Candida albicans

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 this mixture an equivalent inhibition rate was observed to that obtained when C . albicans was cultured in a medium supplemented with a two-fold concentration (4 micrograms ml-1) of fluconazole alone . This synergistic effect resulted in partial cell wall degradation as indicated by transmission electron microscopy observations . An increase of fluconazole concentration from 2 micrograms ml-1 to 4 micrograms ml-1 involved a small decrease of MIC 80% from latex (150 to 130 micrograms protein ml-1) . Measure of MIC 80% from fluconazole mixed with latex in a subinhibitory concentration (85 micrograms protein ml-1) allows the determination of an effective fluconazole concentration (4 micrograms ml-1) inferior to mean plasmatic dose observed in human therapy . The potential therapeutic use of latex in combination with a synthetic antifungal is discussed.

Arch Microbiol, 1997 Dec, 168(6), 464 - 72
Regulation of N-acetylglucosaminidase production in Candida albicans; Niimi K et al.; The N-acetylglucosaminidase of Candida albicans is a secreted hydrolytic enzyme that contributes to the yeast's virulence . There was a significant increase in the N-acetylglucosaminidase activity of C . albicans cells released from carbon starvation in medium containing N-acetylglucosamine . The increased enzyme activity in N-acetylglucosamine-grown cells correlated with increased transcription of the HEX1 gene, which encodes C . albicans N-acetylglucosaminidase . In contrast, glucose repressed HEX1 transcription, and glucose-grown cells had on average 94-fold lower N-acetylglucosaminidase activities than did N-acetylglucosamine-grown cells . N-acetylglucosaminidase induction in cells grown on N-acetylglucosamine was also repressed by fructose, mannose or galactose, although to a lesser extent than by glucose, and sucrose repressed enzyme production by only 10% . Eighty-eight percent of the enzyme in N-acetylglucosamine-grown cells was localised in the periplasm, and after incubation for 5 h, 30 or 70% of the total enzyme activity was secreted into the medium by yeast or mycelial cells, respectively . The cellular location of the enzyme and the regulation of production by the carbon source indicate a scavenging role for C . albicans N-acetylglucosaminidase.

Early Hum Dev, 1997 Nov 24, 50(1), 61 - 70
The influence of septicaemia on spontaneous motility in preterm infants; Bos AF et al.; The qualitative assessment of general movements (GMs) in preterm infants is a sensitive method to investigate the integrity of the central nervous system . The question arises whether systemic infections affect the quality of GMs in a similar fashion to brain lesions . We were able to provide an answer to this problem in six infants (gestational age 24.4-32.4 weeks, birth weight 600-1660 grams), who had initially normal GMs as analyzed from sequential video-recordings . All infants sustained a proven septicaemia (Candida albicans in two, Staphylococcus aureus in three, a coagulase-negative staphylococcus in one infant) . Unintentionally, recordings were also made during the acute phase . The complexity and variability of the GMs remained largely intact in five of the six infants; only one infant had transiently abnormal GMs . Compared with 1 week before the acute phase, the speed and amplitude of the GMs were diminished, giving the GMs a sluggish appearance . One to two weeks after the acute phase of septicaemia, the quality of GMs, i.e . speed and amplitude, had normalized in all infants . This study demonstrates that it is possible to discriminate between abnormal GMs due to cerebral lesions and sluggish GMs due to severe systemic infections, when the complexity of the GMs is considered as the main characteristic for judgement of normality of GM-quality.

Oral Microbiol Immunol, 1997 Jun, 12(3), 168 - 73
Coaggregation of Candida albicans with oral Fusobacterium species; Grimaudo NJ et al.; Nine strains of oral Fusobacterium were examined for their ability to coaggregate in vitro with four strains of the oral yeast . Candida albicans . All of the Fusobacterium nucleatum strains and Fusobacterium periodontium and Fusobacterium sulci coaggregated to various degrees with all of the Candida strains . Fusobacterium alocis, Fusobacterium mortiferum and Fusobactrium simiae strains did not coaggregate with any of the Candida strains . Exposure of the coaggregating Fusobacterium strains but not the Candida strains to heat, trypsin, and proteinase K eliminated coaggregation . Amphotericin B or trichodermin treatment of the yeast species had no effect . The reactions were inhibited by addition of 0.1 M mannose, glucosamine and alpha-methyl mannoside . All coaggregating pairs were disaggregated by the addition of sodium dodecyl sulfate, but nonionic detergents had no effect . The addition of 2.0 M urea completely reversed coaggregation . Candida strains were sensitive to periodate oxidation, whereas the Fusobacterium strains were stable to this treatment . All coaggregations occurred in the presence of saliva and appeared stronger than in buffer . These data suggest that the coaggregations involve either a protein or glycoprotein on the Fusobacterium surface, which may interact with carbohydrates or carbohydrate-containing molecules on the surface of the Candida . These observations expand the known range of intergeneric coaggregations occurring between human oral microbes and indicate that coaggregation of C . albicans and Fusobacterium species may be an important factor in oral colonization by this yeast . The authors believe this to be the first description of coaggregation concerning a carbohydrate component on the yeast cell and a protein component on the oral bacterial cell.

J Clin Microbiol, 1998 Feb, 36(2), 395 - 401
Rapid detection of Candida albicans in clinical samples by DNA amplification of common regions from C . albicans-secreted aspartic proteinase genes; Flahaut M et al.; Laboratory diagnosis based on genomic amplification methods such as PCR may provide an alternative and more sensitive method than conventional culture for the early detection of deep-seated candidiasis, an increasing cause of morbidity and mortality among immunocompromised patients . A novel method of DNA extraction from clinical samples based on treatment with proteinase K and isolation of DNA on a silica membrane was developed . The targets used for DNA amplification were the Candida albicans-secreted aspartic proteinase (SAP) genes, a multiple-gene family of at least seven members in C . albicans . A single pair of primers was designed in order to detect six of these SAP genes and, subsequently, to increase the sensitivity of the test . Detection of the PCR product by enzyme-linked immunosorbent assay was found to be as sensitive as Southern blotting with an SAP-labeled probe . The sensitivity of the assay was 1 cell/ml from serially diluted Candida cultures and 1 to 4 cells/ml from seeded blood specimens . The sensitivity and specificity of the present assay were tested in a retrospective study performed blindly with 156 clinical samples and were 100 and 98%, respectively, compared with the results of culture . For the subset of blood culture samples (n = 124), the sensitivity and the specificity were 100% . The two false-positive PCR samples came from patients treated with azole antifungal agents, indicating that PCR was probably able to detect damaged organisms that could not be recovered by culture.

J Clin Microbiol, 1998 Feb, 36(2), 367 - 74
Rapid differentiation of closely related Candida species and strains by pyrolysis-mass spectrometry and Fourier transform-infrared spectroscopy; Timmins EM et al.; Two rapid spectroscopic approaches for whole-organism fingerprinting of pyrolysis-mass spectrometry (PyMS) and Fourier transform-infrared spectroscopy (FT-IR) were used to analyze a group of 29 clinical and reference Candida isolates . These strains had been identified by conventional means as belonging to one of the three species Candida albicans, C . dubliniensis (previously reported as atypical C . albicans), and C . stellatoidea (which is also closely related to C . albicans) . To observe the relationships of the 29 isolates as judged by PyMS and FT-IR, the spectral data were clustered by discriminant analysis . On visual inspection of the cluster analyses from both methods, three distinct clusters, which were discrete for each of the Candida species, could be seen . Moreover, these phenetic classifications were found to be very similar to those obtained by genotypic studies which examined the HinfI restriction enzyme digestion patterns of genomic DNA and by use of the 27A C . albicans-specific probe . Both spectroscopic techniques are rapid (typically, 2 min for PyMS and 10 s for FT-IR) and were shown to be capable of successfully discriminating between closely related isolates of C . albicans, C . dubliniensis, and C . stellatoidea . We believe that these whole-organism fingerprinting methods could provide opportunities for automation in clinical microbial laboratories, improving turnaround times and the use of resources.

J Oral Pathol Med, 1998 Jan, 27(1), 4 - 7
Prevalence of Candida species in AIDS patients and HIV-free subjects in Thailand; Teanpaisan R et al.; The purpose of this study was to examine the prevalence of Candida species among groups of HIV-infected and HIV-free subjects in Thailand and to ascertain whether particular Candida species were associated with HIV infection . Oral rinse specimens were collected from 45 AIDS patients (CDC stage IV), 74 HIV-free healthy subjects, and 42 HIV-free patients who had clinical candidiasis . Yeasts recovered in culture were identified and quantified . The mean ages of the cohorts were 30.75+/-8.19 years (AIDS group), 28.50+/-7.98 (HIV-free healthy group) and 41.83+/-12.25 years (HIV-free candidiasis group) . Yeasts were isolated from 30/45 (66.66%, range 6.6x10(2)-5.7x10(6) CFU/ml) of the AIDS group, 8/74 (10.81%, range 8.0x10(1)-3.5x10(4) CFU/ml) of the HIV-free healthy group, and 24/42 (57.14%, range 1.0x10(2)-1.1x10(5) CFU/ml) of the HIV-free candidiasis group . There were statistically significant differences in the Candida colony counts between the AIDS group without oral candidiasis and the healthy group (P=0.0078) and between the AIDS group with candidiasis and the HIV-free, oral candidiasis group (P=0.0003) . Candida albicans was the most common species recovered from AIDS patients (29 out of 30; 96.66%)PIP: The association between Candida species and HIV serostatus in Thailand was compared in 45 patients hospitalized with AIDS-related conditions, 74 HIV-negative blood donors, and 42 HIV-negative individuals with clinical signs of oral candidiasis . Pseudomembranous and erythematous lesions were the most common clinical findings . After collection of oral rinse specimens by a single clinician, yeasts recovered in culture were identified and quantified . Yeasts were isolated from 30 AIDS patients (66.6%), 8 HIV-negative blood donors (10.81%), and 24 individuals in the HIV-free candidiasis group (57.14%) . Both the number of subjects who yielded Candida species and the mean number of Candida colonies were significantly higher in the AIDS group than in the HIV-free candidiasis group . C . albicans was the isolated yeast in 96.66% of AIDS patients, 79.16% of HIV-free candidiasis patients, and 100% of those in the healthy carrier group . Although C . albicans is a common and harmless commensal of the mucous membranes, it can cause severe mucosal or invasive disease in immunodeficient patients . It has been postulated that Candida strains that are nonpathogenic in healthy persons become pathogenic in HIV-infected individuals due to impaired host defense mechanisms .

J Infect Dis, 1998 Feb, 177(2), 347 - 54
Human immunodeficiency virus type 1 envelope protein gp120 impairs intracellular antifungal mechanisms in human monocytes; Pietrella D et al.; The key to success of fungal opportunistic pathogens in the immunocompromised host is related to survival inside phagocytic cells, which represent the first line of defense against microorganisms . The contribution of human immunodeficiency virus-1 recombinant envelope protein gp120 on effector functions of peripheral blood monocytes (PBM) against Candida albicans was investigated . gp120 binds CD4 receptors on PBM while not affecting the access of the fungus into the lysosome compartment . However, gp120 reduces the antifungal capacity of PBM . This phenomenon correlates with impaired oxygen-dependent antimicrobial machinery and reduced ability of phagolysosome acidification . The maintenance of phagolysosomal pH at approximately 6.2 restricts antimicrobial properties of the enzyme that work at a low pH, as evidenced by reduced antifungal capability of lysosomal protein extracted from gp120-treated PBM . These findings highlight gp120 perturbation of intracellular antimicrobial mechanisms of phagocytic cells and suggest a new aspect for gp120 in impairing immune functions.

J Antimicrob Chemother, 1997 Dec, 40(6), 889 - 93
Effects of in-vitro activity of miconazole and ketoconazole in phospholipid formulations; De Logu A et al.; Antifungal agents are often used in liposomal formulations in order to improve their pharmacological activity, but how vesicle inclusion can actually affect this is still not fully understood . We report here the results obtained from evaluation of the in-vitro activity against Candida albicans ATCC E10231 of miconazole and ketoconazole in various vesicular and non-vesicular preparations, obtained from egg and soya phospholipids, using time-kill curves . In most cases inclusion of miconazole or ketoconazole in liposomes led to a delayed and decreased activity of the drugs, with detectable differences among the various phospholipid concentrations and different liposomal preparations (small unilamellar vesicle, liposomes, multilamellar aggregates and broken liposomal structures) . The results obtained may be helpful in the study of new preparations of antifungal agents entrapped in liposomal structures.

J Antimicrob Chemother, 1997 Dec, 40(6), 855 - 62
Fluconazole versus nystatin in the prevention of candida infections in children and adolescents undergoing remission induction or consolidation chemotherapy for cancer; Groll AH et al.; An open, prospective, randomized pilot study was performed to assess the efficacy and safety of oral fluconazole 3 mg/kg once daily compared with oral nystatin 50,000 units/kg/day in four divided doses in preventing candida infections in 50 children undergoing remission induction or consolidation therapy for cancer . In 21 of 25 fluconazole-treated and 20 of 25 nystatin-treated patients the overall outcome of prophylaxis was clearly successful . Mild and transient oropharyngeal candidosis was observed in two and three patients in the fluconazole and nystatin groups respectively . One patient randomized to fluconazole and two patients randomized to nystatin required empirical treatment with amphotericin B and one patient assigned to fluconazole developed tissue-proven candida colitis . Initially non-colonized patients remained yeast-free throughout treatment with no differences between the two study arms . Initially colonized patients stayed colonized throughout treatment although at the end of the study, more patients randomized to nystatin were still harbouring yeasts (P = 0.05) . Almost exclusively, Candida albicans (95%) was isolated . A change in species was observed in one patient in each arm of the study . Candida krusei or Candida glabrata were not encountered . Transient elevations of hepatic transaminases were more common in the fluconazole group, although not statistically significant (28% vs 12%, P = 0.15) . Reversible grade I gastrointestinal and skin symptoms were observed in four patients randomized to fluconazole (16 vs 0%, P < 0.05) . Fluconazole was as safe and effective as nystatin in controlling yeast colonization and in preventing superficial and invasive candida infections and the empirical use of amphotericin B in children and adolescents undergoing intensive chemotherapy for cancer.

J Dent Assoc S Afr, 1995 Dec, 50(12), 601 - 4
Oral yeast flora of a Kalahari population; Blignaut E et al.; During an epidemiological survey of 181 individuals working or residing in the Kalahari National Gemsbok Park, swabs were taken from the dorsal surfaces of their tongues to determine a possible association between oral yeasts and clinically observed oral lesions as well as other underlying conditions detectable by serum chemistry . Identification of yeasts was performed with a commercially available identification system, namely the ATB 32C (Montalieu, Vercieu) . Yeasts were isolated from 30.4 per cent (n = 55) of individuals, of whom 43.6 per cent (n = 24) had only Candida albicans, 3.6 per cent (n = 2) had C . albicans together with other yeasts and 52.7 per cent (n = 29) had other yeasts . Many of these yeasts were not the commonly encountered clinical isolates . The results revealed a significant association (p < 0.02) between yeasts (n = 55) and low serum iron concentrations (n = 50) . A highly significant (p < 0.001) association was also found between smoking (n = 112) and the presence of clinically detectable oral lesions, notably leukoplakia (n = 21) and mucosal atrophy . The findings of this study reveal that a significant association exists between the oral yeast flora and serum iron and glucose, as well as between smoking and oral mucosal lesions.

J Infect, 1997 Nov, 35(3), 295 - 7
Hazards of inadequate fluconazole dosage to treat deep-seated or systemic Candida albicans infection; Flanagan PG et al.; We report three cases of deep-seated and systemic Candida albicans infection in which inadequate dosages of fluconazole were used, leading to breakthrough fungaemia, candidal osteomyelitis and endocarditis . The need to modify fluconazole dosage in patients receiving continuous venovenous haemofiltration is discussed.

Diagn Microbiol Infect Dis, 1997 Dec, 29(4), 227 - 31
Growth medium effect on the antifungal activity of LY 303366; Klepser ME et al.; The impact of growth medium selection on antifungal susceptibility testing has been well documented . Previously we described the antifungal characteristics of LY 303366 via time-kill curve methods using RPMI 1640 buffered with 0.165 M morpholinepropane-sulfonic acid as growth medium . The purpose of the current study was to compare the previously reported kill curve results with results obtained using antibiotic medium number three (AM #3) as growth medium . Antifungal activity was assessed via susceptibility testing and time-kill studies in both media . Two isolates each of Candida albicans, C . glabrata, and C . tropicalis were studied . MICs for the six isolates were found to be 10 to 100 times lower in AM #3 . Time-kill studies were conducted with multiples of the MIC ranging from 0.125 x MIC to 16 x MIC . LY 303366 exhibited fungicidal (> or = 3 log10 reduction in CFU) activity against all isolates in AM #3; however, fungicidal activity was noted only for three of the six isolates when tested in RPMI . Furthermore, the rate of fungicidal activity was more rapid when AM #3 was utilized . Not only were the rate and extent of activity influenced by choice of media, but the relationships between LY 303366 concentrations and activity were also found to be media dependent . The findings from this study serve to highlight further the importance of media selection for in vitro evaluation of antifungal activity . In vivo studies need to be conducted with LY 303366 to determine which media provides the best correlation between in vitro and in vivo findings.

Nippon Kyobu Geka Gakkai Zasshi, 1997 Nov, 45(11), 1862 - 5
{Candida costochondral osteomyelitis, report of a case and review of the literature}; Myojin K et al.; A 49-year-old man was admitted to the hospital with swelling and redness overlying the left anterior chest wall . He had been treated by percutaneous transluminal coronary recanalization for acute myocardial infarction with central venous catheter one year and four months earlier . Since then, he had had no symptoms . An incision and drainage was performed and specimen showed acute and chronic granulation tissue containing pus with involvement of underlying left third rib and cartilage . Candida albicans was cultured from the drainage specimen . Treatment with fluconazole was began . The lesion failed to clear following incision and drainage, continuing to exude pus, then open surgical excision and curettage of the cartilage and rib was performed . After 2 months of therapy, the lesion had resolved . This is a rare case of candida costochondral osteomyelitis without a definite proof of former hematogenous candida infection.

Proc Natl Acad Sci U S A, 1997 Dec 9, 94(25), 13816 - 9
Ascorbate recycling in human neutrophils: induction by bacteria; Wang Y et al.; Ascorbate (vitamin C) recycling occurs when extracellular ascorbate is oxidized, transported as dehydroascorbic acid, and reduced intracellularly to ascorbate . We investigated microorganism induction of ascorbate recycling in human neutrophils and in microorganisms themselves . Ascorbate recycling was determined by measuring intracellular ascorbate accumulation . Ascorbate recycling in neutrophils was induced by both Gram-positive and Gram-negative pathogenic bacteria, and the fungal pathogen Candida albicans . Induction of recycling resulted in as high as a 30-fold increase in intracellular ascorbate compared with neutrophils not exposed to microorganisms . Recycling occurred at physiologic concentrations of extracellular ascorbate within 20 min, occurred over a 100-fold range of effector/target ratios, and depended on oxidation of extracellular ascorbate to dehydroascorbic acid . Ascorbate recycling did not occur in bacteria nor in C . albicans . Ascorbate did not enter microorganisms, and dehydroascorbic acid entry was less than could be accounted for by diffusion . Because microorganism lysates reduced dehydroascorbic acid to ascorbate, ascorbate recycling was absent because of negligible entry of the substrate dehydroascorbic acid . Because ascorbate recycling occurs in human neutrophils but not in microorganisms, it may represent a eukaryotic defense mechanism against oxidants with possible clinical implications.

Clin Microbiol Rev, 1998 Jan, 11(1), 121 - 41
Serologic response to cell wall mannoproteins and proteins of Candida albicans; Martinez JP et al.; The cell wall of Candida albicans not only is the structure in which many biological functions essential for the fungal cells reside but also is a significant source of candidal antigens . The major cell wall components that elicit a response from the host immune system are proteins and glycoproteins, the latter being predominantly mannoproteins . Both the carbohydrate and protein moieties are able to trigger immune responses . Although cell-mediated immunity is often considered to be the most important line of defense against candidiasis, cell wall protein and glycoprotein components also elicit a potent humoral response from the host that may include some protective antibodies . Proteins and glycoproteins exposed at the most external layers of the wall structure are involved in several types of interactions of fungal cells with the exocellular environment . Thus, coating of fungal cells with host antibodies has the potential to influence profoundly the host-parasite interaction by affecting antibody-mediated functions such as opsonin-enhanced phagocytosis and blocking the binding activity of fungal adhesins for host ligands . In this review, the various members of the protein and glycoprotein fraction of the C . albicans cell wall that elicit an antibody response in vivo are examined . Although a number of proteins have been shown to stimulate an antibody response, for some of these species the response is not universal . On the other hand, some of the studies demonstrate that certain cell wall antigens and anti-cell wall antibodies may be the basis for developing specific and sensitive serologic tests for the diagnosis of candidasis, particularly the disseminated form . In addition, recent studies have focused on the potential for antibodies to cell wall protein determinants to protect the host against infection . Hence, a better understanding of the humoral response to cell wall antigens of C . albicans may provide the basis for the development of (i) effective procedures for the serodiagnosis of disseminated candidiasis and (ii) novel prophylactic (vaccination) and therapeutic strategies for the management of this type of infection.

Oral Dis, 1997 May, 3 Suppl 1, S102 - 9
Stable azole drug resistance associated with a substrain of Candida albicans from an HIV-infected patient; White TC et al.; Oral candidiasis is one of the earliest and most frequent complications of a failing immune system in HIV-infected individuals . For several years, oral candidiasis has been treated effectively with azole drugs, the one most frequently used is fluconazole . Unfortunately, extensive use of the drug for treatment and prophylaxis has led to treatment failure in an increasing number of patients . In most of these cases, strains of C . albicans isolated from the infection are less susceptible to fluconazole . The development of azole resistance in strains of C . albicans has been studied biochemically and more recently with molecular techniques . One excellent example of the development of azole resistance in C . albicans has been documented in a series of 17 C . albicans isolates from a single patient over a 2-year period . During this time, the patient experienced 14 episodes of oral candidiasis and was treated with increasing doses of fluconazole . Molecular and biochemical analyses confirms that the isolates are the same strain of C . albicans and that the resistance in these isolates is stable over 600 generations, suggesting that the changes in this strain are genetic in nature . In addition, the development of resistance is correlated with the identification of a substrain or variant of the original strain, as identified by restriction fragment length polymorphism (RFLP) analysis with the moderately repetitive probe, Ca3 . The analysis of this series of isolates demonstrates that azole drug resistance is associated with several small genetic changes, each of which contributes to the overall resistance of the strain . Clearly, continual use of azole drugs by a patient can select for genetic changes that render oral candidiasis refractory to treatment.

Oral Dis, 1997 May, 3 Suppl 1, S96 - 101
Molecular and phenotypic analysis of Candida dubliniensis: a recently identified species linked with oral candidosis in HIV-infected and AIDS patients; Coleman D et al.; The discovery and characterisation of a novel species of Candida, termed Candida dubliniensis, associated with oral candidosis in HIV-infected individuals is described . These organisms share several phenotypic characteristics in common with Candida albicans and Candida stellatoidea, including the ability to produce germ tubes and chlamydospores . However, in contrast to these latter two species, C . dubliniensis isolates produce abundant chlamydospores, which are often arranged in contiguous pairs, triplets and other multiples suspended from a single suspensor cell . They belong to C . albicans serotype A and exhibit atypical substrate assimilation profiles . Genomic DNA fingerprinting analysis with the C . albicans-specific probe 27A and five different oligonucleotide probes consisting of short repeat sequence-containing motifs, demonstrated that C . dubliniensis has a distinct genomic organisation relative to C . albicans and C . stellatoidea . This was confirmed by karyotype analysis and random amplified polymorphic DNA (RAPD) analysis . Comparison of 500 bp of the V3 variable region of the large ribosomal subunit genes from 14 separate C . dubliniensis isolates and the corresponding sequences from C . albicans, C . stellatoidea, C . tropicalis, C . glabrata, C . parapsilosis, C . kefyr and C . krusei demonstrated that the C . dubliniensis isolates formed a homogenous cluster (100% similarity), representing a discrete taxon within the genus Candida that was significantly different from the other species analysed.

Ann Thorac Surg, 1998 Jan, 65(1), 95 - 100
Secular trends in nosocomial bloodstream infections in a 55-bed cardiothoracic intensive care unit; Gordon SM et al.; BACKGROUND: Although bloodstream infections (BSIs) occur more frequently in intensive care unit patients than in ward patients, most studies of nosocomial BSIs in critically ill patients have not distinguished between intensive care unit populations beyond surgical, medical, and pediatric patients . METHODS: The primary objective of this study was to characterize the secular trends in rates of nosocomial BSIs for all pathogens among patients admitted to a busy cardiothoracic intensive care unit in a single tertiary care institution between January 1986 and December 1995 . Patients with nosocomial BSIs were identified through continual prospective surveillance . RESULTS: A total of 40,207 patients were admitted to the cardiothoracic intensive care unit during the 10-year study period, and 804 episodes of nosocomial BSIs among 681 patients were identified . The mean crude BSI infection rate was 6.0 per 1,000 patient-care days and increased linearly during the study period (range, 4.4 to 8.1 per 1000 patient-care days), and approached statistical significance (p value = 0.07) . The most common organisms causing BSIs were Staphylococcus aureus (12%), coagulase-negative staphylococci (11%), Candida albicans (11%), Pseudomonas aeruginosa (10%), and Enterococci (9%) . The leading sources of nosocomial BSIs were primary BSIs (33%), intravascular devices (27%), lower respiratory tract infections (17%), and surgical wound infections (12%) . The etiologic fraction or the proportion of deaths in cardiothoracic intensive care unit patients with BSIs was 15-fold higher than those patients without BSIs (37% versus 2.5%, p < 0.001) . CONCLUSIONS: Rates of nosocomial BSIs among patients in our cardiothoracic intensive care unit have increased linearly during the past decade and patients with nosocomial BSIs have an increased risk of in hospital mortality.

Rev Med Chir Soc Med Nat Iasi, 1996 Jan-Jun, 100(1-2), 167 - 71
{The reaction of 3-(R-phenyl)-6-hydrazine pyridazine with 5-methylisatin and 1-morpholinomethyl-5-methylisatin}; Dorneanu M et al.; This paper presents the synthesis of six hydrazones obtained by treating 5-methyl-isatin or 1-morpholino methyl-5-methyl-isatin with 3-(R-phenyl)-6-hydrazino-pyridazine (R = OCH3, Cl, Br) and two complex combination with copper, derived from 3-(p-anisyl-pyridazinyl)-hydrazone-5-methyl-indoline-2- one . The structure of the new compounds was confirmed by the results of the quantitative elementary and IR, UV-VIS spectral analysis . The biological tests point out that product VI, in which a copper atom binds two molecules of 3-(p-anisyl-pyridazinyl)-hydrazono-5-methyl-indoline-2-one, has a considerable antiinflammatory activity, giving a inflammation inhibition of 39, 6% . All the synthetized compounds have a moderate antimicrobial activity against Candida albicans.

