Candida albicans

Int J Immunopharmacol, 1999 Mar, 21(3), 161 - 76
Atiprimod (SK&F 106615), a novel macrophage targeting agent, enhances alveolar macrophage candidacidal activity and is not immunosuppressive in Candida-infected mice; Badger AM et al.; Azaspiranes are novel macrophage-targeting agents with activity in preclinical animal models of autoimmune disease and transplantation . The purpose of this work was to determine the effects of atiprimod (SK&F 106615), an azaspirane being developed for the treatment of rheumatoid arthritis, on rat pulmonary alveolar macrophage (AM) function and immunocompetance in Candida-infected mice . AM from rats treated with 20 mg/kg/day of atiprimod for 15 days demonstrated enhanced killing of Candida albicans ex vivo . Concentration-dependent increases in candidacidal activity were also observed as early as one hour after exposure in vitro in AM from untreated normal rats . Treatment of AM with atiprimod in vitro did not increase particulate-stimulated superoxide production or phagocytosis of Candida but decreased their ability to concentrate acridine orange, indicating an increase in lysosomal pH . Increased candidacidal activity was inhibited by superoxide dismutase and catalase, suggesting a role for reactive oxygen intermediates (ROI) . Atiprimod also increased free radical-mediated killing of Candida in the presence of H2O2, iron and iodide in a cell-free system . These findings indicated that treatment with atiprimod increased the candidacidal activity of rat AM in a free radical-dependent manner . The data also suggested that atiprimod did not increase ROI production by AM, but rather increased the efficiency of radical-mediated killing . This increase may be caused by cyclization of atiprimod, facilitating electron transfer and peroxidation of lipid membranes . In vivo studies in Candida-infected CBA mice showed that atiprimod (10 mg/kg/day), did not compromise immune function in the infected mice and could be differentiated from prototypical immunosuppressive compounds used for treatment of autoimmune diseases.

J Antibiot (Tokyo), 1999 Mar, 52(3), 311 - 8
Aspirochlorine: a highly selective and potent inhibitor of fungal protein synthesis; Monti F et al.; Aspirochlorine, a compound belonging to the gliotoxin family of compounds, exhibits antifungal and antibacterial activity but its mechanism of action remains unknown . In this study we show that aspirochlorine inhibits the pathogenic fungus Candida albicans by acting on fungal protein synthesis . The compound selectively inhibits cell-free protein synthesis when using a C . albicans system, but does not inhibit this synthesis in vitro when tested with bacterial and mammalian systems . Moreover, in intact C . albicans cells, aspirochlorine inhibits protein synthesis but does not inhibit chitin, DNA or glucan synthesis though at high concentrations some inhibition of RNA synthesis is observed . By contrast, in intact Bacillus subtilis cells, aspirochlorine did not inhibit protein, DNA, or cell wall synthesis though it significantly inhibited RNA synthesis . Furthermore, using heterologous systems (mammalian ribosomes and C . albicans cytosolic factors) the data suggest that the inhibitory action of aspirochlorine is not exerted through a direct interaction with C . albicans EF-1 or EF-2.

J Nat Prod, 1999 May, 62(5), 767 - 9
Two auronols from Pseudolarix amabilis; Li XC et al.; Two new auronols, amaronols A (1) and B (2), were isolated from the bark of Pseudolarix amabilis, along with pseudolaric acid B (3), pseudolaric acid C (4), demethoxydeacetoxy-pseudolaric acid B (5), pseudolaric acid B-beta-D-glucoside (6), pseudolaric acid A-beta-D-glucoside (7), and myricetin (8) . The structures of amaronols A and B were established by spectral data interpretation as 2,4,6-trihydroxy-2-{(3',4',5'-trihydroxyphenyl) methyl}-3(2H)-benzofuranone and 2,4,6-trihydroxy-2-{(3', 5'-dihydroxy-4'-methoxyphenyl) methyl}-3(2H)-benzofuranone, respectively . Antimicrobial testing results of the eight compounds indicated that only pseudolaric acid B was active against Candida albicans (MIC, 3.125 microg/mL; MFC, 6.25 microg/mL), while myricetin was marginally active against Trichophyton mentagrophytes (MIC, 50 microg/mL).

J Nat Prod, 1999 May, 62(5), 678 - 80
Antifungal metabolites from the marine sponge Pachastrissa sp.: new bengamide and bengazole derivatives; Fernandez R et al.; This paper reports the studies of components of an undescribed sponge in the genus Pachastrissa sp., collected along the Djibouti coast . The extract showed activity against Candida albicans . Six new bengazoles (1-6) and a new bengamide, named bengamide L (16), in addition to the known bengazoles (7-11), bengamides A (12), B (13), E (14), and F (15), and a lactone (17) are described in this paper . All structures were determined on the basis of spectroscopic studies.

Electrophoresis, 1999 Apr-May, 20(4-5), 1001 - 10
Two-dimensional gel electrophoresis as analytical tool for identifying Candida albicans immunogenic proteins; Pitarch A et al.; This paper reports the usefulness of two-dimensional gel electrophoresis followed by Western blotting with sera from patients with systemic candidiasis in the identification of the major Candida albicans antigens . In order to have different patterns of protein expression and subcellular localization, three types of protein preparations were obtained: cytoplasmic extracts, protoplast lysates and proteins secreted by protoplasts regenerating their cell wall . These proteins were separated by high-resolution two-dimensional electrophoresis using an immobilized pH gradient . Western blotting with sera from patients with systemic candidiasis allowed the detection of more than 18 immunoreactive proteins . Some of these proteins had different isoforms . All sera reacted with at least three C . albicans proteins and the most reactive serum detected up to eleven proteins . Some of these antigens, e.g., enolase and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), have been identified on the 2-D map . The most reactive proteins were enolase and a 34 kDa protein in the acidic part of the gel (pI 4-4.4) that was only detected in regenerating protoplast-secreted proteins . The identification of all these antigens would be useful for the development of diagnostic strategies.

J Drug Target, 1999, 6(5), 361 - 72
Light and electron microscopic findings in a model of human cutaneous candidosis based on reconstructed human epidermis following the topical application of different econazole formulations; Schaller M et al.; The effects of two commercially available econazole formulations (econazole nitrate cream, econazole liposome gel) on uninfected reconstructed human epidermis and on a model of human cutaneous candidosis were investigated . The morphological alterations of the reconstructed epidermis after infection and treatment were analysed with light and electron microscopy . The most important Candida albicans-specific alterations of the recently established in vitro model of human cutaneous candidosis were scaling, hyperkeratosis, parakeratosis, dyskeratosis and spongiosis . A single application of the cream to the uninfected reconstructed epidermis caused more epidermal barrier damage and irritative toxic effects than the liposome gel . Treatment of the modelled human cutaneous candidosis with the cream also resulted in increased toxic effects, e.g., enhancement of scaling with invasion of Candida albicans blastospores into the stratum corneum and intracellular vacuoles . After application of the liposomal preparation invasion of Candida albicans in the stratum corneum could not be detected and toxic effects were reduced . Some of the Candida albicans-specific alterations such as hyperkeratosis, focal thickening of the stratum corneum, dyskeratosis and parakeratosis were completely eliminated . The liposomal formulation increased slightly the morphological alterations of the blastospores . Remnants of the cream formulation could be detected only very rarely in the stratum corneum or the blastospores . The liposomal preparation showed a strong affinity for the Candida albicans cells and the stratum corneum . Intact liposomes could even be observed in the intercellular spaces of the upper stratum corneum . As successful treatment depends on the ability to target the liposomal agent to the wanted site of action, this might be useful for more effective treatment of cutaneous candidosis.

Diagn Microbiol Infect Dis, 1999 May, 34(1), 19 - 25
Distribution of Candida albicans genotypes among family members; Mehta SK et al.; Thirty-three families (71 subjects) were screened for the presence of Candida albicans in mouthwash or stool specimens; 12 families (28 subjects) were culture-positive for this yeast . An enrichment procedure provided a twofold increase in the recovery of C . albicans from mouthwash specimens . Nine of the twelve culture-positive families had two positive members each, two families had three positive members each, and one family had four positive members . Genetic profiles were obtained by three methods: pulsed-field gel electrophoresis; restriction endonuclease analysis, and random amplification of polymorphic DNA analysis . DNA fingerprinting of C . albicans isolated from one body site three consecutive times revealed that each of the 12 families carried a distinct genotype . No two families shared the same strain, and two or more members of a family commonly shared the same strain . Intrafamily genotypic identity (i.e., each member within the family harbored the same strain) was demonstrated in six families . Genotypes of isolates from husband and wife differed from one another in five families . All three methods were satisfactory in determining genotypes; however, we concluded that restriction endonuclease analysis provided adequate resolving power.

Crit Rev Microbiol, 1999, 25(1), 1 - 17
Molecular mechanisms of chromosomal rearrangement in fungi; Fierro F et al.; Both sexual and asexual fungi undergo chromosomal rearrangements, which are the main cause of karyotype variability among the populations . Different recombination processes can produce chromosomal reorganizations, both during mitosis and meiosis, but other mechanisms operate to limit the extent of the rearrangements; some of these mechanisms, such as the RIP (repeat-induced point mutations) of Neurospora crassa, have been well established for sexual fungi . In laboratory strains, treatments such as mutation and transformation enhance the appearance of chromosomal rearrangements . Different DNA sequences present in fungal genomes are able to promote these reorganizations; some of these sequences are involved in well-regulated processes (e.g., site-specific recombination) but most of them act simply as substrates for recombination events leading to DNA rearrangements . In Penicillium chrysogenum we have found that short specific DNA sequences are involved in tandem reiterations leading to amplification of the cluster of the penicillin biosynthesis genes . In some cases, specific chromosomal rearrangements have been associated with particular phenotypes (as occurs in adaptive-like mutants of Candida albicans and Candida stellatoidea), and they may play a role in genetic variability for environmental adaptation.

Clin Exp Immunol, 1999 May, 116(2), 291 - 8
Effects of glycyrrhizin, an active component of licorice roots, on Candida albicans infection in thermally injured mice; Utsunomiya T et al.; Due to the generation of burn-associated CD8+ CD11b+ TCR gamma/delta+ type 2 T cells (burn-associated type 2 T cells), the susceptibility of thermally injured mice to infection with C . albicans has been shown to be increased by up to 50-fold when compared with normal mice . Glycyrrhizin (GR), an active component of licorice roots, reduced the susceptibility of thermally injured mice to C . albicans infection to levels observed in normal mice . Thermally injured mice inoculated with CD4+ T cells from GR-treated mice were also resistant to C . albicans infection . The following demonstrated that susceptibility to fungal infection was similar in thermally injured mice and normal mice inoculated with T6S cells (a clone of burn-associated type 2 T cells) . This susceptibility of T6S mice (normal mice inoculated with T6S cells) was reversible by (i) administration of GR, (ii) inoculation of CD4+ T cells from GR-treated mice, and (iii) injection of a mixture of MoAbs targeted against type 2 cytokines (IL-4 and IL-10) . After stimulation with anti-CD3 MoAb, splenic T cells from thermally injured and T6S mice, treated with GR or inoculated with CD4+ T cells from GR-treated mice, did not have type 2 cytokines in culture supernatants . They were present in splenic T cell cultures from thermally injured and T6S mice that were treated with saline or inoculated with naive T cells . These results suggest that GR, by inducing CD4+ T cells which suppress type 2 cytokines produced by burn-associated type 2 T cells, improves the resistance of thermally injured mice to C . albicans . An anti-type 2 T cell action of the CD4+ T cells derived from GR-treated mice was previously described.

Microbiol Immunol, 1999, 43(3), 235 - 40
Effective inhibition of Candida albicans growth by the combination of murine peritoneal neutrophils and activated macrophages; Tansho S et al.; The effect of leukocytes on the anti-Candida activity of neutrophils was examined . Murine neutrophils which were purified from casein-induced peritoneal cells inhibited the mycelial growth of Candida albicans . This anti-Candida activity of neutrophils was augmented by the addition of spleen cells prepared from mice pretreated with bacterial lipopolysaccharide 3 hr before, but not from non-treated mice . The population in the spleen cells, which enhanced the anti-Candida activity of neutrophils, was plastic-plate adherent, nylon-fiber columns adherent and anti-Mac-1 antigen-positive . These immunological profiles suggested that the enhancing cells are classified to splenic macrophages . Peritoneal-exudated macrophages from mice treated with lipopolysaccharide also augmented the anti-Candida activity of neutrophils . These results suggest that the anti-Candida activity of neutrophils may be upregulated by activated macrophages.

FEBS Lett, 1999 Apr 23, 449(2-3), 105 - 10
A critical comparison of the hemolytic and fungicidal activities of cationic antimicrobial peptides; Helmerhorst EJ et al.; The hemolytic and fungicidal activity of a number of cationic antimicrobial peptides was investigated . Histatins and magainins were inactive against human erythrocytes and Candida albicans cells in phosphate buffered saline, but displayed strong activity against both cell types when tested in 1 mM potassium phosphate buffer supplemented with 287 mM glucose . The HC50/IC50 ratio, indicative of the therapeutic index, was about 30 for all peptides tested . PGLa was most hemolytic (HC50 = 0.6 microM) and had the lowest therapeutic index (HC50/IC50 = 0.5) . Susceptibility to hemolysis was shown to increase with storage duration of the erythrocytes and also significant differences were found between blood collected from different individuals . In this report, a sensitive assay is proposed for the testing of the hemolytic activities of cationic peptides . This assay detects subtle differences between peptides and allows the comparison between the hemolytic and fungicidal potency of cationic peptides.

Glycobiology, 1999 Jun, 9(6), 533 - 7
Purification and biochemical characterization of two soluble alpha-mannosidases from Candida albicans; Vazquez-Reyna AB et al.; Two soluble alpha-mannosidases, E-I and E-II, were purified from C . albicans yeast cells by a three-step procedure consisting of size exclusion and ion exchange chromatographies in Sepharose CL6B and Mono Q columns, respectively, and preparative nondenaturing electrophoresis . E-I and E-II migrated as monomeric polypeptides of 54.3 and 93.3 kDa in SDS-PAGE, respectively . Some biochemical properties of purified enzymes were investigated by using 4-methylumbelliferyl-alpha-D-mannopyranoside and p-nitrophenyl-alpha-D-mannopyranoside as substrates . Hydrolysis of both substrates by either enzyme was optimum at pH 6.0 with 50 mM Mes-Tris buffer and at 42 degrees C . Apparent Kmvalues for hydrolysis of 4-methylumbelliferyl-alpha-D-mannopyranoside and p-nitrophenyl-alpha-D-mannopyranoside by E-I were 0.83 microM and 2 . 4 mM, respectively . Corresponding values for E-II were 0.25 microM and 1.86 mM . Swansonine and deoxymannojirimicin strongly inhibited the hydrolysis of 4-methylumbelliferyl-alpha-D-mannopyranoside by both enzymes . On the contrary, hydrolysis of p-nitrophenyl-alpha-D-mannopyranoside by E-I and E-II was slightly stimulated or not affected, respectively, by both inhibitors . E-I and E-II did not depend on metal ions although activity of the latter was slightly stimulated by Mn2+and Ca2+in the range of 0.5-2 mM . At the same concentrations, Mg2+was slightly inhibitory of both enzymes . Substrate specificity experiments revealed that both E-I and E-II preferentially cleaved alpha-1,6 and alpha-1,3 linkages, respectively.

J Basic Microbiol, 1999, 39(2), 97 - 101
Properties of adenosine deaminase from Candida albicans; Challa A et al.; Adenosine deaminase (ADA; adenosine aminohydrolase, E.C . 3.5.4.4), a purine catabolic enzyme, was studied in Candida albicans, an opportunistic yeast that causes diseases ranging from superficial infections to the deep systemic disease, candidiasis, in immunosuppressed humans . The fungus was grown as a yeast form in LEE's synthetic medium, pH 4.5, at room temperature for various growth periods . Adenosine deaminase (ADA) activity was determined from the cell free extract by measuring the change in absorbance 265 nm resulting from the deamination of adenosine . In yeast form, maximum growth and ADA activity were found at 72 and 24 hours, respectively, whereas in the mycelial form both the growth and ADA activity were maximum after 48 hours . Among the three media tested, tryptic soy broth supported maximum growth and enzyme production, compared to LEE synthetic medium or SABOURAUD dextrose broth . The enzyme was active over the pH range 4-8 and the optimum temperature for ADA activity was found to be 37 degrees C.

Clin Exp Allergy, 1999 Jun, 29(6), 824 - 31
Candida albicans mannan- and protein-induced humoral, cellular and cytokine responses in atopic dermatitis patients; Savolainen J et al.; BACKGROUND: The cytokine observed most often in atopic dermatitis (AD) is IL-4, but a role for IL-5 and IFN-gamma in the late and delayed phase reactions has been suggested . In AD with head, neck and shoulder distribution, hypersensitivity to saprophytic yeasts is an important pathogenetic factor . The yeast allergens include both the mannan polysaccharides and the proteins . Mannans are major cross-reacting allergens likely to be involved in the pathogenesis of AD . OBJECTIVE: To characterize the humoral, lymphoproliferative and cytokine (IL-2, 4, 5 and IFN-gamma) responses of peripheral blood mononuclear cells (PBMCs) induced by Candida albicans mannan and protein antigens in AD . METHODS: Fifteen AD patients and seven healthy controls were included . Ficoll-isolated PBMCs were stimulated by PHA and laboratory-generated mannan and protein extracts of C . albicans . Lymphocyte proliferation was measured and cytokine production was studied by ELISA . The antigen-specific IgG and IgE antibodies were analysed by ELISA and nitrocellulose RAST . RESULTS: In AD mannan (P < 0.005) and protein (P < 0.002), specific IgE levels were higher than in healthy controls . Both mannan and protein-specific lymphoproliferations (both: P < 0.02) were higher in AD than in healthy controls . Mannan, but not protein, induced long lasting IL-2 and IL-4 productions from 24 h lasting up to 66-96 h and IL-5 and IFN-gamma productions with elevated levels at 66 and 96 h . The mannan-induced IL-2 (P = 0.015) and IFN-gamma (P < 0.005) were increased in AD as compared with healthy controls . Significant correlations were seen between the protein-induced proliferation responses and both serum total IgE (r = 0.59, P < 0.01) and protein-specific IgE (r = 0.65, P < 0.005) . The mannan-induced IL-2 responses correlated with the specific IgE (r = 0.62, P < 0.01) and proliferation (r = 0.51, P < 0.02) and S-IgE level (r = 0.71, P < 0 . 002) . Mannan-induced IL-4 and IFN-gamma productions also correlated (r = 0.43, P < 0.05) . CONCLUSIONS: C . albicans mannan induced elevated IL-2 and IFN-gamma responses in AD patients . The correlations of the cytokine responses with mannan-induced IgE and proliferation responses suggest that C . albicans mannan induced TH1 type cytokine responses are involved in AD.

Rev Esp Quimioter, 1998 Dec, 11(4), 339 - 43
{In vitro activity of fluconazole against Candida albicans isolated from blood culture}; Peman J et al.; The in vitro activity of fluconazole against 65 Candida albicans isolated from blood culture in 1995, 1996 and 1997 was studied by macrodilution and disk diffusion methods . The MIC ranged from 0.03 to 64 microg/ml, with 93.6% of strains being inhibited with 1 microg/ml fluconazole; the mode MIC was 0.25 microg/ml . Using this method, only one strain was resistant and another was susceptible depending on the dose . By diffusion, eight strains were susceptible, 53 intermediate, and four resistant . The strains susceptible by dilution were also susceptible by diffusion, but the strains resistant by diffusion were not always resistant by dilution . We find it therefore useful to determine the MIC of fluconazole to the C . albicans resistant by diffusion.

Biotechnol Appl Biochem, 1999 Jun, 29 ( Pt 3), 223 - 7
Generation of a highly immunogenic recombinant enolase of the human opportunistic pathogen Candida albicans; Sandini S et al.; Enolase, a 46 kDa glycolytic enzyme, is an immunodominant antigen of Candida albicans, an important human opportunistic pathogen . The full-length coding sequence of C . albicans enolase gene was subcloned into the prokaryotic expression vector pDS56/RBSII,His6/E- under the control of an inducible promoter to produce a His6-tagged enolase . The recombinant protein was purified to homogeneity by one-step nickel-chelate affinity chromatography . It was recognized by a monoclonal antibody specific for C . albicans enolase, as well as by anti-enolase antibodies present in human sera (IgG) . The recombinant protein promptly elicited antibody in mice and detected immune responses in normal human subjects that were comparable to those generated by the native C . albicans enolase . Thus this new recombinant enolase constitutes a valuable reagent for studying the possible role of this protein in anti-Candida immune response.

J Clin Lab Immunol, 1996, 48(1), 1 - 15
Immunological cross reactivity between Candida albicans and human tissue; Vojdani A et al.; An old concept to account for autoimmunity is the existence of immunologic cross reactions, or shared determinants between an exogenous agent and self antigen . To study molecular mimicry between the Candida antigen and an autoantigen, sera from clinical specimens were screened, based on seronegativity or positivity for thyroid, ovary and adrenal antibodies . Compared with tissue antibody negative sera and sera from healthy controls, samples from positive tissue antibody subjects exhibited significantly higher levels of Candida IgG (P < 0.001) IgM (P < 0.001) and IgA (P < 0.01) antibodies . While Candida antibodies were elevated in 60% of tissue antibody positive samples, these antibodies were present in only 7.5% of tissue antibody negative subjects and in 10% of healthy controls . Since PAGE electrophoresis showed similar bands mobility in Candida and different tissues, these positive antibodies and rabbit anti Candida antibodies were reacted in immunodiffusion and Western Blot Assay against Candida and tissue antigens, simultaneously . The results of immunodiffusion showed a clear precipitation line against tissue antigens when rabbit anti Candida or human positive Candida serum was used . Similarly, Western Blot Assays with rabbit or human anti Candida serum showed several positive bands with Candida and one or two positive bands with different tissues . The common antigens were located in the regions of 72 and 36 KD . The 72 KD was detected in capsule antigens, placenta, ovary, adrenal, thymus, liver, pancreas, spleen, brain and kidney, but not in sperm or epithelial cell antigen . The 36 KD antigen was positive in placenta, spleen adrenal, pancreas and capsule tissues . Absorbtion of sera containing high levels of Candida antibodies with tissue antigens caused 10-15% reduction in antibody titers . Moreover, treatment of thyroid antibody positive sera with C . Albicans caused a similar reduction in thyroid antibody levels . These reductions in antibody levels are an additional support for cross reactivity between C . Albicans and mammalian tissues . A demonstration of immunological cross reactivity between Candida and human tissues may be associated with the possible pathogenic role of Candida Albicans in the development of autoimmune diseases which warrants further investigation.

J Chemother, 1999 Apr, 11(2), 131 - 6
Nosocomial Candida krusei fungemia in cancer patients: report of 10 cases and review; Krcmery V Jr et al.; The risk factors, therapy and outcome of ten cases of fungemia due to Candida krusei, appearing during the last 10 years in a single national cancer institution, are analyzed . Univariate analyses did not find any specific risk factors in comparison to 51 Candida albicans fungemias appearing at the same institution and with a similar antibiotic policy . Association with prior fluconazole prophylaxis was not confirmed because only one case appeared in a patient previously treated with fluconazole . However, attributable and crude mortality due to C . krusei fungemias was higher than for C . albicans fungemia . The authors review 172 C . krusei fungemias published within the last 10 years to compare with the incidence, therapy and outcome of C . krusei fungemia from our cancer institute.

Mol Cell Biol, 1999 Jun, 19(6), 4019 - 27
A G1 cyclin is necessary for maintenance of filamentous growth in Candida albicans; Loeb JD et al.; Candida albicans undergoes a dramatic morphological transition in response to various growth conditions . This ability to switch from a yeast form to a hyphal form is required for its pathogenicity . The intractability of Candida to traditional genetic approaches has hampered the study of the molecular mechanism governing this developmental switch . Our approach is to use the more genetically tractable yeast Saccharomyces cerevisiae to yield clues about the molecular control of filamentation for further studies in Candida . G1 cyclins Cln1 and Cln2 have been implicated in the control of morphogenesis in S . cerevisiae . We show that C . albicans CLN1 (CaCLN1) has the same cell cycle-specific expression pattern as CLN1 and CLN2 of S . cerevisiae . To investigate whether G1 cyclins are similarly involved in the regulation of cell morphogenesis during the yeast-to-hypha transition of C . albicans, we mutated CaCLN1 . Cacln1/Cacln1 cells were found to be slower than wild-type cells in cell cycle progression . The Cacln1/Cacln1 mutants were also defective in hyphal colony formation on several solid media . Furthermore, while mutant strains developed germ tubes under several hypha-inducing conditions, they were unable to maintain the hyphal growth mode in a synthetic hypha-inducing liquid medium and were deficient in the expression of hypha-specific genes in this medium . Our results suggest that CaCln1 may coordinately regulate hyphal development with signal transduction pathways in response to various environmental cues.

J Bacteriol, 1999 May, 181(10), 3058 - 68
Role of the mitogen-activated protein kinase Hog1p in morphogenesis and virulence of Candida albicans; Alonso-Monge R et al.; The relevance of the mitogen-activated protein (MAP) kinase Hog1p in Candida albicans was addressed through the characterization of C . albicans strains without a functional HOG1 gene . Analysis of the phenotype of hog1 mutants under osmostressing conditions revealed that this mutant displays a set of morphological alterations as the result of a failure to complete the final stages of cytokinesis, with parallel defects in the budding pattern . Even under permissive conditions, hog1 mutants displayed a different susceptibility to some compounds such as nikkomycin Z or Congo red, which interfere with cell wall functionality . In addition, the hog1 mutant displayed a colony morphology different from that of the wild-type strain on some media which promote morphological transitions in C . albicans . We show that C . albicans hog1 mutants are derepressed in the serum-induced hyphal formation and, consistently with this behavior, that HOG1 overexpression in Saccharomyces cerevisiae represses the pseudodimorphic transition . Most interestingly, deletion of HOG1 resulted in a drastic increase in the mean survival time of systemically infected mice, supporting a role for this MAP kinase pathway in virulence of pathogenic fungi . This finding has potential implications in antifungal therapy.

Mol Microbiol, 1999 May, 32(3), 547 - 56
Sequential gene disruption in Candida albicans by FLP-mediated site-specific recombination; Morschhauser J et al.; The genetic manipulation of the human fungal pathogen Candida albicans is difficult because of its diploid genome, the lack of a known sexual phase and its unusual codon usage . We devised a new method for sequential gene disruption in C . albicans that is based on the repeated use of the URA3 marker for selection of transformants and its subsequent deletion by FLP-mediated, site-specific recombination . A cassette was constructed that, in addition to the URA3 selection marker, contained an inducible SAP2P-FLP fusion and was flanked by direct repeats of the minimal FLP recognition site (FRT) . This URA3 flipper cassette was used to generate homozygous C . albicans mutants disrupted for both alleles of either the CDR4 gene, encoding an ABC transporter, or the MDR1 gene, encoding a membrane transport protein of the major facilitator superfamily . After insertion of the URA3 flipper into the first copy of the target gene, the whole cassette could be efficiently excised by induced FLP-mediated recombination, leaving one FRT site in the disrupted allele of the target gene . The URA3 flipper was then used for another round of mutagenesis to disrupt the second allele . Deletion of the cassette from primary and secondary transformants occurred exclusively by intrachromosomal recombination of the FRT sites flanking the URA3 flipper, whereas interchromosomal recombination between FRT sites on the homologous chromosomes was never observed . This new gene disruption strategy facilitates the generation of specific, homozygous C . albicans mutants as it eliminates the need for a negative selection scheme for marker deletion and minimizes the risk of mitotic recombination in sequential disruption experiments.

Mol Microbiol, 1999 May, 32(3), 533 - 46
Host-induced, stage-specific virulence gene activation in Candida albicans during infection; Staib P et al.; An understanding of the complex interactions between pathogenic microbes and their host must include the identification of gene expression patterns during infection . To detect the activation of virulence genes in the opportunistic fungal pathogen Candida albicans in vivo by host signals, we devised a reporter system that is based on FLP-mediated genetic recombination . The FLP gene, encoding the site-specific recombinase FLP, was genetically modified for expression in C . albicans and fused to the promoter of the SAP2 gene that codes for one of the secreted aspartic proteinases, which are putative virulence factors of C . albicans . The SAP2P-FLP fusion was integrated into one of the SAP2 alleles in a strain that contained a deletable marker that conferred resistance to mycophenolic acid and was flanked by direct repeats of the FLP recognition target (FRT) . Using this reporter system, a transient gene induction could be monitored at the level of single cells by the mycophenolic acid-sensitive phenotype of the colonies generated from such cells after FLP-mediated marker excision . In two mouse models of disseminated candidiasis, SAP2 expression was not observed in the initial phase of infection, but the SAP2 gene was strongly induced after dissemination into deep organs . In contrast, in a mouse model of oesophageal candidiasis in which dissemination into internal organs did not occur, no SAP2 expression was detected at any time . Our results support a role of the SAP2 gene in the late stages of an infection, after fungal spread into deep tissue . This new in vivo expression technology (IVET) for a human fungal pathogen allows the detection of virulence gene induction at different stages of an infection, and therefore provides clues about the role of these genes in the disease process.

Immunopharmacol Immunotoxicol, 1999 May, 21(2), 331 - 42
Protection of immunosuppressed mice from lethal Candida infection by oral administration of a kampo medicine, hochu-ekki-to; Abe S et al.; The protective effect of a Kampo medicine, Hochu-ekki-to (TJ-41) on experimental candidiasis in immunosuppressed mice was investigated . ICR mice were immunosuppressed by injection of prednisolone or cyclophosphamide, given TJ-41 orally and challenged intravenously with Candida albicans (day 0) . Treatments with a daily dose of 1 g/kg/day of TJ-41 for 8 days from day-4 or for 4 days from day 0 significantly prolonged the life span of the Candida-infected mice pretreated with prednisolone . The latter treatment appeared to inhibit the colonization of Candida in kidneys of the infected mice . These results suggest that Hochu-ekki-to can be used as a therapeutic agent against candidiasis in patients with glucocorticoid-induced immunosuppression.

Yeast, 1999 Apr, 15(6), 507 - 11
Heterologous URA3MX cassettes for gene replacement in Saccharomyces cerevisiae; Goldstein AL et al.; Heterologous gene replacement cassettes are powerful tools for dissecting gene function in Saccharomyces cerevisiae . Their primary advantages over homologous gene replacement cassettes include reduced gene conversion (leading to efficient site-specific integration of the cassette) and greater independence of strain background . Perhaps the most widely used cassettes are the MX cassettes containing the dominant selectable kanamycin resistance gene (kanr), which confers resistance to G418 (Wach et al., 1994) . One limitation of the kanMX cassettes is that they are not counterselectable and therefore not readily recyclable, which is important when constructing strains with more than one gene deletion . To address this limitation, and to expand the choices of heterologous markers, we have created two new MX cassettes by replacing the kanr ORF from plasmids pFA6-kanMX3 and pFA6-kanMX4 with the Candida albicans URA3 ORF . These plasmids, pAG60 (CaURA3MX4) and pAG61 (CaURA3MX3) are identical to the kanMX cassettes in all other respects but have the added advantage of being counterselectable and therefore readily recyclable in S . cerevisiae.

Drug Discov Today, 1999 Jan, 4(1), 17 - 26
Computer-aided target selection-prioritizing targets for antifungal drug discovery; Spaltmann F et al.; The entire DNA sequence of the Saccharomyces cerevisiae genome was completed in 1996 and represents the first entirely decoded eukaryotic genome . Because major human pathogenic fungi such as Candida albicans are closely related to S . cerevisiae on a molecular level, the question arises as to how this new information can be used to identify and prioritize those genes that are most suitable as targets for antimycotic drug discovery . To tackle this challenge, a software tool called CATS (computer-aided target selection) was developed . The authors describe how it allows an automated and periodically updated assessment of all S . cerevisiae genes to be carried out with regard to their suitability as antifungal targets.

Nippon Ishinkin Gakkai Zasshi, 1999, 40(2), 79 - 83
{Fungi and atopic dermatitis}; Hiruma M et al.; Attention has recently been centered on fungi as aggravating factors of atopic dermatitis (AD) due to the frequent detection of IgE antibodies to fungi in patients with severe AD and to positive response of some cases of AD to antifungal therapy . Malassezia sp.: In AD patients with prominent symptoms in the head and neck, areas prone to colonization by Malassezia, the titers of specific anti-Malassezia IgE antibodies are high, which positively correlate with the total IgE value and the severity of AD . The patch test against Malassezia antigens is positive . The rate of isolation of Malassezia from the skin of AD patients is higher than that from the skin of healthy control subjects . Candida sp.: In patients with severe AD, the rate of positive skin prick tests for Candida is high, and a correlation exists between positive skin prick test results and the presence of Candida albicans in nasopharynx . However, the reactivity to Candida antigens in the patch tests is reduced, and a negative correlation is seen . There is no difference between the isolation rate of C . albicans from patients with adult-type AD and normal controls . However, AD patients give a significantly greater number of separate colonies . The range of efficacy rate of antifungal therapy of AD is reported to be 50-65 % . The efficacy rate of our own trial falls within this range . Following treatment, the rate of isolation of fungi decrease significantly, and the titers of specific antifungal IgE antibodies are not statistically significant . The clearance of fungi from the tissue following antifungal therapy probably results in the suppression of direct or indirect inflammatory reaction caused by the fungi . We therefore consider antifungal therapy as one of the second-line therapies to be administered in AD cases resistant to conventional basic therapy.

Arch Pharm Res, 1999 Apr, 22(2), 143 - 50
Phototoxicity of melatonin; Kim YO et al.; Melatonin (MLT), N-acetyl-5-methoxytryptamine, is mainly secreted by the pineal gland . The ultraviolet (UV), infrared (IR) and 1H-NMR spectra of irradiated and non-irradiated MLT were measured, and phototoxicity tests of MLT, anthracene (positive control) and sodium lauryl sulfate (SLS, negative control) were performed . The methods employed include both in vitro tests such as MTS assay using the human fibroblast cell and yeast growth inhibition assay using Candida albicans and in vivo method using the skin of guinea pig . UV absorption spectra and 1H-NMR spectra of MLT were changed by UVA (365 nm, 15 J/cm2), but IR spectra of MLT were not changed . The fifty percent inhibitor concentration (IC50) ratio (UV-/UV+) of MLT was 10 . The inhibition zone of irradiated-paper disks treated with MLT was not observed . According to the results of histopathological examination, no pathologic lesion was observed in the non-irradiated group, but slight degeneration of keratinocytes in the epidermis, hemorrhage and vasodilation in dermis were observed in the irradiated group . These results indicate that the molecular structure of MLT is altered by UVA to unidentified photoproducts and a moderate phototoxicity of MLT is predicted.

J Infect Dis, 1999 Jun, 179(6), 1477 - 84
Candida albicans mannan extract-protein conjugates induce a protective immune response against experimental candidiasis; Han Y et al.; Candida albicans mannan extracts encapsulated in liposomes were previously used to stimulate mice to produce antibodies protective against candidiasis . In the present study, mannan-protein conjugates without liposomes were tested as vaccine candidates . Mannan extracts were coupled to bovine serum albumin, and isolated conjugates consisted of carbohydrate and protein at a ratio of 0.7-1.0 . Vaccination of mice with the conjugate and an adjuvant yielded antiserum that contained Candida agglutinins . Vaccinated mice challenged with yeast cells had a mean survival time of 56 days, compared with <13 days for control groups . The antiserum protected naive animals against disseminated disease . Naive mice given the antiserum intravaginally developed 79% fewer fungal colony-forming units, compared with control groups . The serum-protective factor was stable at 56 degrees C and was removed by adsorption with yeast cells . It is concluded that the conjugate vaccine can induce protective antibody responses against experimental disseminated candidiasis and Candida vaginal infection.

EMBO J, 1999 May 4, 18(9), 2580 - 92
Phenotypic switching in Candida albicans is controlled by a SIR2 gene; Perez-Martin J et al.; We report the cloning of a gene from the human fungal pathogen Candida albicans with sequence and functional similarity to the Saccharomyces cerevisiae SIR2 gene . Deletion of the gene in C . albicans produces a dramatic phenotype: variant colony morphologies arise at frequencies as high as 1 in 10 . The morphologies resemble those described previously as part of a phenotypic switching system proposed to contribute to pathogenesis . Deletion of SIR2 also produces a high frequency of karyotypic changes . These and other results are consistent with a model whereby Sir2 controls phenotypic switching and chromosome stability in C.albicans by organizing chromatin structure.

FEMS Microbiol Lett, 1999 Apr 15, 173(2), 475 - 81
Membrane fluidity affects functions of Cdr1p, a multidrug ABC transporter of Candida albicans; Smriti et al.; Earlier, we have shown that the overexpression of an ABC transporter, CDR1, is involved in the emergence of multidrug resistance in Candida albicans . In this study, we checked its function in vivo by expressing it in different isogenic Saccharomyces cerevisiae erg mutants, which accumulated various intermediates of the ergosterol biosynthesis and thus altered the membrane fluidity . Functions like the accumulation of rhodamine 123, beta-estradiol, fluconazole and floppase activity associated with Cdr1p were measured to ascertain their responses to an altered membrane phase . The floppase activity appeared to be favoured by an enhanced membrane fluidity, while the effluxing of substrates and Cdr1p's ability to confer multidrug resistance were significantly reduced . We demonstrate that only some of the functions of Cdr1p were affected by an altered lipid environment.

Infect Immun, 1999 May, 67(5), 2482 - 90
In vivo analysis of secreted aspartyl proteinase expression in human oral candidiasis; Naglik JR et al.; Secreted aspartyl proteinases are putative virulence factors in Candida infections . Candida albicans possesses at least nine members of a SAP gene family, all of which have been sequenced . Although the expression of the SAP genes has been extensively characterized under laboratory growth conditions, no studies have analyzed in detail the in vivo expression of these proteinases in human oral colonization and infection . We have developed a reliable and sensitive procedure to detect C . albicans mRNA from whole saliva of patients with oral C . albicans infection and those with asymptomatic Candida carriage . The reverse transcription-PCR protocol was used to determine which of the SAP1 to SAP7 genes are expressed by C . albicans during colonization and infection of the oral cavity . SAP2 and the SAP4 to SAP6 subfamily were the predominant proteinase genes expressed in the oral cavities of both Candida carriers and patients with oral candidiasis; SAP4, SAP5, or SAP6 mRNA was detected in all subjects . SAP1 and SAP3 transcripts were observed only in patients with oral candidiasis . SAP7 mRNA expression, which has never been demonstrated under laboratory conditions, was detected in several of the patient samples . All seven SAP genes were simultaneously expressed in some patients with oral candidiasis . This is the first detailed study showing that the SAP gene family is expressed by C . albicans during colonization and infection in humans and that C . albicans infection is associated with the differential expression of individual SAP genes which may be involved in the pathogenesis of oral candidiasis.

