J Biol Chem, 1998 Mar 13, 273(11), 6139 - 43 Isolation from an ant Myrmecia gulosa of two inducible O-glycosylated proline-rich antibacterial peptides; Mackintosh JA et al.; Reported here is the isolation and characterization of two antibacterial peptides synthesized in an ant Myrmecia gulosa in response to bacterial challenge . The peptides were purified by reversed-phase high performance liquid chromatography and characterized by peptide sequencing and mass spectrometry . Both peptides were formed from 16 amino acids, were rich in proline ( approximately 30%), and had N-acetylgalactosamine O-linked to a conserved threonine . The activity of a synthetic non-glycosylated isoform was markedly reduced demonstrating that glycosylation was necessary for maximum activity . The peptides were active only against growing Escherichia coli . They were inactive against stationary cells, Gram-positive bacteria, the yeast Candida albicans, two species of mammalian cells, and bovine pestivirus. J Biol Chem, 1998 Mar 6, 273(10), 5638 - 44 Hemoglobin induces binding of several extracellular matrix proteins to Candida albicans . Identification of a common receptor for fibronectin, fibrinogen, and laminin; Yan S et al.; Host infection by the pathogenic fungus Candida albicans is initiated by adhesion and mediated by binding to several host extracellular matrix proteins . Previously, we demonstrated that hemoglobin supplemented into a chemically defined medium significantly and specifically induced fibronectin binding to C . albicans . We now report that hemoglobin also induces binding of laminin, fibrinogen, and type IV collagen but not of thrombospondin-1 or type I collagen . The binding of each protein was inhibited by the respective unlabeled ligand in a concentration-dependent manner . Fibrinogen inhibited the binding of radiolabeled fibronectin, laminin, and fibrinogen with similar IC50 values, suggesting that a single promiscuous receptor recognizes these three proteins . Competitive binding studies indicated that a second class of receptor binds specifically to laminin . Growth of C . albicans in the presence of hemoglobin also increased cell adhesion to immobilized fibronectin, laminin, fibrinogen, and type IV collagen but not to thrombospondin-1 or type I collagen . Exposure to hemoglobin induced increased or de novo expression of several surface proteins on C . albicans . One of these proteins with a molecular weight of 55,000 recognized fibronectin, based on ligand protection and affinity chromatography on immobilized fibronectin . Thus, hemoglobin induces both promiscuous and specific receptors for extracellular matrix proteins and, therefore, may regulate matrix adhesion during dissemination of C . albicans infections. Proc Natl Acad Sci U S A, 1998 Mar 3, 95(5), 2417 - 22 An additional role for the F-box motif: gene regulation within the Neurospora crassa sulfur control network; Kumar A et al.; The F-box represents a protein motif originally identified as a conserved amino-terminal domain within the Neurospora crassa negative regulator sulfur controller-2 . Recently, F-boxes have been found within a number of cell cycle regulatory proteins, where they mediate ubiquitin-driven proteolytic events required for major cell cycle transitions . F-box function, however, is not restricted solely to cell cycle pathways . Here we present evidence expanding F-box function to encompass gene regulatory processes independent of the cell cycle through in vivo analysis of an F-box acting within the N . crassa sulfur regulatory network . The Neurospora sulfur circuit features a set of regulatory genes acting to modulate gene expression based on environmental sulfur conditions . These sulfur regulatory genes include cys-3+, which encodes a basic region-leucine zipper transcriptional activator, as well as the negative regulatory gene scon-2+ . Through site-directed mutagenesis of the SCON2 F-box, we have generated a sulfur auxotrophic phenotype previously unobserved in any scon-2 mutant . Using Northern analysis, we have traced this auxotrophy to a complete shutdown of cys-3+ gene expression . We have further analyzed F-box function by constructing a series of chimeric SCON2 proteins containing swapped F-box domains from the yeast transcriptional inhibitor Met30p and the Candida albicans cell cycle regulator Cdc4p . The ability of these chimeric proteins to restore partial wild-type sulfur regulation in vivo emphasizes the universal nature of this motif and confirms the functional importance of the F-box within noncell cycle regulatory pathways. Zentralbl Bakteriol, 1998 Jan, 287(1-2), 157 - 69 In vitro efficacy of a hydrophilic central venous catheter loaded with silver to prevent microbial colonization; Gatter N et al.; A method was developed to load the surface of a central venous catheter with silver to prevent bacterial colonization . Silver confers a broad antimicrobial activity with a relatively low risk of resistance . Catheters were incubated with a silver nitrate solution in different concentrations . The solvent, incubation temperature and incubation period were varied to examine the influence on the catheter loading . With increasing incubation temperature, time and concentration of silver nitrate, higher rates of silver elution were observed by atomic absorption spectroscopy . Furthermore, by using ethanol-water as a solvent instead of pure water, the amount of silver bound to the catheter surface was enhanced . The release of silver from the catheter surface is mainly controlled by first order kinetics . Antimicrobial efficacy of the modified catheter, in comparison to unloaded catheters, was tested in a stationary and a dynamic model with different microorganisms . Adherence experiments with Candida albicans showed almost complete inhibition of growth during a period of 72 hours, including initial adherence . While initial adherence of bacteria could not be prevented, these experiments showed an excellent reduction of bacterial colonization . In a perfusion model, adhesion of E . coli could be reduced for at least seven days . Further studies are planned to examine prolonged antimicrobial effects. FEMS Microbiol Lett, 1998 Mar 15, 160(2), 191 - 7 Expression of CDR1, a multidrug resistance gene of Candida albicans: transcriptional activation by heat shock, drugs and human steroid hormones; Krishnamurthy S et al.; We have examined the expression of CDR1 (Candida drug resistance gene) in different stress conditions . There was a significant but transient enhancement of CDR1 expression associated with elevated temperatures . Most noteworthy transcriptional activation was observed with miconazole and vinblastine . Interestingly, beta-estradiol and progesterone were also able to enhance CDR1 expression . Elevated levels of CDR1 and CDR2 (a homologue of CDR1) mRNA were found in some azole-resistant clinical isolates of C . albicans . CaMDR1 (benomyl-resistant) expression, however, did not differ among all the resistant isolates . Our results confirm the existence of multiple mechanisms of azole resistance in C . albicans. Can J Neurol Sci, 1998 Feb, 25(1), 76 - 8 Vasculitic basilar artery thrombosis in chronic Candida albicans meningitis; Grimes DA et al.; BACKGROUND: Cerebrovascular complications of meningitis have been most extensively documented in the setting of acute bacterial or chronic tuberculous meningitis . Involvement of major cerebral vessels is rare and basilar artery thrombosis has not been reported in fungal meningitis secondary to candida infection . METHODS: We describe the clinical course and neuropathological findings in a woman with chronic meningitis due to Candida albicans . RESULTS: The diagnosis remained elusive antemortem despite analysis of 7 large volume CSF samples and examination of a meningeal and cortical biopsy . Death followed extensive brainstem and temporo-occipital infarction secondary to basilar artery thrombosis . The basilar artery occlusion was secondary to an intense, granulomatous and necrotizing basal meningitis focally extending to the media and intima . CONCLUSIONS: This paroxysmal and devastating complication of untreated chronic candida meningitis reinforces that a trial of empirical therapy with both antituberculous and antifungal agents should be considered in most cases of chronic culture-negative lymphocytic meningitis. J Chemother, 1998 Feb, 10(1), 7 - 16 The effect of the new triazole, voriconazole (UK-109,496), on the interactions of Candida albicans and Candida krusei with endothelial cells; Fratti RA et al.; In this study, we investigated how voriconazole affects specific endothelial cell interactions utilizing both fluconazole-susceptibles and resistantR Candida albicans strains (C . albicansS and C . albicansR, respectively) as well as Candida krusei . Our data show that exposing C . albicansS to voriconazole significantly reduced its adherence to endothelial cells (p <0.001) . The adherence of C . albicansR to endothelial cells was not affected by treatment with either antifungal agent . Exposure of C . albicans to both agents inhibited germ tube formation; however, voriconazole showed higher ability in inhibiting germination as compared with fluconazole . The effect of antifungals on germination was also tested during co-incubation of yeast cells with endothelial cells . Pretreated C . albicansS cells germinated on endothelial cells in the presence of voriconazole or fluconazole . However, the degree of germination was reduced by 81% and 16%, respectively . Similar results were observed with C . albicansR . Our data demonstrate that voriconazole treatment reduced the median germ tube length of C . albicansS and C . albicansR by approximately 60%, whereas fluconazole reduced the germ tube length of these strains by 27% and 63%, respectively (P < 0.0001 for each comparison) . We compared the efficacy of voriconazole and fluconazole in protecting endothelial cells against damage caused by C . albicansS, C . albicansR, and C . krusei . Voriconazole and fluconazole reduced C . albicans-mediated endothelial cell injury by about 90% and 40%, respectively (P < 0.01 for each comparison) . Additionally, voriconazole treatment significantly reduced C . krusei-mediated injury to endothelial cells by 69% (P < 0.01), whereas fluconazole did not exhibit significant protection (P < 0.6) . These results demonstrate that voriconazole, in addition to its direct inhibitory activity against fungi, may act against Candida spp . by interfering with critical host/parasite interactions, such as adherence and endothelial cell damage, as well as germination . Therefore, this triazole represents a new and promising agent for the treatment of disseminated candidal infections caused by both fluconazole-susceptible and -resistant species. Microbiol Mol Biol Rev, 1998 Mar, 62(1), 130 - 80 Cell wall and secreted proteins of Candida albicans: identification, function, and expression; Chaffin WL et al.; The cell wall is essential to nearly every aspect of the biology and pathogenicity of Candida albicans . Although it was initially considered an almost inert cellular structure that protected the protoplast against osmotic offense, more recent studies have demonstrated that it is a dynamic organelle . The major components of the cell wall are glucan and chitin, which are associated with structural rigidity, and mannoproteins . The protein component, including both mannoprotein and nonmannoproteins, comprises some 40 or more moieties . Wall proteins may differ in their expression, secretion, or topological location within the wall structure . Proteins may be modified by glycosylation (primarily addition of mannose residues), phosphorylation, and ubiquitination . Among the secreted enzymes are those that are postulated to have substrates within the cell wall and those that find substrates in the extracellular environment . Cell wall proteins have been implicated in adhesion to host tissues and ligands . Fibrinogen, complement fragments, and several extracellular matrix components are among the host proteins bound by cell wall proteins . Proteins related to the hsp70 and hsp90 families of conserved stress proteins and some glycolytic enzyme proteins are also found in the cell wall, apparently as bona fide components . In addition, the expression of some proteins is associated with the morphological growth form of the fungus and may play a role in morphogenesis . Finally, surface mannoproteins are strong immunogens that trigger and modulate the host immune response during candidiasis. Mol Gen Genet, 1998 Feb, 257(4), 412 - 20 Expression of a chromosomally integrated, single-copy GFP gene in Candida albicans, and its use as a reporter of gene regulation; Morschhauser J et al.; Genetically engineered versions of the GFP gene, which encodes the green fluorescent protein of Aequorea victoria, were placed under the control of the constitutively active Candida albicans ACT1 promoter and integrated in single copy into the genome of this pathogenic yeast . Integrative transformants in which one of the two ACT1 alleles had been replaced by a GFP gene exhibited a homogeneous, constitutive fluorescent phenotype . Cells expressing GFP with the wild-type chromophore exhibited very weak fluorescence compared to those GFP proteins with the S65T or S65A, V68L, S72A (GFPmut2) chromophore mutations . Substitution of the CTG codon, which specifies serine instead of leucine in C . albicans, by TTG was absolutely necessary for GFP expression . Although GFP mRNA levels in cells containing a GFP gene with the CTG codon were comparable to those of transformants containing GFP with the TTG substitution, only the latter produced GFP protein, as detected by Western blotting, suggesting that the frequent failure to express heterologous genes in C . albicans is principally due to the noncanonical codon usage . Transformants expressing the modified GFP gene from the promoter of the SAP2 gene, which