conjunctival response to crude C . alb antigen also showed positive response to CAAP . Most of the patients showing high levels of serum IgE antibody and positive histamine-release response of peripheral blood leukocytes to CAAP showed positive conjunctival response . The results indicate that CAAP is an important allergen in C . alb-related mucosal allergy.

Microbiologia, 1996 Dec, 12(4), 613 - 20
In vitro activity of fluconazole on Candida albicans; Abecia LC et al.; Fluconazole is a triazole antifungal compound suitable for the treatment of fungal infections, including those caused by Candida albicans . Fluconazole, as all azole antifungals, is a potent inhibitor of ergosterol biosynthesis . The aims of this study are to evaluate the susceptibility of C . albicans strains isolated from clinical specimens against fluconazole, and to assess the decrease in ergosterol produced on these strains when they are incubated in vitro with this antifungal compound . Sixty six yeast strains were isolated and identified from vaginal specimens of 710 women of a tocogynecology surgery (9.3%), C . albicans being the most frequent species (n = 52, ca . 79%) . An agar dilution technique was used to determine the minimal inhibitory concentration (MIC) of fluconazole for the C . albicans strains . The MICs rank was between 1 and 20 micrograms/ml (mean = 6.6 micrograms/ml) . Ergosterol content from ten C . albicans strains (MIC for fluconazole = 5 micrograms/ml) was assessed using the method proposed by Breivik and Owades, with three concentrations of fluconazole (2.5, 5 and 20 micrograms/ml) and four contact times (1, 6, 12 and 24 h), in comparison to no treated strains (control) . The mean content of ergosterol was lower in the treated strains than in the control ones, and became statistically significant after 12 h of incubation.

Drug Des Discov, 1996 Dec, 14(3), 171 - 8
A synthetic form of tracheal antimicrobial peptide has both bactericidal and antifungal activities; Lawyer C et al.; We have chemically synthesized tracheal antimicrobial peptide (TAP) with 38 amino acids and examined efficacy of the peptide on various organisms . The synthetic peptide showed potent bactericidal effect on both gram positive and negative bacteria . The action of bactericidal effect was relatively quick and 99.9% of E . coli cells were killed within 90 minutes at a concentration of 2.5 micrograms/ml of TAP . The peptide also showed antifungal activity against both mycelia (Aspergillus fumigatus) and yeast (Candida albicans) forms of fungi . Our domain analysis with a series of synthetic peptides of various lengths indicates that 17 amino acid residues of the C-terminal end is the minimum functional domain of the bactericidal activity.

Microbiology, 1996 Dec, 142 ( Pt 12), 3487 - 96
Neutrophil depletion increases susceptibility to systemic and vaginal candidiasis in mice, and reveals differences between brain and kidney in mechanisms of host resistance; Fulurija A et al.; Infections caused by the yeast Candida albicans represent an increasing threat to debilitated and immunosuppressed patients, and neutropenia is an important risk factor . Monoclonal antibody depletion of neutrophils in mice was used to study the role of these cells in host resistance . Ablation of neutrophils increased susceptibility to both systemic and vaginal challenge . The fungal burden in the kidney increased threefold on day 1, and 100-fold on day 4, and infection was associated with extensive tissue destruction . However, a striking feature of the disseminated disease in neutrophil-depleted animals was the altered pattern of organ involvement . The brain, which is one of the primary target organs in normal mice, was little affected . There was a threefold increase in the number of organisms recovered from the brains of neutrophil-depleted mice on day 4 after infection, but detectable abscesses were rare . In contrast, the heart, which in normal mice shows only minor lesions, developed severe tissue damage following neutrophil depletion . Mice deficient in C5 demonstrated both qualitative and quantitative increases in the severity of infection after neutrophil depletion when compared with C5-sufficient strains . The results are interpreted as reflecting organ-specific differences in the mechanisms of host resistance.

