Candida albicans

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 rheumatoid arthritis (RA) . Its antiarthritic effect is associated with the suppression of inflammatory and autoimmune responses . The ability of VIP to trigger a shift towards Th2 immunity suggests that anti-infectious host resistance might be affected . In the present study VIP was applied at the initiation and at the established phase of collagen-induced arthritis (CIA) . Mice developed Th2 dominant anti-collagen response . The susceptibility to primary and secondary Candida albicans infection was determined after VIP administration at the established CIA . The percentage of survivors, kidney colonization, cytokine secretion by splenocytes and specific antibody synthesis were assessed . Reduced TNF-alpha production but not IFN-gamma and IL-10 was observed after the first challenge with the pathogen in CIA mice treated with VIP while the percentage of survivors was not significantly changed . The adaptive immune response was impaired in VIP-treated mice as they were more susceptible to reinfection, showed increased kidney colonization and suppressed anti-Candida IgG antibody production.

Indian J Exp Biol, 2004 Mar, 42(3), 326 - 9
Evaluation of phytochemical and antimicrobial properties of leaf extract of Tapinanthus sessilifolius (P . Beauv) van Tiegh; Tarfa FD et al.; Leaf extracts of T . sessilifolius growing on five different host plants (Psidium guajava, Citrus lemon, Vernonia amygdalina, Persea americana and Jatropa curcas) were evaluated for antimicrobial activity of the plant . Powdered leaves of T . sessilifolius collected from each host plant was divided into two portions . One portion was used for aqueous infusion and the other portion was successively extracted with hexane, ethylacetate and methanol . Infusion of aqueous extract of powdered leaves did not show antimicrobial effect even at the concentration of 1000 and 2000 microg/ml on test microorganisms (Staph . aureus, E . coli, Bacillus subtilis, Pseudomonas aeruginosa and Candida albicans) . However in broth culture, methanolic and hexane extract had MIC range of 62.5-500 microg/ml and ethylacetate extract had 250-500 microg/ml . Phytochemical screening of leaf samples of T . sessilifolius collected from different host plants showed positive test for hydrolysable tannins, saponins, flavonoids, terpenes, cardiac glycoside, reducing sugars and proteins . LD50 concentration was found to be > 1.500 mg/kg for samples from P . guajava; 489.89 mg/kg for J . curcas and C . lemon; and 692 mg/kg for V . amydalina in mice.

Med Sci Monit, 2004 Jul, 10(7), BR198 - 206 Epub 2004 Jun 29.
Candida albicans switch phenotypes display differential levels of fitness; Vargas K et al.; BACKGROUND: Candidiasis is a significant cause of morbidity and mortality in immunosuppressed individuals . Currently, there are limited treatment options for this opportunistic infection and there have been many reports in the literature of increased resistance to treatment . There is evidence to suggest that phenotypic switching of Candida albicans may play a role in this resistance . It was therefore, the purpose of this study to assess the variability among C . albicans switch phenotypes isolated from a single strain . MATERIAL/METHODS: Four switch phenotypes from a single strain of Candida albicans isolated from an HIV+ individual were evaluated for growth rates, post-antifungal effects in the presence of fluconazole, fluconazole uptake and morphological changes after fluconazole exposure . RESULTS: We found that, overall, smooth white and very wrinkled grew significantly faster then either ring or heavily myceliated and were most drastically affected by 4X and 6X MIC concentrations of fluconazole over a 24 hour period . When all four phenotypes were exposed to 64 . CONCLUSIONS: These results suggest that switch phenotypes from Candida albicans serve different roles in providing adaptability and survivability under differing conditions.

Int J Dermatol, 2004 Jul, 43(7), 498 - 502
Onychomycosis in central India: a clinicoetiologic correlation; Garg A et al.; BACKGROUND: Onychomycosis is mainly caused by dermatophytes, but yeasts and nondermatophyte molds have also been implicated, giving rise to diverse clinical presentations . The etiological agents of the disease may show geographic variation . The aim of the present study was to isolate the causative pathogens and to determine the various clinical patterns of onychomycosis in central India . METHODS: The study population comprised 90 patients with onychomycosis . Nail samples were collected for direct microscopic examination and culture . Clinical patterns were noted and correlated with causative pathogens . RESULTS: The male : female ratio was 3:1 and the mean age was 29.40 +/- 13.61 years . Fingernails were involved in 60%, toenails in 26.67% and both fingernails and toenails in 13.34% of the 90 patients . The clinical types noted were distolateral subungual onychomycosis (64.44%), total dystrophic onychomycosis (17.78%), proximal subungual onychomycosis with paronychia (12.2%), proximal subungual onychomycosis without paronychia (4.44%) and superficial white onychomycosis (1.11%) . Dermatophytes were the most common pathogens isolated, being found in 24 patients (26.36%) {Tricophyton rubrum (23.07%), Tricophyton verrucosum (2.22%) and Epidermophyton floccosum (1.11%)}, followed by Candida albicans, which was found in 22 patients (24.27%) . Thirty-six (39.58%) nondermatophyte molds were isolated from 29 patients . Of these 29 cases, six were associated with Tricophyton rubrum, which was considered the primary pathogen . CONCLUSIONS: Distolateral subungual onychomycosis was the most common clinical presentation; however, total dystrophic onychomycosis and proximal subungual onychomycosis were not uncommon in this part of India . Tricophyton rubrum and Candida albicans were the major pathogens . The clinicoetiologic correlation revealed that a single pathogen could give rise to more than one clinical type.

Planta Med, 2004 Jun, 70(6), 531 - 5
Photo-activated DNA binding and antimicrobial activities of furoquinoline and pyranoquinolone alkaloids from rutaceae; Hanawa F et al.; To find novel photo-active compounds of potential use in photochemotherapy from higher plants, photo-activated antimicrobial and DNA binding activities of the furoquinolines, skimmianine, kokusaginine, and haplopine, and a pyranoquinolone, flindersine, from two species of Rutaceae plants were investigated . TLC overlay assays against a methichillin-resistant strain of Staphylococcus aureus and Candida albicans were employed to test antimicrobial properties . All of the tested compounds showed photo-activated antimicrobial activity against S . aureus in the order of kokusaginine > haplopine, flindersine > skimmianine . Weaker activity was found for C . albicans . Photo-activated DNA binding activity of these compounds was investigated by a method using restriction enzymes and a specially designed 1.5 kb DNA fragment . Kokusaginine showed inhibition against all of the 16 restriction enzymes . Haplopine showed a similar inhibition pattern but the binding activity against Asc I and Sma I with restriction sequences consisting only of G and C was very weak . Skimmianine showed binding activity against Xba I, BciV I, Sal I, Pst I, Sph I and Hind III, but very weak or no activity was found for the other restriction enzymes . A pyranoquinolone, flindersine, showed no activity against any of the restriction enzymes . Photo-activated DNA binding activity of furoquinolines was therefore in the order of kokusaginine > haplopine > skimmianine, which was the same order as their photo-activated antimicrobial activity against S . aureus . Therefore, it was concluded that DNA is one of the cellular targets for the furoquinolines to exert their biological activities, similar to psoralens . However, because flindersine showed photo-activated antimicrobial activity against S . aureus but did not show photo-activated DNA binding activity, it is clear that there are other cellular target components for this compound to exert photo-toxic activity.

Mol Biol Cell, 2004 Sep, 15(9), 4179 - 90 Epub 2004 Jun 30.
A conserved stress-activated protein kinase regulates a core stress response in the human pathogen Candida albicans; Smith DA et al.; Previous work has implicated the Hog1 stress-activated protein kinase (SAPK) in osmotic and oxidative stress responses in the human pathogen Candida albicans . In this study, we have characterized the role of Hog1 in mediating these and other stress responses in C . albicans . We provide evidence that a SAPK-dependent core stress response exists in this pathogen . The Hog1 SAPK is phosphorylated and it accumulates in the nucleus in response to diverse stress conditions . In addition, we have identified Hog1-regulated genes that are induced in response to stress conditions that activate Hog1 . These analyses reveal both activator and repressor functions for the Hog1 SAPK . Our results also demonstrate that stress cross-protection, a classical hallmark of the core stress response, occurs in C . albicans between stresses that activate the Hog1 SAPK . Importantly, we find that the core stress response in C . albicans has adapted to the environmental niche of this human pathogen . This niche specificity is reflected by the specific environmental conditions that drive the Hog1-regulated core stress response in C . albicans and by differences in the molecular circuitry that control this response.

Bioorg Med Chem Lett, 2004 Aug 2, 14(15), 3975 - 8
Rhodanine-3-acetic acid derivatives as inhibitors of fungal protein mannosyl transferase 1 (PMT1); Orchard MG et al.; The first inhibitors of fungal protein: mannosyl transferase 1 (PMT1) are described . They are based upon rhodanine-3-acetic acid and several compounds have been identified, for example, 5-{{3-(1-phenylethoxy)-4-(2-phenylethoxy)phenyl}methylene}-4-oxo-2-thioxo-3-thiazolidineacetic acid (5a), which inhibit Candida albicans PMT1 with IC(50)s in the range 0.2-0.5 microM . Members of the series are effective in inducing changes in morphology of C . albicans in vitro that have previously been associated with loss of the transferase activity . These compounds could serve as useful tools for studying the effects of protein O-mannosylation and its relevance in the search for novel antifungal agents.

Med Trop (Mars), 2004, 64(1), 39 - 42
{Prevalence and risk factors for otomycosis treated in the hospital setting in Abidjan (Ivory Coast)}; Yavo W et al.; Otomycosis is frequently encountered in tropical and subtropical zones . In Ivory Coast diagnosis of this disease is often based solely on the clinical symptoms . The objectives of this study were to determine the prevalence, predisposing factors, and etiologic agents associated with otomycosis at the Treichville University Hospital Center in Abidjan, Ivory Coast . Mycological examinations were performed on specimens obtained from 115 patients presenting with external otitis at the d'Oto-Rhino-Laryngology Department . Fungi-positive cultures were obtained in 49 patients for an overall otomycosis prevalence of 42.6 (95% Confidence Interval (CI), 34.4-52.2) . Univariate analysis showed that the predisposing factors for otomycosis were frequent swimming in natural or artificial pools (Relative Risk (RR) 3.7; CI 1.7-8.1), daily ear cleaning (RR 3.5; CI 1.8-6.8) and excessive use of eardrops containing antibiotics and corticoids (RR = 9.3; IC95% = 4.3-20.1) . The most common etiologic agents were Aspergillus flavus (20.4%), Candida albicans (16.3%), Candida parapsilosis (14.3%), and Aspergillus niger (12.2%) . A combination of two agents was found in five cases . These data show that otomycosis is endemic in Ivory Coast . Management of otomycosis must include mycological examination for diagnosis as well as changing behavior patterns leading to infection.

Mediators Inflamm, 2004 Jun, 13(3), 189 - 93
Production of anti-Candida antibodies in mice with gut colonization of Candida albicans; Tansho S et al.; BACKGROUND: Production of antibodies that are specific for allergens is an important pathological process in inflammatory allergic diseases . These contain the antibodies against antigens of Candida albicans, one of the normal microbial flora in an intestinal tract . We studied the effects of the prednisolone administration on the production of anti-Candida antibodies in the gastrointestinally C . albicans-colonized mice . METHODS AND MATERIALS: BALB/c mice, treated with antibacterial antibiotics to decontaminate indigenous intestinal bacterial flora, were inoculated intragastrically with C . albicans . The mice, in which C . albicans grows intestinally, were administered prednisolone to induce temporary immunosuppression . The Candida growth in their intestinal tract and their antibody response to Candida were examined . RESULTS: Antibiotic treatment allowed establishment of C . albicans gastrointestinal colonization, but did not cause subsequent systemic dissemination of C . albicans in all the animals . When these animals received an additional treatment with prednisolone, they showed a significantly higher population of C . albicans in their feces than those of animals treated with antibiotics alone, and the organisms were recovered even from their kidney . This systemic dissemination by C . albicans appeared to be temporal, because all the mice survived without any symptoms for more than 2 months . Examination of the serum titers of total immunoglobulin (Ig)E antibodies and specific IgE and IgG antibodies against Candida antigens demonstrated that titers of total IgE increased, partially by day 14 and clearly at day 27, in prednisolone-treated Candida-colonized mice . Without prednisolone treatment, an increment of the serum titer was scarcely observed . By day 27, corresponding to the increase of total IgE, the anti-Candida IgE and IgG titer increased in mice of the prednisolone-treated group . CONCLUSION: Administration of prednisolone to Candida-colonized mice can induce production of the IgG, IgE antibodies against Candida antigens, perhaps through temporal systemic dissemination of Candida from the intestinal tract.

Trends Microbiol, 2004 Jul, 12(7), 317 - 24
The distinct morphogenic states of Candida albicans; Sudbery P et al.; The human fungal pathogen, Candida albicans can grow in at least three different morphologies: yeast, pseudohyphae and hyphae . Further morphological forms exist during colony switching, for example, opaque phase cells are oblong, rather than the oval shape of yeast cells . Pseudohyphae and hyphae are both elongated and sometimes there has been little attempt to distinguish between them, as both are "filamentous forms" of the fungus . We review here the differences between them that suggest that they are distinct morphological states . We argue that studies on "filamentous forms" should always include a formal analysis to determine whether the cells are hyphae or pseudohyphae and we suggest some simple experimental criteria that can be applied to achieve this.

Biochemistry, 2004 Jul 6, 43(26), 8483 - 93
Effect of membrane perturbants on the activity and phase distribution of inositol phosphorylceramide synthase; development of a novel assay; Aeed PA et al.; The effect of 26 different membrane-perturbing agents on the activity and phase distribution of inositol phosphorylceramide synthase (IPC synthase) activity in crude Candida albicans membranes was investigated . The nonionic detergents Triton X-100, Nonidet P-40, Brij, Tween, and octylglucoside all inactivated the enzyme . However, at moderate concentrations, the activity of the Triton X-100- and octylglucoside-solubilized material could be partially restored by inclusion of 5 mM phosphatidylinositol (PI) in the solubilization buffer . The apparent molecular mass of IPC synthase activity solubilized in 2% Triton X-100 was between 1.5 x 10(6) and 20 x 10(6) Da, while under identical conditions, octylglucoside-solubilized activity remained associated with large presumably membrane-like structures . Increased detergent concentrations produced more drastic losses of enzymatic activity . The zwitterionic detergents Empigen BB, N-dodecyl-N,N-(dimethylammonio)butyrate (DDMAB), Zwittergent 3-10, and amidosulfobetaine (ASB)-16 all appeared capable of solubilizing IPC synthase . However, these agents also inactivated the enzyme essentially irreversibly . Solubilization with lysophospholipids again resulted in drastic losses of enzymatic activity that were not restored by the inclusion of PI . Lysophosphatidylinositol also appeared to compete, to some extent, with the donor substrate phosphatidylinositol . The sterol-containing agent digitonin completely inactivated IPC synthase . By contrast, sterol-based detergents such as 3-{(3-cholamidopropyl)dimethylammonio}-1-propanesulfonate (CHAPS), 3-{(3-cholamidopropyl)dimethylammonio}-2-hydroxy-1-propanesulfonate (CHAPSO), and taurodeoxycholate (tDOC) had little or no effect on the enzyme activity . The IPC synthase activity in C . albicans membranes remained largely intact and sedimentable at CHAPS concentrations (4%) where >90% of the phospholipids and 60% of the total proteins were extracted from the membranes . At 2.5% CHAPS, a concentration where approximately 50% of the protein and 80% of the phospholipids are solubilized, there was no detectable loss of enzyme activity, and it was found that the detergent-treated membranes had significantly improved properties compared to crude, untreated membranes as the source of IPC synthase activity . In contrast to assays utilizing intact membranes or Triton X-100 extracts, assays using CHAPS- or tDOC-washed membranes were found to be reproducible, completely dependent on added acceptor substrate (C(6)-7-nitro-2-1,3-benzoxadiazol-4-yl (NBD)-ceramide), and >95% dependent on added donor substrate (PI) . Product formation was linear with respect to both enzyme concentration and time, and transfer efficiency was improved more than 20-fold as compared to assays using crude membranes . Determination of kinetic parameters for the two IPC synthase substrates using CHAPS-washed membranes resulted in K(m) values of 3.3 and 138.0 microM for C(6)-NBD-ceramide and PI, respectively . In addition, the donor substrate, PI, was found to be inhibitory at high concentrations with an apparent K(i) of 588.2 microM.

J Ind Microbiol Biotechnol, 2004 Jul, 31(6), 255 - 60 Epub 2004 Jun 22.
Colonization of hydrophilic contact lenses by yeast; Marques-Calvo MS; The growth of six strains of yeast was analyzed in vitro in order to assess their capacity for colonizing (adhesion and invasion) hydrophilic contact lenses . Lenses with different water content were cultured in two culture media for 3, 7, 14, and 21 days . Only strain 93150 of Candida albicans could adhere to and invade the polymers . Specifically, fungal growth was observed in cultures with Sabouraud's broth . The degree of yeast colonization of contact lenses was significantly related to the species, the strain, and the culture medium in which the yeast and lenses were cultured . The results here obtained were compared with those reported for the filamentous fungus Aspergillus niger 2700 . For both microorganisms, the strain and the medium in which the lenses and microorganism were cultured influenced the colonization, but the percentage of colonized lenses, the degree of colonization, and the density and size of the internalized colonies were always noticeably lower for C . albicans 93150 . Colonization by A . niger 2700 was also related to the type of material of the lenses and the incubation period . For both microorganisms, when the strain is right and the growth and development are correct, colonization of hydrophilic contact lenses occurs.

Fungal Genet Biol, 2004 Aug, 41(8), 783 - 93
Candida albicans CTN gene family is induced during macrophage infection: homology, disruption and phenotypic analysis of CTN3 gene; Prigneau O et al.; We have isolated a Candida albicans gene, coding for a putative peroxisomal carnitine acetyl transferase (CTN) protein, which is up-regulated during macrophage infection . In the present study, we describe the disruption of CTN3 gene (previously called CAT3) to gain insight into its potential role during infection . The ability of disrupted Candida mutants to filament was affected by several solid media . Northern blot analysis revealed that CTN3 gene may be involved not only in conditions of cell starvation but also during the process of germination . In agreement with the putative peroxisomal localization of the corresponding protein, we observed a strong glucose repression of CTN3 gene and, on the contrary, high level of transcription by carbon sources that induce the formation of peroxisomal proteins . Furthermore, we showed the existence of two additional C . albicans CTN encoding sequences, which are also induced during macrophage infection.

Int Dent J, 2004 Jun, 54(3), 154 - 8
Clinical and microbiological efficacy of chlorine dioxide in the management of chronic atrophic candidiasis: an open study; Mohammad AR et al.; OBJECTIVE: To assess the clinical and microbiological efficacy of chlorine dioxide (ClO2) as a topical antiseptic for the treatment of chronic atrophic candidiasis in geriatric patients . PARTICIPANTS: Thirty patients with chronic atrophic candidiasis . METHODS: Patients were instructed to rinse the mouth with 0.8% ClO2 mouth rinse (DioxiDent) twice daily for one minute and to soak their dentures overnight in the ClO2 for 10 days . Patients were evaluated both clinically and microbiologically at baseline and after 10 days, and any significant side effects were recorded . The clinical appearance of the oral soft tissues was scored on a scale of 0-3 (0 indicating no clinical signs, 1 indicating involvement of < 25% of the palatal mucosa, 2 indicating involvement of 25-50% of the palatal mucosa, and 3 indicating marked erythema involving > 50% of the palatal mucosa) . Microbiological testing was undertaken to determine the number of colony forming units (CFUs) of Candida albicans . RESULTS: ClO2 significantly improved the clinical appearance and microbial count (p < 0.001) after treatment, without significant side effects . Results showed marked improvement in the clinical appearance of the tissues after 10 days, with total resolution in the majority of cases . The total CFU/ml ranged from 15,000-53,000 at baseline and was reduced to < or = 500 after 10 days of treatment (p < 0.001) . The mean clinical score was 2.50 at baseline, and was reduced to 0.17 after 10 days of treatment (p < 0.001) . CONCLUSIONS: Within the limitations of this pilot study, the effectiveness of topical chlorine dioxide (0.8%) in the management of chronic atrophic candidiasis was demonstrated . ClO2 provided a safe and clinically effective option in the management of chronic atrophic candidiasis.

Mol Biol Cell, 2004 Jul, 15(7), 3167 - 80
APSES proteins regulate morphogenesis and metabolism in Candida albicans; Doedt T et al.; Fungal APSES proteins regulate morphogenetic processes, including filamentation and differentiation . The human fungal pathogen Candida albicans contains two APSES proteins: the regulator Efg1p and its homologue Efh1p, described here . Overexpression of EFG1 or EFH1 led to similar phenotypes, including pseudohypha formation and opaque-white switching . An efh1 deletion generated no phenotype under most conditions but caused hyperfilamentation in an efg1 background under embedded or hypoxic conditions . This suggests cooperation of these APSES proteins in the suppression of an alternative morphogenetic signaling pathway . Genome-wide transcriptional profiling revealed that EFG1 and EFH1 regulate partially overlapping sets of genes associated with filament formation . Unexpectedly, Efg1p not only regulates genes involved in morphogenesis but also strongly influences the expression of metabolic genes, inducing glycolytic genes and repressing genes essential for oxidative metabolism . Using one- and two-hybrid assays, we further demonstrate that Efg1p is a repressor, whereas Efh1p is an activator of gene expression . Overall, the results suggest that Efh1p supports the regulatory functions of the primary regulator, Efg1p, and indicate a dual role for these APSES proteins in the regulation of fungal morphogenesis and metabolism.

J Antibiot (Tokyo), 2004 Apr, 57(4), 253 - 9
FR171456, a novel cholesterol synthesis inhibitor produced by Sporormiella minima No . 15604 . I . Taxonomy, fermentation, isolation, physico-chemical properties; Hatori H et al.; FR171456 and FR173945, novel and potent cholesterol synthesis inhibitors, have been isolated from the fermentation broth of a fungal strain No . 15604 . This strain was identified Sporormiella minima from its mycological characteristics . FR171456 and FR173945 strongly inhibited cholesterol synthesis in human hepatoma cell line Hep G2 . These compounds also have in vitro antifungal activity against Candida albicans and Aspergillus fumigatus.