Allerg Immunol (Paris), 1997 Oct, 29(8), 233 - 8
{Diagnostic contribution of abnormal delayed-type hypersensitivity to Candida albicans . Characterization test by activation of cells sensitized to successive dilutions of Candida}; Brunet JL et al.; By measuring the activation of different cell models (lymphocytes and lymphocytic subsets) in the presence of Candida albicans with flow cytometry reading, it is possible to show that successive dilutions of Candida albicans can lead to lymphocyte activation in abnormally-sensitized subjects . In a first trial, 10 subjects were tested in duplicate . The decrease of activity of the dilutions does not appear to be regular in relation to the progression of the dilutions . The activity of the dilutions wanes relatively rapidly with the first dilutions, then recurs later very distinctly, at the 6th dilution, then ebbs, then reappears in similar manner at the 9th, the 14th, and finally, the 19th dilution . Cell reactivity appears to differ depending on the subject . It can be represented through the calculated slope of the regression line, for each series of data . It therefore appears feasible to determine a threshold of reactivity and a scale of sensitivity, to make it possible to specify the degree of abnormal reactivity existing at a given time for a given subject . The constancy of the activity of the different dilutions tested, on 10 cultures of a single cell suspension, is especially well demonstrated in the second trial, showing unusually small standard deviations . Thus, the question arises as to the exact nature of the observed phenomenon and of its analysis from a physical-chemical point of view, with regard to the pharmacological effect of successive dilutions of Candida albicans.

Infect Immun, 1998 Feb, 66(2), 676 - 81
Proteolytic activation of the interleukin-1beta precursor by Candida albicans; Beausejour A et al.; Chronic inflammation rather than invasion is characteristic of some forms of superficial candidiasis such as denture stomatitis . We hypothesized that Candida albicans may play a critical role in the pathogenesis of inflammatory lesions observed in chronic candidiasis by activating the proinflammatory cytokine interleukin-1beta (IL-1beta) from epithelial stores of the precursor . The aim of this study was therefore to demonstrate the proteolytic cleavage and activation of the inactive precursor of IL-1beta (pro-IL-1beta) by C . albicans . After incubation of either blastospores or hyphae with the inactive precursor, proteolytic cleavage was monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis Western immunoblotting analysis, and the biological activity of the cleavage products was tested in a bioassay . We report here that late-stationary-growth-phase blastospores as well as hyphae of C . albicans, but not exponentially growing cells, can efficiently cleave pro-IL-1beta to yield fragments of molecular masses compatible with mature biologically active IL-1beta (17 to 19 kDa) . Assays conducted in the presence of selected proteinase inhibitors suggest that the cleavage of pro-IL-1beta involves the participation of one or more aspartyl proteinases . Cleavage products showed a dose-dependent IL-1beta-like activity in a thymocyte proliferation bioassay, which was inhibited by anti-IL-1beta neutralizing antibodies . The present data thus suggest a role for C . albicans proteinases in the activation and maintenance of the inflammatory response at epithelial surfaces.

FEMS Immunol Med Microbiol, 1997 Nov, 19(3), 223 - 30
Resistance to candidiasis and macrophage activity in chitin-treated mice; Rementeria A et al.; The effect of chitin, a polysaccharide of the cell wall of Candida albicans, on both the survival of C . albicans infected mice and the activity of the murine peritoneal macrophages has been studied . Pretreatment of mice with 30 mg kg(-1) C . albicans chitin enhanced the survival of the infected animals . The protective effect was concomitant with an enhancement of both phagocytic and candidacidal activities of the peritoneal macrophages . Chitin by itself did not induce the nitric oxide (NO) synthase in the macrophages, which remained at a level similar to that shown by the macrophages from untreated animals . The administration of 10 mg kg(-1) C . albicans chitin diminished the long term survival of the infected animals . This effect was coincident with a lower candidacidal activity and NO production by the macrophages of the chitin treated and infected animals, compared to the untreated infected animals.

FEMS Microbiol Lett, 1998 Jan 1, 158(1), 69 - 74
Characterisation of human steroid hormone transport mediated by Cdr1p, a multidrug transporter of Candida albicans, belonging to the ATP binding cassette super family; Krishnamurthy S et al.; Cdr1p, a multidrug transporter from a pathogenic yeast Candida albicans, confers resistance to several unrelated drugs including anti-Candida drugs . We demonstrate that Cdr1p can specifically transport human steroid hormones namely beta-estradiol and corticosterone . Saccharomyces cerevisiae transformant S-12, harbouring the CDR1 gene, accumulated about 3-fold less {3H} beta-estradiol and about 2-fold less {3H}corticosterone than the non-transformed strain . When CDR1 was expressed in AD strain (AD-CDR1) which had seven ATP binding cassette (ABC) superfamily of putative transporter genes disrupted, the net accumulation of these hormones as compared to S-12 was significantly lower . Efflux of beta-estradiol and corticosterone was inhibited by a 100-fold higher (200 nM) concentration of beta-estradiol, corticosterone, ergosterol or dexamethasone, but progesterone which could not be transported by Cdr1p did not affect the efflux and thus accumulation . Interestingly, some of the drugs viz . cycloheximide, chloramphenicol, fluconazole and o-phenanthroline, to which CDR1 confers resistance, could also prevent efflux and enhance accumulation to some extent . In conclusion, we show that human steroid hormones could be the substrates for Cdr1p and the energy dependent transport mediated by it is specific for estradiol and corticosterone.

Pharm Res, 1997 Dec, 14(12), 1765 - 71
The effects of hexetidine (Oraldene) on the adherence of Candida albicans to human buccal epithelial cells in vitro and ex vivo and on in vitro morphogenesis; Jones DS et al.; PURPOSE: This study reports the effects of hexetidine (Oraldene) on two virulence attributes of Candida albicans, namely, in vitro and ex vivo adherence of yeast cells to buccal epithelial cells (BEC) and in vitro morphogenesis . METHODS: The effects of hexetidine treatment of either yeast cells (stationary and exponential phases) or BEC on Candidal adherence, in terms of viable and non-viable adherent yeast cells, were evaluated using an acridine orange stain in conjunction with fluorescence microscopy . Ex vivo anti-adherence effects were determined by rinsing BEC in vivo with hexetidine (0.1%), removal of BEC after defined periods and inclusion in the adherence assay . The effects of hexetidine on morphogenesis were evaluated using light microscopy . Yeast cell viability following exposure to a range of concentration of hexetidine (0.005-0.1% v/v) for defined periods was determined following serial dilution and enumeration on solid media . RESULTS: Treatment of stationary and exponential phase yeast cells or BEC with hexetidine (0.1%) for a range of times (10-300 s) or, alternatively, with a range of concentrations of hexetidine (0.005-0.1%) for a fixed time (30s) significantly decreased the resultant Candidal/ epithelial adhesion . No correlations were observed between reduced adherence and either time of treatment or hexetidine concentration . In vivo treatment of BEC with hexetidine (0.1%) for 30s resulted in prolonged and significant reductions in the ex vivo adherence of both viable and non-viable yeast cells for periods of up to (and including) four hours post-rinsing . Treatment of C . albicans blastospores with hexetidine (0.05, 0.1% v/v) for 10s and 30s totally inhibited Candida morphogenesis, whereas treatment with lower antiseptic concentrations significantly reduced the extent of Candida morphogenesis and the rate of hyphal development . The effects of hexetidine on yeast cell viability were both concentration and time-dependent . CONCLUSIONS: The reduced adherence of C . albicans to BEC and the modification or inhibition of morphogenesis following exposure to hexetidine suggests a clinical role for hexetidine in the prophylaxis of both superficial candidosis and the systemic complications resulting from invasion of sub-epithelial tissue.

Clin Infect Dis, 1997 Dec, 25(6), 1359 - 62
Pacemaker endocarditis due to Candida albicans: case report and review; Joly V et al.; We describe a case of pacemaker endocarditis due to Candida albicans in a patient who responded favorably to combined surgical and antifungal therapy . Only five cases of candidal pacemaker endocarditis have been reported previously . We review these five cases and discuss the clinical presentation and therapy for this disease in comparison with candidal prosthetic valve endocarditis.

Crit Rev Biochem Mol Biol, 1997, 32(5), 405 - 35
Coregulation of starch degradation and dimorphism in the yeast Saccharomyces cerevisiae; Vivier MA et al.; Saccharomyces cerevisiae, the exemplar unicellular eukaryote, can only survive and proliferate in its natural habitats through constant adaptation within the constraints of a dynamic ecosystem . In every cell cycle of S . cerevisiae, there is a short period in the G1 phase of the cell cycle where "sensing" transpires; if a sufficient amount of fermentable sugars is available, the cells will initiate another round of vegetative cell division . When fermentable sugars become limiting, the yeast can execute the diauxic shift, where it reprograms its metabolism to utilize nonfermentable carbon sources . S . cerevisiae can also initiate the developmental program of pseudohyphal formation and invasive growth response, when essential nutrients become limiting . S . cerevisiae shares this growth form-switching ability with important pathogens such as the human pathogen, Candida albicans, and the corn smut pathogen Ustilago maydis . The pseudohyphal growth response of S . cerevisiae has mainly been implicated as a means for the yeast to search for nutrients . An important observation made was that starch-degrading S . cerevisiae strains have the added ability to form pseudohyphae and grow invasively into a starch-containing medium . More significantly, it was also shown that the STA1-3 genes encoding three glucoamylase isozymes responsible for starch hydrolysis in S . cerevisiae are coregulated with a gene, MUC1, essential for pseudohyphal and invasive growth . At least two putative transcriptional activators, Mss10p and Mss11p, are involved in this regulation . The Muc1p is a putative integral membrane-bound protein similar to mammalian mucin-like proteins that have been implicated in the ability of cancer cells to invade other tissues . This provided us with an excellent example of integrative control between nutrient sensing, signaling, and differential development.

Arzneimittelforschung, 1997 Dec, 47(12), 1393 - 7
Synthesis and microbiological activity of 5(or 6)-methyl-2-substituted benzoxazole and benzimidazole derivatives; Oren I et al.; The synthesis and microbiological activity of a new series of 5(or 6)-methyl-2-substituted benzoxazoles (IVa-n) and benzimidazoles (Va-h) were described . The in vitro microbiological activity of the compounds was determined against gram-positive, gram-negative bacteria and the yeast Candida albicans in comparison to reference drugs . Microbiological results indicated that the derivatives possessed a broad spectrum of activity against the tested microorganisms and the compounds IVa-g, IVi-k, IVn, Vb-c and Vh showed significant activity against Pseudomonas aeruginosa having MIC values of 25 micrograms/ml, providing higher potencies than the reference drugs tetracycline and streptomycin . Moreover, the derivative 5-methyl-2-(p-chlorobenzyl)benzoazole (IVb) possessed the same potency against Candida albicans as the reference drugs oxiconazole and haloprogin having a MIC value of 6.25 mg/ml . However, none of the derivatives showed a better spectrum of activity than the reference drugs.

Antimicrob Agents Chemother, 1998 Jan, 42(1), 147 - 50
KY-62, a polyene analog of amphotericin B, for treatment of murine candidiasis; Graybill JR et al.; KY-62 is a water-soluble analog of amphotericin B . In vitro testing of five clinical isolates of Candida albicans showed KY-62 to have potency similar to that of amphotericin B . KY-62 was administered to mice infected intravenously with C . albicans . In vivo, KY-62 was effective in immunocompetent mice, with potency similar to that of amphotericin B . KY-62 was well tolerated up to 30 mg/kg of body weight per dose, an amount that would be lethal with amphotericin B . KY-62 was less effective in mice rendered neutropenic with 5-fluorouracil . The addition of flucytosine had little effect . KY-62 may have potential for clinical development.

J Am Dent Assoc, 1998 Jan, 129(1), 78 - 83
Case report . Use of an argon laser to treat drug-induced gingival overgrowth; Mattson JS et al.; This article explores the use of an argon laser to treat severe drug-induced gingival overgrowth . The patient was being treated with phenytoin (Dilantin, Parke-Davis), cyclosporine and a calcium channel blocker . He had undergone a kidney transplantation and had insulin-dependent diabetes mellitus . He had severe gingival overgrowth, which prevented him from wearing his removable prostheses, and a superimposed Candida albicans infection . An argon laser was used to excise the gingival overgrowth so new maxillary and mandibular prostheses could be fabricated.

Eur J Clin Microbiol Infect Dis, 1997 Nov, 16(11), 848 - 51
Increased adherence of Candida albicans to buccal epithelial cells from patients with AIDS; Schwab U et al.; The adherence of six clinical Candida albicans isolates to buccal epithelial cells obtained from AIDS patients, solid organ transplant recipients and healthy individuals was compared . It was shown that Candida albicans bound in significantly greater numbers to epithelial cells obtained from AIDS patients than to those from healthy individuals or transplant patients, and that the adherence capacity varied among the strains tested.

J Bacteriol, 1998 Jan, 180(2), 282 - 9
Cloning and characterization of PRA1, a gene encoding a novel pH-regulated antigen of Candida albicans; Sentandreu M et al.; Candida albicans is an opportunistic fungal pathogen of humans . The cell wall of the organism defines the interface between the pathogen and host tissues and is likely to play an essential and pivotal role in the host-pathogen interaction . The components of the cell wall critical to this interaction are undefined . Immunoscreening of a lambda expression library with sera raised against mycelial cell walls of C . albicans was used to identify genes encoding cell surface proteins . One of the positive clones represented a candidal gene that was differentially expressed in response to changes in the pH of the culture medium . Maximal expression occurred at neutral pH, with no expression detected below pH 6.0 . On the basis of the expression pattern, the corresponding gene was designated PRA1, for pH-regulated antigen . The protein predicted from the nucleotide sequence was 299 amino acids long with motifs characteristic of secreted glycoproteins . The predicted surface localization and N glycosylation of the protein were directly demonstrated by cell fractionation and immunoblot analysis . Deletion of the gene imparted a temperature-dependent defect in hypha formation, indicating a role in morphogenesis . The PRA1 protein was homologous to surface antigens of Aspergillus spp . which react with serum from aspergillosis patients, suggesting that the PRA1 protein may have a role in the host-parasite interaction during candidal infection.

FEMS Microbiol Lett, 1997 Dec 15, 157(2), 273 - 8
Cloning of a cDNA fragment encoding part of the protein moiety of the 58-kDa fibrinogen-binding mannoprotein of Candida albicans; Lopez-Ribot JL et al.; Immunoscreening of a Candida albicans expression library with antibodies against the 58 kDa fibrinogen-binding mannoprotein (mp58) of the fungus resulted in the isolation of clones encoding the protein moiety of this molecule . Sequence of the 0.9 kb cDNA of one of the clones selected for further analysis, revealed an open reading frame coding for 292 amino acids, which displays sequence similarity to proteins belonging to a family of immunodominant antigens of Aspergillus spp . The gene corresponding to this cDNA was named FBP1 (fibrinogen-binding protein) . These results represent the first report on the identification of C . albicans genes encoding surface receptors for host proteins.

Mycopathologia, 1997, 138(2), 57 - 64
Influences of immunosuppressive agents, FK506 and cyclosporin on systemic Candida albicans infection in mice; Ito E et al.; The effects of the immunosuppressive agents FK506 (tacrolimus) and cyclosporin (CyA) on Candida albicans infection in mice were compared with those of cyclophosphamide . FK506 and CyA did not exacerbate C . albicans infection in mice when the effects were determined on the basis of survival ratio and colony forming units (CFU) in the kidney, although cyclophosphamide (CY) impaired the host defence mechanisms of mice against C . albicans infection . The effects of FK506 and CyA on the body weight of mice, histopathological changes of lymphoid tissues and formation of granulomas in kidney were also studied in comparison with those of CY.

Arzneimittelforschung, 1997 Nov, 47(11), 1263 - 5
Correlation between antifungal activity and hydrophobicity of imidazole antifungal agents; Hashiguchi T et al.; To examine the effects of the affinity of antifungal imidazole agents to cell membranes on the in vitro antifungal activity, the correlation between antifungal activity and hydrophobicity of these drugs was investigated . The capacity factor (k') in HPLC was used as a parameter of hydrophobicity, and the minimum inhibitory concentration (MIC) as a parameter of antifungal activity . Consequently, a significant portion correlation was noted in the case of Candida albicans . However, no obvious correlation was noted for Trichophyton mentagrophytes and Trichophyton rubrum . Therefore, the mechanism of action of imidazole derivatives for Candida albicans was considered to mainly consist in the membrane action.

Rev Soc Bras Med Trop, 1997 Nov-Dec, 30(6), 441 - 6
Flow cytometric quantitation of phagocytosis in heparinized complete blood with latex particles and Candida albicans; Egido JM et al.; We report a rapid method for the flow cytometric quantitation of phagocytosis in heparinized complete peripheral blood (HCPB), using commercially available phycoerythrin-conjugated latex particles of 1 micron diameter . The method is faster and shows greater reproducibility than Bjerknes' (1984) standard technique using propidium iodide-stained Candida albicans, conventionally applied to the leukocytic layer of peripheral blood but here modified for HCPB . We also report a modification of Bjerknes' Intracellular Killing Test to allow its application to HCPB.

Infect Immun, 1998 Jan, 66(1), 191 - 6
Endothelial cell injury caused by Candida albicans is dependent on iron; Fratti RA et al.; Although it is known that Candida albicans causes endothelial cell injury, in vitro and in vivo, the mechanism by which this process occurs remains unknown . Iron is critical for the induction of injury in many types of host cells . Therefore, we investigated the role of iron in Candida-induced endothelial cell injury . We found that pretreatment of endothelial cells with the iron chelators phenanthroline and deferoxamine protected them from candidal injury, even though the organisms germinated and grew normally . Loading endothelial cells with iron reversed the cytoprotective effects of iron chelation . Moreover, chelation of endothelial cell iron significantly reduced phagocytosis of C . albicans by these cells, while candidal adherence to chelator-treated endothelial cells was slightly enhanced . Since endothelial cell phagocytosis of C . albicans is required for endothelial cell injury to occur, inhibition of phagocytosis is likely the principal mechanism of the cytoprotective effects of iron chelation . The production of toxic reactive oxygen intermediates by host cells is known to be inhibited by iron chelation . Therefore, we investigated whether treating endothelial cells with antioxidants could mimic the cytoprotective effects of iron chelation . Neither extracellular nor membrane-permeative antioxidants reduced candidal injury of endothelial cells . Furthermore, depleting endothelial cells of the endogenous antioxidant glutathione did not render them more susceptible to damage by C . albicans . These results suggest that candidal injury of endothelial cells is independent of the production of reactive oxygen intermediates and that the cytoprotective effects of iron chelation are not due to inhibition of the synthesis of these toxic intermediates.

Wiad Lek, 1997, 50 Suppl 1 Pt 2, 259 - 63
{Changes in tissue plasminogen activator in experimental peritonitis caused by pathogenic factors}; Karon J et al.; Significantly higher tissue plasminogen activator activities were found in the abdominal exudate of rats with peritonitis caused by Escherichia coli than in rats with peritonitis caused by Staphylococcus aureus and Candida albicans . The changes of intraabdominal fibrinolysis may be depended on the type of microorganism.

Infect Immun, 1998 Jan, 66(1), 145 - 50
Interleukin-15 augments superoxide production and microbicidal activity of human monocytes against Candida albicans; Vazquez N et al.; Interleukin-15 (IL-15) is a newly described cytokine that shares biological activities with IL-2 . We report here results demonstrating the ability of IL-15 to enhance superoxide production and antifungal activity of human monocytes . After 18 and 48 h of treatment with IL-15, human elutriated monocytes manifested enhanced superoxide production in response to either phorbol myristate acetate or opsonized Candida albicans blastoconidia . Similar results were obtained when monocytes were treated with IL-2, but to a lesser extent . Combination studies with IL-15 and IL-2 showed no additive or synergistic effects . Following incubation of monocytes with IL-15 for 18 h, there was no significant increase in mRNA transcripts for components of the NADPH oxidase complex, p40-phox, p47-phox, and gp91-phox, suggesting a posttranscriptional modulation of enhanced superoxide production . Antibodies against the gamma chain of the IL-2 receptor and, to a lesser extent, against the beta chain partially abrogated the IL-15-mediated enhanced superoxide production . Additionally, human monocytes showed enhanced killing activity against C . albicans after 18 h of incubation with IL-15 or IL-2, but this treatment did not enhance the ability of these cells to phagocytose the organism . In addition, the enhanced fungicidal activity seen after 18 h of treatment was no longer detectable after 48 h of cytokine treatment . Culture supernatants from the IL-15-treated monocytes were assayed for the presence of other proinflammatory cytokines . IL-15 treatment did not induce the release of detectable levels of tumor necrosis factor alpha, IL-1beta, or IL-12 . Our results indicate that IL-15 upregulates the microbicidal activity of human monocytes against C . albicans.

Infect Immun, 1998 Jan, 66(1), 140 - 4
Binding of Candida albicans to immobilized amino acids and bovine serum albumin; Hawser SP et al.; In this study, we examined the binding of Candida albicans synchronized yeast-phase cells to plastic, immobilized amino acids and bovine serum albumin (BSA) and quantified the binding by using an XTT tetrazolium salt assay and absorbance determination . Our results show that C . albicans binds efficiently and specifically to several nonpolar aliphatic amino acids and positively charged amino acids and to BSA immobilized on tissue culture plastic but not to polar uncharged, negatively charged, or aromatic amino acids . Adhesion of yeasts to immobilized amino acids was not affected by preincubation of cells with BSA, whereas binding to immobilized BSA was affected by preincubation of yeasts with alanine, proline, and leucine but not by arginine or lysine . The ability to distinguish the chirality of these amino acids was also examined by using both the D and L amino acid configurations, and the results show that C . albicans yeasts recognize only the L configuration of these amino acids . The observations that C . albicans specifically binds to certain amino acids indicate that these amino acids may prove useful tools for studying the binding interactions of C . albicans yeasts with host proteins such as components of the extracellular matrix.

J Bacteriol, 1998 Jan, 180(1), 163 - 6
Defects in assembly of the extracellular matrix are responsible for altered morphogenesis of a Candida albicans phr1 mutant; Popolo L et al.; Analysis of Candida albicans cells using antibodies directed against Gas1p/Ggp1p, Saccharomyces cerevisiae homolog of Phr1p, revealed that Phr1p is a glycoprotein of about 88 kDa whose accumulation increases with the rise of external pH . This polypeptide is present both in the yeast form and during germ tube induction . In the Phr1- cells at pH 8 the solubility of glucans in alkali is greatly affected . In the parental strain the alkali-soluble/-insoluble glucan ratio shows a 50% decrease at pH 8 with respect to pH 4.5, whereas in the null mutant it is unchanged, indicating the lack of a polymer cross-linker activity induced by the rise of pH . The mutant has a sixfold increase in chitin level and is hypersensitive to calcofluor . Consistently with a role of chitin in strengthening the cell wall, Phr1- cells are more sensitive to nikkomycin Z than the parental strain.

Microbiology, 1997 Dec, 143 ( Pt 12), 3757 - 65
Influence of Na+ and anions on the dimorphic transition of Candida albicans; Northrop FD et al.; The effect of Na+ (Cl- or gluconate salt) on growth and dimorphic potential of the pathogenic yeast Candida albicans has been examined . Profiles of germ tube formation as a function of salt addition, pH and temperature indicated Na+ inhibition of germ tube outgrowth at high ambient pH (pH 8.0) which was exacerbated by replacement of Cl- with gluconate (as an impermeant analogue) . At acidic pH (pH 5.5) and permissive temperature (37 degrees C), gluconate alone promoted the dimorphic transition . Rates of glucose-induced medium acidification and plasma membrane H(+)-ATPase activity have been measured to assess whether salt treatments could retard the cytoplasmic alkalinization known to precede germ tube formation . The precise site of Na+ action remains unclear but the anion effects may be interpreted in terms of anion-exchanger and channel activity acting to modulate cytosolic pH.

Microbiology, 1997 Dec, 143 ( Pt 12), 3747 - 56
A neutral trehalase gene from Candida albicans: molecular cloning, characterization and disruption; Eck R et al.; A neutral trehalase gene, NTC1, from the human pathogenic yeast Candida albicans was isolated and characterized . An ORF of 2724 bp was identified encoding a predicted protein of 907 amino acids and a molecular mass of 104 kDa . A single transcript of approximately 3.2 kb was detected by Northern blot analysis . Comparison of the deduced amino acid sequence of the C . albicans NTC1 gene product with that of the Saccharomyces cerevisiae NTH1 gene product revealed 57% identity . The NTC1 gene was localized on chromosome 1 or R . A null mutant (delta ntc1/delta ntc1) was constructed by sequential gene disruption . Extracts from mutants homozygous for neutral trehalase deletion had only marginal neutral trehalase activity . Extracts from heterozygous mutants showed intermediate activities between extracts from the wild-type strain and from the homozygous mutants . The null mutant showed no significant differences in pathogenicity as compared to the wild-type strain in a mouse model of systemic candidiasis . This result indicates that the neutral trehalase of C . albicans is not a potential target for antifungal drugs.

Semin Cutan Med Surg, 1997 Dec, 16(4), 265 - 72
Oral cavity complications of bone marrow transplantation; Eisen D et al.; Bone marrow transplantation, once regarded as experimental, has evolved into a standard treatment for a variety of malignancies . Considerable advances have been made in histocompatibility typing, pretransplantation chemotherapy, and posttransplantation immunosuppressive therapy as well as prophylaxis and treatment of infections . Oral complications develop in almost all patients, and their early recognition may result in the institution of prompt treatment and prolonged survival . Mucositis, often severe and extremely painful, develops in more than three quarters of bone marrow transplant recipients, and its prevention, unfortunately, remains unsatisfactory . Herpes simplex virus and Candida albicans account for most oral infections, although their incidence has been dramatically reduced by the institution of prophylactic agents . Graft versus host disease continues to be a significant complication of marrow transplantation, and the detection of commonly occurring oral changes may support its diagnosis.