J Pharm Pharmacol, 1996 Dec, 48(12), 1315 - 9
Frequency distribution of Candida albicans blastospores adhered to mucosal epithelial cells in-vitro; Gorman SP et al.; Although several methods are available for examination of microbial adherence to epithelial cells, these do not distinguish between adherence of viable and non-viable micro-organisms . This study reports the use of acridine orange-stained blastospores of Candida albicans in conjunction with direct epifluorescence microscopy to determine viable (orange-fluorescing) and non-viable (green-fluorescing) blastospore adherence to buccal epithelial cells . The method was also employed to examine the effects of chlorhexidine treatment at subminimum inhibitory concentrations on the adherence of viable and non-viable blastospores . There was good correlation in the assessment of blastospore viability between the direct epifluorescence microscopy technique and the standard serial dilution and plating method for viable counting, confirming the reliability of direct epifluorescence microscopy . Chlorhexidine treatment before acridine orange staining did not alter this assessment of viability . Blastospore adherence to buccal epithelial cells resulted in a similarly skewed distribution whether examined using a crystal violet stain in conjunction with light microscopy or using direct epifluorescence microscopy, therefore validating the direct epifluorescence microscopy technique for the enumeration of blastospore adherence . Chlorhexidine treatment (0.0005% v/v, 30 min) of either blastospores or buccal epithelial cells altered the distribution of adherent blastospores per epithelial cell by increasing the number of epithelial cells having no adherent blastospores . No differences in adherence were, however, observed between blastospore or epithelial cells after treatment with this agent . Examination of the adherence of viable and non-viable blastospores to buccal epithelial cells using direct epifluorescence microscopy revealed a greater adherence capacity of non-viable than viable blastospores for buccal epithelial cells . Treatment of blastospores with chlorhexidine altered the frequency distributions of viable and non-viable blastospores with lower numbers of blastospores adherent per epithelial cell . The larger reduction in adherent viable blastospores in comparison with their non-viable counterparts is, however, an important observation which might have clinical relevance . Microbial cells adhere to epithelial cells resulting in a skewed distribution; study of this distribution gives useful information about the adherence process . Viable and non-viable components of a microbial population have different adherence capabilities and treatment of such populations with an antimicrobial agent exerting anti-adherent activity at sub-minimum inhibitory concentrations reduces the amount of adherence of these viable/non-viable components to different extents.

J Chemother, 1996 Dec, 8(6), 438 - 44
Interaction of Candida albicans, macrophages and fluconazole: in vitro and ex vivo observations; Tullio V et al.; In recent years, medical interest in evaluating the interaction among mycetes, phagocytes, and antimycotic drugs has increased notably due to higher incidences of fungal infections in immunocompromised subjects and to the long-term therapy they require . In this study the in vitro and ex vivo interaction of fluconazole, at plasma concentrations, with mouse macrophages was evaluated in the presence of different inocula of Candida albicans . The results showed that fluconazole did not interfere negatively with phagocyte functions; conversely, according to different experimental conditions, it was able to increase both phagocytosis and intracellular killing of candida, probably exerting its action on the yeast rather than on the phagocyte . A higher enhancement of macrophage functions was observed in vitro when the drug was present in the medium with macrophages and candida in a 1:1 ratio.

J Leukoc Biol, 1996 Dec, 60(6), 737 - 43
Infection by Candida albicans inhibits apoptosis of human monocytes and monocytic U937 cells; Heidenreich S et al.; Infectious microorganisms can differently induce or inhibit apoptosis of immunocompetent effector and host cells . In this study we examined the influence of an infection by Candida albicans (C . albicans) on programmed cell death of monocytic U937 cells and human monocytes . Basal and tumor necrosis factor alpha (TNF-alpha)-induced DNA fragmentation of U937 cells was significantly inhibited by an infection with C . albicans . Enhanced apoptosis of U937 cells, induced by TNF-alpha, caused a diminished candidacidal activity of the effector cells, whereas inhibition of apoptosis by granulocyte-macrophage colony-stimulating factor (GM-CSF) was paralleled by an intensified host defense . Pretreatment of U937 cells or monocytes with the cyclooxygenase blocker indomethacin completely abolished the reduction of DNA fragmentation induced by the yeast . Studying the underlying mechanisms we found that C . albicans induced formation of prostaglandin E2 (PGE2) by U937 . Exogenous administration of PGE2 down-regulated apoptosis of U937 or human monocytes to a similar extent as did fungal infection . Activation of protein kinase A by the cAMP analogue 8-bromo-cAMP inhibited U937 apoptosis, as did PGE2 . On the other hand, rp-cAMP, a blocker of the cAMP-dependent signal transduction, restored and elevated DNA fragmentation levels down-regulated by C . albicans . U937 cells expressed the bcl-2 protein but the infection with fungi or PGE2 treatment did not increase proto-oncogene expression . Monocytic effector cells may therefore strengthen the defense against C . albicans by an autocrine feedback regulation via a PGE2-dependent, cAMP-transduced inhibition of apoptosis.

