Antimicrob Agents Chemother, 2004 Sep, 48(9), 3407 - 11 Paradoxical effect of caspofungin: reduced activity against Candida albicans at high drug concentrations; Stevens DA et al.; Resistance problems with caspofungin, an echinocandin inhibitor of fungal cell wall glucan synthesis, have been rare . We noted paradoxical turbid growth of Candida albicans isolates in broth in some high (supra-MIC) concentrations . Among isolates submitted for susceptibility testing and screened at drug concentrations up to 12.5 microg/ml, the frequency was 16% . Analysis of the turbid growth indicated slowing of growth in the presence of drug but with numbers of CFU up to 72% those of drug-free controls . Clearing of growth again by the highest drug concentrations produced a quadriphasic pattern in a tube dilution series . Cells growing at high drug concentrations were not resistant on retesting but showed the paradoxical effect of the parent . Among a selected series of isolates tested at concentrations up to 50 microg/ml, an additional 53% showed a "mini-paradoxical effect": no turbid growth but incomplete killing at high concentrations (supra-minimum fungicidal concentration) . These effects were reproducible; medium dependent in extent; noted in macro- and microdilution, in the presence or absence of serum, and on agar containing drug (but not when drug concentrations were not constant, as in agar diffusion); not seen with other echinocandins and less commonly in other Candida species; and not due to destruction of drug in tubes showing the effect . Cooperative enhancement of inhibition by a second drug could eradicate the effect . We postulate that high drug concentrations derepress or activate resistance mechanisms . The abilities of subpopulations to survive at high drug concentrations could have in vivo consequences. Antimicrob Agents Chemother, 2004 Sep, 48(9), 3272 - 8 The novel azole R126638 is a selective inhibitor of ergosterol synthesis in Candida albicans, Trichophyton spp., and Microsporum canis; Vanden Bossche H et al.; R126638 is a novel triazole with in vitro activity similar to that of itraconazole against dermatophytes, Candida spp., and Malassezia spp . In animal models of dermatophyte infections, R126638 showed superior antifungal activity . R126638 inhibits ergosterol synthesis in Candida albicans, Trichophyton mentagrophytes, Trichophyton rubrum, and Microsporum canis at nanomolar concentrations, with 50% inhibitory concentrations (IC(50)s) similar to those of itraconazole . The decreased synthesis of ergosterol and the concomitant accumulation of 14 alpha-methylsterols provide indirect evidence that R126638 inhibits the activity of CYP51 that catalyzes the oxidative removal of the 14 alpha-methyl group of lanosterol or eburicol . The IC(50)s for cholesterol synthesis from acetate in human hepatoma cells were 1.4 microM for itraconazole and 3.1 microM for R126638 . Compared to itraconazole (IC(50) = 3.5 microM), R126638 is a poor inhibitor of the 1 alpha-hydroxylation of 25-hydroxyvitamin D(3) (IC(50) > 10 microM) . Micromolar concentrations of R126638 and itraconazole inhibited the 24-hydroxylation of 25-hydroxyvitamin D(3) and the conversion of 1,25-dihydroxyvitamin D(3) into polar metabolites . At concentrations up to 10 microM, R126638 had almost no effect on cholesterol side chain cleavage (CYP11A1), 11 beta-hydroxylase (CYP11B1), 17-hydroxylase and 17,20-lyase (CYP17), aromatase (CYP19), or 4-hydroxylation of all-trans retinoic acid (CYP26) . At 10 microM, R126638 did not show clear inhibition of CYP1A2, CYP2A6, CYP2D6, CYP2C8, CYP2C9, CYP2C10, CYP2C19, or CYP2E1 . Compared to itraconazole, R126638 had a lower interaction potential with testosterone 6 beta hydroxylation and cyclosporine hydroxylation, both of which are catalyzed by CYP3A4, whereas both antifungals inhibited the CYP3A4-catalyzed hydroxylation of midazolam similarly . The results suggest that R126638 has promising properties and merits further in vivo investigations for the treatment of dermatophyte and yeast infections. Oral Microbiol Immunol, 2004 Oct, 19(5), 293 - 6 Differential invasion of Candida albicans isolates in an in vitro model of oral candidosis; Bartie KL et al.; The study assessed the ability of Candida albicans isolates to invade an in vitro oral tissue model . The extent and pattern of isolate invasion was then correlated with the infection origin of the isolate to identify characteristics that may be restricted to specific forms of oral infection, particularly chronic hyperplastic candidosis (CHC) . Reconstituted human oral epithelium was infected with C . albicans isolated from normal oral mucosa (n = 4), CHC (n = 7), non-CHC oral candidoses (n = 4) and squamous cell carcinoma (SCC; n = 4) . After infection for 24 h, histological analysis revealed yeast adhesion, hyphal extension, and invasion of the epithelium . Differential patterns of invasion were evident and, whilst consistent for a given isolate, did not relate to the infection origin of the isolate . Two principal patterns of invasion were evident and described as either a 'localised' or a 'uniform' distribution of invading hyphae . Several isolates also exhibited superficial infection with limited hyphal invasion . In conclusion, the use of the in vitro tissue model allowed the assessment of the invasive capabilities of isolates of C . albicans . However, the apparent differences in invasive characteristics did not appear to be related to the clinical origin of isolates. Med Pregl, 2004 Jan-Feb, 57(1-2), 30 - 2 {The role of parasites and fungi in secondary infertility}; Candida albicans osteomyelitis: case report and literature review; Servicio de Enfermedades Infecciosas del Adulto, Hospital Universitario de Caracas, VenezuelaOBJECTIVE: Osteomyelitis due to Candida species is an unusual but recognized entity . However, with the increasing occurrence of factors predisposing to candidemia and invasive candidiasis, candidal osteomyelitis is being diagnosed more frequently . An unusual case of Candida albicans osteomyelitis is reported here, along with a review of the published literature on previously reported cases of this disease . METHODS: Report of the case and literature review . RESULTS: In this case, Candida albicans was isolated from the talus; however, the diagnosis was made after several cultures were performed . Only one other case of Candida osteomyelitis located in foot bones was found in the review . CONCLUSIONS: It is thought that this is the first case reported in Venezuela, and only the second in international literature. Biochem Biophys Res Commun, 2004 Sep 17, 322(2), 520 - 5 Disulfiram is a potent modulator of multidrug transporter Cdr1p of Candida albicans; Shukla S et al.; To find novel drugs for effective antifungal therapy in candidiasis, we examined disulfiram, a drug used for the treatment of alcoholism, for its role as a potential modulator of Candida multidrug transporter Cdr1p . We show that disulfiram inhibits the oligomycin-sensitive ATPase activity of Cdr1p and 2.5mM dithiothreitol reverses this inhibition . Disulfiram inhibited the binding of photoaffinity analogs of both ATP ({alpha-(32)P}8-azidoATP; IC(50)=0.76 microM) and drug-substrates ({(3)H}azidopine and {(125)I}iodoarylazidoprazosin; IC(50) approximately 12 microM) to Cdr1p in a concentration-dependent manner, suggesting that it can interact with both ATP and substrate-binding site(s) of Cdr1p . Furthermore, a non-toxic concentration of disulfiram (1 microM) increased the sensitivity of Cdr1p expressing Saccharomyces cerevisiae cells to antifungal agents (fluconazole, miconazole, nystatin, and cycloheximide) . Collectively these results demonstrate that disulfiram reverses Cdr1p-mediated drug resistance by interaction with both ATP and substrate-binding sites of the transporter and may be useful for antifungal therapy. Infect Immun, 2004 Sep, 72(9), 4948 - 55 Global cell surface conformational shift mediated by a Candida albicans adhesin; Rauceo JM et al.; Candida albicans maintains both commensal and pathogenic states in humans . Both states are dependent on cell surface-expressed adhesins, including those of the Als family . Heterologous expression of Als5p at the surface of Saccharomyces cerevisiae results in Als5p-mediated adhesion to various ligands, followed by formation of multicellular aggregates . Following adhesion of one region of the cell to fibronectin-coated beads, the entire surface of the cells became competent to mediate cell-cell aggregation . Aggregates formed in the presence of metabolic inhibitors or signal transduction inhibitors but were reduced in the presence of 8-anilino-1-naphthalene-sulfonic acid (ANS) or Congo Red (CR), perturbants that inhibit protein structural transitions . These perturbants also inhibited aggregation of C . albicans . An increase in ANS fluorescence, which accompanied Als-dependent cellular adhesion, indicated an increase in cell surface hydrophobicity . In addition, C . albicans and Als5p-expressing S . cerevisiae showed an aggregation-induced birefringence indicative of order on the cell surface . The increase in birefringence did not occur in the presence of the aggregation disruptants ANS and CR . These results suggest a model for Als5p-mediated aggregation in which an adhesion-triggered change in the conformation of Als5p propagates around the cell surface, forming ordered aggregation-competent regions. Infez Med, 2004 Jun, 12(2), 136 - 8 Late onset opportunistic infections in a renal allograft recipient: a case report; Morosi S et al.; In renal allograft recipients, infection disease complications remain an important cause of morbidity and mortality during the post-transplant period . This complication occurs more frequently from 1 to 6 months after transplant . The epidemiology of infection during the postoperative period is less well characterized, because recipients routinely reside at home . We describe a case of late onset Candida albicans and HSV-1 esophagitis, and Pneumocystis carinii pneumonia, that occurs 9 years after renal transplantation in a patient with severe CD4+ T-lymphocytopenia and hypogammaglobulinaemia . We underline the importance of monitoring immunosuppressive therapy in these patients and the usefulness of prophylaxis against P . carinii pneumonia J Lipid Res, 2004 Nov, 45(11), 2000 - 7 Epub 2004 Aug 16. Fluconazole binding and sterol demethylation in three CYP51 isoforms indicate differences in active site topology; Bellamine A et al.; 14alpha-Demethylase (CYP51) is a key enzyme in all sterol biosynthetic pathways (animals, fungi, plants, protists, and some bacteria), catalyzing the removal of the C-14 methyl group following cyclization of squalene . Based on mutations found in CYP51 genes from Candida albicans azole-resistant isolates obtained after fluconazole treatment of fungal infections, and using site-directed mutagenesis, we have found that fluconazole binding and substrate metabolism vary among three different CYP51 isoforms: human, fungal, and mycobacterial . In C . albicans, the Y132H mutant from isolates shows no effect on fluconazole binding, whereas the F145L mutant results in a 5-fold increase in its IC(50) for fluconazole, suggesting that F145 (conserved only in fungal 14alpha-demethylases) interacts with this azole . In C . albicans, F145L accounts, in part, for the difference in fluconazole sensitivity reported between mammals and fungi, providing a basis for treatment of fungal infections . The C . albicans Y132H and human Y145H CYP51 mutants show essentially no effect on substrate metabolism, but the Mycobacterium tuberculosis F89H CYP51 mutant loses both its substrate binding and metabolism . Because these three residues align in the three isoforms, the results indicate that their active sites contain important structural differences, and further emphasize that fluconazole and substrate binding are uncoupled properties. Biochemistry, 2004 Aug 24, 43(33), 10789 - 99 CYP51 from Trypanosoma brucei is obtusifoliol-specific; Lepesheva GI et al.; New isoforms of CYP51 (sterol 14alpha-demethylase), an essential enzyme in sterol biosynthesis and primary target of azole antimycotic drugs, are found in pathogenic protists, Trypanosoma brucei(TB), T . vivax, T . cruzi, and Leishmania major . The sequences share approximately 80% amino acid identity and are approximately 25% identical to sterol 14alpha-demethylases from other biological kingdoms . Differences of residues conserved throughout the rest of the CYP51 family that align with the BC-loop and helices F and G of CYP51 from Mycobacterium tuberculosis (MT)) imply possible alterations in the topology of the active site cavity of the protozoan enzymes . CYP51 and cytochrome P450 reductase (CPR) from TB were cloned, expressed in Escherichia coli, and purified . The P450 has normal spectral features (including absolute absorbance, carbon monoxide, and ligand binding spectra), is efficiently reduced by TB and rat CPR but demonstrates altered specificity in comparison with human CYP51 toward three tested azole inhibitors, and contrary to the human, Candida albicans, and MT isoforms, reveals profound substrate preference toward obtusifoliol (turnover 5.6 min(-1)) . It weakly interacts with the other known CYP51 substrates; slow lanosterol conversion predominantly produces the 14alpha-carboxyaldehyde intermediate . Although obtusifoliol specificity is typical for plant isoforms of CYP51, the set of sterol biosynthetic enzymes in the protozoan genomes together with available information about sterol composition of kinetoplastid cells suggest that the substrate preference of TBCYP51 may reflect a novel sterol biosynthetic pathway in Trypanosomatidae. Mycoses, 2004 Aug, 47(7), 330 - 7 Influence of voriconazole and fluconazole on reconstituted multilayered oesophageal epithelium infected by Candida albicans; Bernhardt J et al.; Reconstituted multilayered oesophageal epithelium appears