J Clin Microbiol, 2000 Jun, 38(6), 2334 - 8 Comparison of a new colorimetric assay with the NCCLS broth microdilution method (M-27A) for antifungal drug MIC determination; Li RK et al.; We evaluated a new microtiter assay for antifungal susceptibility testing based on a colorimetric reaction to monitor fungal substrate utilization . This new method (rapid susceptibility assay {RSA}) provides quantitative endpoint readings in less than 8 h compared with visual determination of MIC by the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution method, which requires a minimum of 48 h of incubation . In this study, we tested clinical isolates from each of the following species: Candida albicans (20 isolates), C . glabrata (20 isolates), C . krusei (19 isolates), C . tropicalis (19 isolates), and C . parapsilosis (28 isolates) . RSA and NCCLS broth dilution methods were used to determine the MICs of amphotericin B, fluconazole, itraconazole, and 5-flucytosine for all 106 isolates . RPMI 1640 medium buffered with morpholinopropanesulfonic acid was used for both methods; however, glucose and inoculum concentrations in the RSA were modified . RSA MICs were determined as the lowest drug concentration that prevented glucose consumption by the organism after 6 h of incubation . MICs obtained from the RSA were compared with those obtained from the NCCLS M-27A method read at 24 and 48 h . MIC pairs were considered in agreement when the difference between the pairs was within 2 twofold dilutions . For the 106 isolates tested, amphotericin B and 5-flucytosine demonstrated the highest agreement in MICs between the two methods (100 and 98%, respectively), whereas fluconazole and itraconazole produced less favorable MIC agreement (63.2 and 61.3%, respectively) . The azole MIC differences between the two methods were significantly reduced when lower inocula were used with a prolonged incubation time . This preliminary comparison suggests that this rapid procedure may be a reliable tool for the in vitro determination of MICs of amphotericin B and 5-flucytosine and warrants further evaluation. J Clin Microbiol, 2000 Jun, 38(6), 2317 - 23 Intestinal lesions associated with disseminated candidiasis in an experimental animal model; Andrutis KA et al.; In human patients, disseminated candidiasis, a life-threatening disease for immunocompromised patients, is often associated with intestinal lesions . In this study, we demonstrate that immunosuppressed gnotobiotic (IGB) piglets orally inoculated with wild-type Candida albicans developed extensive intestinal lesions and disseminated infection . Severe ulceration of the ileal mucosa was observed overlying regions of colonization and necrosis of the gut-associated lymphoid tissue . Despite the high susceptibility of IGB piglets to many microbial pathogens, an avirulent mutant strain of C . albicans failed to produce intestinal lesions and exhibited poor dissemination, demonstrating that these effects required virulent organisms . It is likely that in IGB piglets, as in human patients, intestinal lesions provide the mechanism for escape of C . albicans from the gastrointestinal tract . Multinucleated giant cells containing fungal organisms were observed within lymph nodes and lymphatic vessels, and as with other pathogens, such cells could provide a mechanism for dissemination of C . albicans. Cornea, 2000 May, 19(3), 307 - 12 Spectrum of fungal keratitis at Wills Eye Hospital, Philadelphia, Pennsylvania; Tanure MA et al.; PURPOSE: To report the spectrum of fungal keratitis at Wills Eye Hospital, Philadelphia . METHODS: We reviewed the records of 24 cases of culture-positive fungal keratitis treated from January 1991 to March 1999 at Wills Eye Hospital . Risk factors, fungal identification, antifungal treatment, and outcomes were evaluated . RESULTS: The study included 24 eyes (24 patients) . Fourteen patients (58.3%) were female . The mean age was 59 years (range, 19-86 years) . Predisposing factors included chronic ocular surface disease (41.7%), contact lens wear (29.2%), atopic disease (16.7%), topical steroid use (16.7%), and ocular trauma (8.3%) . Early identification of fungal elements was achieved by staining of corneal scrapings in 18 cases (75%) . Half of the cases (12 eyes) had corneal infections caused by yeast, and the other half by filamentous fungi . Candida albicans was the most commonly isolated organism (45.8%), followed by Fusarium sp (25%) . Natamycin and amphotericin B were the topical antifungals most frequently used, while systemic treatment commonly used included fluconazole, ketoconazole, or itraconazole . Six patients (25%) had penetrating keratoplasty during the acute stage of infection . After a mean follow-up of nine months, 13 eyes (54.1%) had the best corrected visual acuity 20/100 or better . CONCLUSIONS: In contrast to other studies from the northern United States, we found Fusarium sp the most commonly isolated filamentous fungus . In our series, C . albicans was the most frequent cause of fungal keratitis, and a past history of ocular trauma was uncommon. Microbiology, 2000 May, 146 ( Pt 5), 1035 - 44 The Candida albicans antiporter gene CNH1 has a role in Na+ and H+ transport, salt tolerance, and morphogenesis; Soong TW et al.; The isolation and functional characterization of a Candida albicans Na+/H+ antiporter gene, CNH1, is reported here . The gene encodes a protein of 840 amino acids that exhibits high levels of similarity in sequence, size, and structural and functional domains to a group of known Na+/H+ antiporters of fungi . The CNH1 gene is able to functionally complement the salt-sensitivity of a Saccharomyces cerevisiae ena1 nha1 mutant, and mutations of two conserved aspartate residues to asparagines in the putative Na+-binding site abolish this activity . Deletion of CNH1 results in retardation of growth and a highly elongated morphology in a significant fraction of cells under conditions that normally support yeast growth . These results indicate that CNH1 has a role in Na+ and H+ transport, salt-tolerance, and morphogenesis. Biometals, 2000 Mar, 13(1), 1 - 8 Synthesis and antimicrobial activity of copper(II) and manganese(II) alpha,omega-dicarboxylate complexes; Geraghty M et al.; Copper(II) alpha,omega-dicarboxylate complexes of general formulae, {Cu(O2C(CH2)nCO2)}.xH2O, {Cu(O2C(CH2)nCO2) (phen)2}.xH2O and {Cu(O2C(CH2),CO2)(bipy)y}.xH2O (n = 1-8; y = 1, 2; phen = 1,10-phenanthroline; bipy = 2,2'-bipyridine) were synthesised . These copper complexes, some related manganese(II) complexes and the metal-free ligands were screened in vitro for their ability to inhibit the growth of Candida albicans . Metal-free 1,10-phenanthroline and all of the copper(II) and manganese(II) phenanthroline complexes were potent growth inhibitors, with only one bipyridine complex, {Cu(O2C(CH2)CO2)(bipy)2}.2H2O, having moderate activity . The remaining substances were effectively inactive . Complexes which were active against C . albicans also proved effective against C . glabratta, C . tropicalis and C . kreusi with the manganese complexes retaining superior activity . For the phenanthroline complexes the active drug species is thought to be the dication {M(phen)2(H2O)n}2+ (M = Cu, Mn) . Escherichia coli and Staphylococcus aureus were resistant to all of the metal complexes and also to metal-free 1,10-phenanthroline . Only the copper phenanthroline complexes showed intermediate activity against Pseudomonas aeruginosa. J Asthma, 2000 May, 37(3), 281 - 90 Inhibitory activity of fenoterol on Dermatophagoides-, Parietaria-, tetanus-toxoid-, and Candida albicans-stimulated blood mononuclear cells: differences in beta2-adrenoreceptor stimulation but not in cell apoptosis; Silvestri M et al.; beta2-adrenoreceptor agonists have the ability to downregulate in vitro the proliferative response of peripheral blood mononuclear cells (BMCs) . This activity could be related to a variety of beta2-adrenoreceptor-mediated functions, including induction of cell apoptosis in activated T-cells . To test this hypothesis, BMCs from atopic subjects, sensitized to house dust mites (Dermatophagoides {Der p}) and/or to Parietaria were incubated with fenoterol (10(-8)-10(-5) M) in the presence of (a) purified allergen extracts (Der p {5 microg/mL} or Parietaria {5 microg/mL}) or (b) antigens (tetanus toxoid {1 microg/mL} or Candida albicans {5 x 10(5) bodies/mL}) . The BMC proliferation was assessed by {3H} thymidine incorporation and cell apoptosis was assessed by evaluating DNA fragmentation by a fluorescence technique, using propidium iodide . In cultures stimulated with Der p or with Parietaria, fenoterol induced a dose-dependent inhibition of BMC proliferation, significant also at the lowest concentration tested (10(-8) M) (p < 0.05, each comparison) . In contrast, the inhibitory activity of the drug on tetanus-toxoid-stimulated BMCs was significant only at the highest dose tested (10(-5)M) (p < 0.05), whereas no effect was seen when BMCs were stimulated with C . albicans extract (p > 0.05) . The different inhibitory efficacy of fenoterol appeared to be related to the degree of activation of beta2-adrenoreceptors on the different BMC populations that responded to the different stimuli . Indeed, in the presence of fenoterol (10(-6) and 10(-5)M), a significant increase in cyclic adenosine monophosphate (cAMP) levels was seen in Der p- or Parietaria-stimulated cells (p < 0.05; each comparison), but not in cell cultures stimulated with tetanus toxoid or with C . albicans extracts (p > 0.05; each comparison) . Finally, the percentage of cells with fragmented DNA was lower in cultures stimulated with Der p or Parietaria than in those stimulated with tetanus toxoid or C . albicans, and the presence of fenoterol did not modify cell apoptosis (p > 0.05; each comparison) . Thus, the different inhibitory activity of fenoterol on BMCs activated by allergens (Der p or Parietaria) or by antigens (tetanus toxoid or C . albicans) seems to be related to differences in beta2-adrenoreceptor expression and/or function in the different antigen-specific T-cell subsets, but it is not influenced by changes in cell apoptosis. Regul Pept, 2000 Jun 30, 90(1-3), 53 - 60 Purification and characterization of antimicrobial and vasorelaxant peptides from skin extracts and skin secretions of the North American pig frog Rana grylio; Kim JB et al.; Eight peptides with differential growth-inhibitory activity against the gram-positive bacterium Staphylococcus aureus, the gram-negative bacterium Escherichia coli and the yeast, Candida albicans were isolated from an extract of the skin of the North American pig frog Rana grylio . The primary structures of these antimicrobial peptides were different from previously characterized antimicrobial peptides from Ranid frogs but on the basis of sequence similarities, the peptides may be classified as belonged to four previously characterized peptide families: the ranatuerin-1, ranatuerin-2 and ranalexin families, first identified in the North American bullfrog, Rana catesbeiana, and the temporin family first identified in the European common frog Rana temporaria . Peptides belonging to the brevinin-1, brevinin-2, esculentin-1, and esculentin-2 families, previously isolated from the skins of other species of Ranid frogs, were not identified in the extracts . The ranatuerin-1 and ranalexin peptides showed broadest spectrum of antimicrobial activity whereas the temporins were active only against S . aureus . Synthetic replicates of temporin-1Gb (SILPTIVSFLSKFL.NH(2)) and temporin-1Gd (FILPLIASFLSKFL.NH(2)) produced concentration-dependent relaxation of preconstricted vascular rings from the rat thoracic aorta (EC(50) = 2.4+/-0.1 microM for temporin-1Gb and 2.3+/-0.2 microM for temporin-1Gd) . The antimicrobial peptides that were isolated in extracts of the skin R . grylio were present in the same molecular forms in electrically-stimulated skin secretions of the animal demonstrating that the peptides are stored in the granular glands of the skin in their fully processed forms. Gerodontology, 1999 Dec, 16(2), 110 - 8 Efficacy of antifungal agents in tissue conditioners in treating candidiasis; Chow CK et al.; Chronic atrophic candidiasis is prevalent in up to 72% of institutionalized geriatric populations and is causally associated with Candida albicans . Topical antifungal treatments are difficult to implement in some geriatric patients due to cognitive impairment, reduced motor dexterity and memory loss . OBJECTIVE: This in vitro study incorporated antifungal agents into tissue conditioners to investigate the effectiveness of this method of drug delivery . DESIGN: Combinations of nystatin, fluconazole, itraconazole and Coe Soft, Viscogel, Fitt were tested at 1, 3, 5, 7, 9 and 11 wt/wt%, with and without sterilized saliva . 6 mm diameter cores were punched in Sabouraud plates pre-grown with standardized C . albicans . Antifungal agents plus tissue conditioner mixtures were injected into each core . Inhibition diameters were measured for 14 days . RESULTS: Cores with only tissue conditioners acted as negative control and showed no significant inhibition activity (ANOVA, p > 0.05) . Peak activity was between 65 to 89 hours; followed by a plateau . Itraconazole had greater fungicidal activity than fluconazole; while nystatin was found to have the least fungicidal activity (ANOVA, p < 0.05) . The most effective concentration for nearly all combinations was 5% wt/wt (ANOVA, p < 0.05) . Specimens with saliva showed greater antifungal activity than those without (t-test, p < 0.001) . Itraconazole altered the physical properties of Viscogel hence this combination is not recommended for clinical use . CONCLUSION: The treatment of chronic atrophic candidiasis by incorporation of antifungal drugs into tissue conditioners is efficacious . 