J Chemother, 2004 Apr, 16(2), 179 - 86
Fenticonazole nitrate for treatment of vulvovaginitis: efficacy, safety, and tolerability of 1-gram ovules, administered as ultra-short 2-day regimen; Fernandez-Alba J et al.; Because of its potential as a low cost first-line monotherapy for the most common vulvovaginal infections, we evaluated fenticonazole nitrate in a prospective, open-label, multicenter pilot study with 101 sexually active women (per-protocol; 16 to 61 years of age) with vulvovaginitis involving single or mixed infections with Candida albicans, Trichomonas vaginalis, and/or Gardnerella vaginalis . Fenticonazole nitrate (1 g) was administered as vaginal ovules, once daily on days 1 and 3 . Eradication (direct phase-contrast microscopy of vaginal swabs and/or microbiological culture) on day 8 was 90% (C . albicans, 26/29, p < 0.001), 70% (T . vaginalis, 7/10, p = 0.161), 67% (G . vaginalis, 22/33, p < 0.009), and 45% (mixed infection, 13/29, p = 0.001) . After 28 days, relapse was 0% for candidiasis and trichomoniasis, 27% (6/22) for G . vaginalis, and 23% (3/13) for mixed infection . Overall, eradication of all offending pathogens was achieved in 67% of the total per-protocol population, with a relapse rate of only 16% . Score sums for symptoms improved from 7.0 (baseline) to 1.7 (day 8), and 0.71 (day 28), (p < 0.001) . Treatment was safe and well tolerated . The results of our pilot study suggest that application of fenticonazole nitrate 1 g intravaginal ovules on 2 alternate days is a suitable first-line treatment of vulvovaginitis with acceptable broad-spectrum efficacy against the most commonly involved pathogens and with a low rate of early relapse, reserving antibiotics for patients with treatment failure or relapse of infection . Our results should encourage further examination of this approach in larger and well controlled clinical trials.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2733 - 5
Proteomic analysis of azole resistance in Candida albicans clinical isolates; Hooshdaran MZ et al.; Changes in protein expression within a matched set of Candida albicans isolates representing the acquisition of azole resistance were examined by two-dimensional polyacrylamide gel electrophoresis and peptide mass fingerprinting . Proteins differentially expressed in association with azole resistance included Grp2p, Ifd1p, Ifd4p, Ifd5p, and Erg10p, a protein involved in the ergosterol biosynthesis pathway.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2730 - 2
Micafungin enhances neutrophil fungicidal functions against Candida pseudohyphae; Gil-Lamaignere C et al.; We evaluated the effect of the combination of micafungin and polymorphonuclear leukocytes (PMN) against hyphae of Candida albicans and Candida dubliniensis . Micafungin enhanced the PMN oxidative burst dose dependently . The combination was synergistic (C . albicans) or additive (C . dubliniensis); when PMN were pretreated with granulocyte-macrophage colony-stimulating factor, the combination was more effective.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2700 - 3
Constitutive activation of the PDR16 promoter in a Candida albicans azole-resistant clinical isolate overexpressing CDR1 and CDR2; De Deken X et al.; Candida albicans azole-resistant clinical isolates overexpressing the CDR1 and CDR2 genes (multidrug transporters) also overexpress the PDR16 gene (phosphatidylinositol transfer protein) . We show here that the PDR16 promoter displays higher transcriptional activity following integration in an azole-resistant isolate than in the matched azole-susceptible one . Thus, the upregulation of PDR16 in the resistant strain results from a mutation acting in trans.

J Biomol Struct Dyn, 2004 Aug, 22(1), 91 - 9
Homology modeling of lanosterol 14alpha-demethylase of Candida albicans and Aspergillus fumigatus and insights into the enzyme-substrate Interactions; Sheng C et al.; The crystal structure of 14alpha-sterol demethylase from Mycobacterium tuberculosis (MT_14DM) provides a good template for modeling the three dimensional structure of lanosterol 14alpha-demethylase, which is the target of azole antifungal agents . Homologous 3D models of lanosterol 14alpha-demethylase from Candida albicans (CA_14DM) and Aspergillus fumigatus (AF_14DM) were built on the basis of the crystal coordinates of MT_14DM in complex with 4-phenylimidazole and fluconazole . The reliability of the two models was assessed by Ramachandran plots, Profile-3D analysis, and by analyzing the consistency of the two models with the experimental data on the P450(14DM) . The overall structures of the resulting CA_14DM model and AF_14DM model are similar to those of the template structures . The two models remain the core structure characteristic for cytochrome P450s and most of the insertions and deletions expose the molecular surface . The structurally and functionally important residues such as the heme binding residues, the residues lining the substrate access channel, and residues in active site were identified from the model . To explore the binding mode of the substrate with the two models, 24(28)-methylene-24,25-dihydrolanosterol was docked into the active site of the two models and hydrophobic interaction and hydrogen-bonding were found to play an important role in substrate recognition and orientation . These results provided a basis for experiments to probe structure-function relationships in the P450(14DM) . Although CA_14DM and AF_14DM shared similar core structural character, the active site of the two models were quite different, thus allowing the rational design of specific inhibitors to the target enzyme and the discovery of novel antifungal agents with broad spectrum .

Isr Med Assoc J, 2004 Jun, 6(6), 342 - 5
Candida albicans colonization of dental plaque in elderly dysphagic patients; Aizen E et al.; BACKGROUND: Dysphagia is a common disorder among the elderly population . As many as 50% of nursing home residents suffer from dysphagia . It is important to identify patients at increased risk for colonization of dental and denture plaque by pathogenic organisms in order to prevent associated disease . OBJECTIVES: To quantify the prevalence and evaluate the effect of dental and denture plaque colonization by Candida albicans in hospitalized elderly dysphagic patients as a complication of stroke, as well as the effect of systemic antimicrobial therapy on C . albicans colonization in these patients . METHODS: We evaluated dysphagia and antibiotic therapy as risk factors for dental and denture plaque colonization by C . albicans in elderly stroke rehabilitating patients with dysphagia, as compared to elderly non-dysphagic stroke and non-stroke rehabilitating patients on days 0, 7 and 14 following admission to the Fliman Geriatric Rehabilitation Hospital . RESULTS: The risk of C . albicans colonization of dental plaque was greater in dysphagic patients than in those without dysphagia on day 0 (50% vs . 21%, P = 0.076), day 7 (58 vs . 15.2%, P = 0.008) and day 14 (58 vs . 15.2%, P = 0.08) . Similarly, patients on antibiotic therapy were at greater risk for C . albicans colonization of dental plaque on day 0 (56 vs . 11%, P = 0.002), day 7 (44 vs . 14.8%, P = 0.04) and day 14 (39 vs . 19%, P = 0.18) . The risk of C . albicans colonization of denture plaque as opposed to dental plaques in non-dysphagic patients was significantly greater on day 0 (45.7 vs . 21.2%, P = 0.03), day 7 (51.4 vs . 15.1%, P = 0.0016) and day 14 (54.3 vs . 15.1%, P = 0.0007) . Dysphagia did not increase the risk of denture plaque colonization by C . albicans . CONCLUSIONS: Both dysphagia and antibiotic therapy are risk factors for C . albicans colonization of dental plaque, and although dysphagia does not significantly increase colonization of denture plaque, denture wearers are at greater risk of such colonization.

FEMS Microbiol Lett, 2004 Jul 1, 236(1), 123 - 8
Partial purification and biochemical characterization of a soluble alpha-glucosidase II-like activity from Candida albicans; Torre-Bouscoulet ME et al.; A soluble alpha-glucosidase was partially purified from Candida albicans cells by a three-step procedure consisting of size-exclusion, ion-exchange and adsorption chromatographies . After the last step, enzyme was enriched about 8.7-fold with a yield of 13% over the starting material and analysis of the purified preparation revealed two major polypeptides of 36 and 47 kDa . The latter was responsible for enzyme activity as visualized with a fluorescent substrate . Nigerose, an alpha-1,3-linked glucose disaccharide, was preferentially hydrolyzed by the purified enzyme over other glucosedisaccharides bearing distinct alpha-linkages . The purified alpha-glucosidase also converted the GlcMan9GlcNAc2 oligosaccharide into the Man9GlcNAc2 product in a time-dependent manner . These and other determined properties are consistent with a type GII alpha-glucosidase probably involved in N-glycan processing.

Oral Microbiol Immunol, 2004 Aug, 19(4), 262 - 9
Role of Candida albicans polymorphism in interactions with oral epithelial cells; Villar CC et al.; BACKGROUND: Candida albicans is a polymorphic organism which undergoes morphologic transition between yeast, pseudohyphal and hyphal forms . The ability of C . albicans to change from yeast to filamentous types is a major virulence determinant of this organism . However, the exact role of hyphal transformation in establishing oral mucosal infection is still poorly understood . METHODS: In this study we used mutants with defects in filamentation, as well as oral strains, which differ in their capacity to form true hyphae, to examine the role of hyphal transformation in the interactions of C . albicans with oral epithelial cells in vitro . These interactions included the ability of these strains to adhere to and injure epithelial cells, as well as their ability to trigger a proinflammatory cytokine response . RESULTS: We found that strains SC5314 and ATCC28366 formed true hyphae on epithelial cells, whereas strain ATCC32077 and the tup1/tup1 mutant formed only pseudohyphae . Double mutant efg1/efg1cph1/cph1 grew exclusively as blastospores . We also found that yeast and pseudohyphal strains showed reduced adherence capacity to oral keratinocytes and caused minimal cell damage . Moreover, we showed that both yeast and pseudohyphal forms have a strongly attenuated proinflammatory phenotype, since they failed to induce significant interleukin (IL)-1alpha and IL-8 responses by oral epithelial cells . CONCLUSIONS: Germination of C . albicans into true hyphae is particularly important in the interactions with oral epithelial cells in vitro.

Mikrobiol Z, 2004 Mar-Apr, 66(2), 35 - 46
{Lytic activity of aerobic sporulating bacteria}; Smirnov VV et al.; The paper deals with lytic activity of 226 strains belonging to 13 species of Bacillus genus, isolated from various econiches, in respect of the cells of Staphylococcus aureus, Escherichia coli and Candida albicans as representatives of Gram-positive, Gram-negative and yeast groups of microorganisms . All the studied strains of bacilli displayed lytic activity of various extent and various spectrum of substrate specificity . Possible dependence of the extent of activity and substrate specificity of bacilli strains on their species belonging and isolation sources did not display distinctly . Two different factors of aggression and antagonism of aerobic bacilli--the capacity to lyse as well as to inhibit growth of test-cultures do not correlate and can either coincide or not coincide . The Bacillus genus and its species are characterized, to a certain extent, as a lytically active group and are estimated as the promising source of obtaining highly active producers of lytic enzymes different as to their properties . Highly active producers of lytic enzymes with the primary degree of lysis from 30% to 60% of the cells of test cultures during 1 hour of the reaction have been obtained.

Bioorg Med Chem Lett, 2004 Jul 16, 14(14), 3821 - 4
Amidine derived 1,3-diazabuta-1,3-dienes as potential antibacterial and antifungal agents; Bedi PM et al.; Several 1-aryl-2-phenyl-4-piperidino-4-thioalkyl-1,3-diazabuta-1,3-dienes were prepared by the treatment of N-arylimino isothiocyanate with piperidine followed by S-alkylation with alkyl iodides in the presence of dry acetone and potassium hydroxide . The constitution of the products was supported by IR, PMR and mass spectral study . The compounds synthesized were tested in in vitro against E . coli, S . aureus, P . aeruginosa, B . cereus and B . subtilis and fungal stains, Candida albicans and Aspergillus niger . Standard drugs were also tested under identical conditions for comparing the results.

J Enzyme Inhib Med Chem, 2004 Feb, 19(1), 85 - 90
Antifungal cobalt(II), copper(II), nickel(II) and zinc(II) complexes of furanyl-,thiophenyl-, pyrrolyl-, salicylyl- and pyridyl-derived cephalexins; Chohan ZH et al.; Some novel cephalexin-derived furanyl, thiophenyl, pyrrolyl, salicylyl and pyridyl Schiff's bases and their cobalt (II), copper (II), nickel (II) and zinc (II) complexes have been synthesized and studied for their antifungal properties against Trichophyton longifusus, Candida albicans, Aspergillus flavus, Microsporum canis, Fusarium solani and Candida glaberata . The presence of metal ions in the investigated Schiff's base complexes reported here lead to significant antifungal activity, whereas the parent ligands were generally less active.

Rev Esp Quimioter, 2004 Mar, 17(1), 48 - 56
{Usefulness of the E-test and its assay conditions in the study of the interaction of antifungal agents . A pilot study}; Canton E et al.; Preliminary data from a pilot study to assess the usefulness of the E-test in the study of antimycoctics are presented, evaluating assay and reproduction conditions . Results are compared with those obtained using the checkerboard method and mortality-time curves . Trials were carried out with a strain of Candida albicans (ATCC 90028) . The E-test strips were combined in direct proportion, MIC-MIC, and in inverse proportion . The results showed that the method can be reproduced, is easy to carry out and may be suitable for the study of the in vitro interaction of antimycotics on yeast . The directly and inversely proportionate strip combination appeared to be the most suitable . At the prediffusion stage, the most highly water-soluble antimycotic should be used . The recommended time for prediffusion is one hour for water-soluble antimycotics, and two hours for non-water-soluble ones . The E-test showed good correlation with mortality-time curves . Nonetheless, in vivo correlation studies are required to determine the usefulness of the results in vitro and the most suitable method of measurement

J Antimicrob Chemother, 2004 Aug, 54(2), 376 - 85 Epub 2004 Jun 16.
Genome-wide expression profiling reveals genes associated with amphotericin B and fluconazole resistance in experimentally induced antifungal resistant isolates of Candida albicans; Barker KS et al.; OBJECTIVES: The aim of this study was to identify changes in the gene expression profile of Candida albicans associated with the acquisition of experimentally induced resistance to amphotericin B and fluconazole . METHODS: C . albicans SC5314 was passed in increasing concentrations of amphotericin B to generate isolate SC5314-AR . Susceptibility testing by Etest revealed SC5314-AR to be highly resistant to both amphotericin B and fluconazole . The gene expression profile of SC5314-AR was compared with that of SC5314 using DNA microarray analysis . Sterol composition was determined for both strains . RESULTS: Upon examination of MICs of antifungal compounds, it was found that SC5314-AR was resistant to both amphotericin B and fluconazole . By microarray analysis a total of 134 genes were found to be differentially expressed, that is up-regulated or down-regulated by at least 50%, in SC5314-AR . In addition to the cell stress genes DDR48 and RTA2, the ergosterol biosynthesis genes ERG5, ERG6 and ERG25 were up-regulated . Several histone genes, protein synthesis genes and energy generation genes were down-regulated . Sterol analysis revealed the prevalence of sterol intermediates eburicol and lanosterol in SC5314-AR, whereas ergosterol was the predominant sterol in SC5314 . CONCLUSION: Along with changes in expression of these ergosterol biosynthesis genes was the accumulation of sterol intermediates in the resistant strain, which would account for the decreased affinity of amphotericin B for membrane sterols and a decreased requirement for lanosterol demethylase activity in membrane sterol production . Furthermore, other genes are implicated as having a potential role in the polyene and azole antifungal resistant phenotype.

Yeast, 2004 Jun, 21(8), 685 - 702
Role of the 14-3-3 protein in carbon metabolism of the pathogenic yeast Candida albicans; Wang YK et al.; We previously demonstrated that the pathogenic yeast Candida albicans effectively adapts to utilize L-sorbose (Sou+) by a novel mechanism based on the loss of one copy of chromosome 5, probably due to the reduction of copy number of a negative regulator located on this chromosome . We report here another negative regulator of L-sorbose utilization, an orthologue of the Saccharomyces cerevisiae BMH1 gene, which encodes the evolutionarily conserved protein 14-3-3 . This essential gene is located on chromosome 1, does not have paralogues, and is supposedly a component of the regulatory network . Experiments involving disruption of one allele of BMH1 and overexpression of BMH1 revealed that BMH1 represses the transcription of SOU1, which is responsible for the utilization of L-sorbose . Although the exact mechanism of the interaction between BMH1 and SOU1 is not known, it is clear that the control is based on the ratio of gene copy number, and that BMH1 does not control the loss of chromosome 5, the major mechanism producing Sou+ mutants . We propose that function of BMH1 as a negative regulator of SOU1 contributes to a general cellular homeostasis . This is a first report on the role of the C . albicans essential gene BMH1 as a negative regulator of the utilization of secondary carbon source in yeast, which further substantiates the involvement of 14-3-3 proteins in diverse functions .

Yeast, 2004 Jun, 21(8), 619 - 34
Chromosome translocation induced by the insertion of the URA blaster into the major repeat sequence (MRS) in Candida albicans; Iwaguchi S et al.; Electrophoretic karyotype studies have shown that clinical isolates of Candida albicans have extensive chromosome length polymorphisms . Chromosome translocation is one of the causes of karyotypic variation . Chromosome translocation events have been shown to occur very frequently at or near the major repeat sequence (MRS) on chromosomes . The MRS consists of the repeated sequences RB2, RPS and HOK, and the repeated sequences are considered to be the template for recombination . To investigate which element of the MRS is important for chromosome translocation, we constructed three cassettes, each containing a URA blaster and sequences homologous to one of the repeats, for insertion into the MRS region on the chromosomes . The ura3 strain STN22u2, which shows a stable, standard karyotype, was transformed with each construct . Insertion events with each cassette occurred at almost all chromosomes . Insertion into the RB2 repeat, but not into the RPS repeat, was accompanied by chromosome translocation in some transformants: chromosome translocations between chromosomes R and 7 and chromosomes 1 and 7 were found, as well as deletions of 7A and 7C from chromosome 7 . We conclude that the insertion at the RB2 region may initiate chromosome translocation in C . albicans .

FEMS Immunol Med Microbiol, 2004 Jul 1, 41(3), 249 - 58
Co-administration of immunomodulator tuftsin and liposomised nystatin can combat less susceptible Candida albicans infection in temporarily neutropenic mice; Khan MA et al.; In order to develop a prospective chemotherapeutic agent against opportunistic infections, it is important to know that host factors such as degree of immunological debility as well as recovery of immune functions to normality may contribute significantly to a successful elimination of the pathogens . We demonstrated previously that concomitant delivery of antimicrobial agents and immunomodulators to the pathogen harbouring-host contributes to the complete elimination of the deep-seated fungal infections (aspergillosis and candidiasis) in animals with normal immune status . Considering that neutropenic hosts are the main targets of such infections, it can be argued about the potential of the immunomodulator-based therapy in subjects with non-functional immune system . To resolve the hypothesis, we studied the role of immunomodulator tuftsin against experimental murine candidiasis in temporarily neutropenic Balb/c mice . The neutropenic mice were challenged with an isolate of Candida albicans that was showing less susceptibility to both free and liposomised-amphotericin B . The co-administration of tuftsin increased the efficiency of liposomised-polyene antibiotics (nystatin and amphotericin B) against experimental murine candidiasis in immunocompromised Balb/c mice . Pretreatment with liposomised tuftsin prior to C . albicans infection clearly enhanced protection against candidiasis, suggesting a prophylactic role of tuftsin in normal and temporarily neutropenic animals.

FEMS Immunol Med Microbiol, 2004 Jul 1, 41(3), 227 - 35
Candida albicans and Saccharomyces cerevisiae induce interleukin-8 production from intestinal epithelial-like Caco-2 cells in the presence of butyric acid; Saegusa S et al.; Intestinal epithelial cells (IEC) are important in initiation and regulation of immune responses against numerous foreign substances including food, microorganisms and their metabolites in the intestine . Since the responses of IEC against yeasts have not yet been well understood, we investigated the effects of Candida albicans, Saccharomyces cerevisiae, and their cell wall components on interleukin-8 (IL-8) secretion by the IEC-like Caco-2 cells . Live cells of both yeast species stimulated Caco-2 cells to produce IL-8 only in the presence of butyric acid, which is a metabolite produced by intestinal bacteria . S . cerevisiae zymosan and glucan also enhanced IL-8 secretion . Treatment of Caco-2 cells with butyric acid increased the expression of mRNAs coding for Toll-like receptor 1 (TLR1), TLR6 and dectin-1, which recognize zymosan . C . albicans induced more IL-8 secretion and also decreased transepithelial electrical resistance more rapidly than S . cerevisiae . These results suggest that both yeasts in the intestine stimulate the host's mucosal immune systems by interacting with IEC.

FEMS Immunol Med Microbiol, 2004 Jul 1, 41(3), 187 - 96
Antibody response to Candida albicans cell wall antigens; Lopez-Ribot JL et al.; The cell wall of Candida albicans is not only the structure where many essential biological functions reside but is also a significant source of candidal antigens . The major cell wall components that elicit a response from the host immune system are proteins and glycoproteins, the latter being predominantly mannoproteins . Both carbohydrate and protein moieties are able to trigger immune responses . Proteins and glycoproteins exposed at the most external layers of the wall structure are involved in several types of interactions of fungal cells with the exocellular environment . Thus, coating of fungal cells with host antibodies has the potential to profoundly influence the host-parasite interaction by affecting antibody-mediated functions such as opsonin-enhanced phagocytosis and blocking the binding activity of fungal adhesins to host ligands . In this review we examine various members of the protein and glycoprotein fraction of the C . albicans cell wall that elicit an antibody response in vivo . Some of the studies demonstrate that certain cell wall antigens and anti-cell wall antibodies may be the basis for developing specific and sensitive serologic tests for the diagnosis of candidiasis, particularly the disseminated form . In addition, recent studies have focused on the potential of antibodies against the cell wall protein determinants in protecting the host against infection . Hence, a better understanding of the humoral response triggered by the cell wall antigens of C . albicans may provide the basis for the development of (i) effective procedures for the serodiagnosis of disseminated candidiasis, and (ii) novel prophylactic (vaccination) and therapeutic strategies to control this type of infections.

Oral Dis, 2004 Jul, 10(4), 221 - 8
Fungicidal effect of three new synthetic cationic peptides against Candida albicans; Nikawa H et al.; OBJECTIVE: Peptide antibiotics are considered a new class of antifungal agents . Of these, an alpha-helical, cationic peptide termed Dhvar 4, a relative of salivary histatin has been shown to be an antifungal of relatively high potency . Similarly, lactoferricin B (LFB) and a derivative thereof, LFB(17-30), disrupts the fungal cell membrane and acts against Candida albicans . As Dhvar 4 and LFB(17-30), exhibit almost identical amino acid sequences at their C-terminal, we hypothesized that laboratory synthesis of peptides with an alpha-helical structure and having similar amphipathic properties could lead to products with candidacidal activity . Hence, three such peptides - JH8194, JH8195 and JH 8944, were synthesized and their antifungal properties compared with recognized antifungals LFB, LFB(17-30), human lactoferricin (LFH), Histatin-5 and Dhvar 4, against two isolates of C . albicans . MATERIALS AND METHODS: The antifungal agents were synthesized and their secondary structures evaluated according to a previously described protocol of Situ and Bobek (2000)Antimicrob Agents Chemother44: 1485-1493 . The C . albicans strains were oral isolates from a human immunodeficiency virus-infected (isolate A2) and a healthy (A6) individual . A standard concentration of yeasts was exposed to a range of dilutions of the agents for a specific duration and the cell death (viability) in terms of the resultant colony forming units ml(-1) was quantified . RESULTS: Dhvar 4, showed the most alpha-helical propensity, and was the least fungicidal while LFB and LFB(17-30) showed the highest antifungal potential, and demonstrated total kill of A6, and A2 at 5 and 10 microM concentrations, respectively whilst LFH killed both isolates at a l0 microM concentration . Of the three new synthetic peptides, JH 8194 was the most potent (total kill of A6/A2 strains at 1.25/2.5 microM), followed by JH 8195 (total kill of A6/A2 strains at 5/10 microM while JH 8944 was the least potent as a 25 microM concentration was required to kill either strain of Candida . On further analyses of the relationship between pI value of the peptides and their anticandicidal activity, a significant positive correlation was noted . In order to rule out a cytotoxic effect of the new synthetic peptides we compared the fungicidal and hemolytic activities under similar incubation conditions using freshly isolated erythrocytes and all three peptides exhibited no detectable hemolysis upto an concentration of 100 microM in contrast to the polyene antifungal amphotericin B that elicited significant initiation of hemolysis at a concentration of 5.0 microM . CONCLUSION: Our data suggest that laboratory synthesis of agents with an alpha-helical structure and having amphipathic properties similar to known, natural antifungal agents may be a promising avenue to generate products with improved antifungal activity.