to be a good basis to test the efficacy of voriconazole (VOR) and fluconazole (FLU) in the tissue . The resulting model of a Candida oesophagitis was approaching the in vivo situation . We infected the tissue with 2 x 10(6) cfu of the Candida albicans strain SC5314 . In the trials with FLU we also used clinical strains . Four hours after infection a good growth of C . albicans appeared mainly with hyphae on the surface of the tissue and a tendency to invasion . The destruction of the tissue began after 36 h . VOR (2 and 16 microg ml-1, respectively) prevented the penetration of hyphae into the tissue, when it was given 4-8 h after infection . It was less effective in reduction of Candida growth on the tissue surface . When VOR was given 16-24 h postinfection, the Candida infiltration stopped more slowly . Thirty-six hours after infection VOR application could not stop the destruction of the tissue despite reducing the fungi . The results with FLU (32 microg ml-1) were in principle the same, but not so distinct . FLU seems to be more effective against clinical strains of C . albicans than against the type strain. Mycoses, 2004 Aug, 47(7), 304 - 9 Estrogen-dependent induction of persistent vaginal candidosis in naïve mice; Hamad M et al.; The capacity of estrogen to induce vaginal candidosis (VC) in the absence of previous or concurrent Candida albicans infections was examined . Adult female Balb/c mice were evaluated for vaginal C . albicans burden, C . albicans-specific delayed type hypersensitivity (DTH) responses and vaginal lymphocyte population kinetics at several time-points while receiving weekly injections of 0.5 mg estrogen . Estrogen treatment resulted in the appearance of significant levels of C . albicans vaginal colonization, which persisted for a period of 4 weeks . A marked suppression of DTH responses mounted against subsequent C . albicans challenge was observed . Absolute number of vaginal T lymphocytes gradually increased by several folds especially at weeks 5-6 following the start of estrogen treatment . These results clearly indicate that estrogen, independent of other predisposing factors, is capable of perturbing the commensal relationship between the host and the fungus, which results in the induction of persistent VC. Mycoses, 2004 Aug, 47(7), 300 - 3 Yeasts species distribution in Neonatal Intensive Care Units in northeast Argentina; Giusiano GE et al.; The aim of this study was to determine the distribution and antifungal susceptibility profile of yeast species isolated from neonates in Neonatal Intensive Care Units (NICU) in northeast of Argentina . With this purpose 92 strains isolated from 25 blood stream cultures, 20 venous catheters, 23 suprapubic aspirations and 24 rectal swabs were studied . Candida albicans and C . parapsilosis appeared with similar frequencies (36%) in blood stream isolates . Candida parapsilosis (50%) was the most frequent catheters colonizer and C . tropicalis (54.2%) was the most frequent yeast associated with gastrointestinal tract colonization . Candida krusei, C . glabrata and Trichosporon cutaneum appeared with a very low frequency . A high rate of susceptibility to amphotericin B, fluconazole, and itraconazole was observed. Mol Microbiol, 2004 Aug, 53(4), 1209 - 20 A family of Candida cell surface haem-binding proteins involved in haemin and haemoglobin-iron utilization; Weissman Z et al.; The ability to acquire iron from host tissues is a major virulence factor of pathogenic microorganisms . Candida albicans is an important fungal pathogen, responsible for an increasing proportion of systemic infections . C . albicans, like many pathogenic bacteria, is able to utilize haemin and haemoglobin as iron sources . However, the molecular basis of this pathway in pathogenic fungi is unknown . Here, we identify a conserved family of plasma membrane-anchored proteins as haem-binding proteins that are involved in haem-iron utilization . We isolated RBT51 as a gene that is sufficient by itself to confer to S . cerevisiae the ability to utilize haemoglobin iron . RBT51 is highly homologous to RBT5, which was previously identified as a gene negatively regulated by the transcriptional suppressor CaTup1 . Rbt5 and Rbt51 are mannosylated proteins that carry the conserved CFEM domain . We find that RBT5 is strongly induced by starvation for iron, and that deletion of RBT5 is by itself sufficient to significantly reduce the ability of C . albicans to utilize haemin and haemoglobin as iron sources . Iron starvation-inducible, antigenically cross-reacting haem-binding proteins are also present in other Candida species that are able to utilize haem-iron, underscoring the conservation of this iron acquisition pathway among pathogenic fungi. Mol Microbiol, 2004 Aug, 53(4), 1177 - 94 Homologous recombination in Candida albicans: role of CaRad52p in DNA repair, integration of linear DNA fragments and telomere length; Ciudad T et al.; Chromosomal rearrangements are common in both clinical isolates and spontaneous mutants of Candida albicans . It appears that many of these rearrangements are caused by translocations around the major sequence repeat (MSR) that is present in all chromosomes except chromosome 3, suggesting that homologous recombination (HR) may play an important role in the survival of this organism . In order to gain information on these processes, we have cloned the homologue of RAD52, which in Saccharomyces cerevisiae is the only gene required for all HR events . CaRAD52 complemented poorly a rad52 mutant of S . cerevisiae . Two null Carad52Delta/Carad52Delta mutants were constructed by sequential deletion of both alleles and two reconstituted strains were obtained by reintegration of the gene . Characterization of these mutants indicated that HR plays an essential role in the repair of DNA lesions caused by both UV light and the radiomimetic compound methyl-methane-sulphonate (MMS), whereas the non-homologous end-joining pathway (NHEJ) is used only in the absence of Rad52p or after extensive DNA damage . Repair by HR is more efficient in exponentially growing than in stationary cells, probably because a larger number of cells are in late S or G2 phases of the cell cycle (and therefore, can use a sister chromatid as a substrate for recombinational repair), whereas stationary phase cells are mainly in G0 or G1, and only can be repaired using the chromosomal homologue . In addition, CaRad52p is absolutely required for the integration of linear DNA with long flanking homologous sequences . Finally, the absence of CaRad52p results in the lengthening of telomeres, even in the presence of an active telomerase, an observation not described in any other organism . This raises the possibility that both telomerase and homologous recombination may function simultaneously at C . albicans telomeres. Parassitologia, 2004 Jun, 46(1-2), 89 - 93 {Highly Active AntiRetroviral Therapy and opportunistic protozoan infections}; Pozio E; Opportunistic parasite infections (OPIs) are an important cause of morbidity and mortality in persons infected with HIV . In industrialised countries, the use of Highly Active AntiRetroviral Therapy (HAART) results to be effective in suppressing the HIV viral load, with a quantitative and qualitative improvement in the CD4+ T-cell count followed by a strong reduction of opportunistic infections including those caused by parasites . These successes have been mainly attributed to the reconstitution of the cell immunity, which play the most important role in controlling OPIs . However, there are many clinical reports and several laboratory results, which suggest that the control of OPIs in HIV-positive persons under HAART is also induced by the anti-HIV protease inhibitors (PIs), which inhibit the aspartyl proteases of the parasites . The non-conventional use of HIV-PIs seems to be an alternative way for the treatment of parasitic infections, which should be deeply investigated . Of five longitudinal studies carried out before and after the introduction of HAART, four studies showed a strong reduction of toxoplasmic encephalitis (TE) in HIV-positive persons under HAART, whereas in another study, no difference was observed in the incidence rate of TE before and after the introduction of HAART . The influence of HAART in reducing TE has been also confirmed in a randomised, controlled clinical trial, which showed that there is no increase in the risk of developing TE after beginning HAART, even though HIV-infected persons with TE had a discontinuing prophylaxis for Toxoplasma gondii . Four HIV protease inhibitors were tested against the T . gondii virulent RH strain in vitro, alone or in association with pyrimethamine or sulfadiazine . Ritonavir and nelfinavir were highly inhibitory for the parasite growth . Furthermore, none of the antiviral drugs negatively affected the anti-Toxoplasma activity of pyrimethamine or sulfadiazine . In HIV-Leishmania co-infections, a changing pattern has been observed in the HAART era, characterised by a high rate of relapses, which could be explained by the increased survival rate resulting from the effective antiretroviral therapy . A 64.8% decrease of the visceral leishmaniasis incidence was detected after HAART began to be used extensively in Spain . In a large cohort study carried out in ten European countries and in Australia, the relative risk to contract cryptosporidiosis as the first AIDS defining disease was reduced by 96% in the HAART era . In Italy, the relative risk of death for cryptosporidiosis reduced of 74% in the period 1997-98, when HIV-positive persons received HAART . In a large study carried out in Italy, isosporiasis was included in the group of opportunistic infections, of which the relative hazards showed a reduction of 95% in the HAART era . Since 1997, there was the evidence that the use of HAART in persons with advanced HIV infection can improve chronic diarrhoea and lead to disappearance of Enterocytozoon bieneusi from the stools . Although the reconstitution of the cellular immunity seems to be the main factor influencing the reduction of OPIs in persons with AIDS who undergo HAART, there are clinical and microbiological evidences, as well as in vitro and in vivo results, which indicate direct effects of HIV-PIs on the proteases of opportunistic parasites . These findings stress the existence of non-conventional unexpected benefits of PIs in HAART against protozoa . In addition, this benefit of PIs has been demonstrated also for Candida albicans secreted aspartyl proteases and for P . carinii acid proteases . In spite of these important results, HIV PIs are still very toxic for humans, specially in cases of very long treatment, and no clinical trial has been carried out for persons at risk, such as children and pregnant women, because the priority was to reduce the severity of HIV and not the evaluation of possible side effects of the therapy . It follows that further researches are needed to establish the non-conventional use of HIV PIs . Furthermore, the study of PIs against specific aspartyl proteases of those opportunistic protozoa that cause severe and intractable diseases, could be considered an alternative way towards the development of new drugs that may prove effective against these infections. Phytother Res, 2004 Jul, 18(7), 542 - 5 Antifungal constituents of Melicope borbonica; Simonsen HT et al.; Fractionation of extracts of the leaves of Melicope borbonica (syn . Euodia borbonica var . borbonica), a medicinal plant from the Reunion Island that is traditionally used for wound healing and other ailments, afforded an acetophenone (xanthoxylin) and two coumarins, scoparone and limettin, as the major constituents . All three compounds exhibited moderate antifungal activity against Candida albicans and Penicillium expansum, in accordance with the traditional use of the plant . Moreover, 2,4,6-trimethoxyacetophenone (methylxanthoxylin), three other coumarins {7-(3-methyl-2-butenyloxy)-6-methoxycoumarin, cedrelopsin and psoralen}, two sesquiterpenes (alpha-curcumene and 3,6-epidioxy-1,10-bisaboladiene), eugenol, methyleugenol and a lignan (sesamin) were isolated . None of the isolated constituents exhibited antiin fl ammatory activity in vitro . No alkaloids were detected. Biol Pharm Bull, 2004 Aug, 27(8), 1281 - 4 Role of Ca2+/calmodulin signaling pathway on morphological development of Candida albicans; Sato T et al.; A human fungal pathogen, Candida albicans, varies from the yeast form to the hyphal form due to various external signals . This morphogenetic transformation has been implicated in the development of pathogenicity . In this report, we show that calmodulin inhibitors (TFP and W-7) and an adenylatecyclase inhibitor (MDL-12-330A) suppressed the hyphae formation of C . albicans . Furthermore, the expression of hyphae-specific mRNAs located downstream from the RAS1-cAMP pathway was inhibited by these inhibitors . Suppression of hyphae formation by TFP or W-7 was not inhibited by the addition of cAMP, and these inhibitors did not affect the amount of cAMP in C . albicans . These results suggest that the Ca2+/calmodulin pathway contributes to hyphae formation and is related to the RAS1-cAMP pathway. Yao Xue Xue Bao, 2004 Apr, 39(4), 292 - 5 {Therapeutic efficiency of amphotericin B liposome modified by RMP-7 to transport drug across blood brain barrier}; Zhang XB et al.; AIM: To study the therapeutic efficiency of amphotericin B liposome (AmB-L) targeting to the brain in mice with meningitis . METHODS: Amphotericin B liposome targeting to the brain were prepared by film-sonication method . Their concentration and encapsulation percentage were determined . The Candida albicans was injected into the brain of BALB/c mice and the meningitis model was set up . Then the therapeutic efficiency of amphotericin B liposome targeting to the brain was studied . RESULTS: The encapsulation percentage of amphotericin B liposome was 93.3% . The