Gene, 1998 Oct 5, 220(1-2), 91 - 8 A fourth gene from the Candida albicans CDR family of ABC transporters; Franz R et al.; Using primers derived from a region of the Candida albicans CDR1 (Candida drug resistance) gene that is conserved in other ABC (ATP-binding cassette) transporters, a DNA fragment from a previously unknown CDR gene was obtained by polymerase chain reaction (PCR) . After screening a C . albicans genomic library with this fragment as a probe, the complete CDR4 gene was isolated and sequenced . CDR4 codes for a putative ABC transporter of 1490 amino acids with a high degree of homology to Cdr1p, Cdr2p and Cdr3p from C . albicans (62, 59 and 57% amino acid sequence identity, respectively) . Cdr4p has a predicted structure typical for cluster I . 1 of yeast ABC transporters, characterized by two homologous halves, each comprising an N-terminal hydrophilic domain with consensus sequences for ATP binding and a C-terminal hydrophobic domain with six transmembrane helices . In contrast to the CDR1/CDR2 genes, the genetic structure of the CDR4 gene was conserved in 59 C . albicans isolates from six different patients . Northern hybridization analysis showed that the CDR4 gene was expressed in most isolates, but no correlation between CDR4 mRNA levels and the degree of fluconazole resistance of the isolates was found . In addition, a C . albicans mutant in which both copies of the CDR4 gene were disrupted by insertional mutagenesis was not hypersusceptible to fluconazole as compared to the parent strain . Unlike CDR1 and CDR2, CDR4 does not, therefore, seem to be involved in fluconazole resistance in C . albicans. Res Microbiol, 1998 May, 149(5), 327 - 38 Reaggregation and binding of cell wall proteins from Candida albicans to structural polysaccharides; Aguado C et al.; Urea or hot sodium dodecyl sulphate extracted a significant amount of the same proteins from the matrix of the cell wall of the yeast form and mycelial cells of Candida albicans . Gel filtration analysis of the urea-extracted proteins revealed that they occurred in the form of large complexes which were unaffected by up to 8 M urea . Among them, proteins en route to becoming covalently associated within the wall scaffold were identified by their reaction with specific antibodies . When urea was removed by dialysis, some of these proteins specifically reassociated into large aggregates which bound strongly with ConA, whereas others remained soluble in smaller associated products . The ability of some of these proteins to bind to the insoluble wall polysaccharides was also assessed . No self-assembling proteins were able to bind to glucans and/or chitin . Specificity of the binding to polysaccharides made of beta-bound glucosyl or N-acetylglucosaminyl residues was determined by the competitive effect of several disaccharides . Whereas laminaribiose and diacetylchitobiose were strong inhibitors of protein binding to both glucan and chitin, lactose, maltose and sucrose were ineffective. Res Microbiol, 1997 Sep-Oct, 148(7), 593 - 603 Initial steps of wall protoplast regeneration in Candida albicans; Rico H et al.; Cell wall regeneration of individual Candida albicans yeast and mycelial protoplasts was studied with confocal and electron microscopy using polyclonal antibodies and lectins . Quantitative measurements of the fluorescence emitted by individual protoplasts during the process of regeneration indicate that chitin is the first polymer to be laid down, whereas beta (1,3)- and beta (1,6)glucan are incorporated at a later stage . Mannoproteins were found on the surface of fresh protoplasts and those newly synthesized were then deposited with time . During the first steps of wall regeneration, the proteins that interacted covalently with chitin or glucan were different, but the same species were found linked to each polymer in yeast and mycelial regenerating forms . The aggregates formed by regenerating protoplasts were shown to be due to the chitin and mannoprotein network initially laid. J Bacteriol, 1998 Oct, 180(20), 5334 - 43 Identification of Candida albicans ALS2 and ALS4 and localization of als proteins to the fungal cell surface; Hoyer LL et al.; Additional genes in the growing ALS family of Candida albicans were isolated by PCR screening of a genomic fosmid library with primers designed from the consensus tandem-repeat sequence of ALS1 . This procedure yielded fosmids encoding ALS2 and ALS4 . ALS2 and ALS4 conformed to the three-domain structure of ALS genes, which consists of a central domain of tandemly repeated copies of a 108-bp motif, an upstream domain of highly conserved sequences, and a domain of divergent sequences 3' of the tandem repeats . Alignment of five predicted Als protein sequences indicated conservation of N- and C-terminal hydrophobic regions which have the hallmarks of secretory signal sequences and glycosylphosphatidylinositol addition sites, respectively . Heterologous expression of an N-terminal fragment of Als1p in Saccharomyces cerevisiae demonstrated function of the putative signal sequence with cleavage following Ala17 . This signal sequence cleavage site was conserved in the four other Als proteins analyzed, suggesting identical processing of each protein . Primary-structure features of the five Als proteins suggested a cell-surface localization, which was confirmed by indirect immunofluorescence with an anti-Als antiserum . Staining was observed on mother yeasts and germ tubes, although the intensity of staining on the mother yeast decreased with elongation of the germ tube . Similar to other ALS genes, ALS2 and ALS4 were differentially regulated . ALS4 expression was correlated with the growth phase of the culture; ALS2 expression was not observed under many different in vitro growth conditions . The data presented here demonstrate that ALS genes encode cell-surface proteins and support the conclusion that the size and number of Als proteins on the C . albicans cell surface vary with strain and growth conditions. Int J STD AIDS, 1998 Sep, 9(9), 526 - 30 Vulvovaginal candidiasis in female sex workers; Otero L et al.; Vulvovaginal candidiasis is a frequent inflammatory process in women but it has not been widely studied in female sex workers (FSWs) . To estimate the frequency of Candida species infection in FSWs and to identify related risk factors and clinical findings, we carried out a retrospective study of 1923 FSWs over 11 years . We also performed a prospective study of 163 consecutive FSWs with a history of candidiasis during a 4-year period . Candida species were isolated in 1967 samples (18.5% of the total) . Candida albicans (89.3%) was the most frequent species, followed by Candida glabrata (2.7%), Candida parapsilosis (1.2%) and Saccharomyces cerevisiae (0.4%) . In the prospective study of 163 patients, we found vaginal discharge in 76.1% of cases, soreness in 52.1% and vulval pruritus in 32.5% . We identified 12 patients (7.4%) with recurrent vulvovaginal candidiasis . No statistical difference was found between recurrent vulvovaginitis and the use of oral contraceptives, oral sex, tight-fitting clothing and synthetic underwear . FSWs have the same prevalence of candidiasis as other groups of women described in published literature . The proportion of albicans and non-albicans species does not differ between women with recurrent and non-recurrent vulvovaginal candidiasis (VVC). AIDS, 1998 Sep 10, 12(13), 1601 - 10 Change in fluconazole susceptibility patterns and genetic relationship among oral Candida albicans isolates; Diaz-Guerra TM et al.; OBJECTIVE: To assess the genetic homogeneity or heterogeneity within each set of Candida albicans isolates colonizing/infecting the oral cavities of HIV-infected patients undergoing azole therapy when changes in susceptibility to fluconazole were detected . DESIGN: Fourteen HIV-positive patients suffering recurrent episodes of oral candidosis were prospectively followed from the first episode to the isolation of strains with decreased susceptibility to fluconazole . The strains of C . albicans isolated either from episodes or controls throughout the prospective study were analysed . METHODS: Electrophoretic karyotyping and hybridization with the repeated sequence probe 27A were used to delineate sequential isolates . In vitro susceptibility tests to fluconazole and ketoconazole were also performed . The results obtained by DNA fingerprinting with the probe combined with computer-assisted analysis were used to assess the genetic relationships amongst the strains . In addition, comparison with the genetic relatedness of a group of geographically unrelated strains was made . RESULTS: Isogenic populations of sequential isolates were observed only in two patients; 12 patients harboured heterogenic populations over time, although in 11 patients there was a predominant strain that was isolated more than once, and only one of these patients carried strains with a similarity index less than 80% . With the exception of two patients, each patient carried a major strain that became less susceptible to fluconazole . The similarity index for the unrelated strains was 59% . CONCLUSIONS: HIV-infected patients may carry a mixed population of strains, but the strains tend to be related to each other . The strains were maintained throughout the course of infection and at least one developed secondary resistance to fluconazole. Farmaco, 1998 Jun 30, 53(6), 415 - 20 Synthesis of some new benzimidazolecarboxamides and evaluation of their antimicrobial activity; Goker H et al.; A series of 1,2-disubstituted benzimidazole-5(6)-carboxamides was prepared and evaluated in vitro for antimicrobial activity against Staphyloccus aureus, Escherichia coli and Candida albicans . The precursor benzimidazolecarboxylic acids 4a-c and 9a-c were prepared via oxidative condensation of diaminobenzoic acids with aldehydes and via several steps over the 2(1H)-benzimidazolones, respectively . All acids were converted to their acyl chlorides with SOCl2, then amidified with several N,N'-dialkylaminoethyl derivatives . Compounds 8a-c, 20 and 22 exhibited the best activity. Exp Gerontol, 1998 Aug, 33(5), 477 - 84 Age-associated differences in neutrophil oxidative burst (chemiluminescence); Braga PC et al.; Phagocytic defensive functions consist of a sequence of events, including migration, phagocytosis, secretion, and the release of reactive oxygen species (ROS) . The last of these (also called "oxidative burst") has not received due attention in the elderly, even though it can be considered the most important event in the process of killing an invading microorganism . The aim of the present study was to investigate the oxidative burst activity of polymorphonuclear neutrophil leukocytes (PMNs) in relation to age, using a technique that specifically identifies ROS production: luminol-amplified chemiluminescence (LACL) . Besides the use of LACL, a particular feature of the study was the use of five rather than just one or two different stimulants: two particulate (Candida albicans and zymosan) and three soluble ones {N-formyl-methionyl-leucyl-phenylalanine (fMLP), phorbol 12 myristate 13 acetate (PMA), and polyanetholesulfonate (liquoid)} . This approach allowed us to observe a dichotomy between the effects of Candida and zymosan (particulates), which were not significantly different in the elderly subjects compared to the young controls, and those of fMLP, PMA, and liquoid (solubles), which showed a significant reduction in LACL in the elderly group . Considering the different results obtained with the various stimulants adopted that are all believed to have NADPH oxidase as a common final target of oxidative burst, it may be postulated that aging can influence the different transductional pathways in different ways. Otolaryngol Pol, 1998, 52(3), 277 - 80 {The role of imperfect fungi in etiopathogenesis of allergic rhinitis}; Namyslowski G et al.; In this paper the role of imperfect fungi in etiopathogenesis of perennial rhinitis was examined . In a group of 26 patients the concentration of total IgE and IgE specific of Candida albicans, Aspergillus fumigatus, Alternaria alternata, Mucor racemosus and Cladosporum herbarum was signified . Oversensibility to imperfect fungi was confirmed in 30.8% of patients: to Candida albicans in 3.8%, to Aspergillus fumigatus in 11.5%, to Alternaria alternata in 3.8%, to Mucor racemosus in 7.6% and to Cladosporum herbarum in 3.8%. Biochemistry, 1998 Oct 6, 37(40), 14326 - 36 Binding and reactivity of Candida albicans estrogen binding protein with steroid and other substrates; Buckman J et al.; In this report recombinant estrogen binding protein (EBP1), isolated originally from Candida albicans as a result of its high affinity for 17beta-estradiol, has been purified extensively using a modified affinity purification scheme originally developed for a homolog of EBP1, old yellow enzyme (OYE) . It is shown that like OYE, the protein binds a variety of compounds with a phenolic structure, including 17beta-estradiol, and compounds with an alpha, beta-unsaturated keto or aldehyde structure . In addition, EBP1 exhibits an NADPH oxidoreductase activity, transferring electrons from NADPH to all alpha,beta-unsaturated ketones and aldehydes tested via the tightly bound FMN cofactor . Analysis of the steady-state kinetics of these reactions indicate a tetra uni ping-pong mechanism . Inhibition of the steady-state reaction by 17beta-estradiol gives a Ki = 10 +/- 2 nM, and indicates exclusive binding of this steroid to the enzyme in its oxidized state . In contrast, 19-nortestosterone binds to both oxidized and reduced forms of the enzyme with dissociation constants of 600 +/- 100 and 650 +/- 90 nM, respectively . EBP1 also catalyzes a disproportionation reaction with certain compounds, in which two molecules of a cylic alpha,beta-unsaturated ketone, including the steroid 19-nortestosterone, are individually