encodes one of the secreted acid proteinases of C . albicans, showed fluorescence only under conditions which promote proteinase expression, thereby demonstrating the utility of stable, chromosomally integrated GFP reporter genes for the study of gene activation in C . albicans. Infect Immun, 1998 Apr, 66(4), 1783 - 6 Expression of the Candida albicans gene ALS1 in Saccharomyces cerevisiae induces adherence to endothelial and epithelial cells; Fu Y et al.; To identify genes encoding adhesins that mediate the binding of Candida albicans to endothelial cells, a genomic library from this organism was constructed and used to transform Saccharomyces cerevisiae . These transformed organisms were screened for adherence to endothelial cells, and a highly adherent clone was identified . The adherence of this clone to endothelial cells was over 100-fold greater than that of control S . cerevisiae transformed with the empty plasmid . This clone also exhibited enhanced adherence to epithelial cells . The C . albicans gene contained within this clone was found to be ALS1 . These results indicate that ALS1 may encode a candidal adhesin. Infect Immun, 1998 Apr, 66(4), 1708 - 17 Characteristics of invasive candidiasis in gamma interferon- and interleukin-4-deficient mice: role of macrophages in host defense against Candida albicans; Kaposzta R et al.; Murine models of invasive candidiasis were used to study the in vivo importance of gamma interferon (IFN-gamma) and interleukin-4 (IL-4) in host defense against Candida albicans and to characterize the tissue inflammatory reactions, with special reference to macrophages (Mphi) . Knockout (KO) IFN-gamma-deficient (GKO) and IL-4-deficient (IL-4 KO) and C57BL/6 parental mouse strains were challenged intraperitoneally with 10(8) C . albicans blastoconidia . Survival of GKO mice was significantly lower (16.7%) than that of C57BL/6 control (55.5%) and IL-4 KO (61.1%) animals, but was not correlated with the extent of organ colonization . Immunohistological analysis with a panel of myeloid and lymphoid markers revealed multiple renal abscesses, myocarditis, hepatitis, meningoencephalitis, and pneumonia in each strain, with a dominant presence of Mphi . In the absence of IFN-gamma, C . albicans induced striking changes in the phenotype of alveolar Mphi and extensive perivascular lymphoid infiltrates in the lung . Impairment in nitric oxide production by peritoneal Mphi was shown only in GKO mice, and they produced Candida-specific immunoglobulin G (IgG), IgM, IgA, and IgG subclasses in lower titers . Our in vivo studies with KO mice elucidate a critical role for IFN-gamma, but not IL-4, in host defense against C . albicans. Infect Immun, 1998 Apr, 66(4), 1392 - 9 Alterations in frequency of interleukin-2 (IL-2)-, gamma interferon-, or IL-4-secreting splenocytes induced by Candida albicans mannan and/or monophosphoryl lipid A; Li SP et al.; We have shown previously that intravenous injection of Candida albicans mannan (MAN) into naive mice induced CD8+ effector downregulatory cells and that such cells were not produced if mice were deficient in CD4+ or I-A+ cells during the early interval (< or =30 h) following the introduction of MAN . Moreover, the nonspecific biological response modifier monophosphoryl lipid A (MPL), given in vivo or incubated with cells in vitro, can abrogate the MAN-specific immunomodulatory activity . The mechanism by which the abrogation is mediated is unknown, but it is hypothesized to involve cytokines . Therefore, we measured the number of cytokine-secreting cells for the Thl cytokine interleukin-2 (IL-2) and the Th2 cytokine IL-4, as well as for gamma interferon (IFN-gamma), in splenocyte populations from MAN and/or MPL-treated mice, using an enzyme-linked immunospot assay designed to detect individual cytokine-secreting cells (spot-forming cells {SFC}) . Cytokine-secreting cells were demonstrated in cell suspensions enriched for CD4+ cells, but no SFC could be demonstrated in populations enriched for CD8+ cells . Both MAN and MPL, when administered to separate groups of animals, stimulated the production of increased numbers of cytokine-producing cells for each of the three cytokines tested . The response with respect to IL-4-secreting cells, however, was the most striking . Despite the fact that MAN and MPL independently caused increases in SFC to all three cytokines, when both MAN and MPL were administered to the same animal, all increases were reversed, and the numbers of SFC detected were at or below those detected in saline control animals . These data support the hypothesis that IL-4 is involved in MAN-specific immunoregulatory activities . The data also emphasize the fact that two immunomodulators, i.e., MAN and MPL, having similar effects when given in vivo independently, may be antagonistic when administered sequentially to the same animal. Infect Immun, 1998 Apr, 66(4), 1384 - 91 Cytokine involvement in immunomodulatory activity affected by Candida albicans mannan; Wang Y et al.; Candida albicans mannoprotein (MAN) administered intravenously to mice stimulates the production of splenic CD8+ effector cells which downregulate delayed hypersensitivity (DH) in immunized mice . Cytokine involvement in the induction and/or elicitation of downregulation was studied by (i) examining murine splenocytes qualitatively for mRNA for interleukin-2 (IL-2), IL-4, IL-10, IL-12p40, and gamma interferon (IFN-gamma), (ii) quantitating splenocyte mRNA for IL-12p40 by quantitative-competitive reverse transcriptase-mediated PCR, and (iii) measuring serum levels of IL-12p40 and IL-12p70 by capture enzyme-linked immunosorbent assay, each performed at selected intervals over 96 h after giving MAN . Further, the effect of in vivo administration of anti-IL-4 on the induction and elicitation of MAN-specific DH in MAN-treated mice was measured . Expression of IL-12p40 mRNA in the spleen was reduced to near 0 during the first 24 h but rebounded thereafter . Transcripts for IL-10 were present throughout the 96-h period, whereas those for IL-4 and IFN-gamma were either weak or undetectable prior to 24 to 48 h . In vivo administration of anti-IL-4 partially abrogated the downregulatory effect of MAN only when given at the time of MAN administration . Serum levels of IL-12p40, but not IL-12p70, were increased by 24 h and maximal at 48 h . The antagonistic effect of IL-12p40 could contribute to the mechanism(s) for downregulation of DH . Moreover, IL-10, IL-4, and/or IFN-gamma, interacting with MAN-activated cells in the absence of biologically active IL-12, may induce the production of CD8+ downregulatory effector cells . Partial abrogation of downregulatory activity in animals treated with anti-IL-4 at the time of induction of such activity lends support to this hypothesis. Br J Cancer, 1998 Mar, 77(6), 1015 - 20 Cross-reactivity between Candida albicans and human ovarian carcinoma as revealed by monoclonal antibodies PA10F and C6; Schneider J et al.; Summary Antibodies against Candida albicans antigenic determinants have been reported to cross-react with human tumour cells . We have found that two monoclonal antibodies, C6 and PA1OF, developed at our laboratory against C . albicans antigenic determinants, cross-react with human ovarian cancer on Western blots and immunohistochemistry . We have subsequently used one of them, PA10OF, to test by means of immunohistochemistry a series of 37 human ovarian carcinomas . Out of 37 tumours, 25 (67.6%) expressed the antigen recognized by PA1OF . The reactivity, however, was concentrated on the subgroup of particularly aggressive, invasive carcinomas in advanced stages of the disease (19 out of 24 positive), whereas the antigen was expressed significantly less (P=0.0007) in the subgroup of much less aggressive stage I tumours of low malignant potential, also called borderline carcinomas (2 out of 13 positive) . This cross-reactivity between C . albicans and ovarian carcinoma seems to be attributable to a common antigenic determinant related to tumour aggressiveness. Antimicrob Agents Chemother, 1998 Feb, 42(2), 389 - 93 Cellular accumulation, localization, and activity of a synthetic cyclopeptamine in fungi; Capobianco JO et al.; A novel synthetic cyclopeptamine, A172013, rapidly accumulated by passive diffusion into Candida albicans CCH442 . Drug influx could not be totally facilitated by the membrane-bound target, beta-(1,3)-glucan synthase, since accumulation was unsaturable at drug concentrations up to 10 microg/ml (about 1.6 x 10(-7) molecules/cell), or 25x MIC . About 55 and 23% of the cell-incorporated drug was associated with the cell wall and protoplasts, respectively . Isolated microsomes contained 95% of the protoplast-associated drug, which was fully active against glucan synthesis in vitro . Drug (0.1 microg/ml) accumulation was rapid and complete after 5 min in several fungi tested, including a lipopeptide/cyclopeptamine-resistant strain of C . albicans (LP3-1) . The compound penetrated to comparable levels in both yeast and hyphal forms of C . albicans, and accumulation in Aspergillus niger was 20% that in C . albicans . These data indicated that drug-cell interactions were driven by the amphiphilic nature of the compound and that the cell wall served as a major drug reservoir. Antimicrob Agents Chemother, 1998 Feb, 42(2), 241 - 53 Amino acid substitutions in the cytochrome P-450 lanosterol 14alpha-demethylase (CYP51A1) from azole-resistant Candida albicans clinical isolates contribute to resistance to azole antifungal agents; Sanglard D et al.; The cytochrome P-450 lanosterol 14alpha-demethylase (CYP51A1) of yeasts is involved in an important step in the biosynthesis of ergosterol . Since CYP51A1 is the target of azole antifungal agents, this enzyme is potentially prone to alterations leading to resistance to these agents . Among them, a decrease in the affinity of CYP51A1 for these agents is possible . We showed in a group of Candida albicans isolates from AIDS patients that multidrug efflux transporters were playing an important role in the resistance of C . albicans to azole antifungal agents, but without excluding the involvement of other factors (D . Sanglard, K . Kuchler, F . Ischer, J.-L . Pagani, M . Monod, and J . Bille, Antimicrob . Agents Chemother . 39:2378-2386, 1995) . We therefore analyzed in closer detail changes in the affinity of CYP51A1 for azole antifungal agents . A strategy consisting of functional expression in Saccharomyces cerevisiae of the C . albicans CYP51A1 genes of sequential clinical isolates from patients was designed . This selection, which was coupled with a test of susceptibility to the azole derivatives fluconazole, ketoconazole, and itraconazole, enabled the detection of mutations in different cloned CYP51A1 genes, whose products are potentially affected in their affinity for azole derivatives . This selection enabled the detection of five different mutations in the cloned CYP51A1 genes which correlated with the occurrence of azole resistance in clinical C . albicans isolates . These mutations were as follows: replacement of the glycine at position 129 with alanine (G129A), Y132H, S405F, G464S, and R467K . While the S405F mutation was found as a single amino acid substitution in a CYP51A1 gene from an azole-resistant yeast, other mutations were found simultaneously in individual CYP51A1 genes, i.e., R467K with G464S, S405F with Y132H, G129A with G464S, and R467K with G464S and Y132H . Site-directed mutagenesis of a wild-type CYP51A1 gene was performed to estimate the effect of each of these mutations on resistance to azole derivatives . Each single mutation, with the exception of G129A, had a measurable effect on the affinity of the target enzyme for specific azole derivatives . We speculate that these specific mutations could combine with the effect of multidrug efflux transporters in the clinical isolates and contribute to different patterns and stepwise increases in resistance to azole derivatives. Enferm Infecc Microbiol Clin, 1997 Nov, 15(9), 482 - 4 {Ischemia of the lower limbs as the initial manifestation of Candida albicans endocarditis in a parenteral drug addict}; Barreiro PM et al.; BACKGROUND: Multiple infective complications have been described in injection drug users (IDUs) . Infective endocarditis, most frequently caused by Gram positive bacteria, with classical features, is one of the most dangerous . In a few patients fungi are the cause (less than 5%), and these develop an unusual clinical picture . METHODS: An IDUs patient was admitted in our Hospital for subacute arterial ischemia at the inferior limbs . A mass inside the abdominal aorta was detected by echography and arteriography, which was removed surgically a few hours later . RESULTS: The pathologic evaluation of the surgical specimen revealed its fungal composition; the culture of this material was characteristic of Candida albicans . The clinical suspicion of aortic endocarditis, as the emboligenic source responsible of the inferior limbs ischemia, was confirmed with the performance of an echocardiography . A few hours after surgery the patient got worse; 24 hours later he died due to uncontrolled bleeding of the surgical suture in the aorta . CONCLUSIONS: Fungal endocarditis should be thought in IDUs patients presenting inferior limbs ischemia . Due to the high mortality of this disease, as soon as the diagnosis is suspected, urgent medical and surgical therapy should be started. J Med Chem, 1998 Mar 12, 41(6), 996 - 1000 Novel biologically active nonpeptidic inhibitors of myristoylCoA:protein N-myristoyltransferase; Devadas B et al.; A new class of biologically active nonpeptidic inhibitors of Candida albicans NMT has been synthesized starting from the octapeptide ALYASKLS-NH2 (2) . The synthetic strategy entailed the preparation of novel protected Ser-Lys mimics 9 and 12 from (S)- or (R)-3-iodotyrosine and then grafting key enzyme recognition elements in a stepwise