J Physiol Biochem, 1998 Dec, 54(4), 203 - 15
Opioid peptides and immunodysfunction in patients with major depression and anxiety disorders; Castilla-Cortazar I et al.; To assess cell-mediated immunity in depression and anxiety disorders and to elucidate whether immunodysfunction might be related to a high opioid activity, a prospective study of patients with major depression (n = 34) or anxiety disorders (n = 21) was performed . Cellular immunity tests, the in vitro effects of naloxone on monocytes, and beta-endorphin plasma levels were investigated . Peripheral blood mononuclear cells and some monocyte parameters were determined by flow cytometry . Natural killer (NK) cell activity was studied by cytotoxicity, gamma-interferon production by a standard bioassay, monocytic phagocytosis by ingestion of Candida albicans and latex, and blastogenesis by stimulation with phytohaemaglutinin . In major depression and anxiety: 1) a marked reduction in the number of monocytes that ingested particles and expressed cytoskeletal intermediate filaments and surface structures (CR1 receptors and HLA-DR antigens); 2) a monocytosis that was not able to normalize the count of functioning monocytes; 3) an in vitro correction of the monocyte dysfunction by naloxone; 4) a decrease in NK cell number and activity; and 6) an anergy to candidin and tuberculin and a diminished lectin-induced blastogenesis were observed . Some of these immune changes correlated closely with plasma beta-endorphin abnormally high in all the cases . In conclusion, a naloxone-reversible monocyte dysfunction, associated to decreased NK activity and cell-mediated hypersensitivity, was found together with high of beta-endorphin plasma levels . In addition, results suggest that these immunological alterations may be useful in the clinical management of patients with these psychiatric diseases.

FEMS Immunol Med Microbiol, 1999 Apr, 23(4), 343 - 54
Characterization of Candida albicans antigenic determinants by two-dimensional polyacrylamide gel electrophoresis and enhanced chemiluminescence; Barea PL et al.; The use of a two-dimensional polyacrylamide gel electrophoresis joined with Western blotting allowed us to investigate the reactivities of antibodies present in sera from mice and humans to antigens of Candida albicans blastoconidia . The analysis of the antibody response in the two models studied and the comparison between the antibody response in infected and noninfected individuals showed that the infection by C . albicans produces changes in the antibody response which may be of relevance in the serodiagnosis of invasive candidiasis . These changes include the induction of antibodies against new antigens, the disappearance of antibodies against a group of antigens and variations in the reactivity of antibodies directed to a different group of antigens . The technique used resolved the isoforms of several antigens including enolase . It is concluded that the antibody response in humans and mice with candidiasis is not homogeneously directed to all the isoforms of an antigen.

FEMS Immunol Med Microbiol, 1999 Apr, 23(4), 289 - 93
An investigation into the antimicrobial effects of adrenomedullin on members of the skin, oral, respiratory tract and gut microflora; Allaker RP et al.; Adrenomedullin, a novel vasoactive peptide, is known to be expressed by many surface epithelial cells and it was postulated that this peptide may have a protective role . The objective of the study was to assess the antimicrobial activity of adrenomedullin against members of the human skin, oral, respiratory tract and gut microflora using disc diffusion and broth microdilution assays . All strains of bacteria screened in an agar diffusion assay were sensitive; gram-positive and gram-negative bacteria were equally susceptible . No activity against the yeast Candida albicans was observed . In a broth microdilution assay, minimum inhibitory and minimum bacteriocidal concentrations ranged from 7.75 x 10(-1) to 12.5 microg ml(-1) and 0.003 to > 25.0 microg ml(-1), respectively . We propose an antimicrobial role for adrenomedullin . participating in the prevention of local infection, thus contributing to host defence systems.

FASEB J, 1999 May, 13(8), 823 - 32
Disruption of filamentous actin inhibits human macrophage fusion; DeFife KM et al.; The foreign body reaction to implanted biomaterials, characterized by the presence of macrophages and foreign body giant cells (FBGC), can result in structural and functional failure of the implant . Recently, we have shown that interleukin-4 and interleukin-13 can independently induce human macrophage fusion to form FBGC via a macrophage mannose receptor (MR) -mediated pathway . The MR is believed to mediate both endocytosis of glycoproteins and phagocytosis of microorganisms, which bear terminal mannose, fucose, N-acetylglucosamine, or glucose residues . Polarization of microfilaments to closely apposed macrophage membranes as observed with fluorescence confocal microscopy led us to ask whether MR-mediated fusion occurred via a filamentous actin-dependent pathway . Cytochalasins B and D and latrunculin-A, agents that disrupt microfilaments, inhibited macrophage fusion in a concentration-dependent manner . The concentrations of cytochalasins D and B that inhibited fusion did not significantly decrease macrophage adhesion, spreading, or motility but did inhibit internalization of Candida albicans during interleukin-13-enhanced, MR-mediated phagocytosis . Very low concentrations of cytochalasin B (< 2 microM) induced a slight enhancement of macrophage fusion . Taken together, the results of this study suggest that cytokine-induced, MR-mediated macrophage fusion requires an intact F-actin cytoskeleton and that the mechanism of fusion is similar to phagocytosis.--DeFife, K . M., Jenney, C . R., Colton, E., Anderson, J . M . Disruption of filamentous actin inhibits human macrophage fusion.

Antimicrob Agents Chemother, 1999 May, 43(5), 1163 - 9
Formation of azole-resistant Candida albicans by mutation of sterol 14-demethylase P450; Asai K et al.; The sterol 14-demethylase P450 (CYP51) of a fluconazole-resistant isolate of Candida albicans, DUMC136, showed reduced susceptibility to this azole but with little change in its catalytic activity . Twelve nucleotide substitutions, resulting in four amino acid changes, were identified in the DUMC136 CYP51 gene in comparison with a reported CYP51 sequence from a wild-type, fluconazole-susceptible C . albicans strain . Seven of these substitutions, including all of those causing amino acid changes, were located within a region covering one of the putative substrate recognition sites of the enzyme (SRS-1) . Polymorphisms within this region were observed in several C . albicans isolates, and some were found to be CYP51 heterozygotes . Among the amino acid changes occurring in this region, only an alteration of Y132 was common among these fluconazole-resistant isolates, which suggests the importance of this residue to the fluconazole resistance of the target enzyme . DUMC136 and another fluconazole-resistant isolate were homozygotes with respect to CYP51, although the typical wild-type, fluconazole-susceptible C . albicans was a CYP51 heterozygote . These findings suggest that part of the fluconazole-resistant phenotype of C . albicans DUMC136 was acquired through a mutation-prone area of CYP51, an area which might promote the formation of fluconazole-resistant CYP51, along with a mechanism(s) which allows the formation of a homozygote of this altered CYP51 in this diploid pathogenic yeast.

Antimicrob Agents Chemother, 1999 May, 43(5), 1034 - 41
Assessment of the effect of amphotericin B on the vitality of Candida albicans; Liao RS et al.; The processes involved in cell death are complex, and individual techniques measure specific fractions of the total population . The interaction of Candida albicans with amphotericin B was measured with fluorescent probes with different cellular affinities . These were used to provide qualitative and quantitative information of physiological parameters which contribute to fungal cell viability . SYBR Green I and 5,(6)-carboxyfluorescein were used to assess membrane integrity, and bis-(1,3-dibutylbarbituric acid)trimethine oxonol and 3,3-dihexyloxacarbocyanine iodide were used to evaluate alterations in membrane potential . The fluorescent indicators were compared with replication competency, the conventional indicator of viability . By using these tools, the evaluation of the response of C . albicans to amphotericin B time-kill curves delineated four categories which may represent a continuum between alive and dead . The data showed that replication competency (CFU per milliliter) as determined by conventional antifungal susceptibility techniques provided only an estimate of inhibition . Interpretation of fluorescent staining characteristics indicated that C . albicans cells which were replication incompetent after exposure to greater than 0.5 microgram of amphotericin B per ml still maintained degrees of physiological function.

J Antimicrob Chemother, 1999 Mar, 43(3), 419 - 22
Non-albicans oral candidosis in HIV-positive patients; Cartledge JD et al.; Specimens from HIV-positive patients with oral candidosis were taken for culture, species identification and azole susceptibility testing, which was correlated with treatment outcome . Of 921 specimens, 95 yielded non-albicans species, mainly from patients with low CD4 lymphocyte counts and extensive previous azole exposure . Most non-albicans isolates were from specimens co-infected with Candida albicans, complicating the interpretation of in-vitro susceptibility results, which accurately predicted antifungal failure when the non-albicans species was isolated alone . Eighty-five non-albicans isolates were resistant to fluconazole in vitro . Of 149 courses of azole therapy prescribed, 115 failed to clear non-albicans candidosis clinically . Culture media that discoloured in the presence of non-albicans colonies might, therefore, guide therapy.

J Antimicrob Chemother, 1999 Mar, 43(3), 411 - 3
Comparisons of the effects of fungicidal and fungistatic antifungal agents on the morphogenetic transformation of Candida albicans; Hawser S et al.; Eleven different antifungal agents were compared, and their ability to inhibit the morphogenetic transformation of Candida albicans was examined together with their ability to inhibit growth, as measured by MIC methodology . The fungicidal potential of each agent was also determined . Of the antifungal agents tested, only amphotericin B, mulundocandin and aculeacin inhibited the transformation at sub-MIC values; all three agents showed fungicidal activity at concentrations close to the MIC . All other agents were fungicidal only at concentrations much higher than the MIC and inhibited the morphogenetic transformation only at concentrations above the MIC . These data suggest that fungicidal antifungal agents are more likely to act by inhibiting the morphogenetic transformation of C . albicans while fungistatic agents are unable to do so and are more likely to block growth by budding.

Jpn J Antibiot, 1999 Feb, 52(2), 146 - 52
{The in vitro properties of a new hydroxypyridone antimycotic rilopirox, with special reference to its anti-Candida activity}; Harada I et al.; In vitro properties of a new hydroxypyridone antimycotic rilopirox (RIL) with special reference to its anti-Candida activities were studied in comparison with the three reference drugs, ciclopirox olamine (CPO), oxiconazole nitrate (OCZ) and isoconazole nitrate (ICZ), using several strains of Candida albicans and Candida glabrata as the test organisms . RIL was potently fungicidal for growing cultures of these Candida strains, whereas all the three reference drugs were slightly fungicidal or fungistatic . Unlike OCZ and ICZ whose anti-Candida activity was decreased by lowering pH or adding serum to culture media, the activity of RIL was scarecely affected by change in pH or serum addition . However, RIL became less potent in the presence of Fe3+ at concentrations of 10(-5) mmol/ml or above . These findings suggest that RIL will be useful as a topical anti-Candida agent.

Med Pediatr Oncol, 1999 May, 32(5), 344 - 8
Fungal colonization and infection in children with acute leukemia and lymphoma during induction therapy; Gozdasoglu S et al.; BACKGROUND: Fungal infection represents a growing problem in children with hematologic malignancies . During chemotherapy induced neutropenia, colonization with fungi is considered a major risk factor for subsequent fungal infection . The rates and risk factors for mycotic infections in pediatric oncology patients is undetermined, particularly for centers in developing countries . The aim of this study was to evaluate the rates and risk factors of fungal colonization in children with acute leukemia and lymphoma at one of the major pediatric hematology/oncology centers in Turkey . PROCEDURE: Fifty-two consecutive children newly diagnosed with acute leukemia and lymphoma during intensive remission induction therapy were evaluated for the occurrence of fungal colonization (defined as at least one positive surveillance culture) and infection . RESULTS: Thirty-six of the 52 patients (69.2%) were colonized by Candida albicans which was the only fungus isolated from surveillance cultures . There were three (5.8%) proven systemic fungal infections: two cases of candidemia and one case of brain abscess with Aspergillus spp . isolated from tissue . All patients with fungal colonization were receiving prophylactic or curative antibiotics . No significant association was found between type of disease and fungal colonization, but there was a significant association with neutropenia . CONCLUSIONS: Our findings suggest that there is a high rate of fungal colonization in children receiving remission induction therapy for acute leukemia and lymphoma . Limiting the use of antibiotics and instituting antifungal chemoprophylaxis may decrease the rate, while the early initiation of empiric antifungal therapy in patients with fever and suspected mycotic colonization may increase survival in these patients.

FEMS Immunol Med Microbiol, 1999 Mar, 23(3), 229 - 34
Clinical strains of Candida albicans express the surface antigen glyceraldehyde 3-phosphate dehydrogenase in vitro and in infected tissues; Gil ML et al.; We have previously described the presence of an enzymatically active form of glyceraldehyde 3-phosphate-dehydrogenase (GAPDH) in the cell surface of Candida albicans ATCC 26555 which is also a fibronectin and laminin binding protein . Immunohistochemical analysis of tissue sections from patients with disseminated candidiasis with a polyclonal antiserum to GAPDH from C . albicans (PAb anti-CA-GAPDH) revealed that the enzyme is expressed at the surface of fungal cells in infected tissues . The same PAb detected the presence of GAPDH species, with a molecular mass of approximately 33 kDa, in cell wall extracts obtained from clinical isolates of the fungus . These cell surface-bound GAPDH moieties exhibited a dose-dependent dehydrogenase activity . These results indicate that this cell surface-bound GAPDH plays a role during infection probably contributing to the attachment of fungal cells to host tissues.

Microbiology, 1999 Mar, 145 ( Pt 3), 695 - 701
Monoclonal antibody 3H8: a useful tool in the diagnosis of candidiasis; Marcilla A et al.; In a previous series of experiments six mAbs were obtained against cell wall extracts of Candida albicans ATCC 26555 . After several studies only one of them, designated 3H8, has been used to produce a commercial kit for the rapid diagnosis of candidiasis, Bichro-latex albicans (Fomouze Diagnostics) . The present study involved the generation and characterization of this mAb as an immunoglobulin G1 which recognizes mannoproteins of high molecular mass present in the C . albicans cell wall . ELISA assays showed that the presence of the epitope recognized by mAb 3H8 was similar in both yeast and mycelial cell walls of C . albicans, in contrast to the epitope for mAb 1B12, which is mainly expressed in the yeast cell wall . The 3H8 epitope was located at the external surface in C . albicans ATCC 26555, whereas it is partially cryptic in the cell wall in other C . albicans strains . No reaction was observed with other Candida species . Immunohistochemical studies using this antibody demonstrated that it specifically recognized C . albicans in tissue, detecting mycelial forms and, to a lesser extent, blastospores, suggesting that it is also a valuable tool in the evaluation of fungal infections in paraffin-embedded tissue, particularly when identification is required.

Microbiology, 1999 Mar, 145 ( Pt 3), 689 - 94
Characterization of a haemolytic factor from Candida albicans; Watanabe T et al.; The culture supernatant of Candida albicans promoted the disruption of human red blood cells (RBCs) . The haemolytic activity was detected in a sugar-rich fraction (about 200 kDa) from Sephacryl S-100 chromatography . As the haemolytic activity was adsorbed by concanavalin A-Sepharose, the haemolytic factor may be a mannoprotein . The activity was inactivated by periodate oxidation, indicating that the sugar moiety of the mannoprotein played an important role in the haemolysis . The structure of the sugar moiety of the mannoprotein was identified as a cell-wall mannan by 1H-NMR analysis, and purified C . albicans mannan promoted the disruption of RBCs . The binding of mannan to RBCs was demonstrated by flow cytometric analysis and was inhibited by the addition of band 3 protein inhibitor, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) . The haemolysis caused by mannan was inhibited by DIDS, SITS (4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid) and bis(sulfosuccinimidyl) suberate, but not by pyridoxal 5-phosphate . These results indicated that a mannoprotein released from C . albicans bound to the band 3 protein on RBCs, thereby promoting their disruption.

Biochim Biophys Acta, 1999 Apr 19, 1427(2), 245 - 55
Copper- and zinc-containing superoxide dismutase and its gene from Candida albicans; Hwang CS et al.; Cytosolic copper- and zinc-containing superoxide dismutase was purified 136-fold with an overall yield of 2.5% to apparent electrophoretic homogeneity from the dimorphic pathogenic fungus, Candida albicans . The molecular mass of the native enzyme was 39.4 kDa and the enzyme was composed of two identical subunits with a molecular mass of 19.6 kDa . The enzyme was stable in the range of pH 4.0-9.0 and up to 55 degrees C . The ultraviolet-visible absorption spectrum of the enzyme showed the absorption band of copper- and zinc-containing superoxide dismutase at 660 nm . The atomic absorption analysis revealed that the enzyme contained 0.87 g-atom of copper and 0.79 g-atom of zinc per mole of subunit . The N-terminal amino acid sequence alignments up to the 40th residue showed that copper- and zinc-containing superoxide dismutase from C . albicans has high similarity to other eukaryotic copper- and zinc-containing superoxide dismutases . The sod1 encoding copper- and zinc-containing superoxide dismutase has been cloned using a polymerase chain reaction fragment as a probe . Sequence analysis of the sod1 predicted a copper- and zinc-containing superoxide dismutase that contains 154 amino acids with a molecular mass of 16143 Da and displayed 79%, 69%, and 57% sequence identity to the homologues of Neurospora crassa, Saccharomyces cerevisiae, and bovine, respectively . The cloned sod1 contained an intron of 245 nucleotides, which was verified by reverse transcription-polymerase chain reaction.

Retina, 1999, 19(2), 122 - 6
In vitro antimicrobial activity of silicone oil against endophthalmitis-causing agents; Ozdamar A et al.; PURPOSE: To investigate the antimicrobial activity of silicone oil against endophthalmitis-causing agents in vitro . METHODS: The antimicrobial activity of silicone oil was tested on Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Candida albicans, and Aspergillus spp . The bacteria and fungi were separately inoculated into 1,300 centistokes silicone oil . Control inoculations were done in two different media: physiologic saline and brain-heart infusion (BHI) for bacteria and Sabouraud broth and physiologic saline for fungi . From each medium, 0.001-mL samples were taken and plated in Petri dishes . After overnight incubation, colony-forming units (CFUs) were enumerated . Culturing from the initially prepared specimens, incubating overnight, and counting CFUs was repeated until no growth of microorganisms was seen in the silicone oil-containing media . Macroscopic photography of the colonies and light microscopic photography of microorganisms were performed . RESULTS: All the microorganisms showed an apparent decrease in CFUs, with elimination between 7 and 21 days in silicone oil . Colony-forming units of microorganisms remained stable in physiologic saline during the study, with the exception of gradual decrease in CFUs of S . aureus and S . epidermidis from the beginning of the third day . In BHI and Sabouraud broth, both bacteria and fungi showed a growth pattern that was compatible with the growth curve of microorganisms . CONCLUSION: Silicone oil has an antimicrobial activity against S . aureus, S . epidermidis, P . aeruginosa, C . albicans, and Aspergillus spp., which are common endophthalmitis-causing agents.

Diagn Microbiol Infect Dis, 1999 Apr, 33(4), 231 - 9
Evaluation of DNA-based typing procedures for strain categorization of Candida spp; Espinel-Ingroff A et al.; DNA-based procedures have replaced earlier epidemiologic methodologies that relied on nonreproducible and insensitive measurements of phenotypic characteristics to identify a specific strain as the source of infection . The reliability (interlaboratory percent agreement for strain delineation) and sensitivity (recognition of subtle strain-to-strain variation) of similar DNA-based typing systems by different laboratories were evaluated . Ten isolates (five epidemiologic-related and five unrelated strains each) of Candida albicans, C . lusitaniae, C . parapsilosis, C . tropicalis, and Candida (Torulopsis) glabrata were characterized in a blinded fashion by three laboratories . All 50 isolates were subtyped in each laboratory by electrophoretic karyotyping (EK) analysis using contour-clamped homogenous electric field (CHEF) electrophoresis protocols . In addition, two laboratories also performed restriction endonuclease analysis of genomic DNA (REAG) using the restriction endonucleases SfiI and BssHII followed by CHEF electrophoresis separation of resulting fragments . DNA strain identification of the 50 isolates by the three different laboratories using similar CHEF methodologies demonstrated the following species-dependent, interlaboratory reproducibility: C . tropicalis (82%), C . parapsilosis (83%), C . albicans (90%), C . lusitaniae (93%), and C . glabrata (100%) . In addition, agreement was higher by the CHEF method (83 to 100%), when compared with the strain types identified by the REAG (60 to 100%) method . Five to seven strains of each Candida species evaluated were detected by the different methodologies used for this study . This study indicates that these procedures are relatively discriminatory and reliable tools to study strain-to-strain variations in epidemiologic evaluations of these yeasts.

Chem Pharm Bull (Tokyo), 1999 Mar, 47(3), 351 - 9
Optically active antifungal azoles . VIII . Synthesis and antifungal activity of 1-{(1R,2R)-2-(2,4-difluoro- and 2-fluorophenyl)-2-hydroxy-1-methyl-3-(1H-1,2,4-triazol-1-yl)propyl}- 3-(4-substituted phenyl)-2(1H,3H)-imidazolones and 2-imidazolidinones; Kitazaki T et al.; New optically active antifungal azoles, 1-{(1R,2R)-2-(2,4-difluoro- and 2-fluorophenyl)-2-hydroxy-1-methyl-3-(1H-1,2,4-triazol-1-yl)propyl }-3-(4- substituted phenyl)-2(1H,3H)-imidazolones (1,2) and 2-imidazolidinones (3,4), were prepared in a stereocontrolled manner from (1S)-1-{(2R)-2-(2,4- difluoro- and 2-fluorophenyl)-2-oxiranyl}ethanols (15, 16) . Compounds 1-4 showed potent antifungal activity against Candida albicans in vitro and in vivo, as well as a broad antifungal spectrum for various fungi in vitro . Furthermore, the imidazolidinones, 3b--e and 4d, e, were found to exert extremely strong growth-inhibitory activity against Aspergillus fumigatus.

Berl Munch Tierarztl Wochenschr, 1999 Mar, 112(3), 104 - 7
{Fungal flora in chicken stalls and its etiopathogenic importance for humans and animals}; Vissiennon T; In order to find out the mycoflora prevailing in chicken pens, and to appreciate the health hazards for employees (incl . veterinarians) and animals, twenty litter samples and 6 sedimented dust trials were analysed mycologically . The following results were found: Bedding and dust samples all contained between 3.57 and 1.30 x 10(7) c.f.u./g DW . The commonest fungus is A . fumigatus with 3.41 x 10(4) - 1.30 x 10(7) c.f.u./g DW in bedding and 2.70 x 10(5) - 3.30 x 10(6) c.f.u./g DW in sedimented dust . In addition, the following fungal spp . were detected (in c.f.u./g DW): Scopulariopsis brevicaulis (3.57 x 10(3) - 6.50 x 10(7)), Pseudallescheria boydii (2.70 x 10(6)), A . flavus group (up to 8.41 x 10(5)), A . clavatus (6.00 x 10(5)), Candida albicans (4.88 x 10(5)), Alternaria spp . (4.49 x 10(5)), Penicillium spp . (up to 1.06 x 10(5)), A . terreus (8.47 x 10(4)), A . versicolor group (up to 8.44 x 10(4)), A . niger (5.93 x 10(4)), Aureobasidium sp . (3.96 x 10(3)), Mucor circinelloides (3.42 x 10(3)), Phialophora sp . (1.30 x 10(3)), C . pseudoiropicalis (4.04 x 10(2)), A . nidulans (3.90 x 10(2)), Acremonium sp . (3.55 x 10(2)), M . racemosus (1.19 x 10(2)) . Negligible was the detected level of Chrysosporium pannorum (84.10), Rhizomucor pusillus (81.00), Beauveria alba (59.60), C . tropicalis (40.40), Trichoderma sp . (4.70), A . flavipes (4.38) . Dermatophytes were not found . The fungal spectrum was broader in the dust than in litter samples.

Biochim Biophys Acta, 1999 Apr 12, 1431(1), 107 - 19
Candidacidal activity prompted by N-terminus histatin-like domain of human salivary mucin (MUC7)1; Gururaja TL et al.; Histidine-rich peptides (histatins, Hsn) in saliva are thought to provide a non-immune defense against Candida albicans . Sequence homology search of the human salivary mucin, MUC7, against histatins revealed a domain at the N-terminus (R3-Q17) having 53% identity to Hsn-5 . To determine its candidacidal activity, this 15 residue basic histidine-rich domain of MUC7 (I) was prepared by solid-phase Fmoc chemistry . Various N- and C-terminal protected derivatives of I were also synthesized to correlate the effect of peptide overall charge in exhibiting cidal potency . Candidacidal activity measurement of I and its variants showed considerable ED50 values (effective dosage required to kill 50% of candida cells), albeit greater than Hsn-5 (ED50 approximately 4-6 microM) . Of the various analogs tested, N-terminal free acid (I, ED50 approximately 40 microM) and amide (V, ED50 approximately 16 microM) exhibited appreciable candidacidal activities suggesting the possible role of peptide net charge in cidal action . Blocking of N-terminus with a bulky octanoyl group showed only marginal effect on the cidal activity of I or V, indicating that hydrophobicity of these synthetic constructs may not be important for exerting such activities . Membrane-induced conformational transition from random coil to helical structures of all the test peptides implied their tendency to adapt order structures at the lipid-membrane interface similar to that of Hsn-5 . However, comparison of propensity for helical structure formation vs . ED50 indicated that cidal potency of MUC7 Hsn-like peptides depends largely on electrostatic interactions irrespective of secondary structural elements . Delineation of solution structure of the most active peptide (V) by 2D-NMR revealed essentially a non-structured conformation in aqueous medium, which further supported the fact that the peptide helical structure may not be a prerequisite for posing candidacidal activity . The formation of smaller truncated peptides and/or Hsn-like fragments on proteolytic degradation of intact MUC7 in the presence of oral flora provided indirect evidence that mucin could serve as a backup candidacidal agent to salivary Hsn.

J Comp Pathol, 1999 May, 120(4), 369 - 81
Inhibition of phagosome-lysosome fusion in ovine polymorphonuclear leucocytes by Ehrlichia (Cytoecetes) phagocytophila; Gokce HI et al.; Ehrlichia (Cytoecetes) phagocytophila, the causative agent of tick-borne fever, is an intracellular bacterium that survives and multiplies within granulocytes and monocytes . In the present study, the possible fusion of lysosomes with phagosomes containing E . phagocytophila was investigated in poly-morphonuclear (PMN) cells of sheep infected with the agent, acid phosphatase cytochemistry and cationized ferritin being used as markers of primary and secondary lysosomal enzymes . Latex beads or Candida albicans were incubated with infected and uninfected PMN cells and labelled with the same lysosomal markers . Lysosomal enzymes labelled with the markers were commonly found in phagosomes containing latex beads or C . albicans, but there was no evidence of phagosome-lysosome (P-L) fusion in phagosomes containing E . phagocytophila . It was significant that in cells that contained E . phagocytophila, latex beads and C . albicans, P-L fusion occurred only in phagosomes containing latex beads or C . albicans . However, evidence of P-L fusion with phagosomes containing E . phagocytophila was obtained when PMN cells were incubated with oxytetracycline, which is known to inhibit synthesis of bacterial proteins . These findings indicate that E . phagocytophila is capable of inhibiting P-L fusion and that oxytetracycline depresses this capability .

Yeast, 1999 Mar 15, 15(4), 323 - 7
Isolation and sequence of the GCR3 homologue from Candida albicans by complementation of (delta)gcr3 strain of Saccharomyces cerevisiae; Uemura H et al.; To study the function of GCR3, a gene involved in the expression of glycolytic genes in Saccharomyces cerevisiae, a Candida albicans gene which complements the growth defect of the (delta)gcr3 mutant was isolated . Transformants of this gene also recovered the glycolytic enzyme activities . Its DNA sequencing predicted an 886 amino acid protein with 30.4% identity to the Gcr3p of S . cerevisiae.

Eur J Pediatr, 1999 Apr, 158(4), 275 - 80
Fungal endocarditis in critically ill children; Aspesberro F et al.; All cases of infective endocarditis occurring from January 1990 to December 1996 at our institution were reviewed, with a special focus on fungal endocarditis . Five critically ill children with fungal endocarditis and eleven children with bacterial endocarditis were recorded . The proportion of fungal endocarditis in our series was 5/16 (31%) and Candida albicans (4/5) was the most common fungal pathogen . Only one patient required heart surgery because of a loose patch but all the others were treated only by medical management for cure . The hospital survival rate was 80% (4/5) and the overall long-term survival rate was 60% (3/5) with only one death directly related to fungal infection . CONCLUSION: Despite the small number of cases, a sole medical approach including amphotericin B and long-term fluconazole prophylaxis for the treatment of fungal endocarditis in critically ill children seems to offer an alternative to surgical treatment which may be kept for failure of medical treatment.

J Clin Microbiol, 1999 May, 37(5), 1464 - 8
Coaggregation of Candida dubliniensis with Fusobacterium nucleatum; Jabra-Rizk MA et al.; The binding of microorganisms to each other and oral surfaces contributes to the progression of microbial infections in the oral cavity . Candida dubliniensis, a newly characterized species, has been identified in human immunodeficiency virus-seropositive patients and other immunocompromised individuals . C . dubliniensis phenotypically resembles Candida albicans in many respects yet can be identified and differentiated as a unique Candida species by phenotypic and genetic profiles . The purpose of this study was to determine oral coaggregation (CoAg) partners of C . dubliniensis and to compare these findings with CoAg of C . albicans under the same environmental conditions . Fifteen isolates of C . dubliniensis and 40 isolates of C . albicans were tested for their ability to coaggregate with strains of Fusobacterium nucleatum, Peptostreptococcus micros, Peptostreptococcus magnus, Peptostreptococcus anaerobius, Porphyromonas gingivalis, and Prevotella intermedia . When C . dubliniensis and C . albicans strains were grown at 37 degrees C on Sabouraud dextrose agar, only C . dubliniensis strains coaggregated with F . nucleatum ATCC 49256 and no C . albicans strains showed CoAg . However, when the C . dubliniensis and C . albicans strains were grown at 25 or 45 degrees C, both C . dubliniensis and C . albicans strains demonstrated CoAg with F . nucleatum . Heating the C . albicans strains (grown at 37 degrees C) at 85 degrees C for 30 min or treating them with dithiothreitol allowed the C . albicans strains grown at 37 degrees C to coaggregate with F . nucleatum . CoAg at all growth temperatures was inhibited by mannose and alpha-methyl mannoside but not by EDTA or arginine . The CoAg reaction between F . nucleatum and the Candida species involved a heat-labile component on F . nucleatum and a mannan-containing heat-stable receptor on the Candida species . The CoAg reactions between F . nucleatum and the Candida species may be important in the colonization of the yeast in the oral cavity, and the CoAg of C . dubliniensis by F . nucleatum when grown at 37 degrees C provides a rapid, specific, and inexpensive means to differentiate C . dubliniensis from C . albicans isolates in the clinical laboratory.

Biochem Mol Biol Int, 1999 Mar, 47(3), 369 - 76
Structural characteristics of tenecin 3, an insect antifungal protein; Lee YT et al.; Tenecin 3, an antifungal protein, previously isolated from the insect Tenebrio molitor, inhibits growth of the fungus Candida albicans . However, the antifungal mechanism and functions of tenecin 3 remain unknown . As an initial step to study the mechanism and functions, physical and structural properties of tenecin 3 were examined by circular dichroism (CD) analysis and 2D nuclear overhauser effect spectroscopy . These analyses suggest that tenecin 3 has a propensity of random structure with very loose turn-like elements . The CD results also indicate that this random structural propensity is not significantly affected by temperature, pH, and by the presence of organic solvents or sodium dodecyl sulfate (SDS) micelles . However, the hydrodynamic studies suggest that tenecin 3 is not in extended form in spite of its random structural feature.

J Clin Microbiol, 1999 May, 37(5), 1587 - 90
Rapid PCR test for discriminating between Candida albicans and Candida dubliniensis isolates using primers derived from the pH-regulated PHR1 and PHR2 genes of C . albicans; Kurzai O et al.; The development of a satisfactory means to reliably distinguish between the two closely related species Candida albicans and Candida dubliniensis in the clinical mycology laboratory has proved difficult because these two species are phenotypically so similar . In this study, we have detected homologues of the pH-regulated C . albicans PHR1 and PHR2 genes in C . dubliniensis . Restriction fragment length polymorphism analysis suggests that there are significant sequence differences between the genes of the two species . In order to exploit this apparent difference, oligonucleotide primers based on the coding sequence of the C . albicans PHR1 structural gene were designed and used in PCR experiments . Use of these primers with C . albicans template DNA from 17 strains yielded a predicted 1.6-kb product, while C . dubliniensis template DNA from 19 strains yielded no product . We therefore propose that PCR using these primers is a rapid and reliable means of distinguishing the two germ tube- and chlamydospore-producing species C . albicans and C . dubliniensis.

J Clin Microbiol, 1999 May, 37(5), 1510 - 7
New enzyme immunoassays for sensitive detection of circulating Candida albicans mannan and antimannan antibodies: useful combined test for diagnosis of systemic candidiasis; Sendid B et al.; Two standardized enzyme immunoassays for the serological diagnosis of candidiasis were developed . The first one detects antimannan antibodies, while the second one detects mannan with a sensitivity of 0.1 ng/ml . These tests were applied to 162 serum samples retrospectively selected from 43 patients with mycologically and clinically proven candidiasis caused by Candida albicans . Forty-three serum samples were positive for mannan, and 63 had significant antibody levels . Strikingly, only five serum samples were simultaneously positive by both tests . When the results were analyzed per patient, 36 (84%) presented at least one serum positive by one test . For 30 of them, positivity by one test was always associated with negative results by the other test for any of the tested sera . For six patients whose sera were positive for either an antigen or an antibody response, a balance between positivity by each test was evidenced by kinetic analysis of sera drawn during the time course of the infection . Controls consisted of 98 serum samples from healthy individuals, 93 serum samples from patients hospitalized in intensive care units, and 39 serum samples from patients with deep mycoses . The sensitivities and specificities were 40 and 98% and 53 and 94% for mannanemia or antibody detection, respectively . These values reached 80 and 93%, respectively, when the results of both tests were combined . These observations, which clearly demonstrate a disparity between circulation of a given mannan catabolite and antimannan antibody response, suggest that use of both enzyme immunoassays may be useful for the routine diagnosis of candidiasis.

J Clin Microbiol, 1999 May, 37(5), 1398 - 403
Postsurgical Candida albicans infections associated with an extrinsically contaminated intravenous anesthetic agent; McNeil MM et al.; From 16 to 30 April 1990, four of 364 (1%) postsurgical patients at one hospital developed Candida albicans fungemia or endophthalmitis . The case patients' surgeries were clustered on two days . To identify risk factors for C . albicans infections, we conducted a cohort study comparing these 4 patients with 67 control patients who had surgeries on the same days but did not acquire C . albicans infections . The participation of anesthesiologist 9 (relative risk {RR}, undefined; P < 0.001) and receipt of intravenous propofol, an anesthetic agent without preservative, which was administered by an infusion pump (RR, 8.8; P = 0.048) were identified as risk factors for C . albicans infections . The anesthetic had been recently introduced in the hospital . Hand cultures of 8 of 14 (57%) anesthesiologists were positive for Candida species; one yielded C . albicans . Anesthesiologist 9 was the only one to use stored syringes of propofol in the infusion pump and to reuse propofol syringes . DNA fingerprinting with a digoxigenin-labeled C . albicans repetitive element 2 probe and electrophoretic karyotyping showed two distinct banding patterns among patient isolates . We hypothesize that extrinsic contamination of propofol by anesthesiologist 9 likely resulted in C . albicans infections . These data suggest that strict aseptic techniques must be used when preparing and administering propofol.

J Clin Microbiol, 1999 May, 37(5), 1376 - 80
High aspartyl proteinase production and vaginitis in human immunodeficiency virus-infected women; de Bernardis F et al.; Vaginal isolates of Candida albicans from human immunodeficiency virus-positive (HIV+) and HIV- women with or without candidal vaginitis were examined for secretory aspartyl proteinase (Sap) production in vitro and in vivo and for the possible correlation of Sap production with pathology and antimycotic susceptibility in vitro . HIV+ women with candidal vaginitis were infected by strains of C . albicans showing significantly higher levels of Sap, a virulence enzyme, than strains isolated from HIV+, C . albicans carrier subjects and HIV- subjects with vaginitis . The greater production of Sap in vitro was paralleled by greater amounts of Sap in the vaginal fluids of infected subjects . In an estrogen-dependent, rat vaginitis model, a strain of C . albicans producing a high level of Sap that was isolated from an HIV+ woman with vaginitis was more pathogenic than a strain of C . albicans that was isolated primarily from an HIV-, Candida carrier . In the same model, pepstatin A, a strong Sap inhibitor, exerted a strong curative effect on experimental vaginitis . No correlation was found between Sap production and antimycotic susceptibility, as most of the isolates were fully susceptible to fluconazole, itraconazole, and other antimycotics, regardless of their source (subjects infected with strains producing high or low levels of Sap, subjects with vaginitis or carrier subjects, or subjects with or without HIV) . Thus, high Sap production is associated with virulence of C . albicans but not with fungal resistance to fluconazole in HIV-infected subjects, and Sap is a potentially new therapeutic target in candidal vaginitis.

J Immunol, 1999 Apr 15, 162(8), 4876 - 81
Soluble murine IL-1 receptor type I induces release of constitutive IL-1 alpha; Netea MG et al.; IL-1 alpha and IL-1 beta are proinflammatory cytokines involved in the pathogenesis of many infectious and noninfectious inflammatory diseases . To reduce IL-1 toxicity, extracellular domains of the soluble (s) IL-1R are shed from cell membranes and prevent triggering of cell-bound receptors . We investigated to what extent murine sIL-1RI can neutralize the IL-1 produced by LPS-stimulated macrophages . When mouse peritoneal macrophages were incubated with LPS, addition of sIL-1RI significantly inhibited the bioactivity of IL-1 . Stimulation of cells with sIL-1RI alone induced no bioactive IL-1 . When immunoreactive cytokine concentrations were measured with specific radioimmunoassays, sIL-1RI alone appeared to induce a significant release of IL-1 alpha in a concentration-dependent manner . This effect was independent of new protein synthesis . The production of IL-1 beta or TNF-alpha was not influenced by sIL-1RI . There was no interference of sIL-1RI with the IL-1 alpha radioimmunoassay . In mice, an i.v . injection of sIL-RI alone induced a rapid release of IL-1 alpha, but not of TNF-alpha or IL-1 beta . Treatment of mice with sIL-1RI improved the survival during a lethal infection with Candida albicans . In conclusion, sIL-1RI induces a rapid release of IL-1 alpha from cells, as well as into the systemic circulation . Although this IL-1 alpha may be inactivated in circulation by the same sIL-1RI, this phenomenon probably has immunostimulatory effects at local levels where the sIL-1RI-induced IL-1 alpha acts in a paracrine or autocrine manner.

Med Mycol, 1999 Feb, 37(1), 35 - 41
In vitro susceptibility of Candida species to lactoferrin; Xu YY et al.; Lactoferrin is an antimicrobial protein present in human mucosal secretions as well as saliva . As there is no information on the relative fungicidal activity of human and bovine lactoferrin, an oral isolate of Candida albicans was studied for its susceptibility to these two proteins . Exposure to a concentration of 20 micrograms ml-1 of either HLF or BLF at 37 degrees C inactivated the yeast to the same degree irrespective of the incubation time of 45, 90 or 150 min . A similar study, using 20 micrograms ml-1 BLF and an incubation time of 150 min, elicited varying anticandidal activity against 35 isolates belonging to six different Candida species . Thus, BLF was fungicidal for the six Candida species in the following decreasing order, C . tropicalis > C . krusei > C . albicans > C . guilliermondii > C . parapsilosis > C . glabrata; the latter being the most resistant . These Candida species also demonstrated significant intra-species variation in susceptibility to the protein (P < 0.05) . When the yeast cells exposed to BLF were examined by cryo-scanning electron microscopy, profound cell wall changes such as cell surface blebs, swelling and cell collapse were noted . These findings suggest that lactoferrin, a constituent of saliva, may differentially modulate the carriage of Candida species in the oral cavity.