meningitis model was set up after the Candida albicans was injected into the brain of BALB/c mice for 2 h . The therapeutic efficiency was increased after conjugating RMP-7 (the commercial nama is Cereport) to the surface of amphotericin B liposome . CONCLUSION: The therapeutic efficiency of Amphotericin B liposome targeting to the brain in the mice with meningitis was better than that of the common amphotericin B liposome and the life of the mice in AmB-L-PEG-RMP-7 group was longer than that of the mice in AmB-L-PEG group and AmB-L-PEG + RMP-7 group. Eukaryot Cell, 2004 Aug, 3(4), 1015 - 27 The closely related species Candida albicans and Candida dubliniensis can mate; Pujol C et al.; Because Candida dubliniensis is closely related to Candida albicans, we tested whether it underwent white-opaque switching and mating and whether white-opaque switching depended on MTL homozygosity and mating depended on switching, as they do in C . albicans . We also tested whether C . dubliniensis could mate with C . albicans . Sequencing revealed that the MTLalpha locus of C . dubliniensis was highly similar to that of C . albicans . Hybridization with the MTLa1, MTLa2, MTLalpha1, and MTLalpha2 open reading frames of C . albicans further revealed that, as in C . albicans, natural strains of C . dubliniensis exist as a/alpha, a/a, and alpha/alpha, but the proportion of MTL homozygotes is 33%, 10 times the frequency of natural C . albicans strains . C . dubliniensis underwent white-opaque switching, and, as in C . albicans, the switching was dependent on MTL homozygosis . C . dubliniensis a/a and alpha/alpha cells also mated, and, as in C . albicans, mating was dependent on a switch from white to opaque . However, white-opaque switching occurred at unusually high frequencies, opaque cell growth was frequently aberrant, and white-opaque switching in many strains was camouflaged by an additional switching system . Mating of C . dubliniensis was far less frequent in suspension cultures, due to the absence of mating-dependent clumping . Mating did occur, however, at higher frequencies on agar or on the skin of newborn mice . The increases in MTL homozygosity, the increase in switching frequencies, the decrease in the quality of switching, and the decrease in mating efficiency all reflected a general deterioration in the regulation of developmental processes, very probably due to the very high frequency of recombination and genomic reorganization characteristic of C . dubliniensis . Finally, interspecies mating readily occurred between opaque C . dubliniensis and C . albicans strains of opposite mating type in suspension, on agar, and on mouse skin . Remarkably, the efficiency of interspecies mating was higher than intraspecies C . dubliniensis mating, and interspecies karyogamy occurred readily with apparently the same sequence of nuclear migration, fusion, and division steps observed during intraspecies C . albicans and C . dubliniensis mating and Saccharomyces cerevisiae mating. Eukaryot Cell, 2004 Aug, 3(4), 932 - 43 Discovery of Cercosporamide, a known antifungal natural product, as a selective Pkc1 kinase inhibitor through high-throughput screening; Sussman A et al.; The Pkc1-mediated cell wall integrity-signaling pathway is highly conserved in fungi and is essential for fungal growth . We thus explored the potential of targeting the Pkc1 protein kinase for developing broad-spectrum fungicidal antifungal drugs through a Candida albicans Pkc1-based high-throughput screening . We discovered that cercosporamide, a broad-spectrum natural antifungal compound, but previously with an unknown mode of action, is actually a selective and highly potent fungal Pkc1 kinase inhibitor . This finding provides a molecular explanation for previous observations in which Saccharomyces cerevisiae cell wall mutants were found to be highly sensitive to cercosporamide . Indeed, S . cerevisiae mutant cells with reduced Pkc1 kinase activity become hypersensitive to cercosporamide, and this sensitivity can be suppressed under high-osmotic growth conditions . Together, the results demonstrate that cercosporamide acts selectively on Pkc1 kinase and, thus, they provide a molecular mechanism for its antifungal activity . Furthermore, cercosporamide and a beta-1,3-glucan synthase inhibitor echinocandin analog, by targeting two different key components of the cell wall biosynthesis pathway, are highly synergistic in their antifungal activities . The synergistic antifungal activity between Pkc1 kinase and beta-1,3-glucan synthase inhibitors points to a potential highly effective combination therapy to treat fungal infections. Eukaryot Cell, 2004 Aug, 3(4), 919 - 31 Gpr1, a putative G-protein-coupled receptor, regulates morphogenesis and hypha formation in the pathogenic fungus Candida albicans; Miwa T et al.; In response to various extracellular signals, the morphology of the human fungal pathogen Candida albicans switches from yeast to hypha form . Here, we report that GPR1 encoding a putative G-protein-coupled receptor and GPA2 encoding a Galpha subunit are required for hypha formation and morphogenesis in C . albicans . Mutants lacking Gpr1 (gpr1/gpr1) or Gpa2 (gpa2/gpa2) are defective in hypha formation and morphogenesis on solid hypha-inducing media . These phenotypic defects in solid cultures are suppressed by exogenously added dibutyryl-cyclic AMP (dibutyryl-cAMP) . Biochemical studies also reveal that GPR1 and GPA2 are required for a glucose-dependent increase in cellular cAMP . An epistasis analysis indicates that Gpr1 functions upstream of Gpa2 in the same signaling pathway, and a two-hybrid assay reveals that the carboxyl-terminal tail of Gpr1 interacts with Gpa2 . Moreover, expression levels of HWP1 and ECE1, which are cAMP-dependent hypha-specific genes, are reduced in both mutant strains . These findings support a model that Gpr1, as well as Gpa2, regulates hypha formation and morphogenesis in a cAMP-dependent manner . In contrast, GPR1 and GPA2 are not required for hypha formation in liquid fetal bovine serum (FBS) medium . Furthermore, the gpr1 and the gpa2 mutant strains are fully virulent in a mouse infection . These findings suggest that Gpr1 and Gpa2 are involved in the glucose-sensing machinery that regulates morphogenesis and hypha formation in solid media via a cAMP-dependent mechanism, but they are not required for hypha formation in liquid medium or during invasive candidiasis. Eukaryot Cell, 2004 Aug, 3(4), 900 - 9 Ectopic Expression of URA3 can influence the virulence phenotypes and proteome of Candida albicans but can be overcome by targeted reintegration of URA3 at the RPS10 locus; Brand A et al.; Uridine auxotrophy, based on disruption of both URA3 alleles in diploid Candida albicans strain SC5314, has been widely used to select gene deletion mutants created in this fungus by "Ura-blasting" and PCR-mediated disruption . We compared wild-type URA3 expression with levels in mutant strains where URA3 was positioned either within deleted genes or at the highly expressed RPS10 locus . URA3 expression levels differed significantly and correlated with the specific activity of Ura3p, orotidine 5'-monophosphate decarboxylase . Reduced URA3 expression following integration at the GCN4 locus was associated with an attenuation of virulence . Furthermore, a comparison of the SC5314 (URA3) and CAI-4 (ura3) proteomes revealed that inactivation of URA3 caused significant changes in the levels of 14 other proteins . The protein levels of all except one were partially or fully restored by the reintegration of a single copy of URA3 at the RPS10 locus . Transcript levels of genes expressed ectopically at this locus in reconstituted heterozygous mutants also matched the levels found when the genes were expressed at their native loci . Therefore, phenotypic changes in C . albicans can be associated with the selectable marker rather than the target gene . Reintegration of URA3 at an appropriate expression locus such as RPS10 can offset most problems related to the phenotypic changes associated with gene knockout methodologies. Yeast, 2004 Jul 30, 21(10), 883 - 94 Characterization of Candida albicans orthologue of the Saccharomyces cerevisiae signal-peptidase-subunit encoding gene SPC3; De La Rosa JM et al.; The Candida albicans orthologue of the SPC3 gene, which encodes one of the subunits essential for the activity of the signal peptidase complex in Saccharomyces cerevisiae, was isolated by complementation of a thermosensitive mutation in the S . cerevisiae SEC61 gene . The cloned gene (CaSPC3) encodes a putative protein of 192 amino acids that contains one potential membrane-spanning region and shares significant homology with the corresponding products from mammalian (Spc22/23p) and yeast (Spc3p) cells . CaSPC3 is essential for cell viability, since a hemizygous strain containing a single copy of CaSPC3 under control of the methionine-repressible MET3 promoter did not grow in the presence of methionine and cysteine . The cloned gene could rescue the phenotype associated with a spc3 mutation in S . cerevisiae, indicating that it is the true C . albicans orthologue of SPC3 . However, in contrast with results previously described for its S . cerevisiae orthologue, CaSPC3 was not able to complement the thermosensitive growth associated with a mutation in the SEC11 gene . The heterologous complementation of the sec61 mutant suggests that Spc3p could play a role in the interaction that it is known to occur between the translocon (Sec61 complex) and the signal peptidase complex, at the endoplasmic reticulum membrane. Int J Dev Biol, 2004 Jun, 48(4), 339 - 42 Developing chick embryos express a protein which shares homology with the nuclear pore complex protein Nup88 present in human tumors; Schneider J et al.; Nup88 is a nuclear pore complex protein which is overexpressed in a variety of human tumors of the stomach, colon, liver, pancreas, breast, lung, ovary, uterus, prostate and kidney . A monoclonal antibody crossreacting with the yeast Candida albicans and Nup88 was used to investigate the expression of cross-reactive antigens in chick embryos, in an attempt to identify an experimental model for studying the role played by Nup88 during cell development and differentiation . All cells in the trilaminar embryo were labeled with the antibody, but as development advanced and organogenesis was completed, expression of the corresponding antigen became more restricted . Thus, some structures continued to be intensely labeled (skin epithelium, oropharyngeal endothelium, perichondral mesenchymal tissue), whereas others ( muscular tissue, vascular endothelium, respiratory endothelium, digestive tract mucosa, peripheral nerves, medullary white matter and the retinal axons) were more moderately stained . No immunoreactivity was observed in the medullary grey matter or cartilage . A specific band of 53 kDa observed by Western blotting of chick embryo extracts suggested that the chicken antigen recognized by the monoclonal antibody is the homologue of human Nup88, which is associated with the high proliferation and low differentiation of tumor cells . The present results indicate that the role of Nup88 in cell differentiation and organ development could be fruitfully investigated using the developing chick embryo as an experimental model. Eur J Clin Microbiol Infect Dis, 2004 Sep, 23(9), 729 - 31 Epub 2004 Aug 06. Effects of caspofungin (MK-0991) and anidulafungin (LY303366) on phagocytosis, oxidative burst and killing of Candida albicans by human phagocytes; Frank U et al.; The objective of the present study was to investigate the influence of the new echinocandins caspofungin (MK-0991) and anidulafungin (LY303366) on human phagocytes . Phagocytosis, oxidative burst and intracellular killing of Candida albicans were analyzed by flow cytometry . Neither caspofungin nor anidulafungin significantly influenced phagocytosis . Only caspofungin significantly influenced oxidative burst after 15 min of incubation ( P<0.05) . Both caspofungin and anidulafungin improved intracellular killing rates of C . albicans after 2 h of incubation (42.4% and 43.2%, respectively, compared to 37.9% in controls; P<0.05) . In conclusion, caspofungin significantly improves oxidative burst and intracellular killing, which may be advantageous for clinical therapy . Pharmazie, 2004 Jul, 59(7), 573 - 4 The efficiency of the benzothiazole APB, the echinocandin micafungin, and amphotericin B in fluconazole-resistant Candida albicans and Candida dubliniensis; Melkusova S et al.; This study presents the efficiency of the experimental antifungal agents 6-amino-2-n-pentylthiobenzothiazole (APB) and the echinocandin micafungin, and amphotericin B against fluconazole-resistant Candida albicans and Candida dubliniensis (MIC95 for fluconazole > 64 mg/l) . The benzothiazole APB was less active against C . albicans and C . dubliniensis (MIC80 = 8 - 32 mg/l, MIC95 = 16 - 64 mg/l) than amphotericin B, which was efficient in a concentration range from 0.125 to 2 mg/l . However, the efficiency of micafungin was very high with MIC80, and MIC100 < or = 0.031 mg/l. Mikrobiyol Bul, 2004 Jan-Apr, 38(1-2), 99 - 103 {Esterase activities of Candida species}; Keceli SA et al.; The aim of this study was to determine the esterase activity of clinically important Candida species and to evaluate tween 80 opacity test medium as an additional identification method . A total of 118 Candida isolates (86 C . albicans, 12 C . tropicalis, 5 C . glabrata, 4 C . krusei, 3 C . guilliermondii, 3 C . kefyr, 3 C . parapsilosis, 1 C . famata, 1 C . dubliniensis) were examined for their response to tween 80 opacity test . Our results showed that, 83 of 86 (96.5%) Candida albicans, and all of the C . tropicalis and C . guilliermondii strains (100%) yielded a halo around their colonies . The remaining Candida species did not produce a positive test response after 13 days of incubation . It can