aromatized and reduced to the corresponding saturated ketone . Despite the extensive similarity in sequence and enzymic activity, notable differences between EBP1 and the OYE family of proteins exist with regard to the binding behavior and reactivity with the two steroids tested here, estradiol and 19-nortestosterone. J Immunol, 1998 Oct 1, 161(7), 3543 - 50 IFN-gamma is required for IL-12 responsiveness in mice with Candida albicans infection; Cenci E et al.; To elucidate the role of IFN-gamma in antifungal CD4+ Th-dependent immunity, 129/Sv/Ev mice deficient for IFN-gamma receptor (IFN-gammaR(-/-)) were assessed for susceptibility to gastrointestinal or systemic Candida albicans infection and for parameters of innate and adaptive T helper immunity . IFN-gammaR(-/-) mice failed to mount protective Th1-mediated acquired immunity upon mucosal immunization or in response to a live vaccine strain of the yeast . The impaired Th1-mediated resistance correlated with defective IL-12 responsiveness, but not IL-12 production, and occurred in the presence of an increased innate antifungal resistance . The development of nonprotective Th2 responses was observed in IFN-gammaR(-/-) mice upon mucosal infection and subsequent reinfection . However, under experimental conditions of Th2 cell activation, the occurrence of Th2 cell responses was similar in IFN-gammaR(-/-) and in IFN-gammaR(+/+) mice . These results indicate the complex immunoregulatory role of IFN-gamma in the induction of mucosal and nonmucosal anticandidal Th cell responses; IFN-gamma is not essential for the occurrence of Th2 responses but is required for development of IL-12-dependent protective Th1-dependent immunity. Ann Allergy Asthma Immunol, 1998 Sep, 81(3), 247 - 55 IgE-sensitization to cellular and culture filtrates of fungal extracts in patients with atopic dermatitis; Nissen D et al.; BACKGROUND: Patients with atopic dermatitis may experience exacerbations of eczema triggered by various inflammatory stimuli . One mechanism may be IgE-mediated reactions to dermatophytes since these patients are more likely to acquire skin infections with dermatophytes and may become sensitized . OBJECTIVE: This study investigates IgE-sensitization to fungi in patients with atopic dermatitis and compares the biologic activity of culture filtrates and cellular fungal extracts . The following allergen extracts were provided as culture filtrates and cellular extracts: Candida albicans, Fusarium moniliforme, and Penicillium notatum . In addition, Pityrosporum ovale and Trichophyton rubrum cultures were included in the test panel . METHODS: Fifteen patients with clinical findings suggesting dermatophytosis and 11 controls were selected . Each subject was tested by leukocyte histamine release and skin prick test to each fungal extract . The extracts were separated and reduced by sodium dodecylsulfate polyacrylamide gel electrophoresis and analyzed by IgE-immunoblotting with sera from all study subjects . RESULTS: Fourteen patients (93%) reacted to one or several fungal extracts by releasing histamine when challenged in vitro . By immunoblotting experiments, patient sera showed binding to a wide range of components in all extracts . Patient sera recognized allergenic components shared by culture filtrates and cellular extracts but with higher frequent and greater intensity in culture filtrates . Although culture filtrates generated more frequent and potent IgE-reactions than the cellular extracts, the difference was not statistically significant . Biologic potency was similar when evaluated by skin prick tests and leukocyte histamine release . CONCLUSION: Patients with atopic dermatitis may develop specific IgE-antibodies to a number of fungi as demonstrated by IgE-immunoblotting . In selected patients, fungi may trigger an IgE-mediated reaction that may contribute to the exacerbation of eczema . Approximately, one-half of the patients, however, produced IgE-antibodies to fungal (glyco)proteins without a significant histamine release or skin test response possibly because of nonspecific interaction with carbohydrate moieties on IgE and poor biologic activity of IgE antibodies directed to cross-reactive carbohydrate determinants of fungal glycoproteins . This warrants caution when interpreting clinical relevance of serologic measurements of fungal IgE-antibodies. Antimicrob Agents Chemother, 1998 Oct, 42(10), 2745 - 6 Enhancement of antimicrobial activity of neuropeptide Y by N-terminal truncation; Shimizu M et al.; The activity of neuropeptide Y (NPY) against Candida albicans, which was revealed to be fungicidal, was enhanced significantly by the truncation of amino acid residues at the N terminus . The most active peptides (MICs, approximately 1 microM) were about 10-fold more potent than the intact NPY (MIC, approximately 10 microM) . The enhancement was weakened by the replacement of the N terminus by negatively charged residues and/or acylation of the alpha-amino group . These results suggest that only the alpha-helical region of NPY is necessary for the antimicrobial activity and that the net charge of the peptide is important for the activity. Antimicrob Agents Chemother, 1998 Oct, 42(10), 2645 - 9 Mechanism of fluconazole resistance in Candida krusei; Orozco AS et al.; The mechanisms of fluconazole resistance in three clinical isolates of Candida krusei were investigated . Analysis of sterols of organisms grown in the absence and presence of fluconazole demonstrated that the predominant sterol of C . krusei is ergosterol and that fluconazole inhibits 14alpha-demethylase in this organism . The 14alpha-demethylase activity in cell extracts of C . krusei was 16- to 46-fold more resistant to inhibition by fluconazole than was 14alpha-demethylase activity in cell extracts of two fluconazole-susceptible strains of Candida albicans . Comparing the carbon monoxide difference spectra of microsomes from C . krusei with those of microsomes from C . albicans indicated that the total cytochrome P-450 content of C . krusei is similar to that of C . albicans . The Soret absorption maximum in these spectra was located at 448 nm for C . krusei and at 450 nm for C . albicans . Finally, the fluconazole accumulation of two of the C . krusei isolates was similar to if not greater than that of C . albicans . Thus, there are significant qualitative differences between the 14alpha-demethylase of C . albicans and C . krusei . In addition, fluconazole resistance in these strains of C . krusei appears to be mediated predominantly by a reduced susceptibility of 14alpha-demethylase to inhibition by this drug. Antimicrob Agents Chemother, 1998 Oct, 42(10), 2584 - 9 Rapid, transient fluconazole resistance in Candida albicans is associated with increased mRNA levels of CDR; Marr KA et al.; Fluconazole-resistant Candida albicans, a cause of recurrent oropharyngeal candidiasis in patients with human immunodeficiency virus infection, has recently emerged as a cause of candidiasis in patients receiving cancer chemotherapy and marrow transplantation (MT) . In this study, we performed detailed molecular analyses of a series of C . albicans isolates from an MT patient who developed disseminated candidiasis caused by an azole-resistant strain 2 weeks after initiation of fluconazole prophylaxis (K . A . Marr, T . C . White, J . A . H . vanBurik, and R . A . Bowden, Clin . Infect . Dis . 25:908-910, 1997) . DNA sequence analysis of the gene (ERG11) for the azole target enzyme, lanosterol demethylase, revealed no difference between sensitive and resistant isolates . A sterol biosynthesis assay revealed no difference in sterol intermediates between the sensitive and resistant isolates . Northern blotting, performed to quantify mRNA levels of genes encoding enzymes in the ergosterol biosynthesis pathway (ERG7, ERG9, and ERG11) and genes encoding efflux pumps (MDR1, ABC1, YCF, and CDR), revealed that azole resistance in this series is associated with increased mRNA levels for members of the ATP binding cassette (ABC) transporter superfamily, CDR genes . Serial growth of resistant isolates in azole-free media resulted in an increased susceptibility to azole drugs and corresponding decreased mRNA levels for the CDR genes . These results suggest that C . albicans can become transiently resistant to azole drugs rapidly after exposure to fluconazole, in association with increased expression of ABC transporter efflux pumps. Antimicrob Agents Chemother, 1998 Oct, 42(10), 2534 - 41 Identification and characterization of novel antimicrobial decapeptides generated by combinatorial chemistry; Hong SY et al.; Novel combinatorial libraries consisting of simplified amino acid sequences were designed to screen for peptides active against the Candida albicans membrane . A novel decapeptide, KKVVFKVKFK, that had a unique primary amino acid sequence was identified in this work . This peptide irreversibly inhibited the growth of C . albicans and showed a broad range of antibacterial activity but no hemolytic activity . Circular dichroism spectra revealed that the predominant secondary structure of this peptide strongly depended on the membrane-mimetic environments; the peptide preferred to form an amphipathic alpha-helical structure in the presence of 50% trifluoroethanol, while it preferred to adopt a distorted alpha-helical structure in the presence of sodium dodecyl sulfate micelles . Experiments in which dye was released from vesicles indicated that this novel antimicrobial peptide killed microorganisms through the action on the membrane as its primary target . Replacement of amino acids in this active decapeptide on the basis of information from the libraries could provide unique information about factors affecting its antimicrobial activity such as its secondary structure, net positive charge, and hydrophobicity. Chemotherapy, 1998 Nov-Dec, 44(6), 405 - 8 Effects of cefepime, cefixime and ceftibuten on murine gut colonization by Candida albicans; Maraki S et al.; Crl:CD1(ICR) BR mice were fed chow containing Candida albicans or regular chow . Both groups were subsequently given either antibiotics or normal saline . Stool cultures were performed before, at the end of treatment and 1 week after treatment, to determine the effect on the stool yeast concentration . Candida-colonized mice treated with cefepime, cefixime or ceftibuten had higher (however not significantly) counts of the yeast in their stools than control Candida-fed mice treated with saline . A group of Candida-fed mice were treated with ceftriaxone, which is known to increase the yeast stool concentration significantly and served as positive control . Mice fed regular chow and treated with the study drugs or saline did not have any yeasts in their stools . Dissemination of Candida did not occur. FEBS Lett, 1998 Sep 11, 435(1), 49 - 54 Isolation and characterization of the Candida albicans gene for mRNA 5'-triphosphatase: association of mRNA 5'-triphosphatase and mRNA 5'-guanylyltransferase activities is essential for the function of mRNA 5'-capping enzyme in vivo; Yamada-Okabe T et al.; The amino acid sequence of the Saccharomyces cerevisiae mRNA 5'-triphosphatase (TPase) diverges from those of higher eukaryotes . In order to confirm the sequence divergence of TPases in lower and higher eukaryotes, the Candida albicans gene for TPase was identified and characterized . This gene designated CaCET1 (C . albicans mRNA 5'-capping enzyme triphosphatase 1) has an open reading frame of 1.5 kb, which can encode a 59-kDa protein . Although the N-terminal one-fifth of S . cerevisiae TPase (ScCet1p) is missing in CaCet1p, CaCet1p shares significant sequence similarity with ScCet1p over the entire region of the protein; the recombinant CaCet1p, which was expressed as a fusion protein with glutathione S-transferase (GST), displayed TPase activity in vitro . CaCET1 rescued CET1-deficient S . cerevisiae cells when expressed under the control of the ADH1 promoter, whereas the human capping enzyme derivatives that are active for TPase activity but defective in mRNA 5'-guanylyltransferase (GTase) activity did not . Yeast two-hybrid analysis revealed that C . albicans Cet1p can bind to the S . cerevisiae GTase in addition to its own partner, the C . albicans GTase . In contrast, neither the full-length human capping enzyme nor its TPase domain interacted with the yeast GTase . These results indicate that the failure of the human TPase activity to complement an S . cerevisiae cet1delta null mutation is attributable, at least in part, to the inability of the human capping enzyme to associate with the yeast GTase, and that the physical association of GTase and TPase is essential for the function of the capping enzyme in vivo. Mycopathologia, 1998, 141(3), 137 - 42 Monoclonal antibodies directed against extracellular matrix proteins reduce the adherence of Candida albicans to HEp-2 cells; Cotter G et al.; The presence of the extracellular matrix (ECM) proteins collagen types I and IV, laminin and fibronectin on the surface of HEp-2 cells was confirmed by flow cytometry using monoclonal antibodies . Monoclonal antibodies directed against these ECM proteins reduced the adherence of C . albicans ATCC 44990 to HEp-2 cells, the greatest reductions being evident in assays which incorporated anti-collagen type IV monoclonal antibody . The ability of sugaramines to inhibit the adherence of C . albicans to a variety of cell types has been demonstrated previously and the most significant reduction in C . albicans-HEp-2 adherence was in assays which incorporated 0.2M galactosamine . The combination of anti-collagen IV monoclonal antibody and galactosamine reduced the adherence of C . albicans to HEp-2 cells by approximately 70% (p < 0.05). Mycopathologia, 1998, 141(3), 127 - 35 Isolation of a petite mutant from a histidine auxotroph of Candida albicans and its characterization; Roth-Ben Arie Z et al.; Respiration-deficient (petite) mutations