Quintessence Int, 1998 Nov, 29(11), 705 - 10
Candida albicans levels in patients with Sjögren's syndrome before and after long-term use of pilocarpine hydrochloride: a pilot study; Rhodus NL et al.; OBJECTIVE: The purpose of this study was to compare the quantities of oral Candida albicans in patients with primary and secondary Sjogren's syndrome before and after the use of orally administered pilocarpine hydrochloride for 1 year . METHOD AND MATERIALS: Twelve female subjects with primary (n = 4) and secondary (n = 8) Sjogren's syndrome (mean age +/- SEM = 56.7 +/- 5.7 years) were enrolled in the study, after meeting rigid enrollment criteria . Oropharyngeal collection of samples and culturing was performed on each subject . Cultures specific for Candida albicans were plated into a culture media tube using the Oricult kit and also by serial dilutions and plating by a streptomycin-vancomycin technique . Cultures were incubated for 48 hours at 37 degrees C . The subjects used 5 mg of pilocarpine hydrochloride, administered orally three times daily, for 1 year, after which both of the Candida cultures were repeated . None of the subjects used antifungal medications, none smoked, and all were dentate . RESULTS: There was a significant difference in the prevalence of Candida after the use of pilocarpine hydrochloride for both groups . At the start of the study, 75% of all subjects were positive for Candida . Following the use of pilocarpine, 25% had positive cultures . There was also a decrease in the prevalence of clinical manifestations of infection from 75% of subjects to 25% . There was a significant decrease in the numbers of Candida cultured following the use of pilocarpine . CONCLUSION: Long-term administration of pilocarpine hydrochloride resulted in a significant reduction in Candida albicans colonization in patients with primary or secondary Sjogren's syndrome.

Yonsei Med J, 1999 Feb, 40(1), 56 - 60
Diagnosis of trichomoniasis by polymerase chain reaction; Ryu JS et al.; The clinical usefulness of polymerase chain reaction (PCR) for the diagnosis of trichomoniasis was evaluated in comparison with other conventional tests . PCR was used for specific detection of Trichomonas vaginalis by primers based on the repetitive sequence cloned from T . vaginalis (TV-E650) . Between June 1996 and August 1997, 426 patients visited the department of obstetrics and gynecology, Hanyang University Kuri Hospital and were examined for trichomoniasis using wet mount examination, Papanicolaou (Pap) smear, culture and PCR . One hundred and seventy-seven patients (group A) visited with the symptoms of vaginal discharge and 249 patients (group B) visited for regular cervical Pap smear with no vaginal symptoms . From group A (n = 177), 3 infections (2.0%) were detected by wet mount, 6 infections (3.3%) by Pap smear and culture, and 17 infections (10.4%) by PCR . From group B (n = 249), 4 patients (1.6%) were found to have T . vaginalis by culture and 6 infections (2.4%) were detected by PCR . Therefore, in both groups, PCR for T . vaginalis showed a higher detection rate compared with conventional wet mount, Pap smear or culture . The detection by PCR was specific for T . vaginalis since no amplification was detected with DNAs from other protozoa and Candida albicans . The sensitivity and specificity of PCR were 100% . This method could detect T . vaginalis in vaginal discharge at a concentration as low as 1 cell per PCR mixture . These results indicate that PCR could be used as a specific and sensitive diagnostic tool for human trichomoniasis.

Int J Prosthodont, 1999 Jan-Feb, 12(1), 83 - 6
Adherence of Candida albicans to the surface of polymethylmethacrylate--E glass fiber composite used in dentures; Waltimo T et al.; PURPOSE: The use of reinforcing fibers in dentures has raised concerns about possible increased adherence of yeasts to the surface . The aim of this in vitro study was to compare the adherence of Candida albicans to the surface of denture-base polymer and to E-glass fibers . MATERIALS AND METHODS: Test specimens were made from an autopolymerized denture-base resin (Palapress) reinforced with preimpregnated unidirectional E-glass fibers, which were exposed at the surface . The test specimens were pretreated with parotid saliva and incubated without agitation in standardized yeast suspensions (10(8) colony-forming units per mL) in phosphate-buffered saline at 37 degrees C for 1 hour . The test specimens were then washed to remove nonadherent cells . After being air dried, they were sputter coated with gold-palladium for scanning electron microscopy (SEM) . To compare the adherence to different surfaces, the number of yeast cells found either on the polymer matrix or on the glass fibers was counted from SEM fields (170 microns x 120 microns, 600 x) of randomly selected areas . RESULTS: The mean density of yeast cell found on the surface of the polymer matrix was significantly higher (P < 0.001) than that on the surface of glass fibers . The number of adherent yeast cells found at the interface between the fibers and polymer matrix was high . CONCLUSION: The adherence of C albicans to E-glass fibers was lower than to polymer matrix in the denture composite . If fibers are exposed only during polishing of the composite, the reinforcing material appears not to increase the adherence of this common oral yeast . However, areas with permanently exposed fibers may provide mechanical retention for yeast cells at the interface of the components.

J Appl Microbiol, 1999 Mar, 86(3), 446 - 52
Influence of organic matter, cations and surfactants on the antimicrobial activity of Melaleuca alternifolia (tea tree) oil in vitro; Hammer KA et al.; The effect of some potentially interfering substances and conditions on the antimicrobial activity of Melaleuca alternifolia (tea tree) oil was investigated . Agar and broth dilution methods were used to determine minimum inhibitory and cidal concentrations of tea tree oil in the presence and absence of each potentially interfering substance . Activity was determined against Gram-positive and -negative bacteria, and Candida albicans . Minimum inhibitory or cidal concentrations differed from controls by two or more dilutions, for one or more organisms, where Tween-20, Tween-80, skim-milk powder and bovine serum albumin were assessed . These differences were not seen when assays were performed in anaerobic conditions, or in the presence of calcium and magnesium ions . The effect of organic matter on the antimicrobial activity of tea tree oil was also investigated by an organic soil neutralization test . Organisms were exposed to lethal concentrations of tea tree oil ranging from 1-10% (v/v), in the presence of 1-30% (w/v) dry bakers' yeast . After 10 min contact time, viability was determined . At > or = 1%, organic matter compromised the activity of each concentration of tea tree oil against Staphylococcus aureus and C . albicans . At 10% or more, organic matter compromised the activity of each tea tree oil concentration against Pseudomonas aeruginosa . Organic matter affected 1 and 2% tea tree oil, but not 4 and 8%, against Escherichia coli . In conclusion, organic matter and surfactants compromise the antimicrobial activity of tea tree oil, although these effects vary between organisms.

Clin Exp Immunol, 1999 Mar, 115(3), 491 - 7
Candida albicans suppresses nitric oxide (NO) production by interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS)-stimulated murine peritoneal macrophages; Chinen T et al.; We examined the in vitro effect of Candida albicans on NO production by macrophages . Candida albicans suppressed not only NO production but also expression of inducible NO synthase (iNOS) mRNA by murine IFN-gamma and bacterial LPS-stimulated peritoneal macrophages . The suppression was not associated with inhibition but rather stimulation of IL-1 beta production . This effect was observed when more than 1 x 10(3)/ml of Candida albicans were added to macrophage cultures (1 x 10(6) cells/ml) and reached a maximal level at 1 x 10(6)/ml . The NO inhibitory effect of Candida albicans was mediated predominantly by as yet unidentified soluble factor(s) and to a lesser extent by direct contact . In addition, heat- or paraformaldehyde-killed Candida albicans did not show this inhibitory activity . Culture supernatant of Candida albicans also inhibited NO production by activated macrophages in a dose-dependent manner, and increased IL-1 beta production . Finally, the inhibitory effect was not mediated by IL-10 and transforming growth factor-beta (TGF-beta), since neutralizing antibodies to these cytokines did not influence Candida albicans-induced reduction in macrophage NO production . Our results suggest that Candida albicans may evade host defence mechanism(s) through a soluble factor-mediated suppression of NO production by stimulated macrophages, and that the effect is independent of production of immunosuppressive cytokines such as IL-10 and TGF-beta.

Thorax, 1998 Aug, 53 Suppl 2, S47 - 51
Role of the indoor environment in determining the severity of asthma; Woodcock A et al.; Allergen exposure can confound the management of asthma . To understand the potential mechanisms by which allergens increase the steroid requirements in atopic asthmatics, we examined the effects of allergens on glucocorticoid receptor (GCR) binding affinity and glucocorticoid (GC) responsiveness of peripheral blood mononuclear cells (PBMC) from atopic asthmatics . A significant reduction (p < 0.001) in the GCR binding affinity (Kd) was observed in ragweed-allergic asthmatics during ragweed pollen season compared with PBMC obtained before and after ragweed season . In vitro effects of allergen on PBMC GCR Kd were also examined by incubating PBMC from atopic asthmatics with allergen (ragweed and cat) versus Candida albicans . GCR binding affinity was significantly reduced after incubation with ragweed (p < 0.001) or cat allergen (p < 0.001) compared with baseline or C . albicans stimulation . This effect was limited to atopic asthmatics in that in vitro cat allergen incubation for 48 h failed to significantly alter GCR binding affinity in nonasthmatic, atopic individuals . These allergen-induced reductions in GCR binding affinity also rendered the PBMC less sensitive to the inhibitory effects of hydrocortisone and dexamethasone on allergen-induced proliferation (p < 0.01) . To test the hypothesis that allergen-induced alterations in GCR binding affinity were cytokine-induced, we examined the effects of interleukin-2 (IL-2) and IL-4 neutralization using anticytokine antibodies . Addition of both anti-IL-2 and anti-IL-4 antibodies resulted in a significant (p < 0.001) inhibition of allergen-induced alterations in GCR binding affinity . Furthermore incubation with cat allergen induced significantly higher concentrations of IL-2 (p = 0.03) and IL-4 (p = 0.02) by PBMC from atopic as compared with nonatopic subjects . Our current observations suggest that allergen exposure may contribute to poor asthma control by reducing GCR binding affinity in mononuclear cells . This appears to be mediated through IL-2 and IL-4 . These findings may have important implications for novel approaches to the treatment of poorly controlled asthma.

J Clin Pathol, 1998 Nov, 51(11), 857 - 9
Immunocytochemical detection of Candida albicans in formalin fixed, paraffin embedded material; Williams DW et al.; AIM: To assess the ability of the commercially available monoclonal antibody 1B12 (BioGenex, San Ramon, USA) to identify C albicans in formalin fixed, paraffin wax embedded material (FFPE) . METHODS: Broth cultures of 20 strains of seven Candida species were resuspended in 4% agarose blocks, fixed in formalin for 24 hours, and embedded in paraffin wax . In addition, 16 blocks of FFPE tissue known to contain periodic acid-Schiff positive fungal hyphae were examined . Antigen retrieval involved microwave treatment of specimens in citrate buffer (0.01 M; pH 6.5) before addition of 1B12 antibody for 24 hours . Bound antibody was subsequently detected using a biotinylated link antibody and a peroxidase conjugated streptavidin . RESULTS: Only C albicans strains were 1B12 positive in the agarose blocks . All FFPE tissue blocks were found to contain 1B12 positive hyphal structures, indicating the presence of C albicans . CONCLUSIONS: The ability to identify candida organisms penetrating the lesional tissue in cases of chronic hyperplastic candidosis will help to clarify the role of individual Candida spp in this important form of oral candidosis.

J Infect Dis, 1999 May, 179(5), 1301 - 4
Modulation of neutrophil-mediated activity against the pseudohyphal form of Candida albicans by granulocyte colony-stimulating factor (G-CSF) administered in vivo; Gaviria JM et al.; Renewed interest in neutrophil transfusions has emerged with the development and clinical use of granulocyte colony-stimulating factor (G-CSF) . G-CSF not only increases neutrophil (polymorphonuclear leukocyte, PMNL) production but also modulates various physiological properties of PMNL . The effects of G-CSF on PMNL-mediated fungicidal activity were evaluated by administration of G-CSF (300 micrograms/day subcutaneously) to 5 healthy volunteers for 6 days . G-CSF significantly enhanced PMNL-mediated damage of Candida albicans pseudohyphae by 33% (P=.007) on day 2 and by 44% (P=.04) on day 6 at a 10:1 effector:target ratio . In contrast, the ability of PMNL to induce damage of hyphae from either Fusarium solani or Aspergillus fumigatus did not significantly change during the study period . These data demonstrate that G-CSF administered in vivo modulates PMNL-mediated fungicidal activity against the pseudohyphal form of C . albicans, thereby suggesting potential utility of G-CSF as a biologic response-modifying therapy in some opportunistic fungal infections.

Eur J Clin Microbiol Infect Dis, 1999 Jan, 18(1), 59 - 61
Susceptibility of Candida species isolated from female prostitutes with vulvovaginitis to antifungal agents and boric acid; Otero L et al.; The aim of this study was to determine the antifungal susceptibility of 108 Candida albicans and 23 Candida glabrata isolates obtained from female prostitutes with vulvovaginitis, a population for which available data is limited . Amphotericin B, flucytosine, and fluconazole were tested by broth microdilution, and boric acid was tested by the agar dilution method . The susceptibility patterns found in this population were the same as those in the general population . Candida glabrata required greater concentrations of boric acid for inhibition in vitro than did Candida albicans.

Int J Dermatol, 1999 Feb, 38(2), 119 - 21
Cutaneous infections by papillomavirus, herpes zoster and Candida albicans as the only manifestation of idiopathic CD4+ T lymphocytopenia; Manchado Lopez P et al.; BACKGROUND: Selective depletion of CD4+ T lymphocytes is common in both primary and secondary immunodeficiencies . Idiopathic CD4+ T lymphocytopenia (ICL) cases are defined as a persistent CD4+ T lymphocyte count of less than 300x10(6) cells/L and/or less than 20% of the total T-cell count . METHOD: A 40-year-old woman, with a history of psoriasis and paracetamol allergy, presented with persistent warts of the hands and condylomas of the ano-genitalia . Histological and virological analysis was carried out on genital and cutaneous lesions and peripheral blood . RESULTS: Serology for HIV-1, HIV-2, Epstein-Barr virus and parvovirus B19 were negative . There was lymphopenia of 10% CD4+ cells, with normal numbers of total leukocytes; there were no other-abnormal immunological findings . DNA analysis of cutaneous lesions revealed HPV-49 and HPV-3 in the hands and HPV-6 in the genital region . CONCLUSIONS: The cause of the ICL in this patient is unknown . HPV is not known to be an immunosuppressive agent; it remains to be determined whether the HPV-associated lesions are the cause or the result of immunosuppression.

Med Clin (Barc), 1999 Feb 20, 112(6), 211 - 4
{Epidemiology of yeast colonization and oropharyngeal infection other than Candida albicans in patients with HIV infection}; Masia Canuto MM et al.; BACKGROUND: An increasing frequency of opportunistic fungal infections in immunosuppressed patients in recent years . Concurrent with this finding, it has been noted an increasing use of fluconazole . In addition, non-Candida albicans species (NCAS), most of which are fluconazole-resistant have been increasing isolated . The aim of this study was to investigate the epidemiology of colonization and infection due to NCAS in HIV-infected patients . PATIENTS AND METHODS: A cross sectional study was conducted with HIV-infected patients in different stages, who were attended at two hospitals in Alicante, Spain . We assessed the prevalence and microbiology of oropharyngeal colonization and infection due to Candida spp., and its fluconazole susceptibility patterns . To determine the clinical risk factors for the development of fluconazole resistance, we carried out a case-control study with prevalent cases . RESULTS: We studied 168 strains from 153 patients . NCAS were isolated in 32 (21%) of them, 25 (77%) were colonized, and 5 (26%) had infection due to NCAS . The most common isolate was Candida glabrata (n = 15) . MICs were significantly higher for NCAS than for Candida albicans species, with a MIC50 of 16 and 0.25 microgram/ml, respectively, and a MIC90 of 128 micrograms/ml and 8 micrograms/ml (p = 0.0001) . The median CD4 cell count in patients with NCAS was 0.06 x 10(9)/l, and 0.19 x 10(9)/l patients with Candida albicans (p = 0.009) . Overall, 56% of the patients with NCAS and 41% of the patients with Candida albicans had been treated with fluconazole (p = 0.1) . CONCLUSIONS: NCAS are isolated in a high proportion of HIV infected patients . Most of the NCAS have a decreased susceptibility to fluconazole . The only risk factor associated with the acquisition of NCAS in HIV-infected patients is an advanced immunosuppression.

Phytother Res, 1999 Mar, 13(2), 151 - 6
Antimicrobial triterpenes from Ilex integra and the mechanism of antifungal action; Haraguchi H et al.; Antimicrobial triterpenes were isolated from the fruits of Ilex integra . Their structures were elucidated by spectral data and identified as rotundic acid (1), ulsolic acid (2) and peduncloside (3) . Triterpene 1 showed significantly broad antimicrobial activity against bacteria, yeast and filamentous fungi . The antifungal activity of 1 was reversed by fatty acids . Cellular constituents leaked from Candida albicans cells incubated with triterpene 1 . These results suggest that the antimicrobial activity of triterpenes in I . integra is due to a change of membrane permeability arising from membrane lipid alteration.

Hosp Pharm, 1993 Jan, 28(1), 11 - 3, 16-8
Evaluation of an aseptic technique testing and challenge kit (ATTACK); Turco S et al.; Currently, there is considerable discussion and concern about quality assurance when sterile pharmaceuticals are prepared by pharmacists and other health professionals . This study describes the utility of a kit made by Marsam Pharmaceutics Inc . in detecting microbial contamination during simulated drug transfers by syringe . Common microbial detection techniques were incorporated in a simple series of transfers intended to simulate actual use . Growth promotion studies were carried out using Trypticase Soy Broth initially in tubes and then in vials (as supplied in the kit) . Three test organisms were employed (Staphylococcus epidermidis, Bacillus subtilis, and Candida albicans) and inoculated at three levels, < 1000, < 100, and < 10 CFU/mL . All studies demonstrated growth occurring in 100% of the trials in 8 days . Similar studies were initiated in stored media (32 month) to determine the effects of time on the ability of the medium to support growth . Growth promoting ability of 32 month old media showed no significant difference . 100% of the vials inoculated showed growth in 8 days . Once the growth promoting qualities of the medium and vials was established a kit was developed then a study to determine the effectiveness of the kit as used was undertaken . Kits were manipulated according to the directions for use by a trained individual, under aseptic conditions and by an untrained individual in an area described as less than desirable . No growth occurred in the vials of the 10 kits used to illustrate good technique and good environmental conditions with 30% (3 of 10) of the kits showing contamination when transfers by syringe were carried out in the less than desirable setting.(ABSTRACT TRUNCATED AT 250 WORDS)

Antimicrob Agents Chemother, 1999 Apr, 43(4), 830 - 5
LY303366 exhibits rapid and potent fungicidal activity in flow cytometric assays of yeast viability; Green LJ et al.; LY303366 is a semisynthetic analog of the antifungal lipopeptide echinocandin B that inhibits (1,3)-beta-D-glucan synthase and exhibits efficacy in animal models of human fungal infections . In this study, we utilized flow cytometric analysis of propidium iodide uptake, single-cell sorting, and standard microbiological plating methods to study the antifungal effect of LY303366 on Saccharomyces cerevisiae and Candida albicans . Our data indicate that an initial 5-min pulse treatment with LY303366 caused yeasts to take up propidium iodide and lose their ability to grow . Amphotericin B and cilofungin required longer exposure periods (30 and 180 min, respectively) and higher concentrations to elicit these fungicidal effects . These two measurements of fungicidal activity by LY303366 were highly correlated (r > 0.99) in concentration response and time course experiments . As further validation, LY303366-treated yeasts that stained with propidium iodide were unable to grow in single-cell-sorted cultures . Our data indicate that LY303366 is potent and rapidly fungicidal for actively growing yeasts . The potency and rapid action of this new fungicidal compound suggest that LY303366 may be useful for antifungal therapy.

Antimicrob Agents Chemother, 1999 Apr, 43(4), 763 - 8
Effects of azole antifungal drugs on the transition from yeast cells to hyphae in susceptible and resistant isolates of the pathogenic yeast Candida albicans; Ha KC et al.; Oral infections caused by the yeast Candida albicans are some of the most frequent and earliest opportunistic infections in human immunodeficiency virus-infected patients . The widespread use of azole antifungal drugs has led to the development of drug resistance, creating a major problem in the treatment of yeast infections in AIDS patients and other immunocompromised individuals . Several molecular mechanisms that contribute to drug resistance have been identified . In C . albicans, the ability to morphologically switch from yeast cells (blastospores) to filamentous forms (hyphae) is an important virulence factor which contributes to the dissemination of Candida in host tissues and which promotes infection and invasion . A positive correlation between the level of antifungal drug resistance and the ability to form hyphae in the presence of azole drugs has been identified . Under hypha-inducing conditions in the presence of an azole drug, resistant clinical isolates form hyphae, while susceptible yeast isolates do not . This correlation is observed in a random sample from a population of susceptible and resistant isolates and is independent of the mechanisms of resistance . 35S-methionine incorporation suggests that growth inhibition is not sufficient to explain the inhibition of hyphal formation, but it may contribute to this inhibition.

Gene, 1999 Mar 18, 229(1-2), 183 - 91
Translation elongation factor 2 is encoded by a single essential gene in Candida albicans; Mendoza A et al.; Translation elongation factor 2 (eEF2) is a large protein of more than 800 amino acids which establishes complex interactions with the ribosome in order to catalyze the conformational changes needed for translation elongation . Unlike other yeasts, the pathogenic fungus Candida albicans was found to have a single gene encoding this factor per haploid genome, located on chromosome 2 . Expression of this locus is essential for vegetative growth, as evidenced by placing it under the control of a repressible promoter . This C . albicans gene, named EFT2, was cloned and sequenced (EMBL accession number Y09664) . Genomic and cDNA sequence analysis identified common transcription initiation and termination signals and an 842 amino acid open reading frame (ORF), which is interrupted by a single intron . Despite some genetic differences, CaEFT2 was capable of complementing a Saccharomyces cerevisiae Deltaeft1 Deltaeft2 null mutant, which lacks endogenous eEF2, indicating that CaEFT2 can be expressed from its own promoter and its intron can be correctly spliced in S . cerevisiae.

Eur J Biochem, 1999 Feb, 260(1), 217 - 26
Genetic and biochemical characterization of phosphofructokinase from the opportunistic pathogenic yeast Candida albicans; Lorberg A et al.; We have used the two PFK genes of Saccharomyces cerevisiae encoding the alpha and beta-subunit of the enzyme phosphofructokinase (Pfk) as heterologous probes to isolate fragments of the respective genes from the dimorphic pathogenic fungus Candida albicans . The complete coding sequences were obtained by combining sequences of chromosomal fragments and fragments obtained by inverse polymerase chain reaction (PCR) . The CaPFK1 and CaPFK2 comprise open reading frames of 2961 bp and 2838 bp, respectively, encoding Pfk subunits with deduced molecular masses of 109 kDa and 104 kDa . The genes presumably evolved by a duplication event from a prokaryotic type ancestor, followed by another duplication . Heterologous expression in S . cerevisiae revealed that each gene alone was able to complement the glucose-negative phenotype of a pfk1 pfk2 double mutant . In vitro Pfk activity in S . cerevisiae was not only obtained after coexpression of both genes, but also in conjunction with the respective complementary subunits from S . cerevisiae . This indicates the formation of functional hetero-oligomers consisting of C . albicans and S . cerevisiae Pfk subunits . In C . albicans, specific Pfk activity was shown to decrease twofold upon induction of hyphal growth . CaPfk cross-reacts with a polyclonal antiserum raised against ScPfk and displays similar allosteric properties, i.e . inhibition by ATP and activation by AMP and fructose 2,6-bisphosphate.

J Infect, 1999 Jan, 38(1), 51 - 3
Successful treatment of candidal osteomyelitis with fluconazole following failure with liposomal amphotericin B; Turner DL et al.; A case of multiple relapses of Candida albicans infection of deep tissues is described . Treatment was complicated by renal impairment, but therapy with a liposomal amphotericin product failed to eradicate the third recurrence which subsequently resolved after protracted exposure to oral fluconazole.

Support Care Cancer, 1999 Mar, 7(2), 100 - 2
Microbiological safety of essential oils used in complementary therapies and the activity of these compounds against bacterial and fungal pathogens; Maudsley F et al.; To determine the safety of plant essential oils we determined the sterility of eight of these products obtained from retail outlets . In addition, the ability of oils to support the growth of fungal and bacterial pathogens was examined . The antimicrobial activity of these products against seven bacterial species and Candida albicans was also investigated . All oils and their respective carriers were sterile . Methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa were unable to survive in oils for longer than 6 h, whereas C . albicans was able to survive, but not multiply, in ylang ylang oil for at least 48 h.

Mycoses, 1998, 41 Suppl 2, 31 - 6
{Vertical transmission of Candida and its consequences}; Blaschke-Hellmessen R; The subpartal transmission of Candida albicans from the vagina of the mother to the newborn is an old and often discussed problem . Thereby the decrease of the infection rate and the prevention of systemic mycoses due to Candida--especially in newborns of risk--are the main objectives . At the end of pregnancy C . albicans is found in vaginal secretions in 25-30% of the women . 70-85% of these women subpartally contaminate their infants with this yeast . Thus 22-24% of all infants acquire C . albicans sub partus . From this situation the following conclusions may be drawn: 1 . A prepartal prophylaxis for mycoses in pregnant women with vaginal Candida colonization is to obtain by an intravaginal treatment with polyene or azole antimycotics at the end of pregnancy . Recommendations are offered . 2 . A prophylaxis for mycoses in newborns which are especially disposed for systemic candidosis by several factors of risk is to initiate . The oral application of polyene antimycoticas during the considerable endangering by mycoses has been proved to be useful . Referring to this recommendations are offered.

Mycoses, 1998, 41 Suppl 2, 23 - 5
{Comparison of strain specificity of yeasts from various organs of women with vaginal candidiasis}; Mendling W et al.; Sexual partners harbour often identical yeast strains in the vagina, orointestinal tract and sperm in cases of recurrent vulvovaginal candidoses . Mycologic cultures from the vagina, mouth and stool of the patient and from the mouth and sperm of her partner were cultured on Sabouraud-Glucose-Agar and, if positive, specified by Candida ID-Agar (Biomerieux), rice agar and the system Walk away (Dade) . Equal species were compared by DNR-fingerprinting using PCR . The vagina of 22 women was in 21 cases infected by Candida albicans and in one case by Candida glabrata . The culture of mouth or stool of 18 women was in 11 cases identical with those of the vagina . In 13 cases of 18 sexual male partners Candida albicans was found being identical with the strain of the female partner in 8 cases . 4 of the identical strains were grown from the sperm . Future prospective investigations shall prove whether a consequent treatment of both partners to eradicate all identical yeasts is able to improve the treatment results in such women.

Infect Immun, 1999 Apr, 67(4), 1828 - 36
Severe impairment in early host defense against Candida albicans in mice deficient in myeloperoxidase; Aratani Y et al.; Myeloperoxidase (MPO) catalyzes the reaction of hydrogen peroxide with chloride ion to produce hypochlorous acid (HOCl), which is used for microbial killing by phagocytic cells . Despite the important role of MPO in host defense, however, MPO deficiency is relatively common in humans, and most of these individuals are in good health . To define the in vivo role of MPO, we have generated by gene targeting mice having no MPO activity in their neutrophils and monocytes . The mice without MPO developed normally, were fertile, and showed normal clearance of intraperitoneal Staphylococcus aureus . However, they showed increased susceptibility to pneumonia and death following intratracheal infection with Candida albicans . Furthermore, the lack of MPO significantly enhanced the dissemination of intraperitoneally injected C . albicans into various organs during the first 7 days . Thus, MPO is important for early host defense against fungal infection, and the inability to generate HOCl cannot be compensated for by other oxygen-dependent systems in vivo in mice . The mutant mice serve as a model for studying pulmonary and systemic candidiasis.

J Invest Dermatol, 1999 Mar, 112(3), 383 - 6
In vivo expression and localization of Candida albicans secreted aspartyl proteinases during oral candidiasis in HIV-infected patients; Schaller M et al.; Isoforms of aspartyl proteinase (Sap), which are encoded by at least nine related SAP genes, have been implicated to be a major virulence factor of the opportunistic yeast Candida albicans in experimental infections . Although it is generally assumed that proteinases are important for infections, detailed information on the pathogenetic role of Saps is still lacking . The same applies to the question whether the genes and corresponding isoforms of the enzyme are expressed during oral infection . For in vivo investigations, parts of the lesional oral epithelium were collected from three HIV-infected patients with oropharyngeal candidiasis . Immunoelectron microscopy was performed (pre- and post-embedding gold labeling with silver enhancement) using an anti-Sap murine monoclonal antibody directed against the gene products Sap1-3 . It was possible to demonstrate expression of Sap antigens in each of the three samples of human oral candidiasis . This suggests that at least one of the genes SAP1-3 was expressed at the time of sample collection . Furthermore, a possible role of the enzymes during the interaction of yeast cells and mucosal cells is suggested: the majority of Sap antigens is secreted by those C . albicans cells that adhere directly to the epithelial surface . Sap immunoreactivity can be detected in particular at the site of close contact between C . albicans and epithelial cells, suggesting a pathogenetic role of the Saps in host-fungal interaction . Thus, inhibition of the enzyme might prove to be an important alternative in the prevention and treatment of candidiasis.

J Oral Rehabil, 1999 Feb, 26(2), 177 - 82
Examining the secretor status in the saliva of patients with oral pre-cancerous lesions; Vidas I et al.; It has been demonstrated in a number of earlier studies on the aetiology and pathogenesis of certain diseases that the patients' secretor status (ABO (H) blood group antigens) may possibly be a factor influencing the development of systemic oral diseases . This likelihood has prompted the present study, to examine the differences in the saliva secretor status by comparing patients with oral pre-cancerous lesions on the one hand, and the healthy population on the other; (i) in relation to the intensity of the clinical manifestation of diseases and (ii) in relation to the intensity of epithelial dysplasia of patients with oral pre-cancerous lesions . In total 122 subjects were examined, half of whom suffered from oral pre-cancerous lesions (excluding Candida albicans in oral smears), while the other half were the healthy control group . All were subjected to clinical oral examinations and standard evaluation tests in order to establish the secretor status of their saliva . In the group of patients with oral pre-cancerous lesions (experimental group), a pathohistological examination of the oral mucosa was performed . The results have demonstrated that the large majority of the people examined in both groups were secretors and no significant difference between secretors and non-secretors was found in the comparison between the experimental group and the healthy control group . However, (i) we found a higher intensity of oral disease in the non-secretor group, and (ii) the occurrence of epithelial dysplasia was found exclusively in the non-secretor group.

Laryngorhinootologie, 1999 Jan, 78(1), 47 - 9
{Mycoses of the paranasal sinus system}; Munzel MA; BACKGROUND: During the last few years, in increasing number of fungal infections in the paranasal sinus system has been observed . Aspergillus species as well as mucor and candida albicans are especially responsible for these mycoses . PATIENTS: Twenty-seven cases of a fungal infection of the paranasal sinuses were observed between 1986 and 1997 . The majority of the patients showed a chronic noninvasive form with affection of the maxillary sinus . Other forms (fulminant invasive form, chronic invasive form, allergic fungal sinusitis) are described . Typical features for fungal infection do exist in MRI and CT . CONCLUSIONS: A flexible therapeutic strategy is required in which appropriate pharmacologic and surgical options are tailored to the respective clinical picture.

FEMS Microbiol Lett, 1999 Feb 15, 171(2), 245 - 9
Candida albicans hyphal invasion: thigmotropism or chemotropism?
Davies JM, Stacey AJ, Gilligan CA.
Hyphae of the human pathogenic fungus Candida albicans exhibit thigmotropic behaviour in vitro, in common with phytopathogenic and saprotrophic fungi . An examination of the literature on C . albicans hyphal penetration of epithelial and endothelial membranes does not support the premise that hyphal thigmotropism plays a major role in tissue invasion . Further experimentation is now required to assess thigmotropic behaviour on host membranes and vaginal epithelial cells are suggested as a test model . It is proposed that while thigmotropism may and invasion of tissue invaginations, chemotropism can explain C . albicans hyphal invasion patterns of both endothelium and epithelium.

Am J Obstet Gynecol, 1999 Mar, 180(3 Pt 1), 524 - 9
Vaginal heat shock protein expression in symptom-free women with a history of recurrent vulvovaginitis; Giraldo P et al.; OBJECTIVES: The cause of recurrent vulvovaginitis remains unexplained in most cases . Heat shock protein synthesis is induced under conditions of stress; its presence in vaginal samples from women who were between episodes of recurrent vulvovaginitis thus might reflect a persistent perturbation in the local milieu . Study Design: We undertook an evaluation by means of enzyme-linked immunosorbent assay of 60-kd heat shock protein and inducible 70-kd heat shock protein expressions in vaginal wash samples from 24 symptom-free women with a history of recurrent vulvovaginitis and 19 matched control subjects . The samples were also tested for Candida albicans, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, and human papillomavirus by polymerase chain reaction; for bacterial vaginosis by clinical and microbiologic evaluation; and for interleukin 10, interleukin 1, interleukin 8, RANTES, and eotaxin by enzyme-linked immunosorbent assay . RESULTS: The presence of 60-kd heat shock protein was detected in 11 women with recurrent vulvovaginitis (45.8%) and 1 control subject (5.3%, P =.005) . Similarly, 70-kd heat shock protein was present in 8 patients with recurrent vulvovaginitis (33.3%) and no control subjects (P =.005) . The presence of 60-kd heat shock protein and the presence of 70-kd heat shock protein were correlated with each other (P =.02), as were both 60-kd heat shock protein (P =.006) and 70-kd heat shock protein (P =.01) correlated with IL-10 . There was no relation between the presence of 60-kd heat shock protein or 70-kd heat shock protein and detection of IL-1, IL-8, or any microorganism . CONCLUSION: The expression of heat shock proteins and IL-10 in the vaginas of women with a history of recurrent vulvovaginitis but not in the vaginas of control subjects suggests the existence of differences in the vaginal milieu between the 2 groups, even when both are without vaginal symptoms.

Biochim Biophys Acta, 1999 Feb 2, 1426(3), 409 - 19
Manganese-containing superoxide dismutase and its gene from Candida albicans; Rhie GE et al.; Mitochondrial manganese-containing superoxide dismutase was purified around 112-fold with an overall yield of 1.1% to apparent electrophoretic homogeneity from the dimorphic pathogenic fungus, Candida albicans . The molecular mass of the native enzyme was 106 kDa and the enzyme was composed of four identical subunits with a molecular mass of 26 kDa . The enzyme was not sensitive to either cyanide or hydrogen peroxide . The N-terminal amino acid sequence alignments (up to the 18th residue) showed that the enzyme has high similarity to the other eukaryotic manganese-containing superoxide dismutases . The gene sod2 encoding manganese-containing superoxide dismutase has been cloned using a product obtained from polymerase chain reaction . Sequence analysis of the sod2 predicted a manganese-containing superoxide dismutase that contains 234 amino acid residues with a molecular mass of 26173 Da, and displayed 57% sequence identity to the homologue of Saccharomyces cerevisiae . The deduced N-terminal 34 amino acid residues may serve as a signal peptide for mitochondrial translocation . Several regulatory elements such as stress responsive element and haem activator protein 2/3/4/5 complex binding sites were identified in the promoter region of sod2 . Northern analysis with a probe derived from the cloned sod2 revealed a 0.94-kb band, which corresponds approximately to the expected size of mRNA deduced from sod2.

Med Mycol, 1998 Oct, 36(5), 323 - 30
A dual labelling method for measuring uptake of low molecular weight compounds into the pathogenic yeast Candida albicans; Ziegelbauer K; In contrast to other eukaryotic cells the pathogenic yeast Candida albicans is resistant to many structurally unrelated metabolic inhibitors . Reduced permeability due to the cell wall and/or altered plasma membrane composition is thought to be at least partly responsible for this phenomenon . To study the uptake of low molecular weight compounds into C . albicans we developed a dual labelling method . Intact cells, metabolically inactivated cells, spheroplasts or membrane fragments of C . albicans were incubated with various {14C}-labelled compound in the presence of {3H}-labelled water . After separation of cells and supernatant isotope ratios {3H}/{14C} were determined . Quotients of the isotope ratios from cells and supernatant, called enrichment coefficients, were calculated under all four conditions . The enrichment coefficients indicated whether a compound can enter C . albicans cells, is trapped within the cell wall, is enriched in the lipophilic membrane compartment, is actively accumulated or actively exported by multidrug resistance carriers . We used six structurally unrelated compounds to test our method . We found no evidence for a general impermeability of C . albicans.

Med Mycol, 1998 Oct, 36(5), 269 - 73
Serum interferon-gamma (IFN-gamma) in chronic oral candidosis; Szkaradkiewicz A et al.; Serum IFN-gamma levels were studied in adult patients with chronic oral candidosis, associated with Candida albicans infection . In the group of patients, mean serum IFN-gamma levels (2.74+/-465 pg ml(-1)) were significantly lower than in healthy individuals (9.80+/-1.68 pg ml(-1)) . In analysis of the C . albicans strains isolated from lesions in the patients, their ability was estimated to secrete proteinases . Serum IFN-gamma levels failed to correlate with infections induced by proteinase-producing C . albicans . The results allowed us to conclude that Candida infection may be associated with an insufficient IFN-gamma response of the host, which seems to result in chronic conversion of candidosis . Proteolytic activity of C . albicans strains in vitro is not related to virulence of the strains.

Am J Kidney Dis . 1999 Jan;33(1):E3.
Renal artery rupture secondary to pretransplantation Candida contamination of the graft in two different recipients; Calvino J et al.; Infected graft transplantation is an unwelcome complication that may lead to serious consequences in the immunosuppressed host . It can be caused by infection of the donor or by contamination of the organ during harvest, preservation and handling, or at transplantation . With current donor evaluation protocols, the risk of transmitting infections by exogenous contaminated grafts seems to be more frequent than true donor-transmitted infections . Nevertheless, although rare and usually free of clinically significant sequelae, if contamination is by some virulent organisms such as Staphylococcus aureus, gram-negative bacilli, or fungi, severe complications may occur . We report the clinical outcome of liver, heart, and kidney recipients from a single donor . Both renal allografts had to be removed because of renal artery rupture secondary to Candida albicans infection . Careful donor evaluation before transplantation, unusually early presentation of mycosis leading to anastomotic renal artery disruption, the histopathologic findings of the grafts, and the absence of Candida infection in the liver and heart recipients make us believe that exogenous contamination of the grafts occurred during donor procedure, kidney processing, or at transplantation . In summary, because infected grafts can lead to serious complications, besides careful donor screening, it is important to achieve early recognition of contaminated organs by culturing the perfusate to start specific antibiotic or antifungal therapy after transplantation if necessary and avoid the rare but, in this case, fatal consequences of these infections.

