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 subse