be concluded that, tween 80 opacity test is a simple and economical test that can be used as a supplementary test for the identification of Candida species. Mund Kiefer Gesichtschir, 2004 Jul, 8(4), 261 - 3 Epub 2004 May 14. {Shammah-associated oral leukoplakia-like lesions}; Bethke G et al.; CASE REPORT: Shammah is a chewing tobacco, commonly used in Northern Africa . Leukoplakia-like lesions and oral cancer may be induced . In a 73-year-old male patient from Algeria leukoplakia-like lesions were observed in the anterior mandibular vestibulum and lower lip . The patient has been using shammah for 39 years . During the day three portions of shammah wrapped in a piece of paper tissue are prepared and rest in situ for 4-5 h . Due to its high alkalinity, shammah induces lesions resembling a burn . FINDINGS: Clinically, a white homogeneous lesion was seen in the vestibulum and mucosa of the lower lip . The white lesions could not be wiped off . Gingival recessions were seen in the lower front teeth . Root surfaces showed black-brown discoloration . A brush biopsy did not reveal epithelial atypia . The oral cavity showed signs of denture stomatitis and erythematous candidiasis . Microbiologically, Candida albicans was demonstrated . CONCLUSION: Due to the possibility of oral cancer in association with this habit, leukoplakia-like lesions have to be followed up diligently . Since chewing tobacco is a rare habit in Germany, changes as observed in the present case report may only rarely be observed . Due to migration it seems likely that in the future oral habits may be observed which are atypical for Western Europe . This phenomenon has also been addressed as transcultural dentistry. J Infect Chemother, 2004 Jun, 10(3), 181 - 4 Augmented inhibition of Candida albicans growth by murine neutrophils in the presence of a tryptophan metabolite, picolinic acid; Abe S et al.; The effects of picolinic acid (PLA), a product of tryptophan catabolism, on anti- Candida activity of neutrophils were studied . Casein-induced peritoneal neutrophils of C3H/He mice partially inhibited mycelial growth of Candida albicans when cultured with C . albicans for 16 h in vitro . The growth inhibition of Candida was augmented by a combination of neutrophils and more than 4 mM picolinic acid . Especially in the presence of 200 U/ml murine interferon-gamma (IFN-gamma), 2 mM picolinic acid augmented the anti- Candida activity of neutrophils . The physiological significance of the augmenting effects of picolinic acid is discussed. Microbiology, 2004 Aug, 150(Pt 8), 2641 - 51 The Candida albicans pH-regulated KER1 gene encodes a lysine/glutamic-acid-rich plasma-membrane protein that is involved in cell aggregation; Galan A et al.; Immunoscreening of a Candida albicans cDNA library with a polyclonal germ-tube-specific antibody (pAb anti-gt) resulted in the isolation of a gene encoding a lysine/glutamic-acid-rich protein, which was consequently designated KER1 . The nucleotide and deduced amino acid sequences of this gene displayed no significant homology with any other known sequence . KER1 encodes a 134 kDa lysine (14.5%)/glutamic acid (16.7%) protein (Ker1p) that contains two potential transmembrane segments . KER1 was expressed in a pH-conditional manner, with maximal expression at alkaline pH and lower expression at pH 4.0, and was regulated by RIM101 . A Deltaker1/Deltaker1 null mutant grew normally but was hyperflocculant under germ-tube-inducing conditions, yet this behaviour was also observed in stationary-phase cells grown under other incubation conditions . Western blotting analysis of different subcellular fractions, using as a probe a monospecific polyclonal antibody raised against a highly antigenic domain of Ker1p (pAb anti-Ker1p), revealed the presence of a 134 kDa band in the purified plasma-membrane fraction from the wild-type strain that was absent in the homologous preparation from Deltaker1/Deltaker1 mutant . The pattern of cell-wall protein and mannoprotein species released by digestion with beta-glucanases, reactive towards pAbs anti-gt and anti-Ker1p, as well as against concanavalin A, was also different in the Deltaker1/Deltaker1 mutant . Mutant strains also displayed an increased cell-surface hydrophobicity and sensitivity to Congo red and Calcofluor white . Overall, these findings indicate that the mutant strain was affected in cell-wall composition and/or structure . The fact that the ker1 mutant had attenuated virulence in systemic mouse infections suggests that this surface protein is also important in host-fungus interactions. Curr Opin Microbiol, 2004 Aug, 7(4), 336 - 41 From commensal to pathogen: stage- and tissue-specific gene expression of Candida albicans; Hube B; Candida albicans is both a successful commensal and pathogen of humans that can infect a broad range of body sites . The transition from commensalism to parasitism requires a susceptible host but it is also an active process . Gene expression of C . albicans is regulated by an interplay between host and pathogen and at least one transcriptional program associated with the yeast-to-hyphal transition . This not only allows immediate adaptation to changing environmental conditions, but also prepares cells for subsequent steps of infection . Recent work using transcript profiling has begun to shed light on infection strategies of pathogenic fungi. Scand J Infect Dis, 2004, 36(5), 372 - 9 A multicenter, open-label clinical study of micafungin (FK463) in the treatment of deep-seated mycosis in Japan; Kohno S et al.; The efficacy and safety of micafungin (FK463), which is a new lipopeptide antifungal agent of the echinocandin class and is active against both Aspergillus and Candida species, were investigated in patients with deep-seated mycosis in this study . 70 patients were treated with micafungin 12.5-150 mg/d intravenously for up to 56 d . The overall clinical response rates were 60% (6/10) in invasive pulmonary aspergillosis, 67% (6/9) in chronic necrotizing pulmonary aspergillosis, 55% (12/22) in pulmonary aspergilloma, 100% (6/6) in candidemia, and 71% (5/7) in esophageal candidiasis . The response rates for patients with prior antifungal treatment which was considered ineffective or toxic, were similar to rates for patients without prior treatment . Mycological eradication was observed in patients infected with Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Aspergillus niger, Candida albicans, Candida glabrata, or Candida krusei . Adverse events related to micafungin were reported in 21 patients (30%), and there was no dose-related occurrence of any adverse event . It is concluded that treatment with micafungin as monotherapy seems to be effective and safe in patients with deep-seated mycosis. Med Mycol, 2004 Jun, 42(3), 267 - 71 Recurrent isolation of an uncommon yeast, Candida pararugosa, from a sarcoma patient; Nakagawa Y et al.; A yeast was repeatedly isolated from the saliva of a sarcoma patient . A relatively uncommon species, Candida maris, was identified based on the API 20C profile . The yeast species most frequently obtained from the patient's mother and from clinic staff was Candida albicans . A comparison of the yeast obtained from the patient with the type strain of C . maris strongly suggested that the former was not representative of C . maris . Analysis of partial ribosomal DNA sequences of the patient strain and from the type strain of C . maris showed that the two are phylogenetically not closely related . The patient strain was very close to Candida pararugosa, a relatively uncommon asporogenous yeast . DNA reassociation studies among C . pararugosa and patient isolates showed that they were conspecific . We could not determine the source of the yeast infection . This case will alert hospital staff to be aware of the possibility of unexpected environmental microorganisms as causes of infections, colonizations and persistent environmental contamination events in immunocompromised patients. Nippon Ishinkin Gakkai Zasshi, 2004, 45(3), 131 - 6 {Cytokine production and dermatophytosis}; Kano R; The characteristic pathological feature of dermatomycosis is numerous neutrophilic infiltrates within the epidermis . However, the precise mechanism of this infiltration remains unknown . In this study, interleukins 1 beta, 6, and 8, monocyte chemotactic protein-1 (MCP-1), and tumor necrosis factor (TNF)-alpha levels in the medium where keratinocytes were co-cultured with Candida albicans, Malassezia and Trichophyton mentagrophytes, were determined by enzyme-linked immunosorbent assays (ELISAs) in order to estimate the effect of these fungi on the cytokine production from human keratinocytes . The IL-8 level in the supernatants increased with 1 to 14 hours of co-culture in response to live C . albicans, but the other cytokines were undetectable . Furthermore, the mRNA of IL- 8 in keratinocytes was also confirmed to increased . This data suggested that C . albicans directly induce interleukin 8 production from human keratinocytes without activated macrophages . The IL-6, IL-8, and TNF-alpha levels in the culture supernatants increased with 1 to 24 hours of co-culture with keratinocytes and Malassezia species but the MCP-1 level was undetectable . The IL-8 and TNF-alpha levels in the culture supernatants increased with 1 to 24 hours of co-culture with keratinocytes and Trichophyton mentagrophytes but the other cytokine levels were undetectable . The ELISA analysis of cytokine production by human keratinocytes will provide useful information in understanding the pathogenesis of dermatomycosis. Med Mycol, 2004 Jun, 42(3), 283 - 5 Antifungal susceptibility of 50 Candida isolates from invasive mycoses in Chile; Silva V et al.; We determined the antifungal susceptibility of 50 Candida isolates from invasive mycoses in intensive care unit patients in Chile . Candida albicans (27 isolates) and C . parapsilosis (12 isolates) were the most common etiologic species . Candida glabrata (five isolates) was isolated only from children . Our data are consistent with those of recent Brazilian and Argentinean studies but differ from those of some US, Canadian and Norwegian studies, in which the relatively azole-resistant C . glabrata was the predominant non-C . albicans species seen . All isolates in this study were susceptible to amphotericin B . Itraconazole and fluconazole resistance were observed in 6 and 4% of the isolates, respectively. Med Mycol, 2004 Jun, 42(3), 255 - 60 Reservoir of Candida albicans infection in a vascular bypass graft demonstrates a stable karyotype over six months; Lephart P et al.; We retrospectively analyzed five Candida albicans isolates from two infection episodes in a single patient 6 months apart . Using contour-clamped homogeneous field electrophoresis (CHEF), random amplified polymorphic DNA (RAPD) fingerprinting, and restriction fragment length polymorphism (RFLP) complex probe 27A as means of molecular typing, we demonstrate an unvarying genotype amongst the infection-causing C . albicans strains . Several months later, the patient yielded C . glabrata in a yeast survey of oral and rectal sites . The preponderance of C . glabrata and lack of C . albicans isolated from normal flora sites suggests that this patient harbored the prior C . albicans bloodstream isolate on a Gore-Tex graft for 6 months prior to the second episode of fungemia. Fogorv Sz, 2004 Jun, 97(3), 123 - 7 {Microbiological testing of the artificial gingival margin in dentures}; Hermann P et al.; In everyday practice dental laboratories try to reproduce the natural form of sulcus gingivae at the transitional area between artificial teeth and gingiva of removable dentures, even on esthetically less important areas . Aim of these investigations were to examine how artificial recreation of the sulcus gingivae influences plaque retention, and what is the microbiological relevance of these . Investigations were carried out on the vestibular side of removable dentures of 32 randomly selected patients treated at the Department of Prosthodontics at the Faculty of Dentistry, Semmelweis University . Microbiological samples were taken from each patient using the same method . Samples were taken from the left upper first molars' artificial gingival margin using sterile paper points . Paper points were then transported in Eppendorf-tubes, in 2 ml of physiological saline solution, and processed within a two-hour period of time . Series dilutions were made of the sample solutions, then surface-streaked on Subaraud and Gentamycin, blood-agar, eosin-methylene blue and Mitis Salivarius culture enriched with Bacitracin . Subaraud culture was induced under aerob conditions, at room temperature for two days, then the total amount of fungi quantified . After pure-culturing Candida albicans ID-culture was used for identification, and BioMerieux ATB automatic equipment to identify different Candida species . From pure cultures identification was carried out with Gram-staining, Neisser-staining, catalase, oxidase and also with other biochemical reactions . Blood-agar was used to determine total germ count, and normal commensal pharyngeal and oral bacteria . After collecting the microbiological samples, the conventional shape of the dental margin of gingiva was abolished on one side of the dentures and a smooth transition was created between denture teeth and the artificial gingiva in the molar and premolar region . During our investigations only blastomycetes were found . Besides most common Candida albicans, Candida glabrata, tropicallis, spherica and lambica were also identified . Our patients did not miss artificial sulcus gingivae, had no aesthetic complaints about smooth transition between artificial teeth and gingiva . Microbiological investigations of the samples and the comparative analysis showed, that on the smooth transitional areas compared to conventionally shaped sulci significantly less gram-negative, gram-positive bacteria