have been induced in various yeasts, which are categorized as petite-positive . Candida albicans was classified among the petite-negative yeasts . Since then, a few reports have appeared, describing the isolation of petite mutants in C . albicans . We report in the present study on the isolation of a petite mutant of C . albicans-SAR1 . This mutant was isolated from a histidine auxotroph of C . albicans after mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine, thus our petite mutant carries a double mutation . SAR1 was characterized morphologically, biochemically and ultrastructurally . The results revealed differences from the wild type in respect to morphological, physiological and biochemical characteristics . Electron microscopy showed that the cells of the petite mutant contain only very few mitochondria that looked 'thread like' without any cristae . The significance of the mutation in the virulence of the mutant vs . that of the wild-type is being assessed. J Dent, 1998 Sep, 26(7), 577 - 83 Adherence of Candida albicans to denture-base materials with different surface finishes; Radford DR et al.; OBJECTIVES: To assess the in vitro adherence of Candida albicans to heat-cured hard and soft denture-base materials with varying surface roughness, and to observe the effect of a mixed salivary pellicle on candidal adhesion to these surfaces . METHODS: In vitro adhesion assays on heat-cured acrylic resin (Trevalon), Molloplast B and Novus using the type strain of C . albicans (NCPF 3153A) . Surfaces for the assays were prepared using clinically appropriate rotary instruments . Unstimulated, pooled and clarified whole saliva was used to assess its effect on adhesion . RESULTS: Significantly greater adhesion of C . albicans to rough rather than smooth surfaces was found (P < 0.001), as well as increased adhesion to the machined soft lining materials compared with acrylic . Pre-coating denture-base materials with saliva reduced candidal adhesion on all materials . CONCLUSIONS: Rough surfaces on denture-base materials promote the adhesion of C . albicans in vitro . However, saliva reduces adhesion of C . albicans and thus diminishes the effect of surface roughness and free surface energy differences between materials. Gene, 1998 Sep 18, 218(1-2), 85 - 93 The CARE-2 and rel-2 repetitive elements of Candida albicans contain LTR fragments of a new retrotransposon; Goodwin TJ et al.; CARE-2 and Rel-2 are dispersed, repetitive elements of Candida albicans . Hybridisation experiments suggest that they are present at 10-20 copies per genome and appear on most, if not all, of the chromosomes . A high degree of interstrain variation has been demonstrated for CARE-2, making it of use for strain typing . Until now, however, the nature of the repetitive elements within CARE-2 and Rel-2 was unknown . We show here that CARE-2 and Rel-2 contain long terminal repeat (LTR) fragments of a new retrotransposon . These LTRs, which we designate kappa, are partially responsible for the repetitive nature of CARE-2 and Rel-2 . Complete copies of the kappa elements are present elsewhere in the genome and adjacent to some are sequences characteristic of the internal regions of retrotransposons . An apparently high degree of scrambling of the kappa elements suggests that they may represent a hotspot for mutation and recombination in C . albicans. Mycopathologia, 1998, 141(2), 105 - 9 Activity of hydrolytic enzymes of Candida albicans strains isolated from patients with periodontal and membrane mucosae of oral cavity diseases; Kurnatowska AJ; Fungi are elements of the ontocenosis of the oral cavity and causal factors of inflammatory lesions in its mucous membrane . The objective of the study was to find differences in the activity of hydrolytic enzymes of Candida albicans isolated from patients with diseases of the periodontium and mucous membrane of the oral cavity . Of 235 patients examined, 31 were diagnosed with gingivitis, 38 with glossitis, 28 with leucoplakia, 37 with adult periodontitis, 25 with juvenile periodontitis, 36 stomatitis prothetica and 40 with stomatitis atrophica . In 196 patients (83.4 +/- 2.4%), fungi belonging to Candida species were detected . In the evaluation of Candida albicans strains (146) properties, bioMerieux API ZYM tests containing substrates for the detection of 19 hydrolases were used . All the investigated strains were characterized by the activity of 14 enzymes, i.e . phosphatase alcaline, esterase (C4), esterase lipase (C8), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, alpha galactosidase, beta galactosidase, alpha glucosidase, beta glucosidase, N-acetyl-beta-glucosaminidase, alpha mannosidase and alpha fucosidase . Strains isolated from the oral cavity of patients with disease of periodontium and mucous membrane are characterised by the highest phosphatase acid activity . The greatest enzymatic activity is characteristic of Candida albicans isolated from patients with stomatitis atrophica or stomatitis prothetica, and the lowest in strains from gingivitis or juvenile periodontitis cases . Differences in the activity of hydrolases are statistically significant (p < 0.01) for: esterase (C4), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, beta glucosidase, N-acetyl-beta-glucosaminidase, of fungi isolated from patients with particular clinical diagnoses. Mycopathologia, 1998, 141(2), 59 - 63 Regulation of superoxide dismutase synthesis in Candida albicans; Gunasekaran U et al.; The synthesis of superoxide dismutase {SOD: EC 1.15.1.1} in response to various cultural conditions was examined in Candida albicans, an opportunistic yeast which causes candidiasis in immunosuppressed patients . SOD plays an important role in protecting cells from teh oxidative damage of superoxide radicals . Maximum SOD activity was found after 72 hrs of yeast growth . The optimum pH and temperature for the SOD activity were 7 and 40 degrees C, respectively . The major SOD activity was found in the cytosol fraction and the level of extracellular SOD was very low . The enzyme was stimulated to varying degrees by cholic acid, procaine and tocopherol . On the basis of inhibitor studies and other enzyme properties, the isolated enzyme from C . albicans is identified as a copper and zinc superoxide dismutase. Epidemiol Mikrobiol Imunol, 1998 Aug, 47(3), 87 - 92 {Personal experience with classification of yeast microorganisms . I . The combined biotyping method of Mencl and Otcenásek and typing using the "killer" phenomenon}; Hamal P et al.; Using a group of 150 isolates of Candida albicans, C . glabrata, C . krusei, C . tropicalis, C . parapsilosis a C . kefyr the differentiating capacity of two biotyping systems was tested-the combined method according to Mencl and Otcenasek, and typing using the so-called killer phenomenon . With the combined method comparable results with the original work of the authors were obtained . This applies to the number of biotypes as well as to the ratio of the dominant biotype . As regards the differentiating characteristics of different biotypes the two studies differed fundamentally . As to typing, using the "killer" phenomenon, its practical usefulness was tested, the differentiating capacity of the method was, however, very much influenced by the small number of available killer-positive yeast strains. Ceska Slov Farm, 1998 Jul, 47(4), 186 - 8 {Antimicrobial activity of (N-salicylidene-DL-aspartate- and (N-salicylidene-L-asparaginate)-copper complexes with pyrazole-type ligands}; Sokolik J et al.; By a reaction of salicylaldehyde (Scl) with the corresponding amino acids and by the next complexation reaction of the formed Schiff bases with Cu2+ ions in an aqueous-alcoholic medium, aqua (N-salicylideneaminoalkanoato)copper(II) complex chelates of the composition Cu(Scl-DL-Asp(2-)) (H2O)2, Ip and Cu(Scl-L-Asn(2-)(H2O), In were prepared . The monodiazole complexes with pyrazole IIp and IIn (as monohydrate) as well as with 3,5-dimethylpyrazole IIIp a IIIn were prepared by replacing the molecule of H2O in the parent aquacomplexes with the diazoles under the same reaction conditions . Using a routine dilution micromethod, the antimicrobial activity of the prepared complexes and free diazoles was tested against Staphylococcus aureus, Escherichia coli and Candida albicans . Only a significant antistaphylococcus activity was found (highest for the complex IIn; MIC = 39 micrograms/cm3) . All chelates (Ip,n-IIIp,n) were more effective (MIC = 39-156 micrograms/cm3) than both pyrazole (312 micrograms/cm3)and 3,5-dimethylpyrazole (625 micrograms/cm3) alone . The relationship between the coordination-chemical properties and the biological effects of the complexes studied is discussed. Infect Immun, 1998 Oct, 66(10), 4845 - 50 Mannan-specific immunoglobulin G antibodies in normal human serum accelerate binding of C3 to Candida albicans via the alternative complement pathway; Zhang MX et al.; Candida albicans activates the classical and alternative complement pathways, leading to deposition of opsonic complement fragments on the cell surface . Our previous studies found that antimannan immunoglobulin G (IgG) in normal human serum (NHS) allows C . albicans to initiate the classical pathway . The purpose of this study was to determine whether antimannan IgG also plays a role in initiation of the alternative pathway . Pooled NHS was rendered free of classical pathway activity by chelation of serum Ca2+ with EGTA alone or in combination with immunoaffinity removal of antimannan antibodies . Kinetic analysis revealed a 6-min lag in detection of C3 binding to C . albicans incubated in EGTA-chelated NHS, compared to a 12-min lag in NHS that was both EGTA chelated and mannan absorbed . The 12-min lag was shortened to 6 min by addition of affinity-purified antimannan IgG . The accelerating effect of antimannan IgG on alternative pathway initiation was dose dependent and was reproduced in a complement binding reaction consisting of six purified proteins of the alternative pathway . Both Fab and F(ab')2 fragments of antimannan IgG facilitated alternative pathway initiation in a manner similar to that observed with intact antibody . Immunofluorescence analysis showed that addition of antimannan IgG to EGTA-chelated and mannan-absorbed serum promoted an early deposition of C3 molecules on the yeast cells but had little or no effect on distribution of the cellular sites for C3 activation . Thus, antimannan IgG antibodies play an important regulatory role in interactions between the host complement system and C . albicans. Curr Genet, 1998 Sep, 34(3), 192 - 9 Identification of polymorphic mutant alleles of CaMDR1, a major facilitator of Candida albicans which confers multidrug resistance, and its in vitro transcriptional activation; Gupta V et al.; CaMDR1 (Candida albicans Multi Drug Resistance) encodes a major facilitator whose expression in Saccharomyces cerevisiae confers resistance to several unrelated drugs . We describe here the identification and molecular characterization of seven mutant alleles of CaMDR1 (CaMDR1-1 to 1-7) . The complete sequencing of CaMDR1 alleles revealed several in-frame point mutations leading to a change in amino-acid residues where insertion/replacement of an aspartate residue in a serine-asparagine-aspartate-rich domain was most noteworthy . Interestingly, these alleles showed a distinct drug resistance profile . The expression of CaMDR1, or of its alleles, in C . albicans cells was enhanced by benomyl, methotrexate and several other unrelated drugs, and was more pronounced in at least one of the azole-resistant clinical isolates. Rev Argent Microbiol, 1998 Apr-Jun, 30(2), 100 - 3 {Rapid differentiation and presumptive identificaiton of yeasts using Candida CHROM-agar medium}; Giusiano GE et al.; In order to evaluate faster and cheaper methods for isolation and identification of clinically important yeasts, we used CHROM-agar Candida (CAC), a chromogenic medium, for differentiation and presumptive identification . A total of 546 yeast strains were studied . Strains were previously identified by conventional methods . The colour and texture of Candida albicans, C . tropicalis, C . krusei and Trichosporon beigelii were always constant and particularly distinctive for a reliable identification . C . glabrata, Saccharomyces cerevisiae, C . parapsilopsis colonies also showed constant colour . Because of its contents of cloramphenicol, CAC is also suitable for isolation and observation of mixed yeast species from clinical samples . CAC is a useful medium for reliable differentiation and presumptive identification of clinically important yeasts, particularly form immunocompromised patients. J Immunol, 1998 Sep 15, 161(6), 2684 - 91 HindIII liposomes suppress delayed-type hypersensitivity responses in vivo and induce epidermal IL-10 in vitro; Nishigori C et al.; Considerable evidence suggests that ultraviolet-B (UV-B) radiation suppresses certain immune responses through the induction of cyclobutane pyrimidine dimers in DNA . To determine whether induction of other forms of DNA damage in the skin mimicked the immunosuppressive effects of UV-B radiation, we produced double-strand breaks in the DNA of epidermal cells with HindIII restriction endonuclease encapsulated in liposomes . Application of these liposomes, but not liposomes containing inactive HindIII or an irrelevant endonuclease, to the skin of C3H mice suppressed the induction of delayed-type hypersensitivity responses to Candida albicans and alloantigen and induced IL-10 production in the epidermis . Treatment of the Pam212 murine keratinocyte cell line with these liposomes in vitro induced immunosuppressive activity and IL-10 in culture supernatants . Unlike UV-B irradiation, however, HindIII in liposomes failed to induce suppressor T cell activity in vivo or in vitro . We conclude that double-strand breaks in DNA of epidermal cells can induce immunosuppression