Proc Soc Exp Biol Med, 1998 Jan, 217(1), 81 - 8
Exposure of macrophages to an enzymatically inactive macrophage mannose receptor ligand augments killing of Candida albicans; Gelderman MP et al.; Macrophages (Mphi) are involved in host defenses against opportunistic pathogens . Previous studies by the present investigators indicate that Mphi exposed to enzymatically active myeloperoxidase (MPO), exhibited both increased phagocytosis and killing of Candida albicans . The purpose of this study was to determine if enzymatically inactive Mphi-mannose receptor (MMR) ligands could function similarly . Resident murine peritoneal Mphi were exposed to the MMR ligands, mannosylated bovine serum albumin (mBSA), and enzymatically inactive myeloperoxidase (iMPO), followed by exposure to opsonized C . albicans . Both mBSA and iMPO induced a slight increase in the number of phagocytizing cells; however, candidacidal activity was significantly higher in treated cultures compared to controls (P < or = 0.001) . The production of reactive oxygen intermediates (ROI) was detected using chemiluminescence . After employment of ROI scavengers, a decrease in candidacidal activity was observed . The data suggest that MMR-ligand interaction alone is sufficient to significantly enhance the candidacidal activity of Mphi via ROI, and that iMPO which is released at a site of inflammation induces Mphi-mediated killing of microorganisms . These findings indicate a previously unrecognized role of iMPO.

J Trauma, 1997 Dec, 43(6), 894 - 8
The effect of blood transfusion on susceptibility to bacterial infection in genetically defined mouse models; Eaves-Pyles T et al.; BACKGROUND: Blood transfusions suppress immune function and increase susceptibility to infection, but the effects are not consistent . STUDY DESIGN AND METHODS: Genetically defined mouse strains with the same or different haplotypes were used as blood transfusion recipients and donors . Transfused animals were subjected to cecal ligation and puncture (CLP) and followed for survival or were injected intravenously with Candida albicans to follow clearance of the Candida from the kidneys . RESULTS: BALB/c (H-2d) mice transfused with C3H/HeJ (H-2k) or DBA/2 (H-2d) blood followed by CLP showed significantly lower survival (7 and 10%) than mice transfused with syngeneic blood (61%) or saline controls (56%) . Lower survival was also observed in C3H/HeJ (H-2k) mice transfused with BALB/c (H-2d) blood and subjected to CLP (25%) compared with syngeneic transfusion (80%) or saline controls (70%) . C57BL/6J (H-2b) mice showed minimal increases in mortality after CLP after transfusion with blood from C3H/HeJ (H-2k) (60% survival), DBA/2 (H-2d) (70% survival), or BALB/c (H-2d) mice (90% survival) . When C . albicans was infused intravenously into transfused mice, a similar pattern of altered resistance to infection was found . CONCLUSION: The ability of blood transfusions to increase susceptibility to bacterial infection appears to be dependent on genetic factors unrelated to the major haplotype.

J Infect Dis, 1998 Jan, 177(1), 175 - 81
Recombinant murine granulocyte colony-stimulating factor protects against acute disseminated Candida albicans infection in nonneutropenic mice; Kullberg BJ et al.; The effect of recombinant granulocyte colony-stimulating factor (rG-CSF) on acute disseminated Candida albicans infection in nonneutropenic mice was investigated . Mice treated with a single dose of rG-CSF showed a significantly reduced mortality (28% vs . 90%; P < .001) . The outgrowth of C . albicans from the kidneys, spleens, and livers of rG-CSF-treated mice was significantly reduced (log cfu/g of kidney, 5.54 vs . 7.13; P < .001), as were circulating tumor necrosis factor-alpha and interleukin-1beta . After rG-CSF, the kidneys showed fewer infectious infiltrates, enhanced granulocyte influx, and almost complete absence of hyphal outgrowth . During peritoneal C . albicans infection, rG-CSF enhanced influx of granulocytes to the site of infection, and exudate granulocytes showed increased oxygen radical production . These results indicate that rG-CSF enhances host resistance to disseminated candidiasis in nonneutropenic mice through activation of granulocytes and their recruitment to the site of infection.

Mycoses, 1997, 40 Suppl 1, 53 - 5
{Use of fluconazole as antimycotic prophylaxis in radiotherapy of patients with head and neck tumors}; Mucke R et al.; The aim of the present study was to investigate the incidence of Candida stomatitis and resulting interruptions in radiation therapy in 50 patients suffering from squamous cell carcinomas of head and neck region receiving fluconazole (100 mg/d) in comparison to a historical control group without specific prophylaxis . 20 of the control patients (40%) demonstrated Candida stomatitis with 7 of them (14%) requiring interruptions of radiation therapy . In contrast, none of the patients with fluconazole had evidence of Candida stomatitis and subsequent interruption of anticancer therapy . Laboratory monitoring for the presence of Candida species was performed in 15 other patients before and after therapy with fluconazole . Candida albicans was identified less frequently after therapy when compared to the pretreatment status . However, C . glabrata and C . krusei were isolated in some of the patients probably due to decreased drug susceptibility of these species . The results demonstrate the clinical usefulness of prophylactic fluconazole applications in patients suffering from head and neck tumors with the aim to reduce Candida stomatitis and resulting interruptions in radiation therapy.

Mycoses, 1997, 40 Suppl 1, 47 - 52
{Evaluation of a breakpoint test for determination of fluconazole susceptibility of yeasts}; Fegeler W; Using the breakpoint test at 1 g/ml and 4 g/ml fluconazole, a minimal inhibitory concentration (MIC) of < or = 4 g/ml fluconazole was determined against 78.5% of the 1254 clinical yeast isolates . When compared with the micro broth dilution test, none of a subset of 128/1254 strains had a higher MIC in the dilution test than in the breakpoint test, however, in 43.0% of the 128 strains the MIC was lower in the micro broth dilution test when compared to the MIC of the breakpoint test . In a subset of 94 strains with an MIC of > 4 g/ml fluconazole determined in the breakpoint test, the elevated MIC could be confirmed only in 45.7% of the strains when using the micro broth dilution test . The percentage of breakpoint test confirmation as well as the number of strains with decreased susceptibility towards fluconazole (> 4 g/ml) were species dependent, thus, the number of decreased-susceptible Candida albicans strains was smaller than that of C . glabrata or other Candida species such as C . krusei, C . inconspicua and some C . tropicalis strains . The breakpoint test allows to identify susceptible strains with a high accuracy . Strains with an MIC > 4 g/ml fluconazole should be tested in the micro dilution test to confirm decreased susceptibility and thus to indicate the need for higher dosage of fluconazole or a change of the antifungal therapy . The breakpoint test proved to be a rapid and reliable screening test.

Dermatology, 1997, 195(3), 235 - 8
Perioral dermatitis in children--clinical presentation, pathogenesis-related factors and response to topical metronidazole; Boeck K et al.; BACKGROUND: Perioral dermatitis, a common skin disorder in young women, is rarely described in children . OBJECTIVE: This study elaborates the clinical features of perioral dermatitis in children as well as possible pathogenetic mechanisms and the response to topical metronidazole . METHODS: Seven children (4 females, 3 males between 4 and 12 years of age) were evaluated and dermatological examination was carried out . Pretreatment with topical corticosteroids was documented . Skin prick test with a panel of six common aeroallergens was performed in all children . All children were screened for gastrointestinal colonization with Candida albicans . Patients were treated with topical metronidazole 1% during the first 2 weeks . From the 3rd week on 2% metronidazole was used . RESULTS: In all but one child topical corticosteroids had been used in the face prior to the first presentation at our outpatient department suggesting a possible pathogenetic role . An association with atopy or intestinal candida colonization was not found . In all children skin lesions resolved after 3-6 months . The children remained free of symptoms over an observation period of 2 years . CONCLUSION: Perioral dermatitis has to be considered as differential diagnosis in children presenting with erythematous papules and papulovesicles in typical locations . Metronidazole proved to be effective and safe in the treatment of perioral dermatitis in children . Atopy and gastrointestinal colonization with C . albicans do not seem to play a role in the pathogenesis of perioral dermatitis.

Dermatology, 1997, 195(3), 220 - 3
Skin surface lipids inhibit adherence of Candida albicans to stratum corneum; Law S et al.; BACKGROUND: Candida albicans (CA) infections represent a significant threat to the health of immunocompromised individuals . The initial step in the establishment of a CA infection is adherence of the organism to an epithelial surface . METHODS: An in vitro model for studies on adherence of CA to keratinized surfaces has been developed and used to test the hypothesis that specific lipids can modulate adherence of this organism . Porcine stratum corneum (SC) disks were incubated with candidal suspensions, after which unattached cells were washed away . Adherent cells were stained and counted using light microscopy . RESULTS: Attachment of three pathogenic CA isolates was significantly greater than attachment of commensal strains of either CA or Candida parapsilosis . Removal of lipid from the SC lead to a doubling of the number of adherent organisms, whereas additional skin lipid inhibited adherence . Squalene, wax esters, cholesterol esters and triglycerides had no effect on adherence, but fatty acids, sterols and ceramides inhibited adherence significantly . CONCLUSIONS: Specific epithelial lipids can modulate adherence of CA to keratinized epithelial surfaces.

AIDS, 1997 Dec, 11(15), 1839 - 44
Clinically significant azole cross-resistance in Candida isolates from HIV-positive patients with oral candidosis; Cartledge JD et al.; OBJECTIVES: To determine the proportion of fluconazole-resistant Candida albicans isolates that have clinically significant cross-resistance to itraconazole or ketoconazole, that is sufficient to result in failure of these agents at their standard doses (200 and 400 mg daily for 7 days, respectively) . METHODS: Seven hundred C . albicans isolates from HIV-positive patients with oral candidosis underwent susceptibility testing using a relative growth method, for which cut-off values corresponding to clinical drug failure have been established . RESULTS: A total of 431 isolates were fully azole-susceptible and three main resistance patterns were detected: isolates resistant to fluconazole alone (n = 100); isolates resistant to fluconazole and ketoconazole but susceptible to itraconazole (n = 94); and isolates resistant to all three drugs (n = 50) . No isolates were consistently resistant to ketoconazole without being fluconazole-resistant, and no itraconazole resistance was detected without ketoconazole resistance . Resistance to fluconazole alone was more common in specimens obtained soon after first clinical fluconazole failure, whereas specimens from patients with a longer history of fluconazole-unresponsive candidosis were more likely to be infected with cross-resistant isolates . Median days of prior azole exposure and cumulative fluconazole dose were significantly less for those with isolates resistant to fluconazole alone than for those with ketoconazole cross-resistant isolates, who had received less azole therapy and smaller cumulative fluconazole doses than those with isolates cross-resistant to all three drugs (although not statistically significant) . After the diagnosis of fluconazole-unresponsive candidosis, increasing cumulative doses of itraconazole solution were associated with increasing likelihood of cross-resistance . CONCLUSIONS: Clinically significant cross-resistance to other azoles may occur in fluconazole-resistant isolates of C . albicans, although initially most isolates are not cross-resistant and the detection of cross-resistant isolates is associated with a history of greater prior azole exposure . Patients who have been treated for fluconazole-resistant candidosis for longer and with greater cumulative doses of itraconazole solution tend to become infected with increasingly cross-resistant isolates of C . albicans.

Rev Clin Esp, 1997 Jul, 197(7), 500 - 1
{Spontaneous peritonitis caused by ascitic fluid with Candida albicans}; de Luis D et al.; Two cases are reported of spontaneous ascitic fluid Candida albicans peritonitis in two patients with liver cirrhosis secondary to HBV . In non of the two cases was there evidence of findings associated with secondary peritonitis, where ascitic fluid Candida infection has usually been reported.

Biotech Histochem, 1997 Sep, 72(5), 268 - 72
The use of dimethylsulfoxide for fixation of yeasts for electron microscopy; Fassel TA et al.; Conventional methods of chemical fixation are often inadequate for preserving yeast ultrastructure . The thick cell wall severely limits penetration of fixatives rendering poor detail of the cell wall, membranes, and overall anatomy . Dimethylsulfoxide (DMSO) enhances penetration of chemicals and has been added to fixatives to improve cell preservation . At high concentrations (5 to 50%), however, it affects ultrastructure unpredictably . We found that adding 0.1% DMSO to fixatives greatly improved retention of yeast ultrastructure . Candida albicans, C . glabrata and Aspergillus fumigatus were fixed for 3 hr in 3% paraformaldehyde, 1% glutaraldehyde, 1 mM MgCl2, 1 mM CaCl2, 0.1% DMSO in 0.1 M sodium cacodylate buffer followed by 1% OsO4, 1% K2Cr2O7, 0.85% NaCl, 0.1% DMSO in the same buffer . Thin epoxy sections were post-stained in uranyl acetate and lead citrate . The multilayered character of the cell wall was distinct and well structured . Addition of ruthenium red or alcian blue to the fixatives further enhanced the outer fibrillar layer . The plasma membrane was contiguous and tightly adjacent to the inner mannoprotein layer of the cell wall . The cytoplasm was well preserved and the overall preservation of the yeast ultrastructure was significantly improved.

Mycoses, 1997 Sep, 40(3-4), 139 - 41
Sulconazole in the therapy of dermatomycoses in Nigeria; Gugnani HC et al.; A 1% cream of sulconazole nitrate, an imidazole derivative, was used to treat 38 patients with diverse clinical types of dermatomycoses, including 16 cases of pityriasis versicolor, 14 of dermatophytosis (tinea pedis, tinea cruris, tinea corporis), two of balanoposthitis due to Candida albicans, another two of candidosis of the groin, one each of groin and foot infection due to Trichosporon beigelii and one case each of lesions of the hand and trunk caused by Petriellidium boydii and Scytalidium hyalinum respectively . A complete cure was achieved in 91% of patients, with resolution of the lesions in the majority within 2-4 weeks . There were only two relapses . Sulconazole is recommended as an effective drug for topical treatment of superficial fungal infections of the skin.

Mycoses, 1997 Sep, 40(3-4), 119 - 25
Azole antifungals: weak inhibitors of inducible nitric oxide synthase in mouse and human cells; Vermuyten K et al.; The effect of three azole antifungals on inducible nitric oxide (iNOS) activity in different mouse and human cells was evaluated . The iNOS activity was determined by L-citrulline and nitrite measurement . In the murine macrophage cell line RAW 264.7, in mouse peritoneal macrophages (MPM) and in human colorectal adenocarcinoma cells (DLD-1), iNOS activity could be induced with lipopolysaccharides and cytokines . Under similar conditions, no iNOS induction was found in human monocytes/macrophages . The concentration of itraconazole, ketoconazole or miconazole needed to inhibit iNOS activity by 50% in RAW 264.7 cells, MPM and DLD-1 cells was > or = 10 mumol l-1 . This is at least 100 times more than the concentrations of these azole antifungals required to produce a 50% inhibition of yeast growth and ergosterol synthesis of, for example, Candida albicans after the same incubation period . These results show that azole antifungals are weak inhibitors of iNOS in intact cells.

J Biol Chem, 1997 Nov 28, 272(48), 30289 - 98
X-ray crystallographic studies of Candida albicans dihydrofolate reductase . High resolution structures of the holoenzyme and an inhibited ternary complex; Whitlow M et al.; The recent rise in systemic fungal infections has created a need for the development of new antifungal agents . As part of an effort to provide therapeutically effective inhibitors of fungal dihydrofolate reductase (DHFR), we have cloned, expressed, purified, crystallized, and determined the three-dimensional structure of Candida albicans DHFR . The 192-residue enzyme, which was expressed in Escherichia coli and purified by methotrexate affinity and cation exchange chromatography, was 27% identical to human DHFR . Crystals of C . albicans DHFR were grown as the holoenzyme complex and as a ternary complex containing a pyrroloquinazoline inhibitor . Both complexes crystallized with two molecules in the asymmetric unit in space group P21 . The final structures had R-factors of 0.199 at 1.85-A resolution and 0.155 at 1.60-A resolution, respectively . The enzyme fold was similar to that of bacterial and vertebrate DHFR, and the binding of a nonselective diaminopyrroloquinazoline inhibitor and the interactions of NADPH with protein were typical of ligand binding to other DHFRs . However, the width of the active site cleft of C . albicans DHFR was significantly larger than that of the human enzyme, providing a basis for the design of potentially selective inhibitors.

Blood, 1997 Dec 1, 90(11), 4485 - 92
FcalphaRI (CD89) as a novel trigger molecule for bispecific antibody therapy; Valerius T et al.; Promising results from clinical trials with unconjugated antibodies stimulated renewed interest in immune effector mechanisms of monoclonal antibodies (MoAbs) . We investigated the potential of IgA as antibody isotype for cell- or complement-mediated tumor cell lysis and assessed the potential of its myeloid Fc receptor, FcalphaRI (CD89), as trigger molecule for bispecific antibody (BsAb)-mediated immunotherapy . Comparing hapten-directed antibodies of human IgA2 with IgG1 or IgG3 isotypes, we found all three to mediate effective killing of sensitized tumor target cells in whole blood assays . Analysis of effector mechanisms showed IgG-mediated lysis to be predominantly complement-dependent, whereas IgA-dependent killing was primarily effector cell-mediated . A comparison of effector cell populations in antibody-dependent cell-mediated cytotoxicity (ADCC) showed neutrophils to be most important for IgA-dependent tumor cell killing, involving FcalphaRI as shown with Fc receptor blocking antibodies . Reverse ADCC experiments against target cells sensitized with Fc receptor antibodies, or assays with FcalphaRI-directed bispecific antibodies confirmed FcalphaRI as effective trigger molecule in polymorphonuclear neutrophil (PMN)-mediated lysis . During granulocyte colony-stimulating factor (G-CSF ) therapy, (FcalphaRI x HER-2/neu) bispecific antibodies induced enhanced killing of HER-2/neu positive SK-BR-3 breast cancer cells in whole blood assays . This enhanced cytotoxicity was paralleled by increased PMN counts, which lead to higher effector to target cell ratios in G-CSF-primed blood . Furthermore, bispecific antibodies, directed to FcalphaRI and Candida albicans, enhanced neutrophils' phagocytosis of fungi . In summary, these results identify IgA as an effective antibody isotype for immunotherapy, working primarily via FcalphaRI on neutrophils . They suggest FcalphaRI-directed bispecific antibodies and G-CSF to be an attractive combination for malignant or infectious diseases.

J Oral Rehabil, 1997 Oct, 24(10), 788 - 90
Existence of Candida albicans and microorganisms in denture stomatitis patients; Kulak Y et al.; The aetiology of denture stomatitis is not clear from the literature . Some studies show its aetiology as Candida albicans, while other reports point out the significance of microorganisms . In this study the existence of C . albicans and microorganisms was investigated in subjects with and without denture stomatitis . The results showed that a combination of C . albicans and microorganisms is more likely to be responsible for denture stomatitis.

J Antimicrob Chemother, 1997 Oct, 40(4), 511 - 6
Fluconazole disc diffusion testing for the routine laboratory; May JL et al.; The increasing incidence of resistance to the antifungal agent, fluconazole, prompted the need for a rapid, reliable and easy-to-use susceptibility test . We have developed a disc diffusion test for fluconazole against Candida spp . suitable for a clinical laboratory . Disc diffusion tests on six different media were compared with MIC values . On the basis of correlation coefficient with MICs (r = -0.95), quality of growth and zone edge definition, Yeast Nitrogen Base agar with glucose (YNBG) produced the best results . Further studies on YNBG showed that the method is reliable for Candida albicans and for resistant isolates with no zone of inhibition, but results for the slower growing and uncommon species must be interpreted with some caution . Implementation of this test in the clinical laboratory has provided a much needed therapeutic service for clinicians within the hospital . It has also reduced the reliance on the reference laboratory for susceptibility results and the consequent costs involved.

J Ethnopharmacol, 1997 Oct, 58(2), 75 - 83
Antimicrobial screening of selected medicinal plants from India; Valsaraj R et al.; From the Indian traditional medicines 78 plants were selected on the basis of their use in the treatment of infectious diseases . Different concentrations of 80% ethanol extracts were tested, using the agar dilution method, against four bacteria: Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa and, using the agar-well diffusion method, against two fungi: Candida albicans and Aspergillus niger . In the lowest tested concentration of 1.6 mg/ml, 10% of the plant extracts were active; 44% in a concentration of 6.25 mg/ml and 90% of the plant extracts were active against at least two bacteria in a concentration of 25 mg/ml . Only 13% of the plant extracts were active against at least one fungus in a concentration of 50 mg/ml.

Mycopathologia, 1997, 138(1), 29 - 35
Effect of iron on fluconazole activity against Candida albicans in presence of human serum or monocyte-derived macrophages; Minn Y et al.; Human serum, transferrin, and apotransferrin are known to profoundly inhibit the growth of Candida albicans by iron deprivation . On the other hand, iron overload (iron saturated transferrin) is a serious risk factor for candidiasis in newborn and in leukemic patients . We tested the efficacy of fluconazole and the previously demonstrated synergy of fluconazole and effector cells against C . albicans under iron overload conditions where efficacy might be diminished . We confirm that exogenous iron completely reversed the inhibitory effect of human serum and report that the efficacy of fluconazole against C . albicans was not significantly compromised in a 24 h assay system . Although exogenous iron inhibited fungistatic activity of monocyte-derived macrophages, it did not interfere with the synergistic candidacidal activity of fluconazole and monocyte-derived macrophages . In 72 h assays, where fluconazole had candidacidal activity, exogenous iron did not compromise efficacy of fluconazole, and fluconazole activity was often increased . These in vitro results suggest that effectiveness of fluconazole therapy would not be compromised in iron overload situations in vivo.

Mycopathologia, 1997, 138(1), 21 - 8
Protein A induced protection against experimental candidiasis in mice; Srivastava AK et al.; Staphylococcus aureus protein-A (SpA), an important multipotent immunostimulator, increased the resistance to infection with Candida albicans in Swiss albino mice . The mice treated with repeated immunologically active doses (1 microgram/mouse) of SpA, pre- and post-infection, showed one hundred percent protection against experimental candidiasis, and constituted the first such report . Protection could be demonstrated on the basis of leukocyte counts, colony forming unit (CPU) kinetics in liver, spleen, kidney and peritoneal cavity, and phagocytosis by monocyte-derived macrophages.

Eur J Clin Pharmacol, 1997, 53(2), 101 - 4
Povidone-iodine wash solutions in the prevention of superficial fungal infections; predictive evaluation using the corneofungimetry bioassay; Pierard GE et al.; OBJECTIVE: Prevention of superficial mycoses remains a stubborn problem . The effect of antiseptics for that purpose is largely unknown . We studied the potential fungitoxic activity of povidone iodine (PVP-I) contained in wash solutions . METHODS: The corneofungimetry bioassay was conducted using PVP-I at 1.33%, 2.5%, 4% and 7.5% as test products and four target fungi, namely Candida albicans, Trichophyton rubrum, T . mentagrophytes, var . interdigitale (20 strains) and Microsporum canis . RESULTS: Data show that PVP-I limits fungal growth on stratum corneum . Different species and strains of fungi are not similarly affected . There also exists a diversity of individual susceptibility of the stratum corneum to promote germination of yeasts and arthroconidia.

Dermatology, 1997, 195 Suppl 2, 111 - 6
Molecular effects of povidone-iodine on relevant microorganisms: an electron-microscopic and biochemical study; Schreier H et al.; The aim of this study was to elucidate the effects of povidone-iodine (PVP-I) on cell ultrastructure by electron microscopy and to monitor changes in enzyme activity and nucleotide efflux . Staphylococcus aureus, Escherichia coli and Candida albicans, medically relevant gram-positive, gram-negative and yeast microorganisms, served as models . In the presence of PVP-I, rapid partitioning of the cytoplasm and pronounced coagulation of nuclear material was noted . E . coli and S . aureus showed no major structural wall damage . C . albicans exhibited a rapid, dose-dependent 'loosening' of the cell wall; cells remained intact without lysis, rupture or wall breakage . Changes in beta-galactosidase and nucleotide concentrations were measured in E . coli . A rapid and dose-dependent loss of cellular beta-galactosidase activity was found, with no increase in the supernatant; loss of cellular nucleotides corresponded with an increase in the supernatant . Electron-microscopic and biochemical observations support the conclusion that PVP-I interacts with cell walls of microorganisms causing pore formation or generating solid-liquid interfaces at the lipid membrane level which lead to loss of cytosol material, in addition to enzyme denaturation.

J Med Vet Mycol, 1997 Sep-Oct, 35(5), 305 - 12
Characterization of the phosphoribosylpyrophosphate synthetase gene from Candida albicans; Payne T et al.; Phosphoribosylpyrophosphate (PRPP) synthetase catalyzes the transfer of phosphate from ATP to D-ribose-5-phosphate during the synthesis of purine and pyrimidine nucleotides and tryptophan and histidine biosynthesis in a variety of organisms . We cloned and sequenced the PRPP synthetase gene (PRS1) of Candida albicans because, in Saccharomyces cerevisiae, a deficiency in PRPP synthetase activity interacts with a mutation in ELM4-1 (elongated morphology) to cause constitutive pseudohyphal growth in nitrogen-rich media . In order to study the role of the C . albicans PRS1 in growth and morphogenesis, we used gene disruption to isolate PRS1 mutants; however, while heterozygous PRS1 clones were readily obtained, homozygous, null strains were not recovered indicating that PRS1 is probably essential for growth of the organism . Heterozygotes in PRS1 produced approximately 35% less PRPP synthetase (P = 0.0004) and exhibited a similar reduction in transcript levels . Confirmation of a heterozygous, single disruption in PRS1 was obtained by I-SceI digestion of chromosomal-sized DNA and Southern blot hybridizations . While no role in morphogenesis is elucidated by this work, the data strongly suggests that PRS1 is an essential gene in C . albicans and supports earlier results that indicated the presence of a single PRS gene in C . albicans.

J Bacteriol, 1997 Dec, 179(24), 7734 - 41
Cloning of the RHO1 gene from Candida albicans and its regulation of beta-1,3-glucan synthesis; Kondoh O et al.; The Saccharomyces cerevisiae RHO1 gene encodes a low-molecular-weight GTPase . One of its recently identified functions is the regulation of beta-1,3-glucan synthase, which synthesizes the main component of the fungal cell wall (J . Drgonova et al., Science 272:277-279, 1996; T . Mazur and W . Baginsky, J . Biol . Chem . 271:14604-14609, 1996; and H . Qadota et al., Science 272:279-281, 1996) . From the opportunistic pathogenic fungus Candida albicans, we cloned the RHO1 gene by the PCR and cross-hybridization methods . Sequence analysis revealed that the Candida RHO1 gene has a 597-nucleotide region which encodes a putative 22.0-kDa peptide . The deduced amino acid sequence predicts that Candida albicans Rho1p is 82.9% identical to Saccharomyces Rho1p and contains all the domains conserved among Rho-type GTPases from other organisms . The Candida albicans RHO1 gene could rescue a S . cerevisiae strain containing a rho1 deletion . Furthermore, recombinant Candida albicans Rho1p could reactivate the beta-1,3-glucan synthesis activities of both C . albicans and S . cerevisiae membranes in which endogenous Rho1p had been depleted by Tergitol NP-40-NaCl treatment . Candida albicans Rho1p was copurified with the beta-1,3-glucan synthase putative catalytic subunit, Candida albicans Gsc1p, by product entrapment . Candida albicans Rho1p was shown to interact directly with Candida albicans Gsc1p in a ligand overlay assay and a cross-linking study . These results indicate that Candida albicans Rho1p acts in the same manner as Saccharomyces cerevisiae Rho1p to regulate beta-1,3-glucan synthesis.