FEMS Microbiol Lett, 1996 Dec 1, 145(2), 167 - 72
Ultrastructural and biochemical studies on Candida albicans after prolonged incubation in sea water; Gorga F et al.; Candida albicans yeast cells suspended in sterilized sea water and cultivated in Brain Heart Infusion broth were compared . Viability, chemical composition, surface hydrophobicity and ultrastructural characteristics showed variations after incubation in sea water . The yeast cells developed some ultrastructural changes after about a month in sea water . The surface hydrophobicity of the yeast cells was gradually reduced, starting from day 16, and continued to decline throughout the 32 days in sea water . A decrease in total carbohydrate, lipid and protein contents was also observed and corresponded with ultrastructural modifications.

FEMS Microbiol Lett, 1996 Dec 1, 145(2), 157 - 62
Molecular cloning and characterization of a Candida albicans gene (EFB1) coding for the elongation factor EF-1 beta; Maneu V et al.; A Candida albicans gene homologous to Saccharomyces cerevisiae elongation factor 1 beta was isolated by screening a genomic DNA library using a C . albicans cDNA as a probe . This cDNA was previously obtained by immunoscreening of an expression library with polyclonal antibodies raised against candidal cell wall components . Sequence analysis of the cDNA and the whole C . albicans gene (EMBL accession number X96517) revealed an intron-interrupted open reading frame of 639 base pairs that encodes a 213 amino acid protein . Exon sequences are highly homologous (74%) to S . cerevisiae EFB1, whereas intron sequence is less conserved (34% identity), and the predicted amino acid sequence shares about 73% identity.

Hum Gene Ther, 1996 Dec 1, 7(18), 2255 - 61
Transfer of a gene encoding the anticandidal protein histatin 3 to salivary glands; O'Connell BC et al.; Mucosal candidiasis, the most common opportunistic fungal infection in human immunodeficiency virus (HIV)-infected patients, is an early sign of clinically overt acquired immunodeficiency syndrome (AIDS) and an important cause of morbidity, particularly in HIV-infected children . The appearance of azole-resistant strains of Candida albicans had made clinical management of candidiasis increasingly difficult . We propose a novel approach to the management of candidal infections that involves the use of naturally occurring antifungal proteins, such as the histatins . Histatins are a family of small proteins that are secreted in human saliva . We have constructed recombinant adenovirus vectors that contain the histatin 3 cDNA . These vectors are capable of directing the expression of histatin 3 in the saliva of rats at up to 1,045 micrograms/ml, well above the levels found in normal human saliva . The adenovirus-directed histatin demonstrated a 90% candidacidal effect in the timed-kill assay against both fluconazole-susceptible and fluconazole-resistant strains of C . albicans and inhibited germination by 45% in the same strains . These studies suggest that a gene transfer approach to overexpress naturally occurring antifungal proteins may be useful in the management of mucosal candidiasis.

Infect Immun, 1996 Dec, 64(12), 5092 - 7
Importance of beta2-microglobulin in murine resistance to mucosal and systemic candidiasis; Balish E et al.; beta2-Microglobulin knockout (beta2m-/-) mice, which lack major histocompatibility complex class I expression and are deficient in CD8alpha/beta T-cell receptor alpha/beta (TcRalpha/beta) T cells, were as resistant to systemic (intravenous) challenge with Candida albicans as immunocompetent controls . Conversely, the beta2m-/- mutant mice were susceptible to systemic candidiasis of endogenous origin despite the induction of C . albicans-specific antibody and cell-mediated immune responses after colonization with a pure culture of C . albicans . Despite some superficial and transient infections of tongues and esophagi (detected by histology) at 1 to 2 weeks after oral colonization and gastric infections (cardia-antrum section) which were observed at 10 to 12 weeks after oral challenge, C . albicans-colonized beta2m-/- mice showed an overall resistance to candidiasis in other mucosal and cutaneous tissues . These data suggest that immune defects that accompany the loss of beta2-microglobulin play an important role in murine resistance to gastric and disseminated candidiasis of endogenous (intestinal tract) origin and that innate immunity and CD4 TcRalpha/beta as well as CD8alpha/alpha TcRalpha/beta (or -gamma/delta) T cells play an important role in resistance to systemic, cutaneous, and nongastric mucosal tissues.