J Clin Microbiol, 1999 Apr, 37(4), 1035 - 44
Development and characterization of complex DNA fingerprinting probes for the infectious yeast Candida dubliniensis; Joly S et al.; Using a strategy to clone large genomic sequences containing repetitive elements from the infectious yeast Candida dubliniensis, the three unrelated sequences Cd1, Cd24, and Cd25, with respective molecular sizes of 15,500, 10,000, and 16,000 bp, were cloned and analyzed for their efficacy as DNA fingerprinting probes . Each generated a complex Southern blot hybridization pattern with endonuclease-digested genomic DNA . Cd1 generated an extremely variable pattern that contained all of the bands of the pattern generated by the repeat element RPS of Candida albicans . We demonstrated that Cd1 does not contain RPS but does contain a repeat element associated with RPS throughout the C . dubliniensis genome . The Cd1 pattern was the least stable over time both in vitro and in vivo and for that reason proved most effective in assessing microevolution . Cd24, which did not exhibit microevolution in vitro, was highly variable in vivo, suggesting in vivo-dependent microevolution . Cd25 was deemed the best probe for broad epidemiological studies, since it was the most stable over time, was the only truly C . dubliniensis-specific probe of the three, generated the most complex pattern, was distributed throughout all C . dubliniensis chromosomes, and separated a worldwide collection of 57 C . dubliniensis isolates into two distinct groups . The presence of a species-specific repetitive element in Cd25 adds weight to the already substantial evidence that C . dubliniensis represents a bona fide species.

J Clin Microbiol, 1999 Apr, 37(4), 925 - 30
Serum is more suitable than whole blood for diagnosis of systemic candidiasis by nested PCR; Bougnoux M et al.; PCR assays for the diagnosis of systemic candidiasis can be performed either on serum or on whole blood, but results obtained with the two kinds of samples have never been formally compared . Thus we designed a nested PCR assay in which five specific inner pairs of primers were used to amplify specific targets on the rRNA genes of Candida albicans, C . tropicalis, C . parapsilosis, C . krusei, and C . glabrata . In vitro, the lower limit of detection of each nested PCR assay was 1 fg of purified DNA from the corresponding Candida species . In rabbits with candidemia of 120 minutes' duration following intravenous (i.v.) injection of 10(8) CFU of C . albicans, the sensitivities of the PCR in serum and whole blood were not significantly different (93 versus 86%) . In other rabbits, injected with only 10(5) CFU of C . albicans, detection of candidemia by culture was possible for only 1 min, whereas DNA could be detected by PCR in whole blood and in serum for 15 and 150 min, respectively . PCR was more often positive in serum than in whole blood in 40 culture-negative samples (27 versus 7%; P < 0.05%) . Lastly, experiments with rabbits injected i.v . with 20 or 200 microgram of purified C . albicans DNA showed that PCRs were positive in serum from 30 to at least 120 min after injection, suggesting that the clearance of free DNA is slow . These results suggest that serum is the sample of choice, which should be used preferentially over whole blood for the diagnosis of systemic candidiasis by PCR.

J Bacteriol, 1999 Mar, 181(6), 1868 - 74
Rapid hypothesis testing with Candida albicans through gene disruption with short homology regions; Wilson RB et al.; Disruption of newly identified genes in the pathogen Candida albicans is a vital step in determination of gene function . Several gene disruption methods described previously employ long regions of homology flanking a selectable marker . Here, we describe disruption of C . albicans genes with PCR products that have 50 to 60 bp of homology to a genomic sequence on each end of a selectable marker . We used the method to disrupt two known genes, ARG5 and ADE2, and two sequences newly identified through the Candida genome project, HRM101 and ENX3 . HRM101 and ENX3 are homologous to genes in the conserved RIM101 (previously called RIM1) and PacC pathways of Saccharomyces cerevisiae and Aspergillus nidulans . We show that three independent hrm101/hrm101 mutants and two independent enx3/enx3 mutants are defective in filamentation on Spider medium . These observations argue that HRM101 and ENX3 sequences are indeed portions of genes and that the respective gene products have related functions.

J Infect Dis, 1999 Apr, 179(4), 1038 - 41
Comparison of interferon-gamma, granulocyte colony-stimulating factor, and granulocyte-macrophage colony-stimulating factor for priming leukocyte-mediated hyphal damage of opportunistic fungal pathogens; Gaviria JM et al.; Proinflammatory cytokines have been proposed as adjunctive therapeutic agents to enhance the host immune response during infections caused by opportunistic fungi . The study compared the differential in vitro priming effects of interferon-gamma (IFN-gamma), granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) on hyphal damage of opportunistic fungi mediated by isolated neutrophils (polymorphonuclear leukocytes, PMNL) and buffy coat cells (polymorphonuclear leukocytes/peripheral blood mononuclear cells, PMNL/PBMC) from healthy donors . IFN-gamma (1000 U/mL) effectively primed both PMNL and PMNL/PBMC for enhanced hyphal damage of Aspergillus fumigatus, Fusarium solani, and Candida albicans . G-CSF (100 ng/mL) increased hyphal damage mediated by both PMNL and PMNL/PBMC against F . solani, and GM-CSF (100 ng/mL) augmented the antifungal activity of PMNL/PBMC against hyphal forms of both F . solani and C . albicans . IFN-gamma may be superior to G-CSF or GM-CSF for enhancing the microbicidal activity of PMNL and PMNL/PBMC against opportunistic fungi.

J Biol Chem, 1999 Mar 12, 274(11), 7286 - 91
The cellular target of histatin 5 on Candida albicans is the energized mitochondrion; Helmerhorst EJ et al.; Histatin 5 is a human basic salivary peptide with strong fungicidal properties in vitro . To elucidate the mechanism of action, the effect of histatin 5 on the viability of Candida albicans cells was studied in relation to its membrane perturbing properties . It was found that both the killing activity and the membrane perturbing activity, studied by the influx of a DNA-specific marker propidium iodide, were inhibited by high salt conditions and by metabolic inhibitors, like sodium azide . In addition, exposure to histatin 5 resulted in a loss of the mitochondrial transmembrane potential in situ, measured by the release of the potential-dependent distributional probe rhodamine 123 . Localization studies using tetramethylrhodamine isothiocyanate-labeled histatin 5 or fluorescein isothiocyanate-labeled histatin 5 showed a granular intracellular distribution of the peptide, which co-localized with mitotracker orange, a permeant mitochondria-specific probe . Like the biological effects, uptake of labeled histatin 5 was inhibited by mitochondrial inhibitors and high salt conditions . Our data indicate that histatin 5 is internalized, and targets to the energized mitochondrion.

Curr Opin Microbiol, 1998 Dec, 1(6), 687 - 92
Dimorphism and virulence in Candida albicans; Mitchell AP; Two regulatory pathways govern filamentation in the pathogenic fungus Candida albicans . Recent virulence studies of filamentation regulatory mutants argue that both yeast and filamentous forms have roles in infection . Filamentation control pathways seem closely related in C . albicans and in Saccharomyces cerevisiae, thus permitting speculation about C . albicans filamentation genes not yet discovered.

Science, 1999 Mar 5, 283(5407), 1535 - 8
Adhesive and mammalian transglutaminase substrate properties of Candida albicans Hwp1; Staab JF et al.; The pathogenesis of candidiasis involves invasion of host tissues by filamentous forms of the opportunistic yeast Candida albicans . Morphology-specific gene products may confer proinvasive properties . A hypha-specific surface protein, Hwp1, with similarities to mammalian small proline-rich proteins was shown to serve as a substrate for mammalian transglutaminases . Candida albicans strains lacking Hwp1 were unable to form stable attachments to human buccal epithelial cells and had a reduced capacity to cause systemic candidiasis in mice . This represents a paradigm for microbial adhesion that implicates essential host enzymes.

Scand J Infect Dis, 1998, 30(5), 527 - 30
Fluconazole therapy in Candida albicans spondylodiscitis; Rossel P et al.; A case of Candida albicans spondylodiscitis in a 20-year-old female liver transplant recipient is reported . The patient was successfully treated with sequential therapy with liposomal amphotericin B and fluconazole . A review of the literature showed 10 cases of Candida albicans spondylodiscitis successfully treated either with fluconazole alone or a sequential therapy with amphotericin B and fluconazole . If long-term amphotericin B therapy is not feasible, a prolonged course of fluconazole in a daily dose of 200-400 mg may be considered as an alternative.

J Med Chem, 1999 Feb 25, 42(4), 584 - 92
Syntheses and biological activities of rebeccamycin analogues . Introduction of a halogenoacetyl substituent; Moreau P et al.; In the course of structure-activity relationships on rebeccamycin analogues, a series of compounds bearing a halogenoacetyl substituent were synthesized with the expectation of increasing the interaction with DNA, possibly via covalent reaction with the double helix . Two rebeccamycin analogues bearing an acetyl instead of a bromoacetyl substituent were prepared to gain an insight into the role of the halogen atom . The new compounds show very little effect on protein kinase C and no covalent reaction with DNA was detected . However, the drugs behave as typical topoisomerase I poisons, and they are significantly more toxic toward P388 leukemia cells than to P388/CPT5 cells resistant to camptothecin . The introduction of a bromo- or chloro-acetyl substituent does not affect the capacity of the drug to interfere with topoisomerase I either in vitro or in cells . One of the bromoacetyl derivatives, compound 8, is the most cytotoxic rebeccamycin derivative among the hundred of derivatives we have synthesized to date . In addition, we determined the antimicrobial activities against two Gram-positive bacteria, Bacillus cereus and Streptomyces chartreusis, and against the Gram-negative bacterium Escherichia coli . The effect of the drugs on Candida albicans yeast growth and their anti-HIV-1 activities were also measured.

J Antimicrob Chemother, 1998 Dec, 42(6), 747 - 53
Synergic effects of tactolimus and azole antifungal agents against azole-resistant Candida albican strains; Maesaki S et al.; We investigated the effects of combining tacrolimus and azole antifungal agents in azole-resistant strains of Candida albicans by comparing the accumulation of {3H}itraconazole . The CDR1-expressing resistant strain C26 accumulated less itraconazole than the CaMDR-expressing resistant strain C40 or the azole-sensitive strain B2630 . A CDR1-expressing Saccharomyces cerevisiae mutant, DSY415, showed a marked reduction in the accumulation of both fluconazole and itraconazole . A CaMDR-expressing S . cerevisiae mutant, DSY416, also showed lower accumulation of fluconazole, but not of itraconazole . The addition of sodium azide, an electron-transport chain inhibitor, increased the intracellular accumulation of itraconazole only in the C26 strain, and not in the C40 or B2630 strains . Addition of tacrolimus, an inhibitor of multidrug resistance proteins, resulted in the highest increase in itraconazole accumulation in the C26 strain . The combination of itraconazole and tacrolimus was synergic in azole-resistant C . albicans strains . In the C26 strain, the MIC of itraconazole decreased from >8 to 0.5 mg/L when combined with tacrolimus . Our results showed that two multidrug resistance phenotypes (encoded by the CDR1 and CaMDR genes) in C . albicans have different substrate specificity for azole antifungal agents and that a combination of tacrolimus and azole antifungal agents is effective against azole-resistant strains of C . albicans.

Mycopathologia, 1998, 142(3), 119 - 23
Ultrastructural effects of date extract on Candida albicans; Shraideh ZA et al.; The effect of Berhi date extract on the ultrastructure of Candida albicans was studied by scanning and transmission electron microscopy . Exposure of yeast to 5% (w/v) date extract showed evidence of weakening in the cell wall with indications of cell distortion and partial collapse in some cases as seen by scanning electron microscopy . Increasing the concentration of date extract (20%, w/v) led to more drastic damage to the yeast with cell lysis and concurrent leakage of cytoplasmic material with eventual cell death . Ultrastructural investigation showed irregular shapes of cells treated with date extract, with prominent effects on cell wall layers . Cell membranes lost their integrity, aggregation of the cytoplasmic contents and large detachment of plasmalemma from cell wall was observed in the treated cells . These results suggest that date extract may have multiple effects on Candida with an increasing potential of using it for prophylaxis purposes.

Antimicrob Agents Chemother, 1999 Mar, 43(3), 702 - 4
Amphotericin B- and fluconazole-resistant Candida spp., Aspergillus fumigatus, and other newly emerging pathogenic fungi are susceptible to basic antifungal peptides; Helmerhorst EJ et al.; The present study shows that a number of basic antifungal peptides, including human salivary histatin 5, a designed histatin analog designated dhvar4, and a peptide from frog skin, PGLa, are active against amphotericin B-resistant Candida albicans, Candida krusei, and Aspergillus fumigatus strains and against a fluconazole-resistant Candida glabrata isolate.

Photochem Photobiol, 1999 Feb, 69(2), 141 - 7
Inhibition of UV-induced immune suppression and interleukin-10 production by plant oligosaccharides and polysaccharides; Strickland FM et al.; Application of Aloe barbadensis poly/oligosaccharides to UV-irradiated skin prevents photosuppression of delayed-type hypersensitivity (DTH) responses in mice . We tested the hypothesis that these carbohydrates belong to a family of biologically active, plant-derived polysaccharides that can regulate responses to injury in animal tissues . C3H mice were exposed to 5 kJ/m2 UVB from unfiltered FS40 sunlamps and treated with between 1 pg and 10 micrograms tamarind xyloglucans or control polysaccharides methylcellulose or dextran in saline . The mice were sensitized 3 days later with Candida albicans . Tamarind xyloglucans and purified Aloe poly/oligosaccharides prevented suppression of DTH responses in vivo and reduced the amount of interleukin (IL)-10 observed in UV-irradiated murine epidermis . Tamarind xyloglucans were immunoprotective at low picogram doses . In contrast, the control polysaccharides methylcellulose and dextran had no effect on immune suppression or cutaneous IL-10 at any dose . Tamarind xyloglucans and Aloe poly/oligosaccharides also prevented suppression of immune responses to alloantigen in mice exposed to 30 kJ/m2 UVB radiation . To assess the effect of the carbohydrates on keratinocytes, murine Pam212 cells were exposed to 300 J/m2 UVB radiation and treated for 1 h with tamarind xyloglucans or Aloe poly/oligosaccharides . Treatment of keratinocytes with immunoprotective carbohydrates reduced IL-10 production by approximately 50% compared with the cells treated with UV radiation alone and completely blocked suppressive activity of the culture supernatants in vivo . The tamarind xyloglucans also blocked UV-activated phosphorylation of SAPK/JNK protein but had no effect on p38 phosphorylation . These results indicate that animals, like plants, may use carbohydrates to regulate responses to environmental stimuli.

Mol Microbiol, 1999 Feb, 31(3), 937 - 47
Selective advantages created by codon ambiguity allowed for the evolution of an alternative genetic code in Candida spp; Santos MA et al.; Several species of the genus Candida decode the standard leucine CUG codon as serine . This and other deviations from the standard genetic code in both nuclear and mitochondrial genomes invalidate the notion that the genetic code is frozen and universal and prompt the questions 'why alternative genetic codes evolved and, more importantly, how can an organism survive a genetic code change?' To address these two questions, we have attempted to reconstruct the early stages of Candida albicans CUG reassignment in the closely related yeast Saccharomyces cerevisiae . These studies suggest that this genetic code change was driven by selection using a molecular mechanism that requires CUG ambiguity . Such codon ambiguity induced a significant decrease in fitness, indicating that CUG reassignment can only be selected if it introduces an evolutionary edge to counteract the negative impact of ambiguity . We have shown that CUG ambiguity induces the expression of a novel set of stress proteins and triggers the general stress response, which, in turn, creates a competitive edge under stress conditions . In addition, CUG ambiguity in S . cerevisiae induces the expression of a number of novel phenotypes that mimic the natural resistance to stress characteristic of C . albicans . The identification of an evolutionary advantage created by CUG ambiguity is the first experimental evidence for a genetic code change driven by selection and suggests a novel role for codon reassignment in the adaptation to new ecological niches.

Yeast, 1999 Jan 30, 15(2), 139 - 43
Over-expression of Candida albicans mitochondrial ribosomal protein S9 (MrpS9p) disturbs mitochondrial function in Saccharomyces cerevisiae; Wiltshire C et al.; A Candida albicans mitochondrial ribosomal protein S9 (MRPS9) cDNA was identified in a screen for sequences whose expression induce galactose lethality in Saccharomyces cerevisiae . MRPS9 appears to encode a protein of 346 amino acids with an N-terminal mitochondrial targeting sequence and an internal S9 signature that is conserved amongst eukaryotic mitochondrial and prokaryotic ribosomal protein S9 sequences . Expression of a GAL1-CaMRPS9 fusion in S . cerevisiae caused the slow development of a galactose-negative phenotype upon repeated subculturing, and this correlated with an increased frequency of petite mutant formation . Therefore, over-expression of CaMRPS9 interferes with S . cerevisiae mitochondrial function, which accounts for the inhibition of growth on galactose.

Yeast, 1999 Jan 30, 15(2), 111 - 21
Asymmetric distribution of phosphatidylethanolamine in C . albicans: possible mediation by CDR1, a multidrug transporter belonging to ATP binding cassette (ABC) superfamily; Dogra S et al.; By using two molecular probes, we demonstrate that only 4% of total phosphatidylethanolamine (PtdEtn) in the plasma membrane (PM) of a human pathogenic yeast, Candida albicans, is present in its external half . Evidence is presented to show that the availability of PtdEtn could be related to the expression of a multidrug transporter CDR1 of C . albicans, and the process is energy-dependent . A homozygous CDR1 disruptant strain of C . albicans shows almost 23% reduction in the external labelling of PtdEtn . This report shows that, similar to human MDRs, yeast multidrug transporter could also be involved in aminophospholipid translocation.

Lett Appl Microbiol, 1999 Jan, 28(1), 7 - 12
An evaluation of the suitability of the European suspension test to reflect in vitro activity of antiseptics against clinically significant organisms; Payne DN et al.; The effectiveness of four antiseptics representing soluble phenolics (Dettol), Quaternary Ammonium Compounds (QAC) (Dettol Hospital Concentrate: DHC), mixed QAC/chlorhexidine (Hibicet Hospital Concentrate: HHC) and povidone iodine (Betadine) was assessed using the proposed phase 2 step 1 European Suspension test . The in vitro activity of the antiseptics against two of the proposed challenge strains, i.e . Staphylococcus aureus and Pseudomonas aeruginosa, was compared with that of 14 problematic clinical isolates of bacteria from a range of genera, including some multiple antibiotic resistant strains, and a clinical isolate of Candida albicans . In addition to the 5 min contact time recommended in the European test, a 1 min time was included . All four products, at their recommended use dilutions and a contact time of 5 min, achieved a Microbicidal Effect (ME) log reduction of at least 5 against the majority of organisms . Differences in activity between products were more pronounced and therefore the tests more discriminatory, when the contact time was reduced to 1 min . The clinical strains were not overtly more resistant to antiseptics than the standard test strains, suggesting that the CEN test strains mimic the antiseptic susceptibility of clinical isolates.

J Am Acad Dermatol, 1999 Feb, 40(2 Pt 2), 322 - 4
Severe paronychia due to zidovudine-induced neutropenia in a neonate; Russo F et al.; We describe the case of an HIV-perinatally exposed child who was treated with zidovudine prophylaxis for reduction of perinatal transmission . At 4 weeks of age, he developed severe paronychia of the great toes as a result of Candida albicans and Escherichia coli . At that time, laboratory tests showed anemia and neutropenia . Zidovudine-related hematologic toxicity resolved after completion of the prophylactic regimen and the infant became HIV-antibody negative (seroreverter) at 8 months of age . Paronychia resolved after treatment with oral fluconazole and topical antiseptics but the soft tissue of the nailfold was penetrated by the edge of the nail plate, resulting in the formation of a cutaneous bridge over the nail that resolved by spontaneous necrosis . To our knowledge, this rare complication has not previously been described in an HIV-perinatally exposed child treated with zidovudine.

Infect Immun, 1999 Mar, 67(3), 1297 - 302
Role of the extracellular signal-regulated protein kinase cascade in human neutrophil killing of Staphylococcus aureus and Candida albicans and in migration; Hii CS et al.; Killing of Staphylococcus aureus and Candida albicans by neutrophils involves adherence of the microorganisms, phagocytosis, and a collaborative action of oxygen reactive species and components of the granules . While a number of intracellular signalling pathways have been proposed to regulate neutrophil responses, the extent to which each pathway contributes to the killing of S . aureus and C . albicans has not been clearly defined . We have therefore examined the effect of blocking one such pathway, the extracellular signal-regulated protein kinase (ERK) cascade, using the specific inhibitor of the mitogen-activated protein kinase/ERK kinase, PD98059, on the ability of human neutrophils to kill S . aureus and C . albicans . Our data demonstrate the presence of ERK2 and a 43-kDa form of ERK but not ERK1 in human neutrophils . Upon stimulation with formyl methionyl leucyl phenylalanine (fMLP), the activities of both ERK2 and the 43-kDa form were stimulated . Despite abrogating the activity of both ERK forms, PD98059 only slightly reduced the ability of neutrophils to kill S . aureus or C . albicans . This is consistent with our finding that PD98059 had no effect on neutrophil adherence or degranulation, although pretreatment of neutrophils with PD98059 inhibited fMLP-stimulated superoxide production by 50%, suggesting that a change in superoxide production per se is not strictly correlated with microbicidal activity . However, fMLP-stimulated chemokinesis was markedly inhibited, while random migration and fMLP-stimulated chemotaxis were partially inhibited, by PD98059 . These data demonstrate, for the first time, that the ERK cascade plays only a minor role in the microbicidal activity of neutrophils and that the ERK cascade is involved primarily in regulating neutrophil migration in response to fMLP.

Infect Immun, 1999 Mar, 67(3), 1063 - 71
Non-serum-dependent chemotactic factors produced by Candida albicans stimulate chemotaxis by binding to the formyl peptide receptor on neutrophils and to an unknown receptor on macrophages; Edens HA et al.; Serum-free culture filtrates of six Candida species and Saccharomyces cerevisiae were found to contain chemoattractants for human polymorphonuclear leukocytes (PMNs) and a mouse macrophage-like cell line, J774 . The chemotactic factors differed for the PMN and J774 cells, however, in terms of heat stability, kinetics of liberation by the yeast cells, and divalent cation requirements for production . The chemoattractant in Candida albicans culture filtrates appeared to act through the formyl peptide receptor (FPR) of PMNs, since it was found to induce chemotaxis of Chinese hamster ovary (CHO) cells that were expressing the human FPR but did not induce chemotaxis of wild-type CHO cells . The C . albicans culture filtrates also induced migration of PMNs across confluent monolayers of a human gastrointestinal epithelial cell line, T84; migration occurred in the basolateral-to-apical direction but not the reverse direction, unless the epithelial tight junctions were disrupted . J774 cells did not migrate toward the formylated peptide (fMet-Leu-Phe; fMLF), and chemotaxis toward the C . albicans culture filtrate was not inhibited by an FPR antagonist (t-butoxycarbonyl-Met-Leu-Phe), suggesting that a different receptor mediated J774 cell chemotaxis . In conclusion, we have identified a receptor by which a non-serum-dependent chemotactic factor (NSCF) produced by C . albicans induced chemotaxis of PMNs . Additionally, we have shown that NSCF was active across epithelial monolayers . These findings suggest that NSCFs produced by C . albicans and other yeast species may influence host-pathogen interactions at the gastrointestinal tract mucosal surface by inducing phagocytic-cell infiltration.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S132 - 8
Evaluation of sexually transmitted diseases diagnostic algorithms among family planning clients in Dar es Salaam, Tanzania; Kapiga SH et al.; OBJECTIVES: To determine the prevalence of sexually transmitted diseases (STDs) and to assess the validity of STD screening approaches among family planning clients in Dar es Salaam, Tanzania . METHODS: Between March and September 1995, information about sociodemographic characteristics, contraceptive use, sexual behaviour, and medical history was obtained from consenting women (n = 908) . After interview, blood and genital specimens were collected for diagnosis of STDs and HIV . Based on the information obtained at interview and clinical examination, STD diagnostic algorithms were developed and validated . RESULTS: The prevalence of STDs was HIV (16.9%), gonococcal and/or chlamydial cervicitis (8.2%), and Trichomonas vaginalis and/or Candida albicans (27.2%) . The risk of cervicitis was increased among unmarried women and among women with a husband < or = 25 years of age and women having more than one sex partners in the past 3 months or a new sex partner during the past month . Most women with cervicitis (62.2%) and vaginitis (67.6%) were asymptomatic . A screening strategy for cervicitis based on symptoms had a sensitivity of 29.7%, a specificity of 84.1%, and a positive predictive value (PPV) of 15.9% . The corresponding figures for an algorithm based on clinical signs were 20.3%, 90.2%, and 15.6% . The sensitivity of a simple risk assessment algorithm ranged from 20.3% to 73% . An approach based on both risk assessment (risk score > or = 1) and clinical signs (cervical mucopus and friability) had a sensitivity of 37.8%, a specificity of 87.5%, and a PPV of 21.4% . A syndromic approach for vaginitis resulted in a higher sensitivity than the approach based on the type of vaginal discharge . CONCLUSION: Although there is no single screening strategy for cervicitis which can be advocated for large scale application, risk assessment might be the only cost effective strategy for identifying women with cervicitis in family planning clinics in Tanzania.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S59 - 76
Risk assessment, symptoms, and signs as predictors of vulvovaginal and cervical infections in an urban US STD clinic: implications for use of STD algorithms; Ryan CA et al.; OBJECTIVE: To identify clinical epidemiological correlates of cervical and vaginal infections and assess alternative algorithms, including two new reproductive tract infection (RTI) algorithms, for syndromic management of these infections . DESIGN, SETTING AND SUBJECTS: We prospectively studied clinical manifestations and risk correlates of cervical and vaginal infections in a randomly sampled group of 779 female patients seeking evaluation for a new problem at a Seattle STD clinic . METHODS: One experienced clinician performed standardised history, physical examination, and microscopy . Reference laboratories performed microbiological tests . Three levels of retrospective evaluation of algorithms included risk assessment and symptom review (RAS) alone; addition of speculum and bimanual examinations; and further addition of microscopy . RESULTS: (1) Chief complaint of abnormal vaginal discharge predicted a significantly lower rate of gonorrhoea (GC) or chlamydial infection (CT) than rates observed with no complaint of vaginal discharge . Only the elicited symptom of yellow vaginal discharge (not the more common symptoms of increased or malodorous vaginal discharge) predicted GC or CT . Chief complaint of abnormal vaginal discharge itself predicted trichomoniasis (TV) and bacterial vaginosis (BV), not cervical infection . Candida albicans was strongly associated with the chief complaint of vulvar pruritus, not with the chief complaint of abnormal vaginal discharge . (2) Applying these algorithms in STD clinics only to women with the chief complaint of abnormal vaginal discharge, rather than to all women, decreases sensitivity for GC or CT, without increasing positive predictive value (PPV) . Criteria for inclusion of patients have more effect on the performance of these algorithms than do the levels of evaluation used . (3) A modified World Health Organisation (WHO) algorithm applied only to patients with symptoms of vaginal discharge, involving treatment of RAS positives for cervical infection, followed by treatment of vaginal infections and cervicitis based on examination of RAS negatives and positives, had a sensitivity of 50% and PPV of 33% for cervical infection, and very low sensitivity for BV, TV, and for vulvovaginal candidiasis (VVC) . (4) An RTI algorithm derived from these data, and applied to all STD patients, involving RAS and examination of all RAS negatives, provided treatment to all cases of BV and TV associated with symptoms of vaginal discharge; treatment of all VVC associated with symptoms of vulvar pruritus; treatment for GC and GT to all RAS positives (using easily elicited risk factors) and to RAS negatives with signs of cervicitis or PID . This algorithm had a sensitivity of 87% and a PPV of 33% for GC or CT in this population, with its 24% prevalence of GC or CT . The sensitivity for BV, TV, and VVC greatly exceeded that of the modified WHO algorithm . (5) A modified RTI algorithm, involving examination rather than treatment of RAS positive women, no examination of RAS negatives, decreased the sensitivity for cervical infection to 55% but increased the PPV to 51% . CONCLUSIONS: Syndromic management of vaginal discharge offers relief of symptoms, prevention of transmission of trichomonas, and perhaps prevention of complications of BV . The 51% PPV of the modified RTI algorithm probably would warrant treatment and partner notification for GC and CT in settings with similar rates of GC and CT where more specific tests are lacking . However, as the prevalence of GC or CT decreases, the ratio of uninfected to infected who receive treatment with these algorithms would increase greatly, making the algorithms potential victims of their own success.

Pediatr Res, 1999 Feb, 45(2), 224 - 9
Differential effects of glucocorticoids on colony stimulating factors produced by neonatal mononuclear cells; Witek-Janusek L et al.; The purpose of this study was to examine the effect of dexamethasone (DEX) on the production of granulocyte and granulocyte-macrophage colony stimulating factors (G-CSF and GM-CSF) by neonatal mononuclear cells . Mononuclear cells were isolated from umbilical cord blood and cultured with either phorbol myristate acetate/phytohemagglutinin (PMA/PHA) or Candida albicans with or without DEX (10(-8)-10(-6) M) for 48 h . Cell supernatants were assayed for G-CSF and GM-CSF by ELISA . Mononuclear cells from term and preterm infants responded to PMA/PHA stimulation with a significant increase in G-CSF production over baseline levels . The PMA/PHA-induced increase in G-CSF production was markedly augmented by the addition of DEX to cell cultures . DEX augmented production of G-CSF was significantly less in mononuclear cells from preterm infants . Similarly, production of G-CSF was significantly less by mononuclear cells from infants with acute physiology scores of > or = 10, as judged by the Score for Acute Neonatal Physiology . In contrast, DEX significantly inhibited PMA/PHA-induced GM-CSF production . Although C . albicans induced mononuclear cells to produce G-CSF, DEX did not significantly augment this production . No significant effect of DEX on C . albicans induced GM-CSF production was observed . The data show DEX induced differential regulation of infant peripheral blood mononuclear cell production of G-CSF and GM-CSF . These results suggest that glucocorticoids may enhance certain aspects of host immune function in addition to their well-documented immunosuppressive effects . Further, the neutrophilia observed in DEX-treated infants may be due to enhanced G-CSF production.

J Med Microbiol, 1999 Feb, 48(2), 167 - 72
Scanning electron microscopy characterisation of colonies of Candida albicans morphological mutants; Pesti M et al.; The ultrastructures of colonies of two stable UV-induced morphological mutants and their parental strain of Candida albicans grown on glucose-containing solid medium were investigated by scanning electron microscopy . The structures and ultrastructures of these three types of colonies were determined not only in terms of the proportions of blastospores, hyphae and pseudohyphae, but also with regard to the mode of budding of blastospores and the positions of these particular cell types within the colonies . Hyphae with an atypical appearance and branching characters were observed both in regular-wrinkled and in irregular-wrinkled mutant colonies . Smooth colonies of the parental strain and the mutants exhibited the same hyphal network within the agar, suggesting that micro-environmental factors in the agar overcame the effects of these mutations.

Med Mycol, 1998, 36 Suppl 1, 238 - 48
Molecular mechanisms of virulence in fungus-host interactions for Aspergillus fumigatus and Candida albicans; Muhlschlegal F et al.; Research on fungi that cause opportunistic infections has increased dramatically during the past few years, largely because these organisms cause significant morbidity and mortality . Most of this research has focused on defining the virulence factors produced by these pathogens, as well as developing methods for the diagnosis of fungal diseases . With regard to studies on the biology of Candida albicans, it is now possible to isolate genes, disrupt their expression, and observe the specific effects of gene disruption on virulence and growth of the organism . Moreover, growth and virulence of this pathogen is also being studied and the effect of environmental factors on gene expression investigated . This subject is especially important in view of the fact that C . albicans can colonize and invade a number of sites in the human body . Thus, its ability to grown in the oral and vaginal tracts, as well as in blood, requires the organism to adapt to a variety of environmental stresses . Here we present observations on the growth, morphogenesis and virulence of the opportunistic fungi C . albicans and Aspergillus fumigatus.

Med Mycol, 1998, 36 Suppl 1, 230 - 7
Advances in molecular genetics of Candida albicans and Candida glabrata; Brown AJ et al.; Ten years ago, when molecular genetic methods were being applied vigorously to viruses, bacterial pathogens and eukaryotic parasites, there seemed to be a partial paralysis in applying them to infectious fungi; this state of affairs was more than apparent in the composition of the symposia at the ISHAM conference in 1987 . Since then, however, things have changed . The ISHAM conference held in Italy in 1997 was replete with studies utilizing molecular genetic techniques to answer questions related to epidemiology, pathogenesis, drug development and typing . In the symposium Advances in Molecular Genetics of Fungal Pathogens, several new applications of molecular biology to fungal pathogenesis were reviewed . Although the presentations in this symposium covered only a fraction of the molecular methods now being applied to Candida pathogenesis, they nevertheless provided an intriguing view of what is in store for us in the coming years.

Med Mycol, 1998, 36 Suppl 1, 156 - 65
Importance of Candida species other than Candida albicans as opportunistic pathogens; Coleman DC et al.; Candida species other than C . albicans have become a significant cause of infection in humans . Several of the more commonly isolated of these species are less susceptible to commonly used azole antifungal drugs, a factor that poses significant difficulties for effective treatment . The modern mycology laboratory has an important role to play in several aspects relating to these organisms, including therapy, detection, identification and epidemiological analysis . The application of molecular techniques and phylogenetic analysis has led to the identification of a new species of Candida associated with mucosal candidiasis in HIV-infected individuals named Candida dubliniensis, the clinical significance of which is currently under investigation . Molecular techniques are also being applied to the analysis of determinants involved in pathogenicity of species such as Candida glabratta . These approaches should lead to a better understanding of these organisms and there ability to cause disease and should also provide more effective treatment.

Med Mycol, 1998, 36 Suppl 1, 95 - 105
Antibody and/or cell-mediated immunity, protective mechanisms in fungal disease: an ongoing dilemma or an unnecessary dispute?
Casadevall A, Cassone A, Bistoni F, Cutler JE, Magliani W, Murphy JW, Polonelli L, Romani L.
Historically there has been controversy on the relative importance of antibody- and cell-mediated immune responses in the protection against fungal pathogens . The controversy was fuelled by the difficulties encountered in obtaining consistent results with polyclonal antibody experiments and I inducing long-lasting immune protection by vaccinations with induce stron cell-mediated responses . Recent studies indicate that both antibody- and cell-mediated immune responses can contribute to host protection against Candida albicans and C . neoformans . At the present time the major issue is not the relative importance of antibody- and cell-mediated immune responses but rather, the mechanisms by which the two arms of the immune system function and cooperate.

J Bacteriol, 1999 Feb, 181(4), 1369 - 73
Lack of genetic differentiation between two geographically diverse samples of Candida albicans isolated from patients infected with human immunodeficiency virus; Xu J et al.; The patterns of genetic variation of samples of Candida albicans isolated from patients infected with human immunodeficiency virus in Durham, N.C., and Vitoria, Brazil, were compared . Genotypes for 126 strains were obtained at 16 polymorphic restriction sites distributed on nine PCR fragments . The results indicated evidence of clonality both within and between these two geographically diverse samples . The samples are genetically very similar, with little evidence of genetic differentiation.

J Infect Dis, 1999 Mar, 179(3), 661 - 9
Effective phagocytosis and killing of Candida albicans via targeting FcgammaRI (CD64) or FcalphaRI (CD89) on neutrophils; van Spriel AB et al.; Invasive fungal infections are an increasing problem for immunocompromised patients . As an approach to improve targeting of polymorphonuclear leukocytes (PMNL) toward Candida albicans, the effect of bispecific antibodies (BsAbs) directed against C . albicans and either FcalphaRI or FcgammaRI was evaluated . Control PMNL and in vivo granulocyte colony-stimulating factor (G-CSF)-primed PMNL served as effector cells . A new radiometric killing assay for measuring candidacidal activity was developed to facilitate quantification of PMNL-mediated killing of C . albicans . BsAbs directed to either FcgammaRI (CD64) or FcalphaRI (CD89) on human PMNL effectively enhanced both phagocytosis and killing of C . albicans in vitro . Fungicidal activity triggered via FcgammaRI required in vivo priming with G-CSF, whereas FcalphaRI-mediated activity was not dependent on this growth factor . Furthermore, PMNL from human FcgammaRI-transgenic mice effectively phagocytosed and eliminated C . albicans in the presence of BsAbs . These results document the capacity of FcR-directed BsAbs and G-CSF to trigger antifungal immune responses.

FEMS Microbiol Lett, 1999 Jan 15, 170(2), 319 - 25
2,4-(hydroxyphenyl)-ethanol, an antioxidative agent produced by Candida spp., impairs neutrophilic yeast killing in vitro; Cremer J et al.; Culture supernatants of Candida albicans were examined for factors with inhibitory activity against the chemiluminescence of human neutrophils . By high resolution gel chromatography, a low-molecular-mass chemiluminescence inhibitor was isolated . The compound was identified as 2,4-(hydroxyphenyl)-ethanol . Half-maximum inhibition (IC50) of the chemiluminescence response of neutrophils phagocytizing opsonized zymosan or C . albicans occurred at 38.1 +/- 2.3 microM and 19.9 +/- 8.3 microM, respectively . As shown by flow cytometry, the compound protected C . albicans against phagocytic killing (IC50 = 73.8 +/- 16.9 microM) . Substantially higher concentrations of the inhibitor were produced by C . albicans and C . tropicalis than by C . parapsilosis and C . glabrata, suggesting a potential role in pathogenicity ranking.

J Biol Chem, 1999 Feb 12, 274(7), 4000 - 8
Oligomeric structure and regulation of Candida albicans glucosamine-6-phosphate synthase; Milewski S et al.; Candida albicans glucosamine-6-phosphate (GlcN-6-P) synthase was purified to apparent homogeneity with 52% yield from recombinant yeast YRSC-65 cells efficiently overexpressing the GFA1 gene . The pure enzyme exhibited Km(Gln) = 1.56 mM and Km(Fru-6-P) = 1.41 mM and catalyzed GlcN-6-P formation with kcat = 1150 min-1 . The isoelectric point of 4.6 +/- 0.05 was estimated from isoelectric chromatofocusing . Gel filtration, native polyacrylamide gel electrophoresis, subunit cross-linking, and SDS-polyacrylamide gel electrophoresis showed that the native enzyme was a homotetramer of 79.5-kDa subunits, with an apparent molecular mass of 330-340 kDa . Results of chemical modification of the enzyme by group-specific reagents established an essential role of a cysteinyl residue at the glutamine-binding site and histidyl, lysyl, arginyl, and tyrosyl moieties at the Fru-6-P-binding site . GlcN-6-P synthase in crude extract was effectively inhibited by UDP-GlcNAc (IC50 = 0.67 mM) . Purification of the enzyme markedly decreased the sensitivity to the inhibitor, but this could be restored by addition of another effector, glucose 6-phosphate . Binding of UDP-GlcNAc to the pure enzyme in the presence of Glc-6-P showed strong negative cooperativity, with nH = 0.54, whereas in the absence of this sugar phosphate no cooperative effect was observed . Pure enzyme was a substrate for cAMP-dependent protein kinase, the action of which led to the substantial increase of GlcN-6-P synthase activity, correlated with an extent of protein phosphorylation . The maximal level of activity was observed for the enzyme molecules containing 1 . 21 +/- 0.08 mol of phosphate/mol of GlcN-6-P synthase . Monitoring of GlcN-6-P synthase activity and its sensitivity to UDP-GlcNAc during yeast --> mycelia transformation of C . albicans cells, under in situ conditions, revealed a marked increase of the former and a substantial fall of the latter.