Acta Cytol, 2004 May-Jun, 48(3), 407 - 10
Cytologic diagnosis of suppurative cholecystitis due to Candida albicans and actinomyces . A report of 2 cases; Santos LD et al.; BACKGROUND: Cholecystitis is a common inflammatory disease of the gallbladder . Actinomycosis and candidiasis of the gallbladder are uncommon causes of acute cholecystitis . There has been no previous report on the cytologic diagnosis of actinomycosis and candidiasis from aspirated gallbladder bile intraoperatively . CASES: Purulent bile was intraoperatively aspirated from the gallbladder of 71-year-old Indian and a 30-year-old Australian woman . The specimens were sent for cytologic examination . The first case revealed sulphur granules characteristic of Actinomyces spp . The second case showed budding spores and pseudohyphae of Candida spp . Pure colonies of Candida albicans grew from the bile culture . CONCLUSION: Actinomycosis and candidiasis rarely cause acute suppurative cholecystitis . Initial diagnosis can be made by cytologic examination of the aspirated purulent bile intraoperatively.

Med J Malaysia, 2003 Dec, 58(5), 788 - 92
Recurrent vaginal candidiasis; Loh KY et al.; Recurrent vaginal candidiasis is one of the most common reasons for patients visiting their primary care doctors . Majority of the cases are caused by Candida albicans . Controlling of risk factors such as diabetes mellitus, used of broad spectrum antibiotics, contraceptive pills and steroid therapy helps in managing recurrent vaginal candidiasis . Initial 14-day course of oral azoles and followed by 6 months maintenance are effective in treating majority of the cases . Failure to treat recurrent vaginal candidiasis can lead to various bio-psycho-social complications.

Rev Port Cir Cardiotorac Vasc, 2004 Jan-Mar, 11(1), 47 - 50
{Mycotic rupture of the reno-external iliac artery anastomosi following kidney transplantation}; Maio R et al.; Two clinical cases of an unusual complication of the renal transplantation are reported, consisting in the infection of the arterial wall by fungi of the species Candida Albicans and Aspergillus, which caused acute ruptures of the external iliac artery and hypovolemic shock . The main features of the pathogenesis of the infection, its diagnosis, management and prevention are subjected to analysis and discussion.

J Antimicrob Chemother, 2004 Jul, 54(1), 38 - 45 Epub 2004 Jun 09.
Substitution of threonine-1351 in the multidrug transporter Cdr1p of Candida albicans results in hypersusceptibility to antifungal agents and threonine-1351 is essential for synergic effects of calcineurin inhibitor FK520; Shukla S et al.; OBJECTIVES: Functional characterization of a mutant Candida albicans drug resistance protein (Cdr1p) by overexpression in Saccharomyces cerevisiae . METHODS: We overexpressed green fluorescent protein-tagged Cdr1p in S . cerevisiae AD1-8u- host and introduced a point mutation to substitute T1351 with F in Cdr1p . The cells expressing T1351F mutant Cdr1p were analysed for their functional activity using minimum inhibitory concentration, spot assay, and fluconazole efflux . The binding activity of photoaffinity analogues 8-azidoATP, iodoarylazidoprazosin and azidopine to the mutant T1351F Cdr1p was also characterized . RESULTS: The T1351F mutant Cdr1p-expressing cells were susceptible to anisomycin, cycloheximide, fluconazole, miconazole and nystatin . The mutant protein was expressed to the same level as that of native Cdr1p in S . cerevisiae cells and was properly localized to the cell surface . There was also no difference between the mutant variant and the native protein's ability to bind a photoaffinity analogue of ATP, 8-azidoATP, or the radiolabelled photoaffinity agents iodoarylazidoprazosin and azidopine . However, the substitution of T1351 resulted in considerable reduction in its ability to export fluorescent substrate rhodamine 6G . The synergy between calcineurin inhibitors FK520 and azoles was abrogated in cells expressing the T1351F mutant variant of Cdr1p . CONCLUSIONS: The results from this study suggest that the T1351 in the predicted transmembrane domain (TMD) 11 of Cdr1p is not only important for drug-substrate transport but also has a role in governing synergy of FK520.

Eukaryot Cell, 2004 Jun, 3(3), 776 - 84
RBR1, a novel pH-regulated cell wall gene of Candida albicans, is repressed by RIM101 and activated by NRG1; Lotz H et al.; The transcription factor Rim101p of Candida albicans has been shown to play a major role in pH-dependent gene regulation . Rim101p is involved in cell wall biosynthesis, since it regulates PHR1 and PHR2, two almost functionally redundant cell wall glycosidases important for adaptation to either neutral or acidic habitats within the human host . To identify additional cell wall components regulated by Rim101p, we performed transcriptional profiling with a cell wall-specific DNA microarray . We showed that Rim101p contributes to the activation of known hypha-specific genes such as HWP1 and RBT1 but is also required for repression of the previously uncharacterized potential cell wall genes RBR1, RBR2, and RBR3 . Further characterization of RBR1 revealed that it encodes a small glycosylphosphatidyl inositol protein that is expressed under acidic conditions predominantly at low temperature . Deletion of the gene resulted in a filamentation defect at low pH . Most interestingly, NRG1, a transcriptional repressor of hyphal growth in C . albicans, was required for RBR1 expression . The apparently activating effect of NRG1 observed in this study has not been described before . In addition, we showed that expression of NRG1 is not only temperature but also pH dependent .

Eukaryot Cell, 2004 Jun, 3(3), 764 - 75
Hemoglobin regulates expression of an activator of mating-type locus alpha genes in Candida albicans; Pendrak ML et al.; Phenotypic switching from the white to the opaque phase is a necessary step for mating in the pathogenic fungus Candida albicans . Suppressing switching during vascular dissemination of the organism may be advantageous, because opaque cells are more susceptible to host defenses . A repressor of white-opaque switching, HBR1 (hemoglobin response gene 1), was identified based on its specific induction following growth in the presence of exogenous hemoglobin . Deletion of a single HBR1 allele allowed opaque phase switching and mating competence, accompanied by a lack of detectable MTL alpha1 and alpha2 gene expression and enhanced MTLa1 gene expression . Conversely, overexpression of Hbr1p or exposure to hemoglobin increased MTLalpha gene expression . The a1/alpha2 repressed target gene CAG1 was derepressed in the same mutant in a hemoglobin-sensitive manner . Regulation of CAG1 by hemoglobin required an intact MTLa1 gene . Several additional Mtlp targets were perturbed in HBR1 mutants in a manner consistent with commitment to an a mating phenotype, including YEL007w, MFalpha, HST6, and RAM2 . Therefore, Hbr1 is part of a host factor-regulated signaling pathway that controls white-opaque switching and mating in the absence of allelic deletion at the MTL locus .

Eukaryot Cell, 2004 Jun, 3(3), 741 - 51
Candida albicans Rim13p, a protease required for Rim101p processing at acidic and alkaline pHs; Li M et al.; Candida albicans is an important commensal of mucosal surfaces that is also an opportunistic pathogen . This organism colonizes a wide range of host sites that differ in pH; thus, it must respond appropriately to this environmental stress to survive . The ability to respond to neutral-to-alkaline pHs is governed in part by the RIM101 signal transduction pathway . Here we describe the analysis of C . albicans Rim13p, a homolog of the Rim13p/PalB calpain-like protease member of the RIM101/pacC pathway from Saccharomyces cerevisiae and Aspergillus nidulans, respectively . RIM13, like other members of the RIM101 pathway, is required for alkaline pH-induced filamentation and growth under extreme alkaline conditions . Further, our studies suggest that the RIM101 pathway promotes pH-independent responses, including resistance to high concentrations of lithium and to the drug hygromycin B . RIM13 encodes a calpain-like protease, and we found that Rim101p undergoes a Rim13p-dependent C-terminal proteolytic processing event at neutral-to-alkaline pHs, similar to that reported for S . cerevisiae Rim101p and A . nidulans PacC . However, we present evidence that suggests that C . albicans Rim101p undergoes a novel processing event at acidic pHs that has not been reported in either S . cerevisiae or A . nidulans . Thus, our results provide a framework to understand how the C . albicans Rim101p processing pathway promotes alkaline pH-independent processes .

Eukaryot Cell, 2004 Jun, 3(3), 724 - 34
Cdc42p GTPase regulates the budded-to-hyphal-form transition and expression of hypha-specific transcripts in Candida albicans; VandenBerg AL et al.; The yeast Candida albicans is a major opportunistic pathogen of immunocompromised individuals . It can grow in several distinct morphological states, including budded and hyphal forms, and the ability to make the dynamic transition between these forms is strongly correlated with virulence . Recent studies implicating the Cdc42p GTPase in hypha formation relied on cdc42 mutations that affected the mitotic functions of the protein, thereby precluding any substantive conclusions about the specific role of Cdc42p in the budded-to-hypha-form transition and virulence . Therefore, we took advantage of several Saccharomyces cerevisiae cdc42 mutants that separated Cdc42p's mitotic functions away from its role in filamentous growth . The homologous cdc42-S26I, cdc42-E100G, and cdc42-S158T mutations in C . albicans Cdc42p caused a dramatic defect in the budded-to-hypha-form transition in response to various hypha-inducing signals without affecting normal budded growth, strongly supporting the conclusion that Cdc42p has an integral function in orchestrating the morphological transition in C . albicans . In addition, the cdc42-S26I and cdc42-E100G mutants demonstrated a reduced ability to damage endothelial cells, a process that is strongly correlated to virulence . The three mutants also had reduced expression of several hypha-specific genes, including those under the regulation of the Efg1p transcription factor . These data indicate that Cdc42p-dependent signaling pathways regulate the budded-to-hypha-form transition and the expression of hypha-specific genes .

Eukaryot Cell, 2004 Jun, 3(3), 715 - 23
Inducible defense mechanism against nitric oxide in Candida albicans; Ullmann BD et al.; The yeast Candida albicans is an opportunistic pathogen that threatens patients with compromised immune systems . Immune cell defenses against C . albicans are complex but typically involve the production of reactive oxygen species and nitrogen radicals such as nitric oxide (NO) that damage the yeast or inhibit its growth . Whether Candida defends itself against NO and the molecules responsible for this defense have yet to be determined . The defense against NO in various bacteria and the yeast Saccharomyces cerevisiae involves an NO-scavenging flavohemoglobin . The C . albicans genome contains three genes encoding flavohemoglobin-related proteins, CaYHB1, CaYHB4, and CaYHB5 . To assess their roles in NO metabolism, we constructed strains lacking each of these genes and demonstrated that just one, CaYHB1, is responsible for NO consumption and detoxification . In C . albicans, NO metabolic activity and CaYHB1 mRNA levels are rapidly induced by NO and NO-generating agents . Loss of CaYHB1 increases the sensitivity of C . albicans to NO-mediated growth inhibition . In mice, infections with Candida strains lacking CaYHB1 still resulted in lethality, but virulence was decreased compared to that in wild-type strains . Thus, C . albicans possesses a rapid, specific, and highly inducible NO defense mechanism involving one of three putative flavohemoglobin genes .

Eukaryot Cell, 2004 Jun, 3(3), 705 - 14
Genome-wide single-nucleotide polymorphism map for Candida albicans; Forche A et al.; Single-nucleotide polymorphisms (SNPs) are essential tools for studying a variety of organismal properties and processes, such as recombination, chromosomal dynamics, and genome rearrangement . This paper describes the development of a genome-wide SNP map for Candida albicans to study mitotic recombination and chromosome loss . C . albicans is a diploid yeast which propagates primarily by clonal mitotic division . It is the leading fungal pathogen that causes infections in humans, ranging from mild superficial lesions in healthy individuals to severe, life-threatening diseases in patients with suppressed immune systems . The SNP map contains 150 marker sequences comprising 561 SNPs and 9 insertions-deletions . Of the 561 SNPs, 437 were transition events while 126 were transversion events, yielding a transition-to-transversion ratio of 3:1, as expected for a neutral accumulation of mutations . The average SNP frequency for our data set was 1 SNP per 83 bp . The map has one marker placed every 111 kb, on average, across the 16-Mb genome . For marker sequences located partially or completely within coding regions, most contained one or more nonsynonymous substitutions . Using the SNP markers, we identified a loss of heterozygosity over large chromosomal fragments in strains of C . albicans that are frequently used for gene manipulation experiments . The SNP map will be useful for understanding the role of heterozygosity and genome rearrangement in the response of C . albicans to host environments .

Eukaryot Cell, 2004 Jun, 3(3), 675 - 84
Lipid raft polarization contributes to hyphal growth in Candida albicans; Martin SW et al.; The polarization of sterol- and sphingolipid-enriched domains (lipid rafts) has been linked to morphogenesis and cell movement in diverse cell types . In the yeast Saccharomyces cerevisiae, a dramatic polarization of sterol-rich domains to the shmoo tip was observed in pheromone-induced cells (M . Bagnat and K . Simons, Proc . Natl . Acad . Sci . USA 99:14183-14188, 2002) . We therefore examined whether plasma membrane lipid polarization contributes to the ability of the fungal pathogen Candida albicans to grow in a highly polarized manner to form hyphae . Interestingly, staining with filipin revealed that membrane sterols were highly polarized to the leading edge of growth during all stages of hyphal growth . Budding and pseudohyphal cells did not display polarized staining . Filipin staining was also enriched at septation sites in hyphae, where colocalization with septin proteins was observed, suggesting a role for the septins in forming a boundary domain . Actin appeared to play a role in sterol polarization and hyphal morphogenesis in that both were disrupted by low concentrations of latrunculin A that did not prevent budding . Furthermore, blocking either sphingolipid biosynthesis with myriocin or sterol biosynthesis with ketoconazole resulted in a loss of ergosterol polarization and caused abnormal hyphal morphogenesis, suggesting that lipid rafts are involved . Since hyphal growth is required for the full virulence of C . albicans, these results suggest that membrane polarization may contribute to the pathogenesis of this organism .

Mycoses, 2004 Jun, 47(5-6), 193 - 9
Candida albicans-induced inflammatory response in human keratinocytes; Wollina U et al.; Candida albicans strains 3153a, ATCC 48867, CBS 2730, DSM 70014, and Vir 13 were cultivated and sterile C . albicans filtrates were produced . The interaction of soluble Candida factors of these infiltrates with human HaCaT keratinocytes was assayed in vitro . The following parameters were analyzed: cell proliferation, protein synthesis, nuclear matrix protein (NMP) 41 release, cytokine release (IL-1beta, soluble IL-2 receptor, IL-6, and IL-8), and reactive oxygen species (ROS) . Cell counts at 1, 12, and 24 h were significantly lower for C . albicans strains CBS 2730 and VIR 13 (P < 0.05) . There was no significant change for the remaining strains . Neither the protein synthesis nor the NMP-41 release was significantly affected . IL-6 and IL-8 were stimulated by C . albicans filtrates to different amounts with higher levels in strains of low virulence . There was no effect on the other cytokines . The production of ROS by HaCaT keratinocytes was suppressed . The induction of an inflammatory keratinocyte response by soluble C . albicans factors may play a role among the host-yeast interactions.

Mycoses, 2004 Jun, 47(5-6), 184 - 92
Agar sublimation test for the in vitro determination of the antifungal activity of morpholine derivatives; Polak A et al.; We studied the in vitro antifungal activities of a wide range of antimycotic agents, including amorolfine, terbinafine, naftifine, five morpholine derivatives, ciclopiroxolamine, bifonazole, clotrimazole, ketoconazole, itraconazole, fluconazole, voriconazole, flucytosine, amphotericin B, nystatin, and caspofungin, against Candida albicans and Trichophyton rubrum by conventional agar diffusion tests and by a novel sublimation method . For the sublimation method, 6 mm filter paper disks were soaked with defined amounts of antimycotic drugs, air dried, placed in the center of the lids of 9 cm Petri dishes, and incubated upside down with inoculated agar plates 10 mm above the disks . The conventional disk diffusion tests produced inhibition zones as previously described . The disk sublimation tests produced large inhibition zones with amorolfine, five amorolfine derivatives, and terbinafine, but with none of the other antifungal agents . Possible therapeutic advantages of agents, which are able to overcome air cavities in mycotic lesions, e.g . in onychomycosis, are discussed.

Biol Pharm Bull, 2004 Jun, 27(6), 890 - 2
Hyphae formation of Candida albicans is regulated by polyamines; Ueno Y et al.; Candida albicans generally grows in hyphae form in RPMI1640 medium . However, addition of 1,4-diamino-2-butanone (DAB), a competitive inhibitor of ornithine decarboxylase, decreased the amount of polyamines in C . albicans, and induced the proliferation of the C . albicans yeast form . The expression of CYR1 mRNA was significantly inhibited by the addition of DAB compared with that of the control . The amount of intracellular cAMP was also decreased by the addition of DAB . A specific adenylate cyclase inhibitor, cis-N-{2-phenylcyclopentyl}-azacyclotridec-1-en-2-amine (MDL-12,330A) promoted the growth of the yeast form . These results indicated that polyamines exist upstream of the adenylate cyclase-cAMP signal pathway and regulate the transformation of C . albicans.

Microbiology, 2004 Jun, 150(Pt 6), 1991 - 2003
Dosage-dependent functions of fatty acid desaturase Ole1p in growth and morphogenesis of Candida albicans; Krishnamurthy S et al.; Conditions in the infected human host trigger virulence attributes of the fungal pathogen Candida albicans . Specific inducers and elevated temperatures lead to hyphal development or regulate chlamydospore development . To explore if these processes are affected by membrane lipids, an investigation of the functions of the Ole1 fatty acid desaturase (stearoyl-CoA desaturase) in C . albicans, which synthesizes oleic acid, was undertaken . A conditional strain expressing OLE1 from the regulatable MET3 promoter was unable to grow in repressing conditions, indicating that OLE1 is an essential gene . In contrast, a mutant lacking both alleles of OLE2, encoding a Ole1p homologue, was viable and had no apparent phenotypes . Partial repression of MET3p-OLE1 slightly lowered oleic acid levels and decreased membrane fluidity; these conditions permitted growth in the yeast form, but prevented hyphal development in aerobic conditions and blocked the formation of chlamydospores . In contrast, in hypoxic conditions, which trigger an alternative morphogenetic pathway, hyphal morphogenesis was unaffected . Because aerobic morphogenetic signalling and oleic acid biosynthesis require oxygen, it is proposed that oleic acid may function as a sensor activating specific morphogenetic pathways in normoxic conditions.

Microbiology, 2004 Jun, 150(Pt 6), 1911 - 24
Mutant alleles of the essential 14-3-3 gene in Candida albicans distinguish between growth and filamentation; Palmer GE et al.; The opportunistic fungal pathogen Candida albicans has the ability to exploit diverse host environments and can either reside commensally or cause disease . In order to adapt to its new environment it must respond to new physical conditions, nutrient sources, and the host immune response . This requires the co-regulation of multiple signalling networks . The 14-3-3 family of proteins is highly conserved in all eukaryotic species . These proteins regulate signalling pathways involved in cell survival, the cell cycle, and differentiation, and effect their functions via interactions with phosphorylated serines/threonines . In C . albicans there is only one 14-3-3 protein, Bmh1p, and it is required for vegetative growth and optimal filamentation . In order to dissect separate functions of Bmh1p in C . albicans, site-directed nucleotide substitutions were made in the C . albicans BMH1 gene based on studies in other species . Putative temperature-sensitive, ligand-binding and dimerization mutants were constructed . In addition two mutant strains identified through random mutagenesis were analysed . All five mutant strains demonstrated varying defects in growth and filamentation . This paper begins to segregate functions of Bmh1p that are required for optimal growth and the different filamentation pathways . These mutant strains will allow the identification of 14-3-3 target interactions and correlate the individual functions of Bmh1p to cellular processes involved in pathogenesis.

J Med Microbiol, 2004 Jul, 53(Pt 7), 633 - 7
Isolation and molecular identification of Candida dubliniensis from non-human immunodeficiency virus-infected patients in Kuwait; Ahmad S et al.; Candida dubliniensis is an emerging pathogen capable of causing oropharyngeal, vaginal and bloodstream infections . Although C . dubliniensis is similar to Candida albicans in several phenotypic characteristics, it differs from it with respect to epidemiology, certain virulence factors and the ability to develop resistance to fluconazole rapidly . In this study, the first seven isolations of C . dubliniensis from Kuwait are described, all originating from non-human immunodeficiency virus (HIV)-infected patients . The isolates were initially identified by the Vitek 2 yeast identification system, positive germ tube test, production of rough colonies and chlamydospores on Staib agar and by their inability to assimilate xylose, trehalose or methyl alpha-D-glucoside . The species identity of the isolates was subsequently confirmed by specific amplification of rDNA targeting the internally transcribed spacer 2 (ITS2), restriction endonuclease digestion of the amplified DNA and direct DNA sequencing of the ITS2 . Using the E-test method, the MICs of C . dubliniensis test isolates were in the range 0.125-0.75 microg ml(-1) for fluconazole, 0.002-0.75 microg ml(-1) for itraconazole, 0.006-0.125 microg ml(-1) for ketoconazole, 0.002-0.5 microg ml(-1) for amphotericin B and 0.002-0.016 microg ml(-1) for voriconazole . Two of the isolates were resistant to 5-flucytosine (>32 microg ml(-1)), but none against fluconazole . The study reinforces the current view that C . dubliniensis has a much wider geographical and epidemiological distribution.

J Clin Microbiol, 2004 Jun, 42(6), 2480 - 8
Multilocus sequence typing is a reliable alternative method to DNA fingerprinting for discriminating among strains of Candida albicans; Robles JC et al.; Multilocus sequence typing (MLST) has emerged as a powerful new DNA-typing tool for the evaluation of intraspecies genetic relatedness . This method relies on DNA sequence analysis of nucleotide polymorphisms in housekeeping genes and has shown a high degree of intraspecies discriminatory power for bacterial and fungal pathogens . However, the results of the MLST scheme for Candida albicans have heretofore never been formally compared to those of other established typing techniques . To assess the value of MLST relative to those of other DNA fingerprinting tools for discriminating among strains of C . albicans, we applied it to a previously well-characterized set of 29 C . albicans isolates evaluated by the random amplified polymorphic DNA (RAPD), multilocus enzyme electrophoresis (MLEE), and Ca3 Southern hybridization probe techniques . MLST identified three clusters of genetically related isolates, with 82.3% direct concordance with MLEE, 82.7% with RAPD analysis, and 86.2% with the Ca3 Southern hybridization technique . When MLST was applied to a subset of 22 isolates of unrelated origins, it identified 21 independent diploid sequence types (DSTs), resulting in a discriminatory power of 99.6% . These DSTs were 96.9, 99.6, and 99.6% concordant with the genotypes identified by RAPD analysis, MLEE, and Ca3 Southern hybridization, respectively . These results demonstrate that MLST is a highly effective technique that performs at least comparably to other established DNA fingerprinting techniques.