Infect Immun, 1996 Dec, 64(12), 5085 - 91
Migration of the fungal pathogen Candida albicans across endothelial monolayers; Zink S et al.; Migration of the fungal pathogen Candida albicans across the endothelial cell layer is considered a prerequisite for the invasion of multiple organs occurring in systemic candidiasis . We developed an experimental system in which C . albicans migrates from a luminal compartment across a monolayer of bovine aortic endothelial cells on a porous filter support to an abluminal compartment . In this system, a C . albicans wild-type strain (ATCC 10261) traverses the endothelial monolayer in a time-, glucose-, and cell concentration-dependent manner . A mutant derivative unable to grow and form hyphae (SGY-243) migrates at a reduced rate . Concomitant to transendothelial migration, the permeability of the endothelial monolayer for dextran diffusion markers is significantly increased . This increase in transendothelial exchange occurs before fungal cells are detectable in the abluminal compartment and is time, glucose, and cell concentration dependent . A mutant strain (hOG301) unable to interact with endothelial cells does not alter endothelial permeability . Thus, transendothelial migration of C . albicans is able to damage the barrier function of an endothelial monolayer . Our experimental system, which reflects key stages of transendothelial migration of C . albicans including adherence and passage across endothelial cells and the extracellular matrix, may be a useful model for comparisons of transendothelial migration characteristics of Candida strains.

Infect Immun, 1996 Dec, 64(12), 5000 - 7
Candidacidal activity of recombinant human salivary histatin-5 and variants; Tsai H et al.; Human salivary histatins possess fungicidal and bactericidal activities . The current investigation evaluates the structure-function relationship of histatins with regard to their candidacidal activity by using recombinant histatin-5 and its variants produced in Escherichia coli . The purified recombinant histatins were examined for their candidacidal activity and secondary structure . The m21 (with Lys-13 replaced by Thr {Lys-13-->Thr}) and m71 (Lys-13-->Glu) variants are significantly less effective than recombinant histatin-5 in killing Candida albicans, suggesting that Lys-13 is critical for candidacidal activity . The m68 (Lys-13-->Glu and Arg-22-->Gly) variant is significantly less potent than the recombinant histatin-5 as well as m71, indicating that Arg-22 is crucial for the cidal activity . The candidacidal activities of m1 (Arg-12-->Ile), m2 (Arg-12-->Ile and Lys-17-->Asp), m12 (Arg-12-->Lys and His-21-->Leu), and m70 (His-19-->Pro and His-21-->Arg) variants, however, are comparable to that of recombinant histatin-5, indicating that Arg-12, Lys-17, His-19, and His-21 are not functionally important . The conformational preferences of histatin-5 and variants were determined by circular dichroism . The results indicate that all proteins have a strong tendency to adopt alpha-helical conformation in trifluoroethanol . Previously, we have shown that the alpha-helical conformation is one of the important structural requirements for eliciting appreciable candidacidal activity . Collectively, the data suggest that in addition to the helical conformation, specific residues such as Lys-13 and Arg-22 in the sequence of histatin-5 are, indeed, important for candidacidal activity.

Infect Immun, 1996 Dec, 64(12), 4907 - 14
CD4+ T-helper-cell responses in mice with low-level Candida albicans infection; Mencacci A et al.; Resistance and susceptibility to Candida albicans infection have been shown to be dependent upon the activation of CD4+ T helper (Th) type 1 or Th2 cells, respectively . To study the type, kinetics, and cytokine dependency of CD4+ Th-cell responses in low-level C . albicans infection, susceptible mice were infected with sublethal doses of C . albicans and assessed for parameters of CD4+ Th-dependent immunity . Interleukin (IL)-12 and gamma interferon were always produced early in infection regardless of the pathogen load . In contrast, production of IL-4, and hence Th2-cell reactivity, was strictly dose dependent, being induced at the higher dose of the fungus . Production of IL-12 correlated with a successful control of infection in mice exposed to the lower doses of C . albicans but not with the development of acquired immunity . An antigenic stimulus appeared to be required for IL-12 to induce a protective anticandidal response . Cytokine depletion in vivo revealed that neutralization of IL-4 was protective early but not late in infection, suggesting a different role for IL-4 in the induction versus maintenance of an ongoing anticandidal Th response . Late in infection, an exacerbative effect was also observed upon IL-12 neutralization . These results indicate that the fungal burden and timing of cytokine appearance greatly influence CD4+ Th induction and effector functions in mice with candidiasis.