Mol Biol Cell, 1997 Dec, 8(12), 2539 - 51
Derepressed hyphal growth and reduced virulence in a VH1 family-related protein phosphatase mutant of the human pathogen Candida albicans; Csank C et al.; Mitogen-activated protein (MAP) kinases are pivotal components of eukaryotic signaling cascades . Phosphorylation of tyrosine and threonine residues activates MAP kinases, but either dual-specificity or monospecificity phosphatases can inactivate them . The Candida albicans CPP1 gene, a structural member of the VH1 family of dual- specificity phosphatases, was previously cloned by its ability to block the pheromone response MAP kinase cascade in Saccharomyces cerevisiae . Cpp1p inactivated mammalian MAP kinases in vitro and acted as a tyrosine-specific enzyme . In C . albicans a MAP kinase cascade can trigger the transition from the budding yeast form to a more invasive filamentous form . Disruption of the CPP1 gene in C . albicans derepressed the yeast to hyphal transition at ambient temperatures, on solid surfaces . A hyphal growth rate defect under physiological conditions in vitro was also observed and could explain a reduction in virulence associated with reduced fungal burden in the kidneys seen in a systemic mouse model . A hyper-hyphal pathway may thus have some detrimental effects on C . albicans cells . Disruption of the MAP kinase homologue CEK1 suppressed the morphological effects of the CPP1 disruption in C . albicans . The results presented here demonstrate the biological importance of a tyrosine phosphatase in cell-fate decisions and virulence in C . albicans.

J Clin Microbiol, 1997 Dec, 35(12), 3311 - 2
Comparison of different methods for extraction of DNA of fungal pathogens from cultures and blood; Loffler J et al.; Five commercially available extraction kits and an in-house DNA extraction method for the release of DNA from Candida albicans and Aspergillus niger cells were assessed for sensitivity, purity, duration, and cost . All commercially available kits helped to shorten the duration of DNA extraction . However, the sensitivity varied from 1 to 1,000 fungal cells/ml and costs varied from $0.10 to 2.30 . The QIAmp Tissue kit was the commercially available assay that yielded the same sensitivity and purity of fungal DNA release as the in-house protocol but was the most expensive method . In comparing these two extraction protocols, a 99% concordance of PCR results for 125 blood samples analyzed could be demonstrated.

J Clin Microbiol, 1997 Dec, 35(12), 3284 - 7
Detection of antibodies to Candida albicans germ tubes for diagnosis and therapeutic monitoring of invasive candidiasis in patients with hematologic malignancies; Garcia-Ruiz JC et al.; We prospectively investigated the ability of detection of antibodies to Candida albicans germ tubes (CAGT) to diagnose invasive candidiasis in 95 consecutive admissions of 73 patients with hematologic disorders undergoing intensive chemotherapy . The episodes were divided into three groups according to clinical and microbiological diagnosis . Group 1 comprised eight admissions of eight patients with invasive candidiasis . Group 2 comprised 42 admissions of 34 patients without evidence of invasive candidiasis . Group 3 comprised the remaining 45 admissions of 37 patients with febrile episodes which were not diagnosed by microbiological culture . Antibodies to CAGT were detected in 87.5% of group 1 patients . Detection of antibodies to CAGT in patients with Candida fungemia was delayed somewhat relative to the time the blood culture was positive, but antibodies to CAGT were detected earlier than a diagnosis was made in patients with deep-tissue candidiasis . Sera from 2 admissions in group 2 and 12 admissions in group 3 revealed antibodies to CAGT . At a titer of > or = 1:20, detection of antibodies to CAGT had a sensitivity of 87.5%, specificity of 95.2%, positive predictive value of 77.8%, and negative predictive value of 97.6% . Antibodies to CAGT were usually detected before beginning of empiric antifungal therapy . Titers of antibodies to CAGT were maintained in most patients who died but declined and eventually disappeared in the patients who survived . Since antibodies to CAGT were detected in all patients with tissue-proven invasive candidiasis but negative by blood culture, detection of antibodies to CAGT complemented blood cultures for diagnosis and therapeutic monitoring of patients with hematologic malignancies and invasive candidiasis.

J Clin Microbiol, 1997 Dec, 35(12), 3032 - 6
Molecular tracking of Candida albicans in a neonatal intensive care unit: long-term colonizations versus catheter-related infections; Ruiz-Diez B et al.; Nosocomial neonatal candidiasis is a major problem in infants requiring intensive therapy . The subjects of this retrospective study were nine preterm infants admitted to the neonatal intensive care unit of the Hospital Central de Asturias between March 1993 and August 1994 . The infants were infected with or colonized by Candida albicans . Five patients developed C . albicans bloodstream infections . A total of 36 isolates (including isolates from catheters and parenteral nutrition) were examined for molecular relatedness by PCR fingerprinting and restriction fragment length polymorphism (RFLP) analysis . The core sequence of phage M13 was used as a single primer in the PCR-based fingerprinting procedure, and RFLP analysis was performed with C . albicans-specific DNA probe 27A . Both techniques were evaluated with a panel of eight C . albicans reference strains, and each technique showed eight different patterns . With the 36 isolates from neonates, each technique enabled us to identify by PCR and RFLP analysis seven and six different patterns, respectively . The combination of these two methods (composite DNA type) identified eight different profiles . A strain with one of these profiles was present in three patients and in their respective catheters . Patients infected with or colonized by this isolate profile were clustered in time . Among the other patients, each patient was infected over time and at multiple anatomic sites with a C . albicans strain with a distinct DNA type . We conclude that C . albicans was most commonly producing long-term colonizations, although horizontal transmission probably due to catheters also occurred.

Phytochemistry, 1996 Jul, 42(5), 1315 - 20
Antifungal and antibacterial naphthoquinones from Newbouldia laevis roots; Gafner S et al.; From a dichloromethane extract of Newbouldia laevis roots, four new (6-hydroxydehydroiso-alpha-lapachone, 7-hydroxydehydroiso-alpha-lapachone, 5,7-dihydroxydehydroiso-alpha-lapachone and 3-hydroxy-5-methoxydehydroiso-alpha-lapachone) and six known naphthoquinones have been isolated . Their structures were established by spectroscopic methods (UV, EI mass spectrometry, 1H and 13C NMR) and that of 7-hydroxydehydroiso-alpha-lapachone was confirmed by X-ray crystallography . All naphthoquinones showed antifungal activity against Cladosporium cucumerinum and Candida albicans, and activity against the bacteria Bacillus subtilis and Escherichia coli.

Phytochemistry, 1996 Jul, 42(5), 1305 - 13
Acyl secoiridoids and antifungal constituents from Gentiana macrophylla; Tan RX et al.; LC-UV-mass spectrometry and bioassay co-directed fractionation of an aqueous acetone extract of the roots of Gentiana macrophylla gave three new chromene derivatives and two novel and six known secoiridoids, along with kurarinone, kushenol I, beta-sitosterol, stigmasterol, daucosterol, beta-sitosterol-3-O-gentiobioside, alpha-amyrin, oleanolic acid, isovitexin, gentiobiose and methyl 2-hydroxy-3-(1-beta-D-glucopyranosyl)oxybenzoate . The structures of the new products were established from spectral and chemical evidence as 2-methoxyanofinic acid and macrophyllosides A-D . The six known secoiridoids were gentiopicroside, sweroside, 6'-O-beta-D-glucosylgentiopicroside, 6'-O-beta-D-glucosylsweroside, trifloroside and rindoside . The new acid (2-methoxyanofinic acid), its methyl ester, kurarinone and kushenol I were shown to be active against the plant pathogenic fungus Cladosporium cucumerinum . The methyl ester and kurarinone inhibited also the growth of the human pathogenic yeast Candida albicans . Structure-activity relationships were studied . Thus, addition of a methoxyl group to the benzene nucleus of anofinic acid (2,2-dimethyl-2H-1-benzopyran-6-carboxylic acid) increased the antifungal activity remarkably whereas glycosylation at the carboxylic moiety was found to remove the activity . Esterification of the new acid induced its activity against C . albicans, but decreased its growth inhibition properties against C . cucumerinum . Hydroxylation of kurarinone at the 3 beta-position removed its activity against C . albicans and decreased the inhibition of C . cucumerinum . In addition, the chemotaxonomic significance of the identified constituents is discussed.

J Infect Dis, 1997 Dec, 176(6), 1579 - 83
Aspergillus fumigatus conidia induce a Th1-type cytokine response; Grazziutti ML et al.; The response of human peripheral blood mononuclear cells (MNC) to Aspergillus fumigatus in vitro was evaluated . In studies of the proliferative response of MNC from 18 healthy donors to heat-killed A . fumigatus conidia, 15 displayed a significant response, with a stimulation index (SI) between 4 and 193 . In contrast, all donors displayed a positive response to Candida albicans blastoconidia (SI ranged from 10 to 224) . Despite the variability in reactivity to A . fumigatus conidia, the response of a particular individual was stable when retested over periods of 1-2 weeks . Supernatant from cocultures of A . fumigatus conidia with MNC contained increased levels of interferon-gamma, granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha, and interleukin (IL)-2, compared with unstimulated cells, but not IL-10 or IL-4 . In addition, A . fumigatus induced lymphocyte surface expression of adhesion/activation-associated molecules . These results suggest that lymphocytes may contribute to host defense against Aspergillus by generating a Th1-type response.

J Infect Dis, 1997 Dec, 176(6), 1567 - 78
Mechanisms and target sites of damage in killing of Candida albicans hyphae by human polymorphonuclear neutrophils; Christin L et al.; Target sites of fungal cell damage were studied to define mechanisms of neutrophil-mediated killing of Candida albicans hyphae . Neutrophils induced hyphal cell wall damage, as evidenced by release of cell wall glycoproteins and confocal microscopic changes . Damage occurred in the presence of neutrophil granule extracts and did not require oxidants . However, oxidation of hyphal surface glycoproteins correlated strongly with parallel increments in fungicidal activity, suggesting that oxidants did contribute to maximal cell wall damage . Neutrophil oxidants also induced hyphal DNA fragmentation, primarily single-strand breakage, as shown by increased electrophoretic migration after nuclease-S1 DNA digestion at single-strand break sites . The onset of damage to hyphal cell walls and DNA preceded detectable neutrophil-mediated fungicidal effects . Likewise, hyphal amino acid and nucleotide turnover as well as ATP initially rose, then declined as lethal effects became detectable . Thus, preceding detectable fungal cell death, neutrophil oxidative and oxygen-independent mechanisms damaged defined targets.

FEMS Immunol Med Microbiol, 1997 Oct, 19(2), 169 - 80
Interleukin-4 suppresses antifungal activity of human mononuclear phagocytes against Candida albicans in association with decreased uptake of blastoconidia; Roilides E et al.; Pathogenesis of invasive candidiasis may involve regulatory activities of Th2 immunity on phagocytic host defenses . The effects of interleukin (IL)-4 on antifungal capacity of human mononuclear phagocytes against Candida albicans were studied . Incubation of adherent mononuclear leukocytes from healthy donors with IL-4 (1-5 ng ml(-1)) at 37 degrees C for 2-4 days suppressed uptake of C . albicans blastoconidia in the presence of human serum (P < or = 0.01), and anti-IL-4 inhibited its suppressive effect . The effect of IL-4 was protein synthesis-dependent . Interferon-gamma (0.25-25 ng ml(-1)), granulocyte-macrophage colony-stimulating factor (CSF, 20 ng ml(-1)), macrophage-CSF (15 ng ml(-1)) but not IL-10 (100 ng ml(-1)) somewhat counteracted the suppressive effect of IL-4 . In contrast, mannose receptor-mediated uptake of blastoconidia in the absence of serum was increased by IL-4 . Killing of conidia was decreased after incubation of morphonuclear leukocytes with IL-4 for 2 days (P < 0.05) . While superoxide anion production in response to phorbol myristate acetate was decreased by IL-4 (P < 0.05), it was not altered in response to blastoconidia and pseudohyphae . Morphonuclear leukocyte-induced pseudohyphal damage also remained unaltered . These findings suggest that IL-4 plays its detrimental role in invasive candidiasis by predominantly suppressing uptake and killing of blastoconidia by morphonuclear leukocytes . Anti-IL-4, IFN-gamma, GM-CSF and M-CSF appear to counteract suppression of morphonuclear leukocyte phagocytic activity suggesting new approaches to the management of disseminated candidiasis.

Jpn J Antibiot, 1997 Sep, 50(9), 799 - 805
{The in vitro antifungal activities of fluconazole against pathogenic yeasts recently isolated from clinical specimens}; Yamaguchi H et al.; The emergence of Candida albicans resistance to azole antifungal agents have been reported in the U . S . and Europe . We examined the in vitro antifungal activities of fluconazole against clinical isolates collected by seven investigators in three years to examine if a tendency existed toward the development of azole-resistance among fungal isolates in Japan . The following results were obtained: 1 . Sensitivities to fluconazole (FLCZ) were determined for yeast-like fungi, including 113 strains isolated in 1993, 149 strains isolated in 1994 and 205 strains isolated in 1995 . No significant differences in sensitivities in the three years were detected . 2 . Minimum inhibitory concentrations of FLCZ were 0.1-0.78 microgram/ml for C . albicans and 3.13-25 micrograms/ml for C . glabrata . Strains with 25 micrograms/ml of FLCZ's MIC were detected; two strains of C . krusei and one strain each of C . krusei, Trichospron beigelii and Hansenula anomala . No strains with higher than 50 micrograms/ml MIC of FLCZ were detected . 3 . In vitro activities of FLCZ were compared between clinical strains isolated between 1993 and 1995 and clinical strains isolated before the marketing of FLCZ (up to December 1987) or clinical yeasts isolated between 1991 and 1992 . No significant differences were observed, suggesting that no tendency existed toward azole resistance among fungal strains examined.

Infect Immun, 1997 Dec, 65(12), 5354 - 7
A monoclonal antibody to Candida albicans enhances mouse neutrophil candidacidal activity; Caesar-TonThat TC et al.; A monoclonal antibody (MAb) to Candida albicans (MAb B6.1) that protects against candidiasis and the nonprotective MAb B6 were compared for ability to support neutrophil (polymorphonuclear leukocyte {PMN}) candidacidal activity . Both MAbs are immunoglobulin M, and each recognizes distinct C . albicans mannan cell wall determinants . PMN candidacidal activity was assessed by transmission electron microscopy and by an in vitro killing assay . The results indicated that MAb B6.1, but not MAb B6, enhances ingestion and killing of yeast cells by PMN in the presence of serum complement.

Infect Immun, 1997 Dec, 65(12), 5289 - 94
Expression, cloning, and characterization of a Candida albicans gene, ALA1, that confers adherence properties upon Saccharomyces cerevisiae for extracellular matrix proteins; Gaur NK et al.; Adherence of Candida albicans to host tissues is a necessary step for maintenance of its commensal status and is likely a necessary step in the pathogenesis of candidiasis . The extracellular matrix (ECM) proteins are some of the host tissue and plasma proteins to which C . albicans adheres through adhesins located on the fungal cell surface . To isolate genes encoding ECM adhesins, an assay was developed based on the ability of yeast cells to adhere to magnetic beads coated with the ECM protein fibronectin, type IV collagen, or laminin . A C . albicans genomic library was constructed by cloning XbaI-partially-digested and size-selected fragments into pAUR112, an Escherichia coli-yeast low-copy-number shuttle vector . The C . albicans library was transformed into Saccharomyces cerevisiae YPH 499, and clones capable of adherence were selected by using ECM protein-coated magnetic beads . A plasmid containing an approximately 8-kb insert was isolated from 29 adherent clones . These clones exhibited adherence to all ECM protein-coated magnetic beads and to human buccal epithelial cells . The ALA1 gene (for agglutinin-like adhesin) was localized by subcloning it into a 5-kb XbaI fragment which retained the adherence phenotype in both orientations . The complete DNA sequence of the 5-kb insert was determined, and an open reading frame (ORF) encoding 1,419 amino acid residues was identified . Deletions from the 5' and 3' ends extending into the DNA sequence encoding the 1,419-amino-acid ORF product inactivated the adherence phenotype, suggesting that it is the coding region of the ALA1 gene . A database search identified ALA1 to be similar to the C . albicans ALS1 (for agglutinin-like sequence 1) protein and the S . cerevisiae agglutinin protein (AG alpha1), although the homology at the primary amino acid sequence level is limited to the first half of each of these proteins . ALA1 contains a central domain of six tandem repeats of 36 amino acids . We discuss the significance of various predicted ALA1 structural motifs and their relationships to function in the adherence process.

Infect Immun, 1997 Dec, 65(12), 4909 - 17
A gene homologous to Saccharomyces cerevisiae SNF1 appears to be essential for the viability of Candida albicans; Petter R et al.; The SNF1 gene of Saccharomyces cerevisiae (ScSNF1) is essential for the derepression of catabolic repression . We report here the isolation and characterization of an SNF1 homolog from Candida albicans (CaSNF1) which is apparently essential for the viability of this organism . The putative amino acid sequence of CaSNF1 has 68% identity with that of ScSNF1 and can restore the S . cerevisiae snf1 delta mutant's ability to utilize sucrose . Disruption of one of the CaSNF1 alleles resulted in morphological changes and decreased growth rates but did not modify the carbon source utilization pattern . Repetitive unsuccessful attempts to generate a snf1/snf1 homozygote by disruption of the second allele, using various vectors and approaches, suggest the lethal nature of this mutation . Integration into the second allele was possible only when a full-length functional SNF1 sequence was reassembled, further supporting this hypothesis and indicating that the indispensability of Snf1p prevented the isolation of snf1/snf1 mutants . The mutant bearing two disrupted SNF1 alleles and the SNF1 functional sequence maintained its ability to utilize sucrose and produced stellate colonies with extensive hyphal growth on agar media . It was demonstrated that in a mouse model, the virulences of this mutant and the wild-type strain are similar, suggesting that hyphal growth in vitro is not an indicator for higher virulence.

J Bacteriol, 1997 Dec, 179(23), 7210 - 8
The Candida albicans CDR3 gene codes for an opaque-phase ABC transporter; Balan I et al.; We report the cloning and functional analysis of a third member of the CDR gene family in Candida albicans, named CDR3 . This gene codes for an ABC (ATP-binding cassette) transporter of 1,501 amino acids highly homologous to Cdr1p and Cdr2p (56 and 55% amino acid sequence identity, respectively), two transporters involved in fluconazole resistance in C . albicans . The predicted structure of Cdr3p is typical of the PDR/CDR family, with two similar halves, each comprising an N-terminal hydrophilic domain with consensus sequences for ATP binding and a C-terminal hydrophobic domain with six predicted transmembrane segments . Northern analysis showed that CDR3 expression is regulated in a cell-type-specific manner, with low levels of CDR3 mRNA in CAI4 yeast and hyphal cells, high levels in WO-1 opaque cells, and undetectable levels in WO-1 white cells . Disruption of both alleles of CDR3 in CAI4 resulted in no obvious changes in cell morphology, growth rate, or susceptibility to fluconazole . Overexpression of Cdr3p in C . albicans did not result in increased cellular resistance to fluconazole, cycloheximide, and 4-nitroquinoline-N-oxide, which are known substrates for different transporters of the PDR/CDR family . These results indicate that despite a high degree of sequence conservation with C . albicans Cdr1p and Cdr2p, Cdr3p does not appear to be involved in drug resistance, at least to the compounds tested which include the clinically relevant antifungal agent fluconazole . Rather, the high level of Cdr3p expression in WO-1 opaque cells suggests an opaque-phase-associated biological function which remains to be identified.

APMIS, 1997 Nov, 105(11), 875 - 83
The antifungal effect of lactoferrin and lysozyme on Candida krusei and Candida albicans; Samaranayake YH et al.; Lactoferrin and lysozyme (muramidase) are non-immune defence factors present in various exocrine secretions, including saliva . Previous studies have shown that both proteins, either singly or in combination, are bactericidal in nature and their combined activity is synergistic . As little is known of their interactions with Candida species, 20 oral isolates of C . krusei and 5 isolates of C . albicans were studied for their susceptibility to human apo-lactoferrin and lysozyme, either singly or in combination, using an in vitro assay system . The two species exhibited significant interspecies differences in susceptibility to lactoferrin (p < 0.05), but not for lysozyme; C . krusei being more sensitive to lactoferrin (c 1.4 times) than C . albicans . Both species revealed significant intraspecies differences in their susceptibility to lysozyme (p < 0.05), but not for lactoferrin . No synergistic antifungal activity of the two proteins on either Candida species was noted . The results imply that the variable expression of the fungicidal activity of lactoferrin and lysozyme on Candida species may modulate the oral carriage of yeasts in a complex manner.

Yeast, 1997 Nov, 13(14), 1383 - 9
Isolation and sequence of the gene encoding ornithine decarboxylase, SPE1, from Candida albicans by complementation of a spe1 delta strain of Saccharomyces cerevisiae; McNemar MD et al.; The gene encoding ornithine decarboxylase, SPE1, from the pathogenic yeast Candida albicans has been isolated by complementation of an ornithine decarboxylase-negative (spe1 delta) strain of Saccharomyces cerevisiae . Four transformants, three of which contain plasmids with the SPE1 gene, were isolated by selection on polyamine-free medium . The C . albicans ornithine decarboxylase (ODC) showed high homology with other eukaryotic ODCs at both the amino acid and nucleic acid levels.

Yeast, 1997 Nov, 13(14), 1375 - 81
Isolation of a putative prolyl-tRNA synthetase (CaPRS) gene from Candida albicans; Sentandreu M et al.; We have isolated a 4.0-kb fragment from a genomic library of Candida albicans which contained two open reading frames (ORFs) . One of them is homologous to a prolyl-tRNA synthetase that catalyses the charging of a specific tRNA by proline (CaPRS) . A deduced sequence of 575 amino acids representing a polypeptide of 66.2 kDa was determined . A FASTA search indicated that the CaPRSp had an overall similarity of 54.4% with the product of a Saccharomyces cerevisiae ORF (YER087) and 43.8% with the prolyl-tRNA synthetase of Escherichia coli (COLIPRO) . Consensus Class II aminoacyl-tRNA synthetase sequences were identified by the PROSITE program . CaPRS was localized to chromosome R of the C . albicans genome and CaPRS DNA hybridized to a major RNA transcript of 1.7 kb under all conditions tested.

Microbiology, 1997 Nov, 143 ( Pt 11), 3527 - 35
Two transcripts, differing at their 3' ends, are produced from the Candida albicans SEC14 gene; Riggle PJ et al.; A search for Candida albicans mutants defective in filamentous growth led to the isolation of a mutant strain with an insertion mutation in the SEC14 gene . SEC14 encodes the phosphatidylinositol/phosphatidylcholine transfer protein, an essential protein in the yeast Saccharomyces cerevisiae . In the dimorphic yeast Yarrowia lipolytica, SEC14 is needed for growth only in the hyphal form and is not required for growth in the yeast form . However, unlike Y . lipolytica SEC14, C . albicans SEC14 is probably essential for growth . Northern blot analysis and PCR amplification of transcripts produced from the SEC14 gene demonstrated that two transcripts differing at their 3' ends were produced . The two transcripts may regulate the activity of SEC14 so that Sec14p can perform two functions in C . albicans . One function may be an essential function analogous to the function of Sec14p in S . cerevisiae and the second function may be important during filamentous growth, analogous to the function of Sec14p in Y . lipolytica.

Microbiology, 1997 Nov, 143 ( Pt 11), 3521 - 6
Phospholipase D activity is required for dimorphic transition in Candida albicans; McLain N et al.; Candida albicans is an opportunistic pathogen that causes significant morbidity and mortality in immunocompromised patients . In this report, the presence of a phospholipase D (PLD) activity in C . albicans, designated CaPLD1, is demonstrated . This is the first description of PLD activity in this organism . CaPLD1 activity was stimulated by inducers of dimorphic transition . Furthermore, transition was stimulated by the addition of exogenous PLD to cells . The addition of 1-propanol to the medium, which resulted in the production of phosphatidylpropanol by CaPLD1 at the expense of the usual product phosphatidic acid, delayed the yeast to hypha transition . These results suggest that CaPLD1 may be an important regulator of dimorphic transition in C . albicans.

J Bacteriol, 1997 Nov, 179(22), 7118 - 28
pCal, a highly unusual Ty1/copia retrotransposon from the pathogenic yeast Candida albicans; Matthews GD et al.; Retrotransposons are mobile genetic elements . They can transpose via the reverse transcription of mRNA into double-stranded DNA (dsDNA) followed by the insertion of this dsDNA into new sites within the host genome . The unintegrated, linear, dsDNA form of retrotransposons is usually very rare . We report here the isolation of a retrotransposon from Candida albicans which is unusual in this respect . This element, which we have named pCal, was first identified as a distinct band when uncut C . albicans DNA was examined on an agarose gel . Sequence analysis of the cloned element revealed that it is a retrotransposon belonging to the Ty1/copia group . It is estimated that pCal produces 50 to 100 free, linear, dsDNA copies of itself per cell . This is a much higher level of expression than even that of the system in which Ty1 is expressed behind the highly active GAL1 promoter on a high-copy-number plasmid (about 10 copies per cell) . Another unusual feature of pCal is that its Pol enzymes are likely to be expressed via the pseudoknot-assisted suppression of an upstream, in-phase stop codon, as has been shown for Moloney murine leukemia virus.

Antimicrob Agents Chemother, 1997 Nov, 41(11), 2576 - 8
In vitro kill curves of a new semisynthetic echinocandin, LY-303366, against fluconazole-sensitive and -resistant Candida species; Karlowsky JA et al.; In vitro killing by a new semisynthetic echinocandin, LY-303366, was characterized using clinical isolates of fluconazole-sensitive (Y58) and -resistant (Y180) Candida albicans as well as Candida glabrata (Y7) and Candida krusei (Y171) . The 24-h kill curves for Y58 and Y180 demonstrated dose-independent killing of between 1 and 2 log10 with LY-303366 at concentrations of 0.1, 1, 10, 50, 100, and 1,000 times the MIC . Regrowth did not occur at 24 h with either C . albicans isolate at the aforementioned LY-303366 concentrations . At their MICs, LY-303366 and amphotericin B produced similar killing kinetics in cultures of Y58, Y180, Y7, and Y171, while all cultures exposed to fluconazole at its MIC demonstrated stasis or growth over 24 h.