Biochem Biophys Res Commun, 2004 Jul 2, 319(3), 911 - 9
Molecular cloning and biochemical characterization of Candida albicans acyl-CoA:sterol acyltransferase, a potential target of antifungal agents; Kim KY et al.; To determine whether Candida albicans acyl CoA:sterol acyltransferase (ASAT) can be a potential target enzyme for the protoberberine derivative (HWY-289), we have isolated a gene encoding Ca-ASAT and examined inhibitory effects of HWY-289 on the overexpressed Ca-ASAT . HWY-289 specifically inhibits Ca-ASAT in a non-competitive manner in vitro (IC(50) {9.2microM}, K(i) {5.15microM}) . The cloned CaARE2 gene (1830 nucleotides {nt}) encodes active Ca-ASAT protein that exhibits a calculated molecular mass of 71.3kDa . The amino acid sequence of CaAre2p is 33.4% and 35.1% identical to those of Saccharomyces cerevisiae ScAre1p and ScAre2p homologues, respectively . Recombinant and endogenous Ca-ASAT displayed identical patterns of inhibition upon exposure to HWY-289 and a preference for cholesterol and oleoyl-CoA as substrates . Northern blot analysis showed that CaARE2 was activated by HWY-289, but not by CI-976 (a human acyl-coenzyme A:cholesterol acyltransferase inhibitor), in a dose-dependent manner (up to 5mg/L), suggesting different selectivities of action between HWY-289 and CI-976 on Ca-ASAT activity.

FEMS Microbiol Lett, 2004 Jun 15, 235(2), 297 - 303
Identification of an exoribonuclease homolog, CaKEM1/CaXRN1, in Candida albicans and its characterization in filamentous growth; An HS et al.; KEM1/XRN1 and RAT1 are two known exoribonuclease genes in Saccharomyces cereivsiae and encode a cytoplasmic and nuclear exoribonuclease, respectively . CaKEM1/CaXRN1 and CaRAT1, the Candida albicans homologs of 5'-->3' exoribonuclease genes, were identified by protein sequence comparisons and by functional complementation of the S . cerevisiae kem1/xrn1 null mutation . The deduced amino acid sequences of CaKEM1 and CaRAT1 show 51% and 55% identities to those of the S . cerevisiae KEM1 and RAT1, respectively . The exonuclease motifs were found to be highly conserved in CaKem1p and CaRat1p . We disrupted two chromosomal copies of CaKEM1 in a diploid C . albicans strain and demonstrate that C . albicans kem1/kem1 mutants are defective in filamentous growth on filamentous-inducing media . These results imply that CaKEM1 is involved in filamentous growth of C . albicans.

Biochim Biophys Acta, 2004 Jun 11, 1672(3), 214 - 23
Overexpression, purification and characterization of the acidic ribosomal P-proteins from Candida albicans; Abramczyk D et al.; In all eukaryotic cells, acidic ribosomal P-proteins form a lateral protuberance on the 60S ribosomal subunit-the so-called stalk-structure that plays an important role during protein synthesis . In this work, we report for the first time a full-length cloning of four genes encoding the P-proteins from Candida albicans, their expression in Escherichia coli, purification and characterization of the recombinant proteins . Considerable amino acid sequence similarity was found between the cloned proteins and other known fungal ribosomal P-proteins . On the basis of their phylogenetic relationship and amino acid similarity to their yeast counterparts, the C . albicans P-proteins were named P1A, P1B, P2A and P2B . Using three different approaches, namely: chemical cross-linking method, gel filtration and two-hybrid system, we analyzed mutual interactions among the C . albicans P-proteins . The obtained data showed all the four P-proteins able to form homo-oligomeric complexes . However, the ones found between P1B-P2A and P1A-P2B were dominant forms among the C . albicans P-proteins . Moreover, the strength of interactions between particular proteins was different in these two complexes; the strongest interactions were observed between P1B and P2A proteins, and a significantly weaker one between P1A and P2B proteins.

J Microbiol Immunol Infect, 2004 Apr, 37(2), 115 - 20
Fungal peritonitis in peritoneal dialysis patients: effect of fluconazole treatment and use of the twin-bag disconnect system; Chen CM et al.; Fungal peritonitis is an uncommon but potentially life-threatening complication for patients undergoing continuous ambulatory peritoneal dialysis . This retrospective study evaluated the efficacy of fluconazole in fungal peritonitis treatment and the incidence of fungal peritonitis in different peritoneal dialysis disconnect systems . Fungal peritonitis was caused by Candida species in 67% of episodes . The most common pathogen in this series was Candida parapsilosis (29%), followed by Candida albicans (14%) . One patient (5%) died within 1 month after admission for treatment of fungal peritonitis . Only 1 patient (5%) in this series could resume peritoneal dialysis . Treatment with fluconazole alone has an effect comparable to intraperitoneal (IP) amphotericin B alone or IP amphotericin B combined with oral or intravenous fluconazole . The incidence of fungal peritonitis in patients who used the spike, Y-set, and UV antiseptic systems was 5.69, 6.20, and 2.93 times, respectively, as frequent as that of fungal peritonitis in patients who used the twin-bag disconnect system.

Curr Drug Targets Infect Disord, 2004 Jun, 4(2), 117 - 35
Candida and candidiasis: the cell wall as a potential molecular target for antifungal therapy; Gozalbo D et al.; The fungal species Candida albicans is an opportunistic pathogen, which causes serious infections in humans, particularly in immunocompromised patients . Depending on the underlying host defect, C . albicans causes a variety of infections, ranging from superficial mucocutaneous candidiasis to life-threatening disseminated infections . Both the limited spectrum of antifungal drugs currently in clinical use and the emergence of resistances make necessary the development of new effective antifungal drugs with minimal side effects; however, such a research is limited by the small number of specific target sites identified to date . The cell wall is a fungal specific dynamic structure essential to almost every aspect of the biology and pathogenicity of C . albicans . Its structure confers physical protection and shape to fungal cells, and as the most external part of the fungus, the cell wall mediates the interaction with the host, including adhesion to host tissues and modulation of the host anti-Candida immune response . Consequently, the fungal cell wall can be considered as a suitable target for development of new antifungal compounds . Therefore two distinct types of potential cell wall-related targets can be envisaged, according to their mode of action in inhibiting infection: (i) inhibition of cell wall biogenesis, which may impair cell wall integrity and thus cell viability, and (ii) modification of host-fungus interactions by inhibiting or blocking putative virulence factors, which may impair host colonization and progress of the infectious process . Antibodies specific to cell wall antigens may protect against infection by a variety of mechanisms and may evolve into save antifungal agents.

Mycopathologia, 2004 Apr, 157(3), 255 - 61
Prevalence and exoenzyme secretion by Candida albicans isolates from oral and vaginal mucosas of HIV-infected women; Ribeiro MA et al.; A cross-sectional study was performed to evaluate the prevalence and the aetiology of forms of mucosal fungal infections of HIV-negative and HIV-positive women . Candida albicans was the predominate specie isolated from both groups of patients, with remarkable proportion of isolation from symptomatic women . All 239 C . albicans isolates, regardless of their source, showed activity of proteinase and phospholipase . It was verified that isolates with particularly higher levels of exoenzymes production were significantly more common in HIV-positive patients . However, isolates obtained from the HIV-positive patients in use of HAART, with protease inhibitor, presented lower levels of these exoenzymes, similar to the levels observed in the isolates from HIV-negative patients.

Br J Neurosurg, 2004 Apr, 18(2), 189 - 92
Candida albicans spondylodiscitis; Ugarriza LF et al.; Candida spondylodiscitis is a rare complication of a haematogenous dissemination of a candida infection, that usually affects immunocompromised patients . We present a case of a Candida albicans spondylodiscitis in a patient in whom a bacterial origin was suspected because of the antecedent of a Staphylococcus aureus bacteriaemia . After unfavourable evolution with initial antibiotic treatment, the correct diagnosis was reached after culture of the material obtained from surgical debridement . The clinical, diagnostic features, and the literature are reviewed.

J Drugs Dermatol, 2004 May-Jun, 3(3), 263 - 5
Intralesional immunotherapy for warts using a combination of skin test antigens: a safe and effective therapy; Johnson SM et al.; Warts are a common dermatologic problem . There are many treatments available . Intralesional injection of a skin test antigen has been shown to be efficacious at eradicating all warts when only a part of one wart is injected . The aim of this study was to determine whether injection of the combination of Candida albicans, mumps, and Trichophyton skin test antigens was more efficacious than and as safe as single antigen injection . Seventy-one percent of subjects had resolution of their warts with the injection of the combination of skin test antigens in this open label single arm study.

Curr Microbiol, 2004 Jun, 48(6), 447 - 51
Relationship between expression of cell surface hydrophobicity protein 1 (CSH1p) and surface hydrophobicity properties of Candida dubliniensis; Hazen KC; Candida dubliniensis and Candida albicans cause most of the oral candidiasis infections in AIDS patients . Unlike C . albicans, which variably expresses cell surface hydrophobicity (CSH) depending on environmental conditions, C . dubliniensis is hydrophobic under all environmental conditions . C . dubliniensis produces CdCSH1p, a protein related to CaCSH1p that contributes to CSH expression of C . albicans . We investigated whether environmental conditions affect CdCSH1p expression, CSH avidity, and adhesion to fibronectin (Fn) . C . dubliniensis CD36 was grown at 23 degrees C and 37 degrees C in four different media . CdCSH1p expression was affected by growth temperature, with cells grown at 37 degrees C expressing the protein, but cells grown at 23 degrees C did not . Hydrophobic avidity for two media was higher in cells grown at 37 degrees C than at 23 degrees C . Cells grown at 23 degrees C were generally less adherent than 37 degrees C-grown cells to Fn . The results suggest CdCSH1p but not hydrophobic avidity may have a role in adhesion of C . dubliniensis to Fn.

Bratisl Lek Listy, 2003, 104(7-8), 211 - 7
Study of the dermatophytes in dogs and the risk of human infection; Kozak M et al.; BACKGROUND: The infiltrative growth of fungi that multiplies in the tissues (dermatophytes, candida, agents of visceral mycoses) is the generally known proof of "active" functioning of these organisms . The relative importance of fungi as inducers of human and animal diseases grows constantly . METHODS: The study was carried out on dogs that showed skin changes . The frequency of examinations was determined on the basis of anamneses . Skin scrapings and affected fragile or dull hair and skin swabs were subject to Laboratory diagnostic examinations based on cultivation and microscopical examination . Our evaluations were focused on the type of parasite--ecothrix, endothrix, neoendothrix, and the size of arthrospores . RESULTS: The results of our study are presented as the incidence of dermatophytes and other micromycetes in 100 samples of skin scrapings and swabs from 100 dogs of both sexes, different breeds and age categories that had exhibited skin problems . We isolated 12 species . Malassezia pachydermatis was the most frequently detected species and was isolated from 31 cases . Trichophyton mentagrophytes was isolated from 2 samples, species Microsporum canis was isolated only from one case, Candida albicans was isolated from 3 samples . The agent Candida krusei was isolated from 5 samples, Candida pulcherrima a relatively rare candidal species, was isolated from skin of an 8-year old Dalmatian bitch, Candida sp . in 5 cases . Trichosporon cutaneum was isolated from a 3-year old Laika etc . CONCLUSION: This study confirms the need of constant research in this area . This indicates that the incidence of mycoses in dogs and other pets is of importance in the exposure of people to the risk of acquiring mycotic infections . (Tab . 1, Fig . 5, Ref . 57).

Akush Ginekol (Sofiia), 2004, 43(1), 20 - 3
{Comparative study of itraconazole and fluconazole therapy in vaginal candidosis}; Vacheva-Dobrevski R et al.; Women who had symptomatic acute vulvovaginal candidiasis are include in this study . Micological investigation is realized by microscopy, culture method (CHROM agar, BD, USA) and ID 32 C (Fungus), Mini API Bio Merieux, France . Most frequent isolates are Candida albicans (82.67%), followed by C . glabrata (7.8%) and C . parapsilosis (5.51%) . We compare two groups of patients: received itraconazole (200 mg bd oral dose for 1 day) and fluconazole (150 mg single oral dose) . The rate of mycological cure is 93.8% in the itraconazole group and 79.03% in the fluconazole group (p = 0.008) . Clinical response rate for women receiving itraconazole (80%) is significantly higher than fluconazole group (59.7%) . These results suggests that itraconazole is more effective than fluconazole in the treatment of acute vulvovaginal candidiasis.

J Clin Pathol, 2004 Jun, 57(6), 598 - 603
Tissue invasiveness and non-acidic pH in human candidiasis correlate with "in vivo" expression by Candida albicans of the carbohydrate epitope recognised by new monoclonal antibody 1H4; Monteagudo C et al.; BACKGROUND: The morphogenetic conversion between yeast and hyphal growth forms appears to be crucial in the pathogenesis of invasive candidiasis, and can be regulated by environmental signals such as extracellular pH . AIMS: To characterise the epitope recognised by monoclonal antibody 1H4, and to evaluate the expression of its corresponding epitope in Candida albicans cells under different conditions of pH and temperature, and "in vivo", in tissue samples from patients with human candidiasis . METHODS: Monoclonal antibody 1H4 was generated against the 58 kDa cell wall mannoprotein of C albicans (mp58), and was further characterised by immunoblot analysis, periodate treatment of the antigenic preparations, and agglutination experiments of C albicans strains 3153A, SC5314, and 412, cultured under different environmental conditions (growth media and pH) . An immunohistochemical study was performed in 24 human tissue samples from patients with mucocutaneous and systemic candidiasis . RESULTS: 1H4 recognises a pH sensitive carbohydrate epitope on the surface of C albicans cells, and this epitope is not restricted to mp58, but is shared with other cell wall mannoproteins . Immunohistochemical findings indicated that expression of the 1H4 epitope on C albicans cells in tissue sections from human candidiasis correlates with tissue invasion and pH of the niche . 1H4 immunoreactivity was also found in candida remnants within macrophages . CONCLUSIONS: The fact that 1H4 epitope expression selectively identifies invasive forms of C albicans, in addition to candida remnants within macrophages, supports its potential value in the diagnosis and management of human candidiasis.

Mol Microbiol, 2004 Jun, 52(5), 1451 - 62
Homozygosity at the MTL locus in clinical strains of Candida albicans: karyotypic rearrangements and tetraploid formation; Legrand M et al.; One hundred and twenty Candida albicans clinical isolates from the late 1980s and early 1990s were examined for homozygosity at the MTL locus . Of these, 108 were heterozygous (MTLa/MTLalpha), whereas seven were MTLa and five were MTLalpha . Five of the homozygous isolates were able to switch to the opaque cell morphology, while opaque cells were not detectable among the remaining seven . Nevertheless, all but one of the isolates homozygous at the MTL locus were shown to mate and to yield cells containing markers from both parents; the non-mater was found to have a frameshift in the MTLalpha1 gene . In contrast to Saccharomyces cerevisiae, C . albicans homozygotes with no active MTL allele failed to mate rather than mating as a cells . There was no correlation between homozygosity and fluconazole resistance, mating and fluconazole resistance or switching and fluconazole resistance, in part because most of the strains were isolated before the widespread use of this antifungal agent, and only three were in fact drug resistant . Ten of the 12 homozygotes had rearranged karyotypes involving one or more homologue of chromosomes 4, 5, 6 and 7 . We suggest that karyotypic rearrangement, drug resistance and homozygosity come about as the result of induction of hyper-recombination during the infection process; hence, they tend to occur together, but each is the independent result of the same event . Furthermore, as clinical strains can mate and form tetraploids, mating and marker exchange are likely to be a significant part of the life cycle of C . albicans in vivo.

Pediatr Dermatol, 2004 May-Jun, 21(3), 260 - 1
Invasive fungal dermatitis in a 770 gram neonate; Passeron T et al.; A 770 g birthweight, 25-weeks gestation infant girl was born from a bigeminal pregnancy . Six days later she developed erythematous "diaper dermatitis" and maceration of the flexural areas . Despite topical antifungal therapy, erythematous plaques appeared 2 days later on the back . Within less than 24 hours, skin erosions with crusting appeared and spread over the whole body . Candida albicans was found in cutaneous scales, and blood, umbilical catheter, and cerebrospinal fluid cultures . In spite of intravenous fluconazole therapy, her general condition deteriorated and she died 2 days later . This premature neonate had a typical case of invasive fungal dermatitis, which is characterized by diffuse erosive and crusting cutaneous lesions appearing several days after birth and a high rate of systemic fungal infection (mainly but not exclusively) due to Candida sp . Mortality is high and prompt diagnosis and initiation of antifungal therapy appears to be the most important factor for survival.

Cytokine, 2004 Feb 21, 25(4), 162 - 71
In vitro analyses of tissue structure and interleukin-1beta expression and production by human oral mucosa in response to Candida albicans infections; Mostefaoui Y et al.; Clinical and experimental observations suggest that oral epithelial cells play a key role in host defenses against candidal infections through cytokines and chemokines . We thus attempted to determine whether oral epithelial cells convey IL-1beta as a pro-inflammatory cytokine in response to Candida albicans infections . We created engineered human oral mucosa (EHOM), put them in contact with live and heat-inactivated C . albicans (10(5) yeast/cm2), and measured the expression of IL-1beta mRNA and protein . Tissue structure and C . albicans morphology were also evaluated . Only live C . albicans modulated IL-1beta expression and secretion . IL-1beta mRNA expression significantly increased during the early stages of infection and decreased during the later stages . The modulatory effect of C . albicans on IL-1beta expression was confirmed by the fact that increased amounts of inactive IL-1beta (33 kDa) were detected early during the infection which then dropped dramatically . There was a significant and time-dependent increase in the amount of the active form of IL-1beta (17 kDa) secreted into the supernatant by epithelial cells infected with live C . albicans . Histological features revealed damage to infected tissues (separation of epithelial cells, edema, vacuolization, reduction in thickness) compared to uninfected ones . Morphological analyses showed that C . albicans changed from a blastospore to a hyphal form at later infection periods . This transformation was very pronounced at 8 and 24 h post-infection . These results provide additional evidence for the contribution of oral epithelial cells to local defenses against exogenous stimulations such as C . albicans infections.

Anticancer Res, 2004 Mar-Apr, 24(2B), 1025 - 30
Human beta-defensin-2 in oral cancer with opportunistic Candida infection; Meyer JE et al.; Candida albicans (CA) is a frequent opportunistic pathogen in cancer patients . Usually, human surfaces are protected, apart from physical barriers, by the production of human beta-defensins (hBD) . hBD-2 shows a potent antimicrobial activity against CA . We therefore investigated whether CA induces hBD-2 expression in primary oral cells and if immunosuppressive betamethasone alters hBD-2 expression . Additionally, we studied, whether a lack of hBD-2 expression could explain opportunistic infection of tonsillar cancer . Primary oral epithelial cells and fibroblasts were stimulated with Candida albicans in a time- and dose-dependent manner with or without betamethasone preincubation . Total RNA from oral cells and specimens was isolated and hBD-2 expression was analyzed by semiquantitative RT-PCR . Our data demonstrate that opportunistic CA induced hBD-2 expression in a time- and dose-dependent manner, suggesting hBD-2 to be a fast antifungal, epithelia-derived immune response . Treatment with glucocorticoid could lead to diminished innate immunity based on suppression of inducible AP . Malignant transformation induces alteration of hBD-2 expression and leads to a reduced hBD-2 expression and subsequentially to Candida colonization on oral SCCs.

Arch Biochem Biophys, 2004 Jun 15, 426(2), 148 - 56
Sensing the host environment: recognition of hemoglobin by the pathogenic yeast Candida albicans; Pendrak ML et al.; Adhesion to host cells and tissues is important for several steps in the pathogenesis of disseminated Candida albicans infections . Although such adhesion is evident in vivo and for C . albicans grown in vitro in complex medium, some adhesive activities are absent when cultures are grown in defined media . However, addition of hemoglobin to defined media restores binding and adhesion to several host proteins . This activity of hemoglobin is independent of iron acquisition and is mediated by a cell surface hemoglobin receptor . In addition to regulating expression of adhesion receptors, hemoglobin rapidly induces expression of several genes . One of these, a heme oxygenase, allows the pathogen to utilize exogenous heme or hemoglobin to acquire iron and to produce the cytoprotective molecules alpha-biliverdin and carbon monoxide . The specific recognition of and responses to hemoglobin demonstrate a unique adaptation of C . albicans to be both a commensal and an opportunistic pathogen in humans.

FEMS Microbiol Lett, 2004 Jun 1, 235(1), 169 - 76
Cloning and functional characterization of the Rhizopus oryzae high affinity iron permease (rFTR1) gene; Fu Y et al.; Rhizopus oryzae is the most common etiologic agent of mucormycosis . Clinical and animal model data clearly demonstrate that the presence of elevated available serum iron predisposes the host to develop mucormycosis . Therefore, the high affinity iron permease (rFTR1) which encodes a protein required to scavenge iron from the environment, is highly likely to be a critical determinant of virulence for R . oryzae . We have cloned rFTR1 by using a PCR approach relying on degenerate primers designed from the conserved regions of Saccharomyces cerevisiae high affinity iron permease . Sequence analysis of a 2.0 kb EcoRI genomic clone revealed a single open reading frame of 1107 bp that lacked introns . The putative rFtr1p had significant homology to known fungal high affinity iron permeases from Candida albicans (46% identity) and S . cerevisiae (44% identity) . In R . oryzae, rFTR1 was expressed in iron-depleted and not in iron-rich media . Finally, rFTR1 restored the ability of an ftr1 null mutant of S . cerevisiae to grow on iron-limited medium and to take up radiolabeled iron, whereas S . cerevisiae transformed with the empty vector did not . These data demonstrate that we have cloned the gene encoding a R . oryzae high affinity iron permease and the putative rFtr1p is involved in assimilation of iron from iron-depleted environments.

FEMS Microbiol Lett, 2004 Jun 1, 235(1), 73 - 80
Phosphorylation of glucosamine-6-phosphate synthase is important but not essential for germination and mycelial growth of Candida albicans; Gabriel I et al.; A site-directed mutagenesis of the GFA1 gene encoding Candida albicans glucosamine-6-phosphate (GlcN-6-P) synthase afforded its GFA1S208A version . A product of the modified gene, lacking the putative phosphorylation site for protein kinase A (PKA), exhibited all the basic properties identical to those of the wild-type enzyme but was no longer a substrate for PKA . Comparison of the C . albicans Deltagfa1/GFA1 and Deltagfa1/GFA1S208A cells, grown under conditions stimulating yeast-to-mycelia transformation, revealed that the latter demonstrated lower GlcN-6-P synthase specific activity, decreased chitin content and formed much fewer mycelial forms . All these findings, as well as the observed effects of specific inhibitors of protein kinases, suggest that a loss of the possibility of GlcN-6-P synthase phosphorylation by PKA strongly reduces but not completely eliminates the germinative response of C . albicans cells.