manner . Like 2, compounds 16, 17, and 18 are competitive Candida NMT inhibitors that bind to the peptide recognition site of the enzyme . Moreover, 16-18 have an affinity comparable to that of 2 even though they are devoid of peptide bonds . In contrast to 2, these nonpeptidic inhibitors exhibit antifungal activity. Clin Infect Dis, 1998 Mar, 26(3), 642 - 5 Risk factors for candidemia in a children's hospital; MacDonald L et al.; Candida species are increasingly important nosocomial pathogens in critically ill children . A 2.3-fold increase in the rate of nosocomial candidemia at our 200-bed tertiary care children's hospital prompted a study to identify risk factors for this infection . Twenty-six cases were identified between 1992 and 1993, representing 21% of all nosocomial bloodstream infections . Candida albicans was the most frequent isolate (58%), followed by Candida parapsilosis (27%) . A case-control study revealed that there was a statistically significant association between the occurrence of candidemia and placement of a central venous catheter in the femoral vein (P = .03), the use of a tunneled central venous catheter (P = .05), and prolonged hyperalimentation (P = .04) . Patients with candidemia also were noted to have candiduria more often than controls (P = .003) and were more likely to have had topical antifungal agents prescribed (P = .04) . Multivariate analysis showed that hyperalimentation was an independent risk factor for the development of candidemia . We conclude that measures must be taken to reduce these risk factors whenever possible. Eur J Biochem, 1998 Mar 1, 252(2), 245 - 52 Isolation and characterisation of cAMP-dependent protein kinase from Candida albicans . Purification of the regulatory and catalytic subunits; Zelada A et al.; cAMP-dependent protein kinase (PKA) from Candida albicans yeast cells was isolated and characterised . Structural parameters of the holoenzyme and those of its subunits suggested that C . albicans PKA is a tetramer of 287 kDa composed of two regulatory (R) subunits of 64 kDa and two catalytic (C) subunits of unusually large molecular mass of 78 kDa . The apparent Km for ATP and Kemptide were 30 microM and 60 microM respectively . The {A}0.5 for cAMP activation was 150 nM with a Hill coefficient of 1.6 . The holoenzyme undergoes autophosphorylation on the R subunit, a characteristic of the type-II R subunits . Photoaffinity labeling with 8-azido-{32P}cAMP of crude extracts from yeast and mycelial cells strongly suggests that only one type of R subunit is present in the fungus . The R subunit was purified to apparent homogeneity as a protein of 64 kDa . A highly specific polyclonal antiserum raised against the purified protein immunoprecipitated a 64-kDa protein from crude extracts, indicating that the purified R subunit very probably represents the native form of the protein . The 78-kDa form of the C subunit was detected in crude extracts and in Mono Q Sepharose column fractions with heterologous anti-C Ig . It could be isolated by cAMP treatment of the holoenzyme immunoprecipitated from crude extracts with anti-R serum, but this form could not be purified further . Instead, a 60-kDa protein with the main characteristics of C subunit was purified to near homogeneity from soluble extracts of yeast cells . Evidence is presented that this protein very probably derives from the 78-kDa form by proteolytic degradation. P N G Med J, 1995 Sep, 38(3), 163 - 71 Prevalence of vaginal infections with bacterial vaginosis, Trichomonas vaginalis and Candida albicans among pregnant women at the Port Moresby General Hospital Antenatal Clinic; Klufio CA et al.; A clinico-sociodemographic and microbiological survey was carried out at the Port Moresby General Hospital Antenatal Clinic to determine the prevalences of bacterial vaginosis, Trichomonas vaginalis and Candida albicans vaginal infections in pregnancy and to examine if the infections had any association with some suspected sociodemographic risk factors . The study was carried out between December 1990 and January 1991 . Of 206 consecutive subjects surveyed, 79 (38%) had symptomatic infection . However, on speculum examination, abnormal discharge was seen in 188 (91%) . 118 (57%) had microbiologically confirmed infection . The prevalences of the individual infections were T . vaginalis 19%, C . albicans 23% and bacterial vaginosis 23% . Combined infection, i.e . two infections occurring together in the same subject, was uncommon . None of the infections had an association with any of the sociodemographic characteristics studied . Of the 118 positive subjects, 52 (44%) complained of vaginal discharge and 55 (47%) complained of pruritusPIP: The prevalences of vaginal infections with Trichomonas vaginalis, bacterial vaginosis, and Candida albicans were investigated in 206 consecutive pregnant women presenting to Port Moresby (Papua New Guinea) General Hospital in 1990-91 for their first antenatal visit . Bacteriologic investigation identified Candida in 48 women (23%), T . vaginalis in 39 (19%), and bacterial vaginosis in 48 (23%) . Overall, 118 women (57%) were bacteriologically positive for at least one infection . 79 (38%) of the infected women complained of a vaginal discharge and 78 (38%) reported vulvar irritation; however, vaginoscopy revealed abnormal discharge in 188 (91%) of women with an infection . Infection was not associated with gestational age or any of the sociodemographic variables examined (age, parity, ethnic group, residence, husband's education) . The fact that the majority of pregnant women in this series had a vaginal infection is alarming in light of the hypothesized association of such infections with intra-amniotic infection, endometritis, premature rupture of the membranes, preterm labor or birth, and low birth weight . A randomized, controlled prospective study is needed to assess the extent to which, if any, these infections are related to the high perinatal morbidity and mortality from low birth weight at Port Moresby General Hospital . Ugeskr Laeger, 1998 Mar 9, 160(11), 1627 - 32 {Transjugular intrahepatic portosystemic shunt in the treatment of portal hypertension}; Astrup LB et al.; The transjugular intrahepatic portosystemic shunt (TIPS) represents an important advance in the treatment of complications of portal hypertension . The results from the first 10 TIPS procedures in Arhus are reported . We found, as also documented in other clinical series, that TIPS is more effective in controlling acute haemorrhage than treatment with sclerotherapy and specific medical treatment . Seven out of 10 were treated for acute haemorrhage, and two patients were treated for recurrent variceal bleeding in spite of at least 20 procedures of sclerotherapy and pharmaceutical therapy . One patient was treated with TIPS due to refractory ascites . All 10 TIPS procedures were satisfactory, in four patients it was necessary to embolize collaterals . There were no acute complications associated to the TIPS procedures, but one patient developed stenosis of the shunt within one year, and another chronic encephalopathy . Two patients died, one because of sepsis with Candida albicans, and the other of intracerebral bleeding 16 months after the TIPS procedure. Can J Microbiol, 1998 Jan, 44(1), 74 - 9 In vitro antifungal activity of some Mannich bases of conjugated styryl ketones; Manavathu EK et al.; Four Mannich bases of some conjugated styryl ketones IIa-IId were examined for antifungal activity . These compounds were designed as thiol-alkylators and had two centers for attack by cellular thiols . The most potent compounds IIa and IIb possessed hydrophobic, electron-attracting substituents in the aryl rings and in general had minimum inhibitory concentration (MIC) values of 0.2-25 microM against a variety of fungi . None of the four compounds inhibited the growth of a number of bacteria (MIC > 100 microM) . The minimum fungicidal concentration (MFC) values for IIa and IIb were generally either similar or twofold higher than the MIC figures for fungi . Compound IIa demonstrated rapid, concentration-dependent inhibition of the growth of Candida albicans B311 . The toxicity of IIa to normal human cells was much lower than the concentrations of this compound required to inhibit fungal growth . In summary, this study of four prototypic molecules has revealed that this class of compounds may have potential for further development as candidate antifungal agents. Biochemistry, 1998 Mar 10, 37(10), 3351 - 7 A high-affinity inhibitor of yeast carboxypeptidase Y is encoded by TFS1 and shows homology to a family of lipid binding proteins; Bruun AW et al.; A 25-kDa inhibitor of the vacuolar enzyme carboxypeptidase Y from Saccharomyces cerevisiae has been characterized . The inhibitor, Ic, binds tightly with an apparent Ki of 0.1 nM . Consistent with a cytoplasmic localization, Ic is soluble and contains no sequences which could serve as potential signals for transport into the endoplasmic reticulum . Surprisingly, Ic is encoded by TFS1, which has previously been isolated as a high-copy suppressor of cdc25-1 . CDC25 encodes the putative GTP exchange factor for Ras1p/Ras2p in yeast . In an attempt to rationalize this finding, we looked for a physiological relationship by deleting or overexpressing the gene for carboxypeptidase Y in a cdc25-1 strain . However, this did not change the phenotype of this mutant strain . Ic is the first member of a new family of protease inhibitors . The inhibitor is not hydrolyzed on binding to CPY . It has fairly high degree of specificity, showing a 200-fold higher Ki toward a carboxypeptidase from Candida albicans which is highly homologous to carboxypeptidase Y . The TFS1 gene product shows extensive similarity to a class of proteins termed "21-23-kDa lipid binding proteins", members of which are found in several higher eukaryotes, including man . These proteins are highly abundant in some tissues (e.g., brain) and have in general been found to bind lipids . Considering their homology to Ic, it is tempting to speculate that they may also be inhibitors of serine carboxypeptidases. J Immunol Methods, 1997 Dec 29, 210(2), 227 - 34 A rapid evaluation of phagocytosis and killing of Candida albicans by CD13+ leukocytes; Saresella M et al.; Flow cytometry can be adopted for routine monitoring of the immune functions of human polymorphonuclear leukocytes (PMNs) in several disease states . We recently developed a rapid and reproducible assay for the evaluation of the phagocytosis and killing of Candida albicans blastospores by human PMNs . Whole blood leukocytes were incubated with opsonized fluorescein isothiocyanate-labeled (FITC-labeled) blastospores for phagocytosis and killing assays . To discriminate between ingested, membrane-bound and free C . albicans blastospores, ethidium bromide (EtBr) was added to the samples prior to the flow cytometric analysis . EtBr induces a loss of green fluorescence in non-phagocytized C . albicans blastospores . Phagocytosis is determined by gating the phagocytes and calculating the percentage of phagocyte-associated green fluorescent cells . Intracellular killing is determined by first lysing phagocytes by hypotonic shock and then adding propidium iodide (PI) in order to identify red dead blastospores . Killing is measured in terms of the percentage of double-marked blastospore cells . We suggest that this method is a reliable and inexpensive technique to evaluate the immune reactivity of PMNs and peripheral blood monocytes (PBMs) in cases of immunosuppression. Rev Clin Esp, 1997 Dec, 197(12), 799 - 803 {Current treatment of candidemia in non-neutropenic patients . Amphotericin B or fluconazole? A retrospective study of 62 consecutive patients}; Cobo Reinoso P et al.; BACKGROUND: To analyze the epidemiologic characteristics of non-neutropenic patients with candidemia in a general hospital and the advantages and disadvantages of treatment with amphotericin B or fluconazole . PATIENTS AND METHODS: A total of 62 adult non-neutropenic patients with candidemia and treated with amphotericin B (n = 35) or fluconazole (n = 27) were studied . All episodes were considered to be associated with infection in a vein catheter . The demographic characteristics, risk factors for the development of candidemia, Candida species recovered from blood culture, underlying diseases, and clinical manifestations in both groups were compared . The evolution regarding secondary effects developed with both drugs, therapy failures, long term complications, and overall mortality rate associated with candidemia were analyzed . RESULTS: Both groups were comparable with the exception of the percentage of patients infected with species different from Candida albicans, which was higher in the group of patients who received amphotericin B (57%) than in the fluconazole group (26%) (p = 0.02), and in that patients with severe renal failure or AIDS had received preferentially fluconazole . There were no statistically significant differences regarding the evolution of patients treated with amphotericin B or fluconazole with the following factors: therapy failure (27% versus 19%; p = 0.7), overall mortality rate (40% versus 44%; p = 0.6), and mortality directly related to candidemia (33% versus 30%) . Mortality was significantly higher among patients who had not their vein catheters removed early (78%) compared with those who had their vein catheters removed early (34%) (p = 0.01) . Sixty-six percent of patients treated with amphotericin developed some severe secondary effect, whereas no patient in the fluconazole group developed such effects . CONCLUSIONS: Both amphotericin B and fluconazole seem to be effective drugs for the treatment of vein catheter related candidemia in the non-neutropenic patient, although fluconazole is far less toxic . The early removal of the vein catheter plays a prognostic role with at least the same relevance than the type of antifungal