Antimicrob Agents Chemother, 1997 Nov, 41(11), 2518 - 21
Effectiveness of quinolone antibiotics in modulating the effects of antifungal drugs; Sugar AM et al.; Quinolone antibacterial drugs inhibit DNA gyrase, a type 2 topoisomerase . Since topoisomerases are present in eukaryotic cells, it was of interest to evaluate the antifungal activities of two clinically available quinolones, ciprofloxacin and trovafloxacin, alone and in combination with amphotericin B or fluconazole, in vitro against Candida albicans and in a murine model of invasive candidiasis . The in vitro activity of trovafloxacin was also tested against other yeasts and molds . In vitro, trovafloxacin exhibited no antifungal activity against any of the fungi (MIC, >250 microg/ml) . There was also no effect of the quinolone on the in vitro activity of either antifungal drug . Marked antifungal effects were seen, however, in the murine model of candidiasis . In all experiments, control mice infected intravenously with C . albicans were dead by day 24 . While either quinolone had minimal effects on survival of mice when used alone in oral doses of up to 40 mg/kg twice daily, the combination of the quinolone with fluconazole (40 or 80 mg/kg given twice daily by oral gavage) was more effective in prolonging survival than was fluconazole alone . Colony counts of kidneys on days 12 and 30 showed similar reductions in C . albicans recovered from mice treated with fluconazole with or without trovafloxacin or amphotericin B with or without trovafloxacin . Survival of mice treated with a suboptimal dose of amphotericin B (0.2 mg/kg/day) was also improved when trovafloxacin (40 mg/kg) given twice daily was included (0 versus 27%, respectively; P < 0.05) . While the mechanisms of action of the combination of trovafloxacin and amphotericin B or fluconazole are unclear, further work focused on fungal topoisomerase inhibition and the mechanism of the antifungal effect of quinolone antibacterial drugs is warranted.

Antimicrob Agents Chemother, 1997 Nov, 41(11), 2492 - 6
A new triazole, voriconazole (UK-109,496), blocks sterol biosynthesis in Candida albicans and Candida krusei; Sanati H et al.; Voriconazole (UK-109,496) is a novel triazole derivative with potent broad-spectrum activity against various fungi, including some that are inherently resistant to fluconazole, such as Candida krusei . In this study we compared the effect of subinhibitory concentrations of voriconazole and fluconazole on sterol biosynthesis of fluconazole-resistant and -susceptible Candida albicans strains, as well as C . krusei, in an effort to delineate the precise mode of action of voriconazole . Voriconazole MICs ranged from 0.003 to 4 microg/ml, while fluconazole MICs ranged from 0.25 to >64 microg/ml . To investigate the effects of voriconazole and fluconazole on candidal sterols, yeast cells were grown in the absence and presence of antifungals . In untreated C . albicans controls, ergosterol was the major sterol (accounting for 53.6% +/- 2.2% to 71.7% +/- 7.8% of the total) in C . albicans and C . krusei strains . There was no significant difference between the sterol compositions of the fluconazole-susceptible and -resistant C . albicans isolates . Voriconazole treatment led to a decrease in the total sterol content of both C . albicans strains tested . In contrast, exposure to fluconazole did not result in a significant reduction in the total sterol content of the three candidal strains tested (P > 0.5) . Gas-liquid chromatographic analysis revealed profound changes in the sterol profiles of both C . albicans strains and of C . krusei in response to voriconazole . This antifungal agent exerted a similar effect on the sterol compositions of both fluconazole-susceptible and -resistant C . albicans strains . Interestingly, a complete inhibition of ergosterol synthesis and accumulation of its biosynthetic precursors were observed in both strains treated with voriconazole . In contrast, fluconazole partially inhibited ergosterol synthesis . Analysis of sterols obtained from a fluconazole-resistant C . albicans strain grown in the presence of different concentrations of voriconazole showed that this agent inhibits ergosterol synthesis in a dose-dependent manner . In C . krusei, voriconazole significantly inhibited ergosterol synthesis (over 75% inhibition) . C . krusei cells treated with voriconazole accumulated the following biosynthetic intermediates: squalene, 4,14-dimethylzymosterol, and 24-methylenedihydrolanosterol . Accumulation of these methylated sterols is consistent with the premise that this agent functions by inhibiting fungal P-450-dependent 14alpha-demethylase . As expected, treating C . krusei with fluconazole minimally inhibited ergosterol synthesis . Importantly, our data indicate that voriconazole is more effective than fluconazole in blocking candidal sterol biosynthesis, consistent with the different antifungal potencies of these compounds.

Antimicrob Agents Chemother, 1997 Nov, 41(11), 2471 - 9
Identification of the FKS1 gene of Candida albicans as the essential target of 1,3-beta-D-glucan synthase inhibitors; Douglas CM et al.; Pneumocandins and echinocandins are fungicidal antibiotics, currently in clinical development, that inhibit 1,3-beta-D-glucan synthase (GS) in several human fungal pathogens . We have identified a gene from the diploid organism Candida albicans that encodes a target of these inhibitors . A 2.1-kb portion of this gene, designated CaFKS1, has significant homology to the Saccharomyces cerevisiae FKS1 and FKS2 genes, which encode partially functionally redundant subunits of GS . To evaluate the role of CaFkslp in susceptibility to echinocandins, we disrupted CaFKS1 on one homolog each of the spontaneous pneumocandin-resistant C . albicans mutants CAI4R1, NR2, NR3, and NR4 . These mutants had been selected previously on agar plates containing the pneumocandin L-733,560 . The clones derived from this transformation were either resistant (Ech{r}) or fully sensitive (Ech{s}) to inhibition by L-733,560 in both liquid broth microdilution and in vitro GS assays . The site of plasmid insertion in the transformants was mapped by Southern blot analysis, using restriction site polymorphisms in the CaFKS1 gene to distinguish between the two alleles (designated CaFKS1h and CaFKS1b) . For strains CAI4R1 and NR2, the CaFKS1b allele was disrupted in each Ech(r) transformant; for strain NR4, CaFKS1h was disrupted in each Ech(r) transformant . We conclude that (i) strains CAI4R1, NR2, and NR4 are heterozygous for a dominant or semidominant pneumocandin resistance mutation at CaFKS1, (ii) drug resistance mutations can occur in either CaFKS1 allele, and (iii) CaFks1p is a target of the echinocandins . For transformants of strain NR3, all the clones we analyzed were uniformly Ech(r), and only the CaFKS1h allele, either in disrupted or wild-type form, was detected on genomic Southern blots . We believe gene conversion at the CaFKS1 locus may have produced two Cafks1h alleles that each contain an Ech(r) mutation . Transformants derived from the mutants were analyzed for susceptibility to pneumocandin treatment in a mouse model of disseminated candidiasis . Strains heterozygous for the resistant allele (i.e., C . albicans CAI4R1, NR2, and NR4) were moderately resistant to treatment, while strains without a functional Ech(s) allele (i.e., strain NR3 and derivatives of strain CAI4R1 with the disruption plasmid integrated in the Ech{s} allele) displayed strong in vivo echinocandin resistance . Finally, we were unable to inactivate both alleles at CaFKS1 by two-step integrative disruption, suggesting that CaFks1p is likely to be an essential protein in C . albicans.

Antimicrob Agents Chemother, 1997 Nov, 41(11), 2345 - 51
Heat-induced superaggregation of amphotericin B reduces its in vitro toxicity: a new way to improve its therapeutic index; Gaboriau F et al.; Superaggregation of amphotericin B (AmB) was previously shown to occur upon heating of solutions at 70 degrees C . In the present study, we demonstrate that heat pretreatment of Fungizone (deoxycholate salt of AmB {AmB-DOC}) solutions induces a drastic decrease in the in vitro toxicity of this antibiotic . Heated AmB-DOC colloidal solutions, which mainly contained superaggregated and monomeric forms of the antibiotic, were strongly less hemolytic than unheated solutions (aggregates and monomers) . Thermal pretreatment of AmB-DOC solutions also reduced the toxicity to the cell line HT29, as deduced from two simultaneous cell viability assays (3-4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase release) . These heated colloidal solutions were only slightly less efficient than the unheated ones at inhibiting the growth of Candida albicans cells in vitro . Such results suggest that mild heat treatment of AmB-DOC solutions could provide a new and simple solution for improving the therapeutic index of this antifungal agent by reducing its toxicity to mammalian cells.

Immunology, 1997 Sep, 92(1), 104 - 10
The effects of monoclonal antibodies against iC3b receptors in mice with experimentally induced disseminated candidiasis; Lee KH et al.; CR3 (iC3b receptor), composed of CD11b/CD18, is a beta 2 integrin . A protein that shares antigenic and structural homology with the alpha-chain of CD11b/CD18 has been isolated from the surface of Candida albicans . This molecule is thought to be essential in the pathogenesis of disseminated candidiasis . To evaluate the effects of anti-iC3b receptor antibodies on adhesion between human dermal microvascular endothelial cells (HDMEC) and C . albicans, and in treatment of candidal infection, a binding assay of C . albicans to cultured HDMEC was performed in vitro . An anti-iC3b receptor-specific monoclonal antibody was administered to mice infected with C . albicans . The mice were monitored for mortality and renal involvement by culture and histopathological findings . Flow cytometric analysis demonstrated surface expression of iC3b receptor on C . albicans . The adherence of C . albicans to HDMEC was significantly decreased by treatment with anti-iC3b receptor antibodies . Anti-iC3b receptor antibodies significantly increased the survival time and rate while lowering the renal fungal burden . The iC3b receptors are involved in the adherence of C . albicans to vascular endothelial cells and are likely to be involved in the pathogenesis of disseminated candidiasis . The increased survival in mice infected with C . albicans after treatment with anti-iC3b receptor antibodies indicates that this modality may be beneficial for future development of a new therapy for candidiasis.

FEBS Lett, 1997 Oct 20, 416(2), 203 - 6
Definitive chemical evidence for the constitutive ability of Candida albicans serotype A strains to synthesize beta-1,2 linked oligomannosides containing up to 14 mannose residues; Trinel PA et al.; We have previously reported the presence of phosphate bound beta-1,2 linked oligomannosides with unusually high degrees of polymerization (DP > 7) in the mannan of Candida albicans strain VW32 . To confirm this observation, we have prepared these oligomannosides from the mannan of C . albicans strain NIH A 207 . Gel filtration chromatography and TLC analysis revealed DP up to 14 . For both strains, NMR analysis confirmed the exclusive presence of beta-1,2 linkages in the pools of oligomannosides with a DP higher than 6 which presented an average DP of 10.6 (VW32) and 10.4 (NIH A 207) . These results are important to consider in relation with the ability of these C . albicans derived oligomannosides to trigger TNFalpha synthesis according to their DP.

Cell Mol Life Sci, 1997 Sep, 53(9), 744 - 9
Candida albicans morphogenesis is influenced by estrogen; White S et al.; Conversion of Candida albicans from yeast to mycelial growth is believed to be associated with the organism's virulence . We investigated the role of mammalian hormones in initiating this transformation . Three clinical isolates of Candida albicans were tested for their ability to produce germ tubes under various conditions . Controlled hormonal conditions were provided by stripping rabbit serum with activated charcoal . Steroid compounds under investigation were added back to the stripped serum and yeast were inoculated into the test materials . Microscopic counts of germinated versus ungerminated cells were used as an indicator of morphogenic transformation . The percent of yeast cells germinating was profoundly reduced in stripped compared to unstripped serum . The addition of 1 microM estradiol, cholesterol or testosterone only slightly increased levels of germination above that seen in controls . Estradiol at concentrations 100 times less, however, proved a strong inducer of germination . Cholesterol did not synergize germination when combined with estradiol and the alpha isomer of estradiol had almost no activity as an inducer of morphogenic change in Candida albicans . We conclude that beta estradiol was a morphogenic inducer in three clinical isolates of Candida albicans but only at concentrations typical in vivo.

Mycopathologia, 1997, 137(3), 153 - 7
Evaluation of the AUXACOLOR system for the identification of clinical yeast isolates; Milan EP et al.; In the last decade the number of systemic yeast infections has increased significantly . Although Candida albicans is the most frequently isolated yeast from clinical specimens, the emergence of non-albicans species has clearly been a recent concern . As a consequence, there is a greater need for rapid and accurate methods for yeast identification . The aim of this study was to evaluate the performance of the AUXACOLOR system (Sanofi Diagnostics Pasteur) for the identification of clinically relevant yeasts, as compared with the conventional method . Yeast isolates (n = 97) belonging to 12 species were identified by the commercial system and the classic method . Correct identifications were obtained by using AUXACOLOR system in 79.4% of the isolates tested . Misidentification occurred in 5.2% of the strains and 15.5% were not identified due to a failure in the manufacturer's data base . In order to improve its accuracy, there is a need for expanding the database or revamping the tests included in the system.

J Biol Chem, 1997 Nov 14, 272(46), 28954 - 61
KEX2 influences Candida albicans proteinase secretion and hyphal formation; Newport G et al.; Candida albicans possesses at least seven differentially expressed genes that encode virulence-related secretory aspartyl proteinases (Saps) . Sap DNA sequences predict post-translational processing at lysine-arginine residues in the preproteins, reminiscent of the maturation of Saccharomyces cerevisiae alpha-factor, where a prepropolypeptide is converted into a biologically active pheromone by Kex2, a subtilisin-like proprotein convertase . To investigate involvement of a C . albicans KEX2 homologue in Sap activation, a genetic selection was performed based on KEX2 function . A kex2 strain of S . cerevisiae was transformed with a C . albicans genomic DNA library and screened for the production of active alpha-factor . Positive clones were assayed for killer toxin activity, another Kex2-dependent phenotype . Plasmids that rescued both defects contained a sequence encoding a protein homologous to S . cerevisiae Kex2 . Both alleles of the C . albicans KEX2 were inactivated by successive mutations . Null mutants continued to secrete active Sap2; however, the enzyme was abnormally processed and secreted at reduced levels . Unexpectedly, null mutants were incapable of forming hyphae, instead differentiating into aberrantly shaped cells . The ability to normally process Sap2 and form hyphae was restored upon transformation of null mutants with a KEX2-containing plasmid.

Yeast, 1997 Oct, 13(13), 1199 - 210
Characterization and regulation of the genes encoding ribosomal proteins L39 and S7 of the human pathogen Candida albicans; Delbruck S et al.; Genes encoding the Candida albicans ribosomal proteins L39 and S7 (RPL39, RPS7) were isolated and sequenced . From RPL39 cDNA a single intron interrupting the fifth codon in the genomic sequence could be deduced . Two homologous RPL39 genes in Saccharomyces cerevisiae contain a single intron in a conserved position . In contrast, C . albicans RPS7 was found to lack an intron, while both S . cerevisiae homologs are interrupted by single introns . The deduced L39 and S7 proteins contained 67% and 83% identical residues compared to the S . cerevisiae homologs . During hyphal induction the RPL39, RPS7 and RPL29 transcript levels increased three- to six-fold relative to ribosomal RNA, while ACT1 and RPS33 control transcripts were not regulated extensively . As suggested by unaltered transcript stabilities during hyphal induction, this regulation occurs on the transcriptional level; a conserved 18 bp palindromic sequence (5'-TTAGGGCTATAGCCCTAA-3'), which is present in the promoter regions of the RPL39 and RPS7 genes, may be involved in regulation.

Pharmazie, 1997 Oct, 52(10), 799 - 802
Antiinfective activity of a plant preparation from Geranium sanguineum L; Serkedjieva J; A methanol extract (PC) has been obtained from the Bulgarian medicinal plant Geranium sanguineum L . The antiinfective activity of the plant preparation was studied . The extract inhibited the reproduction of a range of viruses (influenza, herpes simplex, vaccinia, HIV-I) in cell cultures . Its antiinfluenza effect was most pronounced; PC reduced the infectivity of various influenza virus strains in vitro and protected mice in experimental influenza infection . The polyphenol extract inhibited the in vitro growth of Staphylococcus aureus and Candida albicans.

Clin Infect Dis, 1997 Oct, 25(4), 908 - 10
Development of fluconazole resistance in Candida albicans causing disseminated infection in a patient undergoing marrow transplantation; Marr KA et al.; Oral candidiasis due to azole-resistant Candida albicans is an increasing problem in patients with AIDS who received prolonged periods of fluconazole prophylaxis . Infection with C . albicans is also frequent in patients undergoing transplantation . However, azole resistance has not been appreciated as a major problem for these patients, presumably because they receive a relatively short duration of fluconazole prophylaxis . We describe a case of disseminated candidiasis due to fluconazole-resistant C . albicans in a patient following marrow transplantation . Restriction fragment length polymorphism analysis with use of the C . albicans strain-specific Ca3 probe was performed on sequential isolates . Identical banding patterns were obtained, thereby confirming that a fluconazole-susceptible endogenous C . albicans acquired azole resistance during a brief exposure to the drug and subsequently caused disseminated infection . This observation raises questions regarding the incidence, significance, and mechanism of azole resistance in fungi causing infection in this population.

Infect Immun, 1997 Nov, 65(11), 4468 - 75
Misexpression of the white-phase-specific gene WH11 in the opaque phase of Candida albicans affects switching and virulence; Kvaal CA et al.; Candida albicans WO-1 switches between a white- and an opaque-colony-forming phenotype . The gene WH11 is expressed differentially in the white phase . The WH11 open reading frame was inserted downstream of the promoter of the opaque-phase-specific gene OP4 in the transforming vector pCWOP16, and resulting transformants were demonstrated to misexpress WH11 in the opaque phase . Misexpression had no effect on the ability to switch from the white to the opaque or the opaque to the white phase, and it had no effect on the genesis of the unique opaque-phase cellular phenotype, even though the Wh11 protein was distributed throughout the cytoplasm in a manner similar to that observed for the endogenous gene product in the white phase . Misexpression did, however, increase the frequency of the opaque-to-white transition 330-fold and markedly increased the virulence of cells in the opaque phase in a mouse tail injection model.

Lett Appl Microbiol, 1997 Sep, 25(3), 181 - 5
Alveolar macrophage reaction to Candida species; Nessa K et al.; Candida species are increasingly important fungal pathogens . The reaction of rat alveolar macrophages (AM) to Candida albicans was compared with that to C . glabrata and C . krusei . Phagocytosis of C . glabrata was similar to that of C . albicans, but significantly slower for C . krusei due to reduced attachment . After opsonization, attachment of C . albicans and C . krusei to AM was significantly increased and there was no significant difference between the two species . The oxidative metabolism of AM with candida species was two to three times higher than that of the resting AM both during and 24 h after the phagocytosis . All three species showed a considerable fraction (5-10%) of phagolysosomes with pH > or = 6.5 after 3 h and a smaller percentage (1%) after 24 h.

J Clin Microbiol, 1997 Nov, 35(11), 2943 - 8
Detection and identification of Candida species in experimentally infected tissue and human blood by rRNA-specific fluorescent in situ hybridization; Lischewski A et al.; Two 18S rRNA-targeted oligonucleotide probes specific for Candida albicans and Candida parapsilosis were used to detect and identify by fluorescent in situ hybridization these medically important Candida species in deep organs of mice after experimental systemic infection . The C . albicans-specific probe detected fungal cells in kidney, spleen, and brain sections of a mouse infected with C . albicans but not in a mouse infected with the closely related species C . parapsilosis . Conversely, the C . parapsilosis-specific probe detected fungal cells in the deep organs of a mouse infected with C . parapsilosis but not in the deep organs of a C . albicans-infected mouse . In addition, the C . albicans-specific probe was used to detect this species in human blood spiked with yeast cells by a lysis-filtration assay and subsequent fluorescent in situ hybridization . By this assay, as few as three yeast cells per 0.5 ml of blood were consistently detected . Our results demonstrate that fluorescent in situ hybridization with species-specific rRNA-targeted oligonucleotide probes provides a novel, culture-independent method for the sensitive detection and identification of Candida species in clinically relevant material.

J Clin Periodontol, 1997 Oct, 24(10), 753 - 60
Effects of tetracycline hydrochloride and chlorhexidine gluconate on Candida albicans . An in vitro study; MacNeill S et al.; This study examined the effects of tetracycline hydrochloride (TCN) and chlorhexidine gluconate (CHX) on the growth and viability of Candida albicans . Subcultures of Candida albicans on Sabouraud's agar, were divided into 5 treatment groups: group 1, untreated control; group 2, 0.12% CHX; group 3, 3.0 mg/ml TCN adjusted to pH 4.5; groups 4 and 5, sodium azide free Tris buffer adjusted to pH 4.5 and pH 7.4, respectively . All groups were incubated for 10 days, and sampled and subcultured daily to determine the viability of each group . Additional samples from group 2 (day 4), group 4 (day 7) and all groups at day 10 were selected for SEM and TEM examination . Visual, SEM and TEM results showed that for groups 1, 3, 4, and 5 there was a heavy and constant uniform growth of Candida albicans throughout the period of the study . However, group 2 (CHX), showed decreasing viability and attachment from day 3 to day 10, with SEM and TEM revealing decreased blastospores and profound changes in the ultrastructural morphology, indicating inhibition of normal cell growth and replication . These results show that TCN even when used at high concentrations, in vitro, will allow uninhibited growth of Candida albicans whereas CHX inhibits cell growth and replication.

Diagn Microbiol Infect Dis, 1997 Sep, 29(1), 5 - 9
Candidemia in a Canadian tertiary care hospital from 1976 to 1996; Karlowsky JA et al.; The incidence of candidemia was reviewed at the Health Sciences Centre in Winnipeg, Canada, over a 21-year period (1976 to 1996) . Candida species were identified as blood-stream isolates in significantly (p < 0.05) higher numbers from 1991 to 1996 than in the previous 15 years . Antifungal susceptibilities remained unchanged with Candida albicans isolates tested from 1985 to 1996 . Retrospective chart reviews revealed that all patients with candidemia possessed at least two risk factors . The main risk factors identified were recent or concurrent antibiotic therapy (95% of patients), presence of a central line (93% of patients), and immunosuppression (88% of patients) . Treatment generally involved amphotericin B therapy (81% of patients), and death occurred in 52% of the patients . Mortality directly attributable to Candida species could be established in 23% of patients.

Intensive Care Med, 1997 Sep, 23(9), 1002 - 4
Surgical management of Candida suppurative thrombophlebitis of superior vena cava after central venous catheterization; Garcia E et al.; Septic deep venous thrombosis is a major complication associated with central venous catheterization in intensive care units . The most common causative organisms are Staphylococcus aureus, gram-negative bacilli and Candida species . The incidence of Candida infections is increasing, especially in intensive care patients receiving total parenteral nutrition and long-term broad-spectrum antibiotics . Although intravascular catheter-induced septic thrombophlebitis is quite common, superior vena cava obstruction is a rare complication . However, few data exist concerning the best strategy for managing septic thrombophlebitis, especially when medical therapy fails . We report successful surgical management of Candida albicans suppurative thrombosis of the superior vena cava in a young patient.

Eur J Clin Microbiol Infect Dis, 1997 Aug, 16(8), 601 - 5
Genotypic identification of sequential Candida albicans isolates from AIDS patients by polymerase chain reaction techniques; Barchiesi F et al.; Random amplification of polymorphic DNA and inter-repeat polymerase chain reaction (IR-PCR) were compared with restriction fragment length polymorphism (RFLP) analysis as methods for DNA typing of Candida albicans . Forty-seven strains of Candida albicans isolated from the oral cavities of five AIDS patients undergoing fluconazole therapy were analyzed . There was an excellent correspondence between the DNA types obtained by both PCR-based techniques and by RFLP . With the exception of one patient who was infected with three DNA types of Candida albicans during a five-year observation period, the patients each harboured only one major strain, which became progressively less susceptible to fluconazole . Each DNA type was unique to a patient . The data suggest that these typing methods are suitable for investigating the epidemiology of oropharyngeal candidiasis in this patient group.

Eur J Clin Microbiol Infect Dis, 1997 Aug, 16(8), 598 - 600
Mixed Candida glabrata and Candida albicans disseminated candidiasis in a heroin addict; Bougnoux ME et al.; The case of a white-heroin addict who developed disseminated candidiasis following coinfection by Candida glabrata and Candida albicans is reported . Genomic random amplified polymorphic DNA typing suggested that the Candida glabrata blood isolates originated in the oral cavity of the patient . This case strengthens the evidence that Candida species other than Candida albicans can be involved in the pathogenesis of disseminated candidiasis in heroin addicts.