and up-regulate the production of immunomodulatory cytokines; however, either DNA damage alone does not account for all the immunosuppressive properties of UV-B irradiation, or cyclobutane pyrimidine dimers differ qualitatively from double-strand breaks in their biologic consequences . These studies raise the possibility that drugs causing DNA damage may induce cytokine dysregulation and immune suppression in addition to cytotoxicity. FEMS Microbiol Lett, 1998 Sep 1, 166(1), 135 - 9 Mitogen-activated protein kinase-defective Candida albicans is avirulent in a novel model of localized murine candidiasis; Guhad FA et al.; Candida albicans strains with a deletion of the mitogen-activated protein kinase CEK1 gene are defective in the yeast to hyphal transition on solid surfaces in vitro . The virulence of a cek1 delta/cek1 delta null mutant strain was compared with its wild-type parent strain (WT) in a novel model of localized candidiasis . The mammary glands of lactating mice (at day 5 postpartum) were infected for 2, 4 and 6 days with 50 microliter suspension containing 1 x 10(5), 1 x 10(6) and 1 x 10(7) blastopores before death . Infected and non-infected control glands were evaluated pathologically . All animals infected with cek1 delta/cek1 delta null mutant strains showed no lesions while 65% of animals infected with the WT strain had severe lesions characterized by widespread heterophilic infiltration, necrosis, and abscess formation . As an additional control, animals infected with the disrupted strain complemented with the WT CEK1, on a replicating plasmid, also showed severe pathological changes similar to the WT strain . These results clearly demonstrate that the CEK1 gene codes for a virulence determinant of C . albicans and that the mouse mastitis model is well suited for the discriminative study of the pathogenicity of different C . albicans strains. Ann Dermatol Venereol, 1997, 124(2), 157 - 8 {Pustular candidiasis in heroin addicts}; Leclech C et al.; INTRODUCTION: Pustular candidiasis in heroin addicts is a rare entity in dermatology . We report a case . CASE REPORT: A 29-year-old female heroin addict developed a painful pustular growth on the scalp . There was no fever . Multiple follicular pustulae measuring 2 to 3 mm were associated with hyperesthesia of the scalp and painful cervical nodes . Biopsy showed acute ostiofolliculitis with a few blastospores and mycelial filaments . Candida albicans was isolated from the pustulae and the buccal cavity . Candida serology was positive (indirect immunofluorescence 1/100, coelectrosyneresis: 4 archs) . Search for other localizations and HIV serology were negative . The last injection of brown heroin had been taken 15 days earlier; lemon had been added . Treatment with flucanazole (400 mg/d) led to improvement within 48 hours . DISCUSSION: Sudden development of pustulae or nodules in pilous zones in a heroin addict should suggest the diagnosis . Outcome depends on early treatment after diagnosis and search for other localizations . Our case presented two particular aspects: ostiofollicular localization of the pustulae and a long delay (15 days) between the (presumably) last injection and the development of the lesion . Folliculitis develops almost exclusively in addicts who use brown heroin . Contamination by Candida albicans results from the lemon used to improve solubility at injection. APMIS, 1998 Jul, 106(7), 736 - 42 Adhesion to denture acrylic surfaces and relative cell-surface hydrophobicity of Candida parapsilosis and Candida albicans; Panagoda GJ et al.; C . parapsilosis is an opportunistic emerging pathogen which together with C . albicans causes diseases in immunocompromised patients . Adhesion of Candida species to various surfaces is an important event in colonization and pathogenesis, and the relative cell-surface hydrophobicity (CSH) of the organism is a contributory physical force involved . Therefore, in vitro adhesion to acrylic surfaces and relative CSH of 24 isolates of C . parapsilosis and 10 isolates of C . albicans were studied . There was no significant difference in relative adhesion of C . parapsilosis isolates and C . albicans, although the former demonstrated a tendency for increased adhesion . There was significant intra-species variation in adhesion among isolates of C . parapsilosis (p=0.0001), but not C . albicans . In general, C . parapsilosis isolates demonstrated a two-fold greater relative CSH than C . albicans (p=0.0003) . When the relative CSH of superficial and systemic isolates of C . parapsilosis were compared, the former showed a significantly higher (49.15%) relative CSH than their systemic counterparts (p<0.01) . A highly significant positive correlation between adhesion and relative CSH of C . parapsilosis (p=0.74, p<0.0001) was also noted . Taken together, these data suggest that the attributes of adhesion and relative CSH of Candida species may contribute differentially in varying disease states of the human host, such as superficial and systemic Candida infections. J Clin Microbiol, 1998 Oct, 36(10), 3007 - 12 Detection of Candida dubliniensis in oropharyngeal samples from human immunodeficiency virus-infected patients in North America by primary CHROMagar candida screening and susceptibility testing of isolates; Kirkpatrick WR et al.; Candida dubliniensis has been associated with oropharyngeal candidiasis in patients infected with human immunodeficiency virus (HIV) . C . dubliniensis isolates may have been improperly characterized as atypical Candida albicans due to the phenotypic similarity between the two species . Prospective screening of oral rinses from 63 HIV-infected patients detected atypical dark green isolates on CHROMagar Candida compared to typical C . albicans isolates, which are light green . Forty-eight atypical isolates and three control strains were characterized by germ tube formation, differential growth at 37, 42, and 45 degreesC, identification by API 20C, fluorescence, chlamydoconidium production, and fingerprinting by Ca3 probe DNA hybridization patterns . All isolates were germ tube positive . Very poor or no growth occurred at 42 degreesC with 22 of 51 isolates . All 22 poorly growing isolates at 42 degreesC and one isolate with growth at 42 degreesC showed weak hybridization of the Ca3 probe with genomic DNA, consistent with C . dubliniensis identification . No C . dubliniensis isolate but only 18 of 28 C . albicans isolates grew at 45 degreesC . Other phenotypic or morphologic tests were less reliable in differentiating C . dubliniensis from C . albicans . Antifungal susceptibility testing showed fluconazole MICs ranging from </=0.125 to 64 microgram/ml . Two isolates were resistant to fluconazole (MIC, 64 microgram/ml) and one strain was dose dependent susceptible (MIC, 16 microgram/ml) . MICs of other azoles, including voriconazole, itraconazole, and SCH 56592, for these isolates were lower . C . dubliniensis was identified in 11 of 63 (17%) serially evaluated patients . Variability in phenotypic characteristics dictates the use of molecular and biochemical techniques to identify C . dubliniensis . This study identifies C . dubliniensis in HIV-infected patients from San Antonio, Tex., and shows that C . dubliniensis is frequently detected in those patients by using a primary CHROMagar screen. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2440 - 2 Microbicidal activity of a new silver-containing polymer, SPI-ARGENT II; Kampf G et al.; The survival of three bacterial species and Candida albicans was studied on SPI-ARGENT II . The immediate recovery from silver-impregnated polymer and control polymer (1 cm2) was approximately 10(6) to 10(7) microorganisms . After incubation (37 degreesC) and neutralization of silver with horse serum (5%), surviving organisms were recovered . The survival of the microorganisms on the polymer was not found to be influenced by the silver implantation. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2446 - 8 Susceptibilities of Candida species isolated from the lower gastrointestinal tracts of high-risk patients to the new semisynthetic echinocandin LY303366 and other antifungal agents; Zhanel GG et al.; Fifty-two percent of stool specimens collected from 1,200 high-risk patients were colonized with yeasts, primarily Candida albicans (53 . 6%) and Candida glabrata (35.7%) . Susceptibilities to all antifungal agents tested, including LY303366, were similar to those reported previously for Candida species isolated from blood. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2437 - 9 Activity of liposomal amphotericin B with prolonged circulation in blood versus those of AmBisome and fungizone against intracellular Candida albicans in murine peritoneal macrophages; van Etten EW et al.; Activity against intracellular Candida albicans was assessed in C . albicans-infected murine peritoneal macrophages exposed to long-circulating pegylated amphotericin B liposomes (PEG-AMB-LIP), AmBisome, or Fungizone . The level of antifungal activity of Fungizone is much higher than that of AmBisome or PEG-AMB-LIP, while PEG-AMB-LIP and AmBisome show equivalent activity levels . Previous exposure of uninfected macrophages to PEG-AMB-LIP or AmBisome is advantageous for intracellular antifungal activity. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2434 - 6 Control of Candida albicans murine vaginitis by topical administration of polycarbophil-econazole complex; Ghelardi E et al.; The complexation of econazole with the mucoadhesive polycarbophil was found to significantly improve the therapeutic benefit of the drug in the topical treatment of experimental vaginal candidiasis in mice, while no difference in the antimycotic activity exerted by econazole and polycarbophil-econazole could be detected in vitro. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2431 - 3 Superior efficacy of liposomal amphotericin B with prolonged circulation in blood in the treatment of severe candidiasis in leukopenic mice; van Etten EW et al.; In leukopenic mice with severe systemic candidiasis, single-dose treatment (5 mg of amphotericin B {AMB}/kg of body weight) with long-circulating polyethylene glycol-coated AMB liposomes (PEG-AMB-LIP) resulted in zero mortality and a significant reduction in the number of viable Candida albicans in the kidney, whereas 70% mortality was seen in mice treated with five daily doses of AmBisome (5 mg of AMB/kg . day) . When the first of five daily doses of AmBisome was combined with a single low dose of Fungizone (0.1 mg of AMB/kg), the efficacy was equal to that of PEG-AMB-LIP. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2405 - 9 Pharmacokinetics of pentoxifylline and its metabolites in healthy mice and in mice infected with Candida albicans; Miller K et al.; Pentoxifylline has immunomodulatory properties and has been shown to decrease organ damage and improve survival in animals with gram-negative sepsis or endotoxemia . This effect is mediated by a reduction in endotoxin-induced production of tumor necrosis factor alpha (TNF-alpha) by the host . In earlier studies, we observed an unexpected increase in mortality in mice infected with Candida albicans that were given pentoxifylline even though concentrations of TNF-alpha in serum were not affected . The current study was designed to determine whether the pharmacokinetics of pentoxifylline and its metabolites were altered in C . albicans-infected mice and, if so, whether these changes could have contributed to the increased mortality . Noninfected mice and mice infected with C . albicans were treated with pentoxifylline (60 mg/kg of body weight) intraperitoneally every 8 h . Serum was collected from animals after one (day 0), four (day 1), or seven (day 2) injections of pentoxifylline or saline (controls) . The first dose was administered 6 h after C . albicans infection . Serum was pooled . Concentrations of pentoxifylline and metabolites I, IV, and V were determined by capillary gas chromatography . Renal function and hepatic profiles were assessed . Pharmacokinetic parameters (maximum concentration of pentoxifylline in serum, half-life, and area under the concentration-time curve from 0 h to infinity {AUC(0)-infinity}) for all noninfected mice were similar and did not differ from those for day 0-infected mice . For day 1-infected mice, values of these three pharmacokinetic parameters for pentoxifylline and metabolite I were increased two- to fourfold over values for noninfected and day 0-infected mice . For metabolites IV and V, the AUC(0)-infinity was increased approximately eightfold over control values . In addition, day 1-infected mice demonstrated evidence of renal and hepatic dysfunction . In summary, C . albicans infection produced marked changes in the pharmacokinetics of pentoxifylline and its metabolites in the mice . The high concentrations of pentoxifylline and its metabolites in serum attained in infected mice may have contributed to the increased mortality of mice with systemic candidiasis. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2342 - 6 Synergy of nitric oxide and azoles against Candida species in vitro; McElhaney-Feser GE et al.; The candidacidal activity of nitric oxide (NO) as delivered by a class of compounds termed diazeniumdiolates has been investigated . Diazeniumdiolates are stable agents capable of releasing NO in a biologically usable form at a predicted rate, and three such compounds were examined for activity . One compound, (Z)-1-{N-(2-aminoethyl)-N-(2-ammonioethyl)amino}diazen-1- ium-1, 2-diolate (DETA-NO), proved to be most suitable for examining NO activity due to its relatively long half-life (20 h) and because of limited candidacidal activity of the uncomplexed DETA nucleophile . DETA-NO was active against six species of Candida for which the MICs necessary to inhibit 50% growth (MIC50s) ranged from 0.25 to 1.0 mg/ml . C . parapsilosis and C . krusei were the most susceptible to the compound . In addition to a determination of NO effects alone, the complex was utilized to investigate the synergistic potential of released NO in combination with ketoconazole, fluconazole, and miconazole . Activity was investigated in vitro against representative strains of Candida albicans, C . krusei, C . parapsilosis, C . tropicalis, C . glabrata, and C . dubliniensis . Determination of MIC50, MIC80 and MICs indicated that DETA-NO inhibits all strains tested, with strains of C . parapsilosis and C . krusei being consistently the most sensitive . The combination of DETA-NO with each azole was synergistic against all strains tested as measured by fractional inhibitory concentration indices that ranged from 0.1222 to 0.4583 . The data suggest that DETA-NO or compounds with similar properties may be useful in the development of new therapeutic strategies for treatment of Candida infections. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2279 - 83 Identification of elongation factor 2 as the essential protein targeted by sordarins in Candida albicans; Dominguez JM et al.; The target for sordarins in Candida albicans has been elucidated . Kinetic experiments of sordarin inhibition as well as displacement experiments showed that the formation of a sordarin-target complex follows a reversible mechanism . Binding of tritiated drug to the target is enhanced in the presence of ribosomes . Isolation of the target by classical protein purification methods has allowed us to identify it as elongation factor 2 . This is in agreement with the nature of sordarin derivatives as specific inhibitors of the elongation cycle within protein synthesis in yeasts. Antimicrob Agents Chemother, 1998 Sep, 42(9), 2197 - 205 Molecular mode of action of the antifungal beta-amino acid BAY 10-8888; Ziegelbauer K et al.; BAY 10-8888 is a cyclic beta-amino acid that is related to cispentacin and that has antifungal activity . Candida albicans cells accumulated BAY 10-8888 intracellularly to a concentration about 200 that in the medium when grown in media with a variety of nitrogen sources . In complex growth medium, BAY 10-8888 transport activity was markedly reduced and was paralleled by a decrease in its antifungal activity . Uptake of BAY 10-8888 was mediated by an H+-coupled amino acid transporter with specificity for branched-chain amino acids (isoleucine, leucine, and valine) and showed a KT (Michaelis constant of the transport reaction) of 0.95 mM and a Vmax of 18.9 nmol x min-1 x 10(7) cells-1 . Similar to the transport of natural amino acids in Saccharomyces cerevisiae, the transport of BAY 10-8888 into the cell was unidirectional . Efflux occurred by diffusion and was not carrier mediated . Inside the cell BAY 10-8888 inhibited specifically isoleucyl-tRNA synthetase, resulting in inhibition of protein synthesis and cell growth . Intracellular isoleucine reversed BAY 10-8888-induced growth inhibition . BAY 10-8888 was not incorporated into proteins . BAY 10-8888 inhibited isoleucyl-tRNA synthetase with the same concentration dependency as protein biosynthesis in intact cells assuming 200-fold accumulation. Immunopharmacol Immunotoxicol, 1998 Aug, 20(3), 421 - 31 Protective effect of oral administration of a traditional medicine, Juzen-Taiho-To, and its components on lethal Candida albicans infection in immunosuppressed mice; Abe S et al.; Protective effects of a kampo medicine, Juzen-taiho-to (TJ-48) and its herbal components against experimental candidiasis in cyclophosphamide-induced immunosuppressive mice were investigated . ICR mice were immunosuppressed by intraperitoneal treatment with cyclophosphamide (day-4) and were orally given TJ-48 or one of its 10 herbal components for 4 consecutive days (day-4--1) . They were then challenged intravenously with a lethal dose of Candida albicans (day 0) . An oral dose of 1 g/kg/day of TJ-48 prolonged their life span . A similar protective effect was obtained by treatment with its component drugs Ginseng radix, Glycyrrhizae radix, Atractylodis lancea rhizoma or Cnidii rhizoma . These herbal components were suggested to have a main role in the protective effect of Juzen-taiho-to against Candida infection. Bol Asoc Med P R, 1998 Jan-Mar, 90(1-3), 21 - 6 Anti-fungal and cytokine producing activities of CD8 + T lymphocytes from HIV-1 infected individuals; Colon MD et al.; Lymphokine activated killer (LAK) cells are capable of killing not only malignant cells but also hyphal form of Candida albicans in vitro . When peripheral blood mononuclear cells (PBMC) from normal healthy donors were cultured for 72-96 hrs with 1,500 international unit (IU)/ml interleukin-2 (IL-2), marked LAK activity was induced . However, even prior to IL-2 activation, PBMC isolated from some normal subjects and those from almost all individuals who are infected by human immunodeficiency virus type 1 (HIV-1) exhibited significant levels of anti-fungal activity . Such pre-activation ("in situ") antifungal activity of PBMC decreased during the initial 48 hrs of IL-2 activation . PBMC from HIV-1 seropositive subjects showed higher levels of "in situ" anti-fungal activity than normal PBMC did . After a decline of "in situ" activity during the initial 48 hours, LAK activity gradually increased and reached near maximal levels by day 4 and remained more or less constant until day 6 . No significant difference was observed between the LAK activity of normal and HIV-1(+) PBMCs on days 4-6 . In IL-2 activated normal and HIV-1(+) PBMC cultures, both CD4 and CD8 T cells produced IL-2, INF-gamma as well as TNF-alpha . Production of IL-2 by both CD4 and CD8 T cells was suppressed in HIV-1(+) PBMC cultures, but no significant suppression of INF-gamma production was noted . Meanwhile, TNF-alpha production by CD4 was very much suppressed but no significant changes in TNF-alpha production by CD8 T cells was noted in HIV-1(+) PBMC cultures. Scand J Infect Dis, 1998, 30(2), 137 - 42 Outbreak of Candida albicans fungaemia in a neonatal intensive care unit; Huang YC et al.; During a 4-month period, 9 premature infants hospitalized in a neonatal intensive care unit (NICU) developed Candida albicans fungaemia . All 9 infants received antifungal agents . Fluconazole was administered in 7 patients and successfully eradicated this organism in 6 with no adverse effects . For epidemiological investigation, 64 environmental specimens and hand-washings of all 54 staff members involved in the NICU were examined for the presence of this organism . No C . albicans could be identified from environmental sources, while the hand-washing of 1 nurse was C . albicans-positive . Two genotyping methods, including electrophoretic karyotyping using contour-clamped homogeneous electric field gel electrophoresis and polymerase chain reaction-based direct sequencing of rRNA gene, were used in the analysis of the isolates recovered from blood cultures of the infants, the hand-washing of the nurse and 7 control isolates . Both methods yielded comparable results and revealed that all 13 isolates from infected infants as well as the isolate from hand washing of the nurse were of the same genotype while the control isolates were distinct . These results suggest that the outbreak of C . albicans fungaemia was caused by a particular strain and possibly via cross-infection . In addition, we showed that fluconazole seemed to be safe and effective in treating C . albicans fungaemia in neonates, although the data were limited. J Infect Dis, 1998 Sep, 178(3), 792 - 802 Early signal transduction induced by Candida albicans in macrophages through shedding of a glycolipid; Jouault T et al.; Cell wall beta-1,2-oligomannosides are involved in Candida albicans binding to macrophages and in their stimulation to produce cytokines . The nature of signaling events occurring during initial interaction of macrophage J774 cell line and C . albicans, together with the nature of molecules containing beta-1,2-oligomannosides released by the yeasts, was examined . Cocultivation led to a herbimycin A-sensitive production of tumor necrosis factor-alpha . Immunofluorescence and Western blotting confirmed tyrosine phosphorylation and revealed an accumulation of 90- to 120-kDa phosphoproteins . Antibodies specific for beta-1,2-oligomannosides showed that these epitopes were shed at an early stage from the yeasts to the macrophage membrane, in association with a glycolipid previously described as C . albicans phospholipomannan . Incubation of macrophages with purified phospholipomannan alone led to a signal transduction pathway identical to that observed with living yeasts . All of these results demonstrate that C . albicans phospholipomannan shedding is involved in C . albicans-macrophage interaction through beta-1,2-oligomannosides. Eur J Pediatr, 1998 Aug, 157(8), 661 - 2 Pharmacokinetics of oral fluconazole in premature infants; Wenzl TG et al.; Systemic infections with Candida albicans in neonates are a frequent and well recognized problem . The therapeutic gold standard in this situation is the combined intravenous antimycotic treatment with amphotericin B and flucytosine . Potential adverse effects of this regimen have encouraged the search for desirable alternatives . We report on the use of oral fluconazole in neonates with Candida albicans septicaemia . Three premature infants were treated with four courses of therapy . Pharmacokinetic studies were performed during each course . At oral doses of 4.5-6 mg/kg once a day, serum levels of fluconazole were within the therapeutic range during the entire dosage interval . Follow up showed microbiological and clinical cure in all patients with no side-effects . In one patient a dosage of 4 mg/kg per day lead to a microbiological relapse with sub-therapeutic serum levels . CONCLUSIONS: Oral fluconazole seems to be a safe and effective treatment for Candida albicans septicaemia even in premature infants. Eur Respir J, 1998 Aug, 12(2), 502 - 4 Eosinophilic lung disease associated with Candida albicans allergy; Pacheco A et al.; A significant number of cases of chronic eosinophilic pneumonia remain idiopathic in spite of a comprehensive search of associated causes . This study reports a patient with a classical clinical presentation of chronic eosinophilic pneumonia and peripheral blood eosinophilia in whom selective sensitization to Candida albicans was demonstrated . This yeast was present in the bronchoalveolar lavage culture and specific serum immunoglobulin (Ig)E and IgG against C . albicans were found in the patient's serum . Levels of these specific immunoglobulins diminished with corticosteroid treatment and increased coinciding with a new outbreak of symptoms after lowering the dosage of corticosteroids . To the author's knowledge, this is the first case described of chronic eosinophilic pneumonia associated with sensitization to C . albicans . Evaluation of allergy to C . albicans should be performed in chronic eosinophilic pneumonia before labelling cases as idiopathic. J Antibiot (Tokyo), 1998 Jul, 51(7), 665 - 75 Characterization and optimization of in vitro assay conditions for (1,3)beta-glucan synthase activity from Aspergillus fumigatus and Candida albicans for enzyme inhibition screening; Wood RL et al.; (1,3)Beta-D-glucan synthase (E.C.2.4.1.34 . UDP-glucose: 1,3-beta-D-glucan 3-beta-glucosyl transferase) catalyzes the polymerization of glucose ({1-3}-beta-linkages) using UDP-glucose as substrate . We have determined optimal in vitro conditions for the assay of (1,3)beta-glucan synthase activity from Aspergillus fumigatus and Candida albicans . These included lysis of cells in the following for C . albicans, 100 mM HEPES, pH 8.0, 10 microM guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS), 2 mM ethylenediaminetetraacetic acid (EDTA), disodium salt, 5 mM NaF, 250 mM sucrose, and 10 mM NaH2PO4; and for A . fumigatus, 50 mM HEPES, 10mM EDTA, 750 mM sucrose, 10 mM NaH2PO4, 100 mM cellobiose and 50 microM GTPgammaS . Resulting low-speed supernatants were used as enzyme sources to determine the optimal in vitro assay conditions . We have characterized the resulting enzyme activities and tested the optimized assays with known (1,3)beta-glucan synthase inhibitors including cilofungin, papulacandin, aculeacin A, and echinocandin B . We have used both optimized assays to screen > 1000 extracts of marine macroorganisms and, using bioassay-guided purification, have identified (1,3)beta-glucan synthase inhibitors. Mycopathologia, 1998, 141(1), 1 - 6 Factors influencing the expression in vitro of Candida albicans stress mannoproteins reactive with salivary secretory IgA; Vidotto V et al.; We have examined the influence of subinhibitory concentrations of several antifungals, the different glucose and ammonium sulphate concentrations in the culture medium as well as the strain variability on the expression in vitro of stress mannoproteins reactive with salivary sIgA in C . albicans and other Candida spp isolates . Irrespective of the conditions used, no reactivity with salivary sIgA was observed in yeast cells grown at 25 degrees C . However, when grown at 37 degrees C, all of the 10 C . albicans strains, but only 9 out of 28 non-C . albicans isolates studied showed reactivity with salivary sIgA . Cells grown at 37 degrees C in medium containing maximum concentrations of glucose and ammonium sulphate expressed the antigens reactive with sIgA during longer periods of time than the cells grown in medium with minimal concentrations of the same compounds . The regulatory role showed by the concentration of glucose and ammonium sulphate on the antigenic expression was subordinated, nevertheless, to the most important factor, the temperature of incubation . Only isolates showing low susceptibility expressed the antigens reactive with sIgA under the influence of subinhibitory concentration of antifungals . However, induced resistance to one of the antifungals tested (5 fluorocytosine) allowed the antigenic expression at elevated subinhibitory concentrations even in previous susceptible strains . In conclusion, in addition to the temperature, factors such as characteristics of the strain, the concentration of glucose and ammonium sulphate in the culture medium and the resistance to antifungals played a role on the expression of C . albicans antigens reactive with sIgA, which could be of clinical relevance in the course of infection. Mol Microbiol, 1998 Aug, 29(3), 753 - 62 Deletion analysis of yeast Sec65p reveals a central domain that is sufficient for function in vivo; Regnacq M et al.; The Saccharomyces cerevisiae SEC65 gene encodes a 32 kDa subunit of yeast signal recognition particle that is homologous to human SRP19 . Sequence comparisons suggest that the yeast protein comprises three distinct domains . The central domain (residues 98-171) exhibits substantial sequence similarity to the 144 residue SRP19 . In contrast, the N-terminal and C-terminal domains (residues 1-97 and 172-273 respectively) share no similarity to SRP19, with the exception of a cluster of positively charged residues at the extreme C-terminus of both proteins . Here, we report the cloning of a Sec65p homologue from the yeast Candida albicans that shares the same extended domain structure as its S . cerevisiae counterpart . This conservation of sequence is reflected at the functional level, as the C . albicans gene can complement the conditional lethal sec65-1 mutation in S . cerevisiae . In order to examine the role of the N- and C- terminal domains in Sec65p function, we have engineered truncation mutants of S . cerevisiae SEC65 and tested these for complementing activity in vivo and for SRP integrity in vitro . These studies indicate that a minimal Sec65p comprising residues 76-209, which includes the entire central SRP19-like domain, is sufficient for SRP function in yeast. Ann Allergy Asthma Immunol, 1998 Aug, 81(2), 165 - 9 Recurrent vaginal candidiasis and allergic rhinitis: a common association; Moraes PS; BACKGROUND: In a recent series of studies, it has been shown that hypersensitivity is an important factor in recurrent vaginal candidiasis and several referring gynecologists have been asking for our support in the evaluation of women with regard to the possibility of local vaginal hypersensitivity . This study was instituted because, since the first anamnesis, a high incidence of perennial allergic rhinitis, as well as family allergies have been observed in these patients . OBJECTIVE: To study the association between recurrent vaginal candidiasis and perennial allergic rhinitis, and to explore the allergic characteristics of the patients and of the disease . METHODS: For 28 months, we have prospectively studied 95 patients with recurrent vaginal candidiasis referred by gynecologists to a private allergy practice . All of them were unresponsive to all other modalities of therapy, and had no diabetes or immunodeficiency diseases . As a control group, we studied 100 women, who came to the allergy office for other reasons, and had no recurrent vaginal candidiasis . All of these 195 women were submitted to a standard allergy medical history, a complete physical examination, and immediate skin tests with a standard battery of inhalant allergens and Candida albicans . The incidence of allergic diseases in the two groups was compared . RESULTS: Sixty-four patients with recurrent vaginal candidiasis also had allergic rhinitis (71%), while in the control group the incidence of this pathology was 42% . The difference was considered statistically significant, according to the software Epi info (P < .0001) . The study (1) did not show an association between recurrent vaginal candidiasis and asthma and (2) indicated that patients with recurrent vaginal candidiasis have a high incidence of skin tests positive to inhalant allergens (50%), to Candida albicans (55%), and a high incidence of family history of allergies (73%) . In control women, the incidence of skin test positive to inhalant allergens was 72% and to Candida albicans, 10% . There was a family history of allergies in 61% of women . CONCLUSIONS: The data demonstrate that recurrent vaginal candidiasis is statistically associated with perennial allergic rhinitis and that many of these women with recurrent vaginal candidiasis tend to be atopic. J Biol Chem, 1998 Sep 4, 273(36), 23376 - 80 A functional soluble form of the murine mannose receptor is produced by macrophages in vitro and is present in mouse serum; Martinez-Pomares L et al.; A soluble form of the mannose receptor (sMR) has been found in conditioned medium of primary macrophages in vitro and in mouse serum . sMR was released as a single species, had a smaller size than the cell-associated form, and accumulated in macrophage-conditioned medium, in a cytokine-regulated manner, to levels comparable with those found for cell-associated mannose receptor . Pulse-chase experiments showed that sMR production in culture occurred by constitutive cleavage of pre-existing full-length protein . A binding assay was developed to determine the sugar specificity of sMR and its ability to interact with pathogens and particulate antigens (i.e . Candida albicans and zymosan) . Protease inhibitor studies suggested that sMR was produced by cleavage of an intact mannose receptor by a matrix metalloprotease or ADAM metalloprotease . A role for sMR in the immune response is proposed based on its binding properties, regulation by cytokines, and the previous discovery of putative ligands for the cysteine-rich domain of the mannose receptor in lymph nodes and spleen. Mol Microbiol, 1998 Jul, 29(2), 605 - 15 Differential expression of secreted aspartyl proteinases in a model of human oral candidosis and in patient samples from the oral cavity; Schaller M et al.; Candida albicans, an opportunistic pathogen in humans, secretes secretory aspartyl proteinases (Saps), which have been correlated with virulence . We examined the temporal regulation of the mRNA expression of seven known members of the SAP gene family by reverse transcription polymerase chain reaction (RT-PCR) in (i) an in vitro model of oral candidosis based on reconstituted human epithelium (RHE); and (ii) clinical samples from patients with oral candidosis . SAP1 and SAP3 transcripts were first detected 42 h after inoculation of RHE, while at the same time, slight morphological alterations in the epithelium were documented by light microscopy . SAP6 expression occurred 6 h later concomitantly with germ tube formation of some infecting Candida cells and severe lesions of the epithelial tissue . SAP2 and SAP8 RT-PCR products were first detected 60 h after infection, while SAP4 and SAP5 transcripts were never discovered . Thus, a temporal progression of SAP expression in the order SAP1 and SAP3 > SAP6 > SAP2 and SAP8 was observed at the same time as increasing RHE damage occurred . At the protein level, Sap antigen was found within the C . albicans yeast cells and the epithelial cells by immunoelectron microscopy using an anti-Sap murine monoclonal antibody directed against the gene products Sap1-3 . Expression of SAP1-3 and 6 was also detected by RT-PCR in samples from patients suffering from oral candidosis . Our results suggest that the pathogenesis of experimental and clinical oral candidosis is associated with the differential and temporal regulation of SAP gene expression. Clin Exp Allergy, 1998 Jul, 28(7), 808 - 16 Requirement of CD28-CD86 costimulation for allergen-specific T cell proliferation and cytokine expression; Van Neerven RJ et al.; BACKGROUND: Allergen-specific T lymphocytes biased to the production of type 2 cytokines play an important role in the pathophysiology of atopic disease . It is not known whether optimal activation of these T cells requires costimulation via interaction of B7 (CD86/CD80) with CD28 . METHODS: Peripheral blood mononuclear cells (PBMC), isolated from 10 house dust mite Dermatophagoides pteronyssinus (Der p)-allergic asthma patients and 10 non-allergic control individuals, were stimulated with house dust mite (Der p) and the control antigens Candida albicans (CA) and tetanus toxoid (TT) . The role of costimulation in activation for proliferation and cytokine mRNA production of peripheral blood T cells was studied by blocking CD28, CTLA-4, and their ligands CD80 (B7-1) and CD86 (B7-2) . RESULTS: The proliferation and the production of type 1 and 2 cytokine mRNA by T cells in response to Der p as well as the control antigens TT and CA was inhibited by simultaneously masking CD80 and CD86 using CTLA4-Ig, a soluble form of CTLA-4 . Notably, Der p-specific proliferation of T cells from Der p-allergic asthma patients and non-allergic controls were inhibited equally well . Additional experiments with MoAbs revealed that activation of these T cells was optimally inhibited by blocking the interaction of CD28 with CD86 . CONCLUSION: In vitro responses of allergen- and antigen-specific T cells of allergic patients and non-allergic control persons are equally dependent on costimulation via the CD28-CD86 pathway, suggesting that inhibition of this pathway may prevent complete activation of allergen-specific T cells in allergic individuals in vivo. Microbiology, 1998 Aug, 144 ( Pt 8), 2311 - 21 Disruption studies of a Candida albicans gene, ELF1: a member of the ATP-binding cassette family; Sturtevant J et al.; A 3.6 kb gene (ELF1) with homology to the ATP-binding cassette (ABC) gene family has been isolated from genomic libraries of Candida albicans . Members of this gene family include both membrane transport proteins which confer a drug-resistance phenotype, and proteins whose functions are associated with protein translation . ELF1 (Elongation Like Factor) showed greatest homology with a Saccharomyces cerevisiae ORF (YPL226W), whose function is unknown, and lower homology with fungal elongation factor 3 (EF-3) genes . In comparison, homology with a gene conferring a drug-resistant phenotype (CDR1) was low . To understand the function of ELF1 in C . albicans, gene-knockout experiments were conducted using the hisG-URA3-hisG disruption cassette . Both single-copy (heterozygote) and double-disrupted strains in ELF1 were isolated . Phenotypically, the disrupted strains grew more slowly than wild-type and produced a mixture of large, irregular cells and apparently normal cells. Microbiology, 1998 Aug, 144 ( Pt 8), 2299 - 310 Overexpression of Candida albicans secretory aspartyl proteinase 2 and its expression in Saccharomyces cerevisiae do not augment virulence in mice; Dubois N et al.; To elucidate the implications of secreted aspartyl proteinase (Sap)2p in the pathogenesis of Candida infections, the SAP2 gene was expressed in Saccharomyces cerevisiae and overexpressed in Candida albicans . The coding region of SAP2, including its signal sequence and propeptide, was amplified by PCR and cloned downstream of the S . cerevisiae or C . albicans ADH1 promoter . Plasmid expression of SAP2 in S . cerevisiae showed that the signal peptide was functional . Integrative transformation of S . cerevisiae and C . albicans was accomplished by homologous recombination within the URA3 locus for S . cerevisiae and the SAP2 locus for C . albicans . Negative control transformants carried plasmids either without the SAP2 insert or with mutated sap2 . S . cerevisiae and C . albicans transformants showed similar growth rates to their parental strains or negative controls, when grown in medium containing amino acids . However, in medium with BSA as sole nitrogen source, constitutive expression of SAP2 enabled S . cerevisiae to grow and increased the growth rate of C . albicans . In both media, only S . cerevisiae transformants harbouring SAP2 secreted the enzyme, as confirmed by proteinase activity assays and immunoblotting . When C . albicans was grown in amino acids medium, the enzyme was detected exclusively in transformants constitutively expressing SAP2 . However, in BSA medium these strains secreted enzyme earlier and secreted higher amounts of enzyme and total proteinase activity . In pathogenicity studies in intact mice, expression of Sap2p as a sole putative virulence factor did not cause S . cerevisiae to become virulent and constitutive overexpression of SAP2 did not augment virulence of C . albicans in experimental oral or systemic infection. Microbiology, 1998 Aug, 144 ( Pt 8), 2291 - 8 Evaluation of the intranasal challenge route in mice as a mucosal model for Candida albicans infection; Londono P et al.; The intranasal route was used to study Candida albicans infections in mice . Mice from two different inbred strains were challenged intranasally with C . albicans and the level of local and systemic colonization was monitored . DBA/2 mice were highly susceptible to challenge and viable C . albicans disseminated from the lungs to deeper tissues, including kidneys, liver and spleen within 48 h . In contrast, in BALB/c mice challenged in the same manner, C . albicans were retained within the lungs and cleared . Local and systemic anti-C . albicans immune responses were investigated . BALB/c mice exhibited higher titres of serum and mucosal anti-C . albicans IgA than DBA/2 mice . Splenocytes from BALB/c mice, but not from DBA/2 mice, produced detectable levels of interleukin-4 and -5 following stimulation with C . albicans antigens . Both DBA/2- and BALB/c-derived splenocytes produced interferon-gamma and interleukin-10 in response to similar stimulation . In conclusion, the intranasal route provided a simple, non-invasive murine model for investigating C . albicans infection through mucosal surfaces. FEMS Immunol Med Microbiol, 1998 Jul, 21(3), 223 - 30 Soluble mannan and beta-glucan inhibit the uptake of Malassezia furfur by human monocytic cell line, THP-1; Suzuki T et al.; The uptake of live and heat-killed Malassezia furfur HIC 3321, HIC 3343 and Candida albicans ATCC 