therapy chosen. FEMS Immunol Med Microbiol, 1998 Jan, 20(1), 55 - 67 Fucose-specific adhesins on germ tubes of Candida albicans; Vardar-Unlu G et al.; Lectin-like adhesins of hyphal-form Candida albicans were investigated by conventional fluorescence microscopy, fluorescence microscopy with image analysis, spectrofluorimetry and flow cytometry . Labelling was done with neoglycoprotein probes consisting of sugars (fucose, mannose, glucose, galactose, lactose) covalently linked to bovine serum albumin (BSA), which itself was labelled with fluorescein . The fucose probe bound to both the yeast and germ-tube portions of hyphal-form cells, not especially at the tip, but in the adjacent region of the germ-tube portion . Probes with the other sugars did not label the hyphal-form cells . Fucose-probe binding to the cells was optimal at pH 5.0 in citrate buffer, and was a time-dependent reaction requiring 30-60 min and reaching saturation concentration at 100 microg ml(-1) . Each hyphal-form cell of C . albicans grown in 199 medium was calculated to have about 2 x 10(7) fucose probe-binding sites . There appeared to be no requirement for Ca2+ or Mg2+ in binding . Binding of the fucose probe to the hyphal-form cells was higher at 37 degrees C than at 22 degrees C or 4 degrees C . Fluorescence intensity of the fucose-labelled yeast forms was not increased over the hyphal-form cells . A germ-tube-deficient mutant when exposed to hyphal-form growth conditions for 2 h showed much less binding of the fucose probe than the wild-type which produced germ tubes . Confirmation of specificity and the need for a carrier molecule was obtained by showing that Fuc-BSA (without fluorescein) effectively inhibited the binding of the fucose probe, although L-fucose itself was inactive, as was Gal-BSA. Stomatologiia (Mosk), 1998, 77(1), 48 - 9 {The microorganism count on impressions after their disinfection by submersion in sodium hypochlorite solutions}; Koshmanova TN et al.; The virucidal, bactericidal, and fungicidal activity of sodium hypochlorite is studied with silicone imprints . Poliomyelitis virus (type I vaccine strain Sabin LSc 2 ab with titer 10(7.36) TCD50/ml), bacteriophage f52 with titer 2.10(7) PFU/ml, Staphylococcus aureus strain 906, and Candida albicans in concentrations 10(7) corpuscles/ml in the presence of protein die completely in 20 min when submerged in 0.5% sodium hypochlorite solution . Imprints from alginate materials are destroyed if submerged in this solution. J Antimicrob Chemother, 1998 Jan, 41(1), 59 - 65 In-vitro interaction of terbinafine with amphotericin B, fluconazole and itraconazole against clinical isolates of Candida albicans; Barchiesi F et al.; A chequerboard titration broth microdilution method, performed according to the recommendations of the National Committee for Clinical Laboratory Standards, was applied to study the in-vitro interaction of terbinafine with amphotericin B, fluconazole and itraconazole against 30 strains of Candida albicans isolated from the oral cavities of AIDS patients . MICs were determined spectrophotometrically at 490 nm and read at either 24 h or 48 h . The end-point was defined as the drug concentration resulting in > or = 90% inhibition of growth relative to control growth . Synergy, defined as a fractional inhibitory concentration (FIC) index of < or = 0.50, was observed in 93% (28 of 30) of terbinafine-amphotericin B interactions, in 47% (14 of 30) of terbinafine-fluconazole interactions and in 43% (13 of 30) of terbinafine-itraconazole interactions; antagonism (FIC > 2.0) was not observed . Where synergy was not achieved, there was still a decrease, although not as dramatic, in the MIC of one or both drugs when used in combination . Reading the MICs on day 2 did not significantly affect the mode of interaction of terbinafine-triazoles, while for terbinafine-amphotericin B the proportion of synergic interactions dropped from 93% (28 of 30) to 30% (nine of 30; P = 0.0001) . Antagonism was not observed for any drug combination even at 48 h . Minimum fungicidal concentrations (MFCs) of all drugs alone and in combination were determined against five isolates . Neither terbinafine nor the two triazoles showed fungicidal activity when tested alone or in combination . The fungicidal activity of amphotericin B was slightly enhanced when combined with terbinafine, there being a decrease of two-fold dilutions in the amphotericin B MFCs against all five isolates tested . Thus terbinafine enhances the activities of amphotericin B and triazoles against C . albicans in vitro . Clearly, clinical studies are warranted to elucidate further the potential utility of these combination therapies. Farmaco, 1997 Aug-Sep, 52(8-9), 531 - 7 Quinoxaline chemistry . Part 8 . 2-{Anilino}-3-{carboxy}-6(7)-substituted quinoxalines as non classical antifolate agents . Synthesis and evaluation of in vitro anticancer, anti-HIV and antifungal activity; Loriga M et al.; Thirty quinoxalines bearing a substituted anilino group on position 2, a carboethoxy or carboxy group on position 3 and a trifluoromethyl group on position 6 or 7 of the heterocycle were prepared in order to evaluate in vitro anticancer activity . Preliminary screening performed at NCI showed that most derivatives exhibited a moderate to strong growth inhibition activity on various tumor panel cell lines between 10(-5) and 10(-4) molar concentrations . Interesting selectivities were also recorded between 10(-8) and 10(-6) M for a few compounds . One single compound exhibited good activity against Candida albicans. Biochim Biophys Acta, 1998 Jan 15, 1382(1), 5 - 7 Domain organisation in phosphomannose isomerases (types I and II); Jensen SO et al.; Phosphomannose isomerase (PMI) types I and II were found to possess a conserved protein motif . This motif coincides with the catalytic site of the Candida albicans type I PMI, indicating a common catalytic process for both PMI types . The type II PMI are bifunctional enzymes possessing PMI and guanosine diphospho-D-mannose pyrophosphorylase (GMP) activity in separate catalytic domains, which in some species may function as separate proteins. Curr Top Med Mycol, 1997 Dec, 8(1-2), 43 - 55 Germ tube growth of Candida albicans; Gow NA; The clinical pathogen Candida albicans is a budding yeast that is capable of forming a range of polarized and expanded cell shapes from pseudohyphae to true nonconstricted hyphae . Filamentous forms consist of contiguous uninucleated compartments that are partitioned by septa . It has long been held that the so-called "dimorphic transition" from a budding to a filamentous form may aid the fungus to penetrate epithelia and may therefore be a virulence factor . This review summarized new information regarding the physiology and ecology of hyphal growth in C . albicans . New evidence has demonstrated that hyphae of C . albicans have a sense of touch so that they grow along grooves and through pores (thigmotropism) . This may aid infiltration of epithelial surfaces during tissue invasion . Hyphae are also aerotropic and can form helices when contacting solid surfaces . Growing evidence supports the view that hyphal growth is a response to nutrient deprivation, especially low nitrogen and that filamentous growth enables the fungus to forage for nutrients more effectively . Further insights into the growth of C . albicans have come from the analysis of genes and mutations of Saccharomyces which have begun to reveal the molecular mechanisms underlying the mechanisms of bud site selection, cell polarity and signal transduction pathways that lead to pseudohyphal development in this and other organisms . For example, it is now clear that a MAP-kinase cascade, homologous to the mating pathway in Saccharomyces, regulates filamentous growth in both fungi . However, this must be only one of several overlapping or separate signal transduction pathways for hyphal development because filamentous growth still occurs in mutants of Candida and Saccharomyces which are blocked in this pathway . Cell cycle analyses have shown that hyphal phase cell cycle of Candida is distinct from that in budding and pseudohyphal formation and so pseudohyphal growth of Saccharomyces is not a true model of germ tube growth in Candida . Pseudohyphal growth in both Candida and Saccharomyces involves synchronous division of mother cells and their daughters . In contrast, during germ tube growth of Candida, cytoplasm is unequally partitioned at cytokinesis so that apical cells inherit more cytoplasm and sub-apical cells have a single nucleus but are extensively vacuolated . As a result, apical cells grow and divide while sub-apical cells are apparently arrested in the cell cycle until they can regenerate sufficient cytoplasm to re-enter the cell cycle . Although current studies still fall short of verifying the status of yeast-hypha dimorphism as a virulence factor, they suggest that the cell biology of germ tube growth of C . albicans is well suited for the invasive growth of the fungus in vivo. Curr Top Med Mycol, 1997 Dec, 8(1-2), 15 - 25 Candida dubliniensis: an emerging opportunistic pathogen; Sullivan D et al.; The incidence of opportunistic fungal infections continues to increase, partly as a result of the continuing AIDS epidemic . Candida albicans remains the most important fungal pathogen and is frequently associated with oral candidiasis in HIV-infected individuals . Over the past decade, however, there has been an increasing number of reports implicating other Candida species, such as C . tropicalis, C . glabrata and C . krusei, in disease in these patients and in other patient groups . During the same period there have also been frequent reports in the literature describing what have generally been termed "atypical" C . albicans strains . These isolates have usually been recovered from symptomatic HIV-infected individuals and are unidentifiable as any recognized Candida species using conventional criteria . Two such groups of isolates recovered from cases of oral candidiasis in Irish and Australian HIV-infected and AIDS patients have been postulated to constitute a novel species which has been termed C . dubliniensis . These isolates are phenotypically very similar to C . albicans in that they produce germ tubes and chlamydospores . However, they have unusual carbohydrate assimilation patterns and grow poorly or not at all at 42 degrees C . Using a variety of DNA fingerprinting techniques and karyotype analysis, the genomic organization of C . dubliniensis was shown to be distinctly different from that of C . albicans . Classification of C . dubliniensis as a separate species was confirmed by phylogenetic analysis, whereby the comparison of ribosomal RNA sequences demonstrated that C . dubliniensis isolates formed a cluster clearly distinct from other Candida species, including C . albicans, to which it is most closely related . Since its original identification, atypical Candida isolates from around the world have been positively identified as belonging to this species . To date, isolates of C . dubliniensis have been recovered mainly from the oral cavities of HIV-infected individuals and are most frequently implicated in cases of recurrent infection following antifungal drug treatment . The clinical importance of this species and the role of drug resistance in its epidemiology have yet to be determined. Curr Top Med Mycol, 1996 Dec, 7(1), 71 - 86 Genomic variation in C . albicans; Wickes BL et al.; Candida albicans displays many types of variation which affect a broad spectrum of phenotypes . Among them are antigenic, chromosomal, morphologic, and biochemical variation . The ability to modulate many phenotypes is clearly an important factor in the success of this fungus as a pathogen and variation at the genomic level may be the common denominator among the different systems . Genomic variation in C . albicans has been studied by many researchers and a number of different mechanisms have been identified . Among them are ploidy fluctuations, which allow the organism to cycle from 2n chromosome number to 4n or higher; translocation, which has been demonstrated to involve many different chromosomes and affects many phenotypes including virulence; mitotic recombination, which has been demonstrated to increase resistance to certain drugs; and nondisjunction, which has been shown to have morphological consequences . The number and diversity of these mechanisms combine to make C . albicans a highly successful organism . Although normally a commensal of humans, when invasive, C . albicans can inhabit almost any site in the body . It is not known what governs the transition of C . albicans from a commensal to pathogenic invader, however, variation at the genomic level likely plays a role . One possible consequence of variation is the generation of atypical strains, further expanding the documented phenotypic plasticity of this organism . The exposure of patients to cytotoxic drugs during treatment of such diseases as AIDS or cancer increases the selective pressure and has exacerbated both the frequency and degree of variability observed in C . albicans . The molecular analysis of genomic variation in C . albicans is proving to be a fertile area of research and future investigations can only be expected to add to the mechanisms documented in this review. Curr Top Med Mycol, 1996 Dec, 7(1), 55 - 69 Candida albicans secreted aspartyl proteinases; Hube B; Evidence suggests that infections with the opportunistic yeast Candida albicans are caused by several factors . Among these virulence attributes, secreted aspartyl proteinases (Saps) are widely believed to play a role during pathogenesis . Sap isoenzymes are encoded by at least eight closely related SAP genes . Antigen-antibody studies provided evidence that Sap isoenzymes are expressed in vivo and experimental infections with proteinase deficient mutants suggested a role for Saps in the virulence of C . albicans . However, only one gene product, Sap2, has been characterized in detail . In vitro studies with purified Sap(2) suggested several possible host targets but the role of each Sap isoenzyme remains unclear . The expression pattern of SAP genes proposed that Sap isoenzymes are secreted simultaneously with morphological changes such as the yeast to hyphal transition or during phenotypic switching . In addition, extracellular proteolytic activity may affect adhesion to host cells and thus may help the fungus to persist on host surfaces and to penetrate into deeper tissue . This review will deal with secretory proteinases from C . albicans as putative virulence factors and will focus on the more recent molecular aspects of the proteinases and their genes . Insights into the genetic organization and regulation of the secreted proteinases suggest not only that these enzymes may act as virulence factors of C . albicans, but that the pathogenesis of this fungus is indeed complex and multifactorial. Biochem Mol Biol Int, 1998 Jan, 44(1), 19 - 27 RAPD analysis of Candida albicans strains recovered from different immunocompromised patients (ICP) reveals an apparently non-random infectivity of the strains; Gyanchandani A et al.; The opportunistic imperfect fungus Candida albicans causing life-threatening infections in immunocompromised patients (ICP), especially in HIV-positive cases, is recognized to be one of the most important nosocomial pathogens in the recent decades . The extent of strain-to-strain variation within a species and its relationship to the ability of the organism to colonize or invade a specific group of patients or even a body site is, however, not well known . We have analysed 19 strains of C . albicans recovered from ICP at different locales and times, employing the RAPD technique . No two strains generated identical RAPD profiles with any of the 21 primers tested . Further, the UPGMA clustering of the strains seemingly reflected a certain relationship or nonrandomness in the infection of the patients with the strain of C . albicans vis-a-vis the immunocompromised status due to underlying disease such as diabetes, cancer, asthma and meningitis . These results may have a profound impact on the management of candidiasis, especially in the ICP. Bioorg Med Chem, 1998 Jan, 6(1), 103 - 8 Azole derivatives of 1,4-benzothiazine as antifungal agents; Fringuelli R et al.; A series of azole derivatives of 1,4-benzothiazine 7-14 was synthesized and evaluated for the in vitro and in vivo activity against Candida albicans . Secondary alcohol 10 and its ether derivative 13 showed very good efficacy against systemic candidiasis in a murine experimental model. Nat Struct Biol, 1998 Mar, 5(3), 213 - 21 Crystal structure of the anti-fungal target N-myristoyl transferase; Weston SA et al.; N-myristoyl transferase (NMT) catalyzes the transfer of the fatty acid myristate from myristoyl-CoA to the N-terminal glycine of substrate proteins, and is found only in eukaryotic cells . The enzyme in this study is the 451 amino acid protein produced by Candida albicans, a yeast responsible for the majority of systemic infections in immuno-compromised humans . NMT activity is essential for vegetative growth, and the structure was determined in order to assist in the discovery of a selective inhibitor of NMT which could be developed as an anti-fungal drug . NMT has no sequence homology with other protein sequences and has a novel alpha/beta fold which shows internal two-fold symmetry, which may be a result of gene duplication . On one face of the protein there is a long, curved, relatively uncharged groove, at the center of which is a deep pocket . The pocket floor is negatively charged due to the vicinity of the C-terminal carboxylate and a nearby conserved glutamic acid residue, which separates the pocket from a cavity . These observations, considered alongside the positions of residues whose mutation affects substrate binding and activity, suggest that the groove and pocket are the sites of substrate binding and the floor of the pocket is the catalytic center. Science, 1998 Feb 27, 279(5355), 1355 - 8 Linkage of adhesion, filamentous growth, and virulence in Candida albicans to a single gene, INT1; Gale CA et al.; Adhesion and the ability to form filaments are thought to contribute to the pathogenicity of Candida albicans, the leading cause of fungal disease in immunocompromised patients . Int1p is a C . albicans surface protein with limited similarity to vertebrate integrins . INT1 expression in Saccharomyces cerevisiae was sufficient to direct the adhesion of this normally nonadherent yeast to human epithelial cells . Furthermore, disruption of INT1 in C . albicans suppressed hyphal growth, adhesion to epithelial cells, and virulence in mice . Thus, INT1 links adhesion, filamentous growth, and pathogenicity in C . albicans and Int1p may be an attractive target for the development of antifungal therapies. J Biol Chem, 1998 Feb 20, 273(8), 4492 - 6 The N-terminal membrane domain of yeast NADPH-cytochrome P450 (CYP) oxidoreductase is not required for catalytic activity in sterol biosynthesis or in reconstitution of CYP activity; Venkateswarlu K et al.; The disruption of Saccharomyces cerevisiae NADPH- cytochrome P450 oxidoreductase (CPR) gene resulted in a viable strain accumulating approximately 25% of the ergosterol observed in a sterol wild-type parent . The associated phenotypes could be reversed in transformants after expression of native CPR and a mutant lacking the N-terminal 33 amino acids, which localized in the cytosol . This indicated availability of the CPR in each case to function with the monooxygenases squalene epoxidase, CYP51, and CYP61 in the ergosterol biosynthesis pathway . Purification of the cytosolic mutant CPR indicated properties identical to native CPR and an ability to reconstitute ergosterol biosynthesis when added to a cell-free system, as well as to allow reconstitution of activity with purified CYP61, sterol 22-desaturase . This was also observed for purified Candida albicans and human CYP51 in reconstituted systems . The ability of the yeast enzyme to function in a soluble form differed from human CPR, which is shown to be inactive in reconstituting CYP activity. Fungal Genet Biol, 1997 Dec, 22(3), 199 - 208 Umchs5, a gene coding for a class IV chitin synthase in Ustilago maydis; Xoconostle-Cazares B et al.; A fragment corresponding to a conserved region of a fifth gene coding for chitin synthase in the plant pathogenic fungus Ustilago maydis was amplified by means of the polymerase chain reaction (PCR) . The amplified fragment was utilized as a probe for the identification of the whole gene in a genomic library of the fungus . The predicted gene product of Umchs5 has highest similarity with class IV chitin synthases encoded by the CHS3 genes from Saccharomyces cerevisiae and Candida albicans, chs-4 from Neurospora crassa, and chsE from Aspergillus nidulans . Umchs5 null mutants were constructed by substitution of most of the coding sequence with the hygromycin B resistance cassette . Mutants displayed significant reduction in growth rate, chitin content, and chitin synthase activity, specially in the mycelial form . Virulence to corn plantules was also reduced in the mutants . PCR was also used to obtain a fragment of a sixth chitin synthase, Umchs6 . It is suggested that multigenic control of chitin synthesis in U . maydis operates as a protection mechanism for fungal viability in which the loss of one activity is partially compensated by the remaining enzymes . J Exp Med, 1998 Feb 2, 187(3), 307 - 17 Endogenous interleukin 4 is required for development of protective CD4+ T helper type 1 cell responses to Candida albicans; Mencacci A et al.; Interleukin (IL)-4-deficient mice were used to assess susceptibility to systemic or gastrointestinal Candida albicans infections, as well as parameters of innate and elicited T helper immunity . In the early stage of systemic infection with virulent C . albicans, an unopposed interferon (IFN)-gamma response renders IL-4-deficient mice more resistant than wild-type mice to infection . Yet, IL-4-deficient mice failed to efficiently control infection in the late stage and succumbed to it . Defective IFN-gamma and IL-12 production, but not IL-12 responsiveness, was observed in IL-4-deficient mice that failed to mount protective T helper type 1 cell (Th1)-mediated acquired immunity in response to a live vaccine strain of the yeast or upon mucosal immunization in vivo . In vitro, IL-4 primed neutrophils for cytokine release, including IL-12 . However, late treatment with exogenous IL-4, while improving the outcome of infection, potentiated CD4(+) Th1 responses even in the absence of neutrophils . These findings indicate that endogenous IL-4 is required for the induction of CD4(+) Th1 protective antifungal responses, possibly through the combined activity on cells of the innate and adaptive immune systems. J Infect Dis, 1998 Mar, 177(3), 812 - 4 Histidine-based zinc-binding sequences and the antimicrobial activity of calprotectin; Loomans HJ et al.; Calprotectin is a protein in neutrophil cytoplasm and abscess fluids that appears to inhibit microbial growth through competition for zinc . This study was undertaken to identify specific sites that might be responsible for the protein's zinc-binding antimicrobial activity . A review of published calprotectin amino acid sequences revealed the HEXXH motif of thermolysin-type metalloproteases and an HHH polyhistidine sequence near the C-terminus of the protein's heavy chain . Reagent polyhistidine had antimicrobial activity against Candida albicans similar to that of calprotectin . Also, one type of HEXXH-containing thermolysin was inactive in the C . albicans assay, whereas a protein tagged with six C-terminal histidines did have calprotectin-like zinc-reversible antimicrobial activity . The activity of polyhistidine, as well as that of calprotectin itself, was reversed by addition of zinc or treatment with the histidine-modifying compound diethylpyrocarbonate . These results suggest that calprotectin's antimicrobial activity may be related to certain histidine-based zinc-binding sequences. New Microbiol, 1998 Jan, 21(1), 41 - 8 The prevalence of Candida albicans populations in the mouths of complete denture wearers; Abu-Elteen KH et al.; Using imprint cultures, the prevalence of oral candidosis and the frequency of isolation of Candida albicans and its density in oral mucosal sites and denture surfaces was determined in 190 healthy dentate subjects and 230 complete denture wearers . Candida colonization was 36.8% and 78.3% in healthy dentate and complete denture wearers patients, respectively . In the healthy dentate subjects the tongue, palate and cheeks, and in complete denture wearers additionally the upper and lower dentures, were the most frequently and densely colonized oral sites . Oral carrier rate and density of C . albicans were both higher in the denture wearers diabetic group than in the control non-diabetic group . Smoking was associated with an increase in the frequency and density of the yeast in denture wearers . Attention to these predisposing factors could reduce the incidence of oral candidosis particularly in immunocompromised patients. Eur J Med Res, 1997 Dec 31, 2(12), 507 - 13 Inhibitory effect of heparin on neutrophil phagocytosis and burst production using a new whole-blood cytofluorometric method for determination; Salih H et al.; The influence of heparin on Polymorphonuclear (PMN s) leukocytes was investigated using a new whole-blood cytofluorometric method (patent granted for the test with the number P 4334935.8-41) with Candida albicans and Staphylococcus aureus as test microorganisms . After comparing the effect of equal volumes of two widely used heparins we examined the influence of 5 different heparin-concentrations . Using both yeasts and bacteria, we found a significant, dose-depending decrease of the percentage of phagocyting PMN's and of phagocytized microorganisms as well as of the resulting percentage of PMN s producing respiratory burst along the kinetics . Furthermore we could demonstrate that heparin independently of phagocytosis produces a dose-dependent decrease of burst production of PMN's . Our results indicate that the use of heparins as anticoagulant for immunological investigations as well as clinically with patients under immunosuppressive therapy should be critically reconsidered . This applies even more because due to the evaluated dose-dependent decrease of phagocyte function no boundary for the inhibiting effect can be declared. Hepatogastroenterology, 1998 Jan-Feb, 45(19), 119 - 22 Gut colonization of mice by yeast: effects of methylprednisolone and antibiotics; Maraki S et al.; BACKGROUND/AIMS: This study evaluated the effects of broad spectrum antibiotics and methylprednisolone on the gut colonization of mice by C . albicans . METHODOLOGY: Male Crl:CD1 (ICR) BR mice, 3 months of age, were fed chow containing Candida albicans, while similar mice were fed regular chow . The gut of the Candida-fed mice was colonized by yeast . Groups of mice were subsequently treated for 10 days, with either ceftriaxone, ticarcillin-clavulanic acid, or methylprednisolone, each alone or with the combination of methylprednisolone and each antibiotic . Other Candida-colonized mice received normal saline, and non-colonized mice, serving as controls, received the same drugs and drug combinations or saline . RESULTS: Candida-colonized mice treated with each antibiotic alone had significantly higher yeast counts in their stool, while those treated with methylprednisolone alone did not . Colonized mice treated with the combination of each antibiotic with methylprednisolone had similar stool concentrations of Candida as mice treated with each antibiotic alone . Saline did not change Candida in the stool concentration . Yeast was not found in the stool