5% wt/wt itraconazole mixed with Coe Soft or Fitt is recommended for clinical study where compliance of patient or care giver cannot be relied upon . Peak antifungal activity at 3 days suggests that mixtures prepared for clinical study may be replaced soon after this time for maximum effectiveness. Comp Biochem Physiol B Biochem Mol Biol, 2000 May, 126(1), 109 - 20 Enolase from Candida albicans--purification and characterization; Kustrzeba-Wojcicka I et al.; This paper describes isolation of electrophoretically homogenous enolase from Candida albicans . The purification involved: disintegration of C . albicans cells in a Braun's mill (67-100%) ammonium sulfate precipitation, chromatography on DEAE-Sephadex A-50 at pH 9.0 and chromatography on CM-Sephadex A-50 at pH 6.2 . The procedure resulted in a 30-fold purification of the enzyme with a recovery rate of 6% and a specific activity 35 U mg-1 . The subunit molecular weight was 46 kDa and the pH optimum of the enzyme was 6.8 . Km and Vmax values for the 2PGA-->PEP reaction were determined (Km = 0.95 mM, Vmax = 4200 mumol min-1 mumol-1) . In the absence of orthophosphate, inhibition by fluoride was competitive, which became noncompetitive in the presence of phosphate . It was confirmed that Mg2+ is the most potent activator (Km = 0.286 mM); Mn2+ gave less activity and Zn2+ less still . It was also demonstrated that the presence of two types of cations in the reaction mixture nullified the activatory effect of the stronger agent . Properties of the enzyme from C . albicans are compared with those of enolases from other sources. J Biol Chem, 2000 Aug 4, 275(31), 23725 - 8 Site-directed mutagenesis of the yeast resolving enzyme Cce1 reveals catalytic residues and relationship with the intron-splicing factor Mrs1; Wardleworth BN et al.; The Holliday junction-resolving enzyme Cce1 is a magnesium-dependent endonuclease, responsible for the resolution of recombining mitochondrial DNA molecules in Saccharomyces cerevisiae . We have identified a homologue of Cce1 from Candida albicans and used a multiple sequence alignment to predict residues important for junction binding and catalysis . Twelve site-directed mutants have been constructed, expressed, purified, and characterized . Using this approach, we have identified basic residues with putative roles in both DNA recognition and catalysis of strand scission and acidic residues that have a purely catalytic role . We have shown directly by isothermal titration calorimetry that a group of acidic residues vital for catalytic activity in Cce1 act as ligands for the catalytic magnesium ions . Sequence similarities between the Cce1 proteins and the group I intron splicing factor Mrs1 suggest the latter may also possess a binding site for magnesium, with a putative role in stabilization of RNA tertiary structure or catalysis of the splicing reaction. Hum Immunol, 2000 Jun, 61(6), 531 - 7 Cd4(+) T cell response to Leishmania spp . in non-infected individuals; Gabaglia CR et al.; T cell mediated immunity is known to play a central role in the host response to control intra-cellular pathogens . This work demonstrates the presence of specific CD4(+) T cells to Leishmania spp . antigens in peripheral mononuclear cells of naive individuals (normal volunteers from non-endemic regions) . The responder population was expanded by generation of antigen-specific T cell lines, which were produced by repeated stimulation with fixed promastigotes and autologous irradiated PBMC as antigen presenting cells . The leishmania-T cell lines were shown to proliferate in response to different species of the parasite (L . amazonensis, L . braziliensis, and L . donovani), but not to other recall antigens such as Candida albicans or tetanus toxoid . A preferential expansion of IFNgamma and IL-2 producing Th1-like T cells was observed . The leishmania-reactive cells were distributed between CD4(+) CD45RA(+) ("naive") and CD4(+) CD45R0(+) ("memory") populations . Although limiting dilution analysis showed a precursor frequency 3 times lower within the naive compartment, similar numbers of T cell lines were derived from both purified subpopulations . This study using leishmania-specific CD4(+) T cell lines produced from normal individuals should provide information on cellular immune responses that are triggered by the parasite and how infection impacts the naive T cell repertoire. Oral Dis, 2000 May, 6(3), 166 - 71 The effect of brief exposure to sub-therapeutic concentrations of chlorhexidine gluconate on the germ tube formation of oral Candida albicans and its relationship to post-antifungal effect; Ellepola AN et al.; OBJECTIVES: Adherence of Candida albicans has been implicated as the first step in the pathogenesis of oral candidosis, and germ tube formation a contributory attribute . Recently, these organisms have also been implicated in persistent apical periodontitis . Chlorhexidine gluconate is by far the commonest antiseptic mouth wash prescribe in dentistry . As the intraoral concentrations of this antiseptic fluctuate considerably due to the dynamics of the oral cavity, the main objective of this study was to investigate the effect of brief exposure to three different sub-therapeutic concentrations of chlorhexidine gluconate (0.005%, 0.0025% and 0.00125%) on the germ tube formation of C . albicans . These findings were then correlated with the chlorhexidine-induced post-antifungal effect (PAFE) values we obtained in a study using the identical organisms and experimental conditions . DESIGN: Ten oral isolates of C . albicans were exposed to three different concentrations of chlorhexidine gluconate for 30 min, the antiseptic removed, and the germ tube formation of these isolates quantified following subsequent incubation in a germ tube inducing medium . The PAFE was evaluated by turbidometric measurement of growth . RESULTS: When compared with the controls, exposure to 0.005%, 0.0025% and 0.00125% chlorhexidine gluconate suppressed the ability to form germ tubes by 81.23% (P < 0.01), 42.74% (P < 0.01) and 9.13% (P > 0.05), respectively, while eliciting a mean PAFE of 9.91 h, 1.65 h and 0.67 h respectively . On regression analysis a significant positive correlation was observed between these two parameters (P < 0.0001; r = 0.7325) . CONCLUSIONS: Taken together, these findings imply that short exposure to sub-therapeutic levels of chlorhexidine gluconate may modulate candidal germ tube formation as well as its growth, thereby suppressing its pathogencity in vivo. Peptides, 2000 Apr, 21(4), 469 - 76 Purification and characterization of antimicrobial peptides from the skin of the North American green frog Rana clamitans; Halverson T et al.; Ten peptides with differential growth-inhibitory activity against the gram-positive bacterium, Staphylococcus aureus, the gram-negative bacterium, Escherichia coli, and the yeast Candida albicans were isolated from an extract of the skin of a North American frog, the green frog Rana clamitans . Ranatuerin-1C (SMLSVLKNLGKVGLGLVACKINKQC), ranalexin-1Ca (FLGGLMKAFPALICAVTKKC), ranalexin-1Cb (FLGGLMKAFPAIICAVTKKC), ranatuerin-2Ca (GLFLDTLKGAAKDVAGKLLEGLKCKIAGC KP), and ranatuerin-2Cb (GLFLDTLKGLAGKLLQGLKCIKAGCKP), are members of three previously characterized families of antimicrobial peptides, first identified in the North American bullfrog Rana catesbeiana . In addition, five structurally related peptides (temporin-1Ca, -1Cb, -1Cc, -1Cd, and -1Ce), comprising 13 amino acid residues and containing a C-terminally alpha-amidated residue, belong to the temporin family first identified in the European common frog Rana temporaria . Peptides belonging to the brevinin-1, brevinin-2, esculentin-1, and esculentin-2 families, previously isolated from the skins of Asian and European Ranid frogs, were not identified in the extract . The data support the hypothesis that the distribution and amino acid sequences of the skin antimicrobial peptides are valuable tools in the identification and classification of Ranid frogs. Biol Pharm Bull, 2000 May, 23(5), 672 - 6 Solubilized cell wall beta-glucan, CSBG, is an epitope of Candida immune mice; Uchiyama M et al.; Antibody to beta-glucan is generally difficult to produce in mice . We have recently developed a protocol to obtain a soluble Candida spp . beta-(1-->3)-D-glucan (CSBG) by sodium hypochlorite (NaClO) oxidation and subsequent dimethyl sulfoxide (Me2SO) extraction . CSBG is composed mainly of beta-(1-->3) and beta-(1-->6)-glucosidic linkages with a small amount of branch . In this paper, mice were immunized with Candida albicans and the specificity of the resulting sera to CSBG was examined by ELISA . Using CSBG coated plate, sera of the Candida immune mice showed higher reactivity than non-immune, normal mice and the reactivity was neutralized by adding soluble CSBG as a competitor . However, the reactivity could not be neutralized by a beta-(1-->6) branched beta-(1-->3)-glucan, grifolan . Similar specificity of the sera was obtained by commercially available beta-glucan particle, zymosan or zymocel, immune mice . These facts strongly suggested that CSBG included epitopes of the specific antibody in Candida immune mice. J Agric Food Chem, 2000 May, 48(5), 1875 - 81 Structure elucidation and antifungal activity of an anthracycline antibiotic, daunomycin, isolated from Actinomadura roseola; Kim BS et al.; The actinomycete strain Ao108 producing antifungal metabolites active against some plant pathogenic fungi was identified as Actinomadura roseola, based on the analyses of morphological and physiological characteristics . The antibiotic Da2B that showed a strong antifungal activity was isolated from the culture broth and mycelial mats of A . roseola strain Ao108 using various chromatographic procedures . On the basis of (1)H NMR, (13)C NMR, and 2-D NMR correlation data, the antibiotic Da2B was confirmed to have the structure of an anthracycline antibiotic, daunomycin . In vitro antimicrobial spectrum tests showed that the antibiotic Da2B had substantial inhibitory activity (10 microg mL(-)(1) of MICs) against mycelial growth of Phytophthora capsici and Rhizoctonia solani . The antibiotic also showed antiyeast activity against Saccharomyces cerevisiae, but the growth of Candida albicans was not affected . Antibacterial activity was found only against Gram-positive bacteria . In the further evaluation of in vivo efficacy, application of the antibiotic Da2B effectively inhibited the development of Phytophthora blight in pepper plants . However, the control efficacy of the antibiotic against Phytophthora infection was somewhat less than that of metalaxyl . The antibiotic Da2B did not show any phytotoxicity on pepper plants even at 500 microg mL(-)(1). Antimicrob Agents Chemother, 2000 Jun, 44(6), 1728 - 30 Efficacy of FK463, a (1,3)-beta-D-glucan synthase inhibitor, in disseminated azole-resistant candida albicans infection in mice; Maesaki S et al.; The efficacy of FK463, a new (1,3)-beta-D-glucan synthase inhibitor, against azole-resistant Candida albicans strains has been studied . The MIC of FK463 was lower than those of azoles and amphotericin B against CDR1-expressing C26 and CaMDR-expressing C40 strains . All mice treated with FK463 (1 mg/kg) survived disseminated murine candidiasis . The fungal burden in the kidney after 6 days was markedly reduced after therapy with FK463 and amphotericin B sodium deoxycholate, and plasma (1,3)-beta-D-glucan concentration was found to be lower in FK463-treated mice . In our study, FK463 was found to be a potent antifungal agent against disseminated infection with azole-resistant C . albicans. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1598 - 603 Mild heating of amphotericin B-desoxycholate: effects on ultrastructure, in vitro activity and toxicity, and therapeutic efficacy in severe candidiasis in leukopenic mice; van Etten EW et al.; Heated (20 min at 70 degrees C) amphotericin B-desoxycholate (hAMB-DOC) was further characterized, as was another formulation obtained after centrifugation (60 min, 3000 x g), hcAMB-DOC . Conventional AMB-DOC consisted of individual micelles (approximately 4 nm in diameter) and threadlike aggregated micelles, as revealed by cryo-transmission electron microscopy . For both hAMB-DOC and hcAMB-DOC, pleiomorphic cobweb structures were observed with a mean particle size of approximately 300 nm as determined by laser diffraction . The potent antifungal activity of AMB-DOC against Candida albicans is not reduced by heating . Effective killing of C . albicans (>99.9% within 6 h) was obtained at 0.1 mg/liter with each of the AMB formulations . For AMB-DOC, hAMB-DOC, and hcAMB-DOC, cation release ((86)Rb(+)) from C . albicans of > or =50% was observed at 0.8, 0.4, and 0.4 mg/liter, respectively . After heating of AMB-DOC, toxicity was reduced 16-fold as determined by red blood cell (RBC) lysis . For AMB-DOC, hAMB-DOC, and hcAMB-DOC, hemolysis of > or =50% was observed at 6.4, 102.4, and 102.4 mg/liter, respectively . In contrast, AMB-DOC and its derivates showed similar toxicities in terms of cation release from RBC . For AMB-DOC, hAMB-DOC, and hcAMB-DOC, cation release ((86)Rb(+)) of > or =50% was observed at 1.6, 0.8, and 0.8 mg/liter, respectively . In persistently leukopenic mice with severe invasive candidiasis, higher dosages of both hAMB-DOC and hcAMB-DOC were tolerated than those of conventional AMB-DOC (3 versus 0.8 mg/kg of body weight, respectively), resulting in significantly improved therapeutic efficacy . In conclusion, this new approach of heating AMB-DOC may be of great value for further optimizing the treatment of severe fungal infections. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1585 - 7 Does long-term itraconazole prophylaxis result in in vitro azole resistance in mucosal Candida albicans isolates from persons with advanced human immunodeficiency virus infection? The National Institute of Allergy and Infectious Diseases Mycoses study group; Goldman M et al.; The effects of prolonged itraconazole exposure on the susceptibility of Candida albicans isolates to itraconazole and fluconazole have not been well characterized . A recent placebo-controlled study of long-term itraconazole antifungal prophylaxis in persons with advanced human immunodeficiency virus infection afforded the opportunity to address this question . Mucosal Candida sp . isolates were obtained from subjects who developed oropharyngeal or esophageal candidiasis, and in vitro susceptibilities of the last isolate obtained at removal from the study as a prophylaxis failure were compared in itraconazole and placebo recipients . More subjects in the placebo group (74 of 146 {51%}) than in the itraconazole group (51 of 149 {34%}) developed mucosal candidiasis (P = 0.004) . A total of 112 isolates were recovered from 56 of the 74 (76%) subjects with mucosal candidiasis assigned to the placebo group, compared to 97 isolates from 45 of the 51 (88%) subjects in the itraconazole group . C . albicans accounted for 98% of isolates in the placebo group and 89% of isolates in the itraconazole group . The itraconazole MIC at which 50% of the isolates tested were inhibited (MIC(50)) for last-episode isolates from the itraconazole group was 0.125 microg/ml compared to 0.015 microg/ml for the placebo group subjects, P = 0.0001 . The MIC(50) of fluconazole for the last isolates from the itraconazole group was 1.5 microg/ml compared to 0.5 microg/ml for the placebo subjects (P = 0.005) . A lower proportion of isolates recovered from subjects on itraconazole therapy were classified as susceptible to itraconazole (63%) compared to isolates from the placebo group (96%) (P = 0.001) . Similarly, a lower proportion of C . albicans isolates from subjects on itraconazole therapy were susceptible to fluconazole (78%) compared to isolates from the placebo group (96%) (P = 0.01) . Also, the proportion of isolates that were not fully susceptible to itraconazole or fluconazole was greater in patients assigned to the itraconazole group than the placebo group (itraconazole susceptibility, 37 and 4%, respectively (P = 0.001); fluconazole susceptibility, 23 and 4%, respectively (P = 0.01) . In conclusion, long-term itraconazole prophylaxis in patients with AIDS is associated with reduction in susceptibility to itraconazole and cross-resistance to fluconazole. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1463 - 9 Antifungal activity of amphotericin B cochleates against Candida albicans infection in a mouse model; Zarif L et al.; Cochleates are lipid-based supramolecular assemblies composed of natural products, negatively charged phospholipid, and a divalent cation . Cochleates can encapsulate amphotericin B (AmB), an important antifungal drug . AmB cochleates (CAMB) have a unique shape and the ability to target AmB to fungi . The minimal inhibitory concentration and the minimum lethal concentration against Candida albicans are similar to that for desoxycholate AmB (DAMB; Fungizone) . In vitro, CAMB induced no hemolysis of human red blood cells at concentrations of as high as 500 microg of AmB/ml, and DAMB was highly hemolytic at 10 microg of AmB/ml . CAMB protect ICR mice infected with C . albicans when the agent is administered intraperitoneally at doses of as low as 0.1 mg/kg/day . In a tissue burden study, CAMB, DAMB, and AmBisome (liposomal AmB; LAMB) were effective in the kidneys, but in the spleen CAMB was more potent than DAMB at 1 mg/kg/day and was equivalent to LAMB at 10 mg/kg/day . In summary, CAMB are highly effective in treating murine candidiasis and compare well with AmBisome and AmB. Med Mycol, 2000 Apr, 38(2), 161 - 8 Antifungal activity of splenic, liver and pulmonary macrophages against Candida albicans and effects of macrophage colony-stimulating factor; Roilides E et al.; Disseminated infections due to Candida albicans are frequently encountered in immunocompromised patients . We compared the antifungal activities of macrophages residing in spleen, liver and lungs of rabbits against blastoconidia and pseudohyphae of C . albicans . Splenic adherent cells (SAC), Kupffer cells (KC) and pulmonary alveolar macrophages (PAM) all ingested blastoconidia efficiently . SAC caused significantly more damage to unopsonized pseudohyphae compared with KC (P < 0.01) or PAM (P < 0.001) . Incubation of SAC with 15 ng ml(-1) of recombinant human macrophage colony-stimulating factor (M-CSF) at 37 degrees C for 2 days significantly enhanced phagocytosis (P = 0.02) and killing (P = 0.05) of blastoconidia . In contrast, M-CSF had no effect on phagocytic activities of KC or PAM against blastoconidia or on damage caused by any of the macrophages to pseudohyphae of C . albicans . Thus, although all three resident macrophage types ingest blastoconidia efficiently, they differ in their capacity to cause damage to pseudohyphae and in their responsiveness to M-CSF for antifungal activation . M-CSF augments the capacity of SAC to ingest and kill blastoconidia and may therefore have a role in the treatment and prevention of hematogenously disseminated candidiasis. Med Mycol, 2000 Apr, 38(2), 155 - 9 An evaluation of the in vitro activity of terbinafine; Jessup CJ et al.; Terbinafine has previously been shown to be highly active against dermatophytes and many other filamentous fungi . However, its activity against yeasts is controversial, with earlier reports suggesting that it has low activity, while more recent studies demonstrated that terbinafine is effective against yeasts . In this study, the in vitro activity of terbinafine was evaluated against a broad range of fungal isolates . We examined the susceptibility of 100 yeast strains (10 species including Candida albicans, non-C . albicans, fluconazole-susceptible and -resistant candidal strains), and 184 strains of filamentous fungi and dermatophytes (29 species including Aspergillus, Fusarium, Sporothrix, Trichophyton rubrum, T . mentagrophytes, T . tonsurans, Microsporum canis and Epidermophyton floccosum), using the NCCLS M27-A microdilution methodology for yeasts and a modified M38-P methodology for moulds . The endpoint for terbinafine was defined as 80% inhibition compared with the growth control well . The mean yeast and filamentous fungi minimum inhibitory concentration values +/- SEM (in microg ml(-1)) for terbinafine were: 6.60 +/- 0.73 and 1.04 +/- 0.28, respectively . In conclusion, our data suggest that terbinafine, in addition to its potent activity against dermatophytes, is considerably effective against a broad range of yeasts and filamentous fungi in vitro . Therefore, investigations concerning its antifungal activity in vivo against such organisms should be pursued. Med Mycol, 2000 Apr, 38(2), 123 - 6 Identification of Candida albicans clinical isolates by PCR amplification of an EFB1 gene fragment containing an intron-interrupted open reading frame; Maneu V et al.; The use of a single pair of primers, deduced from the intron and exon nucleotide sequences of the Candida albicans EFB1 gene, in polymerase chain reaction (PCR) assays performed with whole cells of both laboratory strains and clinical isolates of Candida species, resulted in the species-specific amplification of a 785 bp DNA fragment in C . albicans strains . Clinical C . albicans isolates were tested, and 85 out of 86 generated the expected PCR-amplified product; other Candida species, both laboratory strains and clinical isolates, as well as laboratory strains belonging to other fungal genera, including medically relevant taxa, failed to amplify any DNA fragment . In addition, unusual C . albicans isolates (glucosamine- and N-acetylglucosamine-negative) from Africa also yielded the expected PCR-generated DNA fragment . These results indicate that genes containing intron sequences may be useful to design species-specific primers for the identification of fungal strains by PCR. Infect Immun, 2000 Jun, 68(6), 3485 - 90 Role of hyphal formation in interactions of Candida albicans with endothelial cells; Phan QT et al.; The ability to change from yeast to hyphal morphology is a major virulence determinant of Candida albicans . Mutants with defined defects in filamentation regulatory pathways have reduced virulence in mice . However, is it poorly understood why hyphal formation is critical for C . albicans to cause hematogenously disseminated infections . We used recently constructed mutants to examine the role of hyphal formation in the interactions of C . albicans with endothelial cells in vitro . These interactions included the ability of the mutants to invade and injure endothelial cells . Because the formation of hyphae may influence the host inflammatory response to C . albicans, we also investigated the capacity of these mutants to stimulate endothelial cells to express E-selectin and intercellular adhesion molecule 1 . We infected endothelial cells with C . albicans strains containing homozygous null mutations in the following filamentation regulatory genes: CLA4, CPH1, EFG1, and TUP1 . Whereas the wild-type strain formed true hyphae on endothelial cells, we found that neither the Deltaefg1 nor the Deltacph1 Deltaefg1 double mutant germinated . The Deltatup1 mutant formed only pseudohyphae . We also found that the Deltaefg1, Deltacph1 Deltaefg1, and Deltatup1 mutants had significantly reduced capacities to invade and injure endothelial cells . Therefore, Efg1p and Tup1p contribute to virulence by regulating hyphal formation and the factors that enable C . albicans to invade and injure endothelial cells . With the exception of the Deltacph1 Deltaefg1 mutant, all other mutants stimulated endothelial cells to express at least one of the leukocyte adhesion molecules . Therefore, the combined activities of Cph1p and Efg1p are required for C . albicans to stimulate a proinflammatory response in endothelial cells. Infect Immun, 2000 Jun, 68(6), 3297 - 304 Local anticandidal immune responses in a rat model of vaginal infection by and protection against Candida albicans; de Bernardis F et al.; Humoral (antibody {Ab}) and cellular Candida-specific immune responses in the vaginas of pseudoestrus rats were investigated during three successive infections by Candida albicans . After the first, protective infection, Abs against mannan and aspartyl proteinase antigens were present in the vaginal fluid, and their titers clearly increased during the two subsequent, rapidly healing infections . In all animals, about 65 and 10% of vaginal lymphocytes (VL) were CD3(+) (T cells) and CD3(-) CD5(+) (B cells), respectively . Two-thirds of the CD3(+) T cells expressed the alpha/beta and one-third expressed the gamma/delta T-cell receptor (TCR) . This proportion slightly fluctuated during the three rounds of C . albicans infection, but no significant differences between infected and noninfected rats were found . More relevant were the changes in the CD4(+)/CD8(+) T-cell ratio, particularly for cells bearing the CD25 (interleukin-2 receptor alpha) marker . In fact, a progressively increased number of both CD4(+) alpha/beta TCR and CD4(+) CD25(+) VL was observed after the second and third Candida challenges, reversing the high initial CD8(+) cell number of controls (estrogenized but uninfected rats) . The CD3(-) CD5(+) cells also almost doubled from the first to the third infection . Analysis of the cytokines secreted in the vaginal fluid of Candida-infected rats showed high levels of interleukin 12 (IL-12) during the first infection, followed by progressively increasing amounts of IL-2 and gamma interferon during the subsequent infections . No IL-4 or IL-5 was ever detected . During the third infection, VL with in vitro proliferative activity in response to an immunodominant mannoprotein antigen of C . albicans were present in the vaginal tissue . No response to this antigen by mitogen-responsive blood, lymph node, and spleen cells was found . In summary, the presence of protective Ab and T helper type 1 cytokines in the vaginal fluids, the in vitro proliferation of vaginal lymphocytes in response to Candida antigenic stimulation, and the increased number of activated CD4(+) cells and some special B lymphocytes after C . albicans challenge constitute good evidence for induction of locally expressed Candida-specific Ab and cellular responses which are potentially involved in anticandidal protection at the vaginal level. Infect Immun, 2000 Jun, 68(6), 3172 - 9 Characterization of binding of Candida albicans to small intestinal mucin and its role in adherence to mucosal epithelial cells; de Repentigny L et al.; In order to approximate and adhere to mucosal epithelial cells, Candida must traverse the overlying mucus layer . Interactions of Candida species with mucin and human buccal epithelial cells (BECs) were thus investigated in vitro . Binding of the Candida species to purified small intestinal mucin showed a close correlation with their hierarchy of virulence . Significant differences (P < 0.05) were found among three categories of Candida species adhering highly (C . dubliniensis, C . tropicalis, and C . albicans), moderately (C . parapsilosis and C . lusitaniae) or weakly (C . krusei and C . glabrata) to mucin . Adherence of C . albicans to BECs was quantitatively inhibited by graded concentrations of mucin . However, inhibition of adherence was reversed by pretreatment of mucin with pronase or C . albicans secretory aspartyl proteinase Sap2p but not with sodium periodate . Saturable concentration- and time-dependent binding of mucin to C . albicans was abrogated by pronase or Sap2p treatment of mucin but was unaffected by beta-mercaptoethanol, sodium periodate, neuraminidase, lectins, or potentially inhibitory sugars . Probing of membrane blots of the mucin with C . albicans revealed binding of the yeast to the 66-kDa cleavage product of the 118-kDa C-terminal glycopeptide of mucin . Although no evidence was found for the participation of C . albicans cell surface mannoproteins in specific receptor-ligand binding to mucin, inhibition of binding by p-nitrophenol (1 mM) and tetramethylurea (0.36 M) revealed that hydrophobic interactions are involved in adherence of C . albicans to mucin . These results suggest that C . albicans may both adhere to and enzymatically degrade mucins by the action of Saps, and that both properties may act to modulate Candida populations in the oral cavity and gastrointestinal tract. Phytother Res, 2000 May, 14(3), 207 - 9 Anticandidal activity of certain South Indian medicinal plants; Vaijayanthimala J et al.; The anticandidal activity of 20 household South Indian medicinal plants and/or plant products was studied using 30 Candida albicans isolates obtained from vaginal candidiasis patients of Rajah Muthiah Medical College and Hospital and compared with the anticandidal activity of garlic . Water and ethanol extracts were prepared and their minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC) were determined . Water extracts of three plants did not show any anticandidal activity, while Murrya koenigii did not exhibit any anticandidal activity in either extract . Other plants exhibited more activity in ethanol extracts showing that their active principle is more soluble in a non-polar solvent . Cell Biochem Funct, 2000 Jun, 18(2), 117 - 26 Antimicrobial and biological effects of ipemphos and amphos on bacterial and yeast strains; Ozturk AI et al.; In this study, the antimicrobial effects of monophosphazenes such as SM ipemphos and amphos were examined on bacterial and yeast strains . In addition, the biological effects of these compounds were tested on the lipid level of Saccharomyces cerevisiae and Candida albicans cells . The SM has an antimicrobial effect on the bacterial and yeast strains within the range of 100 and 1500 microg . When the concentration was increased, the inhibition zone expanded on the growth media ( p < 0.01; p < 0.001) . The ipemphos did not affect the bacterial and yeast cells in the 100 and 600 microg range . In addition, the amphos did not show an antimicrobial effect on the bacterial cells between 100 and 300 microg or on yeast cells at any of the administered concentrations . In vitro media, the biological effects of these molecules were compared with vitamin E, melatonin and fish oil on the yeast cells . We have found that monophosphazenes have growth effects on the cells in vitro media . The lipid level of S . cerevisiae cells was decreased by 300 microg doses of vitamin E, fish oil, and ipemphos (respectively; p < 0.05, p < 0.01, and p < 0 . 