and oral fungi were found, and there was no plaque formation. J Antimicrob Chemother, 2004 Sep, 54(3), 603 - 8 Epub 2004 Jul 28. Release of calcium from intracellular stores and subsequent uptake by mitochondria are essential for the candidacidal activity of an N-terminal peptide of human lactoferrin; Lupetti A et al.; OBJECTIVES: Earlier studies showed that mitochondrial damage is a hallmark of the candidacidal activity of an N-terminal peptide of human lactoferrin, further referred to as hLF(1-11) . Since uptake of Ca(2+) by mitochondria may be essential for their activation, the aim of this study was to define the role of Ca(2+) in killing of Candida albicans by the hLF(1-11) peptide . METHODS: The effect of compounds interfering with Ca(2+) homeostasis on the hLF(1-11)-induced candidacidal activity, changes in mitochondrial membrane potential, and reactive oxygen species production were evaluated using a killing assay, rhodamine 123 staining, and 2',7'-dichlorofluorescein diacetate, respectively . The increase in cellular Ca(2+) content was measured using (45)Ca(2+) . RESULTS: Our results revealed that Ruthenium Red, which inhibits the mitochondrial Ca(2+)-uniporter and the voltage-sensitive Ca(2+) release from internal stores, blocked (P<0.05) the hLF(1-11)-induced candidacidal activity as well as changes in the membrane potential of mitochondria, and reactive oxygen species production . Oxalate, which precipitates Ca(2+) in intracellular organelles, decreased (P<0.05) the peptide-induced changes in the membrane potential of mitochondria, reactive oxygen species production, and candidacidal activity . Furthermore, the Ca(2+) ionophore ionomycin combined with high CaCl(2) concentrations enhanced the hLF(1-11)-induced candidacidal activity . Moreover, hLF(1-11) caused an influx of Ca(2+) from the extracellular medium into C . albicans reaching a three-fold increase at 2 h, whereas no increase was found in unexposed cells . In agreement, the Ca(2+)-chelator EGTA blocked the peptide-induced candidacidal activity . CONCLUSIONS: Ca(2+) release from intracellular stores, probably through subsequent mitochondrial Ca(2+) uptake, is essential for the hLF(1-11)-induced candidacidal activity. Clin Experiment Ophthalmol, 2004 Aug, 32(4), 441 - 3 Churg-Strauss syndrome presenting with conjunctival nodules in association with Candida albicans and ankylosing spondylitis; Ooi KG et al.; Churg-Strauss syndrome is a rare diffuse vasculitis of which the ocular manifestations have been well documented . However, reports of conjunctival involvement in Churg-Strauss syndrome are scarce . Such a presentation is described in a man with Candida albicans infection as well as ankylosing spondylitis, and a possible aetiological linkage is established amongst all three. Med Sci Monit, 2004 Aug, 10(8), BR239 - 49 Epub 2004 Jul 23. Involvement of interleukin-18 in the inflammatory response against oropharyngeal candidiasis; Tardif F et al.; BACKGROUND: Oral candidiasis is a collective name for a group of disorders caused by the dimorphic fungus Candida albicans (C . albicans) . Host defenses against C . albicans essentially fall into two categories: specific immune mechanisms and local oral mucosal epithelial cell defenses . The rationale of this study was to investigate the involvement of IL-18 in the inflammatory response against oral candidiasis . MATERIAL/METHODS: We first used human oral mucosa tissue and saliva to assess the production of Il-18 . Second, we engineered human oral mucosa using only normal human oral epithelial cells and fibroblasts . Tissues were infected with C . albicans at different time points . RESULTS: Tissue and saliva analyses demonstrated that constitutively produced and secreted IL-18 was up-regulated following Candida-infection . With our engineered model, we showed that C . albicans significantly increased the secretion of active IL-18 by infected epithelial cells . Interestingly, a significant secretion of IFNg functionally supported the up-regulation of active IL-18 in C . albicans-infected tissues . We also showed that rhIL-18 increased the expression and production of endogenous IL-18 and ICE in C . albicans-infected tissues, which was paralleled by a significant increase in IFNg secretion . CONCLUSIONS: These data suggest that (i) oral epithelial cells are involved in local host defenses against C . albicans infections, via IFNg induced-IL-18, and (ii) that IL-18 and IFNg secretions may be related to epithelial cells . Given that our experimental model closely mimics the natural interface between the oral mucosa and C . albicans, it appears that IL-18 meets the requirements of being a cytokine that epithelial cells use to control C . albicans infections. J Biol Chem, 2004 Sep 24, 279(39), 40861 - 7 Epub 2004 Jul 23. SUMO modification of septin-interacting proteins in Candida albicans; Martin SW et al.; The initiation of bud and hyphal growth in the opportunistic fungal pathogen Candida albicans both involve polarized morphogenesis . However, there are many differences including the function of the septin proteins, a family of proteins involved in membrane organization in a wide range of organisms . Septins form a characteristic ring on the inner surface of the plasma membrane at the bud neck, whereas the septins are diffusely localized across emerging hyphal tips . In addition, septin rings are maintained at sites of septum formation in hyphae rather than being disassembled immediately after cytokinesis . The possibility that C . albicans septins are regulated by the small ubiquitin-like protein SUMO was examined in this study because the Saccharomyces cerevisiae septins were shown previously to be modified by SUMO (Smt3p) . However, SUMO conjugation to septins was not detected during budding or hyphal morphogenesis in C . albicans . These results are supported by the lack of conserved SUMO consensus motifs between septins from the two organisms even after adjusting the predicted Cdc3p and Cdc12p septin sequences to account for mRNA splicing in C . albicans . Interestingly, a homolog of the Smt3p SUMO was identified in the C . albicans genome, and an epitope tagged version of Smt3p was conjugated to a variety of proteins . Immunofluorescence analysis showed prominent Smt3p SUMO localization at bud necks and sites of septum formation in hyphae similar to the septins . However, Smt3p was primarily detected on the mother cell side of the septin ring . A subset of these Smt3p-modified proteins co-immunoprecipitated with the septin Cdc11p . These results indicate that septin-associated proteins and not the septins themselves are the key target of SUMO modification at the bud neck in C . albicans . Indian J Exp Biol, 2004 Jan, 42(1), 86 - 90 Effect of nitric oxide on H+ -efflux in presence of various nutrients in Candida albicans; Haque MM et al.; In the present study tentative link has been established between H+ -efflux and effect of NO in presence of various nutrients (glucose, 2-deoxy-D-glucose, xylose, proline, glutamic acid and lysine) in C . albicans using sodium nitroprusside (SNP) as a potent source of NO . It was observed that there was a decreasing trend in pH with time, in control, while SNP treated cells showed an initial decline in pH for 10-15 min, followed by an increase in pH up to 30 min . In presence of glucose there was an enhancement in H+ -efflux by 9-fold whereas proline, glutamic acid and lysine showed enhancement by 3, 6 and 1.5-fold respectively . Similar trends in increase in pH after 15 min in SNP treated cells of Candida was observed in presence of all nutrients used . It was demonstrated for the first time that H+ -ATPase of C . albicans was affected by NO. Proteomics . 2004 Aug;4(8):2513. Contribution of the antibodies response induced by a low virulent Candida albicans strain in protection against systemic candidiasis (vol . 4, Issue 4, pp . 1204-1215); Fernandez-Arenas E et al.; No abstracts.http://dx.doi.org/10.1002/pmic.200300678 Proteomics, 2004 Aug, 4(8), 2425 - 36 Proteomic response to amino acid starvation in Candida albicans and Saccharomyces cerevisiae; Yin Z et al.; Saccharomyces cerevisiae activates general amino acid control (GCN) in response to amino acid starvation . Some aspects of this response are known to be conserved in other fungi including Candida albicans, the major systemic fungal pathogen of humans . Here, we describe a proteomic comparison of the GCN responses in S . cerevisiae and C . albicans . We have used high-resolution two-dimensional (2-D) gel electrophoresis and peptide mass fingerprinting to develop a 2-D protein map of C . albicans . A total of 391 protein spots, representing 316 open reading frames, were identified . Fifty-five C . albicans and 65 S . cerevisiae proteins were identified that responded reproducibly to 3-aminotriazole (3AT) in a Gcn4p-dependent fashion . The changes in the S . cerevisiae proteome correlated with the response in the S . cerevisiae transcript profile to 3AT treatment (rank correlation coefficient = 0.59; Natarajan et al., Molec . Cell . Biol . 2001, 21, 4347-4368) . Significant aspects of the GCN response were conserved in C . albicans and S . cerevisiae . In both fungi, amino acid biosynthetic enzymes on multiple metabolic pathways were induced by 3AT in a Gcn4p-dependent fashion . Carbon metabolism functions were also induced . However, subtle differences were observed between these fungi . For example, purine biosynthetic enzymes were induced in S . cerevisiae, but were not significantly induced in C . albicans . These differences presumably reflect the contrasting niches of these relatively benign and pathogenic yeasts, respectively. Antimicrob Agents Chemother, 2004 Aug, 48(8), 3064 - 79 Comparison of gene expression profiles of Candida albicans azole-resistant clinical isolates and laboratory strains exposed to drugs inducing multidrug transporters; Karababa M et al.; Azole resistance in Candida albicans can be due to upregulation of multidrug transporters belonging to ABC (ATP-binding cassette) transporters (CDR1 and CDR2) or major facilitators (CaMDR1) . Upregulation of these genes can also be achieved by exposure to fluphenazine, resulting in specific upregulation of CDR1 and CDR2 and by exposure to benomyl, resulting in specific CaMDR1 upregulation . In this study, these two different states of gene upregulation were used to determine coregulated genes that often share similar functions or similar regulatory regions . The transcript profiles of a laboratory strain exposed to these drugs were therefore determined and compared with those of two matched pairs of azole-susceptible and -resistant strains expressing CDR1 and CDR2 (CDR strains) or CaMDR1 (MDR isolates) . The results obtained revealed that, among 42 commonly regulated genes (8.6% of all regulated genes) between fluphenazine-exposed cells and CDR isolates, the most upregulated were CDR1 and CDR2 as expected, but also IFU5, RTA3 (which encodes putative membrane proteins), HSP12 (which encodes heat shock protein), and IPF4065 (which is potentially involved in stress response) . Interestingly, all but HSP12 and IPF4065 contain a putative cis-acting drug responsive element in their promoters . Among the 57 genes (11.5% of all regulated genes) commonly regulated between benomyl-exposed cells and MDR isolates, the most upregulated were CaMDR1 as expected but also genes with oxido-reductive functions such as IFD genes, IPF5987, GRP2 (all belonging to the aldo-keto reductase family), IPF7817 {NAD(P)H oxido-reductase}, and IPF17186 . Taken together, these results show that in vitro drug-induced gene expression only partially mimics expression profiles observed in azole-resistant clinical strains . Upregulated genes in both drug-exposed conditions and clinical strains are drug resistance genes but also genes that could be activated under cell damage conditions. Antimicrob Agents Chemother, 2004 Aug, 48(8), 3051 - 6 Susceptibility to thrombin-induced platelet microbicidal protein is associated with increased fluconazole efficacy against experimental endocarditis due to Candida albicans; Yeaman MR et al.; Platelet microbicidal proteins (PMPs) are believed to be integral to host defense against endovascular infection . We previously demonstrated that susceptibility to thrombin-induced PMP 1 (tPMP-1) in vitro negatively influences Candida albicans virulence in the rabbit model of infective endocarditis (IE) . This study evaluated the relationship between in vitro tPMP-1 susceptibility (tPMP-1s) or resistance (tPMP-1r) and efficacy of fluconazole (FLU) therapy of IE due to C . albicans . Candida IE was established in rabbits with either tPMP-1s or tPMP-1r strains . Treatment groups received FLU (100 mg/kg/day) intraperitoneally for 7 or 14 days; control animals received no therapy . At these time points, cardiac vegetations, kidneys, and spleens were quantitatively cultured to assess fungal burden . At both 7 and 14 days and in all target tissues, the extent of candidal clearance by FLU was greater in animals infected with the tPMP-1s strain than in those infected with the tPMP-1r strain . These differences were statistically significant in the spleen and kidney . Corroborating these in vivo data, FLU (a candidastatic agent), in combination with tPMP-1, exerted an enhanced fungicidal effect in vitro against tPMP-1s and tPMP-1r C . albicans, with the extent of this effect greatest against the tPMP-1s strain . Collectively, these results support the concept that tPMP-1 susceptibility contributes to the net efficacy of FLU against C . albicans IE in vivo, particularly in tissues in which platelets and tPMP-1 likely play significant roles in host defense. Proc Natl Acad Sci U S A, 2004 Aug 3, 101(31), 11374 - 9 Epub 2004 Jul 22. Centromeric DNA sequences in the pathogenic yeast Candida albicans are all different and unique; Sanyal K et al.; In an approach to clone and characterize centromeric DNA sequences