Microbes Infect, 2004 May, 6(6), 542 - 8
Toll-like receptor 2 is dispensable for acquired host immune resistance to Candida albicans in a murine model of disseminated candidiasis; Villamon E et al.; Previous work by our group showed that Toll-like receptor 2 (TLR2) is essential for activation of innate immunity, playing a major role in the response of macrophages to Candida albicans, triggering cytokine and chemokine expression, and therefore TLR2 -/- mice are more susceptible to systemic primary candidiasis . In this work, we used a murine model of systemic C . albicans infection, in which resistance to reinfection with virulent wild-type cells is induced by prior exposure of mice to a low-virulence agerminative strain of C . albicans (primary sublethal infection), to study the influence of TLR2 gene deletion on (i) the ability to develop an acquired resistance upon vaccination; (ii) the development of the acquired humoral response; and (iii) the production of Th1 cytokines IFN-gamma, IL-12 and TNF-alpha . Our results indicate that, although TLR2 -/- mice have a very impaired production of Th1 cytokines compared with control mice, they are equally capable of mounting a specific humoral response to the fungus and developing a vaccine-induced resistance.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2305 - 7
Enhanced production of farnesol by Candida albicans treated with four azoles; Hornby JM et al.; The dimorphic fungus Candida albicans excretes farnesol, which is produced enzymatically from the sterol biosynthetic intermediate farnesyl pyrophosphate . Inhibition of C . albicans by four azole antifungals, fluconazole, ketoconazole, miconazole, and clotrimazole, caused elevated farnesol production (10- to 45-fold) . Furthermore, farnesol production occurs in both laboratory strains and clinical isolates (J . M . Hornby et al., Appl . Environ . Microbiol . 67:2982-2992, 2001) of C . albicans.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2223 - 7
Clade-specific flucytosine resistance is due to a single nucleotide change in the FUR1 gene of Candida albicans; Dodgson AR et al.; Population studies have indicated that natural resistance to flucytosine (5FC) in Candida albicans is limited to one of the five major clades, clade I . In addition, while 73% of clade I isolates are less susceptible to 5FC (MIC >/= 0.5 microg/ml), only 2% of non-clade I isolates are less susceptible . In order to determine the genetic basis for this clade-specific resistance, we sequenced two genes involved in the metabolism of 5FC that had previously been linked to resistance (cytosine deaminase and uracil phosphoribosyltransferase), in 48 isolates representative of all clades . Our results demonstrate that a single nucleotide change from cytosine to thymine at position 301 in the uracil phosphoribosyltransferase gene (FUR1) of C . albicans is responsible for 5FC resistance . The mutant allele was found only in group I isolates . The 5FC MICs for strains without copies of the mutant allele were almost exclusively </=0.25 microg/ml, those for strains with one copy of the mutant allele were >/=0.5 microg/ml, and those for strains with two copies of the mutant allele were >/=16 microg/ml . Thus, the two alleles were codominant . The presence of this allele is responsible for clade I-specific resistance to 5FC within the C . albicans population and thus by inference is likely to be the major underlying 5FC resistance mechanism in C . albicans . This represents the first description of the genetic mutation responsible for 5FC resistance.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2124 - 31
Application of real-time quantitative PCR to molecular analysis of Candida albicans strains exhibiting reduced susceptibility to azoles; Chau AS et al.; Real-time quantitative PCR was used to measure expression levels of genes encoding efflux pumps, ERG11 and two control genes, ACT1 and PMA1, in a collection of 14 fluconazole-susceptible Candida albicans isolates . For each gene, average expression levels and variations within the population were determined . These values were then used as reference points to make predictions about the molecular basis of resistance in 38 clinical isolates (the majority of which were resistant to fluconazole) obtained from 18 patients treated with posaconazole for refractory oropharyngeal candidiasis . For each of the 38 isolates, the expression levels of genes encoding efflux pumps, ERG11 and the control genes, were measured as above . Comparison of the two data sets revealed that expression of ACT1 and PMA1 did not vary significantly between the two sets of isolates . In contrast, MDR1, ERG11, CDR1, and CDR2 were overexpressed in 3, 4, 14, and 35, respectively, of the isolates from patients treated with azoles . In addition to these changes, the patient isolates all had at least one and often multiple missense mutations in ERG11 . Select ERG11 alleles were expressed in Saccharomyces cerevisiae; all of the alleles tested conferred reduced susceptibility to fluconazole . Despite both the increases in pump expression and the ERG11 mutations, only one of the patient isolates exhibited a large decrease in posaconazole susceptibility.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 1974 - 82
Anti-inflammatory effects of moxifloxacin on activated human monocytic cells: inhibition of NF-kappaB and mitogen-activated protein kinase activation and of synthesis of proinflammatory cytokines; Weiss T et al.; We previously showed that moxifloxacin (MXF) exerts protective anti-inflammatory effects in immunosuppressed mice infected with Candida albicans by inhibiting interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) production in the lung . Immunohistochemistry demonstrated inhibition of nuclear factor (NF)-kappaB translocation in lung epithelium and macrophages in MXF-treated mice . In the present study we investigated the effects of MXF on the production of proinflammatory cytokines (i.e., IL-8, TNF-alpha, and IL-1beta) by activated human peripheral blood monocytes and THP-1 cells and analyzed the effects of the drug on the major signal transduction pathways associated with inflammation: NF-kappaB and the mitogen-activated protein kinases ERK and c-Jun N-terminal kinase (JNK) . The levels of IL-8, TNF-alpha, and IL-1beta secretion rose 20- and 6.7-fold in lipopolysaccharide (LPS)-activated monocytes and THP-1 cells, respectively . MXF (5 to 20 microg/ml) significantly inhibited cytokine production by 14 to 80% and 15 to 73% in monocytes and THP-1 cells, respectively . In THP-1 cells, the level of NF-kappaB nuclear translocation increased fourfold following stimulation with LPS-phorbol myristate acetate (PMA), and this was inhibited (38%) by 10 microg of MXF per ml . We then assayed the degradation of inhibitor (I)-kappaB by Western blotting . LPS-PMA induced degradation of I-kappaB by 73%, while addition of MXF (5 microg/ml) inhibited I-kappaB degradation by 49% . Activation of ERK1/2 and the 46-kDa p-JNK protein was enhanced by LPS and LPS-PMA and was significantly inhibited by MXF (54 and 42%, respectively, with MXF at 10 microg/ml) . We conclude that MXF suppresses the secretion of proinflammatory cytokines in human monocytes and THP-1 cells and that it exerts its anti-inflammatory effects in THP-1 cells by inhibiting NF-kappaB, ERK, and JNK activation . Its anti-inflammatory properties should be further assessed in clinical settings.

Anticancer Res, 2004 Mar-Apr, 24(2A), 405 - 8
Adriamycin alters the expression of drug efflux pumps and confers amphotericin B tolerance in Candida albicans; O'Keeffe J et al.; The aim of the work presented here was to establish whether exposure of the yeast C . albicans to adriamycin altered the expression of the CDR1 drug efflux pump and consequently altered the susceptibility of the yeast to amphotericin B . Using a monoclonal antibody directed against human MDR1 and polyclonal antibodies against CDR1 (Candida drug resistance), we demonstrated that adriamycin induces an elevation in the expression of the CDR1 efflux pump which, together with previously recorded alterations in the composition of the fungal cell membrane, may confer tolerance to amphotericin B . This work highlights the fact that adriamycin therapy may inadvertently alter the susceptibility of C . albicans to amphotericin B, which may have deleterious consequences for anti-cancer chemotherapy regimes incorporating this anti-neoplastic agent.

Reprod Biomed Online, 2004 May, 8(5), 569 - 73
Inhibition of oocyte fertilization by assisted reproductive techniques and increased sperm DNA fragmentation in the presence of Candida albicans: a case report; Burrello N et al.; The effects of Candida albicans on sperm parameters and the outcome of infertility treatment are unclear . This report describes a lack of fertilization after assisted reproductive techniques and increased sperm DNA fragmentation in an infertile patient with male accessory gland infection due to Candida albicans . He had normal sperm parameters and, therefore, underwent conventional IVF for a female factor of infertility . No spermatozoa or only one spermatozoon per oocyte were found attached to the zona pellucida of the six mature oocytes retrieved . A new semen sample was then requested from the patient to perform intracytoplasmic sperm injection (ICSI) on the same oocytes, but again no fertilization resulted . Candida albicans was detected in the medium where spermatozoa were co-incubated with oocytes and subsequently in the urethral swabs . It did not have any detrimental effect on sperm parameters soon after ejaculation or following separation of motile spermatozoa by swim-up technique . Fertilization failure after assisted reproduction treatment was associated with an increased percentage of motile spermatozoa having chromatin packaging abnormalities, externalization of phosphatidylserine and DNA fragmentation . In conclusion, Candida albicans did not affect sperm parameters, but increased sperm chromatin packaging damage and apoptosis that might have caused fertilization failure after assisted reproduction treatment in this couple.

J Med Microbiol, 2004 Jun, 53(Pt 6), 495 - 500
Effect of orally administered bovine lactoferrin on the immune response in the oral candidiasis murine model; Takakura N et al.; Therapeutic activity against oral candidiasis of orally administered bovine lactoferrin (LF), a multifunctional milk protein, was shown in a previous report using an immunosuppressed murine model . In the present study, the influence of orally administered LF on immune responses relevant to this therapeutic effect was examined . Because mice were immunosuppressed with prednisolone 1 day before and 3 days after the infection with Candida, the numbers of peripheral blood leukocytes (PBL) and cervical lymph node (CLN) cells were reduced . LF feeding prevented the reduction in the numbers of PBL on day 1 and CLN cells on days 1, 5 and 6 in the Candida-infected mice . The number of CLN cells of individual mice on days 5 and 6 was inversely correlated with the Candida c.f.u . in the oral cavity . Increased production of IFN-gamma and TNF-alpha by CLN cells stimulated with heat-killed Candida albicans on day 6 was observed in LF-treated mice compared with non-treated mice . Concanavalin A (ConA)-stimulated CLN cells from LF-treated mice also showed a significant increase in the production of IFN-gamma and IL12 on day 5 and a tendency for increased production of IFN-gamma and TNF-alpha on day 6 . The levels of cytokine production by ConA-stimulated CLN cells on day 6 were inversely correlated with the Candida c.f.u . in the oral cavity . In conclusion, the alleviation of oral candidiasis by LF feeding in this model may correlate with the enhancement of the number of leukocytes and their cytokine responses in regional lymph nodes against Candida infection.

J Inorg Biochem, 2004 Jun, 98(6), 1023 - 31
Synthesis, antimicrobial activity and chemotherapeutic potential of inorganic derivatives of 2-(4'-thiazolyl)benzimidazole{thiabendazole}: X-ray crystal structures of {Cu(TBZH)2Cl}Cl.H2O.EtOH and TBZH2NO3 (TBZH=thiabendazole); Devereux M et al.; Thiabendazole (TBZH) reacts with iron(III) nitrate causing protonation of the ligand to yield the nitrate salt {TBZH(2)NO(3)} (1) . Reaction of TBZH with copper(II) acetate results in the deprotonation of the ligand yielding {Cu(TBZ)2.(H2O)2} (2) . Reactions of TBZH with the chloride, nitrate and butanedioate salts of copper(II) yields {Cu(TBZH)2Cl}Cl.H2O.EtOH (3), {Cu(TBZH)(2)(NO(3))(2)} (4) and {Cu(TBZH)(O(2)C-CH(2)CH(2)-CO(2))} (5), respectively . The TBZH acts as a neutral chelating ligand in 3-5 . Molecular structures of 1 and 3 were determined crystallographically . In 1, the asymmetric unit contains one TBZH(2)(+) cation and one NO(3)(-) anion . The structure of 3 comprises a five coordinate copper centre with the metal bound to two chelating TBZH ligands and one chloride . The geometry is best described as trigonal bipyramidal . Hydrogen bonding connects the complex cation with the uncoordinated chloride anion and the water and ethanol solvate molecules . Compound 1 and the copper complexes 2-5, the metal free ligands and a number of simple copper(II) salts were each tested for their ability to inhibit the growth of Candida albicans . The metal free TBZH and its nitrate salt (1) exhibited very poor activity . Complex 2, in which the TBZH is present as an anionic ligand (TBZ(-)), exhibits moderate activity towards the pathogen . Chelation of the neutral TBZH to copper centres (complexes 3-5) results in potent anti-candida activity . The dimethyl sulphoxide (DMSO) soluble complexes 3 and 4, along with metal free TBZH were assessed for their cancer chemotherapeutic potential towards two human epithelial-derived cancer model cell lines . Complexes 3 and 4 displayed similar dose-dependent cytotoxicity in both cell lines with IC(50) values of approximately 50 microM, which were found to be significantly lower than that for metal free TBZH.

J Comp Physiol {B}, 2004 Jul, 174(5), 421 - 7 Epub 2004 May 18.
Comparative study of the heterophil phagocytic function in young and old ring doves (Streptopelia risoria) and its relationship with melatonin levels; Terron MP et al.; A functional connection between the pineal gland (via the hormone melatonin) and the immune system has been suggested . In our previous results in the ring dove, we observed diurnal oscillations in the levels of this neurohormone in young animals and a decline in its plasma levels with advancing age (which is accompanied by the absence of diurnal rhythm) . We also noted enhanced phagocytic activity of heterophils from old animals after in vitro incubation with both physiological and pharmacological doses of melatonin . Here, we evaluate the functional capacity of ring dove (Streptopelia risoria) heterophils in young (2 years of age) and old (8 years and more) animals at different times of day (0:00, 10:00 and 16:00, the times when the maximum, minimum, and mean values, respectively, of melatonin levels are observed in young animals) . The phagocytic capacities for the ingestion of latex beads and Candida albicans were evaluated, as well as the oxidative metabolism which accompanies phagocytosis . At all three times of day studied, the heterophil phagocytic function with both latex and C . albicans was significantly greater in the young than in the old animals, and in the young animal cells it was significantly higher at 0:00 . In addition, in the presence of latex beads, there was a significant decline at 10:00 and 0:00 of superoxide anion levels in the young animals relative to the old . In the young animals, there was a decline at 0:00 in comparison with both 10:00 and 16:00, and in the old animals there was a decline at both 0:00 and 16:00 compared with 10:00 . These results could be due, at least in part, to the absence of a diurnal rhythm of melatonin in old animals, and to an enhancing effect of that hormone on young animals' heterophil phagocytic function, which would also neutralize the oxidative stress deriving from this immune function.

J Antimicrob Chemother, 2004 Jun, 53(6), 1081 - 5 Epub 2004 May 12.
Antifungal effects of Melaleuca alternifolia (tea tree) oil and its components on Candida albicans, Candida glabrata and Saccharomyces cerevisiae; Hammer KA et al.; OBJECTIVES: The aim of this study was to investigate the mechanism of action of tea tree oil and its components against Candida albicans, Candida glabrata and Saccharomyces cerevisiae . METHODS: Yeast cells were treated with tea tree oil or components, at one or more concentrations, for up to 6 h . During this time, alterations in permeability were assessed by measuring the leakage of 260 nm absorbing materials and by the uptake of Methylene Blue dye . Membrane fluidity was measured by 1,6-diphenyl-1,3,5-hexatriene fluorescence . The effects of tea tree oil on glucose-induced medium acidification were quantified by measuring the pH of cell suspensions in the presence of both tea tree oil and glucose . RESULTS: The treatment of C . albicans with tea tree oil and components at concentrations of between 0.25 and 1.0% (v/v) altered both permeability and membrane fluidity . Membrane fluidity was also increased when C . albicans was cultured for 24 h with 0.016%-0.06% (v/v) tea tree oil, as compared with control cells . For all three organisms, glucose-induced acidification of the external medium was inhibited in a dose-dependent manner in the presence of 0.2%, 0.3% and 0.4% tea tree oil . CONCLUSIONS: Data from this study support the hypothesis that tea tree oil and components exert their antifungal actions by altering membrane properties and compromising membrane-associated functions.

J Oral Rehabil, 2004 May, 31(5), 460 - 7
An in vitro study into the effect of a limited range of denture cleaners on surface roughness and removal of Candida albicans from conventional heat-cured acrylic resin denture base material; Harrison Z et al.; This study evaluated the abrasiveness of four denture cleaners on the surface of denture base material and assessed their ability to remove Candida albicans . Acrylic resin discs 20 mm diameter and 2 mm thick were identically produced and polished . Four cleaners were evaluated: conventional toothpaste; toothpaste with stain remover; denture cleaning paste and an immersion type cleaner, and water were used as control . These were used at dilutions of 1:1, 1:2 and 1:3 with water . An electric toothbrush was used, and the discs cleaned to simulate 1 years' cleaning . The surface roughness of the discs were then measured, before and after cleaning, using a stylus profilometer, then inoculated with 1.2 x 10(6)C . albicans cells . The effectiveness of the denture cleaners to remove C . albicans cells was assessed following a single cleaning event . The immersion cleaner was significantly less abrasive than paste cleaners (P < 0.05) . There were no significant differences between any dilutions for any cleaner used (P > 0.05) . Immersion and paste cleaners removed almost all recoverable C . albicans from the discs, as cleaning with water alone was less effective (P < 0.05) . An immersion type cleaner was found to be the most suitable cleaner because of its low abrasivity and effective removal of organic debris.

Biol Pharm Bull, 2004 May, 27(5), 751 - 2
Farnesol, a morphogenetic autoregulatory substance in the dimorphic fungus Candida albicans, inhibits hyphae growth through suppression of a mitogen-activated protein kinase cascade; Sato T et al.; Candida albicans grew in hyphal form in RPMI1640, however, addition of farnesol inhibited the formation . Farnesol did not affect the expression of mRNAs related to cAMP-EFG1 pathways . The mRNAs (HST7 and CPH1) in mitogen activated protein kinase (MAP) cascades were decreased in farnesol-treated cells, but CST20 was not . Furthermore, expression of general amino acid permease 1 (GAP1) was decreased by farnesol . We concluded that farnesol inhibits a MAP kinase cascades, and the suppression is a cause of interruption of hyphae formation.

Biol Pharm Bull, 2004 May, 27(5), 674 - 8
Role of polymorphonuclear leukocytes in the resistance of tumor-bearing mice against Candida albicans infection; Okawa Y et al.; Tumor-bearing mice showed a significant resistance against Candida albicans intravenous infection . Longer survival was observed in groups of mice inoculated with fungal cells 2-3 weeks after tumor transplantation with allogeneic sarcoma 180, syngeneic methylcholanthrene-induced Meth A fibrosarcoma, and MM 46 mammary carcinoma than in non-tumor-bearing mice inoculated only with fungal cells . This effect was not observed when the mice were infected only 1 week after tumor transplantation . A significant decrease in the number of C . albicans cells in the kidneys was observed in mice inoculated with fungal cells 2-3 weeks after tumor transplantation . In the tumor-bearing mice treated with cyclophosphamide (CY), a remarkable decrease in both the number of peripheral blood polymorphonuclear leukocytes (PMNs) and the defense against challenge with C . albicans cells was observed, as compared with the CY-untreated groups (normal and tumor-bearing mice) . A marked increase in the calcium concentration in serum and number, candidacidal activity, active oxygen level, and myeloperoxidase activity of PMNs was observed in the 2-3-week tumor-bearing mice . From these results, it is suggested that PMNs, which accumulated in the 2-3-week tumor-bearing mice, play an important role in the protection from C . albicans infection by increasing the number and the types of killing factors.

J Clin Microbiol, 2004 May, 42(5), 2275 - 8
Detection of Candida albicans mRNA in archival histopathology samples by reverse transcription-PCR; Beggs KT et al.; The feasibility of detecting Candida albicans mRNA in formalin-fixed paraffin-embedded archival human histopathology specimens by reverse transcription-PCR (RT-PCR) was investigated . RT with gene-specific primers was used to detect five single-copy C . albicans gene transcripts, including those of two housekeeping genes, in oral candidiasis samples up to 8 years of age.

J Clin Microbiol, 2004 May, 42(5), 2249 - 51
Collaborative study of the NCCLS and flow cytometry methods for antifungal susceptibility testing of Candida albicans; Chaturvedi V et al.; One hundred clinical isolates of Candida albicans were tested for amphotericin B and fluconazole susceptibilities by the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution test at center 1 (C1) . The same isolates were tested blinded at center 2 (C2) by NCCLS and flow cytometry (FC) methods . The agreement between NCCLS and FC methods ranged from 96 to 99%.

J Clin Microbiol, 2004 May, 42(5), 2085 - 93
Rapid quantification of drug resistance gene expression in Candida albicans by reverse transcriptase LightCycler PCR and fluorescent probe hybridization; Frade JP et al.; We developed a rapid, sensitive, and reproducible assay to quantify Candida albicans ACT1, CDR1, CDR2, ERG11, and MDR1 mRNA using a two-step reverse transcription and LightCycler real-time PCR (RT-LightCycler PCR) method with sequence-specific hybridization probes . We compared RT-LightCycler PCR with Northern hybridization for quantitative analysis of gene expression in isolates with various fluconazole susceptibilities . Specificity of each LightCycler PCR was verified by LightCycler melting curve analysis and agarose gel electrophoresis of amplified products . Correlation of quantification results between RT-LightCycler PCR and Northern hybridization yielded correlation coefficients of > or = 0.91 for all genes except MDR1 (0.74) . In this case, reduced correlation was due to the inability of Northern hybridization to accurately quantify the high MDR1 expression in a susceptible dose-dependent isolate which was shown by RT-LightCycler PCR to overexpress MDR1 >200-fold relative to the other isolates tested . In four isolates, low levels of CDR2 mRNA were detected by RT-LightCycler PCR but were undetectable by Northern hybridization . mRNA quantification by RT-LightCycler PCR correlates with Northern hybridization and offers additional advantages, including increased sensitivity and speed of analysis, along with lower RNA concentration requirements and an increased dynamic range of signal detection.

Ophthalmic Physiol Opt, 2004 May, 24(3), 218 - 24
Effect of storage temperatures and time on the efficacy of multipurpose solutions for contact lenses; Leung P et al.; PURPOSE: To determine the effect of storage time and temperature on the efficacy of four multipurpose solutions for soft contact lenses . METHOD: Aliquots of multipurpose solutions (OPTI-FREE Express, ReNu MultiPlus, COMPLETE and SOLO-care) stored at different temperatures over a 3-month period, were challenged with contact lens-related ocular pathogens, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans . RESULTS: The results showed that OPTI-FREE Express had the best activity against Ps . aeruginosa at all temperatures; ReNu MultiPlus performed well at 25 degrees C; COMPLETE barely achieved activity requirements at all temperatures, and lost efficacy after 2 months . SOLO-care maintained its activity best against Ps . aeruginosa at 30 degrees C . Storage at fridge temperature reduced activity of all solutions . Regardless of storage temperature, activities of all solutions against S . aureus markedly decreased by 2 months . Only OPTI-FREE Express met FDA requirements against C . albicans . CONCLUSION: Performance of multipurpose solutions is affected by time and temperature of storage . Contact lens users should be aware that the efficacy of opened solutions may not be sustained for as long as 3 months . Manufacturers should reconsider their recommendations to further safeguard the ocular health of contact lens wearers.