J Clin Microbiol, 1996 Dec, 34(12), 3237 - 9
Comparative evaluation of macrodilution and chromogenic agar screening for determining fluconazole susceptibility of Candida albicans; Patterson TF et al.; A simple screening method for fluconazole susceptibility using CHROMagar Candida with fluconazole was compared with the National Committee for Clinical Laboratory Standards (NCCLS) macrobroth method . In this agar dilution method, susceptible Candida albicans colonies are smaller on medium with fluconazole than on fluconazole-free medium . Yeasts with decreased susceptibility have normal-sized colonies on medium containing fluconazole . On agar with 16 micrograms of fluconazole per ml, 32 of 34 strains with NCCLS MICs of > or = 16 micrograms/ml were correctly predicted, as were 66 of 68 with MICs of < 16, an agreement of 96% . On agar with 8 micrograms of fluconazole per ml, 38 of 41 isolates with MICs of > or = 8 were correctly predicted, as were 59 of 61 isolates with MICs of < 8, an agreement of 95% . This agar dilution methods appears to highly correlate with NCCLS macrobroth methods for detection of C . albicans and may be an effective screen for fluconazole susceptibility.

J Clin Microbiol, 1996 Dec, 34(12), 3208 - 11
Comparative evaluation of three antifungal susceptibility test methods for Candida albicans isolates and correlation with response to fluconazole therapy; Ruhnke M et al.; In vitro susceptibilities were determined for 56 Candida albicans isolates obtained from the oral cavities of 41 patients with human immunodeficiency virus infection . The agents tested included fluconazole, itraconazole, ketoconazole, flucytosine, and amphotericin B . MICs were determined by the broth microdilution technique following National Committee for Clinical Laboratory Standards document M27-P (M27-P micro), a broth microdilution technique using high-resolution medium (HR micro), and the Etest with solidified yeast-nitrogen base agar . The in vitro findings were correlated with in vivo response to fluconazole therapy for oropharyngeal candidiasis . For all C . albicans isolates from patients with oropharyngeal candidiasis not responding to fluconazole MICs were found to be > or = 6.25 micrograms/ml by the M27-P micro method and > or = 25 micrograms/ml by the HR micro method as well as the Etest . However, for several C . albicans isolates from patients who responded to fluconazole therapy MICs found to be above the suggested breakpoints of resistance . The appropriate rank order of best agreement between the M27-P micro method and HR micro method was amphotericin B > fluconazole > flucytosine > ketoconazole > itraconazole . The appropriate rank order with best agreement between the M27-P micro method and the Etest was flucytosine > amphotericin B > fluconazole > ketoconazole > or = itraconazole . It could be concluded that a good correlation between in vitro resistance and clinical failure was found with all methods . However, the test methods used in this study did not necessarily predict clinical response to therapy with fluconazole.

J Clin Microbiol, 1996 Dec, 34(12), 3040 - 3
Performance of fungal blood cultures by using the Isolator collection system: is it cost-effective?
Morrell RM Jr, Wasilauskas BL, Steffee CH.
Enhanced recovery of fungal isolates from blood by using the Isolator system has been reported previously . We examined bacterial and fungal blood cultures during a 14-month period to determine if this enhanced recovery required a separate fungal culture and to determine the differential utility between a fungal blood culture and a routine bacterial culture . During this period, 84 of 5,196 (1.6%) fungal blood cultures and 170 of 25,702 (0.6%) bacterial blood cultures were positive for yeast or filamentous fungi . Thirty-seven positive fungal cultures, simultaneously collected, had correspondingly positive bacterial cultures . An additional 15 positive fungal cultures yielded isolates that had either been previously recovered from a bacterial culture or were recovered from a bacterial culture collected within 48 h . Of the 32 unpaired fungal cultures remaining, 5 were Candida albicans whose unique isolation was believed to be the result of specimen sampling variance rather than any enhanced recovery characteristics of fungal culture methods . Examination of patient data relating to the 27 remaining isolates (24 patients episodes) showed that only five fungal blood cultures (0.096% of all collected) had any impact on patient therapy decisions, and one of these was judged to be the cause of unnecessary therapy . Our data suggest that separate fungal cultures of blood are not cost-effective for those laboratories using the Isolator for routine blood cultures and furthermore may not be cost-effective for laboratories using automated broth systems that are comparable to the Isolator in recovery of fungiPublication Types:

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