of Candida albicans by chromatin immunoprecipitation, we have used antibodies directed against an evolutionarily conserved histone H3-like protein, CaCse4p (CENP-A homolog) . Sequence analysis of clones obtained by this procedure reveals that only eight relatively small regions (approximately 3 kb each) of the Can . albicans genome are selectively enriched . These CaCse4-bound sequences are located within 4- to 18-kb regions lacking ORFs and occur once in each of the eight chromosomes of Can . albicans . Binding of another evolutionarily conserved kinetochore protein, CaMif2p (CENP-C homolog), colocalizes with CaCse4p . Deletion of the CaCse4p-binding region of chromosome 7 results in a high rate of loss of the altered chromosome, confirming that CaCse4p, a centromeric histone in the CENP-A family, indeed identifies the functional centromeric DNA of Can . albicans . The CaCse4p-rich regions not only lack conserved DNA motifs of point (<400 bp) centromeres and repeated elements of regional (>40 kb) centromeres, but also each chromosome of Can . albicans contains a different and unique CaCse4p-rich centromeric DNA sequence, a centromeric property previously unobserved in other organisms. J Biol Chem, 2004 Sep 17, 279(38), 39628 - 35 Epub 2004 Jul 22. Loss of cell wall mannosylphosphate in Candida albicans does not influence macrophage recognition; Hobson RP et al.; The outer layer of the cell wall of the human pathogenic fungus Candida albicans is enriched with heavily mannosylated glycoproteins that are the immediate point of contact between the fungus and cells of the host, including phagocytes . Previous work had identified components of the acid-labile fraction of N-linked mannan, comprising beta-1,2-linked mannose residues attached via a phosphodiester bond, as potential ligands for macrophage receptors and modulators of macrophage function . We therefore isolated and disrupted the CaMNN4 gene, which is required for mannosyl phosphate transfer and hence the attachment of beta-1,2 mannose oligosaccharides to the acid-labile N-mannan side chains . With the mannosylphosphate eliminated, the mnn4Delta null mutant was unable to bind the charged cationic dye Alcian Blue and was devoid of acid-labile beta-1,2-linked oligomannosaccharides . The mnn4Delta mutant was unaffected in cell growth and morphogenesis in vitro and in virulence in a murine model of systemic C . albicans infection . The null mutant was also not affected in its interaction with macrophages . Mannosylphosphate is therefore not required for macrophage interactions or for virulence of C . albicans. Infect Immun, 2004 Aug, 72(8), 4878 - 80 Genetic susceptibility of mice to Candida albicans vaginitis correlates with host estrogen sensitivity; Clemons KV et al.; We compared susceptibility to Candida vaginitis in derived murine substrains differing in sensitivity to estrogen (CD-1 and CD10, resistant; CD3 and C57BL/6 responsive), and in F1 crosses . The order of decreasing resistance was CD-1 > or = CD10 > or = CD10 x CD3F1 > CD10 x B6F1 > CD3 > C57BL/6 and correlated with estrogen responsiveness in endocrine disruptor assays . Resistance to Candida vaginitis appears additive in CD10 x B6F1 animals and dominant in CD10 x CD3F1 animals. J Inorg Biochem, 2004 Aug, 98(8), 1361 - 6 Synthesis, X-ray crystal structure, anti-fungal and anti-cancer activity of {Ag2(NH3)2(salH)2} (salH2=salicylic acid); Coyle B et al.; {Ag(2)(NH(3))(2)(salH)(2)} (salH(2)=salicylic acid) was synthesised from salicylic acid and Ag(2)O in concentrated aqueous NH(3) and the dimeric Ag(I) complex was characterised using X-ray crystallography . The complex is centrosymmetric with each metal coordinated to a salicylate carboxylate oxygen and to an ammonia nitrogen atom in an almost linear fashion . The two {Ag(NH(3))(salH)} units in the complex are linked by an Ag-Ag bond . Whilst metal-free salH(2) did not prevent the growth of the fungal pathogen Candida albicans {Ag(2)(NH(3))(2)(salH)(2)}, {Ag(2)(salH)(2)} and some simple Ag(I) salts greatly inhibited cell reproduction . SalH(2), {Ag(2)(NH(3))(2)(salH)(2)} {Ag(2)(salH)(2)} and AgClO(4) produced a dose-dependent cytotoxic response against the three human derived cancer cell lines, Cal-27, Hep-G2 and A-498, with the Ag(I)-containing reagents being the most effective. J Nat Prod, 2004 Jul, 67(7), 1131 - 4 Quinazolin-4-one derivatives from Streptomyces isolates; Maskey RP et al.; From the ethyl acetate extract of the strain Streptomyces sp . isolate GW23/1540, besides 16 known products, several 1H-quinazolin-4-one derivatives were isolated . (SR)-2-(1-Hydroxyethyl)-2-methyl-2,3-dihydro-1H-quinazolin-4-one (4) and (RR)-2-(1-hydroxyethyl)-2-methyl-2,3-dihydro-1H-quinazolin-4-one (5) are new natural products . 2-Methyl-3H-quinazolin-4-one (2) and 1H-quinazoline-2,4-dione (3) are known from other bacteria and plants, respectively . From another Streptomyces sp., GW2/577, 5-methyl-1H-quinazoline-2,4-dione (6) was isolated and the structure proven by comparison with the isomeric 7 . The new natural products showed no activity against the microalgae Chlorella vulgaris, Chlorella sorokiniana, and Scenedesmus subspicatus, the fungus Mucor miehei, the yeast Candida albicans, and the bacteria Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Streptomyces viridochromogenes. Mol Biol Cell, 2004 Oct, 15(10), 4490 - 9 Epub 2004 Jul 21. Transcription profiling of cyclic AMP signaling in Candida albicans; Harcus D et al.; We used transcription profiling in Candida albicans to investigate cellular regulation involving cAMP . We found that many genes require the adenylyl cyclase Cdc35p for proper expression . These include genes encoding ribosomal subunit proteins and RNA polymerase subunit proteins, suggesting that growth could be controlled in part by cAMP-mediated modulation of gene expression . Other genes influenced by loss of adenylyl cyclase are involved in metabolism, the cell wall, and stress response and include a group of genes of unknown function that are unique to C . albicans . The profiles generated by loss of the adenylyl cyclase regulator Ras1p and a downstream effector Efg1p were also examined . The loss of Ras1p function disturbs the expression of a subset of the genes regulated by adenylyl cyclase, suggesting both that the primary role of Ras1p in transcriptional regulation involves its influence on the function of Cdc35p and that there are Ras1p independent roles for Cdc35p . The transcription factor Efg1p is also needed for the expression of many genes; however, these genes are distinct from those modulated by Cdc35p with the exception of a class of hyphal-specific genes . Therefore transcription profiling establishes that cAMP plays a key role in the overall regulation of gene expression in C . albicans, and enhances our detailed understanding of the circuitry controlling this regulation. Zhonghua Gan Zang Bing Za Zhi, 2004 Jul, 12(7), 389 - 91 {Clinical study on the severe hepatitis with nosocomial fungal infections and risk factors}; Zhang XH et al.; OBJECTIVE: To study the nosocomial fungal infections in the patient with severe hepatitis and analyze of risk factor . METHODS: All 115 severe hepatitis with fungal infections inpatients was studied prospectively . RESULTS: We identified 115 cases with fungal infections, the mean age of patients was 37.2+/-21.5 years, male: 49 cases, female 66 cases . Infection of abdominal cavity accounted for 40.9%, infectious rate in respiratory tract and digestive tract were 26.9%, 21.8%, respectively . Candida albicans accounted for 67.6% . Use of broad-spectrum antibiotic and corticosteroids, neutropenia, severity of liver disease, improper medical manipulations as significant risk factors for fungal infection . Death rate of study group and control group was 59.1%, 34.8%, respectively (x2=36.0) . In multivariate analysis, neutropenia, disseminated infection and severity of liver diseases were independent prognostic factors . CONCLUSION: Identification of risk factors and predictors of a poor outcome in patients with severe hepatitis with fungal infections, it suggested that implications in prophylaxis of fungal infection, early diagnosis and appropriate therapy would be important for these patients. Eur Rev Med Pharmacol Sci, 2004 Mar-Apr, 8(2), 69 - 77 Fluconazole resistance in Candida albicans: a review of mechanisms; Casalinuovo IA et al.; Antifungal agents have greatly contributed to the improvement of public health . Nevertheless, antifungal resistant pathogens have increased during the past decade, becoming a serious concern . Candida albicans has been the most extensively studied pathogen in antifungal resistance because of their morbidity and mortality associated with infections in immunocompromised patients . This review describes the antifungal mechanims of the azole fluconazole widely used for the prophylaxis and treatment of candidal infections . The specific molecular pathways occurring in fluconazole-resistance of C . albicans and some issues about new antifungal agents are also discussed. Clin Exp Obstet Gynecol, 2004, 31(2), 113 - 6 The importance of diagnostic work-up in the management of candidal vulvovaginitis . A prospective study; Ben-Haroush A et al.; OBJECTIVE: To assess the accuracy of the diagnostic work-up in identifying vaginal candidal infection, and to determine the safety, efficacy and speed of action of clotrimazole vaginal tablets . PARTICIPANTS: Two hundred and twenty-three patients with symptoms and signs of candidal infection, a presence of vaginal pH < or = 4.5 and positive 10% KOH examination . METHODS: After vaginal culture was taken, the eligible patients were treated with clotrimazole 200 mg vaginal tabs (manufactured by Teva or Agis) . RESULTS: Cultures grew Candida albicans in 189 cases (85%), and Candida non-albicans in five (2.2%); 29 patients (13%) did not have any candidal infection . Of the 189 C . albicans-positive patients, 174 were reassessed for effects of clotrimazole treatment by self-reports and objective measures . Ninety-four percent of the patients reported improvement after treatment, rated moderate to high by 87% . The physician evaluation was similar: some improvement in 96%, and moderate or high improvement in 91% . At the second examination, 7.5% of the treated patients still had a positive culture for Candida albicans, and they remained positive on KOH microscopic examination, although vaginal pH was significantly higher . Maximal improvement was recorded three to four days after starting treatment . CONCLUSION: It is important that the diagnostic work-up for suspected candidal infections consist of at least vaginal pH measurement and microscopy study with KOH . We encourage the use of vaginal cultures, especially in recurrent cases . Clotrimazole is a safe and effective treatment. Biotechnol Lett, 2004 Jul, 26(14), 1125 - 30 Anti-fungal effects of phenolic amides isolated from the root bark of Lycium chinense; Lee DG et al.; Four phenolic amides, dihydro-N-caffeoyltyramine (1), trans-N-feruloyloctopamine (2), trans-N -caffeoyltyramine (3), and cis-N-caffeoyltyramine (4), were isolated from an ethyl acetate extract of the root bark of Lycium chinense Miller . All had an anti-fungal effect; compounds 1-3 were potent at 5-10 microg ml(-1) and were without hemolytic activity against human erythrocyte cells . Compound 4 was active at 40 microg ml(-1) . All four compounds impeded the dimorphic transition of pathogen, Candida albicans. Eur J Pharm Sci, 2004 Aug, 22(5), 451 - 8 Comparative therapeutic efficacy of a novel lyophilized amphotericin B lecithin-based oil-water microemulsion and deoxycholate-amphotericin B in immunocompetent and neutropenic mice infected with Candida albicans; Brime B et al.; The in vivo efficacy of a new amphotericin B (AmB) oil-in-water lecithin-based microemulsion delivery system (M-AmB) compared to deoxycholate-AmB (D-AmB) was studied in an immunocompetent and neutropenic murine model of systemic candidiasis . D-AmB was administered at the maximum tolerated dose of 1 mg/kg whereas M-AmB was given at the doses of 1, 2 and 3 mg/kg; doses were well tolerated due to their reduced toxicity . Both formulations were administered 24, 48 and 72 h after infection in immunocompetent mice, and 2, 6 and 24 h after infection in neutropenic mice . Kaplan-Meier survival curves showed that the M-AmB treated group had a better survival time than infected mice without treatment used as a control group (P = 4.66 x 10(-6)), and the Mann-Whitney W statistical test indicated that it reduced the percentage of mortality and fungal load in the most representative organs . This new formulation is a designed competitor which has proved to present better results than D-AmB in an established infection not only in immunocompetent but in neutropenic mice as well. J Biol Chem, 2004 Sep 24, 279(39), 40737 - 47 Epub 2004 Jul 15. Expression of transglutaminase substrate activity on Candida albicans germ tubes through a coiled, disulfide-bonded N-terminal domain of Hwp1 requires C-terminal glycosylphosphatidylinositol modification; Staab JF et al.; By serving as a microbial substrate for epithelial cell transglutaminase, Hwp1 (Hyphal wall protein 1) of Candida albicans participates in cross-links with proteins on the mammalian mucosa . Biophysical properties of the transglutaminase substrate domain were explored using a recombinant protein representative of the N-terminal domain of Hwp1 and were similar to other transglutaminase substrates, the small proline-rich proteins of cornified envelopes found in stratified squamous epithelia . Recombinant Hwp1 lacks alpha and beta structures by circular dichroism and likely exists as a disulfide-cross-linked coiled-coil . The transglutaminase substrate property prompted a unique approach for investigating the features of surface Hwp1 on germ tubes . A lysine analog, 5-(biotinamido)pentylamine, was cross-linked to germ tubes catalyzed by transglutaminase 