J Biol Chem, 2004 Jul 16, 279(29), 30480 - 9 Epub 2004 May 05.
Functional and structural diversity in the Als protein family of Candida albicans; Sheppard DC et al.; The human fungal pathogen Candida albicans colonizes and invades a wide range of host tissues . Adherence to host constituents plays an important role in this process . Two members of the C . albicans Als protein family (Als1p and Als5p) have been found to mediate adherence; however, the functions of other members of this family are unknown . In this study, members of the ALS gene family were cloned and expressed in Saccharomyces cerevisiae to characterize their individual functions . Distinct Als proteins conferred distinct adherence profiles to diverse host substrates . Using chimeric Als5p-Als6p constructs, the regions mediating substrate-specific adherence were localized to the N-terminal domains in Als proteins . Interestingly, a subset of Als proteins also mediated endothelial cell invasion, a previously unknown function of this family . Consistent with these results, homology modeling revealed that Als members contain anti-parallel beta-sheet motifs interposed by extended regions, homologous to adhesins or invasins of the immunoglobulin superfamily . This finding was confirmed using circular dichroism and Fourier transform infrared spectrometric analysis of the N-terminal domain of Als1p . Specific regions of amino acid hypervariability were found among the N-terminal domains of Als proteins, and energy-based models predicted similarities and differences in the N-terminal domains that probably govern the diverse function of Als family members . Collectively, these results indicate that the structural and functional diversity within the Als family provides C . albicans with an array of cell wall proteins capable of recognizing and interacting with a wide range of host constituents during infection.

Glycobiology, 2004 Jul, 14(7), 593 - 8 Epub 2004 May 05.
Hydrolysis of Man9GlcNAc2 and Man8GlcNAc2 oligosaccharides by a purified alpha-mannosidase from Candida albicans; Mora-Montes HM et al.; A soluble alpha-mannosidase from Candida albicans was purified to homogeneity by sequential size exclusion, ion exchange, and affinity chromatographies in columns of Sepharose CL6B, DEAE Bio-Gel A, and Concanavalin A Sepharose 4B, respectively . Analytical electrophoresis of the purified preparation in 10% SDS-polyacrylamide gels stained with Coomassie blue revealed a single polypeptide of 43 kDa that was responsible for enzyme activity . The purified enzyme primarily trimmed Man(9)GlcNAc(2) to produce Man(8)GlcNAc(2) isomer B and mannose as a function of time of incubation up to 12 h at 37 degrees C . Prolonged incubation with the enzyme resulted in the accumulation after 24 h of other oligosaccharides corresponding to Man(7)GlcNAc(2) and probably Man(6)GlcNAc(2) . These two products were also observed when Man(8)GlcNAc(2) isomer B instead of Man(9)GlcNAc(2) was used as substrate . Other oligosaccharides, such as Man(6)GlcNAc(2)-Asn, Man(5)GlcNAc(2)-Asn, and the alpha1,3- and alpha1,6-linked mannobiosides, were not hydrolyzed at all . These properties are consistent with an alpha1,2-mannosidase that may represent a new member of the glycosylhydrolase family 47.

Am J Ophthalmol, 2004 May, 137(5), 820 - 5
In vitro investigation of voriconazole susceptibility for keratitis and endophthalmitis fungal pathogens; Marangon FB et al.; PURPOSE: To update the spectrum of ocular fungal isolates and investigate the in vitro efficacy of voriconazole and other antifungals . DESIGN: Experimental study . METHODS: Microbiology database was scanned and fungal isolates associated with keratitis (419) and endophthalmitis (122) were analyzed for classification and isolate frequency . The Sensititre YeastOne microdilution antifungal susceptibility test was used to evaluate susceptibility (MICs) of 34 common fungal pathogens against amphotericin B, fluconazole, ketoconazole, 5-flucytosine, itraconazole, and voriconazole . Ten of the test isolates were sent to a reference laboratory to validate the Sensititre results . RESULTS: Fusarium species remains the most frequent corneal fungal pathogen (60.1%) . Colletotrichum species (4.1%) has emerged as the fifth most common mold in keratitis . Top yeast isolates from cornea included Candida albicans (52.3%) and Candida parapsilosis (37.3%) . Half of the intraocular pathogens were Candida species . Paecilomyces (2.9%) and Philophora (1.9) were unusual pathogens . In vitro susceptibility profiles were voriconazole (100%), ketoconazole (82.4%), amphotericin (76.5%), itraconazole (67%), fluconazole (60%), and 5-FC (60%) . Voriconazole MIC(90) were lowest for Candida species (0.016 microg/ml) and highest for Fusarium species (2 microg/ml) . Reference laboratory MICs correlated 100% for yeast isolates (0.016 microg/ml) but were fourfold higher for Fusarium species (8 microg/ml) . MIC(90) for Aspergillus species was 0.5 microg/ml . CONCLUSIONS: Candida, Fusarium, and Aspergillus species remain frequent fungal pathogens . Voriconazole may have a role in the therapeutic management of Candida and Aspergillus ocular infections . Clinical efficacy must determine the role for other fungal pathogens . Human use and animal models will determine its use in the clinical setting.

Proc Natl Acad Sci U S A, 2004 May 11, 101(19), 7287 - 92 Epub 2004 May 03.
Incorporating chemical modification constraints into a dynamic programming algorithm for prediction of RNA secondary structure; Mathews DH et al.; A dynamic programming algorithm for prediction of RNA secondary structure has been revised to accommodate folding constraints determined by chemical modification and to include free energy increments for coaxial stacking of helices when they are either adjacent or separated by a single mismatch . Furthermore, free energy parameters are revised to account for recent experimental results for terminal mismatches and hairpin, bulge, internal, and multibranch loops . To demonstrate the applicability of this method, in vivo modification was performed on 5S rRNA in both Escherichia coli and Candida albicans with 1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide metho-p-toluene sulfonate, dimethyl sulfate, and kethoxal . The percentage of known base pairs in the predicted structure increased from 26.3% to 86.8% for the E . coli sequence by using modification constraints . For C . albicans, the accuracy remained 87.5% both with and without modification data . On average, for these sequences and a set of 14 sequences with known secondary structure and chemical modification data taken from the literature, accuracy improves from 67% to 76% . This enhancement primarily reflects improvement for three sequences that are predicted with <40% accuracy on the basis of energetics alone . For these sequences, inclusion of chemical modification constraints improves the average accuracy from 28% to 78% . For the 11 sequences with <6% pseudoknotted base pairs, structures predicted with constraints from chemical modification contain on average 84% of known canonical base pairs.

Proc Natl Acad Sci U S A, 2004 May 11, 101(19), 7329 - 34 Epub 2004 May 03.
The diploid genome sequence of Candida albicans; Jones T et al.; We present the diploid genome sequence of the fungal pathogen Candida albicans . Because C . albicans has no known haploid or homozygous form, sequencing was performed as a whole-genome shotgun of the heterozygous diploid genome in strain SC5314, a clinical isolate that is the parent of strains widely used for molecular analysis . We developed computational methods to assemble a diploid genome sequence in good agreement with available physical mapping data . We provide a whole-genome description of heterozygosity in the organism . Comparative genomic analyses provide important clues about the evolution of the species and its mechanisms of pathogenesis.

Photochem Photobiol Sci, 2004 May, 3(5), 419 - 22 Epub 2004 Jan 28.
In vitro effect of 5-aminolaevulinic acid plus visible light on Candida albicans; Monfrecola G et al.; Photodynamic therapy, currently used as an alternative technique for the treatment of superficial non-melanoma skin cancers, has been employed in vitro to kill different species of microorganisms . Here the development of Candida albicans colonies has been measured after application of 5-aminolaevulinic acid (ALA) plus visible light (VIS) irradiation . C . albicans suspensions (10 colony forming units microl(-1)) have been prepared . For the experiment 30 microl of suspension have been incubated in the dark for 3 h, with increasing concentrations of ALA (125, 250, 300, 350, 400, 450, 500, 550, 600, 750, 1000 mg ml(-1)) and then irradiated with a fixed dose (40 J cm(-2)) of VIS . Immediately after the irradiative session, the C . albicans suspensions were disseminated on dishes containing a Sabouraud agar + CAF medium and cultured in the dark at 27 degree C; after 48 h colony development has been measured . In the same way four controls have been prepared: (i)C . albicans suspensions not treated with ALA-PDT; (ii)C . albicans suspensions incubated with increasing ALA concentrations without VIS; (iii)C . albicans suspensions irradiated with 40 J cm(-2) of VIS without ALA; (iv)C . albicans suspensions irradiated immediately after the addition of increasing concentrations of ALA without the 3 h incubation . Colonies treated with ALA-PDT have been studied with electron microscopy (E.M.) . It was found that: (i) none of the controls prepared modified the development of C . albicans colonies; (ii) ALA plus VIS inhibited C . albicans growth in a concentration-dependent way: up to 250 mg ml of ALA concentrations did not affect C . albicans cells, 300 mg ml(-1) induced a 50% reduction in the number of colonies, a complete inhibition started from concentrations of 600 mg ml(-1); (iii) after ALA-PDT E.M . showed modifications of the cell membranes . From the results it is concluded ALA plus VIS light is able to kill C . albicans colonies, at least in vitro . Although other pharmacological approaches are available, further studies could show that PDT is a potential treatment for candidosis.

Int J Antimicrob Agents, 2004 May, 23(5), 520 - 3
Differential effects of the combination of caspofungin and terbinafine against Candida albicans, Candida dubliniensis and Candida kefyr; Gil-Lamaignere C et al.; The activity of caspofungin (CSP) combined with terbinafine (TRB) against Candida dubliniensis, Candida kefyr and azole-resistant Candida albicans was evaluated in vitro by checkerboard analysis . The combination of CSP with TRB resulted in positive interactive effects in vitro against C . albicans and C . kefyr but not against C . dubliniensis . Moreover, true synergism was observed only against TRB resistant strains which became susceptible to this drug in the presence of CSP . In contrast, indifference was observed against strains that were already sensitive to TRB indicating that CSP may inhibit resistance to TRB.

J Ethnopharmacol, 2004 Apr, 91(2-3), 181 - 8
Phyllanthus piscatorum, ethnopharmacological studies on a women's medicinal plant of the Yanomamï Amerindians; Gertsch J et al.; The shrub Phyllanthus piscatorum Kunth (Euphorbiaceae) is cultivated by various ethnic groups of the Amazon because of its piscicidal properties . During ethnobotanical fieldwork among the Yanomami Amerindians in Venezuela we observed that Phyllanthus piscatorum was exclusively cultivated and used by the women . Aerial parts of this herbaceous shrub are employed as fish poison and medicine to treat wounds and fungal infections . In addition, the leaves are used as tobacco substitute . Ethnobotanical data regarding the context of the use of this plant are presented . To validate ethnobotanical information related to its medicinal indications, antimicrobial, and antiprotozoal properties of water, methanol (MeOH) and dichloromethane (DCM) extracts were studied . No activity against Gram-positive bacterial strains but significant activity against the fungi Aspergillus fumigatus, Aspergillus flavus and the yeast Candida albicans were found . All extracts showed weak in vitro activity against Plasmodium falciparum and Trypanosoma brucei rhodesiense . The extracts were further investigated for cytotoxic effects in an in vitro test system with leukemia Jurkat T, HeLa, and human peripheral mononuclear blood cells (PBMCs) . During the first 48 h the extracts did not exhibit any cytotoxicity . After 72 h the DCM extract potently inhibited viability of HeLa cells . Although in several communities along the upper Orinoco the cultivation and use of Phyllanthus piscatorum is being lost because of the ongoing acculturation, the traditional medicinal use of Phyllanthus piscatorum might provide an effective and cheap remedy against dermatological diseases linked with Candida albicans infections.

Quintessence Int, 2004 Mar, 35(3), 194 - 9
Evaluation of the sanitization effectiveness of a denture-cleaning product on dentures contaminated with known microbial flora . An in vitro study; Glass RT et al.; OBJECTIVES: To see if dentures contaminated with Staphylococcus aureus, Pseudomas aeruginosa, Bacillus cereus, Candida albicans, and herpes simplex virus 1 could be effectively decontaminated by using Medical Tabs for Dentures . METHOD AND MATERIALS: Ten methylmethacrylate dentures with processed soft liners (soft-liner dentures) and 10 methylmethacrylate dentures without processed soft liners (hard dentures) were aseptically fragmented and individually incubated with a target microorganism . Test denture fragments were immersed in Medical for 5 minutes, vortexed for 5 minutes, and serially diluted onto media . The control denture fragments were similarly treated in sterile water . For virus contamination, denture fragments were contaminated with 1.2 x 10(9) tissue culture infective dose (TCID)50/mL . They were treated with either Medical for 5 minutes (test fragments) or water (controls) for 5 minutes . Serial dilutions were performed and viral (TCID)50/mL titers were calculated using the Reed-Muench method . RESULTS: Medical treatments effectively eliminated C . albicans, S . aureus, and P . aeruginosa from soft-liner dentures . Treatment of hard dentures eradicated C . albicans and reduced the numbers of S . aureus and P . aeruginosa to < 10 . B . cereus showed a reduction of 10 microorganisms in hard dentures while the soft-liner dentures did not show an appreciable reduction . Viral analyses found that both types of dentures retained large amounts of virus when washed with water, but no virus was recovered from any of the 40 samples treated with Medical . CONCLUSION: A single use of Medical Tabs for Dentures is effective in eliminating certain species of microorganisms, including selected viruses, in vitro.

J Pept Sci, 2004 Apr, 10(4), 204 - 9
HP(2-9)-magainin 2(1-12), a synthetic hybrid peptide, exerts its antifungal effect on Candida albicans by damaging the plasma membrane; Park Y et al.; In our previous study, HP(2-9)-MA(1-12), HP-MA for short, a hybrid peptide incorporating residues 2-9 of Helicobacter pylori ribosomal protein L1 (HP) and residues 1-12 of magainin 2 (MA) was shown to have strong antibacterial activity . In this study the antifungal activity of HP-MA was evaluated using various fungi, and it was shown that the activity was increased when compared with the parent peptides . In order to investigate the fungicidal mechanism(s) of HP-MA its action against fungal cell membranes was examined by the potassium-release test, which showed that HP-MA caused an increase in the amount of K+ released from the cells . Furthermore, HP-MA induced significant morphological changes . These facts suggested that the fungicidal effect of HP-MA involves damaging the fungal cell membranes . CD investigators suggested that the alpha-helical structure of these peptides plays an important role in their antibiotic effect, but that alpha-helicity is less directly correlated with the enhanced antibiotic activity of the hybrid.

Nippon Ishinkin Gakkai Zasshi, 2004, 45(2), 113 - 9
Inductions of germ tube and hyphal formations are controlled by mRNA synthesis inhibitor in Candida albicans; Imanishi Y et al.; Candida albicans is a pathogenic dimorphic fungus . When yeast cells were pre-incubated in YPD medium at 25degreesC and released into HFM7 medium containing 4% serum at 37degreesC, germ tubes emerged within 0.5 h . To determine whether mRNA or protein synthesis was necessary for germ tube formation, we examined the effects of mRNA and protein syntheses inhibitors on this formation . In the presence of cycloheximide, cells were unbudded and no germ tube was observed . However, in the presence of actinomycin D, germ tube formation was observed while budding growth and true hyphae elongation were blocked . Next, we measured mRNA or protein accumulation during induction of germ tube formation in the presence of the inhibitors . In the presence of cycloheximide, protein was not synthesized, while in the presence of actinomycin D, mRNA synthesis decreased to 6.3% and protein synthesis to 37.7% . The condition we found which allows only germination but not budding or filamentation might be convenient to use in screening genes involved in the initial stage of morphological change in C . albicans.

Nippon Ishinkin Gakkai Zasshi, 2004, 45(2), 55 - 8
{Molecular taxonomy and identification of pathogenic fungi based on DNA sequence analysis}; Sugita T et al.; Although approximately 80,000 fungi are known, less than 1% are associated with human infection . However, their taxonomy has long been insufficient . During the last decade, DNA sequence analysis was introduced to the taxonomy of pathogenic fungi . Taxonomic advances in the field of medical mycology are helping to identify the causative agents of infectious diseases accurately, facilitating diagnosis and treatment . For example, Malassezia furfur was long considered the major microflora in atopic dermatitis, yet recent studies have indicated that this is not the case, as M . furfur is taxonomically heterogeneous and consists of five species . DNA sequence analysis resolved its taxonomic heterogeneity . Similar examples can be seen in "Candida albicans and C . dubliniensis" and "Trichosporon cutaneum and T . asahii" . DNA sequence analysis also enables accurate identification of fungi . At present, almost all pathogenic fungi can be identified by determining the D1/D2 26S rDNA and ITS region of rRNA gene . This paper describes the practical taxonomy and identification of pathogenic fungi based on DNA sequence analysis.

Yeast, 2004 Apr 30, 21(6), 473 - 82
Functional analysis of the Candida albicans ALS1 gene product; Loza L et al.; ALS1 encodes a cell surface protein that mediates adherence of Candida albicans to endothelial cells . The predicted Als1p has an N-terminal region, which contains a signal peptide; a middle region, which contains 20 36-amino acid tandem repeats; and a C-terminal region, which contains a glycosylphosphotidylinositol-anchorage sequence . We used site-directed mutagenesis to delineate the regions in Als1p required for endothelial cell adherence and cell surface expression of the protein . Mutant alleles of ALS1 containing either deletions or insertions were expressed in the normally non-adherent Saccharomyces cerevisiae . These transformants were analysed for endothelial cell adherence and cell surface expression of Als1p . We found that mutations centred around amino acid 285 in the N-terminus completely abolished adherence, but had no effect on cell surface expression of Als1p . Deletion of 15 of the tandem repeats reduced adherence by 50%, whereas deletion of all abolished adherence completely, even though cell surface expression of the N-terminus of Als1p was maintained . Insertions into the C-terminus at amino acids 413 and 254 upstream of the stop codon resulted in a modest loss of adherence, while cell surface expression of Als1p was maintained . An insertion at amino acid 249 in the C-terminus caused complete loss of both adherence and cell surface expression, even though the glycosylphosphotidylinositol-anchorage sequence remained intact . These data suggest a model of Als1p in which the endothelial cell binding region is localized within its N-terminus, the tandem repeats are essential for the proper presentation of the binding site, and the C-terminus is required for localizing Als1p to the cell surface .

Yeast, 2004 Apr 15, 21(5), 429 - 36
Cassettes for the PCR-mediated construction of regulatable alleles in Candida albicans; Gerami-Nejad M et al.; The recent availability of genome sequence information for the opportunistic pathogen Candida albicans has greatly facilitated the ability to perform genetic manipulations in this organism . Two important molecular tools for studying gene function are regulatable promoters for generating conditional mutants and fluorescent proteins for determining the subcellular localization of fusion gene products . We describe a set of plasmids containing promoter-GFP cassettes (P(MET3)-GFP, P(GAL1)-GFP, and P(PCK1)-GFP), linked to a selectable nutritional marker gene (URA3) . PCR-mediated gene modification generates gene-specific promoter, or gene-specific promoter-GFP, fusions at the 5'-end of the gene of interest . One set of primers can be used to generate three strains expressing a native protein of interest, or an amino-terminal GFP-tagged version, from three different regulatable promoters . Thus, these promoter cassette plasmids facilitate construction of conditional mutant strains, overexpression alleles and/or inducible amino-terminal GFP fusion proteins .

Mol Genet Genomics, 2004 Jun, 271(5), 554 - 65 Epub 2004 Apr 28.
A proteomic approach to understanding the development of multidrug-resistant Candida albicans strains; Kusch H et al.; Resistance of the pathogenic yeast Candida albicans to the antifungal agent fluconazole is often caused by the overexpression of genes that encode multidrug efflux pumps ( CDR1, CDR2, or MDR1) . We have undertaken a proteomic approach to gain further insight into the regulatory network controlling efflux pump expression and drug resistance in C . albicans . Three pairs of matched fluconazole-susceptible and resistant clinical C . albicans isolates, in which drug resistance correlated with stable activation of MDR1 or CDR1/2, were analyzed for differences in their protein expression profiles . In two independent, MDR1-overexpressing, strains, additional up-regulated proteins were identified, which are encoded by the YPR127 gene and several members of the IFD ( YPL088) gene family . All are putative aldo-keto reductases of unknown function . These proteins were not up-regulated in a fluconazole-resistant strain that overexpressed CDR1 and CDR2 but not MDR1, indicating that expression of the various efflux pumps of C . albicans is controlled by different regulatory networks . To investigate the possible role of YPR127 in the resistance phenotype of the clinical isolates, we constitutively overexpressed the gene in a C . albicans laboratory strain . In addition, the gene was deleted in a C . albicans laboratory strain and in one of the drug-resistant clinical isolates in which it was overexpressed . Neither forced overexpression nor deletion of YPR127 affected the susceptibility of the strains to drugs and other toxic substances, suggesting that the regulatory networks which control the expression of efflux pumps in C . albicans also control genes involved in cellular functions not related to drug resistance.

PDA J Pharm Sci Technol, 2004 Mar-Apr, 58(2), 75 - 80
Validation of microbial recovery from hydrogen peroxide-sterilized air; Ohresser S et al.; The use of hydrogen peroxide as a sanitant in isolators and other barrier systems is well documented . To confirm that the isolator maintains a germ-free environment between decontamination cycles, microbiological air monitoring is performed after the sanitation and aeration cycles . In this study, we have shown that residual levels of hydrogen peroxide as low as 1 ppm can remain in the isolator and inhibit the growth of microorganisms after concentration on agar media . This lingering hydrogen peroxide can make accurate microbiological air monitoring difficult and can even cause false negative test results . To solve this issue, we have developed a new media that can mediate the effects of residual peroxide and prevent false negative test results . Initially, catalase was tested as a neutralizing agent but proved not to be efficient enough . Instead, 1% pyruvate was added, which was able to tolerate as much as 15 ppm Vaporous Hydrogen Peroxide (VHP) and ensured growth promotion of Staphylococcus aureus, Pseudomonas aeruginosa, Micrococcus luteus, Bacillus subtilis, Candida albicans and Aspergillus niger . The 1% pyruvate retained its neutralizing activity for Micrococcus luteus at up to 100 ppm VHP . Raising the pyruvate concentration to 5% pyruvate enabled neutralization of up to 300 ppm VHP, permitting subsequent growth of Micrococcus luteus on agar media.

J Antibiot (Tokyo), 2004 Feb, 57(2), 117 - 24
New beauvericins, potentiators of antifungal miconazole activity, Produced by Beauveria sp . FKI-1366 . II . Structure elucidation; Fukuda T et al.; The structures of beauvericins D, E and F, novel potentiators of miconazole activity against Candida albicans produced by Beauveria sp . FKI 1366, were elucidated by various spectroscopic analyses including UV, NMR, and MS and degradation experiments . They have the common skeleton of the 18-membered cyclodepsipeptides.

J Antibiot (Tokyo), 2004 Feb, 57(2), 110 - 6
New beauvericins, potentiators of antifungal miconazole activity, Produced by Beauveria sp . FKI-1366 . I . Taxonomy, fermentation, isolation and biological properties; Fukuda T et al.; Three new beauvericins, designated beauvericins D, E and F, were isolated along with known beauvericin and beauvericin A, from the culture of Beauveria sp . FKI-1366 by solvent extraction, ODS column chromatography and HPLC . These compounds potentiate miconazole activity against not only wild Candida albicans but also fluconazole resistant C . albicans . Beauvericins D and E decreased the IC50 value of miconazole against fluconazole resistant C . albicans from 1.3 microM to 0.25 and 0.31 microM, respectively.