Chem Immunol, 1997, 68, 110 - 35
Initiation of T-helper cell immunity to Candida albicans by IL-12: the role of neutrophils; Romani L et al.; The Th1/Th2 paradigm of acquired immunity is proving essential for a better understanding of immunoregulation in candidiasis and perhaps other fungal infections, conditions that may be life-threatening in humans and difficult to control by chemotherapy alone, especially in neutropenic and severely immunocompromised patients . In its basic conception applied to Candida infection in mice, this paradigm calls for: (i) an association between Th1 responses and the onset/maintenance of phagocyte-dependent immunity, critical for opposing infectivity of the commensal, focusing an infection, and clearing the yeast from infected tissue; (ii) the ability of the yeast to activate Th2 response as an evasive strategy; (iii) the reciprocal regulation of Th1 and Th2 responses, resulting in a dynamic balance between these two types of reactivity . This balance, in concert with a variety of environmental factors, may regulate the status of the yeast as a commensal or pathogen in the mucosal tissue of colonized humans, but may also determine the outcome of deep-seated systemic infections once hematogenous dissemination of the yeast has occurred . An important corollary of this hypothesis may be the possible combined effects on Th immunity of Candida carriage/infection and various disease states . While immune deficiency or dysregulation, resulting in an altered cytokine balance as may occur in AIDS, can reasonably be expected to increase local infectivity of the yeast, it is even more intriguing that antifungal chemotherapy will resolve some of the unusual skin (atopic dermatitis-like) disorders frequently observed in this clinical setting, patients with AIDS . Besides, an immunopathologic role for Candida has been suggested for atopic dermatitis, atopy, and other conditions, overtly associated or not with Candida . Thus, the Th cell dichotomy to Candida may have important implications not only for regulation of the balance between commensalism and infection, but may also contribute to the onset or dominance of Th2 responses in other disease states . A similar example, although with different effects, may be provided by the temporary improvement seen in atopic dermatitis patients in concomitance with acute severe infections, an effect that has been proposed to result from transient down-regulation of the predominant Th2 cell reactivity . With a view to either controlling Candida infections or opposing Candida-related immunopathology, the promotion of yeast-specific Th1 responses appears to be a promising immunotherapeutic approach . This, in principle, could be achieved by subtraction of Th2 cytokines or by administration of Th1-promoting cytokines . However, our initial studies with exogenous IL-12 were unsuccessful, suggesting that the recombinant cytokine: (i) is unable to oppose Th2 differentiation driven in vivo by IL-4/IL-10; (ii) may induce endogenous IL-10 production as a regulatory response, and (iii) may potentate local inflammatory responses in gastrointestinal infection or even trigger IFN-gamma-dependent mechanisms of fungal septic shock. . More recent studies seem to provide encouraging results, at least under specific conditions of testing . In acute candidemia, neutrophils appear to be a major source of the directive cytokines, IL-12 and IL-10, thus contributing to the selection of Th1 and Th2 cell responses to LVS or virulent infection, respectively . Neutrophils may also be an important source of IL-10 released in response to challenge with exogenous IL-12 . As a result, the Th1-promoting role of IL-12 may be largely unopposed (by IL-10 induction) in neutropenic mice, which would otherwise succumb to LVS challenge . These animals are, in fact, cured by replacement therapy with IL-12 and acquire durable, Th1-associated anticandidal protection . These findings may be very important for immunotherapy of fungal infections in humans . Neutropenic patients are those at the highest risk for developing systemic candidal infections . (ABSTRACT TRUNCATED)

J Otolaryngol, 1997 Oct, 26(5), 300 - 5
Prevalence, density, and manifestations of oral Candida albicans in patients with Sjögren's syndrome; Rhodus NL et al.; OBJECTIVE: Various investigators have reported a high prevalence of oral Candida species in patients with salivary gland dysfunction (SGD) . The purpose of this study was to assess the prevalence of oral Candida albicans, its oral manifestations, and to compare the number of colony-forming units of Candida albicans in patients with primary Sjogren's syndrome and secondary Sjogren's syndrome with the whole unstimulated salivary flow rate in each group . METHOD: An age-sex-matched group of control subjects was selected for comparison . Oropharyngeal collection of samples and culturing was performed on each subject . Quantitative cultures specific for Candida albicans were performed . RESULTS: The frequency distribution indicated that > 80% of all SS subjects were positive for Candida albicans vs . none of the controls . The most common lesion was angular cheilitis followed by chronic atrophic candidiasis . The subjects with Sjogren's syndrome also demonstrated significantly high numbers of Candida albicans colony-forming units . CONCLUSIONS: This study indicates significantly higher Candida albicans colonization in patients with primary or secondary Sjogren's syndrome as compared to a control population . Candida albicans colonization was higher in patients with secondary Sjogren's syndrome than in patients with primary Sjogren's syndrome; however, the amount of Candida albicans was not universally relative to salivary flow.

Clin Exp Obstet Gynecol, 1997, 24(2), 112 - 3
Endometrium is not a reservoir for recurrent vaginal candidiasis; Tasdemir M et al.; The aim of this study was to determine whether the endometrium acts as a reservoir for Candida albicans in cases of recurrent vaginal candidiasis . Twenty-five women with documented history of recurrent vaginal candidiasis were enrolled in the study and endometrial samples were cultured for Candida albicans . Only two patients had positive cultures for Candida albicans . Therefore, we concluded that the endometrium is not a common reservoir for Candida albicans.

Arzneimittelforschung, 1997 Sep, 47(9), 1056 - 60
Studies on the antifungal activity of the new imidazole antimycotic lanoconazole in infected sites . Distribution in the skin and in vitro activity in the presence of stratum corneum; Niwano Y et al.; To answer the question why topically applied lanoconazole (CAS 101530-10-3, NND-318) is so highly effective in the treatment of dermatomycoses in both animal models and human patients, the antifungal activity of lanoconazole in infected sites was investigated . 1 . Distribution of lanoconazole in rat skin: The distribution of lanoconazole within the dorsal skin of rats was examined by measurement of radioactivity and microscopic autoradiography . 24 h after dermal application of 14C-lanoconazole cream formulation, 83% of the total radioactivity in the skin was recovered from the stratum corneum, and thereafter the radioactivity decreased gradually up to 96 h . Metabolite analysis showed that more than 94% of the extractable radioactivity was lanoconazole itself after 24 and 48 h . Microautoradiograms of the skin also supported the radioactivity distribution . 2 . In vitro antifungal activity in stratum corneum-containing medium: Candida albicans TIMM 2640 was incubated for 10 days at 27 degrees C in a vitamin-supplemented yeast carbon base medium containing 5 mg/ml of human stratum corneum and different concentrations of lanoconazole or bifonazole (CAS 60628-96-8, reference agent) . Compared with the control culture, lanoconazole strongly inhibited fungal growth in a concentration dependent manner at concentration above 0.1 microgram/ml, resulting in a reduction of viable cell recovery to less than 50% after 10 days . The inhibitory activity of bifonazole was clearly weaker than that of lanoconazole . At concentrations of 0.1-10 micrograms/ml lanoconazole reduced keratinolytic acid proteinase activity in the culture-supernate to 40-70% of the control value, while bifonazole showed 50% reduction of the activity at a concentration of 10 micrograms/ml . These results indicate that lanoconazole is mainly distributed and retained in the stratum corneum after topical application where it exerts strong antifungal activity.

J Med Chem, 1997 Oct 10, 40(21), 3456 - 65
Syntheses and biological activities (topoisomerase inhibition and antitumor and antimicrobial properties) of rebeccamycin analogues bearing modified sugar moieties and substituted on the imide nitrogen with a methyl group; Anizon F et al.; As a part of studies on structure-activity relationships, several potential topoisomerase I inhibitors were prepared . Different analogues of the antitumor antibiotic rebeccamycin substituted on the imide nitrogen with a methyl group were synthesized . These compounds bore either the sugar residue of rebeccamycin, with or without the chlorine atoms on the indole moieties, or modified sugar residues (galactopyranosyl, glucopyranosyl, or fucopyranosyl) linked to the aglycone via a beta- or alpha-N-glycosidic bond . Their inhibitory properties toward protein kinase C, topoisomerase I, and topoisomerase II were examined, and their DNA-binding properties were investigated . Their in vitro antitumor activities against murine B16 melanoma and P388 leukemia cells were determined . Their antimicrobial activities were tested against Gram-positive bacteria Bacillus cereus and Streptomyces chartreusis, Gram-negative bacterium Escherichia coli, and yeast Candida albicans . These compounds are inactive toward topoisomerase II but inhibit topoisomerase I . A substitution with a methyl group on the imide nitrogen led to a loss of proteine kinase C inhibition in the maleimide indolocarbazole series but did not prevent topoisomerase I inhibition . Compounds possessing a beta-N-glycosidic bond, which fully intercalated into DNA, were more efficient at inhibiting topoisomerase I than their analogues with an alpha-N-glycosidic bond; however, both were equally toxic toward P388 leukemia cells . Dechlorinated rebeccamycin possessing a methyl group on the imide nitrogen was about 10 times more efficient in terms of cytotoxicity and inhibition of topoisomerase I than the natural metabolite.

J Med Chem, 1997 Oct 10, 40(21), 3434 - 41
Cephalosporin 3'-phloroglucide esters and 7-(phloroglucidamido) cephalosporins as novel antibacterial agents; Hwu JR et al.; Two series of new phloroglucide derivatives were synthesized that possessed antibacterial activities . The first series includes cephalosporin 3'-phloroglucide esters 19 and 20, which were obtained by condensation of cephalosporin 16 with bioactive phloroglucides 14 and 15, respectively . They exhibited a dual mode of antibacterial action . In comparison with cephalosporins 26 and 27, bearing an acetoxy unit at the C-3' position, the bifunctional cephalosporins 19 and 20 showed a broadened spectrum of activity . Results from the consistent valence force field (CVFF) calculations indicate that the most stable conformational isomer of phenolic acid 14, holding a cis-syn-syn geometry, possessed a cavity . It provides an ideal environment to accommodate metal ions of holoenzymes . Phenolic keto acid 15, however, possessed a trans-anti-syn conformation, which allowed chelation between metal ions and the phenolic hydroxyl groups as well as the carbonyl functionalities . Our biological results show that the cavity formed in phloroglucides plays an important role . The second series includes 7-(phloroglucidamido)cephalosporins 24 and 25, which were synthesized by condensation of cephalosporin 21 with 14 and 15, respectively . Results from the CVFF calculations indicate that cephalosporin 24 also possessed a cavity . Unlike cephalosporin 3'-phloroglucide esters 19 and 20, cephalosporins 24 and 25 were found resistant to beta-lactamases from Staphylococcus aureus 95 and Pseudomonas aeruginosa 18S-H . These new compounds, however, showed notable activities against S . aureus FDA 209P, S . aureus 95, Candida albicans, P . aeruginosa 1101-75, and P . aeruginosa 18S-H.

Pneumonol Alergol Pol, 1997, 65(5-6), 355 - 9
{In vitro susceptibility to antifungal agents of Candida strains isolated from patients with various diseases of the respiratory tract}; Batura-Gabryel H et al.; The aim of the study was the estimation of in vitro susceptibility to antifungal agents of yeast isolated from sputum of 70 respiratory diseases patients using the disc-diffusion method-antimycogram . The following agents were tested: amphotericin B, 5-fluorocytosine, nystatin, ketoconazole, fluconazole . Only Candida strains were isolated from sputum, 82% of them were Candida albicans . We noted differences in susceptibility to antimycotics of Candida strains . The best antimycotic in vitro was 5-fluorocytosine . 54% of isolated Candida strains were resistant to 1 or more antimycotics.

Pediatr Cardiol, 1997 Nov-Dec, 18(6), 440 - 2
Candida endocarditis in an infant; Hartyanszky IL et al.; Infective endocarditis is a rare disease in infants . A 1-year-old boy with a large Candida albicans vegetation in the right atrium and superior vena cava was operated on successfully . During the newborn period he had had a right transverse colostomy for Hirschsprung's disease . Ten months later a subsequent rectosigmoidectomy and direct anastomosis were performed, but because of peritonitis that followed a leak at the site of the anastomosis parenteral nutrition was needed for 8 weeks . The probable source of Candida was an infected intravenous line.

Mycopathologia, 1997, 137(2), 95 - 105
Ultrastructural localization of the secretory aspartyl proteinase in Candida albicans cell wall in vitro and in experimentally infected rat vagina; Stringaro A et al.; Detection and ultrastructural localization of aspartyl proteinase (Sap) in Candida albicans experimentally infecting rat vagina were studied . Two Sap-positive (Sap+) and one Sap-negative (Sap-) strains of the fungus, endowed with high and low experimental vaginopathic potential, respectively, were used . Both Sap+ strains produced consistent Sap levels in the rat vagina, while the Sap- strain did not produce any measurable Sap . Electron microscopy of thin sections of chemically-fixed vaginal scrapings showed clear evidence of hyphae of proteolitic strains of C . albicans invading the keratinized epithelial cell layer of the vagina . The fungal cells exhibited a pronounced fibrillar layer on the cell wall with a marked intermixing of fungal and vaginal materials especially pronounced at the hyphal tip . Post-embedding immunogold techniques with the use of anti-Sap polyclonal and the specifically generated monoclonal antibody GF1 showed that Sap was essentially localized in the cell wall of C . albicans early during infection, in a cytological pattern mirroring Sap localization in C . albicans cells grown in Sap-inductive media in vitro . In summary, the data offer a new biochemical and ultrastructural evidence that Sap is actively secreted during experimental rat vaginitis by C . albicans . Cell wall localization of Sap is probably inherent to this active secretion process.

Mycopathologia, 1997, 137(2), 87 - 94
Phenotype and genotype of Candida albicans strains isolated from pregnant women with recurrent vaginitis; Maffei CM et al.; Fourteen out of 80 pregnant women receiving prenatal care presented signs and symptoms of recurrent vaginal candidiasis . Candida albicans strains were isolated from 12 patients (85.7%), and these were submitted to morphotyping (morphological characteristics of the colony), antifungal typing (pattern of sensitivity to amphotericin B, 5-fluorcytosine, myconazole, ketoconazole and fluconazole) and genotyping (electrophoretic migration of DNA fragments digested with EcoRI and HinfI) . Alteration of morphotype and antifungal type was observed in 50% of the patients, but the genotype of the strains isolated from the same patients at different times was identical in all subjects . The predominant morphotypes presented continuous fringes and the basic changes observed among antifungal types was the emergence of strains resistant to myconazole, which was the drug used for the treatment of the first episode of vaginitis . We conclude that recurrent vaginal candidasis is caused by the persistence of a single yeast genotype that undergoes morphological and behavioral changes in the presence of antifungal agents due to the selective pressure to which it is submitted.

Arq Bras Cardiol, 1997 Jan, 68(1), 35 - 7
{Superior vena cava and right atrium thrombosis successfully treated with streptokinase}; Baruzzi AC et al.; The case of a 56 year-old male with acute lymphoid leukemia and no signs of activity for the last four months is reported . He presented with superior vena cava thrombosis caused by a Hickman catheter, and had positive blood cultures for Candida albicans and Staphylococcus epidermidis . Despite adequate antimicrobial therapy, the fever persisted, and the patient was submitted to surgical thrombectomy . One week following the procedure, the fever returned, and thrombosis of the superior vena cava extending to the right atrium was identified by transesophageal echocardiography (TEE) . The patient underwent thrombolytic therapy with streptokinase, and no thrombus could be identified in the control TEE . No hemorrhagic or thromboembolic complication occurred . The patient was discharged with oral anticoagulation.

Hautarzt, 1997 Jun, 48(6), 397 - 401
{Effect of domestic laundry processes on mycotic contamination of textiles}; Ossowski B et al.; Inadequately decontaminated clothing may be a source of reinfection following therapy of dermato- and onychomycoses . The objective of this study was to determine whether domestic laundering is suitable for cleansing mycotically contaminated garments . Textile-samples contaminated with Trichophyton rubrum, Trichophyton mentagrophytes, Candida albicans and Scopulariopsis brevicaulis were washed in an ordinary washing machine at different temperatures . Regardless of the textiles and detergents used, reliable decontamination was achieved by laundering at 60 degrees C . Trichophyton rubrum was eliminated with a washing temperature of 30 degrees C.

Antimicrob Agents Chemother, 1997 Oct, 41(10), 2224 - 8
Studies of the mechanism of human salivary histatin-5 candidacidal activity with histatin-5 variants and azole-sensitive and -resistant Candida species; Tsai H et al.; Histatins are a group of small, cationic, antifungal peptides present in human saliva . A previous molecular modeling analysis suggested structural similarity between the Phe14-His15 and His18-His19 dipeptide sequences in histatin-5 (Hsn-5; a 24-amino-acid polypeptide) and the sequence of miconazole (one of the azole-based antifungal therapeutic agents), implying that the mechanisms of killing of Candida albicans by these two molecules may be similar . To further elaborate on this observation, we have produced two variants of Hsn-5 in which Phe14-His15 or His18-His19 dipeptide sequences were replaced by Ala-Ala (F14A/H15A and H18A/H19A) to eliminate the phenyl and imidazole rings of the side chains and assessed their candidacidal activities against C . albicans . In addition, we tested azole-resistant C . albicans and Candida glabrata strains for their susceptibilities to Hsn-5 . Analysis of the purified recombinant proteins for their candidacidal activities indicated that both variants were significantly less effective (the molar concentrations required to kill half of the maximum number of cells {ED50s}, approximately 67 and approximately 149 microM for F14A/H15A and H18A/H19A, respectively) than the unaltered Hsn-5 (ED50, approximately 8 microM) at killing C . albicans, suggesting that the two dipeptide sequences are important for the candidacidal activity of Hsn-5 . Assessment of the candidacidal activity of Hsn-5 with the well-characterized azole-resistant strains of C . albicans and C . glabrata, however, suggested that the mode of action of histatins against Candida is distinct from that of azole-based antifungal agents because Hsn-5 kills both azole-sensitive and azole-resistant strains equally well.

Support Care Cancer, 1997 Sep, 5(5), 381 - 6
Immuno-nutrition: designer diets in cancer; Imoberdorf R; Weight loss, associated with advanced stage of neoplastic disease, is negatively correlated with survival in cancer patients . Alterations in substrate metabolism contribute to the impaired nutritional status . Energy expenditure, assessed by indirect calorimetry, seems to be very variable . Hypermetabolism may occur frequently, but the increase in energy expenditure rarely exceeds 10-15% . Another hallmark of cancer is depression of both cellular and humoral immune functions . Glutamine, the most abundant amino acid in the body, is an important substrate for rapidly proliferating cells and tissues . Arginine has been shown to stimulate the immune system, to enhance wound healing and to decrease the rate of tumour growth . Dietary supplementation with omega-3 fatty acids shifts the production of prostaglandins from the dienoic to the trienoic variety, which is much less immunosuppressive . Finally, administration of oligonucleotides improved survival to a challenge with Candida albicans . The needs for nutritional support in cancer patients should be considered from at least two perspectives: curative versus palliative treatment . Several prospective, randomized, double-blind studies in cancer patients undergoing major upper gastrointestinal surgery demonstrated significant improvements in postoperative immunological responses, a reduction in the frequency of infections and wound complications and in the length of hospital stay in the group receiving a diet enriched with arginine, RNA and omega-3 fatty acids . However, the impact on mortality remains to be established . In palliative situations, no clinical data documenting beneficial effects of long-term nutritional support with designer diets have been published, probably as a consequence of persisting concern about promoting tumour growth . Moreover, because of the heterogeneity of this patient population such studies are difficult to perform . Finally, nutritional intervention for patients with terminal cancer remains highly controversial . The basis for improvements in nutritional support is a better understanding of the metabolism of cancer patients, especially in patients with advanced disease.

Infect Immun, 1997 Oct, 65(10), 4360 - 4
Hemin induces germ tube formation in Candida albicans; Casanova M et al.; Hemin induced germination of Candida albicans blastoconidia when cells grown up to the early exponential phase were shifted from 28 to 37 degrees C (70 to 75% of cells exhibited germ tubes) . N-Acetyl-D-glucosamine (GlcNAc), another inducer of myceliation in this fungus, caused a similar effect . The combination of hemin and GlcNAc resulted in a higher percentage (95%) of blastoconidial germination . These results suggest that in addition to temperature, hemin levels and carbon source may coordinately regulate the expression of subsets of genes involved in the yeast-to-mycelium transition in C . albicans.

Infect Immun, 1997 Oct, 65(10), 4100 - 7
Biochemical characterization of Candida albicans epitopes that can elicit protective and nonprotective antibodies; Han Y et al.; We previously reported that the immunoglobulin M (IgM) monoclonal antibody (MAb) B6.1 protects mice against disseminated candidiasis, whereas the IgM MAb B6 does not . Both MAbs are specific for an adhesin fraction isolated from the cell surface of Candida albicans, but their epitope specificities differ . In the present study, we examined the surface locations of both epitopes and obtained structural information regarding the B6.1 epitope . Immunofluorescence confocal microscopic analysis of C . albicans yeast forms showed that epitope B6.1 is displayed rather homogeneously over the entire cell surface, whereas epitope B6 appears to have a patchy distribution . Both antibodies were essentially nonreactive with the surfaces of mycelial forms of the fungus, indicating that neither epitope is expressed on the surfaces of these forms . For isolation of the B6.1 epitope, the adhesin fraction consisting of cell surface phosphomannan was subjected to mildly acidic (10 mM HCl) hydrolysis and was fractionated into acid-labile and acid-stable portions by size exclusion chromatography . Antibody blocking experiments showed that the B6.1 epitope is an acid-labile moiety of the phosphomannan and that the B6 epitope is located in the acid-stable fraction . The B6 epitope appeared to be mannan because it was stable to heat (boiling) and protease treatments but was destroyed by alpha-mannosidase digestion . The B6.1 epitope eluted from the size exclusion column in two fractions . Mass spectroscopic analyses showed that one fraction contained material with the size of a mannotriose and that the other was a mixture of mannotriose- and mannotetraose-size substances . Dose response inhibition tests of the fractions indicated that the B6.1 epitope is associated with the mannotriose . Nuclear magnetic resonance (NMR) spectroscopic analysis of the epitope yielded data consistent with a beta-(1-->2)-linked mannotriose . The fine structure of the B6 epitope is under investigation . Information derived from these investigations will be useful both in understanding protective versus nonprotective antibody responses to C . albicans and in improving anti-Candida vaccine formulations.

Mol Cell Biol, 1997 Oct, 17(10), 5960 - 7
PHR2 of Candida albicans encodes a functional homolog of the pH-regulated gene PHR1 with an inverted pattern of pH-dependent expression; Muhlschlegel FA et al.; Deletion of PHR1, a pH-regulated gene of Candida albicans, results in pH-conditional defects in growth, morphogenesis, and virulence evident at neutral to alkaline pH but absent at acidic pH . Consequently, we searched for a functional homolog of PHR1 active at low pH . This resulted in the isolation of a second pH-regulated gene, designated PHR2 . The expression of PHR2 was inversely related to that of PHR1, being repressed at pH values above 6 and progressively induced at more acidic pH values . The predicted amino acid sequence of the PHR2 protein, Phr2p, was 54% identical to that of Phr1p . A PHR2 null mutant exhibited pH-conditional defects in growth and morphogenesis analogous to those of PHR1 mutants but manifest at acid rather than alkaline pH values . Engineered expression of PHR1 at acid pH in a PHR2 mutant strain and PHR2 at alkaline pH in a PHR1 mutant strain complemented the defects in the opposing mutant . Deletion of both PHR1 and PHR2 resulted in a strain with pH-independent, constitutive growth and morphological defects . These results indicate that PHR1 and PHR2 represent a novel pH-balanced system of functional homologs required for C . albicans to adapt to environments of diverse pH.

Indian J Exp Biol, 1997 Apr, 35(4), 361 - 5
Antifungal activity of conjugated styryl ketones; Manavathu EK et al.; The susceptibility of Aspergillus fumigatus to a series of alpha, beta-unsaturated styryl ketones known to be thiol-alkylators was examined, and the results were compared with those obtained for Candida albicans . Among 13 compounds used in our study, one (designated NC1110) inhibited the growth of A . fumigatus completely at low concentrations (minimum inhibitory concentration = 32 microM) . Structure-activity analysis of these compounds indicated that the electron attracting property as well as the overall hydrophobicity of the compounds are important parameters for their antifungal activity . These preliminary results suggest that further modification of these molecules to enhance their hydrophobicity and the electron attracting property may result in more active compounds with improved antifungal activity.

J Antibiot (Tokyo), 1997 Aug, 50(8), 665 - 70
Isolation and structure elucidation of 7,8-dideoxy-6-oxo-griseorhodin C produced by Actinoplanes ianthinogenes; Panzone G et al.; A novel product, isolated from a culture broth of Actinoplanes ianthinogenes fermented for producing purpuromycin, was purified and its structure established on the basis of physico-chemical data and chemical reactions . The new product resulted to be structurally related to griseorhodins, a group of hydroquinonic antibiotics obtained from Streptomyces californicus . This compound showed a weak activity against Gram-positive and resulted inactive against Gram-negative bacteria and Candida albicans.

Ophthalmologe, 1997 Jun, 94(6), 397 - 400
{Endogenous endophthalmitis in severe generalized diseases}; Augsten R et al.; BACKGROUND: In spite of modern antibacterial, antimycotic and operative therapies, endogenous endophthalmitis connected with other severe diseases and an impaired state of health is an eye complication with a very poor prognosis . PATIENTS: The study presents observations about the progress of the endogenous endophthalmitis in five eyes of four patients . The age of the patients ranged from 58 to 71 years . The span between the appearance of the first eye symptoms and hospital treatment amounted to 1 week and 3 months . The general diseases have been diagnosed as endometrium cancer, colon cancer, corpus uteri cancer (with intestinal fistula) and pancreatic insufficiency (with subclavian catheter) as well as diabetes mellitus . RESULTS: In regard to the infections we analyzed (Candida albicans in 3 eyes, bacteria in 2 eyes), very intensive medical treatment was initiated, including antibacterial or antimycotic therapy (local, subconjunctival, intraocular and systemic) as well as pars plana vitrectomy and/or vitreous puncture . Nevertheless, by the end of the clinical attendance, the following statements were established: one-eye enucleation, one-eye evisceration, two eyes perception of light, one eye visual acuity 0.2 . CONCLUSION: The patients should be immediately admitted to a hospital with a department for vitreous surgery . Only with this option is it possible to initiate adequate therapy.

Chemotherapy, 1997 Sep-Oct, 43(5), 346 - 51
Effects of antifungal pretreatment on the susceptibility of Candida albicans to human leukocytes; Minguez Minguez F et al.; The aim of this work was to measure the susceptibility of Candida albicans pretreated for 2 or 6 h with fluconazole (Flu), ketoconazole (Ktz), amphotericin B (AmB) and 5-fluorocytosine (5-Fc) to the fungicidal action of human polymorphic leukocytes . The influence of pretreatment was measured by comparing the delay (in hours) and reduction (log10) in growth of pretreated cultures in the presence of leukocytes and serum with that of non-pretreated control cultures . Six-hour pretreatments with Flu, Ktz, AmB and 5-Fc led to growth delays of 1, 3, 3 and 3 h, respectively . No significant differences were found when pretreatment lasted only 2 h . With respect to a reduction in growth, this was larger when the preincubation was 6 h, mainly for 5-Fc and AmB . 5-Fc was seen to be the most effective, followed by Ktz, AmB and finally Flu . It may be concluded that antifungal pretreatment renders this yeast more susceptible to the action of leukocytes . The degree of susceptibility achieved is dependent on the antifungal agent employed.

Microbiology, 1997 Sep, 143 ( Pt 9), 3023 - 32
Phenotypic characterization of a Candida albicans strain deficient in its major exoglucanase; Gonzalez MM et al.; Both alleles of the XOG1 gene of Candida albicans, which encodes a protein with exoglucanase activity, were sequentially disrupted . Enzymic analysis of either cell extracts or culture supernatants of disrupted strains revealed that this gene is responsible for the major exoglucanase activity in C . albicans, although residual exoglucanase activity could still be detected . xog1 null mutants showed similar growth rates in both rich and minimal liquid medium as compared to the wild-type strain, indicating that the enzyme is not essential for C . albicans growth . In addition, no differences were observed between wild-type and xog1 null mutants with respect to their ability to undergo dimorphic transition . However, small but repeatable differences were found between the wild-type and the null mutant with respect to susceptibility to chitin and glucan synthesis inhibitors . Using a murine model of experimental infection, no significant differences in virulence were observed . The xog1 null strain is thus a suitable recipient for studying Candida gene expression using the exoglucanase as a reporter gene.