2 prior to cell fractionation, immunoprecipitation, and detection with streptavidin conjugates . The majority of the transglutaminase-modifiable Hwp1 was covalently attached to the beta-glucan of hyphae by the C terminus of Hwp1 via a glycosylphosphatidylinositol remnant anchor . A putative precursor of cell wall forms of Hwp1 was identified in the cell extract and in the culture medium . Hwp1 was modified by relatively short N-linked glycans, and the molecular size of the protein was reduced by hypomannosylation when expressed in O-glycosylation mutant strains . Hwp1 combines features of mammalian transglutaminase substrate proteins with characteristics of fungal cell wall proteins to form an unconventional adhesin at the hyphal wall of C . albicans . J Control Release, 2004 Aug 11, 98(2), 269 - 79 Chitosan delivery systems for the treatment of oral mucositis: in vitro and in vivo studies; Aksungur P et al.; Oral mucositis is a frequent and potentially severe complication of radiation or chemotherapy for cancer . Associated with atrophy and ulceration of the oral mucosa is an increased risk of infection, and the most common pathogenic agent is Candida . Chitosan is an excellent candidate for the treatment of oral mucositis . Its bioadhesive and antimicrobial properties offer the palliative effects of an occlusive dressing and the potential for delivering drugs, including anti-candidal agents . The aim of this study was to develop an occlusive bioadhesive system for prophylaxis and/or treatment of oral mucositis . Gel and film formulations were prepared using chitosans at different molecular weights and in different solvents . Nystatin, which is considered as a prophylactic agent for oral mucositis was incorporated into the formulations . The in vitro release of nystatin from the formulations was decreased with the increasing molecular weight of chitosan . The effect of the formulations was investigated in vivo in hamsters with chemotherapy-induced mucositis . Mucositis scores in groups treated with nystatin incorporated into gel and suspension formulations were significantly lower (p < 0.05) than those treated with the chitosan gel alone . Survival of animals in the treated groups was higher than that in the control group . The retention time and distribution of the gels in the oral cavity were investigated in healthy volunteers . A faster distribution of nystatin in the oral cavity was obtained using the suspension compared to the gels, but the nystatin saliva level decreased rapidly as well . A drug concentration above the minimum inhibitory concentration (MIC) value for Candida albicans (0.14 microg/ml) was maintained for longer periods of time at the application site (90 min) than at the contralateral site (45 min) in the oral cavity. Carbohydr Res, 2004 Aug 2, 339(11), 1889 - 97 Sugar bislactones by one-step oxidative dimerisation with pyridinium chlorochromate versus regioselective oxidation of vicinal diols; Rauter AP et al.; Synthesis of 10-membered bislactones by PCC oxidation of methyl 2,6-di-O-pivaloyl-alpha-d-glucopyranoside and methyl 4,6-O-benzylidene-alpha-d-glucopyranoside is described, with emphasis on their structure elucidation using the information gained by combination of NMR spectroscopic techniques with X-ray diffraction data . In alternative, the use of PCC and PCC adsorbed on silica gel or alumina for the regioselective oxidation of vicinal diols in sugars is also reported . Both bislactones showed antifungal activity against Candida albicans, and were slightly active against the bacteria Bacillus subtilis . The bislactone presenting pivaloyl protecting groups also promoted some growth inhibition of Staphylococcus aureus. Biometals, 2004 Aug, 17(4), 415 - 22 Metal complexes of 1,10-phenanthroline-5,6-dione alter the susceptibility of the yeast Candida albicans to amphotericin B and miconazole; Eshwika A et al.; Growth of the pathogenic yeast Candida albicans in sub-MIC (minimum inhibitory concentration) levels of Cu(ClO4)2 6H2O and {Cu(phendio)3}(ClO4)2 4H2O (phendio = 1,10-phenanthroline-5,6-dione) increased the concentration of miconazole and amphotericin B required to achieve the MIC90 whereas pre-growth in AgClO4 and {Ag(phendio)2}ClO4 resulted in a small decrease in the relevant MIC90 values . The copper complexes reduce the oxygen consumption of C . albicans while the silver complexes increase oxygen consumption . In addition, pre-growth of cells in the copper complexes resulted in a lower ergosterol content while the silver complexes induced an elevation in ergosterol synthesis . The ability of copper and silver complexes to alter the susceptibility of C . albicans to miconazole and amphotericin B may be influenced by their action on respiration, since reduced respiration rates correlate with reduced cellular ergosterol which is the target for amphotericin B . Lower levels of ergosterol have previously been associated with elevated tolerance to this drug . In the case of reduced sensitivity to miconazole, tolerance may be mediated by lower ergosterol synthesis giving rise to fewer toxic side products once biosynthesis is inhibited by miconazole. Minerva Ginecol, 2004 Apr, 56(2), 149 - 53 {Preliminary results of the Italian epidemiological study on vulvo-vaginitis}; Boselli F et al.; AIM: The aim of this work was to evaluate the prevalence of symptomatic vulvo-vaginal infections among the Italian female population and the diagnostic approach of 158 Italian gynaecologists . METHODS: A total of 1644 patients were enrolled in this survey . A presumed diagnosis of vulvovaginal infection was made according to specific clinical and laboratory criteria (pH e sniff test) in 902 (55.4%) cases, whereas a definitive diagnosis was made in 1439 (87.5%) cases . The definitive diagnoses were as follows: 844 (51.3%) vulvovaginal mycosis, 327 (19.9%) bacterial vaginosis, 110 (6.7%) trichomonal infection, 100 (6.1%) aspecific bacterial vaginitis, 58 (3.5%) non-infectious vaginitis . As assessed by typing, mycosis were mainly due to Candida albicans infections in 459 cases (78%) . CONCLUSION: From the survey it is possible to infer that: 1) only thanks to the employed microbiological diagnostic tests a definitive diagnosis was made in 702 patients without a previous presumed diagnosis; 2) mycotic infections were underestimated; 3) Candida albicans was the most common species causing female low genital tract mycotic infections . In conclusion, these data underline the importance of laboratory examinations in the diagnosis of low genital tract infections for the ambulatory gynaecological practice. Microbiology, 2004 Jul, 150(Pt 7), 2475 - 89 An aspartic proteinase gene family in the filamentous fungus Botrytis cinerea contains members with novel features; ten Have A et al.; Botrytis cinerea, an important fungal plant pathogen, secretes aspartic proteinase (AP) activity in axenic cultures . No cysteine, serine or metalloproteinase activity could be detected . Proteinase activity was higher in culture medium containing BSA or wheat germ extract, as compared to minimal medium . A proportion of the enzyme activity remained in the extracellular glucan sheath . AP was also the only type of proteinase activity in fluid obtained from B . cinerea-infected tissue of apple, pepper, tomato and zucchini . Five B . cinerea genes encoding an AP were cloned and denoted Bcap1-5 . Features of the encoded proteins are discussed . BcAP1, especially, has novel characteristics . A phylogenetic analysis was performed comprising sequences originating from different kingdoms . BcAP1 and BcAP5 did not cluster in a bootstrap-supported clade . BcAP2 clusters with vacuolar APs . BcAP3 and BcAP4 cluster with secreted APs in a clade that also contains glycosylphosphatidylinositol-anchored proteinases from Saccharomyces cerevisiae and Candida albicans . All five Bcap genes are expressed in liquid cultures . Transcript levels of Bcap1, Bcap2, Bcap3 and Bcap4 are subject to glucose and peptone repression . Transcripts from all five Bcap genes were detected in infected plant tissue, indicating that at least part of the AP activity in planta originates from the pathogen. Microbiology, 2004 Jul, 150(Pt 7), 2415 - 28 ALS3 and ALS8 represent a single locus that encodes a Candida albicans adhesin; functional comparisons between Als3p and Als1p; Zhao X et al.; The ALS (agglutinin-like sequence) gene family of Candida albicans encodes eight cell-surface glycoproteins, some of which are involved in adherence to host surfaces . A mutational analysis of each ALS gene is currently being performed to deduce the functions of the encoded proteins and to better understand the role of these proteins in C . albicans biology and pathogenesis . This paper describes construction of an als3/als3 mutant and comparison of its phenotype to an als1/als1 strain . Efforts to disrupt ALS3 indicated that the gene could be deleted in two transformation steps, suggesting that the gene is encoded by a single locus and that the ALS3-like locus, ALS8, does not exist . Strains lacking ALS3 or ALS1 did not exhibit a defect in germ tube formation when grown in RPMI 1640 medium, but the als1/als1 mutant formed significantly fewer germ tubes in Lee medium . Analysis of ALS3 and ALS1 promoter activity using green fluorescent protein (GFP) reporter strains and flow cytometry showed that when cells are placed into medium that promotes germ tube formation, ALS1 is transcribed prior to ALS3 . Comparison of the mutant strains in adhesion assays showed that the als3/als3 strain was defective in adhesion to both human umbilical vein endothelial cells (HUVEC) and buccal epithelial cells (BEC), but not to fibronectin-coated plastic plates . In contrast, the als1/als1 strain showed decreased adherence to HUVEC, but adherence to BEC and fibronectin were the same as wild-type controls . Inoculation of the buccal reconstituted human epithelium (RHE) model of oral candidiasis with the mutant strains showed nearly a total lack of adhesion and epithelial destruction by the als3/als3 mutant while the als1/als1 strain showed only a slightly reduced degree of epithelial destruction compared to the wild-type control . Adhesion data presented here suggest that, in the assays performed, loss of Als3p affects C . albicans adhesion more than loss of Als1p . Collectively, these results demonstrate functional similarities and differences between Als1p and Als3p, and suggest the potential for more complex interrelationships between the ALS genes and their encoded proteins. Microbiology, 2004 Jul, 150(Pt 7), 2197 - 208 The CaCTR1 gene is required for high-affinity iron uptake and is transcriptionally controlled by a copper-sensing transactivator encoded by CaMAC1; Marvin ME et al.; The ability of Candida albicans to acquire iron from the hostile environment of the host is known to be necessary for virulence and appears to be achieved using a similar system to that described for Saccharomyces cerevisiae . In S . cerevisiae, high-affinity iron uptake is dependent upon the acquisition of copper . The authors have previously identified a C . albicans gene (CaCTR1) that encodes a copper transporter . Deletion of this gene results in a mutant strain that grows predominantly as pseudohyphae and displays aberrant morphology in low-copper conditions . This paper demonstrates that invasive growth by C . albicans is induced by low-copper conditions and that this is augmented in a Cactr1-null strain . It also shows that deletion of CaCTR1 results in defective iron uptake . In S . cerevisiae, genes that facilitate high-affinity copper uptake are controlled by a copper-sensing transactivator, ScMac1p . The authors have now identified a C . albicans gene (CaMAC1) that encodes a copper-sensing transactivator . A Camac1-null mutant displays phenotypes similar to those of a Cactr1-null mutant and has no detectable CaCTR1 transcripts in low-copper conditions . It is proposed that high-affinity copper uptake by C . albicans is necessary for reductive iron uptake and is transcriptionally controlled by CaMac1p in a similar manner to that in S . cerevisiae. J Biol Chem, 2004 Sep 24, 279(39), 40852 - 60 Epub 2004 Jul 12. The ATC1 gene encodes a cell wall-linked acid trehalase required for growth on trehalose in Candida albicans; Pedreno Y et al.; After screening a Candida albicans genome data base, the product of an open reading frame (IPF 19760/CA2574) with 41% identity to Saccharomyces cerevisiae vacuolar acid trehalase (Ath1p) was identified and named Atc1p . The deduced amino acid sequence shows that Atc1p contains an N-terminal hydrophobic signal peptide and 20 potential sites for N-glycosylation . C . albicans homozygous mutants that lack acid trehalase activity were constructed by gene disruption at the two ATC chromosomal alleles . Analysis of these null mutants shows that Atc1p is localized in the cell wall and is required for growth on trehalose as a carbon source . An Atc1p endowed with acid trehalase activity was obtained by an in vtro transcription-translation coupled system . These results strongly suggest that ATC1 is the structural gene encoding cell wall acid trehalase in C . albicans . Determinations of ATC1 mRNA expression as well as acid trehalase activity in the presence and absence of glucose point out that ATC1 gene is regulated by glucose repression . Clin Infect Dis, 2004 Jul 15, 39 Suppl 1, S11 - 4 Current antimicrobial usage for the management of infections in leukemic patients in Japan: results of a survey; Yoshida M et al.; We report the findings of a questionnaire distributed by the Committee of Supportive Care of the Japan Adult Leukemia Study Group to 196 hospitals throughout Japan . For antimicrobial prophylaxis, the oral quinolones are prescribed by 38% of physicians and polymixin B by 31% . For antifungal prophylaxis, amphotericin B is prescribed by 42% of physicians and fluconazole by 41% . Febrile neutropenia is empirically treated with cephalosporin or carbapenem monotherapy by 35% of physicians . Overall, dual therapy (i.e., an aminoglycoside plus a cephalosporin, a carbapenem, or an antipseudomonal penicillin) is prescribed by 50% of physicians . When response to initial empirical therapy does not occur after 3-4 days, 51% of physicians add an antifungal agent; fluconazole is preferred to amphotericin B (prescribed by 66% vs . 