001) . In addition, the lipid levels of the same yeast cells were depressed by 1000-microg doses in all supplemented groups . However, it was observed that the highest decrease in lipid level of S . cerevisiae cells occurred in the amphos group ( p < 0.001) . The lipid levels of the C . albicans cells were significantly reduced ( p < 0.01) by 300 microg of amphos and melatonin . In contrast, the vitamin E and fish oil significantly raised ( p < 0.01; p < 0.001) the lipid level of the same yeast cell, as compared with the control . In addition, the lipid level of these cells was increased by administration of 1000 microg vitamin E, and melatonin ( p < 0.01) . In conclusion, while high concentrations of ipemphos and amphos have an antimicrobial effect on bacterial and yeast cells, amphos did not affect the yeast cells . While ipemphos and amphos increased cell growth in media, they reduced the lipid level of C . albicans and S . cerevisiae . In addition, the antioxidants such as vitamin E, melatonin, and fish oils affected the lipid level of yeast cells . Eur J Cardiothorac Surg, 2000 May, 17(5), 608 - 13 Pumpless extracorporeal lung assist - experience with the first 20 cases; Liebold A et al.; OBJECTIVE: Long-term extracorporeal lung assist is limited by a significant mechanical blood trauma resulting in bleeding and hemolysis . To reduce the drawbacks of extracorporeal lung assist a new technique has been developed, by which the driving force for the extracorporeal circuit derives from the patients arterio-venous pressure gradient (pumpless extracorporeal lung assist) . The aim of this clinical study was to test the feasibilty and effectiveness of pumpless extracorporeal lung assist in patients with acute respiratory distress syndrome . METHODS: Twenty patients (41+/-16 years) with acute respiratory distress syndrome of various causes and failing respirator therapy were enrolled . The minimum hemodynamic requirements included a cardiac output (CO) >6 l/min and mean arterial pressure (MAP) >70 mmHg . Pumpless extracorporeal lung assist was established using a short circuit arterio-venous shunt including a special designed low-resistance membrane oxygenator which was placed between the patients legs . RESULTS: At the time of inclusion FiO(2) in all patients was 1.0 (paO(2) 45.9+/-7 mmHg, paCO(2) 58.9+/-17 mmHg) . After 24 h of pumpless extracorporeal lung assist FiO(2) was reduced to 0.8+/-0.1 . A significant improvement in oxygenation (paO(2) 84.1+/-21 mmHg, P<0.05) and CO(2) removal (paCO(2) 32.7+/-5 mmHg, P<0.05) was notable . The mean extracorporeal flow was 2.6+/-0.6 l/min, which represented approximately 25% of the patients mean CO (10.8+/-2 l/min) . The median assist time was 12+/-8 (1-32) days . Fifteen out of twenty patients were weaned off pumpless extracorporeal lung assist . Five out of twenty patients died while on the system (four sepsis, one ventricular fibrillation) . Three out of twenty patients died after successful weaning on day 8, 30, and 50, respectively . Twelve out of twenty patients were discharged in a healthy state (overall survival 60%) . Technical problems included thrombosis of the venous cannula (n=5), thrombus formation within the membrane oxygenator (n=2), membrane oxygenator plasma leakage (n=2), and membrane oxygenator contamination with Candida albicans . No bleeding complication was observed . CONCLUSION: Pumpless extracorporeal lung assist is feasible and effective in a selected group of patients with acute respiratory distress syndrome but preserved hemodynamic function . By eliminating the pump and reducing the tubing length blood trauma can be minimized . Being very simple the system entails fewer risks of technical complications and also facilitates nursing care. Biotechnol Appl Biochem, 2000 Jun, 31 ( Pt 3), 213 - 8 Purification of native enolase from medically important Candida species; Ballantyne DS et al.; The 48 kDa glycolytic enzyme, enolase, has been identified as an immunodominant antigen in Candida albicans infections . It has also been identified as an important fungal allergen . Enolase from a number of medically important Candida species has been purified using a two-step anion- and cation-exchange chromatography method that was preceded by an organic extraction . The enolases purified by this method have a high specific activity and the procedure is 40% efficient, with an average of 5 mg of enolase/g of Candida cells . The purification of native enolase from medically important Candida species will enable the immunological significance and interspecies relationships of this major fungal antigen to be investigated. Proc Natl Acad Sci U S A, 2000 May 23, 97(11), 6102 - 7 Differential activation of a Candida albicans virulence gene family during infection; Staib P et al.; The yeast Candida albicans is a harmless commensal in most healthy people, but it causes superficial as well as life-threatening systemic infections in immunocompromised patients . C . albicans can colonize or infect virtually all body sites because of its high adaptability to different host niches, which involves the activation of appropriate sets of genes in response to complex environmental signals . We have used an in vivo expression technology that is based on genetic recombination as a reporter of gene expression to monitor the differential activation of individual members of a gene family encoding secreted aspartic proteinases (Saps), which have been implicated in C . albicans virulence, at various stages of the infection process . Our results demonstrate that SAP expression depends on the type of infection, with different SAP isogenes being activated during systemic disease as compared with mucosal infection . In addition, the activation of individual SAP genes depends on the progress of the infection, some members of the gene family being induced immediately after contact with the host, whereas others are expressed only after dissemination into deep organs . In the latter case, the number of invading organisms determines whether induction of a virulence gene is necessary for successful infection . The in vivo expression technology allows the elucidation of gene expression patterns at different stages of the fungus-host interaction, thereby revealing regulatory adaptation mechanisms that make C . albicans the most successful fungal pathogen of humans and, at the same time, identifying the stage of an infection at which certain virulence genes may play a role. J Exp Med, 2000 May 15, 191(10), 1661 - 74 Dendritic cells discriminate between yeasts and hyphae of the fungus Candida albicans . Implications for initiation of T helper cell immunity in vitro and in vivo; d'Ostiani CF et al.; The fungus Candida albicans behaves as a commensal as well as a true pathogen of areas highly enriched in dendritic cells, such as skin and mucosal surfaces . The ability of the fungus to reversibly switch between unicellular yeast to filamentous forms is thought to be important for virulence . However, whether it is the yeast or the hyphal form that is responsible for pathogenicity is still a matter of debate . Here we show the interaction, and consequences, of different forms of C . albicans with dendritic cells . Immature myeloid dendritic cells rapidly and efficiently phagocytosed both yeasts and hyphae of the fungus . Phagocytosis occurred through different phagocytic morphologies and receptors, resulting in phagosome formation . However, hyphae escaped the phagosome and were found lying free in the cytoplasm of the cells . In vitro, ingestion of yeasts activated dendritic cells for interleukin (IL)-12 production and priming of T helper type 1 (Th1) cells, whereas ingestion of hyphae inhibited IL-12 and Th1 priming, and induced IL-4 production . In vivo, generation of antifungal protective immunity was induced upon injection of dendritic cells ex vivo pulsed with Candida yeasts but not hyphae . The immunization capacity of yeast-pulsed dendritic cells was lost in the absence of IL-12, whereas that of hypha-pulsed dendritic cells was gained in the absence of IL-4 . These results indicate that dendritic cells fulfill the requirement of a cell uniquely capable of sensing the two forms of C . albicans in terms of type of immune responses elicited . By the discriminative production of IL-12 and IL-4 in response to the nonvirulent and virulent forms of the fungus, dendritic cells appear to meet the challenge of Th priming and education in C . albicans saprophytism and infections. J Biol Chem, 2000 May 19, 275(20), 14882 - 9 Glycosylphosphatidylinositol-anchored glucanosyltransferases play an active role in the biosynthesis of the fungal cell wall; Mouyna I et al.; A novel 1,3-beta-glucanosyltransferase isolated from the cell wall of Aspergillus fumigatus was recently characterized . This enzyme splits internally a 1,3-beta-glucan molecule and transfers the newly generated reducing end to the non-reducing end of another 1, 3-beta-glucan molecule forming a 1,3-beta linkage, resulting in the elongation of 1,3-beta-glucan chains . The GEL1 gene encoding this enzyme was cloned and sequenced . The predicted amino acid sequence of Gel1p was homologous to several yeast protein families encoded by GAS of Saccharomyces cerevisiae, PHR of Candida albicans, and EPD of Candida maltosa . Although the expression of these genes is required for correct morphogenesis in yeast, the biochemical function of the encoded proteins was unknown . The biochemical assays performed on purified recombinant Gas1p, Phr1p, and Phr2p showed that these proteins have a 1,3-beta-glucanosyltransferase activity similar to that of Gel1p . Biochemical data and sequence analysis have shown that Gel1p is attached to the membrane through a glycosylphosphatidylinositol in a similar manner as the yeast homologous proteins . The activity has been also detected in membrane preparations, showing that this 1,3-beta-glucanosyltransferase is indeed active in vivo . Our results show that transglycosidases anchored to the plasma membrane via glycosylphosphatidylinositols can play an active role in fungal cell wall synthesis. J Bacteriol, 2000 Jun, 182(11), 3063 - 71 Morphogenesis, adhesive properties, and antifungal resistance depend on the Pmt6 protein mannosyltransferase in the fungal pathogen candida albicans; Timpel C et al.; Protein mannosyltransferases (Pmt proteins) initiate O glycosylation of secreted proteins in fungi . We have characterized PMT6, which encodes the second Pmt protein of the fungal pathogen Candida albicans . The residues of Pmt6p are 21 and 42% identical to those of C . albicans Pmt1p and S . cerevisiae Pmt6p, respectively . Mutants lacking one or two PMT6 alleles grow normally and contain normal Pmt enzymatic activities in cell extracts but show phenotypes including a partial block of hyphal formation (dimorphism) and a supersensitivity to hygromycin B . The morphogenetic defect can be suppressed by overproduction of known components of signaling pathways, including Cek1p, Cph1p, Tpk2p, and Efg1p, suggesting a specific Pmt6p target protein upstream of these components . Mutants lacking both PMT1 and PMT6 are viable and show pmt1 mutant phenotypes and an additional sensitivity to the iron chelator ethylenediamine-di(o-hydroxyphenylacetic acid) . The lack of Pmt6p significantly reduces adherence to endothelial cells and overall virulence in a mouse model of systemic infection . The results suggest that Pmt6p regulates a more narrow subclass of proteins in C . albicans than Pmt1p, including secreted proteins responsible for morphogenesis and antifungal sensitivities. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2000 May, 89(5), 570 - 6 Insulin-dependent diabetes mellitus and oral soft tissue pathologies: II . Prevalence and characteristics of Candida and Candidal lesions; Guggenheimer J et al.; OBJECTIVE: To assess the prevalence of Candida albicans and oral infection with Candida in patients with insulin-dependent diabetes mellitus (IDDM).Study Design: This cross-sectional study compared the prevalence of candidiasis in 405 subjects with IDDM and 268 nondiabetic control subjects . Assessments included evidence of clinical manifestations of candidiasis and a quantitative measure of Candida pseudohyphae in a cytologic smear from the midline posterior dorsal tongue . RESULTS: More subjects with IDDM than control subjects without IDDM (15.1% vs 3.0%) were found to have clinical manifestations of candidiasis, including median rhomboid glossitis, denture stomatitis, and angular cheilitis . IDDM subjects were also more likely to have any Candida pseudohyphae in their cytologic smears (23.0% vs 5.7%; P <.0001), as well as pseudohyphae counts of >10/cm(2) (7.1% vs 0.8%; P <.0001) . Diabetic subjects with median rhomboid glossitis were more likely to have a longer duration of IDDM and complications of nephropathy and retinopathy . Denture stomatitis was associated with smoking, retinopathy, higher Candida counts, poor glycemic control, and longer duration of IDDM . A multivariate regression analysis found 3 factors to be significantly associated with the presence of Candida pseudohyphae in the subjects with IDDM: current use of cigarettes (odds ratio, 2:4), use of dentures (odds ratio, 2:3), and elevated levels of glycosylated hemoglobin (odds ratio, 1:9) . The use of antimicrobials, immunosuppressants, or drugs with xerostomic side effects was not related to the presence of Candida . CONCLUSIONS: Candida pseudohyphae and oral soft tissue manifestations of candidiasis were more prevalent in subjects with IDDM than in control subjects without diabetes . The presence of Candida pseudohyphae was significantly associated with cigarette smoking, use of dentures, and poor glycemic control. Science, 2000 May 12, 288(5468), 1062 - 4 A high-affinity iron permease essential for Candida albicans virulence; Ramanan N et al.; Microbial pathogens must compete with the iron-withholding defense systems of their host to acquire this essential nutrient . Here, two high-affinity iron permease genes, CaFTR1 and CaFTR2, were isolated . CaFTR1 expression was induced under iron-limited conditions and repressed when iron supply was sufficient, whereas the expression of CaFTR2 was regulated in a reversed manner . Mutants lacking CaFTR1 but not CaFTR2 exhibited a severe growth defect in iron-deficient medium and were unable to establish systemic infection in mice . Thus, CaFTR1-mediated iron-uptake mechanism constitutes a virulence factor of Candida albicans and may be a target for the development of anti-Candida therapies. Arch Otolaryngol Head Neck Surg, 2000 May, 126(5), 672 - 4 Adult herpetic laryngitis with concurrent candidal infection: a case report and literature review; Zhang S et al.; Rarely, adult herpetic laryngitis without involvement of the oropharynx has been reported . However, to our knowledge, laryngitis caused by herpes simplex virus with coexisting Candida albicans has not been reported . We report what we believe to be the first case of localized herpetic laryngitis superimposed by laryngeal Candida species infection in an immunosuppressed patient . This diagnosis was made on the basis of the findings of a laryngeal mucosal biopsy and ancillary testing using fungal stains and immunohistochemical stains for herpetic antigens . We also review the literature and discuss the clinical and diagnostic presentations, including potential pitfalls in the diagnosis. Arch Immunol Ther Exp (Warsz), 2000, 48(2), 101 - 5 Interleukin 6, tumor necrosis factor alpha and their soluble receptors in the blood serum of patients with denture stomatitis and fungal infection; Pietruski JK et al.; Determinations of the blood serum levels of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-alpha) and their soluble receptors (sIL-6R, sTNFR) in denture stomatitis patients (DS) were performed . Serum levels of interleukins and their soluble receptors were measured using the ELISA method . In all examined patients mycological diagnostics were conducted using API 20C AUX stripe tests and an automatic ATB machine . Results were compared with those of healthy denture wearers (D), and controls (C) . In DS patients, yeasts were isolated in 90.9%, in D in 66.7% of cases . The most often isolated species in both groups was Candida albicans . Mean concentrations of IL-6 and TNF-alpha were statistically significantly higher in DS and D groups compared to controls . Mean concentrations of sIL-6R were similar in all groups; however, concentrations of sTNFR in both DS and D groups were significantly lower compared to controls . There were no correlations found between values of IL-6 and TNF-alpha nor between examined interleukins and their soluble receptors. Eur J Med Res, 2000 May 23, 5(5), 203 - 8 Treatment of candidal infections with fluconazole in neonates and infants; Schwarze R et al.; To study the therapy, efficacy and safety of fluconazole in candidal mycoses during neonatal phase and infancy a case review in 53 newborns and infants was performed . The majority of these patients were premature with a median birth weight of 1120 g and born within gestational week 23-38 . The median age at the onset of fluconazole treatment was 5 weeks . All patients had underlying diseases and several risk factors, which favored the occurence of a systemic candidal mycosis . Systemic candidiasis was the most frequent diagnosis (75.5%) . Fluconazole was administered at a daily dosage of 5-6 mg/kg for a median duration of 21 days . The hepatic, renal and hematologic functions were assessed before, one, two, and three weeks after start of treatment . Yeasts were identified in 37 patients . The most common fungus isolated at baseline was Candida albicans (68%) . Clinical cure or improvement was reported in 31 out of 38 patients (81.6%) . Mycological cure was achieved in 25 out of 32 newborns and infants . Despite the limited number of patients with outcome data, these preliminary results of a small cohort clearly indicate the effective antifungal therapy with fluconazole in neonates and infants . No serious side effects were observed in fluconazole-treated patients . Two patients with megaureter-megacystis-hydronephrosis syndrome and severe meningoencephalitis showed a mild increase in liver enzymes . - Conclusion: Fluconazole seems to be an effective therapy for systemic and other forms of candidiasis in infants including very low birth weight infants (VLBWI; <1500 g) . These favorable safety and efficacy data are similar to results obtained with fluconazole in older children and adults . These findings, however, must be supported by larger trials . The recommended daily dose is 5 mg/kg body weight . Only in VLBWI the dosage interval within the first two weeks of life should be prolonged up to 3 days and fluconazole serum levels should be monitored. J Antibiot (Tokyo), 2000 Feb, 53(2), 158 - 62 Enhancement of the antifungal activity of rapamycin by the coproduced elaiophylin and nigericin; Fang A et al.; Streptomyces hygroscopicus ATCC 29253 produces rapamycin, elaiophylin and nigericin . Although elaiophylin has no activity against Candida albicans ATCC 11651, it markedly enhances rapamycin's antifungal activity . Nigericin has only weak activity on its own but it also enhances rapamycin action . Surprisingly, elaiophylin does not enhance nigericin activity on C . albicans. Cytokine, 2000 Apr, 12(4), 379 - 87 Interleukin 10 suppresses phagocytic and antihyphal activities of human neutrophils; Roilides E et al.; We investigated the effects of human interleukin 10 (IL-10) on the antibacterial and antifungal activities of human neutrophils (PMNs) against Staphylococcus aureus and Candida albicans . Incubation of PMNs from healthy volunteers with 20-100 ng/ml of IL-10 at 37 degrees C for 1 h suppressed phagocytosis of serum-opsonized S . aureus (P=0.02) and blastoconidia of C . albicans (P<0.01) . In contrast, 2-100 ng/ml of IL-10 had no effect on superoxide anion production upon stimulation with phorbol myristate acetate, N-formylmethionyl leucyl phenylalanine, C . albicans blastoconidia or pseudohyphae; neither did it significantly affect conidiocidal or bactericidal activities of PMNs . However, 20-100 ng/ml of IL-10 significantly decreased PMN-induced damage of C . albicans pseudohyphae (P=0.008) . The suppression of phagocytic activity of PMNs against S . aureus and blastoconidia of C . albicans as well as the impairment of PMN-induced hyphal damage may have important implications for understanding the immunosuppressive profile of IL-10 in clinical usage. J Reprod Med, 2000 Apr, 45(4), 354 - 6 Candida albicans chorioamnionitis associated with preterm labor and sudden intrauterine demise of one twin . A case report; Friebe-Hoffmann U et al.; BACKGROUND: Although cervicovaginal Candida infections occur in 20-25% of pregnancies, the incidence of ascending infection in these cases is only 0.8%, and such infection rarely causes chorioamnionitis . CASE: Sudden intrauterine fetal demise (IUFD) of twin A occurred in a diabetic primigravida presenting with a twin pregnancy and preterm labor at 33 weeks of gestation . Placental pathology and autopsy of the stillborn twin revealed extensive chorioamnionitis and fetal sepsis in the presence of Candida albicans . Twin B was unaffected . CONCLUSION: In this case, C albicans chorioamnionitis seemed to be associated with sudden IUFD. FEMS Microbiol Lett, 2000 May 15, 186(2), 239 - 43 Evidence for the presence of pir-like proteins in Candida albicans; Kandasamy R et al.; Pir proteins are unique proteins with internal repeat sequences that are reported to be present in the cell wall of Saccharomyces cerevisiae . They are covalently attached to the cell wall and can be released by mild alkali treatment . In this study the biotinylated cell wall preparations from Candida albicans and S . cerevisiae were extracted by alkali and beta-1,3 glucanase and analyzed in parallel . Among the four bands detected by streptavidin, two proteins were recognized by the antibody to the S . cerevisiae Pir protein Hsp150 . The antibody also detected a high molecular mass protein secreted in the growth medium of C . albicans . Using S . cerevisiae HSP150/PIR2 gene as a probe, Southern and Northern hybridizations were performed with DNA and RNA of C . albicans . Hybridization with DNA digested with different restriction enzymes showed more than one hybridized fragment . An increased level of mRNA was found in heat shocked cells (37 degrees C for 45 min compared to 25 degrees C) . Hybridization of ScHSP150 gene to mRNAs from cells grown in different media was also determined . Two transcripts of size approximately 3.5 kb and 2.0 kb were detected in mRNAs from cells grown in defined medium with glucose as carbon source or in the same medium supplemented with hemoglobin . The lower transcript of size 2.0 kb was absent in cells grown in medium with galactose as carbon source . A single band was also observed when cells were grown in rich medium . Together these results demonstrated the existence of beta1,3 glucan linked proteins in C . albicans, which are related to Pir family proteins of S . cerevisiae. J Oral Pathol Med, 2000 May, 29(5), 206 - 13 Sub-therapeutic exposure to polyene antimycotics elicits a post-antifungal effect (PAFE) and depresses the cell surface hydrophobicity of oral Candida albicans isolates; Egusa H et al.; Post-antifungal effect (PAFE) is defined as the suppression of growth that persists following limited exposure of fungi to antimycotics and subsequent removal of the drug . The fungal pathogen Candida albicans is the major aetiologic agent of oral candidosis, and the cell surface hydrophobicity (CSH) of this yeast is considered a critical factor contributing to its colonisation potential . As the concentration of topically prescribed antifungals reach sub-therapeutic levels at dosage intervals, the study of the polyene-induced PAFE and its impact on the CSH of oral C . albicans should be of clinical relevance . Hence the aims of this investigation were to measure the PAFE and CSH of 12 isolates of C . albicans following limited exposure (1 h) to nystatin and amphotericin B and also to investigate the ultrastructural features of yeast cells following such antifungal exposure . The yeasts were exposed to sub-lethal concentrations of nystatin (x2 MIC) and amphotericin B (x2 MIC) for a period of 1 h . Following subsequent removal of the drug, the PAFE and the CSH of the isolates were assessed by a turbidometric measurement of growth and a biphasic aqueous-hydrocarbon assay, respectively . The mean duration of PAFE of nystatin and amphotericin B were 5.99 (+/-0.49) h and 8.73 (+/-0.93) h, respectively, while the reduction in CSH following exposure to these drugs were 17.32% (P<0.05 for 83% of the isolates) and 14.26% (P<0.05 for 66% of the isolates), respectively . On scanning electron microscopy the exposed cells were seen to undergo collapse of the internal cell membrane, leaving an intact cell wall, while a proportion of cells were deflated . Some cells showed intense puckering of the cell wall, resulting in a mulberry appearance . Taken together, these data elucidate additional mechanisms by which polyene antimycotics may operate in vivo to suppress candidal pathogenicity. J Oral Pathol Med, 2000 May, 29(5), 193 - 9 Biotypes of oral Candida albicans isolated from AIDS patients and HIV-free subjects in Thailand; Teanpaisan R et al.; This study was conducted to examine biotypes and antifungal susceptibility patterns of oral Candida albicans isolated from HIV-infected patients, HIV-free patients with candidiasis and healthy subjects . All isolates were biotyped using a typing system based on enzyme profiles, carbohydrate assimilation patterns and boric acid resistance . Thirty-eight biotypes were found amongst 218 oral C . albicans isolates . The major biotype found was A1S, which accounted for 32.6% of all isolates, and this biotype was the most common in all groups . There was a greater variety of biotypes of C . albicans in the HIV-infected group than