Mycopathologia, 1998, 142(2), 67 - 70
Cloning and sequence analysis of Histoplasma capsulatum glucosamine-6-phosphate synthase gene fragment; Sachadyn P et al.; The 3' part of the glucosamine-6-phosphate synthase gene from Histoplasma capsulatum was PCR amplified using degenerate primers designed from the known glucosamine-6-phosphate synthase gene sequences, cloned and sequenced . The computer analysis of the 676 bp sequence revealed the presence of two introns . The identities of the deduced amino acid sequence to the corresponding Saccharomyces cerevisiae and Candida albicans fragment are 65 and 63.8%, respectively.

FEBS Lett, 1999 Jan 8, 442(1), 53 - 6
Isolation of p-hydroxycinnamaldehyde as an antibacterial substance from the saw fly, Acantholyda parki S; Leem JY et al.; We purified an antibacterial substance from larvae of the saw fly, Acantholyda parki S., and identified its molecular structure as p-hydroxycinnamaldehyde . We then synthesized it by reduction of p-hydroxycinnamic acid . The antibacterial activity of the synthetic p-hydroxycinnamaldehyde was equal to that of the authentic substance . This molecule was found to have a broad antibacterial spectrum against not only Gram-negative, but also Gram-positive bacteria . Furthermore, it showed antifungal activity against Candida albicans . We suggest that this substance may play a role in the defense system of this insect . This is the first report of p-hydroxycinnamaldehyde of animal origin.

J Prosthet Dent, 1999 Feb, 81(2), 207 - 14
Microwave disinfection of denture base materials colonized with Candida albicans; Dixon DL et al.; STATEMENT OF PROBLEM: Infection of denture materials with Candida albicans is common and contributes to denture stomatitis . PURPOSE: This 3-phase investigation examined: (1) the efficacy of microwave irradiation against C albicans colonized on 3 soft denture liners and 1 heat-polymerized denture base resin, and (2) the effect of this irradiation on the hardness of the materials tested . MATERIAL AND METHODS: In phase 1, an experimental protocol was developed . Sterilized specimens from 2 denture base soft liners and 1 heat-polymerized acrylic resin denture base material (n = 45 each) were inoculated with C albicans . Two thirds of the specimens were irradiated in a 60 Hz microwave oven for 5 minutes (dry) . C albicans growth was then assessed with streaked blood agar plates and thioglycollate broth . One third of the specimens were not irradiated and served as controls . Pretest and posttest Shore A hardness values were obtained and compared . For phase 2, 15 specimens from each material group were subjected to irradiation (while immersed in water) for 5 minutes; and, 15 from each material were subjected to 10- and 15-minute irradiation (dry), with subsequent sterility and change in hardness assessments completed as described in phase 1 . In phase 3, 15 specimens from each material group were subjected to repeated 5-minute irradiation cycles (while immersed in water), and changes in hardness were examined . RESULTS: Only the 5-minute irradiated specimens immersed in water were effectively sterilized, as verified by the thioglycollate assay . The effect of repeated 5-minute irradiation cycles resulted in a significant change in hardness of the PermaSoft specimens . CONCLUSIONS: Five-minute irradiation, while immersed in water, killed all C albicans present on the materials tested; and, repeated 5-minute irradiation significantly affected the hardness of only the PermaSoft material.

J Bacteriol, 1999 Feb, 181(3), 700 - 8
The bZip transcription factor Cap1p is involved in multidrug resistance and oxidative stress response in Candida albicans; Alarco AM et al.; Candida albicans is an opportunistic pathogenic yeast which frequently develops resistance to the antifungal agent fluconazole (FCZ) in patients undergoing long-term therapy . FCZ-resistant strains often display a reduced intracellular FCZ accumulation which correlates with the overexpression of the ATP-binding cassette transporters CDR1 and CDR2 or the major facilitator (MF) MDR1 . We have recently cloned a C . albicans gene, named CAP1, which codes for a bZip transcription factor of the AP-1 family homologous to the Yap1 protein involved in multidrug resistance and response to oxidative stress in Saccharomyces cerevisiae . CAP1 was found to confer FCZ resistance in S . cerevisiae by transcriptionally activating FLR1, a gene coding for an MF homologous to the C . albicans MDR1 gene product (A.-M . Alarco, I . Balan, D . Talibi, N . Mainville, and M . Raymond, J . Biol . Chem . 272:19304-19313, 1997) . To study the role of CAP1 in C . albicans, we constructed a CAI4-derived mutant strain carrying a homozygous deletion of the CAP1 gene (CJD21) . We found that deletion of CAP1 did not affect the susceptibility of CJD21 cells to FCZ, cerulenin, brefeldin A, and diamide but caused hypersensitivity to cadmium, 4-nitroquinoline N-oxide, 1,10-phenanthroline, and hydrogen peroxide, an effect which was reverted by reintroduction of the CAP1 gene in these cells . Introduction of a hyperactive truncated allele of CAP1 (CAP1-TR) in CJD21 resulted in resistance of the cells to all of the above compounds except hydrogen peroxide . The hyperresistant phenotype displayed by the CJD21 CAP1-TR transformants was found to correlate with the overexpression of a number of potential CAP1 transcriptional targets such as MDR1, CaYCF1, CaGLR1, and CaTRR1 . Taken together, our results demonstrate that CAP1 is involved in multidrug resistance and oxidative stress response in C . albicans . Finally, disruption of CAP1 in strain FR2, selected in vitro for FCZ resistance and constitutively overexpressing MDR1, did not suppress but rather increased the levels of MDR1 expression, demonstrating that CAP1 acts as a negative transcriptional regulator of the MDR1 gene in FR2 and is not responsible for MDR1 overexpression in this strain.

Yakugaku Zasshi, 1998 Dec, 118(12), 616 - 20
{Inhibition of mycelial growth of Candida albicans by ascites fluid of tumor bearing mice}; Ubukata T et al.; The effects of ascites fluids and sera of tumor-bearing mice on the mycelial growth of Candida albicans were examined . When the ascites fluids or the sera obtained from mice inoculated with MM46 mammary carcinoma were added to the culture medium, mycelial growth of C . albicans was strongly inhibited . The molecular size of the growth inhibitory factor in the ascites fluids was estimated to be approximately 80 K dalton by gel-filtration chromatography . Ferric chloride (6 microM) neutralized the anti-Candida activity . On the basis of these results including morphological observation, a possible role of a transferrin-like molecule was discussed.

Mycoses, 1998 Dec, 41(11-12), 487 - 92
Comparison of fungal viability assays using Candida albicans yeast cells undergoing prolonged incubation in the absence of nutrients; Hjertstedt J et al.; Staining methods for determining fungal viability are usually assessed by comparisons with enumeration of colony-forming units (CFU) on solid media . The purpose of the present study was to compare viability as assessed by the acridine orange (AO) and MTT methods with the numbers of CFUs obtained for Candida albicans yeast cells undergoing prolonged incubation in distilled water . In initial assessments of the assays using various proportions of control and heat-killed C . albicans, the AO and MTT methods consistently indicated significantly higher values for viability than did CFU determinations . Experiments using organisms cultured overnight revealed that approximately 95% of the cells were capable of dividing at least once in a microscopic proliferation assay, whereas only 69% were capable of forming colonies . Parallel assays comparing AO uptake and MTT reduction gave excellent agreement with the microscopic proliferation assay, but not with CFU determinations . Using organisms undergoing prolonged incubations in distilled water, much lower viabilities were obtained with the CFU method at 7 and 10 days than with the microscopic proliferation assay or the two staining methods . These results indicate that the AO and MTT assays correlate well with the ability of C . albicans to divide at least once, but may not accurately indicate the percentage of organisms actually able to form colonies.

Mycoses, 1998 Dec, 41(11-12), 481 - 6
Imidazole-induced morphological abnormalities of mitochondria of Candida albicans; Yang HC et al.; The mitochondrial morphology of live Candida albicans samples treated with several imidazoles (miconazole, econazole and clotrimazole) was observed with a fluorescent carbocyanine probe, 3,3'-dihexyloxacarbocyanine {diO-C6-(3)}, under a fluorescence microscope . Nearly all non-treated C . albicans cells carried only long tubular mitochondria . Treatment with antimycotics at half the minimum inhibitory concentrations (MIC100) rapidly induced mitochondrial cleavage or fragmentation, which was followed by recovery to the normal tubular morphology within 1 h . Exposing the yeast to drugs at concentrations higher than the MICs resulted in the development of swollen mitochondria or amorphous bodies . These phenomena were concentration dependent . The fluorescence images were also compared with ultrastructural images obtained by transmission electron microscopy.

Mycoses, 1998 Dec, 41(11-12), 477 - 80
Colorimetric MTT assessment of antifungal activity of D0870 against fluconazole-resistant Candida albicans; Yang HC et al.; The in vitro antifungal activity of D0870 against eight isolates of fluconazole-resistant Candida albicans was compared with that of itraconazole, ketoconazole and miconazole . The colorimetric MTT {3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide} assay was used to assess the antifungal activities . The 50% minimum inhibitory concentration (MIC50) of D0870 was below 0.031 microgram ml-1 for seven isolates and 0.25 microgram ml-1 for one isolate . The activity of D0870 was superior to that of the other azoles . Ketoconazole was the most effective azole next to D0870 . Therefore, the new bis-triazole, D0870, is expected to be promising for the therapy of fluconazole-resistant candidosis . The present data also confirmed that the MTT assay may be useful for evaluation of resistance and detection of resistant C . albicans.

Mol Ecol, 1999 Jan, 8(1), 59 - 73
PCR-restriction fragment length polymorphism (RFLP) analyses reveal both extensive clonality and local genetic differences in Candida albicans; Xu J et al.; Using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method to obtain genotypes for the diploid pathogenic yeast, Candida albicans, we analysed 204 C . albicans isolates from three populations of the Duke University community: two from clinical sources {one from patients infected with human immunodeficiency virus (HIV) and the other from patients without HIV infection}, and the third from healthy student volunteers . The results indicated: (i) extensive evidence for clonality within and between populations of C . albicans; and (ii) greater genotypic and gene diversities in the nonclinical population than those derived from clinical specimens, regardless of HIV status . The two clinical populations were genetically more similar to each other than either was to the population consisting of isolates from healthy people . Within each population sample there was a general lack of heterozygotes, and random associations of alleles within and between loci were found in less than 50% of the loci or pairs of loci . These findings were consistent between the two sets of samples analysed: those including all isolates and those including only clone-corrected isolates . Possible mechanisms are presented to explain the observed patterns of genetic variation within and between C . albicans populations.

Mycoses, 1998 Nov, 41(9-10), 421 - 3
Fluconazole prophylaxis in patients with head and neck tumours undergoing radiation and radiochemotherapy; Mucke R et al.; The aim of the present study was to investigate the incidence of Candida stomatitis and resulting interruptions in radiation and radiochemotherapy in 50 patients suffering from squamous cell carcinomas of the head and neck region receiving fluconazole (100 mg d-1) in comparison with a historical control group (n = 50) without specific prophylaxis . Twenty of the control patients (40%) demonstrated Candida stomatitis, with seven of them (14%) requiring interruptions in anticancer therapy . In contrast, none of the patients with fluconazole had evidence of Candida stomatitis (P = 0.0000051) and subsequent interruption of anti-cancer therapy (P = 0.0061) . Laboratory monitoring for the presence of Candida species was performed in 30 patients before and after therapy with fluconazole . Candida albicans was identified less frequently after therapy when compared with the pretreatment status . However, C . glabrata and C . krusei were isolated in some of the patients, probably because of drug resistance of these subspecies . The results demonstrate the clinical usefulness of prophylactic fluconazole applications in patients suffering from head and neck tumours with the aim of reducing Candida stomatitis and the resulting interruptions in radiation and radiochemotherapy.

Mycoses, 1998 Nov, 41(9-10), 411 - 6
Study of the action of alpha-hederin on the ultrastructure of Candida albicans; Moulin-Traffort J et al.; alpha-Hederin, a saponin isolated from Hedera helix (L.) (Araliaceae), was tested on Candida albicans ultrastructure . The concentrations used were 6.25, 12.5, and 25 micrograms ml-1 for an exposure time of 24 h . Transmission electron microscopy observations indicated that compared with untreated control yeasts, alpha-hederin induced modifications of cellular contents and alterations of cell envelope with degradation and death of the yeasts . The impact of alpha-hederin on the biomembranes and in particular on the plasmalemma is discussed . The antifungal activity of alpha-hederin was confirmed, as was the minimal inhibitory concentration (25 micrograms ml-1).

Mycoses, 1998 Nov, 41(9-10), 397 - 402
Typing of Candida albicans and Candida parapsilosis: species-related limitations of electrophoretic karyotyping and restriction endonuclease analysis of genomic DNA; Riederer K et al.; Species-related discrimination limits of electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG), using pulse-field gel electrophoresis, in typing Candida albicans (CA) and Candida parapsilosis (CP) were compared . Eleven CA and 12 CP isolates from individual neonates and three CA and three CP control isolates were used . For CA, EK and REAG with sfiI displayed seven and six banding-patterns, respectively . One karyotype and two SfiI banding-patterns were observed among the control-isolates . Combining EK/REAG (SfiI) demonstrated nine composites and three distinct control-composites . For CP, EK displayed nine karyotypes, REAG (SfiI) demonstrated four banding-patterns, and REAG (BssHII) yielded six banding-patterns . EK and REAG/SfiI failed to distinguish any CP-controls whereas REAG/BssHII distinguished 2/3 CP-controls . Combining EK/REAG (SfiI) showed 10 composites indistinguishable from CP-controls whereas EK/REAG (BssHII) demonstrated 11 composites and three distinct control-composites . These results illustrate that singly, EK and REAG have significant limitation in typing Candida species though EK is more precise . Combining both methods yields better results but the appropriate restriction endonuclease may vary by strains or species . These findings underscore the importance of combining multiple typing methods, testing several control isolates, and correlating the results with careful epidemiological assessment.

Kansenshogaku Zasshi, 1998 Dec, 72(12), 1306 - 10
{A case with chronic active EB virus infection accompanied with pulmonary candidiasis}; Karino T et al.; A 44-year-old woman with a history of intermittent fever for several years was admitted because of burn on her leg . On admission, she had hepatosplenomegaly and fever . Antibiotic therapy was started for bacterial infection of the burn . She lost her appetite and IVH was started . During the treatment, high fever appeared and chest X-ray films showed multiple nodular infiltrates throughout both lung fields . Candida albicans was isolated from IVH catheter culture and pulmonary candidiasis was suspected . Her fever and lung involvements were successfully treated with fluconazole . During the course, serum anti-EB-VCA-IgG antibody persisted at a high titer and anti-EBNA antibody remained negative . EB virus DNA was detected in the peripheral blood and bone marrow . Thus, she was diagnosed as chronic active EB virus infection.

Zentralbl Hyg Umweltmed, 1998 Dec, 201(4-5), 337 - 47
{Effect of steam application based on microbiological and parasitologic test procedures}; Haas A et al.; In the present study steam application was investigated with regard to microbicidal and parasiticidal effects . The cleaning apparatus used (Uninova Company) works at a boiler pressure of about 5 bar and consequently with a temperature up to 155 degrees C inside the boiler . Whereas the ambient atmosphere working temperature of steam is slightly below 100 degrees C . The tests are based on the DVG guidelines for testing chemical disinfectants (2) . Different steaming times and distances were used in germ carrier tests with three different germ carriers (tile, wood, carpet) and three different test germs (Staphyloccocus aureus, Pseudomonas aeruginosa, Candida albicans) in order to determine the optimum conditions for biocidal effects of steam-application . These optimum conditions were additionally tested with two test viruses (ECBO- and Reo-virus) and a parasitological resting form (ascarid worm eggs) . Swirling of germs caused by steam turbulence was minimized by covering the steam outlet nozzle with cloth . The experiments showed logarithmical reduction factors of at least 5.0 in the germ count at steaming times of 5 seconds and a steaming distance of 2.5 cm for all three test germs on all three germ carriers (mean of 10 repeated tests) . The virological tests showed good disinfection results after a steaming time of only 2 seconds using aseptic gauze as germ carrier and also after 5 seconds using wood as a carrier . Finally in testing vitality of undeveloped Ascarid worm eggs only 2 seconds of steam treatment proved to be sufficient for a 100 percent destruction . According to the present results steam treatment is most likely to become a valuable, ecologically compatible method in controlling hygienic problems, with a potential of partly replacing chemical disinfectants . In particular we see applications in keeping pets and companion animals, provided the above mentioned rules are followed (steaming distance 2.5 cm; steaming time 5 seconds; cloth) . In farm animal stables steam disinfection seems harder to achieve because of large, rough surfaces and economical reasons as e.g . expenditure of time and energy.

Infect Immun, 1999 Feb, 67(2), 826 - 33
Effectiveness of a vaccine composed of heat-killed Candida albicans and a novel mucosal adjuvant, LT(R192G), against systemic candidiasis; Cardenas-Freytag L et al.; The incidence of fungal infections caused by the opportunistic yeast Candida albicans has increased significantly in recent years . The ability to vaccinate selected patients against the organism would be advantageous . In this paper we describe a potential anti-C . albicans vaccine consisting of heat-killed C . albicans (HK-CA) in combination with the novel mucosal adjuvant LT(R192G), a genetically detoxified form of the heat-labile toxin of enterotoxigenic Escherichia coli . Groups of male CBA/J mice were immunized intranasally on three occasions at weekly intervals with 2 x 10(7) HK-CA per dose, alone or in conjunction with 10 micrograms of LT(R192G) per dose . Two weeks following the last application of antigen, some animals were challenged intravenously (i.v.) with 10(4), 10(5), or 10(6) viable C . albicans to assess protection as measured by survival and/or culture . Some groups of animals were footpad tested with C . albicans mannan to assess delayed-type hypersensitivity (DTH), and all the animals were bled for antibody assays . In two independent studies, all the animals immunized with HK-CA plus LT(R192G) were able to eradicate 10(4) C . albicans completely, as determined by kidney culture 4 weeks after challenge . Animals immunized with HK-CA only had reduced levels of C . albicans compared to the adjuvant or saline-only control . Greatly enhanced survival was observed when mice immunized with HK-CA plus LT(R192G) were challenged with 10(5) live C . albicans as well . Animals immunized with HK-CA plus LT(R192G) developed a significant DH response, while those given HK-CA alone developed only marginal DH responses . High immunoglobulin G (IgG) levels to cytoplasmic antigens developed in mice immunized with HK-CA plus LT(R192G), but they were found only after i.v . challenge . Addition of adjuvant shifted the antibody isotype production in i.v.-challenged animals to a response dominated by IgG2a . Clearly, intranasal immunization with killed C . albicans in conjunction with LT(R192G) afforded significant levels of protection . This novel approach offers new possibilities for the development of an effective vaccine against candidiasis for use in humans.

Infect Immun, 1999 Feb, 67(2), 670 - 4
Early resistance of interleukin-10 knockout mice to acute systemic candidiasis; Vazquez-Torres A et al.; In contrast to immunocompetent controls, interleukin-10 (IL-10) knockout (KO) mice eliminated an experimental intravenous inoculation with Candida albicans from their kidneys . Improved clearance of C . albicans from the kidneys of IL-10 KO mice was evident at 24 h after intravenous challenge with the fungus . Conversely, mice with a deletion of the IL-4 cytokine gene were more susceptible to systemic candidiasis than were immunocompetent controls . The hyperresistance of IL-10 KO mice to acute systemic candidiasis did not seem to correlate with nitric oxide-mediated immunity, but rather, it appeared to be associated with more efficient effector function of innate cells, possibly neutrophils . In support of the latter hypothesis, we observed that neutrophils from IL-10 KO mice were more efficient at killing C . albicans blastoconidia and hyphae than were neutrophils from immunocompetent control mice . Neither IL-10 KO nor IL-4 KO mice that were monoassociated with C . albicans for 4 weeks showed any histologic evidence of systemic candidiasis of endogenous origin . In contrast to systemic candidiasis, we observed no significant (P < 0.05) differences in susceptibility among IL-10 KO, IL-4 KO, and wild-type (immunocompetent) mice to orogastric candidiasis . Our results suggest that IL-10 exerts a negative effect on the early, innate response to acute systemic candidiasis; however, in comparison to immunocompetent control (wild-type) mice, neither IL-10 nor IL-4 deficiency enhanced susceptibility to orogastric candidiasis.

Microb Pathog, 1998 Dec, 25(6), 349 - 52
A second Candida albicans resistance gene (Carg2) regulates tissue damage, but not fungal clearance, in sub-lethal murine systemic infection; Ashman RB et al.; The severity of systemic infection with the yeast Candida albicans has been shown to be under complex genetic control . C57/L mice carry an allele that is associated with an increase in tissue destruction when compared with C57Bl/6 mice; however, the gene affects only the severity of tissue lesions, and does not influence the magnitude of the fungal burden in either kidney or brain . Studies in {C57/LxC57Bl/6}F1 hybrid mice, and {C57/LxC57Bl/6}F1xC57/L backcross mice, demonstrated that the gene behaves as a simple Mendelian co-dominant .

Microb Pathog, 1998 Dec, 25(6), 333 - 5
A gene (Carg1) that regulates tissue resistance to Candida albicans maps to chromosome 14 of the mouse; Ashman RB; Tissue susceptibility and resistance to infection with the yeast Candida albicans is genetically regulated . Analysis of the strain distribution pattern of the C . albicans resistance gene (Carg1) and additional gene and DNA segment markers in the AKXL recombinant inbred (RI) set showed that 13/15 RI strains were concordant for Carg1, Tcra and Rib1 . Therefore, Carg1 is probably located within a 17 cM segment of chromosome 14, within approximately 4 cM of the other two genes .

Mol Cells, 1998 Dec 31, 8(6), 786 - 9
Bacterial expression of tenecin 3, an insect antifungal protein isolated from Tenebrio molitor, and its efficient purification; Kim DH et al.; Tenecin 3, an antifungal protein isolated from the insect Tenebrio molitor larvae, inhibits growth of the fungus Candida albicans . However, the antifungal mechanism and functions of tenecin 3 have not yet been studied due to its very low availability from the natural source . Here we report an expression system of the recombinant tenecin 3 in E . coli, whose amino acid composition is the same with that of the natural tenecin 3 . We also devised a simple and easy procedure to isolate the recombinant protein from the bacterial cell extracts . The recombinant tenecin 3 showed an antifungal activity against C . albicans as the natural tenecin 3 did . Therefore large quantities of tenecin 3 can be easily obtained by the expression and purification system of tenecin 3 described in this report.

Anesth Analg, 1999 Jan, 88(1), 209 - 12
The growth of microorganisms in propofol and mixtures of propofol and lidocaine; Wachowski I et al.; Propofol emulsion supports bacterial growth . Extrinsic contamination of propofol has been implicated as an etiological event in postsurgical infections . When added to propofol, local anesthetics (e.g., lidocaine) alleviate the pain associated with injecting it . Because local anesthetics have antimicrobial activity, we determined whether lidocaine would inhibit microbial growth by comparing the growth of four microorganisms in propofol and in mixtures of propofol and lidocaine . Known quanta of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans were inoculated into solutions of 1% propofol, 0.2% lidocaine in propofol, 0.5% lidocaine in propofol, 0.5% lidocaine in isotonic sodium chloride solution, and 0.9% isotonic sodium chloride solution . All microorganisms were taken from stock cultures and incubated for 24 h . Growth of microorganisms in each solution was compared by counting the number of colony-forming units grown from a subculture of the solution at 0, 3, 6, 12 and 24 h . Propofol supported the growth of E . coli and C . albicans . Propofol maintained static levels of S . aureus and was bactericidal toward P . aeruginosa . The addition of 0.2% and 0.5% lidocaine to propofol failed to prevent the growth of the studied microorganisms . The effect of 0.5% lidocaine in isotonic sodium chloride solution did not differ from the effects of isotonic sodium chloride solution alone . We conclude that lidocaine, when added to propofol in clinically acceptable concentrations, does not exhibit antimicrobial properties . IMPLICATIONS: Local anesthetics such as lidocaine have antimicrobial activity . Propofol supports the growth of bacteria responsible for infection . Bacteria were added to propofol and propofol mixed with lidocaine . The addition of lidocaine to propofol in clinically relevant concentrations did not prevent the growth of bacteria . The addition of lidocaine to propofol cannot prevent infection from contaminated propofol.

J Clin Microbiol, 1999 Feb, 37(2), 321 - 6
Identification of Candida dubliniensis in a prospective study of patients in the United States; Jabra-Rizk MA et al.; Although Candida albicans remains the fungal species most frequently isolated as an opportunistic oral pathogen, other yeast species are often identified in human immunodeficiency virus (HIV)-seropositive patients . Candida dubliniensis phenotypically resembles C . albicans in many respects, yet it can be identified and differentiated as a unique Candida species by its phenotypic and genetic profiles . The purpose of the present study was to prospectively test for the presence of C . dubliniensis among clinical isolates and to determine the clinical and demographic characteristics of patients harboring C . dubliniensis . Over a 90-day period, isolates from 724 patients that were presumptively identified as C . albicans were screened for C . dubliniensis by use of tests for germ tube and chlamydospore production, by detection of an inability to grow at 45 degrees C, by colony color on CHROMagar Candida medium, and by the results of a sugar assimilation test with the API 20C AUX yeast identification system . Among 699 isolates retrieved from those specimens evaluated, 5 from 25 HIV-seropositive patients and 1 isolate from a patient whose HIV status was unknown were shown to be consistent by phenotyping and by electrophoretic karyotyping with the European reference strain of C . dubliniensis . One of the C . dubliniensis isolates had dose-dependent susceptibility to fluconazole (MIC, 16 microg/ml) . These results confirm the presence of this interesting species in the United States and support the need for further investigations into the prevalence and pathogenesis of C . dubliniensis.

APMIS, 1998 Nov, 106(11), 1049 - 55
Reduced pathogenicity of a Candida albicans MAP kinase phosphatase (CPP1) mutant in the murine mastitis model; Guhad FA et al.; Candida albicans strains with a deletion of the mitogen-activated protein kinase tyrosine phosphatase gene (CPP1) are derepressed in the yeast-to-hyphal transition on solid surfaces in vitro at ambient temperatures and this gene is therefore required for repression of the yeast-to-hyphal switch . The pathology caused by a CPP1 null mutant strain was compared with that of the null mutant into which the wild-type CPP1 gene was introduced by homologous recombination and with the wild-type parent strain in a murine mycotic mastitis model . The mammary glands of lactating mice (at day 5 postpartum) were infected for 2, 4 and 6 days with 1 x 10(5), 1 x 10(6) and 1 x 10(7) cell-forming units before euthanasia . Infected and non-infected control glands were evaluated histopathologically . The null mutant strains showed less severe pathology than the two control strains . The Cpplp tyrosine phosphatase may thus be considered a virulence determinant during localized infection in C . albicans.

J Clin Microbiol, 1999 Feb, 37(2), 417 - 21
Molecular and phenotypic characterization of genotypic Candida albicans subgroups and comparison with Candida dubliniensis and Candida stellatoidea; McCullough MJ et al.; There have been increased reports of the isolation of unusual genotypic groups of Candida albicans (groups C and D) based on a well-defined genotypic method; this method uses cellular DNA digested with the EcoRI enzyme and the restriction fragment length polymorphisms (RFLPs) generated by agarose gel electrophoresis . The aim of the present study was to use additional molecular tools to characterize these unusual strains and to compare them with authentic strains of C . dubliniensis, a recently delineated species, and type I C . stellatoidea . The RFLPs of PCR products generated from the intergenic transcribed spacer (ITS) region did not differentiate among C . albicans genotypes A, B, and C and type I C . stellatoidea . However, this method did differentiate the C . albicans genotype D strains, which were identical to C . dubliniensis . The RFLPs generated by HaeIII digestion of the PCR products of the V3 region of the 25S rRNA gene (rDNA) could differentiate the same groups as RFLP analysis of the PCR amplicon of the ITS region . C . albicans genotype B isolates have been shown to have a transposable intron in the 25S rDNA, whereas genotype A isolates do not; C . dubliniensis strains also have an intron that is larger than that in genotype B C . albicans strains but that is in the same location . PCR designed to span this region resulted in a single product for C . albicans genotype A (450 bp), B (840 bp), type 1 C . stellatoidea (840 bp), and C . dubliniensis (1,080 bp), whereas the C . albicans genotype C isolates had two major products (450 and 840 bp) . All C . albicans genotype D isolates gave a PCR product identical to that given by C . dubliniensis . These results indicate that those strains previously designated C . albicans genotype D are in fact C . dubliniensis, that no differences were found between type 1 C . stellatoidea and C . albicans genotype B strains, and that the C . albicans genotype C strains appear to have the transposable intron incompletely inserted throughout the ribosomal repeats in their genomes . The results of the antifungal susceptibility testing of 105 of these strains showed that, for fluconazole, strains of C . dubliniensis were significantly more susceptible than strains of each of the C . albicans genotypes (genotypes A, B, and C) . The flucytosine susceptibility results indicated that strains of C . albicans genotype A were significantly less susceptible than either C . albicans genotype B or C . albicans genotype C strains . These results indicate that there is a correlation between the Candida groups and antifungal susceptibility.

Microbiol Immunol, 1998, 42(11), 789 - 93
Anti-Candida activity of calprotectin in combination with neutrophils or lactoferrin; Okutomi T et al.; The effect of an anti-microbial protein, calprotectin, in combination with neutrophils on the growth of Candida albicans was investigated . The growth inhibition of C . albicans by murine neutrophils was augmented by the addition of a low concentration of calprotectin prepared from rat peritoneal exudate cells . The concentrations of calprotectin causing 50% inhibition of growth of C . albicans in the absence or presence of neutrophils at an effector-to-target (E/T) ratio of 30 and 60 were estimated to be 0.45, 0.34 and 0.28 U/ml, respectively . The anti-Candida activity of calprotectin was completely inhibited by 2 microM of zinc ion, while it only partially lowered the activity of the combination of calprotectin and neutrophils . Lactoferrin, which is an anti-microbial protein released from neutrophils, strongly inhibited the growth of C . albicans in combination with calprotectin . These results suggest that calprotectin and lactoferrin released from neutrophils may cooperate to inhibit the growth of C . albicans at a local lesion of the infection where there is an accumulation of neutrophils.

Diagn Microbiol Infect Dis, 1998 Nov, 32(3), 205 - 10
In vitro interaction of fluconazole and amphotericin B administered sequentially against Candida albicans: effect of concentration and exposure time; Ernst EJ et al.; This study investigated the potential antagonism of fluconazole on amphotericin B activity against Candida albicans when administered sequentially in vitro . Yeast cells were exposed to fluconazole for time periods ranging from 0 to 24 h before the addition of amphotericin B . The combination showed fungicidal (> or = 3 log 10 reduction in CFU/mL) activity . After 4 h of exposure to fluconazole, amphotericin B activity was partially inhibited at the lower concentration tested (0.25 x MIC) . Amphotericin B activity was dramatically decreased by previous exposure to fluconazole for greater than or equal to 8 h at both the high and low concentrations tested . The activity of amphotericin B against yeast exposed to fluconazole for at least 8 h was indistinguishable from fluconazole alone and was fungistatic (< or = 2 log 10 reduction in CFU/mL) . This inhibition of amphotericin B activity persisted for a very short period (< 6 h) after removal of fluconazole from the culture medium, indicating the need for continued exposure to fluconazole for lasting inhibition of amphotericin B activity.

Cell Signal, 1998 Nov, 10(10), 713 - 9
N-acetyl-D-glucosamine induces germination in Candida albicans through a mechanism sensitive to inhibitors of cAMP-dependent protein kinase; Castilla R et al.; The present study examines the involvement of cAMP-dependent protein kinase (PKA) in the dimorphic transition of Candida albicans by assessing the in vivo effect of two permeable PKA inhibitors on N-acetyl-D-glucosamine (GlcNAc)- and serum-induced differentiation . The permeable myristoylated derivative of the heat-stable PKA inhibitor (MyrPKI), which inhibited C . albicans PKA in vitro, caused a concentration-dependent inhibition of germ-tube formation in cultures induced to germinate by GlcNAc; germination halted irrespective of the time of addition of the inhibitor . MyrPKI also blocked dibutyryl-cAMP (dbcAMP)- and glucagon-stimulated germination but did not affect serum-induced germination . H-89, another highly specific PKA inhibitor, displayed the same effect on germination . Neither MyrPKI nor H-89 had any effect on budding of yeast cells . In conclusion, our results indicate that cAMP-mediated activation of PKA plays a pivotal role in the biochemical mechanism underlying morphogenesis.

J Med Microbiol, 1998 Feb, 47(2), 99 - 102
Evaluation of phospholipase activity of Candida albicans and its correlation with pathogenicity in mice; Kothavade RJ et al.; Sixty-one isolates of Candida albicans were tested for their phospholipase activity and this parameter was correlated with pathogenicity in albino mice . Of the isolates tested, 57 (93%) showed appreciable phospholipase activity and of these, 55 (90%) were pathogenic to mice . A significant correlation was found between phospholipase activity and involvement of kidneys in animal pathogenicity studies . The isolates of C . albicans, that exhibited higher phospholipase activity were found to be pathogenic for mice.

Biochim Biophys Acta, 1999 Jan 6, 1426(2), 297 - 307
Protein O-mannosylation; Strahl-Bolsinger S et al.; Protein O-mannosylation, originally observed in fungi, starts at the endoplasmic reticulum with the transfer of mannose from dolichyl activated mannose to seryl or threonyl residues of secretory proteins . This reaction is catalyzed by a family of protein O-mannosyltransferases (PMTs), which were first characterized in Saccharomyces cerevisiae . The identification of this evolutionarily conserved PMT gene family has led to the finding that protein O-mannosylation plays an essential role in a number of physiologically important processes . Focusing on the PMT gene family, we discuss here the main aspects of the biogenesis of O-linked carbohydrate chains in S . cerevisiae, Candida albicans, and other fungi . We summarize recent work utilizing pmt mutants that demonstrates the impact of protein O-mannosylation on protein secretion, on maintenance of cell wall integrity, and on budding . Further, the occurrence of PMT orthologs in higher eukaryotes such as Arabidopsis, Drosophila and mammals is reported and discussed.

J Surg Res, 1998 Dec, 80(2), 205 - 9
Obstructive jaundice alters Kupffer cell function independent of bacterial translocation; Sheen-Chen SM et al.; BACKGROUND . There is a high incidence of perioperative morbidity and mortality in patients with obstructive jaundice . The absence of bile in the gastrointestinal tract promotes bacterial overgrowth and the increased translocation of bacteria and endotoxin to the liver which has been postulated to inhibit Kupffer cell function in these patients . But, biliary tract obstruction can directly damage liver cells and thus alter their function . Thus, we hypothesized that obstructive jaundice alone alters Kupffer cell function independent of the effects of bacterial translocation . This study was designed to evaluate the contribution of bacterial translocation to the altered Kupffer cell function observed in patients with obstructive jaundice . METHODS . Sprague-Dawley rats were randomized to three groups of six animals each . Group 1 underwent common bile duct ligation with intestinal bile salt replacement (sodium taurocholate 100 mg/kg/day) via gastrostomy and an implantable osmotic pump (CBDL + bile salts), Group 2 underwent common bile duct ligation with normal saline replacement (CBDL + saline), and Group 3 underwent a sham operation (sham control) . After 7 days, tissue and blood were collected for bacterial translocation and biochemical analyses . Examination of cultured Kupffer cell function included measuring the phagocytosis of heat-killed Candida albicans and endotoxin (LPS)-induced TNFalpha and nitric oxide production . RESULTS . While bacterial translocation and cecal bacterial counts were significantly increased in the CBDL + saline group, these parameters were both reduced to control levels following intestinal bile salt replacement (CBDL + bile salts) . Altered Kupffer cell function, as measured by the increased phagocytosis of C . albicans and LPS-induced NO production, and decreased LPS-induced TNFalpha production were observed in all animals with obstructive jaundice regardless of bile salt replacement . CONCLUSION . Kupffer cell function appears to be differentially affected by obstructive jaundice and these altered functions can occur independent of bacterial translocation .

Arch Oral Biol, 1998 Dec, 43(12), 999 - 1007
Adhesion of oral Candida albicans isolates to denture acrylic following limited exposure to antifungal agents; Ellepola AN et al.; Candidal adherence to denture acrylic surfaces is implicated as the first step in the pathogenesis of Candida-associated denture stomatitis, the most prevalent form of oral candidosis in the West . This condition is treated by topically administered antifungal agents, mainly belonging to the polyenes and azoles . As the intraoral concentrations of antifungals fluctuate considerably due to the dynamics of the oral environment, the effect of short exposure to sublethal concentrations of antifungals on the adhesion of Candida albicans to denture acrylic surfaces was investigated . Seven oral C . albicans isolates were exposed to four-eight times minimum inhibitory concentrations (MIC) of five antifungal drugs, nystatin, amphotericin B, 5-fluorocytosine, ketoconazole and fluconazole, for 1 h . After removing the drug (by repeated washing) the adhesion of these isolates to acrylic strips was assessed by an in vitro adhesion assay . Exposure to antifungal agents significantly reduced the adherence of all seven C . albicans isolates to denture acrylic . The mean percentage reductions of adhesion after limited exposure to nystatin, amphotericin B, 5-fluorocytosine, ketoconazole and fluconazole were 86.48, 90.85, 66.72, 65.88 and 47.42%, respectively . These findings indicate that subtherapeutic doses of antifungals may modulate oral candidal colonization . Further, these results may have an important bearing on dosage regimens currently employed in treating oral candidosis.

Int J Immunopharmacol, 1998 Dec, 20(12), 751 - 63
Effect of d-fenfluramine on the lymphocyte response of HIV+ humans; Mathews HL et al.; The objective of this study was to analyse the effect of d-dexfenfluramine (d-FEN) on the human lymphocyte response, in vitro . Experiments were designed to determine whether d-FEN augments specific human immune parameters associated with protection from opportunistic microbial pathogens and particularly focuses on d-FEN as a means by which to augment the function of CD8+ and CD4+ lymphocytes . Lymphocytes were examined for three reasons: (1) for their ability to inhibit the growth of Candida albicans; (2) for their ability to proliferate in response to a mitogen; and, (3) their cytokine profile (vis., production of IL-2, IFN-gamma and TNF-alpha) . Peripheral blood mononuclear cells (PBMC) were obtained from 20 HIV+ patients . The patients were diagnosed as HIV+ within the past 0.5-9 years . d-FEN was found to augment the capacity of CD8+ lymphocytes to inhibit the growth of the opportunistic microbial pathogen, C . albicans . d-FEN enhanced the capacity of CD4+ lymphocytes to proliferate in response to the mitogen, Concanavalin A, and to increase the amount of IL-2 produced by CD4+ and CD8+ lymphocytes from AIDS patients . d-FEN increased the number of CD4+ and CD8+ lymphocytes that produced IFN-gamma from either non-AIDS or AIDS patients and increased the number of AIDS patient's CD8+ lymphocytes that produce TNF-alpha . These in vitro data suggest that d-FEN may be effective in enhancing immune function in immunocompromised individuals.