of non-colonized mice treated with the drugs under investigation or saline . Dissemination of Candida did not occur in any mouse . CONCLUSIONS: Ceftriaxone and ticarcillin-clavulanic acid significantly increase gut colonization of mice by yeast, while methylprednisolone, either alone or in combination with these antibiotics, does not. Eur J Clin Microbiol Infect Dis, 1997 Dec, 16(12), 924 - 8 Evaluation of the susceptibility of pathogenic Candida species to fluconazole . Fluconazole Global Susceptibility Study Group; Bille J et al.; A fluconazole 25 microg disk diffusion test was used to test 2230 consecutively isolated Candida strains from 42 different hospital laboratories in 23 countries . Ninety seven percent of 1634 Candida albicans isolates and 83.4% of 596 non-Candida albicans isolates were susceptible to fluconazole, applying the proposed breakpoints (> or = 26 mm for susceptible strains and 18-25 mm for dose-dependent susceptible strains) . This is the first hospital laboratory study to evaluate a large number and wide range of sequential Candida isolates from patients with all types of hospital infections . The fluconazole disk diffusion test appears to be a low-cost, reproducible, and accurate means of assessing the in vitro susceptibility of Candida isolates. Pediatr Infect Dis J, 1998 Feb, 17(2), 130 - 4 Clustering of Candida infections in the neonatal intensive care unit: concurrent emergence of multiple strains simulating intermittent outbreaks; Khatib R et al.; BACKGROUND: Clusters of Candida albicans and Candida parapsilosis infections were noted intermittently in our neonatal intensive care unit (NICU) . We attempted to determine whether these clusters represented single strain outbreaks or coincidental emergence of unrelated strains . METHODS: A retrospective examination of the frequency of candidemia during a 9-year period, two point prevalence studies of colonization and assessment of strain relatedness of individual infant isolates during and in between clusters during a 2-year period with karyotyping and restriction endonuclease analysis of genomic DNA (REAG) . RESULTS: C . albicans and C . parapsilosis infections emerged in a scattered pattern (1 to 2 cases every few months) with intermittent clustering of 3 cases/month . The colonization rate was 50% 5 weeks after an apparent cluster, equally distributed between C . albicans and C . parapsilosis, and 17.6% (exclusively with C . parapsilosis) 4 months after absence of invasive disease . Utilizing REAG or karyotyping singly we noted 12 and 16 DNA banding patterns, respectively, among 23 infant isolates . Few patterns were observed repeatedly over 2- to 20-month periods, implying recurrent emergence of the same strains . Combining karyotyping with REAG revealed a different epidemiologic pattern . It identified 20 distinct composites with identical composites in 3 infant pairs . All infants with identical composites were in the NICU concurrently . The frequency of strain relatedness was comparable among clustered cases (16.7%), scattered cases (7.7%) and simultaneously colonized infants (16.7%) . CONCLUSIONS: These findings illustrate that Candida infections clustered periodically in our NICU but that these clusters were often caused by unrelated strains with infrequent cross-infection during and between clusters . With suboptimal typing this pattern of emergence can be mistaken for same strain outbreaks. Heart Lung, 1998 Jan-Feb, 27(1), 67 - 8 Penicillium peritonitis in a patient receiving continuous ambulatory peritoneal dialysis; Qadir MT et al.; Infection is a common complication in patients who receive continuous ambulatory peritoneal dialysis (CAPD) . Fungi causing peritonitis in these patients is less common compared with bacterial peritonitis . Fungal peritonitis accounts for less than 10% of cases in chronic CAPD, which usually follows either bacterial peritonitis or earlier exposure to broad-spectrum antibiotics . Most of these cases are caused by Candida albicans or other Candida species . There are only two case reports of Penicillium species peritonitis in patients with CAPD in the literature . We report the known third case of Penicillium species-related peritonitis in a patient receiving CAPD . The patient's condition improved dramatically after catheter removal. Pediatr Infect Dis J, 1998 Feb, 17(2), 146 - 8 Liposomal amphotericin B treatment for neonatal fungal infections; Scarcella A et al.; BACKGROUND: Disseminated fungal infections are a major problem in high risk neonates . Conventional antifungal agents are often unsatisfactory and have a high incidence of severe adverse effects . METHODS: We administered liposomal encapsulated amphotericin B (AmBisome), which is an alternative to conventional amphotericin B, to 40 preterm (mean birth weight, 1090 +/- 313.6 g; mean gestational age, 28.35 +/- 2.13 weeks) and 4 full term (mean birth weight, 3080 +/- 118 g; mean gestational age, 39 +/- 0.7 weeks) newborn infants with a severe fungal infection . RESULTS: Candida albicans was the most frequent fungus isolated (70%) . The duration of intravenous AmBisome therapy ranged from 7 to 49 days; the cumulative dose ranged from 7 to 138.8 mg/kg (median, 45.2 mg/kg) . Administration of AmBisome was effective in 72.7% of patients; 5 of 6 cases of meningitis also recovered; 63.6% of 33 very low birth weight infants survived . No side effects were observed . CONCLUSIONS: To our knowledge this is the largest study of the treatment of neonates with liposomal amphotericin B, and the results confirm its effectiveness and safety . However, randomized clinical trials are required to establish the most effective administration protocol for AmBisome, i.e . the starting dosage, the maximum effective dosage and the cumulative dosage, and to verify whether the preparation should be associated with another antifungal agent. Nihon Kyobu Shikkan Gakkai Zasshi, 1997 Nov, 35(11), 1271 - 7 {A patient with allergic bronchopulmonary candidiasis showing a high serum level of soluble interleukin 2 receptors}; Takabatake N et al.; An 84-year-old man was admitted to Yonezawa City Hospital with fever, cough, hemoptysis and progressive dyspnea . He had complained of wheezing asthmatoid and exertional dyspnea for the previous 10 years, regardless of the season . On admission, chest radiographs revealed a diffuse ground-glass shadow, fibrotic change, and volume reduction . Arterial blood gas analysis showed extreme hypoxemia . A computed tomographic (CT) scan of the chest showed not only faint ground-glass opacities and dense patches in the whole lung field, but also central bronchiectasis . Laboratory tests revealed that both total serum levels of IgE and specific IgE for Candida albicans were elevated . In the bronchoalveolar lavage fluid, lymphocyte, neutrophil and eosinophil percentages were high, and the CD4/CD8 ratio was low . We diagnosed the fibrotic stage of allergic bronchopulmonary candidiasis . During treatment with hydrocortisone and fluconazole, eosinophilia in the peripheral blood was observed, and serum candida antigen was positive . In addition, high serum levels of soluble interleukin 2 receptors were observed in this patient. Microbiology, 1998 Feb, 144 ( Pt 2), 425 - 32 Isolation of CaSLN1 and CaNIK1, the genes for osmosensing histidine kinase homologues, from the pathogenic fungus Candida albicans; Nagahashi S et al.; Recent studies have revealed that fungi possess a mechanism similar to bacterial two-component systems to respond to extracellular changes in osmolarity . In Saccharomyces cerevisiae, Sln1p contains both histidine kinase and receiver (response regulator) domains and acts as an osmosensor protein that regulates the downstream HOG1 MAP kinase cascade . SLN1 of Candida albicans was functionally cloned using an S . cerevisiae strain in which SLN1 expression was conditionally suppressed . Deletion analysis of the cloned gene demonstrated that the receiver domain of C . albicans Sln1p was not necessary to rescue SLN1-deficient S . cerevisiae strains . Unlike S . cerevisiae, a null mutation of C . albicans SLN1 was viable under regular and high osmotic conditions, but it caused a slight growth retardation at high osmolarity . Southern blotting with C . albicans SLN1 revealed the presence of related genes, one of which is highly homologous to the NIK1 gene of Neurospora crassa . Thus, C . albicans harbours both SLN1- and NIK1-type histidine kinases. Microbiology, 1998 Feb, 144 ( Pt 2), 411 - 24 A role for the MAP kinase gene MKC1 in cell wall construction and morphological transitions in Candida albicans; Navarro-Garcia F et al.; The Candida albicans MKC1 gene encodes a mitogen-activated protein (MAP) kinase, which has been cloned by complementation of the lytic phenotype associated with Saccharomyces cerevisiae slt2 (mpk1) mutants . In this work, the physiological role of this MAP kinase in the pathogenic fungus C . albicans was characterized and a role for MKC1 in the biogenesis of the cell wall suggested based on the following criteria . First, C . albicans mkc1 delta/mkc1 delta strains displayed alterations in their cell surfaces under specific conditions as evidenced by scanning electron microscopy . Second, an increase in specific cell wall epitopes (O-glycosylated mannoprotein) was shown by confocal microscopy in mkc1 delta/mkc1 delta mutants . Third, the sensitivity to antifungals which inhibit (1,3)-beta-glucan and chitin synthesis was increased in these mutants . In addition, evidence for a role for the MKC1 gene in morphological transitions in C . albicans is presented based on the impairment of pseudohyphal formation of mkc1 delta/mkc1 delta strains on Spider medium and on the effect of its overexpression on Sacch . cerevisiae colony morphology on SLADH medium . Using the two-hybrid system, it was also demonstrated that MKC1 is able to interact specifically with Sacch . cerevisiae Mkk1p and Mkk2p, the MAP-kinase kinases of the PKC1-mediated route of Sacch . cerevisiae, and to activate transcription in Sacch . cerevisiae when bound to a DNA-binding element . These results suggest a role for this MAP kinase in the construction of the cell wall of C . albicans and indicate its potential relevance for the development of novel antifungals. Microbiology, 1998 Feb, 144 ( Pt 2), 391 - 401 Regulation of chitin synthesis during dimorphic growth of Candida albicans; Munro CA et al.; Candida albicans has three genes encoding chitin synthase enzymes . In wild-type strains, the expression of CHS2 and CHS3 peaked 1-2 h after the induction of hyphal growth, whilst mRNA levels in a non-germinative strain, CA2, remained low under the same conditions . CHS1 gene expression did not peak during germ tube formation but remained at low levels in both yeast and hyphal growth . The pattern of gene expression did not predict the changes in measured chitin synthase activities or changes in chitin content during dimorphic transition . Chitin synthase activity increased steadily, and did not peak shortly after germ tube induction, and activity profiles were similar in germ-tube-competent and germ-tube-negative strains . The phenotype of a delta chs2 null mutant suggested that CHS2 encoded the major enzyme activity in vitro and was largely responsible for elevated chitin synthase activities in microsomal preparations from hyphal cells compared to yeast cells . However, CaChs3p was responsible for synthesis of most chitin in both yeast and hyphae . Three independent chitin assays gave markedly different estimates of the relative chitin content of yeast and hyphae and wild-type and chs mutants . Only one of the methods gave a significantly higher chitin content for hyphal compared to yeast cell walls and a lower chitin content for hyphae of the delta chs2 null mutant compared to the parental strain. Mol Microbiol, 1998 Feb, 27(3), 587 - 98 A Ste6p/P-glycoprotein homologue from the asexual yeast Candida albicans transports the a-factor mating pheromone in Saccharomyces cerevisiae; Raymond M et al.; In Saccharomyces cerevisiae MATa cells, export of the a-factor mating pheromone is mediated by Ste6p, a member of the ATP-binding cassette (ABC) superfamily of transporters and a close homologue of mammalian multidrug transporter P-glycoproteins (Pgps) . We have used functional complementation of a ste6delta mutation to isolate a gene encoding an ABC transporter capable of a-factor export from the pathogenic yeast, Candida albicans . This gene codes for a 1323-amino acid protein with an intramolecular duplicated structure, each repeated half containing six potential hydrophobic transmembrane segments and a hydrophilic domain with consensus sequences for an ATP-binding fold . The predicted protein displays significant sequence similarity to S . cerevisiae Ste6p and mammalian Pgps . The gene has been named HST6, for homologue of STE6 . A high degree of structural conservation between the STE6 and the HST6 loci with respect to DNA sequence, physical linkage and transcriptional arrangement indicates that HST6 is the C . albicans orthologue of the S . cerevisiae STE6 gene . We show that the HST6 gene is transcribed in a haploid-specific manner in S . cerevisiae, consistent with the presence in its promoter of a consensus sequence for Mata1p-Matalpha2p binding known to mediate the repression of haploid-specific genes in S . cerevisiae diploid cells . In C . albicans, HST6 is expressed constitutively at high levels in the different cell types analysed (yeast, hyphae, white and opaque), demonstrating that HST6 transcription is not repressed in this diploid yeast, unlike in diploid S . cerevisiae, and suggesting a basic biological function for the Hst6p transporter in C . albicans . The strong similarity between Hst6p and the multidrug transporter Pgps also raises the possibility that Hst6p could be involved in resistance to antifungal drugs in C . albicans. Infect Immun, 1998 Mar, 66(3), 1273 - 5 Acute neutropenia decreases