in the other groups; however, there was no statistically significant difference between the groups . The minimum inhibitory concentrations (MICs) of a total of 118 isolates were determined for amphotericin B and for ketoconazole using the National Committee for Clinical Laboratory Standards (NCCLS) broth macrodilution method and the E-test . When the antifungal susceptibility patterns among the groups were compared, a statistically significant difference was found only with amphotericin B . The median MIC of amphotericin B in the HIV-infected group was higher than in the healthy group (P=0.013, NCCLS method; P=0.002, E-test) . However, this difference in sensitivity was not restricted to any sub-type investigated . Our results showed that the biotype patterns of C . albicans isolates that colonize HIV-infected patients are similar to those of HIV-free subjects, and there is no relationship between antifungal susceptibility patterns and the biotypes. Indian J Pediatr, 1999 Jan-Feb, 66(1), 85 - 91 Prevalence of intestinal pathogens in HIV patients with diarrhea: implications for treatment; Ramakrishna BS; Patients infected with the human immunodeficiency virus (HIV) commonly experience diarrhea at some time during their illness . A variety of enteric pathogens are identified in 50-80% of these patients, depending on the intensity of the diagnostic work-up that is done . In addition to the common enteric pathogens, several unusual enteric pathogens are recognized to cause diarrhea especially in HIV patients . These include protozoan parasites such as Cryptosporidia, Isospora belli, Cyclospora cayatenensis and Microsporidium species bacteria such as enteropathogenic Escherichia coli and Mycobacterium avium-intracellulare, fungi including Candida albicans and Histoplasma capsulatum, and viruses such as astroviruses and caliciviruses . Diagnosis of these infections sometimes involves special procedures not readily available every where, and empiric therapy based on knowledge of the likely pathogens has been advocated for developing countries . This article reviews the currently available data on geographic variation of enteric pathogens in HIV patients with diarrhea and outlines a rational strategy for empiric therapy of these patients. Clin Chem, 2000 May, 46(5), 631 - 5 Rapid detection of point mutations by fluorescence resonance energy transfer and probe melting curves in Candida species; Loeffler J et al.; BACKGROUND: The LightCycler(TM) combines rapid amplification of nucleic acids in glass capillaries with melting curve analysis based on fluorescence resonance energy transfer for the sensitive detection of point mutations in various settings, such as drug resistance and hereditary diseases . Point mutations leading to an altered structure of lanosteroldemethylase, the target enzyme of the fungistatic azoles, are an important mechanism of acquired resistance in Candida albicans . METHODS: We screened 13 fluconazole-resistant C . albicans and 21 fluconazole-resistant C . tropicalis strains (minimum inhibitory concentration >128 mg/L), isolated from patients with AIDS, for the presence of defined point mutations by comparing conventional cycle sequencing with a newly designed LightCycler-based assay . RESULTS: In C . tropicalis, 5 of 21 isolates showed the wild-type sequence, and 8 of 21 showed the homozygous nucleotide exchange thymine to cytosine at position 1554 (T1554C) . A heterozygous genotype was detected in 8 of 21 isolates by the LightCycler, but in only 3 of 21 isolates by conventional cycle sequencing . In 2 of 13 C . albicans isolates, a homozygous point mutation leading to an amino acid exchange at position 464 (glycine to serine) was detected in both assays . CONCLUSION: The LightCycler technique offers standardized, fast, sensitive, and reproducible detection of point mutations in different Candida spp. IUBMB Life, 1999 Aug, 48(2), 163 - 8 DNA-dependent RNA polymerase II from Candida species is a multiple zinc-containing metalloenzyme; Patturajan M et al.; We have purified DNA-dependent RNA polymerase II from Candida albicans, a human pathogenic yeast . The enzyme consists of 9 polypeptides that are unique to C . albicans, their mobility on SDS-PAGE being different from the mobility of the corresponding subunits of RNA polymerase II from Saccharomyces cerevisiae or C . utilis . In the present study we also demonstrate that RNA pol II from C . albican and C . utilis are metalloproteins containing approximately 5 mol of zinc per mole of enzyme . Although prolonged dialysis in 10 or 20 mM EDTA failed to remove Zn(II) from the C . albicans enzyme, in the C . utilis enzyme 3 Zn(II) ions could be removed and then reconstituted in the presence of excess Zn(II) . o-Phenanthroline (5 mM) removed Zn(II) from C . albicans enzyme irreversibly in a time-dependent fashion with concomitant loss of enzyme activity . Circular dichroism studies revealed structural changes on removal of zinc, thus suggesting a role for Zn in maintenance of structural stability . Further, we demonstrate that the largest subunit of the C . utilis enzyme and the 3 large subunits of the C . albicans enzyme can bind radioactive zinc. Mol Microbiol, 2000 Apr, 36(2), 290 - 301 Aspergillus SteA (sterile12-like) is a homeodomain-C2/H2-Zn+2 finger transcription factor required for sexual reproduction; Vallim MA et al.; Saccharomyces cerevisiae Ste12p plays a key role in coupling signal transduction through MAP kinase modules to cell-specific or morphogenesis-specific gene expression required for mating and pseudohyphal (PH)/filamentous growth (FG) . Ste12p homologues in the pathogenic yeasts Candida albicans and Filobasidiela neoformans apparently play similar roles during dimorphic transitions . Here we report the isolation and characterization of the first Ste12 protein from a true filamentous fungus . Aspergillus nidulans steA encodes a protein with a homeodomain 63-75% identical to those of other Ste12 proteins, with greatest similarity to FnSte12alphap . SteAp and Ste12alphap lack the pheromone induction domain found in budding yeast Ste12p, but have C-terminal C2/H2-Zn+2 finger domains not present in the other Ste12 proteins . A DeltasteA strain is sterile and differentiates neither ascogenous tissue nor fruiting bodies (cleistothecia) . However, the development of sexual cycle-specific Hulle cells is unaffected . Filamentous growth, conidiation and the differentiation of PH-like asexual reproductive cells (metulae and phialides) are normal in the deletion strain . Northern analysis of key regulators of the asexual and sexual reproductive cycles support the observation that although SteAp function is restricted to the sexual cycle, cross regulation between the two developmental pathways exists . Our results further suggest that while several classes of related proteins control similar morphogenetic events in A . nidulans and the dimorphic yeasts, significant differences must exist in the regulatory circuitry. Am J Vet Res, 1999 Oct, 60(10), 1255 - 61 Analysis of surface antigen expression and host defense function in leukocytes from calves heterozygous or homozygous for bovine leukocyte adhesion deficiency; Sipes KM et al.; OBJECTIVE: To analyze surface antigen expression and functional responses of leukocytes from calves heterozygous and homozygous for bovine leukocyte adhesion deficiency (BLAD) . ANIMALS: 8 clinically normal calves, 4 calves heterozygous for BLAD, and 4 calves homozygous for BLAD . PROCEDURE: Surface antigen expression was examined by flow cytometric analysis of leukocytes stained with monoclonal antibodies . Neutrophil function analyses included phagocytosis and killing of Candida albicans and measurement of respiratory burst activity using cytochrome c and dihydrorhodamine 123 assays . Differential leukocyte counts also were performed . RESULTS: Leukocytes from heterozygous calves were similar to those of clinically normal calves with respect to surface antigen expression, C albicans phagocytosis and killing, and respiratory burst activity . In contrast, neutrophils from calves homozygous for BLAD had significantly reduced phagocytic and yeast-killing capacity but had higher respiratory burst activity than cells from clinically normal or heterozygous calves . Homozygous calves also had extreme neutrophilia and significantly more immature neutrophils . CONCLUSIONS: The heterozygous BLAD genotype does not cause detectable functional differences in leukocytes, compared with those of clinically normal calves . In contrast, leukocytes from homozygous calves seem to upregulate alternative host defense capabilities (eg, respiratory burst activity) to partially compensate for the lack of typical adherence-dependent host defense functions. Genetics, 2000 May, 155(1), 57 - 67 TUP1, CPH1 and EFG1 make independent contributions to filamentation in candida albicans; Braun BR et al.; The common fungal pathogen, Candida albicans, can grow either as single cells or as filaments (hyphae), depending on environmental conditions . Several transcriptional regulators have been identified as having key roles in controlling filamentous growth, including the products of the TUP1, CPH1, and EFG1 genes . We show, through a set of single, double, and triple mutants, that these genes act in an additive fashion to control filamentous growth, suggesting that each gene represents a separate pathway of control . We also show that environmentally induced filamentous growth can occur even in the absence of all three of these genes, providing evidence for a fourth regulatory pathway . Expression of a collection of structural genes associated with filamentous growth, including HYR1, ECE1, HWP1, ALS1, and CHS2, was monitored in strains lacking each combination of TUP1, EFG1, and CPH1 . Different patterns of expression were observed among these target genes, supporting the hypothesis that these three regulatory proteins engage in a network of individual connections to downstream genes and arguing against a model whereby the target genes are regulated through a central filamentous growth pathway . The results suggest the existence of several distinct types of filamentous forms of C . albicans, each dependent on a particular set of environmental conditions and each expressing a unique set of surface proteins. Ear Nose Throat J, 2000 Apr, 79(4), 278 - 80, 282, 284-5 Aggressive combination treatment for invasive fungal sinusitis in immunocompromised patients; Rizk SS et al.; Invasive sinonasal fungal disease is a potentially fatal complication of chemotherapy-induced immunosuppression and neutropenia . We reviewed the outcomes of seven cancer patients who had been diagnosed with invasive fungal sinusitis; six patients had hematologic malignancies and one had breast cancer . At the time of their sinus diagnosis, all patients had been hospitalized and were receiving combination chemotherapy for their underlying malignancy . Impairment of their immune function was characterized by an absolute neutrophil count of less than 1,000/mm3 . Aggressive management of their sinonasal fungal disease consisted of surgical debridement and systemic amphotericin B for all patients, and treatment with granulocyte colony-stimulating factor for two patients . Invasive Aspergillus infection was identified in six patients and invasive Candida albicans infection in one . Although the prognosis for these patients was poor and two patients died of the fungal infection, the aggressive treatment strategy resulted in long-term survival for the remaining five patients. Folia Med (Plovdiv), 1999, 41(4), 68 - 72 Antifungal prophylaxis with low doses fluconazole in patients with hematological malignancies; Goranov S et al.; INTRODUCTION: The incidence of mycotic infections in immunocompromised patients has reached a 20-fold rise for the last two decades . AIM: The goal of the study was to evaluate fluconazole efficacy as antifungal prophylaxis in patients with hematological malignancies . MATERIAL AND METHODS: Sixty nine patients with hematological malignancies and neutrophil count less than 1.0 x 10(9)/L received fluconazole orally at a dose of 150 mg every other day . Fluconazole was discontinued when a neutrophil count above 1.5 x 10(9)/L was maintained . Duration of neutropenic periods, afebrile cycles, and incidence of mycotic infections were assessed . The same variables were observed in a control group of 41 patients who did not receive antifungal prophylactic therapy . RESULTS: Both groups were similar in the mean duration of neutropenic period but the afebrile cycles in the patients receiving antifungal prophylaxis were longer by 5 days compared to these in the patients without prophylaxis . The mycotic infections had lower incidence in the patients on antifungal prophylaxis and the difference was statistically significant (p < 0.01) . The superficial mycotic infections composed the majority of the clinically and/or microbiologically verified infections . These were presented by oropharyngeal infection (61%), esophageal (22%), and single cases of skin, genital or rectal infections . Candida albicans was isolated in 85% of the cases . The adverse reactions of fluconazole therapy were mild, transient, and easily manageable . No signs of liver and renal dysfunction were observed . CONCLUSION: Lower dose of fluconazole, 150 mg every other day p.o., has the same antifungal effect as a dose of 200 mg/day p.o . in neutropenic patients with hematological malignancies which lowers the cost of treatment. Microbiology, 2000 Apr, 146 ( Pt 4), 869 - 76 Candida albicans CFL1 encodes a functional ferric reductase activity that can rescue a Saccharomyces cerevisiae fre1 mutant; Hammacott JE et al.; Candida albicans, like other pathogens, has to compete with the host for a limited supply of available iron . Consequently, iron acquisition is likely to be an important factor for the growth, survival and virulence of this organism . It was previously demonstrated that C . albicans has a surface-associated ferric reductase similar to that of Saccharomyces cerevisiae . Therefore, functional