Wien Klin Wochenschr, 1998 Nov 13, 110(21), 740 - 50
{New aspects in treatment of systemic mycoses}; Presterl E et al.; The incidence of systemic fungal infection has been increasing during the last two decades . Candida and Aspergillus spp . are the classical opportunistic pathogens . Rare fungi, such as Mucor, Rhizopus, Fusarium, Trichosporon, Paecilomyces, Alternaria, Cladosporium and Pseudoallescheria, are emerging as cause of systemic fungal infection in the immunocompromised host . For more than 40 years Amphotericin B has been the gold standard of antifungal treatment because of its broad spectrum comprising yeasts, dimorphic fungi and moulds . Its nephrotoxicity has led to the development of lipid-associated preparations of amphotericin B: liposomal amphotericin B, amphotericin B colloidal dispersion and amphotericin B lipid complex . These preparations are less nephrotoxic, but higher doses than those of conventional amphotericin B are needed to achieve the same effect . The triazole fluconazole is the treatment of choice in infections caused by Candida albicans . New antifungal compounds are voriconazole and the candins, the pradimicin/benanomycin family, nikkomycin Z and a liposomal preparation of nystatin.

Antimicrob Agents Chemother, 1999 Jan, 43(1), 100 - 5
Acetate-mediated growth inhibition in sterol 14alpha-demethylation-deficient cells of Candida albicans; Shimokawa O et al.; Candida albicans is a fungus thought to be viable in the presence of a deficiency in sterol 14alpha-demethylation . We showed in a strain of this species that the deficiency, caused either by a mutation or by an azole antifungal agent, made the cells susceptible to growth inhibition by acetate included in the culture medium . Studies with a mutant demonstrated that the inhibition was complete at a sodium acetate concentration of 0.24 M (20 g/liter) and was evident even at a pH of 8, the latter result indicating the involvement of acetate ions rather than the undissociated form of acetic acid . In fluconazole-treated cells, sterol profiles determined by thin-layer chromatography revealed that the minimum sterol 14alpha-demethylation-inhibitory concentrations (MDICs) of the drug, thought to be the most important parameter for clinical purposes, were practically identical in the media with and without 0.24 M acetate and were equivalent to the MIC in the acetate-supplemented medium . The acetate-mediated growth inhibition of azole-treated cells was confirmed with two additional strains of C . albicans and four different agents, suggesting the possibility of generalization . From these results, it was surmised that the acetate-containing medium may find use in azole susceptibility testing, for which there is currently no method capable of measuring MDICs directly for those fungi whose viability is not lost as a result of sterol 14alpha-demethylation deficiency . Additionally, the acetate-supplemented agar medium was found to be useful in detecting reversions from sterol 14alpha-demethylation deficiency to proficiency.

J Nat Prod, 1998 Dec, 61(12), 1539 - 42
New cyclic peroxides from the Philippine sponge Plakinastrella sp; Qureshi A et al.; Three new cyclic peroxides 5-7 and a new carboxylic acid ester 8 were isolated as minor metabolites from the hexane extract of a Plakinastrella species from the Philippines . The structures of compounds 5-8 were elucidated by interpretation of spectral data and by chemical interconversion, and the absolute stereochemistry of peroxide 6 was determined by application of Mosher's method to a derivative . Although the major compounds in the sponge showed activity against Candida albicans prior to decomposition, the minor metabolites 5-8 are essentially inactive.

Am J Hosp Palliat Care, 1998 Nov-Dec, 15(6), 315 - 9
Fluconazole sensitivities of Candida species isolated from the mouths of terminally ill cancer patients; Ball K et al.; Oral candidosis is common in advanced cancer and is often treated with the systemic triazole antifungal drug fluconazole . This study examined the species of yeast present in the mouths of 30 patients with advanced cancer and determined their sensitivity to fluconazole . Thirty-five yeast isolates were collected from a total of 25 (83 percent) of the patients sampled . The two most common species were Candida albicans (15 isolates) and C . glabrata (11 isolates)--with smaller numbers of C . tropicalis, C . parapsilosis, C . guilliermondii, C . inconspicua, and Saccharomyces cerevisiae . The minimal inhibitory concentrations (MIC) of fluconazole for the strains of C . albicans were generally low (median 0.19 microgram/ml) but were considerably higher for C . glabrata (median 2 micrograms/ml) . The remaining species demonstrated MICs similar to those for C . albicans, with the exceptions of C . inconspicua and Saccharomyces cerevisiae, which were relatively insensitive . In conclusion, non-albicans yeasts are common in the mouths of patients with advanced cancer and these may have reduced sensitivity to fluconazole . Mycological diagnosis is a valuable aid to management.

Immunol Lett, 1998 Nov, 64(1), 23 - 9
Identification of a soluble interleukin-8 inhibitor in the supernatant of polymorphonuclear leukocytes; Kemeny L et al.; Interleukin-8 (IL-8) plays a crucial role in the pathogenesis of inflammatory and hyperproliferative diseases in various organs . The purpose of the present investigation was to establish whether there is any naturally occurring inhibitor of IL-8 . Here we demonstrate that an IL-8 inhibitor (IL-8INH) is present in the supernatant of polymorphonuclear (PMN) leukocytes . The release of IL-8INH could be increased by stimulating the PMN leukocytes by concanavalin A . IL-81NH blocks the IL-8-induced chemotaxis and Candida albicans killing activity of PMN leukocytes and epidermal cells in vitro, and IL-8-induced neutrophil infiltration in the mouse ear in vivo . The mechanism of action of IL-8INH involves blocking of 125I-IL-8 binding to the IL-8 receptor . Binding of 125I-IL-8 to neutrophils could not be displaced by the IL-8INH, however, preincubation of 125I-IL-8 with IL-8INH increased binding inhibition, suggesting an interaction between IL-8 and the inhibitor . Crosslinking of 125I-IL-8 to IL-8INH shows that IL-8INH binds specifically to 125I-IL-8, and the IL-8INH protein has an apparent molecular weight of 52 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The partial purification of the IL-8INH on DEAE-Sephadex anion-exchange chromatography column also suggests a 50-60-kDa inhibitor protein which blocks IL-8-induced effects on neutrophils by binding to IL-8.

J Bacteriol, 1999 Jan, 181(1), 231 - 40
Isolation of a putative Candida albicans transcriptional regulator involved in pleiotropic drug resistance by functional complementation of a pdr1 pdr3 mutation in Saccharomyces cerevisiae; Talibi D et al.; Three Candida albicans genes, designated FCR (for fluconazole resistance), have been isolated by their ability to complement the fluconazole (FCZ) hypersensitivity of a Saccharomyces cerevisiae mutant lacking the transcription factors Pdr1p and Pdr3p . Overexpression of any of the three FCR genes in the pdr1 pdr3 mutant resulted in increased resistance of the cells to FCZ and cycloheximide and in increased expression of PDR5, a gene coding for a drug efflux transporter of the ATP-binding cassette superfamily and whose transcription is under the control of Pdr1p and Pdr3p . Deletion of PDR5 in the pdr1 pdr3 strain completely abrogated the ability of the three FCR genes to confer FCZ resistance, demonstrating that PDR5 is required for FCR-mediated FCZ resistance in S . cerevisiae . The FCR1 gene encodes a putative 517-amino-acid protein with an N-terminal Zn2C6-type zinc finger motif homologous to that found in fungal zinc cluster proteins, including S . cerevisiae Pdr1p and Pdr3p . We have constructed a C . albicans CAI4-derived mutant strain carrying a homozygous deletion of the FCR1 gene and analyzed its ability to grow in the presence of FCZ . We found that the fcr1Delta/fcr1Delta mutant displays hyperresistance to FCZ and other antifungal drugs compared to the parental CAI4 strain . This hyperresistance could be reversed to wild-type levels by reintroduction of a plasmid-borne copy of FCR1 into the fcr1Delta/fcr1Delta mutant . Taken together, our results indicate that the FCR1 gene behaves as a negative regulator of drug resistance in C . albicans and constitute the first evidence that FCZ resistance can result from the inactivation of a regulatory factor such as Fcr1p.

Wei Sheng Wu Xue Bao, 1997 Jun, 37(3), 212 - 6
{Screening of Candida albicans fluconazole--resistant mutation strains}; Wang W et al.; We adopted the method of miminum inhibitary concentration (MIC) assay according to the method recommened by National Committee for Clinical Laboratory Standard (NCCLS), and selected 2 fluconazole--susceptible strains from over 50 strains of Candida albicans which were preserved in our center (these strains were all isolated from clinical patients) . The number of the strains are: BMU8945 (MIC value is 0.125 microgram/ml) and BMU8977 (MIC value is 0.25 microgram/ml) . In addition, we also picked one strain of ATCC14053 (MIC value is 0.5 microgram/ml) as our research starting strains . After the induction of UV and DES, we obtained nearly 1000 colonies of fluconazole--resistant strains (MIC > 30 micrograms/ml) . After subculture, we found that the characterization of drug--resistance of these strains could be inheredited stably.

Mycoses, 1998 Sep-Oct, 41(7-8), 327 - 34
Fungal colonization of the stomach and its clinical relevance; Zwolinska-Wcislo M et al.; The aim of this study was to estimate the frequency of fungal colonization of the stomach of patients suffering from gastric ulcer (GU) and chronic gastritis (CG) and the influence of fungal colonization of the stomach on the process of ulcer healing . We investigated 293 patients aged 20-80 years . Before and after 4 weeks of sucralfate treatment they underwent endoscopy of the stomach, histological examination of biopsies taken from the ulcer margin or inflamed gastric mucosa and mycological examinations of the gastric juice, surface brushing and biopsies . The studies revealed a high concentration of fungi in 54.2% patients with GU and 10.3% with CG . Candida albicans was the most frequently isolated organism . Fungal colonization of the stomach impairs the process of gastric ulcer healing . Control examination after 4 weeks of sucralfate therapy showed the ratio of GU healing in 62% of patients with a high concentration of fungi in comparison with 78% of patients not colonized with fungi (P < 0.05) . A significantly longer duration of ulcer symptoms in the group of patients with a high concentration of fungi in the stomach was also observed . There was no correlation between the level of fungal antibodies, of Candida antigen in the serum and the concentration of fungi in the stomach.

Mycoses, 1998 Sep-Oct, 41(7-8), 321 - 5
HIV protease inhibitors influence the prevalence of oral candidosis in HIV-infected patients: a 2-year study; Hoegl L et al.; The introduction of HIV protease inhibitors was accompanied by reduction in HIV-associated opportunistic infections . Therefore, we performed a retrospective study of HIV-infected patients to evaluate the effects of therapy with an HIV protease inhibitor (PI) on oral candidosis . This was of special interest, because an important virulence factor of Candida albicans is the secreted aspartic protease (SAP), which is assigned to the same class of aspartic proteases as HIV protease . Sixty-two patients were examined five times over a period of 2 years . There was a hint at a difference in the frequencies of C . albicans carrier state and manifest oral candidosis in favour of treatment with a PI . In addition, loss of Candida colonization and manifest oral candidosis was observed only in patients with elevation of CD4 cells upon PI . This might explain the effect, which also might go back to a direct inhibition of yeast SAP.

Dis Colon Rectum, 1998 Dec, 41(12), 1581 - 4
Fungal sacral osteomyelitis as the initial presentation of Crohn's disease of the small bowel: report of a case; Armstrong N et al.; We report a unique case of Candida albicans sacral osteomyelitis in a 48 year-old female with previously undiagnosed Crohn's disease . The patient was ill for one year with fatigue, weakness, and a 60-lb weight loss . At the time of presentation, she developed chills, fever, right lower quadrant abdominal pain, and right knee pain . Physical examination was significant for a palpable right lower quadrant abdominal mass . A computed tomographic scan of the abdomen and pelvis identified a large right-sided retroperitoneal mass, severe right hydronephrosis, and air within the right sacrum . Findings at laparotomy included small-bowel changes consistent with Crohn's disease, a multiloculated retroperitoneal abscess, and evidence of sacral osteomyelitis . A right hemicolectomy with sacral debridement and placement of presacral drains was performed . Bone cultures from the sacrum demonstrated a predominance of C . albicans, in addition to coliforms and enterococcus . The patient was placed on amphotericin B and intravenous antibiotics . Because serial computed tomographic scans of her pelvis demonstrated progression of her pelvic osteomyelitis to include the sacrum, right ilium, right acetabulum, and right femoral head, a repeat debridement with resection of the right femoral head was performed . After 12 months of follow-up, she was doing well without medications and had no constitutional symptoms or radiographic evidence of disease progression . This report illustrates a unique case of Crohn's disease presenting as sacral osteomyelitis secondary to small-bowel fistulization . Aggressive multidisciplinary surgical and medical management were the key to the successful management of this difficult case.

Arch Dermatol Res, 1998 Nov, 290(11), 603 - 9
The importance of CD54 and CD86 costimulation in T cells stimulated with Candida albicans and Dermatophagoides farinae antigens in patients with atopic dermatitis; Kawamura MS et al.; A number of studies have demonstrated an increased frequency of allergen-specific T cells producing increased amounts of interleukin-4 (IL-4) and IL-5, but little interferon-y in both the peripheral blood and skin lesions of patients with atopic dermatitis (AD) . In this study, to further clarify the characteristics of T cells obtained from AD patients, we examined the dependency of the antigen-specific proliferation of peripheral blood mononuclear cells (PBMC) from AD patients on costimulatory molecules . The antigens used were Candida albicans and Dermatophagoides farinae, for which AD patients show increased levels of IgE antibodies . PBMC from control healthy donors stimulated with these antigens incorporated {3H}-thymidine much more than PBMC from AD patients . The addition of anti-CD54, -CD40, -CD80 and -CD86 monoclonal antibodies to the cultures showed that the PBMC required only CD54 and CD86 for stimulation with C . albicans, but required CD54, CD80 and CD86 for stimulation with D . farinae . Among these monoclonal antibodies, the anti-CD54 antibody suppressed the proliferative responses of most PBMC, most effectively followed by the anti-CD86 antibody . However, there were no significant differences in the requirement for costimulatory molecules of PBMC proliferation stimulated with C . albicans or D . farinae between AD patients and healthy donors . Since many studies have suggested that T-helper type 1 and T-helper type 2 immune responses are different in their dependency on CD80 or CD86 costimulation, our present results suggest that the allergen-specific T cells of AD patients are not completely shifted to a T-helper type 2 subset.

Rev Soc Bras Med Trop, 1998 Nov-Dec, 31(6), 523 - 7
{Killer toxin and enzyme production by Candida albicans isolated from buccal mucosa in patients with cancer}; de Oliveira EE et al.; Opportunistic infections of the oral cavity are primarily caused by Candida and frequently occur in patients with cancer who are undergoing chemotherapy and antibiotic treatment . Of the specimens received from the oral mucosa of 44 patients with cancer, 25 (56.8%) yielded Candida on culture in Sabouraud agar . Twenty four of these isolates were identified as C . albicans (96%) and 1 as C . krusei (4%) . The phenotypic characteristics of these isolates showed that all of them were strongly proteolytic, had a high ability to produce phospholipase, and presented the byotypes characterized as 811 (95.8%) and 511 (4.2%) in terms of susceptibility to killer toxins.

Anticancer Res, 1998 Sep-Oct, 18(5B), 3629 - 38
Anti-infective catheters: novel strategies to prevent nosocomial infections in oncology; Schierholz JM et al.; Intravenous access contributes significantly to the therapeutical success and to the comfort of oncologic patients . The highest risk for bloodstream infections, however, is vascular catheter-mediated . In oncology high mortality is associated with Pseudomonas aeruginosa, Candida albicans and Staphylococcus aureus sepsis . Besides established hygienic measures, the coupling or incorporation of antimicrobial substances to or into catheter materials may be a suitable way to prevent the development of catheter-associated infections . Here we present a risk- benefit evaluation of different models of antimicrobial catheter coated with silver, antiseptics or antibiotics . The controversial reports on clinical efficacy and the potential of adverse reactions due to silver and antiseptic coated catheters are discussed . The microbiological, pharmaceutical and physicochemical backgrounds of different types of coating are discussed in detail . Incorporation of antimicrobial agents into long-term silicon catheters providing a slow release of those substances through the external and internal surfaces of catheters may be the most effective technological innovation for reducing biomaterial-mediated nosocomial infections.

Pediatr Infect Dis J, 1998 Nov, 17(11), 1012 - 5
Candida fungemia in neonates treated with fluconazole: report of forty cases, including eight with meningitis; Huttova M et al.; PURPOSE OF THE STUDY: To assess efficacy and safety of fluconazole in neonates with Candida fungemia . STUDY DESIGN: Multicenter prospective protocol of all fungemias appearing between January 1, 1993, and December 31, 1997, in four major university hospitals . RESULTS: Forty neonates, 28 of them with very low birth weight (<1500 g; 30.5 median gestation week), with documented Candida albicans fungemia were treated with intravenous fluconazole in a daily dosage of 6 mg/kg once daily for 6 to 48 days . Thirty-four received fluconazole as monotherapy and 6 received it in combination with amphotericin B . Thirty-two (80%) were cured; 4 of them relapsed despite at least 14 days of therapy, but they were ultimately cured without sequelae . Eight other neonates died, 4 because of fungal infection and 4 because of prematurity or hemorrhage or lung failure, with fungemia (20% overall and 10% attributable mortality) . Two neonates had elevated liver enzymes during fluconazole therapy and 2 others had elevated serum creatinine during fluconazole monotherapy . In none of them did these abnormalities necessitate discontinuation of antifungal therapy . In 8 neonates fungal meningitis developed as a complication of fungemia . All but 3 fungemias were C . albicans; 3 were Candida parapsilosis . CONCLUSIONS: Fluconazole was safe and effective antifungal therapy even in complicated or Candida fungemia in neonates and in infants with very low birth weight.

J Bacteriol, 1998 Dec, 180(24), 6607 - 16
A MADS box protein consensus binding site is necessary and sufficient for activation of the opaque-phase-specific gene OP4 of Candida albicans; Lockhart SR et al.; The majority of strains of Candida albicans can switch frequently and reversibly between two or more general phenotypes, a process now considered a putative virulence factor in this species . Candida albicans WO-1 switches frequently and reversibly between a white and an opaque phase, and this phenotypic transition is accompanied by the differential expression of white-phase-specific and opaque-phase-specific genes . In the opaque phase, cells differentially express the gene OP4, which encodes a putative protein 402 amino acids in length that contains a highly hydrophobic amino-terminal sequence and a carboxy-terminal sequence with a pI of 10.73 . A series of deletion constructs fused to the Renilla reniformis luciferase was used to functionally characterize the OP4 promoter in order to investigate how this gene is differentially expressed in the white-opaque transition . An extremely strong 17-bp transcription activation sequence was identified between -422 and -404 bp . This sequence contained a MADS box consensus binding site, most closely related to the Mcm1 binding site of Saccharomyces cerevisiae . A number of point mutations generated in the MADS box consensus binding site as well as a complete deletion of the consensus site further demonstrated that it was essential for the activation of OP4 transcription in the opaque phase . Gel mobility shift assays with the 17-bp activation sequence identified three specific complexes which formed with both white- and opaque-phase cell extracts . Competition with a putative MADS box consensus binding site from the promoter of the coordinately regulated opaque-phase-specific gene PEP1 (SAP1) and the human MADS box consensus binding site for serum response factor demonstrated that one of the three complexes formed was specific to the OP4 sequence.

Res Immunol, 1998 Sep-Oct, 149(7-8), 643 - 6
Monocyte-derived dendritic cells: development of a cellular processor for clinical applications; Goxe B et al.; Since dendritic cells (DCs) are the most professional antigen-presenting cells, (Schuler et al., 1997), increasing interest in their use in clinical approaches has been observed . (Nestle et al., 1998; Murphy G . et al., 1996) . We have developed an ex vivo standardized process for the generation of dendritic-like cells (MAC-DCs) from human blood circulating monocytes . Human monocytes can differentiate into very different functional cells according to the conditions of culture, media and cytokines used . In the present study, we demonstrate that both pure monocytes and mononuclear cells differentiate into DCs when they are grown in defined medium AIM-V in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) plus IL13 and in approved biocompatible non-adherent bags . Quality and functional controls of the immature DCs obtained rely on bacterial sterility, viability, morphology and recovery . The MAC-DCs also present an immature DC phenotype with a low expression of CD14 and CD64, and high expression of MHC-I, MHC-II and CD40 . They also express B7 costimulatory molecules (CD80, CD86), CD83, and CD1a molecules . They induce strong allogenic T-cell proliferation (mixed lymphocyte reaction as well as proliferation of autologous memory T lymphocytes when incubated in the presence of recall antigens (tuberculosis, Candida albicans, and tetanus toxoid) . They also show an increase in phagocytic uptake of yeast, tumour cells and debris . The global closed system which, under reproducible good medical practice (GMP) conditions, enables the production of dendritic cells of clinical quality, has been optimized ("Vac Cell Processor") . It contains all bags, connections, media, reagents, washing solutions, control antibodies, standard operating procedures, data management, traceability and help in the form of dedicated software.

FEMS Microbiol Lett, 1998 Dec 1, 169(1), 131 - 8
Nature of Candida albicans-derived carbohydrate antigen recognized by a monoclonal antibody in patient sera and distribution over Candida species; Jacquinot PM et al.; The minimal epitope of an anti-Candida albicans mannan monoclonal antibody (MAb) EB-CA1, used to detect mannanemia in patient sera, was determined, MAb EB-CA1 exhibited reactivity with oligomannosides released from the mannan acid stable domain, converted into neoglycolipids (NGLs) and coated onto ELISA plates . Reactivity occurred with mannopentaose and higher oligomers, whereas mannotriose and mannotetraose were unreactive . MAb EB-CA1 binding to mannan acid stable mannopentaose NGL displayed a dose dependent and saturable specific reactivity curve whereas there was a complete absence of binding, even at high concentrations, with NGLs constructed from the beta-1,2-linked mannopentaose derived from the mannan acid labile fraction . MAb EB-CA1 binding to acid stable mannopentaose NGL was inhibited by the homologous oligomannoside but not by mannotriose and mannotetraose . NMR analysis showed that mannotriose and mannotetraose contained exclusively alpha-1,2-linked D-mannopyranose units and that mannopentaose was a mixture of a mannopentaose alpha-1,2-linked and an isomer in which the fifth mannose was alpha-1,6-linked to the reducing unit of manno-alpha-1,2 tetraose . Western blot analysis has shown that MAb EB-CA1 epitope was expressed on a wide range of C . albicans manno-glycoconjugate as well as on manno-glycoconjugates of other pathogenic species of the genus Candida, viz . C . tropicalis, C . glabrata, C . parapsilosis and C . krusei.

Antonie Van Leeuwenhoek, 1998 May, 73(4), 373 - 80
Biosynthesis of glycoproteins in Candida albicans: biochemical characterization of dolichol phosphate glucose synthase; Rodriguez-Bonilla J et al.; A mixed membrane fraction isolated from C . albicans yeast cells catalyzed the transfer of glucose from UDP-Glc into three classes of endogenous acceptors: glucolipid, glycoprotein and lipid-linked oligosaccharides . About 80% of the total radioactivity transferred into these products corresponded to the glucolipid which was identified as dolichol phosphate glucose by several criteria . The remainder was detected in about equal proportions in the other two fractions . Conditions that stimulated or inhibited glucolipid synthesis did not affect the extent of glycoprotein labeling . The synthesis of dolichol phosphate glucose exhibited a K(m) of 104 microM UDP-Glc and was stimulated by Mg2+ but not by Mn2+ or Ca2+ . The latter cations were, however, better stimulators of glycoprotein labeling than Mg2+ . Most nucleotides strongly inhibited the synthesis of dolichol phosphate glucose, UMP being a competitive inhibitor with a Ki of 100 microM . The dolichol phosphate glucose synthase reaction was reversed about 57% by 0.62 mM UDP but not by UMP.

Antonie Van Leeuwenhoek, 1998 May, 73(4), 289 - 97
Biosynthesis of glycoproteins in Candida albicans: solubilization and partial characterization of dolichol phosphate mannose synthase and protein mannosyl transferases; Arroyo-Flores BL et al.; Incubation of a mixed membrane fraction isolated from C . albicans yeast cells with Nonidet P-40 at a detergent/protein ratio as low of 0.025 (0.016-0.019%, w/v) yielded a soluble fraction that catalyzed the transfer of mannose from GDP-{14C} Man into dolichol phosphate mannose and from this intermediate into mannoproteins . Over 95% of the sugar in mannoproteins was O-linked as judged from its release after beta-elimination . Mannose was identified as the sole product after this treatment . Transfer activity did not depend on exogenous lipid acceptor indicating that the latter was solubilized along with the mannosyl transferases . Synthesis of mannolipid and mannoproteins occurred at optima temperatures of 20 degrees C, and 37 degrees C, respectively, and at a pH in the range of 7.5-9.5 . Mannosyl transfer into the mannolipid was stimulated by Mg2+ and inhibited by Ca2+ and Mn2+ whereas mannoprotein labeling was stimulated by Mn2+ and to a lower extent by Mg2+ . When measured as a function of substrate concentration, the synthesis of the mannolipid was a nearly linear function of GDP-Man concentration in the range of 5 to 32 microM whereas protein mannosylation exhibited hyperbolic kinetics with saturation reached at about 10 microM . The solubilized preparation was able to utilize an exogenous source of mannolipid as sugar donor for protein mannosylation . Dinucleotides and, to a higher extent trinucleotides, inhibited mannosyl transfer into the mannolipid and hence into mannoproteins.

J Ethnopharmacol, 1998 Oct, 62(3), 243 - 6
Preliminary antifungal and cytotoxicity studies of extracts of Ritchiea capparoides var . longipedicellata; Ajaiyeoba EO et al.; Crude extracts obtained from the leaves, stem bark and roots of Ritchiea capparoides var . longipedicellata were screened for in vitro antifungal activity using the agar tube dilution method . The leaf hexane, leaf methanol, stem bark methanol and root methanol extracts were tested using ten clinical strains of fungi at a concentration of 200 and 400 microg/ml, respectively . At 400 microg/ml, all four extracts inhibited the growth of six of the ten test fungi used in the study . Inhibition of the growth of Aspergillus niger by the extracts was also seen but the activity was low and the leaf hexane and root methanol extracts inhibited the growth of Drechslera rostrata . Only the leaf hexane extract was active against Curvularia lunata, while the growth of Candida albicans was not inhibited by any of the extracts . The inhibition of growth of almost all the microorganisms decreased at 200 microg, griseofulvin was included as a reference compound and methanol as the control . Preliminary cytotoxicity tests were done with the four extracts using the larvae of the brine shrimp, Artemia saline . The extracts were however found to be relatively non-toxic as each extract had an LD50 value greater than 1000 microg/ml.

J Antimicrob Chemother, 1998 Nov, 42(5), 591 - 5
In-vitro activity of essential oils, in particular Melaleuca alternifolia (tea tree) oil and tea tree oil products, against Candida spp; Hammer KA et al.; The in-vitro activity of a range of essential oils, including tea tree oil, against the yeast candida was examined . Of the 24 essential oils tested by the agar dilution method against Candida albicans ATCC 10231, three did not inhibit C . albicans at the highest concentration tested, which was 2.0% (v/v) oil . Sandalwood oil had the lowest MIC, inhibiting C . albicans at 0.06% . Melaleuca alternifolia (tea tree) oil was investigated for activity against 81 C . albicans isolates and 33 non-albicans Candida isolates . By the broth microdilution method, the minimum concentration of oil inhibiting 90% of isolates for both C . albicans and non-albicans Candida species was 0.25% (v/v) . The minimum concentration of oil killing 90% of isolates was 0.25% for C . albicans and 0.5% for non-albicans Candida species . Fifty-seven Candida isolates were tested for sensitivity to tea tree oil by the agar dilution method; the minimum concentration of oil inhibiting 90% of isolates was 0.5% . Tests on three intra-vaginal tea tree oil products showed these products to have MICs and minimum fungicidal concentrations comparable to those of non-formulated tea tree oil, indicating that the tea tree oil contained in these products has retained its anticandidal activity . These data indicate that some essential oils are active against Candida spp., suggesting that they may be useful in the topical treatment of superficial candida infections.

Fiziol Zh, 1998, 44(4), 3 - 9
{The adjuvant and specific activity of transfer factors to Candida albicans antigens}; Borysov VA et al.; Activity of transfer-factor (TF) delayed type hypersensitivity in allogenic and xenogeneic have studied . Guinea pigs was been sensibilized Soluble antigens and whole cells Candida albicans for production TF . Both TF expressed non specific and specific activity in the guinea pig and mouse leucocytes inhibition migration in vitro and delayed type hypersensitivity in mica . Activity of TF to cellular antigen was lower then TF to soluble antigen in xenogeneic systems . However TF activity was some low for whole cells in xenogeneic systems.

Microbiology, 1998 Nov, 144 ( Pt 11), 3171 - 80
A 1,3-beta-glucanosyltransferase isolated from the cell wall of Aspergillus fumigatus is a homologue of the yeast Bgl2p; Mouyna I et al.; A 1,3-beta-glucanosyltransferase which introduces intrachain 1,6-beta linkages into 1,3-beta-glucan was isolated from the cell wall of Aspergillus fumigatus . The biochemical and molecular characterization of the A . fumigatus transferase showed it was homologous to the Saccharomyces cerevisiae and Candida albicans transferase Bgl2p . A null mutant constructed in A . fumigatus by gene replacement did not show a distinct phenotype from the parental strain . The putative function of this major cell-wall-associated protein is discussed.

Microbiology, 1998 Nov, 144 ( Pt 11), 2951 - 60
A Candida albicans chaperonin subunit (CaCct8p) as a suppressor of morphogenesis and Ras phenotypes in C . albicans and Saccharomyces cerevisiae; Rademacher F et al.; Saccharomyces cerevisiae and the pathogen Candida albicans can be induced to undergo morphogenesis from a yeast to a filamentous form . A C . albicans gene (CaCCT8) was identified encoding a subunit of the Cct chaperonin complex, whose expression prevents filament formation in both fungi without interfering with growth of the yeast form . In S . cerevisiae, pseudohyphal growth induced by Ras2Val19, by overproduction of Phd1p or by expression of the C . albicans EFG1 gene, was blocked by CaCct8p and its N-terminally deleted derivative CaCct8-delta1p; in contrast, pseudohyphal induction by other components (Cph1p, Cdc42p) could not be suppressed, indicating that morphogenesis per se is not inhibited . CaCCT8 expression also interfered with other Ras2pVal19 phenotypes, including heat sensitivity, lack of glycogen accumulation and lack of sporulation . In C . albicans, overproduction of CaCct8p effectively blocked hyphal morphogenesis induced by starvation conditions and by serum . The results suggest that the activity of a component in the Ras2p signal transduction pathway is suppressed by excess chaperonin subunits . This component may be a novel folding target for the Cct complex . In agreement with this hypothesis, disruption of one of the two CaCCT8 alleles in C . albicans led to defective hyphal morphogenesis.

J Infect Dis, 1999 Jan, 179(1), 201 - 8
Evidence that members of the secretory aspartyl proteinase gene family, in particular SAP2, are virulence factors for Candida vaginitis; De Bernardis F et al.; Virulence of Candida albicans strains with targeted disruption of secretory aspartyl proteinase genes (SAP1 to SAP6) was assessed in an estrogen-dependent rat vaginitis model . Null sap1 to sap3 but not sap4 to sap6 mutants lost most of the virulence of their parental strain SC5314 . In particular, the sap2 mutant was almost avirulent in this model . Reinsertion of the SAP2 gene into this latter mutant led to the to recovery of the vaginopathic potential . The vaginal fluids of the animals infected by the wild type strain or by the sap1 or sap3 mutants expressed a pepstatin-sensitive proteinase activity in vitro . No traces of this activity were found in the vaginal fluid of rats challenged by the sap2 mutant . All strains were capable of developing true hyphae during infection . Thus, members of SAP family, in particular SAP2, play a clear pathogenic role in vaginitis and may constitute a novel target for chemoimmunotherapy of this infection.

Rev Cubana Med Trop, 1998, 50(1), 48 - 53
{Minimum inhibitory concentration of amphotericin B in yeasts of medical interest}; Fernandez Andreu CM et al.; In order to know the sensitivity of Candida and Crytococcus to amphotericin B, main drug for the treatment od systemic mycosis, it was determined the minimum inhibitory concentration (MIC) in 90 clinical isolates of Candida and Crytococcus by a micromethod for the broth dilution . According to the results, Crytococcus neoformas was more sensitive then Candida albicans (geometrical means 0.24 and 0.41 respectively) . Only one resistant strain was found (CMI = 16 micrograms/mL), corresponding to the Candida krusei species . The introduction of this technique in the Mycology Laboratory of the "Pedro Kouri" Institute of Tropical Medicine will allow to establish the sensitivity patterns and to detect the possible appearance of resistance in the main species of pathogenic fungus for men in our environment.

J Med Microbiol, 1998 Jul, 47(7), 623 - 8
An evaluation of the cost-effectiveness of using CHROMagar for yeast identification in a routine microbiology laboratory; Ainscough S et al.; CHROMagar, a chromogenic differential culture medium, is claimed to facilitate the isolation and presumptive identification of certain clinically important yeast species, e.g., Candida albicans . This study evaluated the cost-effectiveness and time advantage of using it in comparison with Sabouraud dextrose agar (SDA) . Three possible pathways, each of which included the use of one or both media, were compared in a routine laboratory . A total of 21 yeast isolates was cultured from 298 clinical samples from neutropenic and AIDS patients . An overall sensitivity of 95.2% was observed for each medium and primary isolation on CHROMagar was found to be 100% sensitive and 100% specific for C . albicans . For identification purposes, after initial culture the use of CHROMagar provided the most economical and least time-consuming method . Direct inoculation on to CHROMagar is recommended for blood cultures when yeast cells are seen on microscopy and where early appropriate therapy is imperative.

Pathology, 1998 Nov, 30(4), 405 - 18
Molecular methods for epidemiological and diagnostic studies of fungal infections; Gottfredsson M et al.; Over the past two decades there has been a remarkable increase in the incidence of invasive fungal infections . Molecular methods, such as karyotyping, restriction analysis and polymerase chain reaction (PCR), have now been applied to improve our current understanding of the epidemiology of these fungal infections . For example, investigations on nosocomial outbreaks of fungal infections have been greatly facilitated by molecular methods . In addition, the ability to diagnose and identify deep-seated mycoses may be enhanced by the use of molecular techniques . In the near future it is possible that PCR-based methods will supplement, or perhaps even replace, traditional methods for detection of Candida albicans blood stream infections, invasive aspergillosis and Pneumocystis carinii pneumonia . This review examines the progress of molecular biology into the clinical arena of fungal epidemiology, laboratory identification and diagnosis.

Minerva Stomatol, 1998 Sep, 47(9), 351 - 9
{Candida albicans: the relationship between morphotypes and in-vitro adhesivity to the oral mucosa cells of HIV-positive and AIDS patients . I}; Malighetti V et al.; BACKGROUND: The significant frequency of oropharyngeal candidiasis due to C . albicans in HIV-infected and AIDS patients and the undoubted differences in pathogenicity among strains, lead us to study whether a possible correlation exists between the phenotypic characteristics of the fungal strain and the blastospores adhesivity to the human buccal epithelial cells . METHODS: From 203 oro-pharyngeal swabs of HIV-infected patients, 133 C . albicans were identified among 159 yeast strains . Analytical strains delineation below the species level was made by the morphotyping method, assigning a morphotype code of three digits, each of which being expressive of one colonial fringe characteristic . To study the possible blastospores adhesivity differences, we have isolated 10 strains of C . albicans, chose according to the more significant colonial morphologies; they were mixed with oral epithelial cells obtained by scraping of 36 HIV-positive patients and 2 volunteer donors HIV-negative at the rate cells/blastospores of 1:20 . Suspensions were filtered, fixed and examined by optical microscope (MO) for counting the number of blastospores adhering to 100 epithelial cells and the number of cells with adhering blastospores . RESULTS: The index obtained by comparing the two qualitative analysis was higher for these isolates producing a rough or very coarse lateral fringes . CONCLUSIONS: This finding suggests that these strains may possess the highest adhesive properties, in fact the index decreases progressively to reach lower values for the strain not producing fringe.

Antimicrob Agents Chemother, 1998 Dec, 42(12), 3242 - 4
In vitro susceptibilities of Candida bloodstream isolates to the new triazole antifungal agents BMS-207147, Sch 56592, and voriconazole; Pfaller MA et al.; BMS-207147, Sch 56592, and voriconazole are three new investigational triazoles with broad-spectrum antifungal activity . The in vitro activities of these three agents were compared with those of itraconazole and fluconazole against 1,300 bloodstream isolates of Candida species obtained from over 50 different medical centers in the United States . The MICs of all of the antifungal drugs were determined by broth microdilution tests performed according to the National Committee for Clinical Laboratory Standards method using RPMI 1640 as a test medium . BMS-207147, Sch 56592, and voriconazole were all quite active against all Candida sp . isolates (MICs for 90% of the isolates tested {MIC90s}, 0.5, 1.0, and 0.5 microgram/ml, respectively) . Candida albicans was the most susceptible species (MIC90s, 0.03, 0.06, and 0.06 microgram/ml, respectively), and C . glabrata was the least susceptible (MIC90s, 4 . 0, 4.0, and 2.0 microgram/ml, respectively) . BMS-207147, Sch 56592, and voriconazole were all more active than itraconazole and fluconazole against C . albicans, C . parapsilosis, C . tropicalis, and C . krusei . There existed a clear rank order of in vitro activity of the five azoles examined in this study when they were tested versus C . glabrata: voriconazole > BMS-207147 = Sch 56592 = itraconazole > fluconazole (MIC90s, 2.0, 4.0, 4.0, 4.0, and 64 microgram/ml, respectively) . For isolates of Candida spp . with decreased susceptibility to both itraconazole and fluconazole, the MICs of BMS-207147, Sch 56592, and voriconazole were also elevated . These results suggest that BMS-207147, Sch 56592, and voriconazole all possess promising antifungal activity and that further in vitro and in vivo investigations are warranted to establish the clinical value of this improved potency.

Antimicrob Agents Chemother, 1998 Dec, 42(12), 3065 - 72
Multiple molecular mechanisms contribute to a stepwise development of fluconazole resistance in clinical Candida albicans strains; Franz R et al.; From each of two AIDS patients with oropharyngeal candidiasis, five Candida albicans isolates from recurrent episodes of infection which became gradually resistant against fluconazole during antimycotic treatment were analyzed for molecular changes responsible for drug resistance . In both patients, a single C . albicans strain was responsible for the recurrent infections, but the CARE-2 fingerprint pattern of the isolates exhibited minor genetic alterations, indicating that microevolution of the strains took place during fluconazole therapy . In the isolates from patient 1, enhanced mRNA levels of the MDR1 gene, encoding a multiple drug resistance protein from the superfamily of major facilitators, and constitutive high expression of the ERG11 gene, coding for the drug target enzyme sterol 14alpha-demethylase, correlated with a stepwise development of fluconazole resistance . The resistant strains exhibited reduced accumulation of fluconazole and, for the last in the series, a slight increase in drug needed to inhibit sterol 14alpha-demethylation in vitro . In the isolates from patient 2, increased MDR1 mRNA levels and the change from heterozygosity to homozygosity for a mutant form of the ERG11 gene correlated with continuously decreased drug susceptibility . In this series, reduced drug accumulation and increased resistance in the target enzyme activity, sterol 14alpha-demethylase, were observed . These results demonstrate that different molecular mechanisms contribute to a gradual development of fluconazole resistance in C . albicans.