10231 by human monocytic cell line, THP-1, was examined . THP-1 was differentiated by PMA for 7 days before use . The uptake of these yeasts by THP-1 was increased in a concentration-dependent manner of yeasts, and the uptake reached plateau level at the E/T (yeast/THP-1) ratio 5 . In addition, a higher percentage of heat-killed cells than live cells was taken in THP-1 . Yeast mannan and beta-1,3-glucan, random coiled conformer, inhibited the uptake of live and heat-killed M . furfur by THP-1, though dextran T-250, that is alpha-glucan, and schizophyllan (SPG), triple helix conformer of beta-glucan, did not . Interestingly, mannan inhibited the uptake of both types, live and heat-killed, of C . albicans, however, laminaran inhibited the uptake of heat-killed C . albicans alone . Opsonization of these yeasts with normal human serum enhanced the uptake of yeasts, although opsonization with heat-inactivated serum, the treatment at 56 degrees C for 30 min, did not enhance . These results suggested that live and heat-killed M . furfur was recognized by THP-1 through mannose receptor, beta-glucan receptor and complement receptor type 3 via the activation of alternative pathway of complement. Mycoses, 1998, 41 Suppl 1, 71 - 7 {Differentiation and characterization of yeasts pathogenic for humans (Candida albicans, Exophiala dermatitidis) and algae pathogenic for animals (Prototheca spp.) using Fourier transform infrared spectroscopy (FTIR) in comparison with conventional methods}; Schmalreck AF et al.; Due to the Fourier-Transform Infrared Spectroscopy (FT-IR) of strain specific traits demonstrated to be a suitable and efficient method for diagnostic and epidemiological determinations for the yeasts Candida albicans, Exophiala dermatitidis and the chlorophylless algae of the genus Prototheca . FT-IR leads in a rapid and economical way to reproducible results according to the spectral differences of intact cells (IR-fingerprints) . Different genera, species and sub-species respectively, different strains can be recognized and grouped into different clusters and subclusters . The FT-IR analysis of Candida albicans isolates (n = 150) of 22 newborns-at-risk of an intensive care unit showed, that 86% of the children were colonised with several (2-4) different strains in the oral cavities and faeces . Stationary cross-infections could definitely be determined . Exophiala dermatitidis isolates (n = 31), mostly isolated repetitively within a period of 3 years from sputa of patients suffering from cystic fibrosis could be characterized and grouped patient-specifically over the total sampling period . Of 6 from 8 patients (75%) their individual strains remain the same and could be tracked over the three years . Cross-infections during the stationary treatment could be clearly identified by FT-IR . The Prototheca isolate (n = 43) from live-stock and farm environment showed clear distinguishable clusters differentiating the species P . wickerhamii, P . zopfii and P . stagnora . In addition, the biotypes of P . zopfii could be distinguished, especially the subclusters of variants II and III . It could be demonstrated, that FT-IR is suitable for the routine identification and differentiation of yeasts and algae . However, in spite of the gain of knowledge by using FT-IR for the characterization of microorganisms, the conventional phenotyping and/or genetic analysis of yeast or algae strains cannot be replaced completely . For a final taxonomic classification a combination of conventional methods on FT-IR together with more sophisticated molecular genetic procedures is necessary. Mycoses, 1998, 41 Suppl 1, 47 - 50 {What functions do six different genes for secretory proteinases have in Candida albicans?}; Hube B et al.; Secreted Aspartate Proteinases (Sap) are among those factors of the human pathogen Candida albicans, which promote infections in the immunocompromised host . Sap isoenzymes are encoded by at least nine different genes (SAP1-9), which are differentially regulated in vitro . RT-PCR analysis during experimental infections and from patient samples confirmed the expression of SAP genes in vivo . However, while Sap2 is the dominant isoenzyme under culture conditions, other SAP genes are also expressed during infections . In order to investigate the role of single isoenzymes during the pathogenesis of candidosis, mutants were produced which harbour deletions in SAP1, SAP2, SAP3 and SAP4-6 . Although only SAP2 and SAP4-6 mutants showed a strong reduction of proteolytic activity in vitro, all SAP mutants were significantly attenuated in systemic infections . In addition, SAP2, SAP3 and SAP4-6 mutants were clearly more sensitive to neutrophilic leucocytes compared to the wild type SC5314 . These investigations show that several proteinase isoenzymes are likely to be involved in the pathogenesis of candidosis. Mycoses, 1998, 41 Suppl 1, 39 - 46 {Qualitative and quantitative studies of autofluorescence in fungi}; Graf B et al.; Fluorescence microscopy is an important method in mycology . It is a common procedure used in immunology or histology and more recently in modern techniques of molecular biology like in-situ hybridization . Since several molds and yeasts show autofluorescence, an interference of this phenomenon with the detection method cannot be excluded . Therefore, we studied autofluorescence in fungi in more detail, in particular with respect to the dependence of this phenomenon from growth conditions, fixing method or mounting medium used . Here we show that moulds cultivated in a liquid medium are strongly autofluorescent which could be considerably reduced by repetitive washing . In moulds, we did not find important differences in autofluorescence levels with the three fixing methods under study . However, this finding cannot be generalized . Thus, in the yeast Candida albicans we found the autofluorescence pattern being largely dependent from the fixing method and the excitation wave length, respectively . In particular, with green excitation we could show that aceton fixation resulted in strong fluorescence of individual cells within a vast population of cells showing little or no autofluorescence . In addition, we could demonstrate that mounting media are able to strongly modify autofluorescence in fungi . Using digital image acquisition with a cooled CCD camera we were able to quantify the influence of different mounting media on fluorescence intensities of Aspergillus fumigatus. Bone Marrow Transplant, 1998 Jul, 22 Suppl 1, S48 - 51 Detection of cell surface determinants for anti-Leu M3 (CD14), MY9 (CD33) and MY4 (CD14) and phagocytic function of cord blood monocytes in the course of gestational age; Gengenbacher D et al.; To investigate immunological functions of cord blood cells, cord blood monocytes of 84 term and preterm newborn infants were examined in presenting cell surface determinants for the monoclonal antibodies anti-Leu M3 (CD14), My9 (CD33) and My4 (CD14) using an alkaline-phosphatase-anti-alkaline-phosphatase staining method . We also tested the phagocytic function of these cells by incorporation of Candida albicans in a 60 min incubation assay . Mean values for antigen expression ranged from 60 to 70%, whereby My4 showed a statistically significant higher level compared to Anti-Leu M3 and My9 (P = 0.05) . Changes of antigen expression were statistically significant by the time of maturation . Statistically significant differences between male and female newborns were observed in 26-33 and 36-37 weeks of gestation . Considering the modus of delivery, no significantly higher amount of antigen-presenting monocytes were found in newborns of spontaneous birth compared to cesarean sections . However, no statistically significant differences in phagocytosis were observed between term and preterm newborn infants, mature newborns showed higher levels of both antigen expressing and phagocytic monocytes in contrast to a higher number of phagocytic cord blood monocytes only in premature infants. Mycoses, 1998 May-Jun, 41(5-6), 219 - 21 Extensive skin candidosis in an adult: effective treatment with itraconazole; Daning L et al.; A 34-year-old man with 6 years' untreated erythematous scaling of the skin was diagnosed as having extensive skin candidosis . Oral itraconazole was administered for 4 weeks (400 mg day-1 for the first 3 weeks and 200 mg day-1 for the last week) . At the end of the 4-week treatment period, the rash was completely cleared from all skin sites, and Candida albicans was absent in culture tests on tissue samples . No adverse events were reported by the patient, and laboratory analysis revealed no abnormalities in the liver, kidney and haematological systems . Oral itraconazole is therefore an effective treatment for extensive skin candidosis. Gene, 1998 Jul 30, 215(2), 311 - 8 Cloning and characterization of mitochondrial methionyl-tRNA synthetase from a pathogenic fungi Candida albicans; Lee SW et al.; A genomic sequence encoding mitochondrial methionyl-tRNA synthetase (MetRS) was determined from a pathogenic fungi Candida albicans . The gene is distinct from that encoding the cytoplasmic MetRS . The encoded protein consists of 577 amino acids (aa) and contains the class I defining sequences in the N-terminal domain and the conserved anticodon-binding amino acid, Trp, in the C-terminal domain . This protein showed the highest similarity with the mitochondrial MetRSs of Saccharomyces cerevisiae and Shizosaccharomyces pombe . The mitochondrial MetRSs of these fungi were distinguished from their cytoplasmic forms . The protein lacks the zinc binding motif in the N-terminal domain and the C-terminal dimerization appendix that are present in MetRSs of several other species . Escherichia coli tRNAMet was a substrate for the encoded protein as determined by genetic complementation and in vitro aminoacylation reaction . This cross-species aminoacylation activity suggests the conservation of interaction mode between tRNAMet and MetRS. Infect Immun, 1998 Sep, 66(9), 4440 - 9 CD86 (B7-2), but not CD80 (B7-1), expression in the epidermis of transgenic mice enhances the immunogenicity of primary cutaneous Candida albicans infections; Gaspari AA et al.; Transgenic (Tg) mice whose epidermal keratinocytes constitutively overexpress either B7-1 (CD80) or B7-2 (CD86) exhibited exaggerated cutaneous delayed type hypersensitivity (DTH) to haptens compared to non-Tg mice . To determine whether enhanced DTH in these Tg mice is seen in response to cutaneous fungal infections, a primary infection with Candida albicans was established by inoculating this organism on the occluded skin of Tg and non-Tg mice . These infections resolved 7 days after removal of occlusive dressing in all three groups of mice, without evidence of exaggerated inflammation in either the Tg or non-Tg mice . Only B7-2 Tg mice developed enhanced Th1-lymphocyte-mediated immune responses to C . albicans antigens after resolving this infection: enhanced footpad swelling in response to intradermal C . albicans antigens, enhanced production of mRNA encoding Th1 lymphokines in draining lymph nodes, and increased gamma interferon secreted into culture supernatants by lymph node T lymphocytes stimulated with Candida antigens in vitro . Lastly, Western blotting of sera from mice that had resolved this fungal infection indicated that only B7-2 Tg mice recognized a wide range of Candida-associated antigens . These data suggest that these two costimulatory molecules, when expressed by keratinocytes, do not deliver identical signals to C . albicans antigen-reactive Th1 lymphocytes . The enhanced immune response in B7-2 Tg mice to a cutaneous C . albicans infection demonstrates the importance of antigen presentation and costimulation in immune reactivity to fungi . Furthermore, B7-2 Tg mice may be useful in identification of protective Candida antigens. Infect Immun, 1998 Sep, 66(9), 4331 - 9 Coiling phagocytosis of trypanosomatids and fungal cells; Rittig MG et al.; Coiling phagocytosis has previously been studied only with the bacteria Legionella pneumophila and Borrelia burgdorferi, and the results were inconsistent . To learn more about this unconventional phagocytic mechanism, the uptake of various eukaryotic microorganisms by human monocytes, murine macrophages, and murine dendritic cells was investigated in vitro by video and electron microscopy . Unconventional phagocytosis of Leishmania spp . promastigotes, Trypanosoma cruzi trypomastigotes, Candida albicans hyphae, and zymosan particles from Saccharomyces cerevisiae differed in (i) morphology (rotating unilateral pseudopods with the trypanosomatids, overlapping bilateral pseudopods with the fungi), (ii) frequency (high with Leishmania; occasional with the fungi; rare with T . cruzi), (iii) duration (rapid with zymosan; moderate with the trypanosomatids; slow with C . albicans), (iv) localization along the promastigotes (flagellum of Leishmania major and L . aethiopica; flagellum or posterior pole of L . donovani), and (v) dependence on complement (strong with L . major and L . donovani; moderate with the fungi; none with L . aethiopica) . All of these various types of unconventional phagocytosis gave rise to similar pseudopod stacks which eventually transformed to a regular phagosome . Further video microscopic studies with L . major provided evidence for a cytosolic localization, synchronized replication, and exocytic release of the parasites, extending traditional concepts about leishmanial infection of host cells . It is concluded that coiling phagocytosis comprises phenotypically similar consequences of various disturbances in conventional phagocytosis rather than representing a single separate mechanism. FEMS Microbiol Lett, 1998 Aug 1, 165(1), 103 - 9 Identification of four chitin synthase genes in the rice blast disease agent Magnaporthe grisea; Vidal-Cros A et al.; Four chitin synthase gene fragments were isolated from