28% of physicians) . For the treatment of fungemia due to Candida albicans, fluconazole was prescribed by 59% of physicians in cases of stable disease and amphotericin B was prescribed by 57% of physicians in cases of unstable disease . Amphotericin B is selected to treat invasive aspergillosis, but a dose of 0.5-0.7 mg/kg, inadequate for this disease, is prescribed by 44% of physicians . Granulocyte colony-stimulating factor is prescribed to treat patients with acute myelogenous leukemia who have life-threatening infections (27% of physicians) or who have clinically or microbiologically documented infections (26% of physicians). Mol Genet Genomics, 2004 Aug, 272(1), 88 - 97 Epub 2004 Jul 13. The white-phase-specific gene WH11 is not required for white-opaque switching in Candida albicans; Park YN et al.; Phenotypic switching between white and opaque cells is important for adaptation to different host environments and for mating in the opportunistic fungal pathogen Candida albicans . Genes that are specifically activated in one of the two cell types are likely to be important for their phenotypic characteristics . The WH11 gene is a white-phase-specific gene that has been suggested to be involved in the maintenance of the white-phase phenotype . To elucidate the role of WH11 in white-opaque switching, we constructed mutants of the C . albicans strain WO-1 in which the WH11 gene was deleted . The Delta wh11 mutants were still able to form both white and opaque cells whose cellular and colony phenotypes were indistinguishable from those of the wild type . Deletion of WH11 also did not affect the activation and deactivation of the white-phase-specific WH11 promoter and the opaque-phase-specific OP4 and SAP1 promoters in the appropriate cell type . Finally, switching from the white to the opaque phase and vice versa occurred with the same frequency in wild-type and Delta wh11 mutants . Therefore, the WH11 gene is not required for phenotypic switching, and its protein product seems to have other roles in white cells, which are dispensable after the switch to the opaque phase. Int Microbiol, 2004 Jun, 7(2), 105 - 12 Dynamics of CaCdc10, a septin of Candida albicans, in living cells and during infection; Gonzalez-Novo A et al.; The morphogenetic program in the pathogenic fungus Candida albicans, including the dimorphic transition, is an interesting field of study, not only because it is absent in the commonly used model yeast Saccharomyces cerevisiae, but because of the close relationship between hyphal development and virulence of C . albicans . We studied one of the most important aspects of fungal morphogenesis--the septin ring--in C . albicans . By using a fusion construct to green fluorescent protein (GFP), the subcellular localization and dynamics of C . albicans Cdc10 in the different morphologies that this fungus is able to adopt was identified . The localization features reached were contrasted and compared with the results obtained from Candida cells directly extracted from an animal infection model under environmental conditions as similar as possible to the physiological conditions encountered by C . albicans during host infection. Bioorg Med Chem, 2004 Aug 1, 12(15), 3971 - 6 (E)- and (Z)-1,2,4-triazolylchromanone oxime ethers as conformationally constrained antifungals; Emami S et al.; A series of 1,2,4-triazolylchromanone oxime ethers were synthesized and tested for in vitro antifungal activity . Many of these derivatives exhibit high activity against Candida albicans, Saccharomyces cerevisiae, Aspergillus niger and Microsporum gypseum. J Infect Dis, 2004 Aug 1, 190(3), 632 - 40 Epub 2004 Jun 28. Role of mannose-binding lectin in the innate defense against Candida albicans: enhancement of complement activation, but lack of opsonic function, in phagocytosis by human dendritic cells; Ip WK et al.; Mannose-binding lectin (MBL) is a serum collectin believed to be of importance in innate immunity . We have investigated the role of MBL in the first-line defense against Candida albicans, an opportunistic fungal pathogen . MBL bound C . albicans via its lectin domain, resulting in agglutination of the organisms upon their outgrowth of hyphae . In a human in vitro MBL system, deposition of C4 fragments on C . albicans was increased when exogenous MBL was added to serum samples from MBL-deficient individuals . Similar enhancement of deposition of iC3b also was observed . MBL and enhanced opsonic C3 fragments mediated by MBL did not facilitate opsonophagocytosis of the organisms by monocyte-derived dendritic cells (DCs) . However, MBL was found to inhibit the growth of C . albicans independently of complement activation, although, with complement activation, further inhibition was observed . We concluded that MBL plays an important role in the first-line defense against C . albicans without the need for opsonophagocytosis by DCs, in which a direct interaction of MBL with C . albicans results in agglutination and accelerated complement activation via the lectin pathway, leading to inhibition of growth. J Infect Dis, 2004 Aug 1, 190(3), 624 - 31 Epub 2004 Jun 22. Requirement of interleukin-17A for systemic anti-Candida albicans host defense in mice; Huang W et al.; T cells are required for normal host defense against fungal infection, and individuals with T cell-deficiency syndromes are highly susceptible to fungal pathogens . Interleukin (IL)-17A is a proinflammatory cytokine that interconnects myeloid and lymphoid host defense . The role of murine (m) IL-17A/mIL-17A receptor (R) interactions was evaluated in a murine model of systemic candidiasis . In response to systemic challenge with Candida albicans, expression of mIL-17A was induced, and IL-17AR knockout (IL-17AR(-/-)) mice had dose-dependent, substantially reduced survival . Fungal burden in the kidneys of IL-17AR(-/-) mice was dramatically increased (25-fold at 96 h) . In IL-17AR(-/-) mice, both mobilization of peripheral neutrophils and their influx to infected organs were significantly impaired and delayed . In vivo expression of mIL-17A protected normal mice from a lethal dose of C . albicans (100% at day 7 and 65% at day 42) . The data suggest that the mIL-17A/mIL-17AR system is required for normal fungal host defense in vivo . IL-17A could have potential as a therapeutic cytokine for systemic C . albicans infections in immunocompromised patients with cancer or advanced acquired immunodeficiency syndrome. J Infect Dis, 2004 Aug 1, 190(3), 605 - 12 Epub 2004 Jun 25. Tissue-associated cytokine expression in HIV-positive persons with oropharyngeal candidiasis; Lilly EA et al.; Oropharyngeal candidiasis (OPC), caused by Candida albicans, is the most common infection in human immunodeficiency virus (HIV)-positive persons . Although CD4(+) T cells are considered to be important for host defense against C . albicans at the oral mucosa, a recent immunohistochemical evaluation of T cells in OPC lesions of HIV-positive persons with reduced CD4(+) T cells showed high numbers of CD8(+) T cells . The present study investigated tissue-associated expression of cytokine and chemokine mRNA at the site of infection . Results showed some effects of HIV (primarily increased chemokine mRNA levels) but little effect of blood CD4(+) T cells . In contrast, mRNA for several proinflammatory, T helper, and CD8(+) T cell-associated cytokines and chemokines were increased in subjects with OPC versus those without . These results support the presence of CD8(+) T cells in OPC lesions and suggest evidence for a response against OPC, despite reduced levels of CD4(+) T cells. J Clin Microbiol, 2004 Jul, 42(7), 3117 - 9 Further standardization of broth microdilution methodology for in vitro susceptibility testing of caspofungin against Candida species by use of an international collection of more than 3,000 clinical isolates; Pfaller MA et al.; The influence of test variables on in vitro susceptibility testing of caspofungin was examined with 694 isolates of Candida albicans including seven laboratory-derived glucan synthesis mutants . The conditions providing the greatest separation between the mutant strains and the clinical isolates were RPMI medium, MIC end point criterion of partial inhibition, and incubation for 24 h . These testing conditions were then applied to 3,322 isolates of Candida spp . (3,314 clinical isolates and eight glucan synthesis mutants) . Among the 11 isolates for which caspofungin MICs were >/=2 microg/ml, eight were accounted for by the glucan synthesis mutants . The MICs for >99% of isolates were </=1 microg/ml, and thus these isolates were differentiated from strains with reduced in vitro and in vivo susceptibilities to caspofungin. Jpn J Thorac Cardiovasc Surg, 2004 Jun, 52(6), 318 - 21 Prosthetic valve endocarditis due to Candida albicans treated successfully with medical treatment alone; Noguchi M et al.; Prosthetic valve endocarditis (PVE) caused by Candida species is associated with high morbidity and mortality . A combination of surgical resection and antifungal drug therapy is the golden standard for treatment, yet surgical intervention is not possible in all cases of Candida PVE . We report a case of PVE due to Candida albicans cured by medical treatment alone . This case suggests that, in some instances, Candida PVE can be managed medically with antifungal therapy . Such a conservative approach should be applied with caution and necessitates very close follow-up on a long-term basis. Genome Biol . 2004;5(7):230 . Epub 2004 Jun 11. Candida albicans genome sequence: a platform for genomics in the absence of genetics; Odds FC et al.; Publication of the complete diploid genome sequence of the yeast Candida albicans will accelerate research into the pathogenesis of Candida infections . Comparative genomic analysis highlights genes that may contribute to C . albicans survival and its fitness as a human commensal and pathogen. Biomedica, 2004 Mar, 24(1), 7 - 12 {The 'bull's eye' pattern in hepatic tomography}; Cortes JA et al.; Invasive fungal infections are more commonly found in patients who develop neutropenia after chemotherapy . A 4-year-old girl with diagnosis of acute lymphoid leukemia developed febrile neutropenia after chemotherapy . Broad spectrum antibiotics and antimycotic therapy were initiated . Candida albicans was isolated and Entamoeba histolytica was observed in stool examination . Chronic disseminated candidiasis had developed and was treated with amphotericin B, initially, and fluconazol . Computed tomography images were obtained that demonstrated a classic 'bull's eye' pattern; a concurrent histological study confirmed the diagnosis . Candida spp . is the major cause of opportunistic mycosis in immunosuppresed patients receiving chemotherapy for haematologic malignancies . An initial infection results in disseminated candidiasis, which persists and becomes chronic . In the 4-year-old patient, the identified risk factors consisted of a previous therapy with broad spectrum antibiotics, the gastrointestinal tract colonization with Candida albicans and prolonged neutropenia . Imaging diagnoses are made by ultrasonography, computed tomography and magnetic resonance . With ultrasound and tomography, 4 distinct patterns have been described . Pattern 1 ('wheels within wheels') and 2 ('bull's eye') are important, since they are characteristic of chronic disseminated candidiasis . The third pattern (hypoechoic image) is the most common finding with both techniques . In the current patient, patterns 2 and 3 were seen and the diagnosis was confirmed by histological study. World J Gastroenterol, 2004 Jul 15, 10(14), 2124 - 6 Intestinal colonization with Candida albicans and mucosal immunity; Bai XD et al.; AIM: To observe the relationship between intestinal lumen colonization with Candida albicans and mucosal secretory IgA (sIgA) . METHODS: A total of 82 specific-pathogen-free mice were divided randomly into control and colonization groups . After Candida albicans were inoculated into specific-pathogen-free mice, the number of Candida albicans adhering to cecum and mucosal membrane was counted . The lymphocyte proliferation in Peyer's patch and in lamina propria was shown by BrdU incorporation, while mucosal sIgA (surface membrane) isotype switch in Peyer's patch was investigated . IgA plasma cells in lamina propria were observed by immunohistochemical staining . Specific IgA antibodies to Candida albicans were measured with ELISA . RESULTS: From d 3 to d 14 after Candida albicans gavaging to mice, the number of Candida albicans colonizing in lumen and adhering to mucosal membrane was sharply reduced . Candida albicans translocation to mesenteric lymph nodes occurred at early time points following gavage administration and disappeared at later time points . Meanwhile, the content of specific IgA was increased obviously . Proliferation and differentiation of lymphocytes in lamina propria were also increased . CONCLUSION: Lymphocytes in lamina propria play an important role in intestinal mucosal immunity of specific-pathogen-free mice when they are first inoculated with Candida albicans . The decreasing number of Candida albicans in intestine is related to the increased level of specific IgA antibodies in the intestinal mucus. Int Immunol, 2004 Aug, 16(8), 1125 - 31 Epub 2004 Jul 05. Antiarthritic effect of VIP in relation to the host resistance against Candida albicans infection; Zafirova Y et al.; Vasoactive intestinal peptide (VIP) is one of the prospective candidates for clinical application in r