Minerva Chir, 2004 Feb, 59(1), 45 - 51
{Postoperative pneumonia caused by Candida in 500 lung surgery patients}; Rovera F et al.; BACKGROUND: Postoperative infections continue to be one of the most frequent complications in hospitalized patients . The incidence of fungal infection has been steadily rising . While Candida albicans remains the most common yeast species isolated in hospitalized patients, other Candida species have been increasingly isolated . METHODS: From 1996 to 2001, a prospective study of 500 consecutive lung surgery patients treated by the same surgical team was conducted to monitor the number of postoperative infections and to evaluate the epidemiology of bacterial and fungal infections . RESULTS: At least one postoperative infection developed in 18% of patients . Of the 51 patients who developed postoperative pneumonia, 24 underwent microbiological examination of sputum . In 19 of them, the culture yielded isolates of one or more Candida species . Slightly under half of patients (47%) with cultures positive for mycetes received treatment with fluconazole until the clinical infection resolved . The drug was well tolerated by all treated patients . CONCLUSIONS: The incidence of fungal infection has increased also at our center . In 19 of 24 patients with postoperative pneumonia, sputum cultures yielded Candida species isolates . Many factors may be contributing to the rise in fungal infections after surgery . As pneumonia caused by Candida led to a significantly longer length of hospital stay in our case series, we draw attention to the importance of early diagnosis of postoperative mycotic lung disease in order to institute timely and targeted therapy.

Eur J Med Chem, 2004 May, 39(5), 453 - 8
Synthesis and biological evaluation of oxindoles and benzimidazolinones derivatives; Messaoudi S et al.; The synthesis of new oxindoles and benzimidazolinones derivatives bearing a sugar residue on the aromatic nitrogen is described . The presence of the glycoside moiety should enhance the solubility of these heterocyclic compounds and/or improve the interaction with the active site of the biological targets . The inhibitory activities of these new compounds toward five kinases were examined: KDR (VEGFR-2), FGFR-1, PDGFR-beta, EGFR and Tie 2 . Furthermore, the antibacterial activities of the prepared compounds were tested against two Gram-positive bacteria Bacillus cereus and Streptomyces chartreusis, a Gram-negative bacterium Escherichia coli and a yeast Candida albicans.

Biochim Biophys Acta, 2004 May 3, 1672(2), 76 - 85
Proteolytic fragments of ovalbumin display antimicrobial activity; Pellegrini A et al.; Ovalbumin, one of the major proteins present in avian egg white, was proteolytically digested by trypsin and chymotrypsin and the peptide fragments were investigated for their antimicrobial activity . The antimicrobial peptides were isolated and characterized . From the tryptic digestion, the following five antimicrobial peptide fragments were obtained: SALAM (residues 36-40), SALAMVY (residues 36-42) YPILPEYLQ (residues 111-119), ELINSW (residues 143-148) and NVLQPSS (residues 159-165) . Digestion of ovalbumin by chymotrypsin yielded the antimicrobial peptides AEERYPILPEYL (residues 127-138), GIIRN (residues 155-159) and TSSNVMEER (residues 268-276) . The peptides were synthesized and found to exert antimicrobial activity . They were strongly active against Bacillus subtilis and to a lesser extent against the other bacterial strains examined . A weak fungicidal activity against Candida albicans was also shown by some peptides . Ovalbumin itself was not bactericidal against all the bacteria strains examined . Our results suggest that the food protein ovalbumin may supply the organism with antimicrobial peptides, supporting the immunodefences of the organism.

Biochem J, 2004 Jul 15, 381(Pt 2), 447 - 52
Reactive oxygen species play no role in the candidacidal activity of the salivary antimicrobial peptide histatin 5; Veerman EC et al.; The mechanism of action of antimicrobial peptides is still a matter of debate . The formation of ROS (reactive oxygen species) has been suggested to be the crucial step in the fungicidal mechanism of a number of antimicrobial peptides, including histatin 5 and lactoferrin-derived peptides . In the present study we have investigated the effects of histatin 5 and of a more amphipathic synthetic derivative, dhvar4, on the generation of ROS in the yeast Candida albicans, using dihydroethidium as an indicator for ROS . With both peptides, a substantial enhancement of fluorescence was observed . However, TEMPO (2,2,6,6-tetramethylpiperidine-N-oxyl), a cell-permeant ROS scavenger, did not have an inhibitory effect on killing or on the enhancement of fluorescence . Furthermore, antimycin and azide, which have been reported to induce ROS in vitro, were not able to enhance the dihydroethidium fluorescence, while chlorhexidine, a non-specific antiseptic agent, enhanced dihydroethidium fluorescence to the same extent as did the peptides . Fluorescence microscopy showed the fluorescence enhancement to be a consequence of the release of unbound preformed ethidium from the mitochondrial matrix within the cell . It is concluded that ROS do not play a role in the histatin 5-mediated killing of C . albicans.

Surg Today, 2004, 34(5), 466 - 9
Candida albicans-infected pancreatic pseudocyst: report of a case; Zulfikaroglu B et al.; We report the case of a 48-year-old man with a pseudocyst infected by Candida albicans, and review the relevant literature . The patient was successfully treated by a Roux-en-Y cystojejunostomy of the pseudocyst and adjunctive therapy with amphotericin B . Candida species isolated from a pancreatic pseudocyst or abscess should be considered pathogenic, and treated aggressively.

Antimicrob Agents Chemother, 2004 May, 48(5), 1778 - 87
Membrane sphingolipid-ergosterol interactions are important determinants of multidrug resistance in Candida albicans; Mukhopadhyay K et al.; In this study, we examined the importance of membrane ergosterol and sphingolipids in the drug susceptibilities of Candida albicans . We used three independent methods to test the drug susceptibilities of erg mutant cells, which were defective in ergosterol biosynthesis . While spot and filter disk assays revealed that erg2 and erg16 mutant cells of C . albicans became hypersensitive to almost all of the drugs tested (i.e., 4-nitroquinoline oxide, terbinafine, o-phenanthroline, itraconazole, and ketoconazole), determination of the MIC at which 80% of the cells were inhibited revealed more than fourfold increase in susceptibility to ketoconazole and terbinafine . Treatment of wild-type C . albicans cells with fumonisin B1 resulted in 45% inhibition of sphingolipid biosynthesis and caused cells to become hypersensitive to the above drugs . Although erg mutants displayed enhanced membrane fluidity and passive diffusion, these changes alone were not sufficient to elicit the observed hypersusceptibility phenotype of erg mutants . For example, the induction in vitro of a 12% change in the membrane fluidity of C . albicans cells by a membrane fluidizer, benzyl alcohol, did not affect the drug susceptibilities of Candida cells . Additionally, the surface localization of green fluorescent protein-tagged Cdr1p, a major drug efflux pump protein of C . albicans, revealed that any disruption in ergosterol and sphingolipid interactions also interfered with its proper surface localization and functioning . A 50% reduction in the efflux of the Cdr1p substrate, rhodamine 6G, in erg mutant cells or in cells with a reduced sphingolipid content suggested a strong correlation between these membrane lipid components and this major efflux pump protein . Taken together, the results of our study demonstrate for the first time that there is an interaction between membrane ergosterol and sphingolipids, that a reduction in the content of either of these two components results in a disruption of this interaction, and that this disruption has deleterious effects on the drug susceptibilities of C . albicans cells.

J Nat Prod, 2004 Apr, 67(4), 718 - 20
Antimicrobial chlorinated bibenzyls from the liverwort Riccardia marginata; Baek SH et al.; Three new chlorinated bibenzyls 3, 4, and 5, have been isolated from the New Zealand liverwort Riccardia marginata . This is the first report of simple chlorinated bibenzyls from any natural source . Compounds 3-5 showed antimicrobial activity against Bacillus subtilis, Candida albicans, and Trichophyton mentagrophytes.

Infect Immun, 2004 May, 72(5), 2703 - 9
Dialysis unmasks the fungicidal properties of glandular salivary secretions; Helmerhorst EJ et al.; Several salivary proteins exhibit fungicidal activity against the opportunistic oral pathogen Candida albicans when they are tested as pure proteins in vitro . However, salivary secretions that are examined by the same assays either lack or exhibit very low candidacidal activity . Since ionic strength is known to have an inhibitory effect on the fungicidal activities of some proteins, parotid secretion was subjected to dialysis with membranes having molecular weight cutoffs (MWCOs) of 500, 1000, 10000, and 25000 . Dialysis with membranes with MWCOs of >=1000 promoted fungicidal activity of parotid secretion, and this activity was dose dependent . The addition of sodium chloride to dialyzed, fungicidal parotid secretion abolished this activity, indicating that the fungicidal component was salt sensitive . Similar results were obtained with submandibular and sublingual secretions . Polyacrylamide gel electrophoresis under native and denaturing conditions was used to analyze the composition of the dialysate . Unexpectedly, proteins with MWs much lower than the nominal MWCOs of the membranes were not lost during dialysis . Among the retained proteins, the two fractions with MWs of approximately 17000 and 4000 exhibited fungicidal activity . These results are consistent with the presence of lysozyme and histatins, respectively, which may represent the major candidacidal capacity of dialyzed parotid secretion.

Infect Immun, 2004 May, 72(5), 2513 - 20
Inhibition of monocytic interleukin-12 production by Candida albicans via selective activation of ERK mitogen-activated protein kinase; Tang N et al.; Our previous data demonstrated that live Candida albicans inhibits interleukin-12 (IL-12) production by human monocytes . Here we explored whether C . albicans inhibits IL-12 via a released factor and whether the inhibition is mediated via mitogen-activated protein kinase (MAPK) regulation . Supernatant fluids were obtained from cultured C . albicans (SC5314) as well as cultured Saccharomyces cerevisiae after 20 h of incubation . At 2 h postincubation of monocytes with heat-killed C . albicans (HKCA) (2:1) to stimulate IL-12, concentrated fungal supernatant fluids were added and incubated for an additional 20 h . The present data show that, unlike supernatant fluids obtained from S . cerevisiae, the C . albicans supernatant fluids significantly suppressed IL-12 production induced by HKCA . This suggested that the inhibition is Candida specific . A preliminary biochemical analysis revealed that this secretory IL-12 inhibitory factor is glycoprotein in nature . The inhibitory activity had no effect on the phagocytosis of yeasts . Supernatant fluids from C . albicans markedly induced the phosphorylation of ERK44/42 MAPK, but not p38 and SAPK, 1 min after they were added to monocytes . To test if the induction of ERK44/42 MAPK was central to the IL-12 inhibition, we used gamma interferon (IFN-gamma) (1 ng/ml) plus lipopolysaccharide (LPS) (100 ng/ml) to stimulate IL-12 production by monocytes . The inhibition of ERK MAPK by the specific inhibitor PD 98059 significantly reduced phospho-ERK44/42 MAPK levels induced by C . albicans supernatant fluids in the IFN-gamma-plus-LPS-driven monocytes . Concomitantly, PD 98059 reversed the IL-12 inhibitory activity of the C . albicans supernatant (P < 0.01) . These data indicate that C . albicans can inhibit IL-12 production by secreting an ERK44/42 MAPK-stimulating factor and thus can attenuate effective immune responses.

J Immunol, 2004 May 1, 172(9), 5622 - 8
Immune-deficient Drosophila melanogaster: a model for the innate immune response to human fungal pathogens; Alarco AM et al.; We explored the host-pathogen interactions of the human opportunistic fungus Candida albicans using Drosophila melanogaster . We established that a Drosophila strain devoid of functional Toll receptor is highly susceptible to the human pathogen C . albicans . Using this sensitive strain, we have been able to show that a set of specific C . albicans mutants of different virulence in mammalian infection models are also impaired in virulence in Drosophila and remarkably display the same rank order of virulence . This immunodeficient insect model also revealed virulence properties undetected in an immunocompetent murine model of infection . The genetic systems available in both host and pathogen will enable the identification of host-specific components and C . albicans genes involved in the host-fungal interplay.

J Ethnopharmacol, 2004 May, 92(1), 107 - 11
Composition and bioactivity of the leaf essential oil of Heteropyxis dehniae from Zimbabwe; Sibanda S et al.; The leaf oil of Heteropyxis dehniae Suess . (Heteropyxidaceae) was obtained by hydrodistillation and analyzed by GC/MS . The most abundant essential oil components are linalool (58.3%), 4-terpineol (9.8%), alpha-terpineol (3.6%), and caryophyllene oxide (3.1%) . The antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, and Aspergillus niger, and the in vitro cytotoxicity of the oil on PC-3, MDA-MB-231, Hs 578T, MCF7, SK-MEL-28, and 5637 human tumor cells were also examined . Caryophyllene oxide shows notable cytotoxic activity with LC50 values of 147-351 microM .

J Ethnopharmacol, 2004 May, 92(1), 53 - 6
Evaluation of the bioactivity of triterpene mixture isolated from Carmona retusa (Vahl.) Masam leaves; Villasenor IM et al.; The major constituent of Carmona retusa (Vahl.) Masam . leaves is an intractable mixture of triterpenes, namely alpha-amyrin (43.7%), beta-amyrin (24.9%), and baurenol (31.4%) . At a dosage of 100mg/kg mouse, the triterpene mixture exhibited 51% analgesic activity but only showed 20% anti-inflammatory activity . Kruskal-Wallis one-way analysis of variance by ranks showed that the triterpene mixture is as active as mefenamic acid, a commercially available analgesic, at alpha = 0.01 . The charcoal tracing test showed a 29% anti-diarrheal activity for the triterpene mixture, which increased to 55% at a dosage of 250 mg/kg mouse . At the higher dosage, the triterpene mixture differed significantly from its solvent control at alpha = 0.01 . Results of the micronucleus test showed that the triterpene mixture did not exhibit mutagenic nor anti-mutagenic activity at alpha = 0.001 . There was no significant decrease in blood glucose levels (bgl) in alloxan-induced diabetic mice after administration of the triterpene mixture . The triterpene mixture was inactive against Escherichia coli and possessed moderate activities against Staphylococcus aureus, Candida albicans, and Trichophyton mentagrophytes .

J Eur Acad Dermatol Venereol, 2004 May, 18(3), 301 - 4
Superficial mycoses and HIV infection in Yaounde; Lohoue Petmy J et al.; The incidence of superficial infections in patients with human immunodeficiency virus (HIV) is increasing in Yaounde . To determine the prevalence of superficial mycoses in HIV-positive subjects, a study was carried out in different hospital settings in Yaounde over a period of 18 months . Clinical observations were followed by the culture of samples on Sabouraud's glucose agar . Of the 148 HIVab-positive patients examined, 79 (44 males and 35 females) had at least one superficial mycosis (53%) . The clinical presentations and the causative organisms did not differ from those found in immunocompetent individuals . The main lesions were oral candidiasis (77%), tinea corporis (21%), tinea versicolor (15%), tinea pedis (13%) and tinea inguium (12%) . Trichophyton rubrum and Candida albicans were the organisms most commonly isolated . These results indicate the benefit of studying superficial mycoses for optimal management of HIV patients.

FEMS Immunol Med Microbiol, 2004 May 1, 41(1), 27 - 34
Anti-retroviral therapy with protease inhibitors decreases virulence enzyme expression in vivo by Candida albicans without selection of avirulent fungus strains or decreasing their anti-mycotic susceptibility; De Bernardis F et al.; Highly active anti-retroviral therapies (HAART) with human immunodeficiency virus (HIV) protease inhibitors (PIs) or nonnucleoside reverse-transcriptase inhibitors (NNRTI) were compared for their effect on prevalence, aspartyl proteinase (Sap) production and the biotypes and anti-mycotic sequential susceptibility of Candida spp . isolates from the oral cavity in a longitudinal prospective study . HAART-PI, but not HAART-NNRTI strongly inhibited Sap expression in the oral cavity without exerting any consistent effect on the role of Candida spp . isolation or selection of low virulence or anti-mycotic resistant fungus biotype . More importantly, the sequential isolates of Candida albicans from HAART-PI, but not those from suspended HAART-NNRTI, showed an increased Sap production in vitro . While further demonstrating that HIV-PI inhibit Sap expressions, our results do not support the view that the mentioned inhibition could eliminate Candida or its selection of the oral cavity.

Vet Ophthalmol, 2004 May-Jun, 7(3), 159 - 62
Mycotic endophthalmitis in a dog caused by Candida albicans; Linek J; A case of mycotic endophthalmitis in the dog caused by Candida albicans is presented . The 3-year-old dog had a history of bloody diarrhea 3 months previously . The dog presented with acute signs of unilateral panuveitis . Aqueocentesis, vitreocentesis, and routine blood tests were performed but did not contribute to the diagnosis . The posterior segment could not be visualized because of flare and fibrin . On day 7 ultrasonography showed retinal separation which progressed to vitreous compartmentalization and abscessation by day 14 . Three weeks after onset, glaucoma developed and enucleation was performed . Histology revealed the yeast Candida to be the causative agent . Post-enucleation serum Candida antibody titer was 1 : 640 (human threshold 1 : 120), as determined by agglutination test . A relapse of enteric signs 3 months later led to the diagnosis of chronic lymphocytic enteritis . An hematogenous route of infection is suspected.

Biometals, 2004 Apr, 17(2), 115 - 20
Antimicrobial activities of N-(2-hydroxy-1-naphthalidene)-amino acid(glycine, alanine, phenylalanine, histidine, tryptophane) Schiff bases and their manganese(III) complexes; Sakiyan I et al.; The in vitro antibacterial and antifungal activities of five different amino acid Schiff bases derived from the reaction of 2-hydroxy-1-naphthaldehyde with glycine, L-alanine L-phenylalanine, L-histidine, L-tryptophane and the manganese(III) complexes of these bases were investigated . Structures of the Schiff bases were proven by 1H-NMR . In vitro activities against some Gram-positive (Staphylococcus aureus and Bacillus polymyxa) and Gram-negative (Escherichia coli) bacteria and the fungus Candida albicans were determined . The antimicrobial activities tended to decrease with the increasing size of the amino acid residues.

Allergol Immunopathol (Madr), 2004 Mar-Apr, 32(2), 82 - 5
{Otomastoiditis candidiasica and hyper IgE syndrome}; Vasquez C et al.; PATIENT: A 5-year-old girl presented with chronic otorrhea, cervical adenopathies and cellulitis of the knee.In addition to these lesions, physical examination revealed eczema on the scalp, neck, perineal and umbilical regions and the persistence of deciduous teeth with adult teeth (double dental arch).Complementary investigations showed the following concentrations: IgE 23969 UI/l, IgD 440 U/L, IgG 23000 mg/L, and IgA 4220 mg/L . Intradermal skin testing to Candida was negative and the results of the remaining immunological studies were normal . Computerized axial tomography revealed bilateral otomastoiditis . Candida albicans was isolated from ear secretion cultures . The definitive diagnosis was hyper IgE syndrome . The patient responded favorably to antibiotic and antifungal therapy and is currently undergoing period outpatient monitoring.

Ann Pharmacother, 2004 Jun, 38(6), 973 - 7 Epub 2004 Apr 14.
Pediatric case series evaluating a standardized Candida albicans skin test product; Ohri LK et al.; BACKGROUND: A ready-to-use, standardized Candida albicans skin test product (Candin) first became available in 1995 . As of April 6, 2004, no published reports have been found describing efficacy or safety with pediatric use of this reagent . OBJECTIVE: To present a case series describing experience with use of Candin to assess 78 pediatric cases for delayed-type hypersensitivity . METHODS: The standardized C . albicans reagent was one of up to 4 antigens used to evaluate patients' cellular immunity . Candin was used with purified protein derivative tuberculosis testing for 76 patients (4 mo-16 y of age) and for anergy testing alone in 2 cases . Candin was used with at least one other skin test reagent for 24 subjects . RESULTS: Fifty-three of 78 subjects (68%) responded to at least one skin test antigen . Candin had an overall response rate of 64% (50/78) . There was a response to Candin in 54% (13/24) of subjects with multiple reagents applied, the highest rate of any antigen used . There was a 27% (3/11) positive Candin response for subjects <1 year of age; this compared with 21% and 23% in 2 published reports on use of nonstandardized tests for this age group . No adverse events were associated with use of Candin . CONCLUSIONS: Candin testing gives similar or better response rates compared with published data on older, nonstandardized C . albicans reagents and other anergy test antigens used in this case series . It should be used in combination with multiple other control antigens to most effectively assess for anergy, particularly in infants, as well as in patients recognized to be immune deficient.

Clin Microbiol Rev, 2004 Apr, 17(2), 281 - 310
Surface glycans of Candida albicans and other pathogenic fungi: physiological roles, clinical uses, and experimental challenges; Masuoka J; Although fungi have always been with us as commensals and pathogens, fungal infections have been increasing in frequency over the past few decades . There is a growing body of literature describing the involvement of carbohydrate groups in various aspects of fungal disease . Carbohydrates comprising the cell wall or capsule, or as a component of glycoproteins, are the fungal cell surface entities most likely to be exposed to the surrounding environment . Thus, the fungus-host interaction is likely to involve carbohydrates before DNA, RNA, or even protein . The interaction between fungal and host cells is also complex, and early studies using whole cells or crude cell fractions often produced seemingly conflicting results . What was needed, and what has been developing, is the ability to identify specific glycan structures and determine how they interact with immune system components . Carbohydrate analysis is complicated by the complexity of glycan structures and by the challenges of separating and detecting carbohydrates experimentally . Advances in carbohydrate chemistry have enabled us to move from the foundation of composition analysis to more rapid characterization of specific structures . This, in turn, will lead to a greater understanding of how fungi coexist with their hosts as commensals or exist in conflict as pathogens.

Mol Cell Biol, 2004 May, 24(9), 4075 - 82
Targeted disruption of SPI3/Serpinb6 does not result in developmental or growth defects, leukocyte dysfunction, or susceptibility to stroke; Scarff KL et al.; Protease inhibitor 6 (PI-6/SERPINB6) is a widely expressed nucleocytoplasmic serpin . It inhibits granulocyte cathepsin G and neuronal neuropsin, and it is thought to protect cells from death caused by ectopic release or internalization of protease during stress such as infection or cerebral ischemia . To probe the biological functions of PI-6, we generated mice lacking its ortholog (SPI3/Serpinb6) . SPI3-deficient mice developed normally and were fertile, and no abnormal pathology or increased sensitivity to cerebral ischemia was observed . There were no perturbations in leukocyte development or numbers, and recruitment of leukocytes to the peritoneal cavity was normal . SPI3-deficient mice were equally susceptible as wild-type mice to systemic Candida albicans infection, although there was a slight decrease in the ability of neutrophils from SPI3-deficient mice to kill C . albicans in vitro . Increased levels of a related inhibitor Serpinb1 (monocyte/neutrophil elastase inhibitor) in the tissues of targeted mice suggests that compensation by other serpins reduces the impact of SPI3 deficiency in these animals and may explain the lack of a more obvious phenotype.