Microbiology, 1997 Sep, 143 ( Pt 9), 3015 - 21
Cell wall protein mannosylation determines Candida albicans cell surface hydrophobicity; Masuoka J et al.; Cell surface hydrophobicity (CSH) has been shown to be an important factor in the ability of the opportunistic pathogenic yeast Candida albicans to adhere to surfaces . Hydrophobic cells adhere more readily to host tissue, and are more resistant to phagocytic killing, than hydrophilic cells . Consequently, CSH plays an important role in the pathogenicity of C . albicans . Previous work suggested a relationship between CSH and cell wall protein glycosylation . The present work tests the hypothesis that changes in outer chain mannosylation, rather than complete loss of oligosaccharide groups, are sufficient to modulate CSH . These studies compared wild-type cells to a variant that has altered mannosylation and is hydrophobic under conditions in which wild-type cells are hydrophilic . Composition analysis of cell surface digests showed that the glycosylation of wild-type cell surface proteins was much more extensive than that seen in the variant . Antibodies which recognize the acid-labile and acid-stable portions of C . albicans mannan showed not only differences between wild-type and variant cells but also differences between wild-type hydrophilic and wild-type hydrophobic cells . The results suggest that exposure of surface hydrophobic regions on C . albicans may be related to the abundance of phosphodiester-linked, acid-labile mannosyl groups rather than the complete loss of outer chain mannosylation on cell wall proteins.

Microb Drug Resist, 1997 Fall, 3(3), 283 - 7
Eight-year surveillance of non-albicans Candida spp . in an oncology department prior to and after fluconazole had been introduced into antifungal prophylaxis; Kunova A et al.; From 1989 until 1996, during the last 8 years, the proportion of Candida (C.) krusei, and other non-albicans Candida spp . isolated from surveillance cultures and from sterile body sites, was analyzed among 13,758 admissions in a National Cancer Institute . During these admissions a total of 9,042 isolates were prospectively collected from surveillance cultures, and 126 from blood cultures . The proportion of C . krusei among all organisms was 12.7% to 16.5% in 1989 through 1991, i.e., before fluconazole was introduced into prophylactic protocols . After the introduction of fluconazole into prophylaxis in acute leukemia in 1992 the incidence of C . krusei was 7.9% to 8.6% during 1994 to 1996 . After 5 years of using this drug for prophylaxis, the incidence of C . krusei was lower than before this drug was introduced in our institute . Among yeasts, the most frequently isolated pathogen was still Candida albicans (72.2% of all isolated fungal organisms) . Among molds, Aspergillus spp . was the most frequently isolated agent . Analyzing the etiology of proven fungal infections (fungemias) confirmed by positive blood cultures, C . albicans was the most common causative organism in 53.8% of cases . The incidence of fungemia due to Torulopsis (C.) glabrata and C . krusei before and after fluconazole introduction did not change . Of 126 organisms isolated from blood cultures, there was no increase in T . (C.) glabrata or C . krusei after introduction of fluconazole for prophylaxis and therapy, and the quoted 6.4% of fungemic episodes remained stable with an incidence of 1 fungemia/year since 1991 . The proportion of C . krusei and C . glabrata among Candida spp . was decreasing in our center between 1989 and 1996 . Also, the proportion of non-albicans Candida spp . among isolates decreased from 25.7% in 1990 to 11.9% in 1996.

Mycopathologia, 1997, 137(1), 1 - 8
Study of onychomycosis in Córdoba, Spain: prevailing fungi and pattern of infection; Velez A et al.; From a total of 20,004 patients seen during two years, we carried out a mycologic nail investigation (direct microscopy and repeated cultures) . Ninety-three (43.2%) of the nails were judged to be infected by their clinical appearance . They fulfilled the laboratory criteria required to start antifungal treatment (isolation of the same fungus in culture on two consecutive occasions), but only in 64 cases (29.7%) was there a clinical and mycological recovery once antifungal treatment and follow up were completed . yeasts were isolated in two thirds of the cases of onychomycosis, mainly from fingernails . Candida albicans, C . parapsilosis or both were the most prevalent species . Dermatophytes were found in 18.8% of the samples, especially from toenails . Trichophyton rubrum was the predominant species . Non-dermatophytic filamentous fungi were cultured in 17.2%, Scopulariopsis brevicaulis being the most prevalent species . The highest prevalence of onychomycosis was found in patients between 50 and 70 years of age . Females were affected more frequently than males . Fingernails were affected more frequently than toenails . Proximal subungual onychomycosis, secondary to paronychia (PSOp), was the most prevalent clinical type, although primary distal and lateral subungual onychomycosis (DLSO) and total dystrophic onychomycosis (TDO) were also frequent . PSOp was only observed in fingernails, while DLSO was almost only seen in toenails and TDO in both fingernails and toenails . All the clinical types were more frequent in women except TDO, which showed a similar prevalence in both sexes.

Cell, 1997 Sep 5, 90(5), 939 - 49
Nonfilamentous C . albicans mutants are avirulent; Lo HJ et al.; Candida albicans and Saccharomyces cerevisiae switch from a yeast to a filamentous form . In Saccharomyces, this switch is controlled by two regulatory proteins, Ste12p and Phd1p . Single-mutant strains, ste12/ste12 or phd1/phd1, are partially defective, whereas the ste12/ste12 phd1/phd1 double mutant is completely defective in filamentous growth and is noninvasive . The equivalent cph1/cph1 efg1/efg1 double mutant in Candida (Cph1p is the Ste12p homolog and Efg1p is the Phd1p homolog) is also defective in filamentous growth, unable to form hyphae or pseudohyphae in response to many stimuli, including serum or macrophages . This Candida cph1/cph1 efg1/efg1 double mutant, locked in the yeast form, is avirulent in a mouse model.

Acta Derm Venereol, 1997 Sep, 77(5), 388 - 91
Comparison of immune reactivity profiles against various environmental allergens between adult patients with atopic dermatitis and patients with allergic respiratory diseases; Matsumura N et al.; To clarify the pathomechanisms underlying the involvement of different organs by atopic dermatitis (AD) and allergic respiratory disease (ARD), we compared the immune reactivities to various environmental allergens between 46 adult patients who suffered only from AD but were without any history of ARD and 41 patients who had only ARD, using a RAST FEIA (radioallergosorbent test/fluoroenzyme immunoassay) and a scarification patch test . We also studied 42 healthy adult subjects in a similar fashion . Total serum IgE antibody levels were found to be far higher in the AD group than in the ARD and healthy control group, and RAST revealed that the AD group was sensitized to far larger numbers of allergens such as food mix, cereal mix, fungus mix and Candida albicans than were the other groups . The ARD group displayed a high incidence in RAST, comparable to that of the AD group, only against Japanese cedar and grass pollen mix antigen . However, the most remarkable difference in the immune reactivity profiles was that the AD group showed a uniquely higher RAST score and a lower incidence of positive patch test reactions to C . albicans antigen than did the ARD group . The reactivities in the ARD group to C . albicans antigen did not differ from those in the control group . Our present data suggest that a more pronounced shift from Th1 to Th2 cells, reactive against various allergens, takes place in AD patients.

Toxicol Lett, 1997 Jul 21, 92(2), 109 - 16
Evidence for arylamine N-acetyltransferase activity in the fungi Candida albicans; Fang SH et al.; N-acetyltransferase activities were determined in Candida albicans, which is a member of the normal flora of the mucous membranes in the respiratory, gastrointestinal and female genital tract . The N-acetylation of 2-aminofluorene and p-aminobenzoic acid by the N-acetyltransferase from Candida albicans was determined using high pressure liquid chromatography . The activities (mean +/- S.D.) of N-acetyltransferase from Candida albicans cytosols were 1.06 +/- 0.01 nmol/min per mg protein for the acetylation of 2-aminofluorene substrate, and not detectable levels of acetyl-p-aminobenzoic acid for the acetylation of p-aminobenzoic acid . The apparent kinetic constants Km and Vmax values were 0.17 +/- 0.06 mM and 1.43 +/- 0.42 nmol/min per mg protein, respectively, for 2-aminofluorene substrate . The optimum pH value for the enzyme activity was 8.0 . The optimal temperature for the enzyme activity is 40 degrees C for 2-aminofluorene substrate . Among a series of divalent cations and salts, Fe2+, SCN-, I-, and NH4+ were demonstrated to be the most potent inhibitors . The N-acetyltransferase activity was inhibited by iodoacetamide: at 0.25 mM iodoacetamide, activity was reduced 50% and 1.0 mM iodoacetamide inhibited activity more than 90% . This is the first demonstration of acetyl CoA arylamine N-acetyltransferase activity in the yeast-like fungus Candida albicans.

Curr Genet, 1997 Aug, 32(2), 108 - 14
The ornithine decarboxylase gene from Candida albicans . Sequence analysis and expression during dimorphism; Lopez MC et al.; The gene (CaODC) coding for ornithine decarboxylase, a key enzyme in polyamine biosynthesis, was cloned from Candida albicans by PCR and characterized . The deduced protein contains 470 amino acids together with the substrate- and co-factor-binding sequences which define the ornithine decarboxylases of eukaryotic species . It shows a high homology with other ODCs, mainly those from Saccharomyces cerevisiae and Neurospora crassa . A putative PEST sequence, which correlates quite well with those described for other fungal ODCs, could be identified in the protein . The mRNA of the gene is 2.4 kb in size and by primer extension a long leader sequence was found at -558 bases upstream of the predicted start of translation . An identical single ODC gene was identified in three different C . albicans strains . During the dimorphic switch, a transient initial increase in ODC activity was observed . This increase was not accompanied by a rise in the levels of the transcript, suggesting that ODC activity levels may be regulated post-transcriptionally during differentiation.

J Nihon Univ Sch Dent, 1997 Jun, 39(2), 67 - 70
Oral condition of patients with leukemia and lymphoma; Orbak R et al.; Different oral manifestations, the most prominent characteristics of the different oral symptoms, and oral colonization by Candida albicans were studied in 97 patients with leukemia and lymphoma . Oral manifestations usually occurred in both leukemias and lymphomas . The most common manifestation in both diseases was mucosal pallor . Oral colonization by Candida albicans in all patients was determined by the use of Sebouraud's agar plates and quantitative estimation of the colonization was made on a scale of +1 to +4 according to its presence in the four quadrants of the agar plates . Oral colonization by Candida albicans was between +1 and +4 in the patients with leukemia and was between +1 and +3 in the patients with lymphoma . Professional dental follow-up should be integrated into the medical follow-up . This is important not only for diagnosis of the disease, but also for prevention and treatment of complications during subsequent treatment.

Rev Inst Med Trop Sao Paulo, 1996 Nov-Dec, 38(6), 397 - 9
Candidin: comparison of two antigens for cutaneous delayed hypersensitivity testing; Fava-Netto C et al.; A candidin, which is a suspension of killed yeast cells, is commonly used for intradermal tests of delayed hypersensitivity, to evaluate the immunological cellular competence of the patient, when the test is applied along with other similar tests . When working with a cellular antigen, the histopathology of positive skin tests reveals a cellular infiltrate which not only presents a characteristic hypersensitivity reaction but also a neutrophilic abscess in the central part . This research presents the results of a comparison between the yeast cell suspension and the polysaccharide antigens, both obtained from the same strains of Candida albicans . The results obtained by skin tests in one hundred individuals were 61.0% with the polysaccharide antigen and 69.0% with the yeast cell suspension antigen . Concordant results concerning the two antigens were observed in 82.0% of the individuals . The discussion section presents an assumption to explain the differences of positivity obtained with the two antigens . We conclude that the polysaccharide antigen can be utilized in the intradermal test of delayed hypersensitivity to Candida albicans.

Appl Environ Microbiol, 1997 Sep, 63(9), 3488 - 93
Expression of a plant protein by Neurospora crassa; Rasmussen-Wilson SJ et al.; Heterologous expression of plant genes may serve as an important alternative for producing plant proteins . We have investigated the ability of the fungus Neurospora crassa to secrete zeamatin, a protein produced by Zea mays . Zeamatin was induced after being fused to glucoamylase, an extracellular hydrolase produced by N . crassa . Glucoamylase induction and other culture parameters were monitored in untransformed N . crassa grown in shaken liquid culture . A DNA plasmid, pGEZ, was constructed by inserting zeamatin-encoding cDNA into an expression cassette containing the promoter, a truncated open reading frame, and the terminator sequence of the N . crassa glucoamylase gene . Zeamatin-encoding cDNA was modified at the N terminus to include a kex-2 protease site, allowing cleavage of the chimeric product in the secretory pathway . Strains containing the chimeric gene construct were grown in liquid culture and induced for glucoamylase and zeamatin production . Zeamatin antibody detected a protein in a Western blot of concentrated culture supernatants that comigrated with authentic zeamatin . Secreted zeamatin was active in inhibiting the growth of Candida albicans in an agar diffusion assay, indicating that zeamatin had been correctly synthesized, processed, and secreted by N . crassa.

J Med Vet Mycol, 1997 Jul-Aug, 35(4), 289 - 93
Contact-sensing by hyphae of dermatophytic and saprophytic fungi; Perera TH et al.; Contact-sensing or thigmotropism is the directional growth response of cells in relation to topographical guidance cues . Thigmotropism is thought to play a major role in the location of infectable sites on plants by phytopathogenic fungi and has recently been shown to be a property of hyphae in the human pathogenic fungus Candida albicans . Here we show that hyphae of the dermatophytes Epidermophyton floccosum, Microsporum canis and Trichophyton mentagrophytes reorientate their direction of growth in response to grooves and pores of membrane substrata as did hyphae of the saprophytes Mucor mucedo and Neurospora crassa . This suggests that the thigmotropic behaviour of hyphae is not a specific property of pathogens, but rather a general feature of the growth of fungal hyphae that must forage for nutrients on surfaces and within solid materials.

J Med Microbiol, 1997 Sep, 46(9), 747 - 55
Release of Candida albicans yeast antigens upon interaction with human neutrophils in vitro; Ashley C et al.; Candida albicans is the leading cause of invasive candidosis . As conventional tests do not reliably detect invasive infection, attention has turned to the detection of C . albicans antigens circulating in blood . As antigen tests for invasive candidosis could be improved if C . albicans antigens released upon phagocytosis were defined, this study was undertaken to characterise antigens released during the interaction of yeasts and human neutrophils in vitro . An enzyme immunoassay developed previously to detect what were believed to be predominantly C . albicans cytoplasmic antigens in patients with invasive candidosis was used to follow the neutrophil-mediated release of yeast antigens . Serum opsonisation enhanced antigen release, which was rapid and essentially complete by 1 h . When fresh C . albicans yeasts were added to medium from cultures of neutrophils plus yeasts or neutrophils plus latex beads, additional yeast antigens were released . Medium from neutrophils plus yeasts or from yeasts alone had similar immunoblot patterns with rabbit antibodies to a C . albicans cytoplasmic antigen preparation, with the reactive antigens generally being of higher mol . wt than the reactive antigens in the antigen mixture used for preparation of the antiserum . The two supernates also had similar immunoblot patterns with rabbit anti-C . albicans cell-wall mannan antibodies . These results suggest that yeast surface antigens are released quickly during phagocytosis by neutrophils . Detection of such yeast surface antigens, possibly together with selected yeast cytoplasmic antigens, should improve the sensitivity of C . albicans antigen assays.

J Infect Dis, 1997 Sep, 176(3), 740 - 7
Antileukoprotease: an endogenous protein in the innate mucosal defense against fungi; Tomee JF et al.; Previous studies have suggested that endogenous protease inhibitors may participate in the mucosal host defense . Antileukoprotease (ALP) is an important protease inhibitor found on various mucosal surfaces, including those of the respiratory and genital tracts . This study reports on the antimicrobial activity of recombinant (r) ALP toward the human fungal pathogens Aspergillus fumigatus and Candida albicans . rALP expressed pronounced fungicidal activity toward metabolically active A . fumigatus conidia and C . albicans yeast cells; however, metabolically quiescent A . fumigatus conidia were totally resistant . In contrast with the protease inhibitory activity of rALP, the fungicidal activity was localized primarily in the NH2-terminal domain . On a molar base, the fungicidal activity of rALP was comparable with that of human defensins and lysozyme . In addition, rALP caused inhibition of C . albicans yeast cell growth . By exhibiting antifungal activity, ALP may play an important role in the innate mucosal defense against human pathogenic fungi.

Fungal Genet Biol, 1997 Jun, 21(3), 308 - 14
A Candida albicans genome project: cosmid contigs, physical mapping, and gene isolation; Tait E et al.; A new project to map the genome of the pathogenic fungus, Candida albicans, has been started . The entire genome was cloned as 5088 cosmids, stored in individual microtiter plate wells . DNA was prepared and fingerprinted using restriction digestion, fluorescent labeling, and analysis on an ABI sequencer . These data are being used to construct contigs of the genome . Simultaneously, a DNA pooling system has been set up, suitable for PCR-based isolation of cosmids containing any known gene . Ultimately, these approaches will lead to the creation of a physically based map of the C . albicans genome, providing the means to localize precisely all the genes, act as a substrate for genome sequencing projects, and provide probes for future studies of genome rearrangement and comparative genomics.

Curr Eye Res, 1997 Sep, 16(9), 930 - 5
Biodegradable scleral implant for intravitreal controlled release of fluconazole; Miyamoto H et al.; PURPOSE: To evaluate the feasibility of using a biodegradable polymeric scleral implant containing fluconazole (FLCZ), a bis-triazole antifungal agent, as a potential intravitreal-controlled drug delivery system . METHODS: The scleral implants, loaded with 10, 20, 30, and 50% FLCZ, were prepared with biodegradable polymers of poly (DL-lactide-co-glycolide) . Those with all loading doses were used for the in vitro release studies; those with 30% FLCZ were used for the intravitreal release studies in pigmented rabbits . The in vitro and in vivo release rates of FLCZ from the implants were measured periodically with spectrophotometry and high performance liquid chromatography, respectively . The effects of the implants on ocular tissues were evaluated ophthalmoscopically, histologically, and electrophysiologically . RESULTS: The scleral implants loaded with 10, 20, and 30% doses gradually released FLCZ over 4 weeks in vitro; those with 50% FLCZ released most of the drug in one week . FLCZ concentration in the rabbit vitreous remained within the 99% inhibitory concentration for Candida albicans for 3 weeks after implantation . The scleral implant gradually biodegraded, and it disappeared by 4 months after implantation . The electrophysiologic and histopathologic findings demonstrated no substantial toxic reactions in the ocular tissues . CONCLUSION: The current study suggests that a biodegradable, polymeric scleral implant containing FLCZ may be a promising intravitreal drug delivery system to treat fungal endophthalmitis.

Infect Immun, 1997 Sep, 65(9), 3822 - 7
Mannan-specific immunoglobulin G antibodies in normal human serum mediate classical pathway initiation of C3 binding to Candida albicans; Zhang MX et al.; Candida albicans activates both the classical and alternative complement pathways . Previous studies found that immunoglobulin G (IgG) in normal human serum (NHS) mediates classical pathway initiation . The goal of this study was to determine the role of candidal mannan-specific human IgG antibodies in complement activation . Mannan was purified from the yeast cells, and naturally occurring antimannan IgG was isolated from pooled NHS or plasma samples by immunoaffinity chromatography . Early activation and binding of C3, characteristics of classical pathway activity, were abolished in yeast- or mannan-absorbed serum but could be restored to absorbed serum with added purified antimannan IgG in a dose-dependent manner . Microscopically, the immunofluorescence pattern of initial C3 binding was diffuse over the entire cell surface for yeast cells incubated in NHS or in mannan-absorbed NHS supplemented with antimannan IgG but was asynchronous and focal for yeast cells incubated in EGTA-treated or mannan-absorbed NHS . The antimannan IgG level in serum samples from 21 donors varied from 17 to 570 microg/ml of serum compared to 220 microg in pooled NHS samples . The rate of initial C3 binding to yeast cells correlated with the level of antimannan IgG in sera from different individuals (r2 = 0.94) and could be accelerated in sera containing lower amounts of antimannan IgG with exogenous antimannan IgG . These observations identify antimannan IgG as the initiator of classical pathway C3 deposition on C . albicans . Given the variability in the levels of antimannan antibodies in sera from different individuals, the presence or absence of these antibodies may be an important determinant of host resistance to disseminated candidiasis.

Infect Immun, 1997 Sep, 65(9), 3539 - 46
A triple deletion of the secreted aspartyl proteinase genes SAP4, SAP5, and SAP6 of Candida albicans causes attenuated virulence; Sanglard D et al.; Secreted aspartyl proteinases (Saps) from Candida albicans are encoded by a multigene family with at least nine members (SAP1 to SAP9) and are considered putative virulence factors important for the pathogenicity of this human pathogen . The role of Sap isoenzymes in the virulence of C . albicans has not yet been clearly established, and therefore, using recent progress in the genetics of this yeast, we have constructed a panel of isogenic yeasts, each with a disruption of one or several SAP genes . We focused on the construction of a C . albicans strain in which three related SAP genes (SAP4, SAP5, and SAP6) were disrupted . Growth of the delta sap4,5,6 triple homozygous null mutant DSY459 in complex medium was not affected, whereas, interestingly, growth in a medium containing protein as the sole nitrogen source was severely impaired compared to the growth of the wild-type parent strain SC5314 . Since the presence of Sap2 is required for optimal growth on such medium, this suggests that Sap4, Sap5, or Sap6 plays an important role for the process of induction of SAP2 . When guinea pigs and mice were injected intravenously with DSY459, their survival time was significantly longer than that of control animals infected with the wild-type SC5314 . Attenuated virulence of DSY459 was followed by a significant reduction of yeast cells in infected organs . These data suggest that the group of Sap4, Sap5, and Sap6 isoenzymes is important for the normal progression of systemic infection by C . albicans in animals.

Infect Immun, 1997 Sep, 65(9), 3529 - 38
Disruption of each of the secreted aspartyl proteinase genes SAP1, SAP2, and SAP3 of Candida albicans attenuates virulence; Hube B et al.; Secreted aspartyl proteinases (Saps), encoded by a gene family with at least nine members (SAP1 to SAP9), are one of the most discussed virulence factors produced by the human pathogen Candida albicans . In order to study the role of each Sap isoenzyme in pathogenicity, we have constructed strains which harbor mutations at selected SAP genes . SAP1, SAP2, and SAP3, which are regulated differentially in vitro, were mutated by targeted gene disruption . The growth rates of all homozygous null mutants were similar to those of the isogenic wild-type parental strain (SC5314) in complex and defined media . In medium with protein as the sole source of nitrogen, sap1 and sap3 mutants grew with reduced growth rates but reached optical densities similar to those measured for SC5314 . In contrast, sap2 null mutants tended to clump, grew poorly in this medium, and produced the lowest proteolytic activity . Addition of ammonium ions reversed such growth defects . These results support the view that Sap2 is the dominant isoenzyme . When sap1, sap2, and sap3 mutants were injected intravenously in guinea pigs and mice, the animals had increased survival rates compared to those of control animals infected with SC5314 . However, reduction of proteolytic activity in vitro did not correlate directly with the extent of attenuation of virulence observed for all Sap-deficient mutants . These data suggest that SAP1, SAP2, and SAP3 all contribute to the overall virulence of C . albicans and presumably all play important roles during disseminated infections.

J Clin Microbiol, 1997 Sep, 35(9), 2348 - 58
Parity among the randomly amplified polymorphic DNA method, multilocus enzyme electrophoresis, and Southern blot hybridization with the moderately repetitive DNA probe Ca3 for fingerprinting Candida albicans; Pujol C et al.; Randomly amplified polymorphic DNA (RAPD) analysis, multilocus enzyme electrophoresis (MLEE), and Southern blot hybridization with moderately repetitive DNA probes have emerged as effective fingerprinting methods for the infectious fungus Candida albicans . The three methods have been compared for their capacities to identify identical or highly related isolates, to cluster weakly related isolates, to discriminate between unrelated isolates, and to assess microevolution within a strain . By computing similarity coefficients between 29 isolates from three cities within the continental United States, strong concordance of the results is demonstrated for RAPD analysis, MLEE, and Southern blot hybridization with the moderately repetitive probe Ca3, and weaker concordance of the results is demonstrated for these three fingerprinting methods and Southern blot hybridization with the moderately repetitive probe CARE2 . All methods were also demonstrated to be able to resolve microevolution within a strain, with the Ca3 probe exhibiting the greatest resolving power . The strong correlations demonstrated between polymorphic markers assessed by the four independent fingerprinting methods and the nonrandom association between loci demonstrated by RAPD analysis and MLEE provide evidence for strong linkage disequilibrium and a clonal population structure for C . albicans . In addition, a synapomorphic allele, Pep-3A, was found to be present in all members of one of the three clusters discriminated by RAPD analysis, MLEE, and Ca3 fingerprinting, supporting the concordance of the clustering capacities of the three methods, the robustness of the clusters, and the clonal nature of the clusters.

J Clin Microbiol, 1997 Sep, 35(9), 2320 - 4
Rapid flow cytometric susceptibility testing of Candida albicans; Ramani R et al.; A rapid flow cytometric assay for in vitro antifungal drug susceptibility testing was developed by adapting the proposed reference method for broth macrodilution testing of yeasts . Membrane permeability changes caused by the antifungal agent were measured by flow cytometry using propidium iodide, a nucleic acid-binding fluorochrome largely excluded by the intact cell membrane . We determined the in vitro susceptibility of 31 Candida albicans isolates and two quality control strains (Candida parapsilosis ATCC 22019 and Candida krusei ATCC 6258) to amphotericin B and fluconazole . Amphotericin B MICs ranged from 0.03 to 2.0 microg/ml, while fluconazole MICs ranged from 0.125 to 128 microg/ml . This method results in clear-cut endpoints that were reproducible . Four-hour incubation was required for fluconazole, whereas a 2-h incubation was sufficient for amphotericin B to provide MICs comparable to the reference macrodilution method developed by the National Committee for Clinical Laboratory Standards Subcommittee on Antifungal Susceptibility Tests . Results of these studies show that flow cytometry provides a rapid and sensitive in vitro method for antifungal susceptibility testing of C . albicans.