Magnaporthe grisea by use of polymerase chain reaction . A pair of degenerate primers based on conserved zymogen-type chitin synthase sequences amplified a approximately 600-bp product containing three chitin synthase gene fragments . A second pair of degenerate primers, based on conserved sequences between the chitin synthases 3 of Saccharomyces cerevisiae and Candida albicans, amplified a single 770-bp fragment . The four corresponding amino acid sequences each fell into one of the chitin synthase classes I-IV, as deduced from sequence analysis . Northern analysis demonstrated that the class II gene was expressed transiently in early phases of growth, whereas the class I and III genes as well as the class IV gene were expressed throughout the entire life cycle . Furthermore, the class III gene was the most expressed. J Clin Pathol, 1998 May, 51(5), 390 - 1 A novel technique for assessment of adherence of Candida albicans to solid surfaces; Williams DW et al.; A novel approach for the assessment of adherence of Candida albicans to translucent acrylic material is described . The method uses the inverted microscope to visualise yeast adhering to acrylic surfaces while the test material remains immersed in buffer . Adherent cells were not subjected to surface tension forces that can occur during drying processes, so that an even distribution of yeast with no aggregation occurred . The process of counting attached yeast was subsequently performed without difficulty . From the 11 C albicans isolates examined, two groups were evident with respect to acrylic adherence: one group of four isolates with an adherence level of 400 yeast/mm2 acrylic, and one group of seven isolates with adherence levels of 1000 yeast/mm2 acrylic. Aust Dent J, 1998 Jun, 43(3), 160 - 6 Candida-associated denture stomatitis . Aetiology and management: a review . Part 2 . Oral diseases caused by Candida species; Webb BC et al.; Certain systemic conditions and/or defects in the immune system may predispose the host to oral candidal infection and the commonest form of oral candidosis is candida-associated denture stomatitis . Until recently there has been controversy concerning the aetiology of the disease . Although some earlier investigators linked denture stomatitis with trauma or bacterial infection, others had isolated Candida albicans from the mouths of patients with the condition . Current studies indicate that denture stomatitis lesions are associated with the detection of candida species while other factors such as denture hygiene, trauma, systemic diseases and deficiencies of the immune system may be involved. Proc Natl Acad Sci U S A, 1998 Aug 18, 95(17), 9825 - 30 The Candida albicans KRE9 gene is required for cell wall beta-1, 6-glucan synthesis and is essential for growth on glucose; Lussier M et al.; We have isolated CaKRE9, a gene from Candida albicans, that is a functional homologue of the Saccharomyces cerevisiae KRE9 gene involved in beta-1,6-glucan synthesis . Disruption of the CaKRE9 gene in C . albicans shows that CaKre9p is required for the synthesis or assembly of this fungal polymer . Homozygous null disruptants of CaKRE9 grow poorly on galactose and fail to form hyphae in serum, and, in growth medium containing glucose, the gene is essential . Thus, the CaKRE9 gene product is a potentially useful candidate as a target for fungal-specific drugs. Am J Health Syst Pharm, 1998 Aug 1, 55(15), 1584 - 7 Microbial inhibitory properties and stability of topotecan hydrochloride injection; Patel K et al.; The viability of five microorganisms in topotecan 1 mg/mL (as the hydrochloride salt) in sterile water and the stability of the drug were studied . Duplicate portions of topotecan 1 mg/mL were inoculated with Escherichia coli . The process was repeated for Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Aspergillus niger . Samples were removed from each solution initially and after 6, 16, and 24 hours and 3, 7, 14, 21, and 28 days of incubation at 20-25 degrees C . To test stability, vials of reconstituted topotecan hydrochloride injection were stored at each of three temperatures--5, 25, and 30 degrees C--and other vials were used for time zero analysis . For each temperature, vials were removed at 1, 7, and 14 days and the remaining vials at 28 days for analysis by high-performance liquid chromatography and for visual and pH assessment . P . aeruginosa, S . aureus, and E . coli lost viability at 16 hours, 24 hours, and 28 days, respectively . C . albicans and A . niger did not lose viability, but their numbers did not grow . No differences in color or clarity were observed, and pH was constant . In all solutions, the topotecan concentration was > 98% of the initial concentration . Topotecan 1 mg/mL in sterile water stored at 20-25 degrees C for up t 28 days did not support growth of the five microorganisms studied; in solutions stored at 5, 25, or 30 degrees C for up to 28 days, topotecan 1 mg/mL remained stable. Eur Surg Res, 1998, 30(4), 290 - 6 Measurement of (1-->3)-beta-D-glucan in an experimental model of systemic candidiasis; Kawagoe T et al.; To investigate the utility of measuring blood concentrations of (1-->3)-beta-D-glucan, a component of the fungal cell wall, as an auxiliary diagnostic method for systemic candidiasis, rats were inoculated with Candida albicans and the number of C . albicans in the viscera and glucan in the blood were quantitated . The concentration of blood glucan and the number of C . albicans in the viscera were also measured both under leukopenia and with deteriorated reticuloendothelial system cell function, and when the liver and spleen had been excised . As a result, systemic candidiasis appeared in the group with leukopenia, and the number of living C . albicans increased in the kidney and liver . Together with this increase in the number of C . albicans, there was an increase in blood (1--> 3)-beta-D-glucan . Measurements of blood (1--> 3)-beta-D-glucan well reflect a proliferation of C . albicans in vivo, which would make this a useful auxiliary for the clinical diagnosis of systemic mycosis. J Clin Microbiol, 1998 Sep, 36(9), 2690 - 5 Stable phenotypic resistance of Candida species to amphotericin B conferred by preexposure to subinhibitory levels of azoles; Vazquez JA et al.; The fungicidal activity of amphotericin B (AmB) was quantitated for several Candida species . Candida albicans and C . tropicalis were consistently susceptible to AmB, with less than 1% survivors after 6 h of exposure to AmB . C . parapsilosis and variants of C . lusitaniae and C . guilliermondii were the most resistant, demonstrating 50 to 90% survivors in this time period and as high as 1% survival after a 24-h exposure time . All Candida species were killed (<1% survivors) after 24 h of exposure to AmB . In contrast, overnight exposure to either fluconazole or itraconazole resulted in pronounced increases in resistance to subsequent exposures to AmB . Most dramatically, C . albicans was able to grow in AmB cultures after azole preexposure . Several other Candida species did not grow in AmB but showed little or no reduction in viability after up to 24 h in AmB . Depending on the growth conditions, Candida cells preexposed to azoles may retain AmB resistance for days after the azoles have been removed . If this in vitro antagonism applies to the clinical setting, treatment of patients with certain antifungal combinations may not be beneficial . The ability of some Candida isolates to survive transient exposures to AmB was not reflected in the in vitro susceptibility changes as measured by standard MIC assays . This finding should be considered in studies attempting to correlate patient outcome with in vitro susceptibilities of clinical fungal isolates . Patients who fail to respond to AmB may be infected with isolates that are classified as susceptible by standard in vitro assays but that may be resistant to transient antifungal exposures which may be more relevant in the clinical setting. J Ethnopharmacol, 1998 Jul, 61(3), 237 - 41 Antimicrobial and anticonvulsant activities of Viscum capense; Amabeoku GJ et al.; Dichloromethane, methanol and water extracts of Viscum sapense L.f., of the Loranthaceae family, were tested for antimicrobial activities against Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans . Methanol extract was also tested for activity against seizures in albino mice induced by pentylenetetrazole (PTZ), bicuculline and N-methyl-DL-aspartic acid (NMDLA) . Methanol extract of V . capense inhibited the growth of S . aureus . Methanol extract also protected the mice against PTZ- and bicuculline-induced tonic seizures but did not significantly alter NMDLA-induced tonic seizures . The data indicate that the extract of V . capense has antibacterial activity against S . aureus and also anticonvulsant activity. Gynecol Obstet Invest, 1998 Aug, 46(2), 73 - 4 Breast-feeding, pain and infection; Thomassen P et al.; A syndrome of deep pain in the breast during and immediately after lactation has been ascribed to an infection with Candida albicans . A series of 20 patients with deep pain, another 20 with superficial infection and 20 healthy women were compared with respect to the growth of bacteria and fungi . C . albicans was found twice as often in the milk of women with superficial lesions compared to those with deep pain . Bacteria were often found on the nipple and in the milk of those complaining of deep pain . Thus, if the deep pain syndrome is caused by microbes, this study points to a pathogenic role of bacteria rather than fungi. Zh Mikrobiol Epidemiol Immunobiol, 1998 May-Jun, (3), 51 - 5 {Proliferative response and suppressor activity of lymphocytes in candidiasis}; Sardyko NV et al.; In 14 practically healthy donors, 22 patients with chronic candidiasis of the skin and mucosa (CCSM) and 14 patients with chronic candidal vulvovaginitis (CCW) immunoregulating factors synthesized by peripheral blood lymphocytes under the conditions of in vitro induction of suppressor cells were studied in comparison with the proliferative response to PHA and Candida albicans cytoplasmic protein . In practically healthy donors the level of lymphocyte response to mitogen coincided with the presence of the inhibiting activity of supernatants . In a more severe form of Candida infection (CCSM) the proliferative reaction of lymphocytes to both PHA and C.albicans antigen was weakened or even inhibited . The level of the proliferative response of lymphocytes was inversely proportional to the immunoregulating activity of cells, nonstimulated and stimulated with ConA and C.albicans cytoplasmic protein . In CCW lymphocytes exhibited good proliferative response to T-mitogen, occurring in combination with weak reaction, or no reaction at all, to C.albicans antigen . A decrease in the level of spontaneous blast transformation was accompanied by an increase in spontaneous suppressor activity. Dev Comp Immunol, 1998 Jul-Aug, 22(4), 387 - 99 A gloverin-like antibacterial protein is synthesized in Helicoverpa armigera following bacterial challenge; Mackintosh JA et al.; A bacteria inducible antibacterial protein, P2, was isolated from the old world bollworm Helicoverpa armigera . Fifth-instar larvae were injected with live Escherichia coli NCTC 8196 . P2 was isolated by HPLC using reversed-phase and size-exclusion columns . In addition, P2 was isolated by an alternative method of sequential cation-exchange and reversed-phase HPLC . The structure of P2 was determined by N-terminal Edman degradation and mass spectrometry . P2 had similar mass (14.1 kDa) structure and activity to gloverin, an inducible glycine-rich antibacterial protein isolated from Hyalophora gloveri {Axen, A.; Carlsson, A.; Engstrom, A.; Bennich, H . Eur . J . Biochem . 247:614-619; 1997} . At the N-terminus P2 had approximately 60% identity with gloverin . P2 is basic, heat stable, and displayed rapid antibacterial action . P2 was active against the Gram-negative bacteria tested and was inactive against the Gram-positive bacteria, Candida albicans, a bovine turbinate cell line, and pestivirus. J Infect Dis, 1998 Aug, 178(2), 488 - 96 Recognition of antigenic clusters of Candida albicans by T lymphocytes from human immunodeficiency virus-infected persons; Kunkl A et al.; The fine specificity of the cellular immune response to Candida albicans (i.e., recognition of different antigenic components) between normal controls and human immunodeficiency virus-infected patients in various stages of disease was compared . C . albicans-specific T cells, enriched by antigen stimulation and interleukin-2 expansion, were challenged with antigenic fractions of different molecular weight obtained by SDS-gel fractionation of C . albicans extracts in the presence of autologous mononuclear cells as antigen-presenting cells . Proliferative responses showed similar patterns of reactivity between controls and category A and B seropositive subjects . Category C patients with concurrent C . albicans infections did not give rise to C . albicans-specific T cell lines, confirming the T cell defect . Patients without clinically evident C . albicans infection had a low but broad reactivity pattern of C . albicans-specific T cells . These results suggest that depletion of C . albicans-specific T cells, independent of their fine specificity, occurs along with disease progression. J Infect Dis, 1998 Aug, 178(2), 478 - 87 Candidiasis in interferon-gamma knockout (IFN-gamma-/-) mice; Balish E et al.; Germ-free C57BL/6 x 129 interferon-gamma knockout (IFN-gamma(-/-)) mice and their immunocompetent (+/-, +/+) counterparts were colonized with a pure culture of Candida albicans to assess their natural susceptibility to mucosal and systemic candidiasis of endogenous origin . Colonization with a pure culture of C . albicans was not lethal for adult or neonatal IFN-gamma(-/-) gnotobiotic mice over the 15-week study . The IFN-gamma(-/-) mice