rescue of a S . cerevisiae fre1 mutant was used to isolate a C . albicans ferric reductase gene (CFL1) . This gene has been previously identified . However, the workers had not observed any functional reductase activity associated with the gene . The discrepancy with the findings in this report appears to be due to the clone previously reported carrying a non-contiguous piece of C . albicans DNA . Results shown in this paper demonstrate that CFL1 transcription is regulated in response to levels of iron and copper . This is the first demonstration of a functional ferric reductase gene from C . albicans. Lipids, 2000 Mar, 35(3), 257 - 62 Cloning and sequencing of the Candida albicans C-4 sterol methyl oxidase gene (ERG25) and expression of an ERG25 conditional lethal mutation in Saccharomyces cerevisiae; Kennedy MA et al.; The ERG25 gene encoding the Candida albicans C-4 sterol methyl oxidase was cloned and sequenced by complementing a Saccharomyces cerevisiae erg25 mutant with a C . albicans genomic library . The Erg25p is comprised of 308 amino acids and shows 65 and 38% homology to the enzymes from S . cerevisiae and Homo sapiens, respectively . The protein contains three histidine clusters common to nonheme iron-binding enzymes and an endoplasmic reticulum retrieval signal as do the proteins from S . cerevisiae and humans . A temperature-sensitive (ts) conditional lethal mutation of the C . albicans ERG25 was isolated and expressed in S . cerevisiae . Sequence analysis of the ts mutant indicated an amino acid substitution within the region of the protein encompassed by the histidine clusters involved in iron binding . Results indicate that plasmid-borne conditional lethal mutants of target genes have potential use in the rescue of Candida mutations in genes that are essential for viability. Crit Rev Microbiol, 2000, 26(1), 59 - 68 The role of Candida dubliniensis in oral candidiasis in human immunodeficiency virus-infected individuals; Schorling SR et al.; There is an increasing interest in non-albicans Candida species because of the increasing number of fungal infections they cause . Most of these infections can be found in immunocompromised individuals, especially in those infected with human immunodeficiency virus (HIV) . Candida dubliniensis is a recently identified yeast, mostly isolated in HIV-positive individuals with oral candidiasis . Candida dubliniensis is a germ tube- and chlamydospore-form yeast . Thus, it shares diagnostic characteristics with Candida albicans . Probably, Candida dubliniensis has been present in the community for a long time and has been misidentified as Candida albicans . Significant phenotypic characteristics of Candida dubliniensis (difference in the carbohydrate assimilation profile, difference in colony color on CHROMagar Candida, and positive tetrazolium test, etc.) have been found, but none of them seem to be sufficient alone for the definitive identification of the species . Recently, PCR tests were developed to discriminate Candida albicans from Candida dubliniensis . However, these prove difficult in the context of routine mycological diagnostics . Moreover, an increased resistance to antifungal drugs has been described . This shows the importance of identification of Candida dubliniensis . To elucidate the current insight into Candida dubliniensis, the phenotypic and genotypic characteristics as well as the prevalence and the antifungal drug susceptibilities of this species are discussed from a clinical standpoint. Clin Immunol, 2000 May, 95(2), 145 - 55 Glycyrrhizin improves the resistance of MAIDS mice to opportunistic infection of Candida albicans through the modulation of MAIDS-associated type 2 T cell responses; Utsunomiya T et al.; Compared with normal mice, MAIDS mice (mice infected with LP-BM5 murine leukemia virus) exhibited an increase up to 100 times greater in susceptibility to infection with Candida albicans . The impaired resistance of MAIDS mice to the infection was recovered to levels observed in normal mice by the administration of glycyrrhizin (GR), an active component of licorice roots . MAIDS mice inoculated with CD4(+) T cells from GR-treated mice were also resistant to C . albicans infection . Normal mice inoculated with CD4(+) T helper type 2 cells (Th2 cells) from MAIDS mice were susceptible to C . albicans infection at the same levels shown in MAIDS mice . The susceptibility of normal mice inoculated with type 2 T cells was reversible by (i) administration of GR and (ii) inoculation of CD4(+) T cells from GR-treated mice and injection of a mixture of mAbs targeted against type 2 cytokines (IL-4 and IL-10) . Type 2 cytokines were not detected in sera of MAIDS mice inoculated with CD4(+) T cells from GR-treated mice, while they were present in sera of MAIDS mice treated with saline . These results suggest that, by inducing CD4(+) T cells which suppress type 2 cytokine production by MAIDS-associated Th2 cells, GR improves the resistance of MAIDS mice to C . albicans infection . Nippon Ishinkin Gakkai Zasshi, 2000, 41(2), 115 - 9 {Protective effect of oral administration of several traditional Kampo-medicines on lethal Candida infection in immunosuppressed mice}; Abe S et al.; The protective effects of a "hozai" type of Kampo medicine, Juzen-taiho-to (Shi-quan-da-bu-tang, TJ-48), Hochu-ekki-to (Bu-zhong-yi-qi-tang, TJ-41) or Ninjin-yoei-to (Ren-shen-yang-rong-tang, TJ-108) on experimental candidiasis in immunosuppressed mice were investigated . ICR mice, which were immunosuppressed by injection of cyclophosphamide or prednisolone, were given these medicines orally andchallenged intravenously with Candida albicans (day 0) . Treatments with a daily dose of 1 g/kg/day of TJ-48 or that of 1 or 2 g/kg/day of TJ-108 for 4 consecutive days from day-4 significantly prolonged the survival time of the Candida-infected mice pretreated with cyclophosphamide . Treatments with a daily dose of 1 g/kg/day of TJ-48 for 4 consecutive days from day 0, but not from day -4, significantly prolonged the life span of the Candida-infected mice pretreated with prednisolone . On the basis of these results and previous findings, characteristics of these kampo medicines as therapeutic agents against candidiasis in immunosuppressed hosts were discussed. Med Microbiol Immunol (Berl), 1999 Dec, 188(3), 117 - 24 Candidacidal activity of shortened synthetic analogs of amoebapores and NK-lysin; Andra J et al.; Natural antimicrobial peptides and synthetic analogs thereof have emerged as compounds with potentially significant therapeutical application against human pathogens . Amoebapores are 77-residue peptides with cytolytic and antibacterial activity considered to act by forming ion channels in cytoplasmic membranes of the victim cells . A functionally and structurally similar peptide named NK-lysin exists in mammalian lymphocytes . Several synthetic analogs of amoebapores and NK-lysin, which are substantially reduced in size compared to the parent molecules, were tested for their ability to inhibit the growth of and to kill Candida albicans . Some of the peptides displayed potent activity against a clinical isolate as well as against defined culture strains . Among the most active peptides found are some shortened substitution analogs of amoebapore C and a cationic core region of NK-lysin . As these peptides are also highly active against Gram-positive and Gram-negative bacteria but are of low cytotoxicity towards a human keratinocyte cell line they may provide promising templates for the design of broad-spectrum peptide antibiotics. J Hum Lact, 1999 Dec, 15(4), 281 - 8 Mammary candidosis in lactating women; Heinig MJ et al.; Though perceived to be a growing problem by lactation professionals, fungal infection of the breast (mammary candidosis) is largely unstudied . Candida albicans, a commensal organism encountered frequently in the vagina and gastrointestinal tract of humans, has been reported to be responsible for both superficial (cutaneous) and localized (ductal) infection of the mammary gland in lactating women, though the latter association is not universally accepted . Severe pain is considered to be characteristic of yeast infection of the breast and may be a cause of premature weaning among lactating mothers . Given that pain is often the complaint that prompts mothers to consult lactation professionals, it is important that healthcare providers working with lactating women be knowledgeable about this disease . In this article, current research regarding yeast infection of the breast is summarized, including morphology and pathology, diagnosis, risk factors, and common treatment options. J Invertebr Pathol, 2000 Feb, 75(2), 107 - 16 Comparative analysis of the binding of antibodies prepared against the insect Spodoptera exigua and against the mycopathogen Nomuraea rileyi; Pendland JC et al.; Polyclonal antibodies were produced in mice against Spodoptera exigua (beet armyworm) larval hemolymph and hemocytes and against cell wall surfaces of hyphal bodies and hyphae of the entomopathogenic hyphomycete Nomuraea rileyi . In addition to exhibiting strong activity against their original antigenic substrates, all of the antibodies cross-react extensively with other substrates . The hemolymph antibody binds to hemocytes and vice versa, and both antibodies cross-react to the insect fat body basement membrane (extracellular matrix (ECM) and to N . rileyi and Beauveria bassiana (another entomopathogenic fungus) cell wall surfaces (ECM) . Likewise, the anti-fungal antibodies cross-react with S . exigua hemolymph and hemocytes, especially the granules that may contain ECM components, and with fat body basement membrane . These cross-reactivities are specific as indicated by negative controls in the microscopy and Western blotting assays . Parallel labeling experiments using Con A suggest that the reactive epitopes contain mannose; however, none of the antibodies bind to mannose residues of nonentomopathogenic Candida albicans or Saccharomyces cerevisiae yeast cells . Thus, these cross-reactivities suggest that the host mimicry expressed by surface components of entomopathogenic fungi represents an important pathogenic determinant. Antimicrob Agents Chemother, 2000 May, 44(5), 1200 - 8 Enhanced extracellular production of aspartyl proteinase, a virulence factor, by Candida albicans isolates following growth in subinhibitory concentrations of fluconazole; Wu T et al.; We examined the production of secreted aspartyl proteinase (Sap), a putative virulence factor of Candida albicans, by a series of 17 isolates representing a single strain obtained from the oral cavity of an AIDS patient before and after the development of clinical and in vitro resistance to fluconazole . Isolates were grown in Sap-inducing yeast carbon base-bovine serum albumin medium containing 0, 0.25, 0.5, or 1 MIC of fluconazole, and cultures were sampled daily for 14 days to determine extracellular Sap activity by enzymatic degradation of bovine serum albumin . Extracellular Sap activity was significantly decreased in a dose-dependent manner for the most fluconazole-susceptible isolate (MIC, 1.0 microg/ml) and significantly increased in a dose-dependent manner for the most fluconazole-resistant isolate (MIC, >64 microg/ml) . Enhanced extracellular Sap production could not be attributed to cell death or nonspecific release of Sap, because there was no reduction in the number of CFU and no significant release of enolase, a constitutive enzyme of the glycolytic pathway . Conversely, intracellular Sap concentrations were significantly increased in a dose-dependent manner in the most fluconazole-susceptible isolate and decreased in the most fluconazole-resistant isolate . Enhanced Sap production correlated with the overexpression of a gene encoding a multidrug resistance (MDR1) efflux pump occurring in these isolates . These data indicate that exposure to subinhibitory concentrations of fluconazole can result in enhanced extracellular production of Sap by isolates with the capacity to overexpress MDR1 and imply that patients infected with these isolates and subsequently treated with suboptimal doses of fluconazole may experience enhanced C . albicans virulence in vivo. Infect Immun, 2000 May, 68(5), 2464 - 9 Candida albicans and Candida krusei differentially induce human blood mononuclear cell interleukin-12 and gamma interferon production; Xiong J et al.; Protection against Candida infection involves both innate and acquired immune responses, and cytokines produced by monocytes during the innate response may modify the acquired immune response by T cells . We hypothesized that Candida species which differ in pathogenicity can differentially induce production of immunoregulatory cytokines by human monocytes, which in turn modify T cells for immune responses to Candida . To test this hypothesis, we examined the effects of Candida albicans and Candida krusei on immunoregulatory cytokine production by human monocytes and gamma interferon (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) . Purified monocytes were incubated with live or heat-killed strains of C . albicans and C . krusei at the optimal Candida/monocyte ratio of 0.5 . Cytokines in the supernatants were measured by enzyme-linked immunosorbent assay . Our data demonstrated that live C . albicans and C . krusei significantly induced interleukin-10 (IL-10), monocyte chemotactic factor 1, IL-1beta, and tumor necrosis factor alpha production by monocytes relative to unstimulated monocytes . In contrast, unlike C . krusei, pathogenic live strains of C . albicans induced no or only a minimal level of IL-12 . The expression of IL-12 p40 mRNA levels by reverse transcription-PCR corroborated the IL-12 protein (p70) findings . In human PBMC, human blood monocytes were the major source of both IL-10 and IL-12 production in response to C . albicans and C . krusei . Upon activation of T cells in the presence of Candida-modified monocytes and antigen-presenting cells, IL-12 production by PBMC treated with Candida organisms correlated strongly with the level of IFN-gamma production by T cells . These results indicate that the virulence of C . albicans may be related to its ability to induce the monocytic type II cytokine IL-10, with a selective inhibition of IL-12 production, which may be responsible for the observed lack of T-cell IFN-gamma and may restrain an effective type I immune response to Candida. Electrophoresis, 2000 Mar, 21(5), 956 - 61 Cell surface proteins of Candida albicans: preparation of extracts and improved detection of proteins; Vediyappan G et al.; We have reexamined the detection of the components in a beta-mercaptoethanol and ammonium carbonate buffer extract of surface proteins of Candida albicans and the effects of postextraction manipulation of the extract on recovery of extract components . Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), preferential staining of some moieties was observed when bands detected by a commercial silver staining method or a Coomassie Brilliant Blue (CBB) staining method were compared . Additional protein bands that were either not detected or poorly detected by a single method alone were readily observed by a combined silver-CBB staining method . This method also detected alterations in the profile of extracted proteins from organisms grown in the presence of galactose or hemoglobin rather than glucose . Two-dimensional electrophoresis (2-DE) gel analysis by double stain showed better detection of several acidic and basic protein spots . Less than 10% of the extract as determined by a dye-binding assay was lost following either or both lyophilization and dialysis . These manipulations of the extract did not change the protein profile following SDS-PAGE as determined by the combined staining or Western blot analysis of a 70 kDa protein . These observations suggest that soluble cell wall proteins are not unusually sensitive to procedures routinely used in protein purification . In addition, these studies suggest that a modified staining method that combines both silver stain and CBB stain provides improved detection of cell wall proteins compared to either method alone. FEMS Immunol Med Microbiol, 2000 May, 28(1), 15 - 23 Membrane attack complex formation on yeast as trigger of selective release of terminal complement proteins from human polymorphonuclear leukocytes; Lukasser-Vogl E et al.; It has recently been shown that measurable amounts of complement proteins, C6 and in particular C7, are released from human polymorphonuclear leukocytes (PMNs) . The aim of the present study was to investigate the impact of opsonized Candida albicans on this release . Stimulation with opsonized C . albicans led to a rapid and sustained increase of C6 and C7 in the cell culture supernatant beginning within 5 min of placing in co-culture, whereas co-culture with unopsonized C . albicans or C . albicans mock-opsonized with inactivated human serum did not affect the release . In contrast, even after stimulation employing opsonized C . albicans, no release of the complement component C8 and only trace amounts of C9 were detected . The presence of the membrane attack complex (MAC) on C . albicans after opsonization was demonstrated by indirect immunofluorescence . Opsonization of C . albicans with human serum deficient in or depleted of a terminal complement component resulted in only minor stimulation of C6 and C7 release, although C3 deposition on the surface of C . albicans was not affected as determined by direct immunofluorescence . Detailed analyses with inactivated or deficient sera showed that detection of C6 and C7 was not due to insufficient washing of the opsonized yeast prior to co-culture and suggest that only a small proportion of these proteins was derived from the membrane bound and then cleaved off MAC . Thus, these findings imply that MAC on the fungal surface may represent an additional trigger for the release of C6 and C7 from PMNs, suggesting a new role for the terminal complement complex (TCC) on target membranes as modulator of PMN functions locally at the site of inflammation. Eur Spine J, 2000 Feb, 9(1), 72 - 4 Spondylodiscitis and epidural abscess due to Candida albicans; Derkinderen P et al.; A 32-year-old woman, addicted to heroin, presented with a dorsal spondylodiscitis due to Candida albicans associated with epidural abscess . Antimycotic treatment was successful, and no neurosurgical decompression was necessary . To our knowledge, this is the first case of documented epidural involvement in candidal spondylodiscitis . The diagnosis of candidal spondylodiscitis should be considered in cases of para- or tetraplegia occurring in intravenous drug abusers. Diagn Microbiol Infect Dis, 2000 Apr, 36(4), 215 - 23 A global evaluation of the susceptibility of Candida species to fluconazole by disk diffusion . Global Antifungal Surveillance Group; Meis J et al.; An improved fluconazole 25-mg disk diffusion method was used to test the susceptibility of 20,900 consecutively isolated clinical strains of Candida species from 40 hospital laboratories in 26 countries . The procedure is similar to the National Committee for Clinical Laboratory Standards (NCCLS) M2-A6 method for testing bacteria, except Mueller-Hinton agar is supplemented with 2% glucose and 0.5 mcg/mL methylene blue . Plates were incubated at 35 degrees C and read after 18 to 24h . Tentative zone interpretive criteria were based on the correlation by regression analysis with the NCCLS M27-A Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts: > =19mm Susceptible, < = 12mm Resistant, and 13-18 mm Susceptible-Dose Dependent . Of 14,368 isolates of Candida albicans, 2,073 C . glabrata, 869 C . tropicalis, 752 C . parapsilosis, and 351 C . krusei, 99%, 67%, 90%, 94%, and 26%, respectively, were susceptible in vitro to fluconazole . All study sites used the BIOMIC System to electronically read zones on plates, interpret, record test results and verify quality control data . This is the largest study to date that evaluated a broad range of sequentially collected yeasts from various infections and different types of hospitals . The modified disk testing procedure is facile and economical to perform and offers a reproducible and accurate means to assess the in vitro susceptibility of Candida species to fluconazole. J Infect Dis, 2000 Apr, 181(4), 1441 - 6 Epub 2000 Apr 13. Estrogen effects on Candida albicans: a potential virulence-regulating mechanism; Zhang X et al.; Three Candida albicans strains were tested in the presence of 17-beta-estradiol (10-6 M and 10-9 M) for increased growth and for enhanced survival during incubation at nonpermissive temperatures . All 3 test organisms showed increased growth in the presence of estradiol compared with estrogen-free controls . Likewise, all 3 strains, when treated with estradiol, survived incubation at 48 degrees C better than did controls . Cytoplasmic extracts were probed with an anti-hsp90 antibody, and results suggested that intracellular hsp90 was up-regulated in the presence of 10-9 M 17-beta-estradiol . The results were confirmed by reverse-transcriptase polymerase chain reaction with primers specific for C . albicans hsp90 . A kinetic study revealed that peak hsp90 expression occurred within 2 h of exposure to 17-beta-estradiol . In addition, estrogen increased the amount of cdr1 (Candida multidrug resistance) mRNA compared with cells not treated with estrogen . Coumarin and phenol also up-regulated hsp90 and cdr1 mRNAs, indicating that the estrogen-sensing and -response systems in C . albicans may lack specificity. J Infect, 2000 Jan, 40(1), 88 - 90 Case report of three Candida albicans infections detected at delivery; Arnavielhe S et al.; We report three similar cases of Candida albicans infections in neonates, at delivery . A retrospective study of the isolates was conducted to define the diversity of infective strains and their susceptibility to amphotericin B and fluconazole . Three neonates with fever, 'not doing well' at delivery had positive cultures for C . albicans . Samples were then taken from the mothers who did not exhibit any clinical symptoms of infection . Candida albicans strains isolated from both neonates and mothers were cultured, six colonies of each were typed by multilocus enzyme electrophoresis . The E-test method was used to determine the susceptibility of each colony to the two antifungals commonly used in this unit: amphotericin B and fluconazole . The initial isolates were composed of different types of strains . In the three cases, one of the mother types was found in the neonate isolates, leading us to suggest a vertical transmission of strains . All of the other types were distinct . All of the types were susceptible to amphotericin B, although three of them, one type isolated from a neonate and two types isolated from the mother, were resistant to fluconazole . The diversity of infective strains remains alarming and encourages the consideration of several colonies per isolate or several isolates, when it is possible, per infection case . This study also points out the need to survey the susceptibility of infective strains, since some of them appear soon to be resistant to fluconazole. Int J Tissue React, 1999, 21(4), 93 - 104 Resveratrol, a natural stilbene in grapes and wine, enhances intraphagocytosis in human promonocytes: a co-factor in antiinflammatory and anticancer chemopreventive activity; Bertelli AA et al.; Trans-resveratrol, a natural stilbene present in wine and grapes, has been studied mainly for its antiinflammatory and anticancer activities . In this study the activity of resveratrol on proliferative immunological parameters (differentiation, apoptosis, phagocytosis and intracellular killing) was studied using a U937 human promonocytic cell line in comparison with another polyphenol, quercetin . After incubation of the pathogen, Candida albicans, intracellular killing by macrophage-like cells was decreased by quercetin and resveratrol 10 microM but was enhanced by resveratrol 1 microM after 20 h of treatment . Phagocytosis rate, expressed as phagocytosis frequency, (i.e., percentage number of phagocytosing cells/total cells) at 20 h was highest with resveratrol 10 microM and was higher with quercetin 10 microM than with resveratrol 1 microM . The phagocytosis index exhibited the same trend . While both polyphenols demonstrated cytostatic activity on U937 growth, a prointraphagocytic effect for resveratrol 10 microM-treated cells at 10 min, resveratrol 1 microM-treated cells at 20 h and resveratrol 10 microM-treated cells at 48 h was observed . Morphological examination with optic microscopy demonstrated both apoptotic and differentiating cells, even after 10 min treatment . Resveratrol-induced apoptosis (following 4 h treatment) was confirmed by flow cytometry at concentrations as low as 1 microM and 100 nM in the assay for detection of membrane phosphatidylserine . Resveratrol- or quercetin-treated, but unstimulated cells, did not produce tumor necrosis factor-alpha protein . As phosphatidylserine externalization triggers specific recognition by monocytes and macrophages, removal of intact apoptotic cells is important a) in cell population selection and differentiation for antiblastic therapy, and b) in preventing the release of toxic inflammatory substances such as reactive oxygen substances and proteolytic enzymes by dying cells . This observation suggests that wine polyphenols, at the same concentrations as those found in plasma after moderate wine consumption, are important cofactors in antiinfective, antiinflammatory and anticancer nonspecific immune reactions. Crit Rev Oral Biol Med, 1999, 10(1), 99 - 116 Denture plaque and adherence of Candida albicans to denture-base materials in vivo and in vitro; Radford DR et al.; The aim of this paper is to review our understanding of the mechanisms and clinical significance of adhesion of C . albicans to denture-base materials in relation to denture plaque and denture-related stomatitis . Earlier reports in the literature of a 65% prevalence level of denture-related stomatitis seem to be exaggerated . More recent studies indicate that denture-related stomatitis is considerably less common, particularly in normal healthy subjects . The etiology of the condition is discussed in this review, and although much of the literature supports the view that the condition is strongly associated with C . albicans, this is not always so . In some subjects, the cause appears to be related to a non-specific plaque . This review also considers the role of denture plaque in the pathogenesis of denture-related stomatitis, the sequential development of denture plaque, and its colonization by Candida organisms . Designing controlled in vivo studies is difficult, and as a consequence, many investigators have had to resort to in vitro studies . The majority of these studies have attempted to investigate the hydrophobicity of C . albicans, relating the surface free-energy of denture-base materials, particularly acrylic resin, to that of the organism . Surprisingly little work has been directed at surface roughness and how it affects retention of organisms . Further, no attention has been paid to the properties and character of the surface, other than average surface roughness, as it affects adhesion . A comparison of results from in vitro studies on the effect on adhesion of pre-coating the surfaces of denture-base materials with saliva has produced equivocal conclusions . This is largely due to little standardization of experimental protocols between studies, particularly in the collection and handling of the saliva used . In conclusion, the review strongly supports the suggestion that adherence of C . albicans to denture-base materials in vitro is related to the hydrophobicity of the organism . The clinical significance of the observation and the mechanisms for the development and maturation of denture plaque are yet to be understood . There is a clear need for further investigation of other factors that may moderate the adhesion of organisms and subsequent colonization of dent