Int J Cancer, 1998 Dec 9, 78(6), 740 - 9
Primary human mesothelioma cells express class II MHC, ICAM-1 and B7-2 and can present recall antigens to autologous blood lymphocytes; Mutti L et al.; Mesothelioma cells (MMc) are considered to be weakly immunogenic and the experimental approaches attempting to induce an immune response against these cells have been disappointing . Our aim was to investigate whether MMc possess the surface accessory molecules involved in antigen presentation and whether these cells are capable of presenting recall antigens to autologous blood lymphocytes . Four primary MMc cultures were generated from malignant effusions and examined to assess whether the accessory molecules required for antigen presentation were present on their surfaces . Intercellular adhesion molecule-I (ICAM-I; CD54); class I and class II major histocompatibility complex-DR (MHCI and MHCII-DR); B7-1 (CD80.3); and B7-2 (CD86) expression by MMc was studied by immunocytochemical and/or FACScan analysis . MMc were pulsed with purified protein derivative (PPD), Tetanus toxoid (TT) and Candida albicans (CA) bodies, and incubated with autologous lymphocytes . Lymphocyte proliferation was estimated by radionucleotide incorporation . Phenotypic analysis showed the presence of MHCII-DR, ICAM-I and B7-2 on primary MMc cultures, whereas the phenotypic evaluation of 2 established MMc lines did not show the presence of the B7-1 and B7-2 molecules . In addition, MHCII-DR was detectable only after interferon gamma (IFN-gamma) stimulation . Primary MMc cultures acquired the capability to induce lymphocyte proliferation after pulse with the recall antigens . To achieve characterization of these lymphocytes, we generated a PPD-specific CD4+ T-cell clone . PPD-pulsed MMc were shown to specifically induce T-cell clone proliferation through a MHCII-DR-mediated process . We conclude that primary MMc possess the surface molecules required for antigen presentation and can present recall antigens to CD4+ lymphocytes.

J Acquir Immune Defic Syndr Hum Retrovirol, 1998 Dec 1, 19(4), 373 - 80
Th1/Th2 cytokine expression in saliva of HIV-positive and HIV-negative individuals: a pilot study in HIV-positive individuals with oropharyngeal candidiasis; Leigh JE et al.; Current data suggest that T-helper (Th)2-type cytokine responses are often associated with progression to AIDS in HIV-positive individuals . Similarly, Th2-type cytokines are associated with susceptibility to mucosal candidiasis, of which oropharyngeal candidiasis (OPC) is one of the most common opportunistic infections in HIV-positive individuals . Although little information is available on host defense mechanisms at the level of the oral mucosa, recent studies suggest that local cell-mediated immunity (CMI) is equally or more important than that in the periphery for host defense against mucosal Candida albicans infections . This study investigated the potential presence of oral-associated CMI through the expression of Th1/Th2-type cytokines in saliva of immunocompetent and immunocompromised individuals with and without OPC . Results showed a constitutive mixed Th1/Th2 cytokine expression (Th0) in whole saliva of healthy HIV-negative individuals . In contrast, HIV-positive individuals had a dominant Th2-type salivary cytokine profile (interleukin-4 {IL-4}, IL-10) (IL-2, interferon-y {IFN-gamma}, IL-12) that seemingly resulted from a lack of Th1-type cytokines rather than enhanced Th2-type cytokines . Moreover, pilot analyses of those with OPC showed evidence for a more profound salivary Th2-type profile . Both HIV-positive and HIV-negative patients, irrespective of CD4 counts, had some level of positive in vitro systemic lymphocyte proliferative responses to C albicans antigens . These results suggest that the Th1/Th2 cytokine dichotomy in HIV disease is detectable in situ in oral secretions and may be a useful indicator of oral-associated CMI to better understand resistance/susceptibility of HIV-positive individuals to oral opportunistic infections, including OPC.

J Immunol, 1998 Dec 1, 161(11), 6228 - 37
IL-10 is required for development of protective Th1 responses in IL-12-deficient mice upon Candida albicans infection; Mencacci A et al.; IL-12 is both required and prognostic for Th1 development in mice with Candida albicans infection . To delineate further the physiologic role of IL-12 in antifungal immunity, mice deficient for this cytokine were assessed for susceptibility to C . albicans infections, and for parameters of innate and adaptive immunity . IL-12-deficient mice were highly susceptible to gastrointestinal infection or to reinfection and showed elevated production of Candida-specific IgE and IL-4 and defective production of IFN-gamma . The failure to mount protective Th1 responses occurred despite the presence of an unimpaired innate antifungal immune response, which correlated with unaltered IFN-gamma production, but defective production of, and responsiveness to, inhibitory IL-10 . IL-10 or IL-12 neutralization increased the innate antifungal resistance in wild-type mice . However, in IL-12-deficient mice, treatment with exogenous IL-12 or IL-10 impaired IL-4 production and increased resistance to infection, through a negative effect on the CTLA-4/B7-2 costimulatory pathway . These results confirm the obligatory role of IL-12 in the induction of anticandidal Th1 responses, and indicate the existence of a positive regulatory loop between IL-12 and IL-10 that may adversely affect the innate antifungal response, but is required for optimal costimulation of IL-12-dependent CD4+ Th1 cells.

J Immunol, 1998 Dec 1, 161(11), 6198 - 205
Interaction of the fungal pathogen Candida albicans with integrin CD11b/CD18: recognition by the I domain is modulated by the lectin-like domain and the CD18 subunit; Forsyth CB et al.; Interactions of microorganisms with integrins are central to the host defense mechanisms . The leukocyte integrin CD11b/CD18 is the principal adhesion receptor on leukocytes for Candida albicans, a major opportunistic pathogen . In this study we have investigated the roles of three regions within the receptor, the inserted (I) and lectin-like domains within the CD11b subunit, and the CD18 subunit, in CD11b/CD18-C . albicans interactions . We report four major findings . 1) A mutation in CD18 exerts a dominant negative effect on the function of the CD11b/CD18 complex . This interpretation is based on the observation that in the absence of CD18, the CD11b subunit alone binds C . albicans well, but a single point mutation at Ser138 of CD18 abolishes CD11b/CD18 binding of the fungus . 2) The lectin-like domain is not sufficient for CD11b/CD18-C . albicans interactions . Rather, the lectin-like domain appears to influence CD11b/CD18 binding activity by modulating the function of the I domain . 3) The I domain is the primary binding site for C . albicans in the receptor and is sufficient to support an efficient interaction . 4) We have identified specific amino acid sequences within the I domain that engage the microorganism . Compared with other ligands of CD11b/CD18, C . albicans has some unique as well as common contact sites within the I domain of the receptor . Such unique contact sites may underlie the ability of C . albicans to modulate CD11b/CD18 function and raise the possibility for selective interference of the microorganism-host leukocyte interactions.

Biol Neonate, 1999, 75(1), 31 - 9
Reduced capacity of neonatal lymphocytes to inhibit the growth of Candida albicans; Shareef MJ et al.; Opportunistic microorganisms produce significant morbidity and mortality in preterm and term infants . Because of the heightened susceptibility of infants to opportunistic fungal infections, neonatal lymphocytes were assessed for their capacity to inhibit the growth of Candida albicans . Lymphocytes from both preterm and term cord blood demonstrated significantly less effect upon C . albicans than did lymphocytes from adults . Neonatal lymphocytes of infants <32 weeks of gestation showed a marked reduction in growth inhibitory capacity compared to infants >32 weeks of gestation . Lymphocytes from female infants had a significantly greater fungal growth inhibitory capacity than did lymphocytes from male infants . These results show that neonatal lymphocytes have a reduced capacity to inhibit the growth of C . albicans . This reduced antifungal capacity may underlie the increased susceptibility of such infants to opportunistic microorganisms, like C . albicans.

J Prosthet Dent, 1998 Dec, 80(6), 723 - 9
Effectiveness of chlorine dioxide in disinfection on two soft denture liners; Furukawa KK et al.; STATEMENT OF PROBLEM: Soft tissue denture liners frequently require replacement that necessitates complete removal from the denture base . A high speed lathe located in a "clean laboratory" is often used to facilitate removal of these materials, but it is unclear whether routine disinfection procedures reduce bacterial contamination sufficiently to prevent contamination of the laboratory . PURPOSE: The first phase of this study evaluated the effectiveness of 3-minute chlorine dioxide spray and immersion disinfection procedures on 2 denture liners (Coe Soft and Coe Comfort) and stainless steel specimens used as controls . The second phase evaluated the effectiveness of spray disinfection at time intervals of 1, 3, and 10 minutes . MATERIAL AND METHODS: Specimens made of soft denture liners attached to acrylic resin bases (10 per group) were contaminated with Escherichia coli, Staphylococcus aureus, and Candida albicans . Colony-forming units were counted after different disinfection techniques were applied . Kruskal-Wallis 1-way analysis of variance on ranks and an all pairwise multiple comparison procedures (Dunn's method) were used to test for significant differences among test groups at the P <.05 level of significance . RESULTS: Chlorine dioxide was effective against nonporous stainless steel specimens but was inadequate for denture liners at the recommended 3-minute time of disinfection . The immersion technique was more effective than the spray technique, but the difference was not significant . Increasing the time of disinfection did not significantly reduce the numbers of microorganisms . CONCLUSION: Coe Soft and Coe Comfort denture liners should be removed before entering the laboratory . These materials contain sufficient viable bacteria after routine disinfection procedures to cause contamination of the "clean laboratory."

Clin Infect Dis, 1998 Nov, 27(5), 1291 - 4
Declining rates of oropharyngeal candidiasis and carriage of Candida albicans associated with trends toward reduced rates of carriage of fluconazole-resistant C . albicans in human immunodeficiency virus-infected patients; Martins MD et al.; In order to determine the current prevalence and incidence of fluconazole-resistant oropharyngeal candidiasis among human immunodeficiency virus (HIV)-infected patients, we conducted a prospective observational study of a consecutive series of HIV-infected patients . Of 128 enrolled patients, 70 patients completed four quarterly follow-up visits over a period of 1 year . Over this period, declining rates of carriage of Candida albicans (from 61% to 39%; P = .008) and of oropharyngeal candidiasis (from 30% to 4%; P < .001) were documented . Trends toward reduction in the frequency of fluconazole-resistant isolates (MIC, > or = 64 micrograms/mL) were also seen . During the survey period, the mean (median) number of antiretroviral agents used per patient rose from 0.5 (0) to 1.8 (2) (P < .001) . Thus, rather than progression, we observed declining rates of oropharyngeal candidiasis, C . albicans carriage, and fluconazole-resistant C . albicans in a cohort of HIV-infected patients treated with increasingly effective antiretroviral therapy.

Clin Infect Dis, 1998 Nov, 27(5), 1130 - 3
Candida albicans endophthalmitis in brown heroin addicts: response to early vitrectomy preceded and followed by antifungal therapy; Martinez-Vazquez C et al.; The management of Candida albicans endophthalmitis in intravenous drug abusers (IVDAs) has yet to be established . Early vitrectomy was previously reported as a promising treatment for C . albicans endophthalmitis . In our series, C . albicans endophthalmitis was diagnosed for 15 IVDAs . Funduscopic examinations confirmed severe vitritis in 12 patients and chorioretinitis in three . Blood and vitreal cultures were positive for C . albicans for seven and eight patients, respectively . Patients with vitritis received antifungal therapy before and after vitrectomy . Amphotericin B or fluconazole therapy was given according to the physician's preference . Vitrectomy was defined as early if it was performed within 1 week after the diagnosis of vitritis . All seven patients who underwent early vitrectomy had a favorable response without complications . Two of three patients who underwent late vitrectomy developed blindness or scotoma . Blindness was also described in two patients with vitritis who did not undergo vitrectomy . Early vitrectomy preceded and followed by antifungal therapy seems to be appropriate management of vitritis in IVDAs.

Infect Immun, 1998 Dec, 66(12), 6027 - 9
Contrasting roles of mannan-specific monoclonal immunoglobulin M antibodies in the activation of classical and alternative pathways by Candida albicans; Zhang MX et al.; Polyclonal antimannan immunoglobulin G (IgG) activates the classical complement pathway and accelerates initiation of the alternative pathway by Canidida albicans . This dual role was assessed for two antimannan IgM monoclonal antibodies (MAbs) . MAb B6.1 is specific for an epitope on the acid-labile portion of C . albicans phosphomannan; MAb B6 is specific for an epitope on the acid-stable region . Both MAbs were potent activators of the classical pathway but poor facilitators of alternative pathway initiation.

Infect Immun, 1998 Dec, 66(12), 5812 - 8
Minimum chemical requirements for adhesin activity of the acid-stable part of Candida albicans cell wall phosphomannoprotein complex; Kanbe T et al.; This study was conducted to define adhesive characteristics of the acid-stable moiety of the Candida albicans phosphomannoprotein complex (PMPC) on adherence of this fungus to marginal zone macrophages of the mouse spleen . Complete digestion of the acid-stable moiety (Fr.IIS) of the C . albicans PMPC with an alpha-mannosidase or hydrolysis with 0.6 N sulfuric acid destroyed adhesin activity, as determined by the inability of the soluble digests to inhibit yeast cell adherence to the splenic marginal zone . Fr.IIS adhesin activity was decreased following digestion with an alpha-1,2-specific mannosidase . Oligomannosyls consisting of one to six mannose units, which were isolated from the acid-stable part of the PMPC, did not inhibit yeast cell binding and thus do not function alone as adhesin sites in the PMPC . To gain more insight into the minimum requirements for adhesin activity, PMPCs were isolated from a Saccharomyces cerevisiae wild-type strain and from mutant strains mnn1, mnn2, and mnn4; the PMPCs were designated scwt/Fr.II, scmn1/Fr.II, scmn2/Fr.II, and scmn4/Fr.II, respectively . S . cerevisiae scmn2/Fr.II lacks oligomannosyl side chain branches from the outer core mannan, and scmn2/Fr.II was the only PMPC without adhesin activity . S . cerevisiae scwt/Fr.II, scmn1/Fr.II, and scmn4/Fr.II showed adhesin activities less than that of C . albicans Fr.II . These three S . cerevisiae PMPCs are generally similar to Fr . IIS, except that the S . cerevisiae structure has fewer and shorter side chains . Immunofluorescence microscopy show that the acid-stable part of the PMPC is displayed homogeneously on the C . albicans yeast cell surface, which would be expected for a surface adhesin . Our results indicate that both the mannan core and the oligomannosyl side chains are responsible for the adhesin activity of the acid-stable part of the PMPC.

Infect Immun, 1998 Dec, 66(12), 5771 - 6
A vaccine and monoclonal antibodies that enhance mouse resistance to Candida albicans vaginal infection; Han Y et al.; We previously reported that a vaccine composed of liposome-mannan complexes of Candida albicans (L-mann) stimulates mice to produce protective antibodies against disseminated candidiasis . An immunoglobulin M (IgM) monoclonal antibody (MAb), B6.1, specific for a beta-1,2-mannotriose in the complexes protects against the disease, whereas MAb B6 does not . In the present study, the vaccine and MAbs B6.1 and B6 were tested for the ability to protect against Candida vaginal infection, established by intravaginal (i.vg.) inoculation of yeast cells in mice maintained in pseudoestrus . Fungal CFU in each vagina was determined to assess the severity of infection . Mice vaccinated before infection developed about 62% fewer vaginal CFU than nonimmunized controls . Naive mice that received polyclonal antiserum (from vaccinated mice) i.vg . before infection had 60% fewer CFU than controls . The serum protective factor was stable at 56 degreesC, but C . albicans cells absorbed this factor . Mice given MAb B6.1 i.vg . after infection was established had fewer Candida CFU in vaginal tissue than control mice given buffer instead of antibody . MAbs B6.1 and B6 given intraperitoneally before infection protected mice, but MAbs preabsorbed with yeast cells did not . MAb B6.1 also protected against C . tropicalis vaginal infection, but MAb B6 did not . The protective activities of MAbs B6.1 and B6 appeared to be specific because an irrelevant IgM carbohydrate-specific MAb and an irrelevant IgG protein-specific MAb were not protective; also, MAb B6.1 did not affect development of vaginal chlamydial infection . These studies show that an appropriate antibody response, or administration of protective antibodies, can help the host to resist Candida vaginal infection.

Photochem Photobiol, 1998 Nov, 68(5), 738 - 44
Suppression of delayed and contact hypersensitivity responses in mice have different UV dose responses; Kim TH et al.; Although acute exposure to UV radiation suppresses the induction of delayed-type (DTH) and contact (CHS) hypersensitivity in mice, it is not clear whether the photobiological mechanism(s) involved in suppressing these closely related immune reactions is the same . A careful examination of the UV dose responses and wavelength dependencies involved in suppressing CHS and DTH may provide important insights into the mechanisms involved . We compared the UV dose-response curves for suppressing four closely related immune reactions, local and systemic suppression of CHS to dinitrofluorobenzene, systemic suppression of DTH to Candida albicans and systemic suppression of DTH to alloantigen using three different UV spectra (FS40 sunlamps, Kodacel-filtered FS40 sunlamps and solar-simulated light) . For each immune response studied, the amount of UVB radiation required to induce 50% immune suppression was lowest when FS40 sunlamps were used, highest with solar-simulated light and intermediate when Kodacel-filtered FS40 sunlamps were used, but the differences observed were not statistically significant . The UV dose-response curves for immune suppression differed significantly depending on the assay used, the site of antigenic sensitization and the antigen used . These findings suggest that the mechanisms by which UV radiation induces immune suppression differ for the four immunological reactions studied.

J Leukoc Biol, 1998 Nov, 64(5), 650 - 6
Glycosaminoglycan profile in macrophages exposed to Candida albicans and interleukins; Bodo M et al.; Glycosaminoglycans (GAG), are extracellular matrix macromolecules that affect the phagocytic properties of macrophages . In order to assess whether the interaction between macrophages and Candida albicans (iCa) provokes changes in the phenotype, we analyzed the GAG profiles in two macrophage lines, ANA-1 (from murine bone-marrow) and BV-2 (from murine brain) . We also investigated GAG modulation by interleukin-1alpha (IL-1alpha) and interleukin-6 (IL-6) . During iCa treatment and even after the addition of ILs, ANA-1 accumulated less total GAG compared to controls . IL-1 treatment, combined with iCa exposure, induced a decrease in heparan sulfate and chondroitin sulfate chains, and an increase in the hyaluronic acid percentage . IL-6 treatment, with or without iCa, decreased the hyaluronic acid/sulfated GAG ratio . The GAG pattern in BV-2 appears to be different to ANA-1 and iCa exposure does not induce any difference in total GAG . The inhibitory effect induced by ILs on GAG synthesis is less than that observed in ANA-1 and the GAG elution profile is modulated to a lesser extent by treatment with ILs and/or iCa compared to the ANA-1 . We suggest that the observed changes in the expression of the individual GAG classes may be responsible for the macrophage functional heterogeneity.

J Med Microbiol, 1998 Nov, 47(11), 1007 - 14
Antifungal activity of interleukin-2-activated natural killer (NK1.1+) lymphocytes against Candida albicans; Mathews HL et al.; Interleukin-2 (IL-2)-activated lymphocytes interact directly with, and inhibit, the growth of Candida albicans hyphae . C . albicans-stimulated natural killer (NK1.1+) lymphocytes were demonstrated to secrete a soluble product capable of directly affecting C . albicans yeast forms . Antibodies specific for interferon-gamma completely eliminated the antifungal activity of the NK1.1+ lymphocyte product and diminished the antifungal activity of NK1.1+ lymphocytes against C . albicans . Antibodies specific for other cytokines had no such effect . These data demonstrate that C . albicans-stimulated NK1.1+ lymphocytes have antifungal activity against C . albicans yeast cells via the release of interferon-gamma . This antifungal activity was demonstrable only against the yeast form of the fungus, with no effect on C . albicans hyphae.

Arch Oral Biol, 1998 Nov, 43(11), 879 - 87
The effect of limited exposure to antimycotics on the relative cell-surface hydrophobicity and the adhesion of oral Candida albicans to buccal epithelial cells; Ellepola AN et al.; Candida albicans is the major aetiological agent of oral candidosis . Adhesion to oral mucosal surfaces is considered a prerequisite for its successful colonization and subsequent infection, and its relative cell-surface hydrophobicity (CSH) is a contributory physical force . Thus, the main aim here was to determine the CSH of 10 isolates of oral C . albicans after a short exposure to sublethal concentrations of four antifungal agents and to correlate these findings with their adhesion to human buccal epithelial cells (BEC) . The yeasts were exposed to sublethal concentrations of nystatin {x 6 minimal inhibitory concentration (MIC)}, 5-fluorocytosine (x 8 MIC), ketoconazole (x 4 MIC) and fluconazole (x 4 MIC) for 1 h . The drug was then removed, and the CSH and BEC adhesion assessed by a biphasic aqueous-hydrocarbon assay and a microscopic method, respectively . The mean percentage reductions of CSH after exposure to nystatin, 5-fluorocytosine, ketoconazole and fluconazole were 27.14% (p = 0.01), 9.46% (p = 0.43), 19.47% (p = 0.04) and 6.16% (p = 0.59) . Similarly, exposure to all the drugs except 5-fluorocytosine resulted in a significant inhibition of yeast adhesion to BEC, with nystatin eliciting the highest and fluconazole the least inhibition . Further, on regression analysis a strong positive correlation was observed between CSH and adhesion to BEC after limited exposure to 5-fluorocytosine (r = 0.48, p < 0.0001), ketoconazole (r = 0.48, p < 0.0001), fluconazole (r = 0.55, p < 0.0001) as well as in the unexposed controls (r = 0.41, p = 0.001), although nystatin was an exception (r = 0.09, p = 0.44) . Taken together, these data elucidate further mechanisms by which antimycotics may operate in vivo to suppress candidal pathogenicity.

J Infect, 1998 May, 36(3), 259 - 67
A model of human cutaneous candidosis based on reconstructed human epidermis for the light and electron microscopic study of pathogenesis and treatment; Korting HC et al.; The use of reconstructed human epidermis provided the basis for an in vitro model of human cutaneous candidosis . Candida albicans blastospores on the surface of reconstructed human epidermis provoked the following changes within 72 h: superficial keratin degradation, scaling, hyperkeratosis, parakeratosis, dyskeratosis representing hyperproliferative stress, spongiosis, and vesiculation . Great differences in the intensity of these reactions of intact reconstructed human epidermis or chemically or mechanically damaged reconstructed human epidermis illustrate the importance of the stratum corneum as a barrier . Uninfected reconstructed human epidermis showed prominent cell proliferation representing wound healing 72 h after mechanical or chemical pretreatment . These signs of repair were blocked in the presence of C . albicans and the blastospores were able to invade the stratum corneum . When desmosomes were accessible, a high affinity of C . albicans blastospores to these structures was observed . A single application of an econazole liposome dispersion decreased scaling, hyperkeratosis and dyskeratosis . Morphological alterations of C . albicans blastospores after treatment with the econazole liposome dispersion in the proposed ill vitro model were identical, as described in established animal models . This reconstructed human epidermis model of cutaneous disease may provide insight into the pathogenesis and treatment of cutaneous candidosis and may provide a substitute for animal models and investigations on humans.

J Infect, 1998 Jan, 36(1), 57 - 62
Variation in morphotype, karyotype and DNA type of fluconazole resistant Candida albicans from an AIDS patient; Takasuka T et al.; Azole-resistant oropharyngeal and oesophageal candidiasis is a recent phenomenon observed in patients with AIDS usually previously treated with fluconazole . Some variation has been observed in antifungal susceptibility testing among separate colonies of Candida albicans from the same patient . This raises the question of whether there are multiple clones present or simply phenotypic variation in expression of azole resistance . To address this question we took 18 isolates grown from multiple swabs taken before and after experimental azole therapy from a single HIV-positive individual with fluconazole-resistant oral candidiasis and compared morphotype, karyotype, PCR-based DNA typing and azole susceptibility . Ten of the isolates were from a single 2-day period . Amongst these 10 there were seven morphotypes, five karyotypes and four polymerase chain reaction (PCR) types . Three further morphotypes, one karyotype and two PCR types were found amongst the eight isolates obtained during the subsequent 4 months . Limited variation in susceptibility to two azoles--fluconazole and D0870--was also seen . This work emphasizes both the large genotype and phenotypic variability of C . albicans isolates in the mouth of AIDS patients with fluconazole resistance, and the difficulties in interpretation of present typing methods.

Microb Drug Resist, 1998 Fall, 4(3), 143 - 58
In vivo characterization of the drug resistance profile of the major ABC transporters and other components of the yeast pleiotropic drug resistance network; Kolaczkowski M et al.; Multidrug resistance (MDR) mediated by broad specificity transporters is one of the most important strategies used by pathogens, including cancer cells, to evade chemotherapy . In the yeast Saccharomyces cerevisiae, a complex pleiotropic drug resistance (PDR) network of genes involved in MDR is composed of the transcriptional regulators Pdr1p and Pdr3p, which activate expression of the ATP-binding cassette (ABC) MDR transporters-encoding genes PDR5, SNQ2, and YOR1 as well as other not yet identified genes . We have screened 349 toxic compounds in isogenic S . cerevisiae strains deleted of PDRS, SNQ2, or YOR1 in different combinations as well as both PDR1 and PDR3 . The screen revealed extremely promiscuous, yet limited, and to a large extent overlapping but distinct drug resistance profiles of Pdr5p, Snq2p, and Yor1p . These ABC-MDR transporters mediated resistance to most currently available classes of clinically and agriculturally important fungicides and also to many antibiotics, herbicides, and others . Several classes of compounds were identified for the first time in the drug resistance spectrum of MDR transporters . These are fungicides, such as anilinopyrimidines, benzimidazoles, benzenedicarbonitriles, dithiocarbamates, guanidines, imidothiazoles, polyenes, pyrimidynyl carbinols, and strobilurine analogues; the urea derivative and anilide herbicides; flavonoids, several membrane lipids resembling detergents; and newly synthesized lysosomotropic aminoesters; as well as many others . Identification of compounds showing Pdr1p, Pdr3p-dependent, but Pdr5p-, Snq2p-, and Yor1p-independent toxicity, reflected in the case of rhodamine 6G, by efflux alterations, suggests the involvement of new drug resistance genes and is a first step toward their identification . The highly increased toxicity of bile acids toward the PDR1, PDR3 double disruptant together with the decreased level of BAT1 promoter dependent beta-galactosidase activity suggest that the Bat1p ABC transporter is a new member of the PDR network . Our results may contribute to a better understanding of the mechanism of MDR, in particular in the pathogenic yeast Candida albicans . They also provide and indication of the physiological function of MDR transporters and suggest new approaches for the cloning of the mammalian bile acid transporters.

J Int Med Res, 1998 Aug-Sep, 26(4), 209 - 18
Fluconazole orally dispersible tablets for the treatment of patients with oropharyngeal candidiasis; Vandercam B et al.; The efficacy and tolerability of fluconazole orally dispersible tablets (ODT) in the treatment of oropharyngeal candidiasis was evaluated in this multicentre non-comparative study . A total of 89 adults with signs and symptoms of oropharyngeal candidiasis were enrolled; 70 of whom completed therapy with fluconazole ODT 100 mg once daily for 7 - 14 days . Acquired immunodeficiency syndrome (AIDS)/ AIDS-related complex was an underlying illness in 69% of patients (61) . An antimicrobial and corticosteroid therapy was given in 52% (46) and 20% (18) of patients, respectively . Of the 60 patients who had baseline signs and symptoms of infection and a culture positive for Candida albicans, 90% (54) were cured or had improved at the end of therapy, and the fungal pathogen was eradicated in 19/57 (33%) patients . At the 4-week posttreatment follow-up, signs and symptoms of oropharyngeal candidiasis were absent in 73% (27/37) patients . The adverse events and laboratory abnormalities recorded during the study period were attributable to underlying illnesses rather than to fluconazole therapy . These results indicate that this novel dosage form of fluconazole is effective and well tolerated in the treatment of oropharyngeal candidiasis.

J Antimicrob Chemother, 1998 Oct, 42(4), 469 - 74
Modulation of the pro- and anti-inflammatory cytokine balance by amphotericin B; Vonk AG et al.; Amphotericin B is an antifungal drug associated with side effects such as fever and chills, symptoms which may be mediated by pro-inflammatory cytokines such as interleukin-1beta (IL-1beta) and tumour necrosis factor-alpha (TNFalpha) . We assessed the capacity of amphotericin B to modulate production of these pro-inflammatory cytokines as well as the anti-inflammatory IL-1 receptor antagonist (IL-1ra), induced by LPS, heat-killed Candida albicans or Staphylococcus aureus . The results of the present study show that amphotericin B slightly increased the production of pro-inflammatory cytokines by human mononuclear cells (PBMC), whereas the production of the anti-inflammatory cytokine IL-1ra was significantly inhibited . This results in a shift towards pro-inflammatory cytokine production, as indicated by a decreased IL-1ra/IL-1beta ratio . Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) indicated that levels of IL-1beta and TNFalpha mRNA were increased . In conclusion, amphotericin B is able to cause a shift towards pro-inflammatory cytokine production by human PBMC . This may explain the side effects, such as fever and chills, observed after treatment of patients with amphotericin B.

J Infect Dis, 1998 Dec, 178(6), 1743 - 9
Serum-mediated enhancement of TNF-alpha release by human monocytes stimulated with the yeast form of Candida albicans; Ghezzi MC et al.; The role of serum in the production of tumor necrosis factor-alpha (TNF-alpha) by human monocytes stimulated with yeast-form Candida albicans was studied . Pre-exposure of C . albicans to human pooled serum enhanced both TNF-alpha mRNA and cytokine secretion compared with C . albicans preincubated with medium only . Serum factors involved were >30 kDa, were efficiently inhibited by D-mannose, and recognized both Ca++-dependent and -independent pathways . Preincubation of yeasts with rabbit mannose-binding protein (MBP) resulted in dose-related enhancement of TNF-alpha secretion, through a Ca++-dependent pathway inhibited by D-mannose . TNF-alpha levels were similarly induced in C . albicans preincubated with vitronectin and with serum . Ca++ depletion did not affect cytokine release, while D-mannose supplementation displayed inhibition . The latter effect was abolished after Ca++ depletion . These data call for an involvement of both MBP and vitronectin in the serum-mediated enhancement of TNF-alpha release upon stimulation of monocytes with yeast forms of C . albicans.

J Infect Dis, 1998 Dec, 178(6), 1734 - 42
Suppressive effects of interleukin-10 on human mononuclear phagocyte function against Candida albicans and Staphylococcus aureus; Roilides E et al.; The effects of interleukin (IL)-10, a potent antiinflammatory cytokine, on human monocyte functions against two medically important pathogens, Candida albicans and Staphylococcus aureus, were studied . Incubation with 20-100 ng/mL IL-10 for 2-3 days decreased the fungicidal activity of monocytes against serum-opsonized C . albicans blastoconidia (P</=.04), reduced their capacity to damage unopsonized hyphae (P</=.006), and suppressed superoxide anion production in response to phorbol myristate acetate (P=.019) and N-FMLP (P=.04) but not to serum-opsonized blastoconidia . Paradoxically, IL-10 enhanced phagocytic activity of monocytes against serum-opsonized blastoconidia (P<.01) . In addition, IL-10-treated monocytes demonstrated decreased bactericidal activity (P=.046) but no change in bacterial phagocytosis . These findings demonstrate an overall suppressive role of IL-10 on human monocyte function against C . albicans and S . aureus and may have important implications in the use of this cytokine.

Infect Dis Obstet Gynecol, 1998, 6(4), 176 - 81
17-beta-estradiol upregulates the stress response in Candida albicans: implications for microbial virulence; O'Connor C et al.; OBJECTIVE: The influence of 17-beta-estradiol on the stress response of Candida albicans was studied . METHODS: The survival of clinical isolates of C . albicans treated with 17-beta-estradiol after heat and oxidative stress was measured by viable plate counts . Cellular proteins were analyzed via SDS-PAGE . RESULTS: The heat stress response induced by 17-beta-estradiol in C . albicans grown at 25 degrees C protected the organisms against the lethal temperature of 48.5 degrees C, as shown by viable plate counts . 17-beta-estradiol also enhanced protection of C . albicans against oxidative stress (menadione exposure) . SDS-PAGE analysis of cytoplasmic extracts revealed proteins induced by 17-beta-estradiol were similar to those induced by heat . CONCLUSION: 17-beta-estradiol enhances survival of C . albicans under heat and oxidative stresses . The proteins induced by 17-beta-estradiol are probably heat shock proteins . Because heat shock proteins are considered to be virulence factors, 17-beta-estradiol may function to promote in vivo survival.

Infect Dis Obstet Gynecol, 1998, 6(4), 168 - 75
Effect of gliotoxin on human polymorphonuclear neutrophils; Shah DT et al.; OBJECTIVES: Candida albicans is known to produce gliotoxin, which has several prominent biological effects, including immunosuppression . Interference with host defenses may arise from the effects of this toxin on leukocyte structure and function . METHODS: Flow cytometric analysis revealed that polymorphonuclear leukocytes (PMN) were more sensitive to gliotoxin than were mononuclear cells . Structural and various functional aspects of PMN exposed to gliotoxin were studied . RESULTS: Gliotoxin at (1 microgram/mL) did not affect the viability but did diminish PMN chemotaxis and reduced their ability to ingest particles . Other functional aberrations included decreased nitroblue tetrazolium dye reduction, decreased superoxide production, and release of lactoferrin suggesting by degranulation . Gliotoxin also affected the ability of PMN to kill Escherichia coli . CONCLUSIONS: This study suggests a previously unrecognized potential virulence factor of C . albicans that could contribute to persistence of yeast colonization or recurrence of symptomatic infection through diminished host resistance.

Neurologia, 1998 Aug-Sep, 13(7), 362 - 6
Chronic neutrophilic meningitis caused by Candida albicans; del Pozo MM et al.; Meningitis caused by Candida albicans is a very infrequent entity . We present 3 intravenous drug users with chronic neutrophilic meningitis caused by Candida albicans . The clinical, microbiological and radiological features of the 3 patients are reviewed . The interval between the onset of the disease and the diagnosis was long (from 4 to 12 months) . Candida albicans was cultured from cerebrospinal fluid (CSF) in the 3 patients . All of them developed hydrocephalus, meanwhile arachnoiditis was disclosed in two . The therapy with amphotericyn, 5-flucytosine and fluconazole produced clinical improvement and the sterilisation of the CSF in all the 3 cases . Clinicians should be aware of this entity because of diagnosis may be delayed in several months.

FEMS Microbiol Lett, 1998 Oct 15, 167(2), 163 - 9
Cloning and regulated expression of the Candida albicans phospholipase B (PLB1) gene; Hoover CI et al.; Degenerate oligonucleotides (derived from conserved regions of PLB1 genes from S . cerevisiae and other fungi) were used to amplify PLB1 homolog fragments from C . albicans and C . tropicalis by using the polymerase chain reaction . The C . albicans PLB1 fragment was then used as a probe to clone the full-length gene and to monitor PLB1 mRNA expression . The C . albicans PLB1 gene consists of a 1815-bp open reading frame encoding a putative protein of 605 amino acids . It contains the highly conserved Gly-X-Ser-X-Gly catalytic motif, found in all lipolytic enzymes, and exhibits significant homology with other fungal PLB1 gene products (approximately 63% similarity, approximately 45% identity) . Blastospores and pseudohyphae expressed higher levels of PLB1 mRNA than germ-tube-forming cells . TUP1, a general transcriptional repressor, may regulate PLB1 expression in C . albicans, since PLB1 expression was the highest in tup1 delta mutants and did not vary in response to environmental stimuli . Together, these results suggest that expression of the C . albicans PLB1 gene is regulated as a function of morphogenic transition.

Clin Rheumatol, 1998, 17(5), 393 - 4
Candida arthritis in an immunocompetent patient without predisposing factors; Calvo Romero JM et al.; Candida arthritis is infrequent in patients who are not intravenous drug users . We describe a patient who was not an intravenous drug user and without other predisposing factors who developed knee arthritis caused by Candida albicans . We believe that this is the first report of such a case.

Yeast, 1998 Oct, 14(14), 1327 - 32
Single-read sequence tags of a limited number of genomic DNA fragments provide an inexpensive tool for comparative genome analysis; Hartung K et al.; Single-read sequences from both ends of 415 3-kb average size genomic DNA fragments of Candida albicans were compared with the complete sequence data of Saccharomyces cerevisiae . Comparison at the protein level, translated DNA against protein sequences, revealed 138 sequence tags with clear similarity to S . cerevisiae proteins or open reading frames . One case of synteny was found for the open reading frames of RAD16 and LYS2, which are adjacent to each other in S . cerevisiae and C . albicans.

Yeast, 1998 Oct, 14(14), 1257 - 65
Cloning of the Candida albicans nucleoside transporter by complementation of nucleoside transport-deficient Saccharomyces; Detke S; The nucleoside permease gene (i.e . NUP) from Candida albicans was cloned by complementation of Saccharomyces cerevisiae deficient in nucleoside transport capability . The permease transported adenosine and guanosine and was sensitive to the mammalian nucleoside transport inhibitors: dipyridamole and NBMPR . It did not transport uridine, cytidine, adenine, guanine or uracil . The inability to transport uridine indicated that the NUP gene product was different from the Candida uridine permease, which also transported cytosine and adenosine . The NUP gene coded for a protein of 407 amino acids in size which was approximately the size of the human, Giardia and E . coli nucleoside permeases . It did not, however, exhibit any significant degree of homology with these transporters.

Microbiology, 1998 Oct, 144 ( Pt 10), 2731 - 7
Differential regulation of SAP8 and SAP9, which encode two new members of the secreted aspartic proteinase family in Candida albicans; Monod M et al.; Secreted aspartic proteinases (Saps) contribute to the virulence of Candida albicans in systemic animal models of infection . Seven genes encoding Saps (SAP1-SAP7) have been identified to date but evidence suggested the existence of additional SAP genes . The screening of a C . albicans lambda EMBL3 genomic library for the presence of other SAP genes was undertaken . Two new genes, SAP8 and SAP9, were isolated . The N-terminal amino acid sequence deduced from SAP8 downstream of a Kex2p-like cleavage site corresponds to the N-terminal amino acid sequence of the 41 kDa Sap isolated and characterized previously . SAP8 mRNA was expressed preferentially in yeasts at 25 degrees C after 6 and 9 h growth in BSA-containing medium . SAP9 encodes an aspartic proteinase with a Kex2p-like cleavage site and contains a putative glycophosphatidylinositol-anchor signal at the C-terminus . Although the SAP9 gene product has not yet been isolated from cultures of C . albicans, transcripts of SAP9 were observed preferentially in later growth phases when SAP8 expression had decreased.