inflammation associated with murine vaginal candidiasis but has no effect on the course of infection; Black CA et al.; We have used a mouse model of vaginal candidiasis to determine the effect of neutrophil depletion on (a) the clearance of Candida albicans and (b) the degree of inflammation associated with infection . No differences in recoverable yeast number or rate of clearance were observed between normal and neutrophil-depleted mice; however, vaginal inflammation was significantly decreased in neutrophil-depleted animals. Infect Immun, 1998 Mar, 66(3), 966 - 73 MP1 encodes an abundant and highly antigenic cell wall mannoprotein in the pathogenic fungus Penicillium marneffei; Cao L et al.; We cloned the MP1 gene, which encodes an abundant antigenic cell wall mannoprotein from the dimorphic pathogenic fungus Penicillium marneffei . MP1 is a unique gene without homologs in sequence databases . It codes for a protein, Mp1p, of 462 amino acid residues, with a few sequence features that are present in several cell wall proteins of Saccharomyces cerevisiae and Candida albicans . It contains two putative N glycosylation sites, a serine- and threonine-rich region for O glycosylation, a signal peptide, and a putative glycosylphosphatidylinositol attachment signal sequence . Specific anti-Mp1p antibody was generated with recombinant Mp1p protein purified from Escherichia coli to allow further characterization of Mp1p . Western blot analysis with anti-Mp1p antibody revealed that Mp1p has predominant bands with molecular masses of 58 and 90 kDa and that it belongs to a group of cell wall proteins that can be readily removed from yeast cell surfaces by glucanase digestion . In addition, Mp1p is an abundant yeast glycoprotein and has high affinity for concanavalin A, a characteristic indicative of a mannoprotein . Furthermore, ultrastructural analysis with immunogold staining indicated that Mp1p is present in the cell walls of the yeast, hyphae, and conidia of P . marneffei . Finally, it was observed that infected patients develop a specific antibody response against Mp1p, suggesting that this protein represents a good cell surface target for host humoral immunity. Shock, 1998 Feb, 9(2), 146 - 52 Autocrine/paracrine modulation of polymorphonuclear leukocyte survival after exposure to Candida albicans; Sweeney JF et al.; Polymorphonuclear leukocytes (PMN) play a central role in the host response to injury and infection . These terminally differentiated phagocytes have a limited life span, after which they undergo spontaneous apoptosis . PMN life span can be significantly prolonged by several naturally occurring cytokines, and PMN are now known to be capable of cytokine production in response to various antigenic stimuli . These facts suggest the possibility that PMN possess an autocrine/paracrine mechanism for the control of their own survival . The present study was undertaken to test this hypothesis . Supernatants from PMN that had been incubated with Candida albicans for 18 h significantly decreased the number of fresh PMN demonstrating features of apoptosis and increased the percentage of viable PMN during in vitro culture . This was demonstrated by monitoring morphologic features of apoptosis with fluorescence microscopy and DNA endonuclease activity with agarose gel electrophoresis . Significant levels of tumor necrosis factor (TNF) were detectable in the supernatants of PMN that had been stimulated with C . albicans, as determined using a TNF-sensitive cell line . Neutralization of TNF biologic activity with a specific monoclonal antibody partially abrogated the supernatant-mediated prolongation of PMN survival . The present study demonstrates that PMN possess a mechanism for the modulation of their own survival, which in part may be through the production of TNF. Int Immunol, 1998 Jan, 10(1), 37 - 48 Defective co-stimulation and impaired Th1 development in tumor necrosis factor/lymphotoxin-alpha double-deficient mice infected with Candida albicans; Mencacci A et al.; To define the immunological functions of tumor necrosis factor (TNF) in Candida albicans infection, TNF/lymphotoxin (LT)-alpha double-deficient mice were assessed for susceptibility to systemic or gastrointestinal infection and parameters of innate and adaptive Th immunity . When compared to wild-type mice, TNF/LT-alpha-deficient mice were more susceptible to either type of infection caused by virulent or low-virulence C . albicans cells . Susceptibility to infection correlated with impaired development of protective Th1 responses, in spite of the production of bioactive IL-12 . The occurrence of predominant Th2 responses was associated with both impaired antifungal effector functions of neutrophils and a defective expression of co-stimulatory molecules on macrophages . All functions were improved upon administration of recombinant TNF-alpha, also resulting in increased resistance to infection . These findings indicate that the protective effect of TNF-alpha in candidiasis relies on the induction of antifungal Th1 responses, possibly occurring through stimulation of antifungal effector functions and co-stimulatory activities of phagocytic cells. East Afr Med J, 1997 Jun, 74(6), 389 - 91 Adherence of Candida albicans to human vaginal epithelial cells; Nwobu RA et al.; The adherence capacity of Candida species to female vaginal epithelial cells was examined . The results showed that in four groups of patients studied, the highest adherence was with epithelial cells collected from pregnant diabetic women (47% adherence, and 1,700 adherent yeasts) . Pregnant or diabetic women had 39% each of adherence to epithelial cells but differed in the number of adherent yeasts (1,400 and 1,000 respectively) . The diabetic and pregnant women therefore appeared differential attachment to epithelial cells from different physiologically adapted women. FEMS Microbiol Lett, 1998 Feb 1, 159(1), 129 - 35 Altered adherence in strains of Candida albicans harbouring null mutations in secreted aspartic proteinase genes; Watts HJ et al.; The aspartate proteinase inhibitor pepstatin A has been shown previously to reduce the adherence of Candida albicans yeast cells to human surfaces . This suggests that in addition to their presumed function facilitating tissue penetration, the secreted aspartate proteinases (Saps) of this fungal pathogen may have auxiliary roles as cellular adhesins . We therefore examined the relative adherence of yeast cells of a parental wild-type strain of C . albicans in relation to yeast cells of strains harbouring specific disruptions in various members of the SAP gene family in an otherwise isogenic background . The adhesiveness of delta sap1, delta sap2, delta sap3 null mutants and a triple delta sap 4-6 disruptant was examined on three surfaces--glass coated with poly-L-lysine or a commercial cell-free basement membrane preparation (Matrigel) and on human buccal epithelial cells . Pepstatin A reduced adherence to all surfaces . Adherence of the each of the single SAP null mutants to these three substrates was either reduced or not affected significantly compared to that of the parental strain . The adherence of the delta sap4-6 mutant was reduced on poly-L-lysine and Matrigel, but increased on buccal cells . The results suggest that in addition to a primary enzymatic role, various SAPs may also act singly or synergistically to enhance the adhesiveness to C . albicans cells to certain human tissues. Ter Arkh, 1997, 69(11), 41 - 4 {Candida albicans sensitization in patients with atopic bronchial asthma and atopic dermatitis}; Samuilova TL et al.; Candida albicans, a component of normal human microflora, can induce synthesis of specific IgE-antibodies in patients with atopic bronchial asthma and atopic dermatitis . The study included 25 patients with atopic dermatitis sensitized to C.albicans and 23 patients with atopic dermatitis non-sensitized to C.albicans . The sensitization was determined by the skin test and enzyme immunoassay . The patients had the history of atopic dermatitis exacerbation after taking food containing baking yeasts . Atopic dermatitis with sensitization to C.albicans is characterized by severe course correlating with the following indices: high total IgE (r = 0.6), level of IgE antibodies to C.albicans (r = 0.6), level of serum IgG (r = 0.46) and IgA (r = 0.33) . Contrary to adults, children with sensitization to C.albicans had decreased relative number of CD4+, CD8+ and CD72+ of lymphocyte subpopulations . Thus, sensitization to C.albicans manifests in severe atopic dermatitis which in children is often associated with immune deficiency. Curr Biol, 1997 Nov 1, 7(11), R691 - 4 Candida pathogenesis: unravelling the threads of infection; Corner BE et al.; Recent studies are beginning to delineate those pathways by which the important pathogen Candida albicans switches from one growth form to another; at the same time, insights are being gained into the importance of growth form in pathogenesis. Mycoses, 1997 Nov, 40(7-8), 283 - 9 Response to fluconazole and itraconazole of Candida spp . in denture stomatitis; Martin-Mazuelos E et al.; The significance of Candida albicans in the development of denture stomatitis (DS), as well as the clinical and microbiological efficacy of treatment with fluconazole and itraconazole was studied in 115 patients affected with DS and 200 controls (100 healthy patients with dental prosthesis and 100 healthy patients without prosthesis) . Specimens were taken from all patients; subsequently all patients with positive culture of the DS group were treated with fluconazole . A second specimen was taken after 15 days of treatment with fluconazole, and if the results were positive again, treatment with itraconazole was instituted and the patients were given appointments for taking a third specimen . The incidence of C . albicans was 92% in the group of patients with DS . After treatment with fluconazole, a clinical cure of 97% and a microbiological cure of 78% was obtained in the patients with DS . In 3.2% of the cases strains resistant to fluconazole were found . The cases of microbiological resistance to fluconazole were treated with itraconazole resulting in a clinical cure of 100% and a microbiological cure of 77% . The results show the poor correlation of the clinico-microbiological response after treatment with these antifungal agents in denture stomatitis. Mycoses, 1997 Nov, 40(7-8), 279 - 82 Fluconazole susceptibility of Candida isolates from oropharyngeal candidosis; Dannaoui E et al.; Fifty strains of Candida isolated from 38 patients with oropharyngeal candidosis were tested in vitro for fluconazole susceptibility with a disk diffusion test and for determination of minimal inhibitory concentrations (MICs) . For 25 patients treated with fluconazole, the relationship between in vitro susceptibility and clinical outcome was analysed . A good correlation between in vitro results and therapeutic efficacy was found . In only one case was treatment failure associated with a susceptible strain . Mixed cultures of Candida albicans and non-albicans Candida species were not uncommon and, more interestingly, some samples contained different strains of C . albicans with varied fluconazole susceptibilities . Good agreement was observed between the two techniques used for fluconazole susceptibility testing. Mycoses, 1997 Nov, 40(7-8), 255 - 8 Typing of Candida albicans by use of a combined PCR/hybridization assay; Vogeser M et al.; A modified genotyping assay for Candida albicans based on the RAPD-PCR (random amplification of polymorphic DNA) is described . Following capillary blot of RAPD-PCR products onto a nylon membrane, hybridization with a PCR-generated gene probe resulted in heterogeneous band patterns . By this method the discriminatory power of RAPD-PCR is enhanced with satisfactory reproducibility and reduced interference. Mycoses, 1997 Nov, 40(7-8), 249 - 53 A method for recovery of Candida albicans DNA from larger blood samples and its detection by polymerase chain reaction on proteinase genes; Reichard U et al.; A method for the detection of Candida albicans from up to 15 ml of blood by polymerase chain reaction (PCR), based on the differential resistance of mammalian and fungal cells towards detergent was developed . The procedure essentially involved removal of the blood cells by sodium dodecyl sulfate (SDS) induced lysis, followed by DNA extraction after degradation of fungal cell walls by a recombinant beta-1,3-glucanase . The genes of two different aspartic proteinases from C . albicans, SAP1 and SAP2, with an overall homology of 77% in their nucleotide sequences, were chosen as targets for PCR . The oligonucleotide primers used were directed to strictly conserved regions similar in both genes . As the number of base pairs between the primers are different in the two genes, amplification products of 220 bp and 238 bp in length were obtained . This led to a characteristic double band in subsequent agarose gel electrophoresis . The detection limit for a nested PCR was less than 10 C . albicans cells ml-1 of seeded blood . The detection limit of conventional PCR from a blood volume in the 10 ml range was less than 100 yeasts ml-1 . Preliminary trials with clinical blood specimens suggested, that conventional PCR from large blood samples, being less laborious and prone to contamination than nested PCR, could be suited for the detection of deepseated C . albicans mycosis. Mycoses, 1997, 40 Suppl 2, 9 - 12 Switch of phenotype as an escape mechanism of the intruder; Odds EC; Phenotypic switching in Candida albicans is a reversible, high-frequency phenomenon that is readily detectable in a fungal population as changes in cell or colony morphology . Some putative attributes of virulence in C . albicans, including expression of cell wall glycoproteins, secretion of proteolytic enzymes and hypha formation have been associated with switching phenomena . C . albicans