J Dent, 1997 Sep, 25(5), 373 - 7
Prolonged antimicrobial effect of tissue conditioners containing silver-zeolite; Matsuura T et al.; OBJECTIVES: The purpose of this study was to elucidate the in vitro antimicrobial effect of tissue conditioners containing silver-zeolite on Candida albicans and nosocomial respiratory infection-causing bacteria, Staphylococcus aureus and Pseudomonas aeruginosa . METHODS: Five commercially available tissue conditioners were selected: Visco-gel (VG), GC Soft-Liner (SL), Fitt (FT), SR-Ivoseal (IV) and Shofu Tissue Conditioner (TC) . Samples, 10 x 10 x 2.5 mm in size, contained silver-zeolite (SZ sample) and no SZ (N sample) . The antimicrobial effects of these two samples were evaluated as a percentage of viable cells (CFU) in a microbial suspension (100 microliter) in phosphate-buffered saline with or without immersion in artificial saliva for four weeks . The borderline of the antimicrobial effect was determined at 0.1% viable cells . RESULTS: With the SZ samples, all tested microbes were killed under both conditions of no immersion and immersion in saliva . In non-immersed N samples, however, no cells of C . albicans (except with VG) and S . aureus survived, whereas the percentage of viable cells of P . aeruginosa was similar to that found in the control . However, with immersion in saliva, viable cells of C . albicans in some N samples (VG, SL and TC) increased compared with non-immersion samples by more than 0.1% . CONCLUSION: Tissue conditioners containing SZ have been shown to have antimicrobial effects for four weeks on C . albicans and nosocomial respiratory infection-causing bacteria in saliva in vitro.

FEMS Microbiol Lett, 1997 Aug 15, 153(2), 349 - 55
Degradation of human subendothelial extracellular matrix by proteinase-secreting Candida albicans; Morschhauser J et al.; Candida albicans infections in severely immunocompromized patients are not confined to mucosal surfaces; instead the fungus can invade through epithelial and endothelial layers into the bloodstream and spread to other organs, causing disseminated infections with often fatal outcome . We investigated whether secretion of the C . albicans acid proteinase facilitates invasion into deeper tissues by degrading the subendothelial basement membrane . After cultivation under conditions that induce the secretion of the acid proteinase, C . albicans degraded radioactively metabolically labeled extracellular matrix proteins from a human endothelial cell line . The degradation was inhibited in the presence of pepstatin A, an inhibitor of acid proteinases . Pepstatin A-sensitive degradation of the soluble and immobilized extracellular matrix proteins fibronectin and laminin by proteinase-producing C . albicans was also detected, whereas no degradation was observed when the expression of the acid proteinase was repressed . Our results demonstrate that the C . albicans acid proteinase degrades human subendothelial extracellular matrix; this may be of importance in the penetration of C . albicans into circulation and deep organs.

Arch Biochem Biophys, 1997 Aug 15, 344(2), 350 - 6
Ingestion of Candida albicans down-regulates mannose receptor expression on rat macrophages; Shepherd VL et al.; The frequency of infection and death due to various Candida species has increased steadily during the past decade, with mucocutaneous candidal infections as a common problem in the immunocompromised host . Mononuclear phagocytes are important in phagocytosis of this organism . In areas where there are low levels of opsonins, the macrophage-specific mannose receptor plays a dominant role in mediating Candida albicans ingestion . Following receptor-mediated infection, the host macrophage produces inflammatory cytokines and mediators that lead to ultimate killing of the invading Candida . Infection of macrophages by pathogens often leads to altered function that might effect their subsequent host defense properties . For example, function of both the complement receptor type 3 and the mannose receptor are down-regulated following exposure to pathogens or pathogen-derived products . In the current study, we have examined the down-regulation of mannose receptor expression following Candida infection and have investigated possible mechanisms that might be involved . Mannose receptor activity was decreased following 24 h postinfection with Candida . Both tumor necrosis factor and nitric oxide were produced during the infection, and inhibition of the these mediators partially blocked the effect on the receptor . Infection with Candida also inhibited the ability of dexamethasone to up-regulate mannose receptor expression . Finally, mannose receptor protein turnover was accelerated in Candida-infected macrophages . We conclude that Candida down-regulates one of the receptors involved in its internalization through a combination of production of modulatory molecules and enhanced receptor degradation . These results support the hypothesis that pathogens that infect macrophages have the ability to alter the phagocytic pathways available for subsequent host defense.

J Periodontol, 1997 Aug, 68(8), 729 - 33
In vitro antifungal properties of mouthrinses containing antimicrobial agents; Giuliana G et al.; The purpose of this study was to investigate the in vitro antifungal properties of seven commercial mouthrinses containing antimicrobial agents . These included cetylpyridinium chloride (CPC), chlorhexidine digluconate (CHX), hexetidine (HEX), sanguinarine (SNG), and triclosan (TRN) . The minimum fungicidal concentration (MFC) against six species of yeasts was determined by a broth macrodilution method . The kill-time of mouthrinses at half the concentration of the commercial formulations was also determined . MFCs were achieved with each mouthrinse, except the SNG-containing mouthrinse, against all the organisms being tested . However, the CPC-containing mouthrinse appeared more active than the other products (P < 0.001) . There were no significant differences in MFC values among CHX mouthrinse products, once adjusted for initial concentration differences (P = 0.1) . Kill-times of mouthrinses containing either CHX or CPC were less than or equal to 180 seconds with all the species of yeasts, and no significant differences were found among these products (P = 0.18) . On the other hand, mouthrinses containing either TRN or HEX did not show a lethal effect on Candida albicans, Candida parapsilosis, or Candida guilliermondii . No kill-times were achieved with the SNG-containing mouthrinse . These results suggest that mouthrinses containing antimicrobial agents might represent an appropriate alternative to conventional antifungal drugs in the management of oral candidiasis . However, the effectiveness of antimicrobial mouthrinses as antifungal agents needs to be evaluated in further clinical trials.

AJNR Am J Neuroradiol, 1997 Aug, 18(7), 1303 - 6
Disseminated miliary cerebral candidiasis; Lai PH et al.; We present a case of disseminated intracranial infection by Candida albicans in a 5-year-old girl who had fever and a change of consciousness after surgery for complex congenital heart malformation . MR imaging revealed multiple small ring-enhancing hemorrhagic abscesses . One year after antifungal treatment, the abscesses and ventriculomegaly were almost completely resolved . The patient was discharged in a stable but vegetative condition.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1997 Aug, 84(2), 149 - 53
Candidal colonization and oral candidiasis in patients undergoing oral and pharyngeal radiation therapy; Ramirez-Amador V et al.; OBJECTIVES: Radiotherapy-induced hyposalivation encourages oral candidal colonization that often leads to oral/pharyngeal candidiasis . The purpose of this study was to quantitate oral candidal colonization, assess signs, symptoms, and response to antifungal management, speciate Candida, and evaluate the influence of smoking and dentures . STUDY DESIGN: Forty-six patients undergoing radiation therapy for oral/pharyngeal squamous cell carcinoma were evaluated clinically and by Candidal cultures before, during, and after irradiation . RESULTS: All patients complained of progressive xerostomia . There was a significant increase in the prevalence of positive candidal cultures (P = < 0.0001): baseline 43%, completion of radiotherapy 62%, and follow-up timepoint 75% . Smoking and denture wearing were not statistically significant risk factors for increased candidal colonization (p = 0.085 and p = 0.420, respectively) . Eight patients developed clinical candidiasis . Although five responded clinically to systemic antifungal medication, all follow-up cultures remained positive . Candida albicans was the predominant species at baseline and completion of radiation (85% and 68%, respectively) . CONCLUSIONS: When salivary glands are included in the field of radiation, xerostomia occurs, causing progressive increases in oral Candida colonization . Because 17.4% developed clinical candidiasis during radiotherapy and the question of fungal resistance remains speculative, a recommendation for the prophylactic use of antifungal medication is unresolved.

J Bacteriol, 1997 Aug, 179(16), 5030 - 6
Evidence that part of a centromeric DNA region induces pseudohyphal growth in a dimorphic yeast, Candida maltosa; Nakazawa T et al.; We observed that a YCp-type vector having the centromeric DNA (CEN) sequence previously isolated from the genome, but not a YRp-type vector lacking the CEN sequence, induced pseudohyphal growth in a dimorphic fungi, Candida maltosa, which had been shown to be closely related to Candida albicans by phylogenetic analysis . Deletion analysis of the CEN sequence revealed that the intact CEN sequence was not required for the induction, but part of it, having partial centromeric activity, was enough for the induction . By screening the gene library of this yeast for the sequences which induced pseudohyphal growth, we isolated three different DNA fragments which also had part of the centromere-like sequence . Partial centromeric activity of these fragments was confirmed by three criteria: low copy number and high stability of the plasmids carrying these fragments and rearrangement at high frequency of the plasmid DNA with one of these fragments plus the CEN sequence . Furthermore, when the GGTAGCG sequence commonly found in one copy in each of these four sequences was mutated in the CEN sequence by site-directed mutagenesis, both partial centromeric activity and pseudohyphal growth-inducing activity of the CEN sequence were lost . These results indicated that part of CEN region with partial centromeric activity induces pseudohyphal growth in C . maltosa . It is suggested that some cellular components which interact with the sequence containing GGTAGCG required for centromeric activity are involved in the regulation of the transition between yeast forms and pseudohyphal forms of the cells.

J Bacteriol, 1997 Aug, 179(16), 4992 - 9
The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase of Candida albicans is a surface antigen; Gil-Navarro I et al.; A lambda gt11 cDNA library from Candida albicans ATCC 26555 was screened by using pooled sera from two patients with systemic candidiasis and five neutropenic patients with high levels of anti-C . albicans immunoglobulin M antibodies . Seven clones were isolated from 60,000 recombinant phages . The most reactive one contained a 0.9-kb cDNA encoding a polypeptide immunoreactive only with sera from patients with systemic candidiasis . The whole gene was isolated from a genomic library by using the cDNA as a probe . The nucleotide sequence of the coding region showed homology (78 to 79%) to the Saccharomyces cerevisiae TDH1 to TDH3 genes coding for glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and their amino acid sequences showed 76% identity; thus, this gene has been named C . albicans TDH1 . A rabbit polyclonal antiserum against the purified cytosolic C . albicans GAPDH (polyclonal antibody {PAb} anti-CA-GAPDH) was used to identify the GAPDH in the beta-mercaptoethanol extracts containing cell wall moieties . Indirect immunofluorescence demonstrated the presence of GAPDH at the C . albicans cell surface, particularly on the blastoconidia . Semiquantitative flow cytometry analysis showed the sensitivity of this GAPDH form to trypsin and its resistance to be removed with 2 M NaCl or 2% sodium dodecyl sulfate . The decrease in fluorescence in the presence of soluble GAPDH indicates the specificity of the labelling . In addition, a dose-dependent GAPDH enzymatic activity was detected in intact blastoconidia and germ tube cells . This activity was reduced by pretreatment of the cells with trypsin, formaldehyde, and PAb anti-CA-GAPDH . These observations indicate that an immunogenic, enzymatically active cell wall-associated form of the glycolytic enzyme GAPDH is found at the cell surface of C . albicans cells.

Curr Biol, 1997 Aug 1, 7(8), 539 - 46
Virulence and hyphal formation of Candida albicans require the Ste20p-like protein kinase CaCla4p; Leberer E et al.; BACKGROUND: The pathogenic fungus Candida albicans is capable of a morphological transition from a unicellular budding yeast to a filamentous form . Extensive filamentous growth leads to the formation of mycelia displaying hyphae with branches and lateral buds . Hyphae have been observed to adhere to and invade host tissues more readily than the yeast form, suggesting that filamentous growth may contribute to the virulence of this major human pathogen . A molecular and genetic understanding of the potential role of morphological switching in the pathogenicity of C . albicans would be of significant benefit in view of the increasing incidence of candidiasis . RESULTS: The CaCLA4 gene of C . albicans was cloned by functional complementation of the growth defect of cells of the budding yeast Saccharomyces cerevisiae deleted for the STE20 gene and the CLA4 gene . CaCLA4 encodes a member of the Ste20p family of serine/threonine protein kinases and is characterized by a pleckstrin homology domain and a Cdc42p-binding domain in its amino-terminal non-catalytic region . Deletion of both alleles of CaCLA4 in C . albicans caused defects in hyphal formation in vitro, in both synthetic liquid and solid media, and in vivo in a mouse model for systemic candidiasis . The gene deletions reduced colonization of the kidneys in infected mice and suppressed C . albicans virulence in the mouse model . CONCLUSIONS: Our results demonstrate that the function of the CaCla4p protein kinase is essential for virulence and morphological switching of C . albicans in a mouse model . Thus, hyphal formation of C . albicans mediated by CaCla4p may contribute to the pathogenicity of this dimorphic fungus, suggesting that regulators of morphological switching may be useful targets for antifungal drugs.

Antimicrob Agents Chemother, 1997 Aug, 41(8), 1812 - 4
In vitro activities of terbinafine in combination with fluconazole and itraconazole against isolates of Candida albicans with reduced susceptibility to azoles; Barchiesi F et al.; A checkerboard microdilution method was applied to study the in vitro interaction of terbinafine with either fluconazole and itraconazole against 30 strains of Candida albicans . Synergy was observed in 40% of the terbinafine-fluconazole interactions and in 43% of the terbinafine-itraconazole interactions, while antagonism was not observed . Even when only additivity was achieved, the combinations still showed beneficial effects since at least twofold reductions in the MICs of both drugs were found in 100% of the terbinafine-fluconazole interactions and in 76% of the terbinafine-itraconazole interactions.

Antimicrob Agents Chemother, 1997 Aug, 41(8), 1775 - 7
Treatment of murine disseminated candidiasis with L-743,872; Graybill JR et al.; L-743,872 (M991), which is a pneumocandin derivative, was evaluated in a mouse model of disseminated candidiasis caused by a fluconazole-resistant isolate of Candida albicans . In immunocompetent mice M991 prolonged survival at doses as low as 0.0125 mg/kg of body weight per day . In neutropenic mice 0.05 mg/kg was the lowest effective dose . M991 is a very potent drug for treatment of disseminated candidiasis.

Antimicrob Agents Chemother, 1997 Aug, 41(8), 1625 - 35
Isolation of Candida species on media with and without added fluconazole reveals high variability in relative growth susceptibility phenotypes; Schoofs A et al.; Mouthwashes from human immunodeficiency virus-positive individuals were sampled for yeasts by direct plating on a differential agar medium with and without added fluconazole and via enrichment broths with and without added fluconazole . The colonies of the yeasts isolated were tested for relative growth in the presence of single concentrations of itraconazole and fluconazole . Among 258 culture plates containing yeasts obtained via different isolation routes from 86 yeast-positive samples, 33 (12.7%) of the plates showed unexpectedly high colony-to-colony variation in relative growth . Intercolony variation was seen in 41 (47.7%) of the 86 isolates when relative growth data were analyzed for all colonies of an isolate tested, regardless of the medium used for isolation . The prevalence of relative growth variability with the azoles was highest for Candida glabrata (100% of 13 isolates), followed by Candida krusei (60% of 5 isolates) and Candida albicans (40% of 53 isolates), and the visual patterns of variability seen in scatter plots of the data showed species specificity . Relative growth phenotypes generally tended to be stable for each yeast colony in subcultures, whether or not the medium used for subculture contained antifungal agents . DNA fingerprinting of stable and variable C . albicans isolates showed changes in band patterns detected with the probe Ca3, suggesting that the variability may have resulted from selection of different subtypes of the yeasts during the isolation procedure . These findings suggest that the yeasts isolated from single clinical samples were often not clonal in nature . The relative growth test revealed colony variability more readily than conventional susceptibility testing.

J Bacteriol, 1997 Aug, 179(15), 4654 - 63
Cloning and characterization of CSP37, a novel gene encoding a putative membrane protein of Candida albicans; Sentandreu M et al.; In the course of an analysis of the functions and assembly of the cell wall of Candida albicans, we have cloned and characterized a gene, which we designated CSP37 (cell surface protein), encoding a 37-kDa polypeptide which is a membrane-associated protein . The gene was isolated by immunological screening of a DNA library constructed from mycelial cells with a polyclonal serum raised against cell walls of this morphology . Analysis of the nucleotide sequence of a corresponding genomic DNA fragment revealed a single open reading frame which encodes a predicted protein of 321 amino acids with no significant homology to others in the databases . Disruption of the CSP37 gene by the method described by Fonzi and Irwin (Genetics 134:717-728, 1993) eliminated expression of the Csp37 protein . The mutant strains showed no apparent defect in cell viability, growth, or cell wall assembly but displayed attenuated virulence in systemic infections induced in mice and reduced the ability to adhere to polystyrene.

Obstet Gynecol, 1997 Aug, 90(2), 252 - 6
Immune compromise and prevalence of Candida vulvovaginitis in human immunodeficiency virus-infected women; Duerr A et al.; OBJECTIVE: To investigate the effect of human immunodeficiency virus (HIV) infection on vaginal yeast colonization and symptomatic vulvovaginitis and to explore the effects of immune compromise on these conditions in HIV-positive women . METHODS: Between September 1991 and May 1993, 223 HIV-positive women without AIDS-defining conditions were enrolled for prospective follow-up and compared with 289 HIV-negative women enrolled in a concurrent study . Standardized gynecologic assessment was carried out . RESULTS: Cultures from 81 of 223 (36%) HIV-positive women and 72 of 289 (25%) HIV-negative women were positive for any yeast . The most commonly isolated yeasts were Candida albicans and Torulopsis glabrata; the proportion of non-C albicans isolates (26%) did not differ by serostatus . The rates of C albicans colonization and vulvovaginitis among immunocompetent (CD4 count at least 500 cells/mm3) HIV-positive women did not differ from those among HIV-negative women . Among HIV-positive women, risks for colonization and for symptomatic vulvovaginitis were increased approximately threefold and fourfold respectively, in women with CD4 counts below 200 cells/mm3 compared with either immunocompetent HIV-positive women or HIV-negative women . CONCLUSION: The yeast species isolated from HIV-positive and HIV-negative women were similar . Rates of vaginal colonization and vaginitis were similar among nonimmunocompromised HIV-positive women and HIV-negative women . Elevated rates of yeast colonization and vaginitis were not seen among this population of HIV-infected women before immune compromise . Both vaginal colonization and symptomatic vaginitis increased with immune compromise among HIV-positive women, especially at CD4 counts below 200 cells/mm3.

J Infect Dis, 1997 Aug, 176(2), 492 - 8
Human immunodeficiency virus type 1 gp41 binds to Candida albicans via complement C3-like regions; Wurzner R et al.; Oral candidiasis in human immunodeficiency virus type 1 (HIV-1)-infected persons is believed to be caused by the acquired T lymphocyte immunodeficiency . The direct interaction of C . albicans and HIV-1 in vitro was investigated . Twice as many yeasts adhered to cells transfected with the HIV-1 env gene as they did to controls . HIV-1 rsgp160 and rsgp41 but not rsgp120 were found to bind to Candida albicans via two C3-like regions within gp41 . Normal human serum, but not C3-depleted serum, was able to inhibit rsgp41 binding to C . albicans . Vice versa, rsgp160 and rsgp41 were able to block rosetting of C . albicans with iC3b-coated sheep erythrocytes . Binding to C . albicans, and its inhibition by rsgp41 or rsgp160, was confirmed for the whole virus . Therefore, oral candidiasis in HIV-1-infected subjects may be augmented or may even be initiated by direct interaction between C . albicans and HIV-1 or HIV-1-infected cells.

J Biol Chem, 1997 Aug 1, 272(31), 19304 - 13
AP1-mediated multidrug resistance in Saccharomyces cerevisiae requires FLR1 encoding a transporter of the major facilitator superfamily; Alarco AM et al.; We have isolated a Candida albicans gene that confers resistance to the azole derivative fluconazole (FCZ) when overexpressed in Saccharomyces cerevisiae . This gene encodes a protein highly homologous to S . cerevisiae yAP-1, a bZip transcription factor known to mediate cellular resistance to toxicants such as cycloheximide (CYH), 4-nitroquinoline N-oxide (4-NQO), cadmium, and hydrogen peroxide . The gene was named CAP1, for C . albicans AP-1 . Cap1 and yAP-1 are functional homologues, since CAP1 expression in a yap1 mutant strain partially restores the ability of the cells to grow on toxic concentrations of cadmium or hydrogen peroxide . We have found that the expression of YBR008c, an open reading frame identified in the yeast genome sequencing project and predicted to code for a multidrug transporter of the major facilitator superfamily, is dramatically induced in S . cerevisiae cells overexpressing CAP1 . Overexpression of either CAP1 or YAP1 in a wild-type strain results in resistance to FCZ, CYH, and 4-NQO, whereas such resistance is completely abrogated (FCZ and CYH) or strongly reduced (4-NQO) in a ybr008c deletion mutant, demonstrating that YBR008c is involved in YAP1- and CAP1-mediated multidrug resistance . YBR008c has been renamed FLR1, for fluconazole resistance 1 . The expression of an FLR1-lacZ reporter construct is strongly induced by the overexpression of either CAP1 or YAP1, indicating that the FLR1 gene is transcriptionally regulated by the Cap1 and yAP-1 proteins . Taken collectively, our results demonstrate that FLR1 represents a new YAP1-controlled multidrug resistance molecular determinant in S . cerevisiae . A similar detoxification pathway is also likely to operate in C . albicans.

Infect Immun, 1997 Aug, 65(8), 3493 - 5
Effect of thermal injury on the adherence of Candida albicans to murine splenic tissue; Neely AN et al.; In a mouse model of thermal injury, an increase in burn size produced a decrease in the ratio of Candida albicans cells adherent to the marginal zone to those adherent to the white pulp of the spleen, an increase in the number of Candida cells in the circulation and in the kidneys, and an increase in mortality.

Infect Immun, 1997 Aug, 65(8), 3399 - 405
Protective role of antimannan and anti-aspartyl proteinase antibodies in an experimental model of Candida albicans vaginitis in rats; De Bernardis F et al.; The role of antibodies (Abs) in the resistance to vaginal infection by Candida albicans was investigated by using a rat vaginitis model . Animals receiving antimannoprotein (anti-MP) and anti-aspartyl proteinase (Sap) Ab-containing vaginal fluids from rats clearing a primary C . albicans infection showed a highly significant level of protection against vaginitis compared to animals given Ab-free vaginal fluid from noninfected rats . Preabsorption of the Ab-containing fluids with either one or both proteins MP and Sap sequentially reduced or abolished, respectively, the level of protection . A degree of protection against vaginitis was also conferred by postinfectious administration of anti-Sap and anti-MP monoclonal antibodies (provided the latter were directed against mannan rather than protein epitopes of MP) and by intravaginal immunization with a highly purified, polysaccharide-free Sap preparation . Postinfectious administration of pepstatin A, a potent Sap inhibitor, greatly accelerated the clearance of C . albicans from rat vagina . No anti-MP or anti-Sap Abs were elicited during a C . albicans vaginal infection of congenitally athymic nude rats . Although they were as able as their euthymic counterparts to clear the primary infection, these animals did not show increased resistance to a rechallenge, demonstrating that induction of anticandidal protection in normal rats was a thymus-dependent Ab response . Overall, our data strengthen the concept that Abs against some defined Candida antigens are relevant in the mechanism of acquired anticandidal protection in vaginitis . The T-cell dependence of this protection may also provide a link between cell-mediated and humoral immunity in vaginal infection.

Transplantation, 1997 Jul 15, 64(1), 66 - 73
Prophylaxis of cytomegalovirus infection in liver transplantation: a randomized trial comparing a combination of ganciclovir and acyclovir to acyclovir . NIDDK Liver Transplantation Database; Badley AD et al.; BACKGROUND: The optimal prophylactic regimen to prevent cytomegalovirus (CMV) infection and disease in orthotopic liver-transplant patients remains to be established . We tested whether a combination of intravenous ganciclovir (GCV) followed by high dosages of oral acyclovir (ACV) for 4 months provided a higher degree of protection from CMV than oral ACV alone . METHODS: One hundred sixty-seven liver-transplant recipients were randomized to receive 120 days of antiviral treatment starting at the time of transplantation consisting of either ACV 800 mg orally four times daily (n=84) or 14 days of GCV 5 mg/kg intravenously every 12 hr followed by oral ACV 800 mg four times daily (n=83) . Prospective laboratory and clinical surveillance was performed to determine primary endpoints (onset of CMV infection and CMV disease) and secondary endpoints (rates of fungal and bacterial infection, allograft rejection, and survival after transplantation) . One-year event rates are presented as cumulative percentages . RESULTS: During the first year after transplantation, CMV infection developed in 57% of patients treated with ACV and in 37% of patients treated with GCV + ACV (P=0.001) . CMV disease developed in 23% of patients treated with ACV and in 11% of patients treated with GCV + ACV (P=0.03) . In seronegative recipients of allografts from CMV-seropositive donors (D+/R-), CMV disease developed in 58% of patients treated with ACV and in 25% of patients treated with GCV + ACV (P=0.04) . In the D+/R- group, 54% of patients treated with ACV and 17% of patients treated with GCV + ACV developed infection with Candida albicans (P=0.05) . CONCLUSIONS: Prophylaxis of CMV infection in liver-transplant patients with 14 days of intravenous GCV followed by high-dosage oral ACV is more effective than high-dosage oral ACV alone at reducing CMV infection and disease, even for patients in the D+/R- CMV serological group.

AIDS, 1997 Jul 15, 11(9), 1095 - 101
Oral transmission of Candida albicans between partners in HIV-infected couples could contribute to dissemination of fluconazole-resistant isolates; Dromer F et al.; BACKGROUND: Fluconazole resistance has emerged among Candida albicans isolates and has been associated with the prolonged or repeated use of the drug . This study was designed to discover whether transmission of oral isolates could occur between sexual partners and thereby explain fluconazole resistance in patients never treated with the drug . MATERIALS AND METHODS: The oral flora of 10 HIV-infected couples (five heterosexual and five homosexual) were studied . In vitro susceptibility testing and genotyping (restriction fragment length polymorphism with EcoRI and HinfI) were used to delineate strain relatedness for 230 clones (five clones per sample, one to four samples per patient) . RESULTS: The genetic diversity of the clones with one DNA subtype was specific to a given patient or a given couple, except in one case in which unrelated patients shared clones of the same genotype . The persistence of clones between partners was stable over time in six out of 10 couples and only transient in one couple . Fluconazole resistance in isolates from patients who had never been treated with azoles was associated in three patients with the first episode of oropharyngeal candidiasis and treatment failure . CONCLUSION: The observation that couples tended to share genetically indistinguishable clones was highly suggestive of transmission between partners . This phenomenon may, in part, explain the pathogenesis of oropharyngeal candidiasis and the increased frequency of fluconazole resistance both in vitro and in vivo.

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