Microbiology, 1998 Oct, 144 ( Pt 10), 2715 - 29
The two-component hybrid kinase regulator CaNIK1 of Candida albicans; Srikantha T et al.; Using degenerate primers of highly conserved regions of two-component response regulators for PCR amplification, a two-component response regulator was cloned from Candida albicans that is homologous to nik-1+ of Neurospora crassa . This two-component hybrid kinase, CaNIK1, also shows features of bacterial two-component response regulators, including a putative unorthodox second histidine kinase motif at the carboxy-terminal end . CaNIK1 was expressed at low levels in both the white and opaque switch phenotypes and in the bud and hyphal growth forms of C . albicans strain WO-1, but in both developmental programmes, the level of transcript was modulated (levels were higher in opaque cells and in hyphae) . Partial deletion of both CaNIK1 alleles, by which the histidine autokinase- and ATP-binding domains were removed, did not inhibit either high-frequency phenotypic switching or the bud-hypha transition in high salt concentrations, but in both cases the efficiency of the developmental process was reduced.

Nucleic Acids Res, 1998 Nov 15, 26(22), 5061 - 6
Overlapping coding regions and trancriptional units of two essential chromosomal genes (CCT8, TRP1)in the fungal pathogen Candida albicans; Gerads M et al.; Sequencing of the 3'-untranslated region of the CCT8 gene of the fungal pathogen Candida albicans revealed that the CCT8 coding region overlaps 13 bp with the coding region of the convergently orientated TRP1 gene . The same overlap was found in three strains with different genetic backgrounds . 3'-RACE was used to determine that the CCT8 and TRP1 transcripts extended significantly into the coding region of the adjacent gene, which also contained sequences encoding the poly(A) addition site . A strain retaining one wild-type CCT8/TRP1 locus on one chromosome and a deletion on the other homologous chromosome contained both CCT8 and TRP1 transcripts; this result indicates that both transcripts are synthesized from the same gene locus . The CCT8/TRP1 gene pair of C . albicans constitutes an extreme natural case of transcriptional overlap in a eukaryote . The results confirm that convergent overlapping transcription units are compatible with expression of the overlapping genes.

Clin Infect Dis, 1998 Oct, 27(4), 688 - 91
Candida albicans endocarditis associated with a contaminated aortic valve allograft: implications for regulation of allograft processing; Kuehnert MJ et al.; A patient developed Candida albicans endocarditis and fungemia after undergoing aortic valve replacement with an allograft . The allograft had been found during tissue bank processing to be contaminated with C . albicans, but it was culture-negative for C . albicans after routine disinfection with an antifungal-containing antimicrobial solution . Comparison of the preimplantation and postimplantation C . albicans isolates revealed remarkable genetic similarity, but antifungal susceptibility testing showed that the postimplantation isolate was more resistant to fluconazole and amphotericin B than the preimplantation isolate, suggesting emergence of resistance after disinfection . Implantation of a contaminated heart valve allograft can occur despite disinfection during processing and can result in endocarditis in the recipient . Antimicrobial disinfection protocols that include antifungal drugs may be ineffective . Current U.S . Food and Drug Administration regulations do not require companies to specify details concerning allograft processing . Additional measures may be required to prevent tissue bank release of allografts contaminated with C . albicans or other pathogens.

Transplantation, 1998 Oct 15, 66(7), 828 - 31
1,25-Dihydroxyvitamin D3 prolongs graft survival without compromising host resistance to infection or bone mineral density; Cantorna MT et al.; BACKGROUND: Recently, we have shown that 1,25-dihydroxyvitamin D3 prolongs graft survival in mice and rats when the donor and recipient differ at two or more major histocompatability loci . Among the most serious side effects encountered with the currently available transplantation antirejection drugs are an increased susceptibility to infection and decreased bone mineralization . Our results suggest that 1,25-dihydroxyvitamin D3 prolongs graft survival without these side effects of bone loss and susceptibility to infection . METHODS: We compared the ability of 1,25-dihydroxyvitamin D3-treated, nontreated, or cyclosporine (CsA)-treated mice to resist infection with Candida albicans and herpes simplex virus-1 . To determine bone density, femurs were collected from nontreated, 1,25-dihydroxyvitamin D3-treated (50 ng/mouse/day), or CsA-treated (25 mg/kg/day) mice, and bone ash was determined . RESULTS: Here we show that 1,25-dihydroxyvitamin D3 treatment does not increase the susceptibility of the host to fungal or viral infection . Furthermore, CsA causes bone loss, whereas 1,25-dihydroxyvitamin D3 actually increases bone mass . CONCLUSIONS: The use of 1,25-dihydroxyvitamin D3 and its analogs to increase transplant survival will avoid bone loss and opportunistic infection, two important disadvantages of the most widely used transplant antirejection drugs--CsA and the glucocorticoids.

Antimicrob Agents Chemother, 1998 Nov, 42(11), 3018 - 21
Organism-dependent fungicidal activities of azoles; Manavathu EK et al.; We investigated the antifungal activities of itraconazole and voriconazole on Aspergillus species by time kill studies, and the results were compared with those obtained for Candida species . Exposure of Aspergillus fumigatus conidia to varying concentrations (1.25 to 10 microg/ml) of itraconazole and voriconazole resulted in cellular death; the cytocidal effect was time and concentration dependent . In contrast, no killing of Candida albicans occurred in the presence of itraconazole and voriconazole at concentrations as high as 10 microg/ml, although candidal growth was inhibited compared to the drug-free control . Amphotericin B (1.25 to 10 microg/ml), on the other hand, killed both A . fumigatus and C . albicans . Similar results were obtained for non-A . fumigatus aspergilli and non-C . albicans Candida species . These observations indicate that both itraconazole and voriconazole are cytocidal agents for Aspergillus species but not for Candida species, suggesting that azoles possess organism-dependent fungicidal activities.

Antimicrob Agents Chemother, 1998 Nov, 42(11), 2938 - 42
Fluconazole versus Candida albicans: a complex relationship; Graybill JR et al.; A murine model of systemic candidiasis was used to assess the virulence of serial Candida albicans strains for which fluconazole MICs were increasing . Serial isolates from five patients with 17 episodes of oropharyngeal candidiasis were evaluated . The MICs for these isolates exhibited at least an eightfold progressive increase from susceptible (MIC < 8 microg/ml; range, 0.25 to 4 microg/ml) to resistant (MIC >/= 16 microg/ml; range, 16 to >/=128 microg/ml) . Virulence of the serial isolates from three of five patients showed a more than fivefold progressive decrease in the dose accounting for 50% mortality and was associated with development of fluconazole resistance . Low doses of fluconazole prolonged survival of mice infected with susceptible yeasts but failed to prolong survival following challenge with a resistant strain . In addition, a decreased burden of renal infection was noted in mice challenged with two of the three resistant strains . This was consistent with reduced virulence . Fluconazole did not further decrease the level of infection . In the isolates with a decrease in virulence, two exhibited overexpression of CDR, which encodes an ABC drug efflux pump . In contrast, serial isolates from the remaining two patients with the development of resistance did not demonstrate a change in virulence and fluconazole remained effective in prolonging survival, although significantly higher doses of fluconazole were required for efficacy . Resistant isolates from both of these patients exhibited overexpression of MDR . This study demonstrates that decreased virulence of serial C . albicans isolates is associated with increasing fluconazole MICs in some cases but not in others and shows that these low-virulence strains may not consistently cause infection.

Antimicrob Agents Chemother, 1998 Nov, 42(11), 2932 - 7
Distinct patterns of gene expression associated with development of fluconazole resistance in serial candida albicans isolates from human immunodeficiency virus-infected patients with oropharyngeal candidiasis; Lopez-Ribot JL et al.; Resistance to fluconazole is becoming an increasing problem in the management of oropharyngeal candidiasis in human immunodeficiency virus-infected patients . Strains obtained from five patients developed decreased fluconazole susceptibility over time . DNA strain typing confirmed the high degree of relatedness among isolates from one patient and the variability among isolates from different patients . Expression of genes involved in development of fluconazole resistance was monitored in each isolate using probes specific for ERG11 (lanosterol 14alpha-demethylase), MDR1 (a major facilitator), and CDR (ATP-binding cassette or ABC transporter) genes . Increased expression of CDR genes was detected in the series of isolates from two patients . Isolates from one of the two patients also demonstrated increased ERG11 expression, whereas isolates from the other patient did not . Increased levels of MDR1 mRNA correlated with increased resistance in sequential isolates from another patient . Initial overexpression of MDR1 with subsequent overexpression of CDR genes and a final isolate again overexpressing MDR1 were detected in serial isolates from another patient . In another patient, overexpression of these genes was not detected despite an eightfold increase in fluconazole MIC . In this patient, sequence data of the ERG11 gene revealed no point mutations associated with decreased susceptibility . Five different patterns of gene expression were observed in isolates recovered from five patients who developed resistance . Therefore, these experiments demonstrate that a variety of mechanisms or combinations of mechanisms are associated with the development of fluconazole drug resistance . Additional studies are needed to estimate the frequency and clinical impact of these mechanisms of resistance.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(4), 229 - 33
Analysis by pulsed-field gel electrophoresis of Candida albicans that developed resistance during antifungal therapy; Mori T et al.; A patient with myelofibrosis complicated by recurrent candidemia died despite treatment with amphotericin B and fluconazole . Autopsy revealed systemic candidiasis with fungal verrucae in the right ventricle and the root of the pulmonary artery . The strains of Candida albicans isolated from the blood had become resistant to amphotericin B and fluconazole during therapy, as well as to other azole antifungals that had not been used . Pulsed-field gel electrophoresis showed that the resistant isolates had the same genotype as the sensitive strains isolated before treatment, but a chromosomal change in >2.0 Mb-bands was observed after treatment . It was thus proved that these repeatedly isolated C . albicans strains which were causing the continued fungemia in our patient were all the same strain and were acquiring resistance to antifungal agents during the therapy.

Obstet Gynecol, 1998 Nov, 92(5), 757 - 65
Vulvovaginal candidiasis: clinical manifestations, risk factors, management algorithm; Eckert LO et al.; OBJECTIVE: To correlate symptoms, signs, and risk factors with positive wet mounts or cultures for Candida albicans and to develop an algorithm to diagnose vulvovaginal candidiasis . METHODS: This cross-sectional study of 774 randomly selected women from an urban sexually transmitted disease (STD) clinic evaluated symptoms, signs, and risk factors associated with C albicans, detected by wet mount and culture, and constructed an algorithm . RESULTS: C albicans, recovered from 186 (24%) of the 774 women, was associated with chief complaints of vulvar pruritus or burning . Elicited symptoms were vulvar pruritus, pain or burning, and external dysuria; signs were vulvar erythema, edema, fissures, vaginal erythema, and thick, curdy vaginal discharge . Among 545 women with symptoms of either increased vaginal discharge or vulvar pruritus or burning, only 155 (28%) had positive C albicans cultures, whereas bacterial vaginosis or other sexually transmitted infections were found in 288 (53%) . In multivariate analysis, risk factors for positive C albicans culture included condom use, presentation after the 14th menstrual cycle day, sexual intercourse more than four times per month, recent antibiotic use, young age, past gonococcal infection, and absence of current gonorrhea or bacterial vaginosis . A clinical algorithm based on symptoms, signs, and selective use of wet mounts and cultures would have provided prompt treatment to 150 of 167 (90%) women with vulvovaginal candidiasis while minimizing the number of cultures performed . CONCLUSION: A simple algorithm using symptoms, signs, wet mounts, and selective cultures can identify 90% of women with vulvovaginal candidiasis . In this STD clinic, vulvovaginal symptoms also require assessment for bacterial vaginosis, trichomoniasis, and cervical infection.

Minerva Stomatol, 1998 Jul-Aug, 47(7-8), 293 - 7
{Candida albicans morphotypes and in vitro adhesivity to cells of the human oral mucosa in HIV-positive and AIDS patients after exposure of blastospores to fluconazole . II}; Spadari E et al.; BACKGROUND: Oro-pharyngeal candidosis is a frequently initial clinical manifestation of HIV infection and the adhesive properties of Candida spp . represent a very important pathogenicity factor . METHODS: In this study the adhesivity rate of Candida albicans to the oral epithelial cells of 33 HIV-positive patients and 12 healthy volunteers, have been assessed before and after the exposure of blastospores to inhibitory concentrations of fluconazole, in relation to 11 morphotypes obtained from 13 C . albicans strains . RESULTS: Results can be summarized as follows: 1) the number of blastospores adhering to the HIV-positive donor' cells is higher than that of blastospores adhering to the healthy donors' cells (rate is 2.7:1); 2) blastospores from strains producing rough or very coarse fringes show adhesive properties higher than those of strains with different morphology; 3) in the group of HIV-positive patients the adhesivity inhibition of blastospores from strains producing rough or very coarse fringes was higher (38.3%) than that of strains with different morphology (33.8%); 4) overall, adhesivity inhibition due to exposure to fluconazole is higher for epithelial cells from healthy donors . CONCLUSIONS: These results can suggest the validity of an antimycotic pretreatment of persons at risk of oro-pharyngeal candidiasis.

Yeast, 1998 Sep 30, 14(13), 1159 - 66
A spheroplast rate assay for determination of cell wall integrity in yeast; Ovalle R et al.; The rate of formation of spheroplasts of yeast can be used as an assay to study the structural integrity of cell walls . Lysis can be measured spectrophotometrically in hypotonic solution in the presence of Zymolyase, a mixture of cell wall-digesting enzymes . The optical density of the cell suspension decreases as the cells lyse . We optimized this assay with respect to enzyme concentration, temperature, pH, and growth conditions for several strains of Saccharomyces cerevisiae . The level of variability (standard deviation) was 1-5% between trials where the replications were performed on the same culture using enzyme prepared from the same lot, and 5-15% for different cultures of the same strain . This assay can quantitate differences in cell wall structure (1) between exponentially growing and stationary phase cells, (2) among different S . cerevisiae strains, (3) between S . cerevisiae and Candida albicans, (4) between parental and mutated lines, and (5) between drug- or chemically-treated cells and controls.

J Am Podiatr Med Assoc, 1998 Oct, 88(10), 489 - 92
1998 William J . Stickel Bronze Award . Antifungal activity of Melaleuca alternifolia (tea-tree) oil against various pathogenic organisms; Concha JM et al.; Tea-tree oil (oil of Melaleuca alternifolia) has recently received much attention as a natural remedy for bacterial and fungal infections of the skin and mucosa . As with most naturally occurring agents, claims of effectiveness have been only anecdotal; however, several published studies have recently demonstrated tea-tree oil's antibacterial activity . This study was conducted to determine the activity of tea-tree oil against 58 clinical isolates: Candida albicans (n = 10), Trichophyton rubrum (n = 8), Trichophyton mentagrophytes (n = 9), Trichophyton tonsurans (n = 10), Aspergillus niger (n = 9), Penicillium species (n = 9), Epidermophyton floccosum (n = 2), and Microsporum gypsum (n = 1) . Tea-tree oil showed inhibitory activity against all isolates tested except one strain of E floccosum . These in vitro results suggest that tea-tree oil may be useful in the treatment of yeast and fungal mucosal and skin infections.

Microb Pathog, 1998 Sep, 25(3), 121 - 9
Up-regulation of two Candida albicans genes in the rat model of oral candidiasis detected by differential display; Zhao XJ et al.; Candida albicans is an opportunistic fungal pathogen responsible for the largest percentage of fungal-mediated oral and oesophageal disease . In this regard, knowledge concerning patterns of gene expression during the establishment and/or maintenance of infection may be the key to the design of new strategies for treatment, as well as providing insight into pathogenesis . To address this issue, experiments were performed that utilized differential display to compare the spectrum of C . albicans genes expressed during oral infection versus growth in in vitroculture . Experimentally, the rat model of oral candidiasis served as the in vivo source . After initiation of infection and subsequent harvesting of C . albicans from the rat oral cavity, RNA was isolated, and used with a small number of primers in reverse-transcriptase polymerase chain reaction (RT-PCR) and differential display experiments . Fragments unique to in vivo samples were subcloned and sequenced . Southern blot analysis verified the origin of seven fragments as fromC . albicans . Additionally, specific RT-PCR confirmed that two of these fragments represented genes that were up-regulated during C . albicans in vivo growth in the rat model . Database searches indicated the fragments share homology with a member of the C . albicans agglutinin gene family and to a bacterial gene (gidB) possibly involved in cell division .

J Cardiovasc Surg (Torino), 1998 Aug, 39(4), 437 - 9
A solitary iliac artery aneurysm caused by Candida infection . Report of a case; Tsunezuka Y et al.; We report a very rare case of an infected aneurysm of solitary common iliac artery by Candida albicans . The patient, a 70 year-old male, had a history of systemic Candidemia infected through intravenous hyperlimentation (i.v.H) catheter 2 years ago . By physical examinations and laboratory data, infectious disease was suspected . Computed tomography showed right hydronephrosis and right solitary common iliac artery aneurysm, and operation was performed with diagnosis of infected aneurysm . The aneurysm was removed with the end of the abdominal aorta, and the arterial blood flow was restored by axillo-bifemoral bypass . Histopathological findings revealed abscess formation around the aneurysm with phlogocytes infiltration in both outer media of aneurysmal wall and vasa vasorum . Candida albicans was found as causative pathogen from resected specimens . This aneurysm is considered to be resulted from surviving candida in vasa vasorum after previous candidemia.

Mol Microbiol, 1998 Oct, 30(1), 67 - 81
Molecular cloning and characterization of a Candida albicans gene coding for cytochrome c haem lyase and a cell wall-related protein; Cervera AM et al.; Immunoscreening of a Candida albicans cDNA library with a monoclonal antibody (mAb 4C12) recognizing an epitope present in high-molecular-weight mannoprotein (HMWM) components specific for the mycelial cell walls (a 180 kDa component and a poly-dispersed 260 kDa species) resulted in the isolation of the gene CaCYC3 encoding for cytochrome c haem lyase (CCHL) . The CaCYC3 gene was transcribed preferentially in mycelial cells in which two mRNA transcripts of 0.8 and 1 kb were found . The nucleotide and the deduced amino acid sequences of this gene displayed 45% homology and 46% identity, respectively, to the Saccharomyces cerevisiae CYC3 gene and shared common features with other reported genes encoding for CCHL . The CaCYC3 gene restored the respiratory activity when transformed in a S . cerevisiae cyc3- mutant strain . A C . albicans CYC3 null mutant was constructed after sequential disruption using the hisG::URA3::hisG ('ura-blaster') cassette . Null mutant cells were unable to use lactate as a sole carbon source and had a reduced ability to form germ tubes . Western immunoblotting analysis of subcellular fractions from wild-type and null mutant strains demonstrated the presence of two gene products, a 33kDa mitochondrial protein and a 40 kDa cell wall-associated moiety reacting with antibodies against CCHL, in both yeast cells and germ tubes . mAb 4C12 still reacted with the CaCYC3 null mutant (by immunofluorescence and immunoblotting) but showed an altered pattern of immunoreactivity against cell wall HMWM species, indicating a relationship between these moieties and the CaCYC3 gene products . The results suggest that the CaCYC3 gene encodes two proteins, one targeted to the mitochondria and the other to the cell wall.

Infect Dis Obstet Gynecol, 1998, 6(3), 129 - 33
Prevalence of cervicovaginal infections during gestation and accuracy of clinical diagnosis; Simoes JA et al.; OBJECTIVES: The aim of this study was to establish the prevalence of cervicovaginal infections in normal third-trimester pregnant women and evaluate the accuracy of clinical diagnosis . METHOD: A total of 328 pregnant women were followed at the Prenatal Outpatient Clinic of the Department of Obstetrics and Gynecology at the School of Medical Sciences, Universidade Estadual de Campinas (UNICAMP), Brazil, from October 1991 to February 1993 . The clinical diagnosis was based on the characteristics of the vaginal discharge, and the etiological diagnosis was based on bacterioscopy of the vaginal secretion and direct immunofluorescence for Chlamydia trachomatis . The data were analyzed statistically, determining the sensitivity, specificity, and positive and negative predictive value of the clinical diagnosis related to the laboratory diagnosis of the different infections . RESULTS: The prevalence of infection was 39.6% (Candida albicans, 19.2%; bacterial vaginosis, 9.5%; intermediate vaginal flora, 6.7%; Chlamydia trachomatis, 2.1%; and vaginal trichomoniasis, 2.1%) . The accuracy of clinical diagnosis was low, with sensitivity between 50% and 65% and specificity around 60%, with the exception of trichomoniasis, which showed a sensitivity of 100% and chlamydia, with a sensitivity of 0% and a specificity of 100% . CONCLUSION: The accuracy of the clinical diagnosis of infections was low, specifically with respect to the positive predictive value . The results demonstrate the need for specific testing of cervicovaginal infections at prenatal visits . Reliance on simple vaginal examination results in a low yield for detection of vaginal infections.

Infect Immun, 1998 Nov, 66(11), 5301 - 6
Altered expression of selectable marker URA3 in gene-disrupted Candida albicans strains complicates interpretation of virulence studies; Lay J et al.; The ura-blaster technique for the disruption of Candida albicans genes has been employed in a number of studies to identify possible genes encoding virulence factors of this fungal pathogen . In this study, the URA3-encoded orotidine 5'-monophosphate (OMP) decarboxylase enzyme activities of C . albicans strains with ura-blaster-mediated genetic disruptions were measured . All strains harboring genetic lesions via the ura-blaster construct showed reduced OMP decarboxylase activities compared to that of the wild type when assayed . The activity levels in different gene disruptions varied, suggesting a positional effect on the level of gene expression . Because the URA3 gene of C . albicans has previously been identified as a virulence factor for this microorganism, our results suggest that decreased virulence observed in strains constructed with the ura-blaster cassette cannot accurately be attributed, in all cases, to the targeted genetic disruption . Although revised methods for validating a URA3-disrupted gene as a target for antifungal drug development could be devised, it is clearly desirable to replace URA3 with a different selectable marker that does not influence virulence.

Infect Immun, 1998 Nov, 66(11), 5175 - 82
Immunological characterization of Asp f 2, a major allergen from Aspergillus fumigatus associated with allergic bronchopulmonary aspergillosis; Banerjee B et al.; The 37-kDa recombinant protein Asp f 2, encoding an allergen of Aspergillus fumigatus, was expressed in a prokaryotic expression system and immunologically evaluated for its functional and structural properties . The open reading frame for a 310-amino-acid-long protein was shown to encode a signal peptide of 31 amino acids . A native 37-kDa culture filtrate protein and a 55-kDa mycelial glycoprotein (gp55) exhibited complete N-terminal sequence homology to Asp f 2 . A GenBank search for homologous proteins revealed 60 and 44% sequence homologies to the cytosolic protein ASPND1 from Aspergillus nidulans and fibrinogen binding protein from Candida albicans, respectively . The glycosylation sites and cysteine molecules are conserved in all the three proteins . The extracellular matrix protein laminin showed a dose-dependent interaction with Asp f 2 . This protein, expressed as a major cell-associated protein within 24 h of in vitro fungal culture, comprises 20 to 40% of total fungal protein . Furthermore, both native and recombinant Asp f 2 exhibited specific immunoglobulin (IgE) binding with allergic bronchopulmonary aspergillosis (ABPA) and cystic fibrosis-ABPA patients, whereas A . fumigatus-sensitized allergic asthma and normal control subjects failed to show IgE binding with Asp f 2 . These results indicate that Asp f 2 is a major allergen of A . fumigatus exhibiting IgE antibody binding with sera from patients with ABPA . The antigen should be explored further for its potential role in the differential diagnosis of A . fumigatus-associated allergic diseases.

Biochem Biophys Res Commun, 1998 Sep 29, 250(3), 589 - 92
Ranatuerins: antimicrobial peptides isolated from the skin of the American bullfrog, Rana catesbeiana; Goraya J et al.; Nine peptides, termed ranatuerins 1-9, with antimicrobial activity towards Staphylococcus aureus, were isolated from an extract of the skin of the adult American bullfrog, Rana catesbeiana . In common with other cytolytic peptides from Ranid frogs, (e.g . ranalexin, gaegurins, brevinins), ranatuerins 1 and 4 contain an intramolecular disulfide bridge forming a heptapeptide ring whereas in ranatuerins 2 and 3 the disulfide bridge forms a hexapeptide ring . The structurally related ranatuerins 5-9 comprise 12 - 14 amino acids and show sequence similarity towards the hemolytic peptides A1 and B9 previously isolated from the skin of Rana esculenta . Of the peptides purified, ranatuerin 1 (SMLSVLKNLGKVGLG FVACKINKQC) showed the broadest spectrum of antimicrobial action with inhibitory activity against S . aureus, Escherichia coli and Candida albicans .

J Nat Prod, 1998 Oct, 61(10), 1202 - 8
DNA-damaging steroidal alkaloids from Eclipta alba from the suriname rainforest1; Abdel-Kader MS et al.; Bioassay-guided fractionation of the MeOH extract of Eclipta alba using three yeast strains (1138, 1140, and 1353) resulted in the isolation of eight bioactive steroidal alkaloids (1-8), six of which are reported for the first time from nature . The major alkaloid was identified as (20S)(25S)-22,26-imino-cholesta-5,22(N)-dien-3beta-ol (verazine, 3), while the new alkaloids were identified as 20-epi-3-dehydroxy-3-oxo-5,6-dihydro-4,5-dehydroverazine (1), ecliptalbine {(20R)-20-pyridyl-cholesta-5-ene-3beta,23-diol} (4), (20R)-4beta-hydroxyverazine (5), 4beta-hydroxyverazine (6), (20R)-25beta-hydroxyverazine (7), and 25beta-hydroxyverazine (8) . Ecliptalbine (4), in which the 22,26-imino ring of verazine was replaced by a 3-hydroxypyridine moiety, had comparable bioactivity to verazine in these assays, while a second alkaloid (8) showed good activity against Candida albicans . All the alkaloids showed weak cytotoxicity against the M-109 cell line.

Microbiology, 1998 Sep, 144 ( Pt 9), 2417 - 26
Cloning and sequencing of the Candida albicans homologue of SRB1/PSA1/VIG9, the essential gene encoding GDP-mannose pyrophosphorylase in Saccharomyces cerevisiae; Warit S et al.; Two genomic fragments have been isolated from Candida albicans which strongly hybridize to SRB1/PSA1/VIG9, an essential gene which encodes GDP-mannose pyrophosphorylase in Saccharomyces cerevisiae . A common 2.5 kb Xbal-Pstl fragment has been identified, which Southern analysis suggests is most likely unique in the C . albicans genome . The fragment contains an ORF, which is 82% identical and 90% homologous to the Srb1p/Psa1p/Vig9p from S . cerevisiae, contains one additional amino acid at position 254 and is able to functionally complement the major phenotypic characteristics of S . cerevisiae srb1 null and conditional mutations . The authors therefore conclude that they have cloned and sequenced from C . albicans the bona fide homologue of SRB1/PSA1/VIG9, named hereafter CaSRB1 . Northern analysis data indicate that the gene is expressed in C . albicans under conditions of growth in the yeast and hyphal form and suggest that its expression might be regulated.

Adv Exp Med Biol, 1998, 443, 229 - 37
Enhanced anti-Candida activity of neutrophils and azole antifungal agents in the presence of lactoferrin-related compounds; Wakabayashi H et al.; We investigated the effects of lactoferrin (Lf)-related compounds on growth inhibition of Candida albicans by neutrophils or antifungal agents in vitro . Human neutrophils partially inhibited the growth of C.albicans . The growth inhibition caused by human neutrophils was augmented by the addition of human Lf at concentrations which did not show any inhibitory effect in the absence of neutrophils . Similar observations were obtained also with the following combinations: human neutrophils + bovine Lf, murine neutrophils + bovine Lf, and murine neutrophils + iron saturated bovine Lf, but not in the case of murine neutrophils + human transferrin . The minimum inhibitory concentration (MIC) of azole antifungal agents, clotrimazole, ketoconazole, fluconazole, and itraconazole was reduced by 1/4 to 1/16 in the presence of a sub-MIC level of each of bovine Lf, bovine Lf pepsin hydrolysate, and the antimicrobial peptide "lactoferricin B" (Lfcin B) . Other types of antifungal agents, amphotericin B, nystatin, and flucytosine did not show such combined effects with these Lf-related compounds . The anti-Candida activity of bovine Lf or Lfcin B in combination with clotrimazole was shown to be synergistic by checkerboard analysis . Clinically isolated azole-resistant C . albicans strains were more susceptible to bovine Lf or Lfcin B than azole-susceptible strains . Trailing growth of an azole-resistant strain in the presence of fluconazole was reduced by the addition of sub-MIC levels of bovine Lf or Lfcin B . These results suggest that Lf-related compounds even at relatively low concentrations may function as an antifungal effector in combination with neutrophils thereby modulating azole antifungal efficacies in vivo.

Kansenshogaku Zasshi, 1998 Aug, 72(8), 813 - 9
{Antifungal susceptibility of clinically isolated Candida albicans by broth microdilution method}; Yamazumi T et al.; In this study we investigated the antifungal susceptibility of 285 strains of Candida albicans isolates at Kinki University Hospital from March 1995 to December 1996 . The antifungal agents tested were fluconazole, miconazole, intraconazole, amphotericin B and flucytosine . The susceptibility testing were performed according to the broth microdilution method standardized by National Committee for Clinical Laboratory Standards (M27-T) . Most isolates of C . albicans showed relatively a low MIC value and the MIC90S were calculated at 1 microgram/ml; fluconazole, 0.125 microgram/mg; miconazole, 0.06 microgram/ml; itraconazole, 1 microgram/ml; amphotericin B, 0.25 microgram/ml; flucytosine . There was only one strain that showed high resistance against fluconazole and it showed cross-resistance against miconazole and itraconazole . There were two flucytosine resistant strains . The MICs of amphotericin B were tightly clustered and resistant strain were not observed.

Med Mycol, 1998 Aug, 36(4), 213 - 7
Epidemiology of Candida albicans isolates from heroin addicts analysed by DNA typing; McCullough MJ et al.; Candida albicans is a ubiquitous commensal organism of humans . Several studies have examined outbreaks of candidiasis in heroin addicts utilizing a variety of methods to assess the epidemiological relatedness of the isolates and suggested the association of certain subtypes with disease in this patient population . The aim of the present study was to assess a separate group of isolates of C . albicans from heroin addicts in Spain using a DNA typing method . Results showed that, of the 34 isolates from heroin addicts, 20 were in subgroup IA, 10 were in subgroup IB and no isolates were of the subtype IA2 . In addition, four isolates were in a recently described subgroup IC . Control isolates from the same geographical region (Spain) showed a distribution similar to the Spanish heroin addict isolates (12 subgroup IA, three subgroup IB, two subgroup IC and no isolates of the subtype IA2) . In this study isolates from the same locality appeared similar irrespective of the patient population from which they were isolated . These results indicated that there may be differing geographical diversity of C . albicans than has previously been reported and that the newly described genotypic subgroup (IC) of C . albicans may be more widespread than previously shown.

Med Mycol, 1998 Apr, 36(2), 123 - 5
Promotion of chlamydoconidium formation in Candida albicans by corn meal broth incubation; Nakamoto S; Chlamydoconidium formation can be used as a tool for the identification of Candida albicans . While chlamydoconidia are known to be inducible on corn meal agar, this report demonstrates that testing in liquid media supplemented with milk or serum enhances chlamydoconidium formation and the formation of complex mycelial clusters.

Med Mycol, 1998 Apr, 36(2), 81 - 7
UV irradiation induced high frequency of colonial variants with altered morphology in Sporothrix schenckii; Torres-Guerrer H et al.; Ultraviolet light (UV) exposure of Sporothrix schenckii strains resulted in a high frequency of morphological variants that ranged from 10(-3) to 10(-1) depending on the strain and dose of UV . Based on their morphological differences, these variants were classified into five different groups . One common feature among them was that they were smaller in size compared to the wild type . Two morphological phenotypes (II and IV) were fuzzy, like the wild-type colony, and only the colony size was altered . Phenotypes I, III and V had different shapes; they lost the fuzzy appearance and the individual hyphae in the colony were of aberrant shape . Stable and non-stable morphological variants were found in the population; reversion of the mutant phenotype was always to the wild-type phenotype . Unlike Candida albicans, phenotypic switching was not found in individual colonial phenotypes.

Med Mycol, 1998 Feb, 36(1), 29 - 36
Analysis of fluconazole effect on Candida albicans viability during extended incubations; Sohnle PG et al.; Fluconazole is an azole agent with primarily fungistatic activity in standard in vitro susceptibility tests . However, recent work has demonstrated that this drug can reduce Candida albicans viability during prolonged incubations under non-growing conditions . The present study was undertaken to examine more closely some of the parameters of this killing activity . Fungicidal effects of 1.0 microg ml-1 of fluconazole were found during 7-14-day exposures in each of two media that prevented proliferation, distilled water and metal-depleted RPMI 1640 tissue-culture medium . Fluconazole appeared to be stable after being incubated at 37 degreesC for either 7 or 14 days . Strains of C . albicans resistant to fluconazole in standard short-term growth-inhibition assays were also found to be resistant to fluconazole's effect on viability in prolonged culture, suggesting similar mechanisms of action for these effects . C . albicans yeast cells pre-incubated for 7 days in distilled water were not more sensitive to the drug in short-term susceptibility assays . Although all proliferation of the organisms in distilled water cultures appeared to cease after 3 days, fluconazole added at 7 days still reduced C . albicans viability . Therefore, the drug appeared to kill the non-proliferating organisms directly rather than preventing growth and thereby the emergence of younger organisms that would live longer . Transmission electron microscopy demonstrated damage to the cell wall-cell membrane complex and interior contents of yeast cells incubated in distilled water alone; fluconazole appeared to increase the percentages of cells so affected . In summary, extended-incubation susceptibility tests demonstrated that fluconazole has direct fungicidal activity of non-proliferating C . albicans yeast cells . These results may be relevant to the manner in which this drug promotes clearance of chronic fungal infections.

J Clin Microbiol, 1998 Nov, 36(11), 3260 - 5
Rapid identification of Candida species with species-specific DNA probes; Elie CM et al.; Rapid identification of Candida species has become more important because of an increase in infections caused by species other than Candida albicans, including species innately resistant to azole antifungal drugs . We previously developed a PCR assay with an enzyme immunoassay (EIA) format to detect amplicons from the five most common Candida species by using universal fungal primers and species-specific probes directed to the ITS2 region of the gene for rRNA . We designed probes to detect seven additional Candida species (C . guilliermondii, C . kefyr, C . lambica, C . lusitaniae, C . pelliculosa, C . rugosa, and C . zeylanoides) included in the API 20C sugar assimilation panel, five probes for species not identified by API 20C (C . haemulonii, C . norvegica, C . norvegensis, C . utilis, and C . viswanathii), and a probe for the newly described species C . dubliniensis, creating a panel of 18 Candida species probes . The PCR-EIA correctly identified multiple strains of each species tested, including five identified as C . albicans by the currently available API 20C database but determined to be C . dubliniensis by genotypic and nonroutine phenotypic characteristics . Species identification time was reduced from a mean of 3.5 days by conventional identification methods to 7 h by the PCR-EIA . This method is simple, rapid, and feasible for identifying Candida species in clinical laboratories that utilize molecular identification techniques and provides a novel method to differentiate the new species, C . dubliniensis, from C . albicans.

Ann Cardiol Angeiol (Paris), 1998 Jul-Sep, 47(7), 465 - 8
{Candida albicans myocarditis or endocarditis? Echocardiographic aspects}; Donal E et al.; The authors report the case of a patient presenting with atypical features of Candida albicans cardiac infection, with unusual infectious destructive lesions . While the initial hypothesis was that of aortic endocarditis, the clinical course and operative findings showed echocardiographic features of septal micro-abscesses adjacent to the aortic orifice, which was devoid of any infectious lesions.

Pathol Biol (Paris), 1998 Jan, 46(1), 34 - 8
{Validation of a sampling method for the channels of a flexible endoscope which is experimentally contaminated}; Luu Duc D et al.; The increase of endoscopic actions and infectious risks explains of interest in endoscopic material maintenance processing . This purpose study was to set to the point and validate a microbiological flexibles endoscopes channels (suction, biopsy, air/water) sample method . An inoculum made of faeces contents material and microorganisms suspension (Pseudomonas aeruginosa or Candida albicans), at low or high concentration, was injected into digestive endoscope's channels . For each microorganism, five trials were carried out at both of the two concentrations . After incubation, a global channels sample was performed with a recovering solution, then a counting was made . A variance analysis (ANOVA) concerning restated measures was performed for the whole results . The mean difference between the number of microorganisms injected and those recovered's is 0.114 log (IC95% {+0.088; +0.140}) . It does not vary significantly according to the micro-organisms (p = 0.69) and the inoculum (p = 0.70) . The coefficient of variation which is 0.525 (min: 0.010; max: 0.260) shows this method can be reproduced . This technique are allowed to be recommended to evaluate and to validate manuals and automatics endoscopes maintenance processing.

Pathol Biol (Paris), 1998 May, 46(5), 325 - 9
{Efficacy of an ultraviolet device for the disinfection of radiology cassettes}; Garcin F et al.; The antimicrobial efficiency of a disinfection box of radiology cassettes, by means of a device which emits ultraviolet rays, was studied according to a method defined by the laboratory . Germ-carriers contaminated with Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans were irradiated and compared to non irradiated reference ones . The bactericidal and fungicidal efficiency was better than 99.9% in 5 minutes of irradiation . This procedure represents an interesting alternative to the chemical treatment of radiology cassettes.

Pathol Biol (Paris), 1998 May, 46(5), 307 - 14
Oropharyngeal candidiasis in AIDS patients from Abidjan (Ivory Coast): antifungal susceptibilities and multilocus enzyme electrophoresis analysis of Candida albicans isolates; Nebavi F et al.; Multilocus enzyme electrophoresis (MEE) and in vitro antifungal susceptibility testing were used to investigate the Candida albicans strain diversity in twenty nine AIDS patients from Abidjan (Ivory Coast) . All patients were monitored for a first episode of oropharyngeal candidiasis and were randomly clustered into three groups of therapy: ketoconazole, amphotericin B or nystatin . Oral swabs were collected before every treatment, 14 and 30 days after the initiation of the therapy; a total of 67 isolates were investigated . No resistant or less susceptible isolate to any antifungal agent was found despite the emergence of clinical relapses, mainly for patients treated with nystatin or amphotericin B . The MEE analysis revealed 27 different electrophoretic types (ETs) . Genetic distances between ETs were statistically analyzed and represented on a dendrogram . The 27 ETs clustered into three groups; in each group, ETs represented variants of the same strain . A segregation of the C . albicans isolates seemed to be as a function of the serotype.

Curr Microbiol, 1998 Nov, 37(5), 359 - 61
Isolation of adenylate cyclase gene-specific sequences from Ophiostoma novo-ulmi, Candida albicans, and Agaricus bisporus by PCR; Binz T et al.; Degenerate primers corresponding to consensus sequences in the catalytic domains of known fungal adenylate cyclases were used to isolate gene-specific homologs from the Dutch elm disease pathogen Ophiostoma novo-ulmi, the dimorphic human pathogen Candida albicans, and the commercial mushroom Agaricus bisporus . All three fungi gave the expected PCR product of about 390 bp . Computer searches of the databases revealed that the products generated from O . novo-ulmi and C . albicans were highly similar to the adenylate cyclase gene of Magnaporthe grisea, the rice blast fungus (91% and 79%, respectively) . The PCR product from the homobasidiomycete A . bisporus, on the other hand, showed 78% similarity to the uac1 gene of the heterobasidiomycete smut fungus, Ustilago maydis . Southern hybridization indicated that all three fungi contain a single adenylate cyclase gene . Our data suggest that PCR will be highly successful for the isolation of adenylate cyclase sequences from other fungi.

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