isolates from active infection tend to show a higher prevalence of phenotypic switching than those associated with commensalism . Moreover, some characteristics of azole resistance in C . albicans are compatible with a switch of phenotype . There is thus a preliminary basis of scientific evidence for a hypothesis that phenotypic switching may indeed serve as an attribute of virulence in at least one pathogenic fungus, facilitating invasion and escape from host defences. Mycoses, 1997 Oct, 40(5-6), 187 - 92 Interactions between amphotericin B and nitroimidazoles against Candida albicans; Cury AE et al.; This work proved that nitroimidazole antiprotozoal agents, such as metronidazole, ornidazole, secnidazole and tinidazole, in concentrations of up to 64 micrograms ml-1 did not present any antifungal activity against 17 strains of Candida albicans . The combination of each drug with amphotericin B showed the occurrence of variable interactions according to the studied strain . Promising results were observed based on synergistic and additive interactions of the polyene with the metronidazole; the inhibitory and lethal activities of the drugs were potentiated against all strains in concentrations reachable in vivo. Mycoses, 1997 Oct, 40(5-6), 179 - 85 Prevalence and susceptibility of vaginal yeast isolates in Jordan; Abu-Elteen KH et al.; The prevalence of vaginal yeast species has been studied in 140 women (41 pregnant, 66 infertile and 33 healthy controls) attending a gynaecological private clinic in Amman, Jordan . Yeast species were isolated from pregnant (68.2%), infertile (51.5%) and healthy control (48.4%) women . Patients manifesting one, two or three symptoms of vulvovaginitis were 22.1%, 26.8% or 24.2% respectively . Asymptomatic cases and cases with more than three symptoms were 22.4% and 4.5% respectively . Candida albicans was the dominant species (in 51.3% of the patients) followed by C . glabrata (17.9%) . The percentage occurrence as well as the pattern of Candida species differed among the different groups of patients . Candida kefyr was found to be significantly higher in the infertile women . In vitro sensitivity tests using amphotericin B, nystatin, miconazole nitrate and chlorhexidine were carried out; amphotericin B was the most effective and miconazole nitrate the least. Mycoses, 1997 Oct, 40(5-6), 169 - 73 Epidemiological study of Candida spp . colonization in cardiovascular surgical patients; Tran LT et al.; Candida infections involve multiple risk factors . Among the independent risk factors identified, the degree of colonization of Candida spp . allows the prediction of subsequent severe candidosis in surgical patients . The aim of this study was to assess among 13 selected variables, those that would best predict the perioperative variation of the colonization index (CI) of Candida spp . in cardiovascular surgical patients . The colonization index took into account the number of sites colonized and the density of growth . The results showed that 56.8% of our patients were colonized perioperatively . A total of 116 isolates were identified and Candida albicans accounted for 76.7% of the strains . Among the patients who developed post-surgical Candida infections, 57.1% had an increase of the CI early after the operation . By univariate analysis, three factors were significantly associated with an increase of the CI in patients after surgery; sex (female), the duration of central intravascular catheterization and the length of stay in the surgical intensive care unit (SICU) . Epidemiological data could help predict those patients who are at risk of developing Candida infections. Mycoses, 1997 Oct, 40(5-6), 159 - 67 Multilocus enzyme electrophoresis analysis of Candida albicans isolates from three intensive care units . An epidemiological study; Arnavielhe S et al.; To evaluate the mechanism and risk factor associated with the nosocomial acquisition of Candida albicans, a 3-month prospective study was conducted on non-neutropenic patients in three distinct intensive care units in distinct hospitals . A total of 43 samples from 19 patients has been typed by multilocus enzyme electrophoresis (MEE) . Samples (24) from the deep pharynx of hospital staff members were also cultured and typed . Thirteen of the 19 enzyme loci studied were polymorphic . The 52 electrophoretic types were assigned to 67 isolates, each type was represented by one to five isolates; this proved the great diversity of the isolates . It appeared that most patients were colonized or infected with different C . albicans strains . This study pointed out a possible cross-infection between patients and hospital staff and between healthy members of the hospital staff, and also showed the successive stages of infection by different electrophoretic types . Intubation and respiratory equipment as surgery intervention were possible sources of observed infections. Mycoses, 1997 Oct, 40(5-6), 153 - 7 Western blot analysis of the immune response to Candida albicans antigens in 391 long-term intensive care patients; Weis C et al.; The aim of this study was to determine, using Western blot, the prevalence of anti-Candida albicans antibodies in long-term intensive care patients and to characterize specific immune responses that may only occur in patients with invasive candidosis . A total of 1751 serum samples from 391 patients of a German multicentre study, which was designed to determine the incidence of systemic candidosis, was examined . Significantly enhanced antibody production against specific antigens was observed in several subgroups of patients, i.e . those with underlying disease of the pancreas (29 kDa, P = 0.006), cholecystolithiasis (47 kDa, P = 0.029), gastrointestinal tract disease (47 kDa, P = 0.03), steroid therapy (58 kDa, P = 0.02), thrush (58 kDa, P = 0.032), urogenital infection (58 kDa, P = 0.034), Candida antigen titre > or = 1:4 (58 kDa, P = 0.002) and positive fungal culture (36 kDa, P = 0.033) and those who had died (36 kDa, P = 0.011) . In contrast to earlier publications, an immune response against the 29 and 47 kDa antigens was relatively common among long-term intensive care patients (37% and 70% antibody positive respectively) . A single antigen that provided satisfactory sensitivity and specificity for the discrimination between fungal infection and no fungal infection or between superficial and invasive fungal infection was not identified in this study. Recenti Prog Med, 1997 Oct, 88(10), 479 - 84 {Mycotic vulvovaginitis}; Margariti PA et al.; Vulvovaginitis is the most common clinical manifestation of fungal infections causing human mycoses; the incidence occurs in 10% of women, during pregnancy the incidence achieves 30% of cases . Candida albicans has resulted to be the most commonly isolated agent in patients with fungemia . In fact, Candida appears to be the species recovered in as many as 90% of cases . They are mainly the sexual activity, hormonal contraception and several pathologies such as diabetes mellitus and thyroiditis responsible for the pathogenesis of infection . The first symptom of this infection is usually pruritus associated to leukorrhea, dyspareunia and vulvovaginal irritation . Antifungal therapy may be required in more severe cases of vulvovaginal candidiasis . Candida species can be identified on isolation culture media including agar and on direct examination . Diagnosis can also be made through san immunologic examination . However, the authors confirm that the risk factors together with a correct diagnosis of the Candida etiological agent in the different species (albicans, glabrata, tropicalis, krusei) should be accurately investigated in order to give the correct therapeutical approach. J Dent Hyg, 1996 Jul-Aug, 70(4), 161 - 5 An in vitro investigation of the efficacy of CPC for use in toothbrush decontamination; Meier S et al.; PURPOSE: A product designed as a toothbrush disinfectant containing cetylpyridinium chloride (CPC), a quaternary ammonium compound, recently was introduced . The purpose of this study was to provide additional evidence that CPC provides a practical solution for destroying residual microorganisms on air-dried toothbrushes and toothbrushes stored in a travel container . METHODS: Sterile synthetic toothbrushes were inoculated with optical density standardized laboratory cultures of Staphylococcus epidermidis or Candida albicans . Half were then disinfected with CPC and half were used as untreated controls . The toothbrushes were vortexed in sterile saline solution, diluted in a ten-fold series, and plated on 5% blood agar or Sabouraud dextrose agar . The plates were incubated at 37 degrees C in a normal atmosphere for 48 hours, and colonies were counted . RESULTS: CPC produced significant decreases in residual microorganisms . Using the CPC spray treatment on air-dried toothbrushes, Staphylococcus epidermidis essentially was reduced 100-fold, while Candida albicans had a 94% reduction of growth . Bacterial counts were higher in the samples stored in closed containers as compared to the air-dried samples . CONCLUSION: CPC appeared to be an effective toothbrush disinfectant for the organisms evaluated . It is practical and economical . CPC could easily fit into the recommendations of a practice committed to infection control. Mycoses, 1997 Dec, 40(9-10), 377 - 80 Quantitative cultures of Candida from mouthwash fluid in HIV-infected patients: a longitudinal study; Bergbrant IM et al.; The density of Candida colonization in mouthwash fluid of 59 HIV-seropositive patients and 21 controls was determined . No significant difference in colony-forming units was found . No correlation was found between the colonizing density of Candida albicans and the CD4 count among the patients . Twenty-seven of the HIV-seropositive patients were followed for almost 3 years . No difference was found between the number of Candida albicans colony-forming units at the first and second time of sampling . Vaccination with HIV IIIB GP 160 vaccine did not have any influence on the prevalence of Candida albicans. Mycoses, 1997 Dec, 40(9-10), 373 - 5 Candida albicans suppresses transcription of melanogenesis enzymes in cultured melanocytes; Kippenberger S et al.; Human skin can be colonized by different yeasts that may have an impact on skin pigmentation . In order to study this effect normal human melanocytes were cultured with different yeasts . Reverse transcription polymerase chain reaction (RT-PCR) analysis gives evidence that Candida albicans suppresses the transcription of melanogenesis enzymes. Mycoses, 1997 Dec, 40(9-10), 369 - 72 Standardized molecular typing of Candida albicans strains; Lischewski A et al.; A method is presented for the standardization of Candida albicans DNA fingerprinting, which is based on Southern hybridization of EcoRI-digested chromosomal DNA with the moderately repetitive DNA element CARE-2 and the subsequent rehybridization of the blots with a molecular size marker also included in each DNA sample . This method resulted in extremely precise alignment of all strain-specific CARE-2 hybridization patterns, even when analysed on different gels, and will enhance the accuracy of genetic relationship determinations in epidemiological studies including large numbers of strains. Mycoses, 1997 Dec, 40(9-10), 363 - 8 Adherence on buccal epithelial cells and germ tube formation in the continuous flow culture of clinical Candida albicans isolates; Wellmer A et al.; Mucosal adherence and germ tube formation have been considered as important virulence factors of Candida albicans . We investigated 11 clinical isolates (among them six isolates from oesophageal thrush) for quantification of adherence to buccal epithelial cells and germ tube formation in the continuous flow culture in vitro, and correlated the results with the clinical data of the patients . Adherence varied considerably between the different C . albicans strains . Strains recovered from clinically, culturally and serologically confirmed oesophageal thrush adhered stronger to buccal epithelial cells . Isolates from cases with heavy colonisation but clinically without candidosis were less adherent . Only after 30 min germ tube formation was observed in the continuous flow culture . Strains with stronger adherence also showed significantly faster and increased germ tube formation . The patients with oesophageal thrush did not suffer any particular immunosuppression such as HIV infection, although in most cases chronic alcoholism was apparent . We conclude, that in cases with minor immunosuppression the expression of the virulence factors adherence and germ tube formation plays an important role in the pathogenesis of candidosis, whereas it may be of less importance in cases with severe immunosuppression . In the latter they may, however, influence outcome. Mycoses, 1997 Dec, 40(11-12), 451 - 3 The formation of hyphae of Candida albicans induced by cyclodextrins; Fekete-Forgacs K et al.; Hyphal growth of Candida albicans was observed when yeast was cultured at 27 degrees C in liquid media containing 1% Tripcasine and 1.8% cyclodextrins (alpha, beta, and gamma respectively) . Tripcasine as the sole nitrogen source did not induce the formation of hyphae of C . albicans, but cyclodextrins, especially CD-beta, were able to induce yeast-mycelial transition . In the TCD-beta media 25-30% septate hyphae form was observed . This study indicates the existence of an uptake system for CDs in C . albicans, provided these compounds are linearised in the medium . The CDs are inducers of hyphae and they may enter the C . albicans cells as linear oligosaccharids. Mycoses, 1997 Dec, 40(11-12), 429 - 37 A synergistic effect of Carica papaya latex sap and fluconazole on Candida albicans growth; Giordani R et al.; A mixture of Carica papaya latex (0.41 mg protein ml-1) and fluconazole (2 micrograms ml-1) showed a synergistic action on the inhibition of Candida albicans growth . Thus, with