Mycoses, 2004 Apr, 47(3-4), 131 - 5
Onychomycosis in Denmark: prevalence of fungal nail infection in general practice; Svejgaard EL et al.; The study was initiated in order to get knowledge of the frequency of onychomycosis in patients visiting general practitioners in Denmark . A study design was using a display showing photos of abnormal nails including fungal infection, a clinical examination and a questionnaire . The practitioners obtained nail material . Direct microscopy and culture as well as histopathology, were carried out blindly in two different mycological laboratories . A number of 8546 patients were seen during the 6 months of the study, 5755 (67.3%) took part in the investigation . Clinical abnormal nails were observed in 948 (16.5%) patients, 52% males and 48% females, aged 18-92, mean 55 years old . Onychomycosis caused by dermatophytes were found in 238 (4.1%) and by Candida albicans in 45 (0.8%) . Trichosporon cutaneum and Scopulariopsis brevicaulis were isolated each in 15 cases as single cultures . Onychomycosis, was typically seen in toenails as the distal-lateral type in males more than 40 years old . Predisposing factors were familiar dermatophytosis (22%), trauma (16.9%), diabetes mellitus (6.7%) and peripheral circulatory insufficiency (5.9%).

Mycoses, 2004 Apr, 47(3-4), 115 - 20
Effect of thyroliberin on the course of experimental candidosis in mice; Blaszkowska J et al.; Thyroliberin (TRH) is one of the hormones, which affect immunologic processes . This hormone was studied in experimental subacute candidosis in mice . BALB/c males were given intraperitoneally single dose of Candida albicans cells (1 x 10(7) to 1 x 10(9) cfu of strain no . 244-33 ATCC) . The animals from the experimental groups were injected subcutaneously, after 24 h from inoculation, dose of 10 microg TRH in 0.2 ml 0.9% NaCl, seven times at 24 h intervals . The control animals were given respectively 0.2 ml of physiologic NaCl solution . We have found that the examined hormone significantly decreases mortality in these animals (LD50 C . albicans for mice treated with TRH was three times higher than that in the control groups), prolongs mean survival time for mice and decreases the intensity of fungal invasion of the animal organs.

J Chemother, 2004 Feb, 16(1), 38 - 44
Tetracycline in combination with sodium dioctylsulfosuccinate show increased antimicrobial activity in resistant microorganisms; Simonetti G et al.; We present evidence that sodium dioctylsulfosuccinate, at non inhibitory concentrations (1000 mg/L), is able to increase the antimicrobial activity of tetracycline in non susceptible bacterial and fungal strains . In culture inhibition tests, pretreatment with sodium dioctylsulfosuccinate caused a 52-fold decrease in the geometric mean MIC to tetracycline in 10 Candida albicans strains (p<0.01), a 165-fold decrease in the geometric mean MIC to tetracycline in 10 E . coli strains (p<0.001) and a significant decrease in the mean MIC of 3 strains of Candida krusei and Candida glabrata . In microbicidal tests, tetracycline in association with sodium dioctylsulfosuccinate killed 10(4) cfu tetracycline-resistant Candida albicans in 15 min and 10(4) CFU resistant E . coli in 3 min (p<0.001) . Furthermore, in the N-acetyl-D-glucosamine test to calculate the number of hyphal cells, a combination of tetracycline (50 mg/L) (non inhibitory concentrations) plus sodium dioctylsulfosuccinate (25 mg/L) caused a 50-fold increase in the inhibition of hyphal cells in C . albicans (p<0.001); C . albicans cells treated with tetracycline plus sodium dioctylsolfosuccinate annulled the cell surface hydrophobicity (p<0.001) . This increase in antimicrobial activity may be attributed to impairment and alteration of the membrane barrier within the microorganisms and a depletion of the thiol groups (p<0.001) critical to their efficiency.

Eukaryot Cell, 2004 Apr, 3(2), 471 - 82
Polarized hyphal growth in Candida albicans requires the Wiskott-Aldrich Syndrome protein homolog Wal1p; Walther A et al.; The yeast-to-hypha transition is a key feature in the cell biology of the dimorphic human fungal pathogen Candida albicans . Reorganization of the actin cytoskeleton is required for this dimorphic switch in Candida . We show that C . albicans WAL1 mutants with both copies of the Wiskott-Aldrich syndrome protein (WASP) homolog deleted do not form hyphae under all inducing conditions tested . Growth of the wild-type and wal1 mutant strains was monitored by in vivo time-lapse microscopy both during yeast-like growth and under hypha-inducing conditions . Isotropic bud growth produced round wal1 cells and unusual mother cell growth . Defects in the organization of the actin cytoskeleton resulted in the random localization of actin patches . Furthermore, wal1 cells exhibited defects in the endocytosis of the lipophilic dye FM4-64, contained increased numbers of vacuoles compared to the wild type, and showed defects in bud site selection . Under hypha-inducing conditions wal1 cells were able to initiate polarized morphogenesis, which, however, resulted in the formation of pseudohyphal cells . Green fluorescent protein (GFP)-tagged Wal1p showed patch-like localization in emerging daughter cells during the yeast growth phase and at the hyphal tips under hypha-inducing conditions . Wal1p-GFP localization largely overlapped with that of actin . Our results demonstrate that Wal1p is required for the organization of the actin cytoskeleton and hyphal morphogenesis in C . albicans as well as for endocytosis and vacuole morphology.

J Infect Dis, 2004 Apr 15, 189(8), 1524 - 7 Epub 2004 Apr 01.
Recombinant interleukin-18 protects against disseminated Candida albicans infection in mice; Stuyt RJ et al.; Endogenous interleukin (IL)-18 is necessary for host defense against candidiasis . Prophylactic treatment of Candida albicans-infected mice with recombinant murine (r) IL-18 decreased mortality, which was accompanied by a decreased outgrowth of yeasts in the kidneys 1 day after infection . Therapeutic administration of rIL-18 also resulted in decreased outgrowth of C . albicans in the kidneys and increased levels of interferon- gamma, both in the circulation and after in vitro stimulation of splenocytes with C . albicans . Histopathologic analysis of the kidneys showed increased inflammation and decreased growth of C . albicans in rIL-18-treated mice . In conclusion, rIL-18 improves outcome of disseminated candidiasis in mice and may prove useful as adjuvant immunotherapy of fungal infections.

Microbiology, 2004 Apr, 150(Pt 4), 921 - 8
Independent regulation of chitin synthase and chitinase activity in Candida albicans and Saccharomyces cerevisiae; Selvaggini S et al.; Chitin is an essential structural polysaccharide in fungi that is required for cell shape and morphogenesis . One model for wall synthesis at the growing cell surface suggests that the compliance that is necessary for turgor-driven expansion of the cell wall involves a delicate balance of wall synthesis and lysis . Accordingly, de novo chitin synthesis may involve coordinated regulation of members of the CHS chitin synthase and CHT chitinase gene families . To test this hypothesis, the chitin synthase and chitinase activities of cell-free extracts were measured, as well as the chitin content of cell walls isolated from isogenic mutant strains that contained single or multiple knock-outs in members of these two gene families, in both Candida albicans and Saccharomyces cerevisiae . However, deletion of chitinase genes did not markedly affect specific chitin synthase activity, and deletion of single CHS genes had little effect on in vitro specific chitinase activity in either fungus . Chitin synthesis and chitinase production was, however, regulated in C . albicans during yeast-hypha morphogenesis . In C . albicans, the total specific activities of both chitin synthase and chitinase were higher in the hyphal form, which was attributable mainly to the activities of Chs2 and Cht3, respectively . It appeared, therefore, that chitin synthesis and hydrolysis were not coupled, but that both were regulated during yeast-hypha morphogenesis in C . albicans.

EMBO J, 2004 Apr 21, 23(8), 1845 - 56 Epub 2004 Apr 08.
Hgc1, a novel hypha-specific G1 cyclin-related protein regulates Candida albicans hyphal morphogenesis; Zheng X et al.; The human fungal pathogen Candida albicans switches from yeast to hyphal growth when exposed to serum or phagocytosed . However, the importance of this morphological switch for virulence remains highly controversial due to the lack of a mutant that affects hyphal morphogenesis only . Although many genes specifically expressed in hyphal cells have been identified and shown to encode virulence factors, none is required for hyphal morphogenesis . Here we report the first hypha-specific gene identified, HGC1, which is essential for hyphal morphogenesis . Deletion of HGC1 abolished hyphal growth in all laboratory conditions tested and in the kidneys of systemically infected mice with markedly reduced virulence . HGC1 expression is co-regulated with other virulence genes such as HWP1 by the cAMP/protein kinase A signaling pathway and transcriptional repressor Tup1/Nrg1 . Hgc1 is a G1 cyclin-related protein and co-precipitated with the cyclin-dependent kinase (Cdk) CaCdc28 . It has recently emerged that cyclin/Cdk complexes promote other forms of polarized cell growth such as tumor cell migration and neurite outgrowth . C . albicans seems to have adapted a conserved strategy to control specifically hyphal morphogenesis.

J Clin Microbiol, 2004 Apr, 42(4), 1673 - 9
Significance of amplified fragment length polymorphism in identification and epidemiological examination of Candida species colonization in children undergoing allogeneic stem cell transplantation; Ball LM et al.; Candida albicans and non-C . albicans Candida species are increasingly being isolated from patients in high-risk categories, most notably, those who have undergone stem cell transplantation (SCT) . Identification of the presence of non-C . albicans Candida species early in the course of the transplant procedure is important, as these species exhibit different sensitivities to the available antifungal treatments and cause mortality at rates that vary from those for C . albicans . Amplified fragment length polymorphism (AFLP) analysis has been shown to be a reliable method of reproducibly identifying medically important Candida species . We investigated the use of serial AFLP analysis of 54 routine surveillance cultures for the identification and epidemiological examination of Candida sp . colonization in five consecutive children undergoing allogeneic SCT . One child became colonized with a C . albicans strain and remained colonized with this strain during the whole admission period . Another child had persistent colonization with a C . albicans strain with striking variations in its AFLP patterns over time, which was considered indicative of microevolution . Candida dubliniensis, Candida lusitaniae, and Saccharomyces cerevisiae were identified in the three remaining patients, with two children being simultaneously and transiently colonized with different species . These findings show that colonization with yeasts during transplantation is a complex and dynamic interaction between the host and the organism(s) . In our study three strains from eight separate time points were incorrectly identified as C . albicans by a rapid enzyme test . AFLP analysis of surveillance cultures allowed more accurate and informative epidemiological evaluations of pathogenic yeasts in children during transplantation.

J Clin Microbiol, 2004 Apr, 42(4), 1519 - 27
Incidence of bloodstream infections due to Candida species and in vitro susceptibilities of isolates collected from 1998 to 2000 in a population-based active surveillance program; Hajjeh RA et al.; To determine the incidence of Candida bloodstream infections (BSI) and antifungal drug resistance, population-based active laboratory surveillance was conducted from October 1998 through September 2000 in two areas of the United States (Baltimore, Md., and the state of Connecticut; combined population, 4.7 million) . A total of 1,143 cases were detected, for an average adjusted annual incidence of 10 per 100,000 population or 1.5 per 10,000 hospital days . In 28% of patients, Candida BSI developed prior to or on the day of admission; only 36% of patients were in an intensive care unit at the time of diagnosis . No fewer than 78% of patients had a central catheter in place at the time of diagnosis, and 50% had undergone surgery within the previous 3 months . Candida albicans comprised 45% of the isolates, followed by C . glabrata (24%), C . parapsilosis (13%), and C . tropicalis (12%) . Only 1.2% of C . albicans isolates were resistant to fluconazole (MIC, > or = 64 microg/ml), compared to 7% of C . glabrata isolates and 6% of C . tropicalis isolates . Only 0.9% of C . albicans isolates were resistant to itraconazole (MIC, > or = 1 micro g/ml), compared to 19.5% of C . glabrata isolates and 6% of C . tropicalis isolates . Only 4.3% of C . albicans isolates were resistant to flucytosine (MIC, > or = 32 microg/ml), compared to < 1% of C . parapsilosis and C . tropicalis isolates and no C . glabrata isolates . As determined by E-test, the MICs of amphotericin B were > or = 0.38 microg/ml for 10% of Candida isolates, > or =1 microg/ml for 1.7% of isolates, and > or = 2 microg/ml for 0.4% of isolates . Our findings highlight changes in the epidemiology of Candida BSI in the 1990s and provide a basis upon which to conduct further studies of selected high-risk subpopulations.

Phytomedicine, 2004 Feb, 11(2-3), 224 - 9
Trypanocidal and antimicrobial activities of Moquinia kingii; Schinor EC et al.; A chloroform crude extract (aerial part) and two compounds, apigenin (1) and cynaropicrin (2), isolated from Moquinia kingii were evaluated against Trypanosoma cruzi trypomastigotes in vitro . Antimicrobial activity was also screened using twenty-two strains including gram-positive and gram-negative bacteria and the yeasts Candida albicans and C . tropicalis . The chloroform crude extract, fractions and isolated compounds from M . kingii were active for both activities . The IC50 values for trypanocidal activity obtained for cynaropicrin and apigenin were 93.5 microg/ml and 181 microg/ml, respectively, while the minimum inhibitory concentrations (MICs) varied from 100 microg/ml to 2500 microg/ml, against the strains of bacteria and yeasts evaluated.

Stem Cells Dev, 2004 Feb, 13(1), 39 - 50
Candida albicans can stimulate stromal cells resulting in enhanced granulopoiesis; Basu S et al.; Previously, we have reported that although unperturbed granulocyte colony-stimulating factor (GCSF)-deficient (G-CSF-/-) mice are neutropenic, when challenged with Candida albicans, they develop a profound neutrophilia . In an attempt to understand the basis of Candida-induced neutrophilia in G-CSF-deficient mice, we have modified the Dexter bone marrow culture system to produce an in vitro model that mimics emergency granulopoiesis in vivo . In this model, stromal cultures are overlaid with bone marrow cells in the presence or absence of heat-inactivated (HI) Candida . Irrespective of the genotype of mice used as a source of bone marrow-derived stromal cells, stimulation of these cultures with HI Candida led to a significantly greater recovery of cells compared to unstimulated stromal cultures . In addition, there was a marked increase in the number of colony-forming units granulocyte-macrophage (CFU-GM), as well as in the percentage of granulocytes in the population of nonadherent cells recovered from HI Candida-stimulated cultures . The conditioned medium generated from stromal cultures derived from either wild-type or G-CSF-/- mice exposed to HI Candida, when applied to bone marrow cells in a soft agar clonogenic assay stimulated M-, GM-, and G- type colonies . Interleukin-3 (IL-3) and GM-CSF could not be detected in the conditioned medium from either HI Candida stimulated or unstimulated stromal cultures . However, IL-6 was detected in the conditioned media from both wild-type and G-CSF-/- stromal cultures . Addition of anti-IL-6 antibody significantly impaired granulopoiesis in unstimulated and HI Candida-stimulated, wild type, and G-CSF-/- stromal cultures . Conditioned medium generated from G-CSF/IL-6-deficient stromal cells had the capacity to stimulate bone marrow cells to form colonies comprised of granulocytes and macrophages in soft agar clonogenic assay . This study demonstrates that stromal cells can be stimulated with HI Candida and gives an insight into Candida mediated granulopoiesis.

J Immunol, 2004 Apr 15, 172(8), 4934 - 40
Effective targeting of pathogens to neutrophils via chimeric surfactant protein D/anti-CD89 protein; Tacken PJ et al.; Targeting of specific pathogens to FcRs on immune effector cells by using bispecific Abs was reported to result in effective killing of the pathogens, both in vitro and in vivo . Instead of targeting a specific pathogen to an FcR, we assessed whether a broad spectrum of pathogens can be targeted to an FcR using surfactant protein D (SP-D) . SP-D is a collectin that binds a great variety of pathogens via its carbohydrate recognition domain . A recombinant trimeric fragment of SP-D (rfSP-D), consisting of the carbohydrate recognition domain and neck domain of human SP-D, was chemically cross-linked to the Fab' of an Ab directed against the human Fc alpha RI (CD89) . In vitro, the chimeric rfSP-D/anti-CD89 protein enhanced uptake of Escherichia coli, Candida albicans, and influenza A virus by human neutrophils . Blocking of the interaction between rfSP-D/anti-CD89 and either the pathogen or CD89 abolished its stimulatory effect on pathogen uptake by neutrophils . In addition, rfSP-D/anti-CD89 stimulated killing of E . coli and C . albicans by neutrophils and enhanced neutrophil activation by influenza A virus . In conclusion, rfSP-D/anti-CD89 effectively targeted three structurally unrelated pathogens to neutrophils . (Col)lectin-based chimeric proteins may thus offer promise for therapy of infectious disease.

J Heart Lung Transplant, 2004 Apr, 23(4), 499 - 502
Severe obstruction of the left main coronary artery by mycotic aortic psuedoaneurysm following orthotopic heart transplantation; Kamineni R et al.; Mycotic aneurysm of the ascending aorta is a rare complication following orthotopic heart transplantation . This article describes a case of mycotic pseudoaneurysm caused by Candida albicans that developed shortly after orthotopic heart transplantation . The pseudoaneurysm compressed the left main coronary artery, which led to the development of congestive heart failure symptoms mimicking sub-acute transplant rejection . The heart failure signs and symptoms resolved completely with resection of the aneurysm . This case reiterates that early diagnosis and complete resection of the aneurysm is associated with good prognosis.

Zhonghua Kou Qiang Yi Xue Za Zhi, 2004 Mar, 39(2), 149 - 52
{The attribute of Candida albicans isolates from patients with oral lichen planus}; Zeng X et al.; OBJECTIVE: To investigate the genotypic profiles of Candida albicans isolates from erosive oral lichen planus (OLP) and nonerosive OLP, and then to compare the results with their virulence attributes . METHODS: A total of 112 isolates from healthy control (26), erosive OLP (62) and nonerosive OLP (24) were screened for genotypic profiles by using the randomly amplified polymorphic DNA (RAPD) assay . In addition, adhesion to buccal epithelial cells assay and phospholipase activity assay were used to evaluate the virulence attributes of these isolates . RESULTS: RAPD analyses with some random primer revealed 4 different genotypes among all isolates, and there was significant difference in the geneotypic constitution between every two groups . Statistically, in healthy group the major type was B and D, however, the major type in erosive OLP was A and C, and the major type in nonerosive OLP was A and D . The isolates with genotype A had the strongest adherence among 4 genotypes . The phospholipase activity of the isolates with genotype A and C were higher than that with genotype B and D . CONCLUSIONS: Some Candida albicans isolates with special genotypic profiles and virulence attributes may contribute to the development and progression of OLP.

Immunol Cell Biol, 2004 Apr, 82(2), 196 - 204
Innate versus adaptive immunity in Candida albicans infection; Ashman RB et al.; Candida albicans is a common opportunistic pathogen, causing both superficial and systemic infection . Clinical observations indicate that mucocutaneous infections are commonly associated with defective cell-mediated immune responses, whereas systemic infection is more frequently seen in patients with deficiencies in neutrophil number or function . Analysis of mechanisms of host resistance against gastrointestinal and oral infection in mouse models has demonstrated an absolute dependence on CD4(+) T cells, although clearance also involves phagocytic cells . Both IL-12 and TNF-alpha appear to be important mediators, but mouse strain-dependent variations in susceptibility to infection may be related to T-cell enhancement of production of phagocytic cells by the bone marrow . In murine systemic infection, the role of innate and adaptive responses is less well defined . Studies in immunodeficient and T-cell-depleted mice suggest that clearance of the yeast may be predominantly a function of the innate response, whereas the adaptive response may either limit tissue damage or have the potential to cause immunopathology, depending on the host genetic context in which the infection takes place.

J Oral Pathol Med, 2004 Apr, 33(4), 243 - 5
Facial Candida albicans cellulitis occurring in a patient with oral submucous fibrosis and unknown diabetes mellitus after local corticosteroid injection treatment; Chen HM et al.; Facial cellulitis caused by odontogenic bacterial infection is frequently encountered; however, facial cellulitis caused by Candida albicans infection is rarely found . A patient with oral submucous fibrosis (OSF) and unknown diabetes mellitus (DM) was treated in our out-patient dental clinic by biweekly submucosal injection of 40 mg triamcinolone acetonide into bilateral buccal mucosae plus forced mouth opening performed by the two hands of the clinician . The interincisal distance of the patient improved from 28 to 48 mm after four times of steroid injection . The symptoms and signs of OSF also improved markedly . Unfortunately, facial candidal cellulitis occurred 2 months after the last time of steroid injection treatment . The infection was cured by incision and drainage, intravenous administration of amphotericin B (100 mg once a day for a week), and an appropriate medical control of DM . No recurrence of facial cellulitis was found during the follow-up period of 18 months . To prevent the occurrence of facial cellulitis after a high-dose steroid therapy, some prophylactic procedures should be taken before the initiation of the steroid treatment.

Scand J Infect Dis, 2004, 36(2), 124 - 30
Clinical characteristics, treatment and prognostic factors of candidal meningitis in a teaching hospital in Taiwan; Chen TL et al.; The prognostic factors of candidal meningitis had rarely been studied owing to the rarity of this disease . We identified 17 patients with candidal meningitis at a teaching hospital in Taiwan over a 14-y period and give details of the clinical features, treatment, outcome and prognostic factors of this disease . 10 patients were children (1 was neonate) and 7 were adults . The clinical features and cerebrospinal fluid (CSF) findings were non-specific . The disease was diagnosed > 72 h after hospitalization in 14 (82%) patients . The most frequently isolated species was Candida albicans (65%) . Regimens of therapy included intravenous amphotericin B deoxycholate (AmB) or fluconazole alone, or a combination of intravenous AmB with intrathecal AmB, flucytosine or fluconazole . All indwelling central nervous system devices were externalized or removed . Four adults died of the disease . Three of them had malignancy and had received < 48 h of antifungal therapy . For adult patients with malignancy, early diagnosis and alternative treatment modalities with newer antifungal agents may be needed.

J Prosthodont, 2003 Dec, 12(4), 249 - 54
Effect of surface-charged poly(methyl methacrylate) on the adhesion of Candida albicans; Park SE et al.; PURPOSE: The aim of this study was to investigate the ability of a new surface-modified denture resin to reduce adhesion of Candida albicans to the denture surface . This material has a negative charge incorporated by copolymerization of methacrylic acid to methyl methacrylate . An in vitro system was designed to assess the adhesion of C . albicans to surface-charged poly(methyl methacrylate) (PMMA) surfaces . MATERIALS AND METHODS: Four experimental groups consisted of a control (pure PMMA) and 3 groups of modified PMMA (mPMMA) (95:5, 90:10, and 80:20 MMA:methacrylic acid) . Four resin samples (11 x 5 mm) for each experimental group were made to enable the adhesion assay . Resin samples of varying ratios of methacrylic acid were placed in C . albicans suspensions, and the adherent cells were stained with Gram's crystal violet . The surface area of adherent C . albicans was examined for the entire sample using the computer image at a magnification of x 25 . Percentage of surface area of adherent C . albicans colonies was averaged and compared between the control and each experimental group . Analysis of variance and Bonferroni's multiple-comparisons test were performed on the data . RESULTS: As the ratio of incorporated methacrylic acid to PMMA increased, the surface area of adherent C . albicans decreased . Analysis of data revealed a significant decrease in C . albicans adhesion to the resin blocks (p < 0.0001) when the methacrylic acid was present at 10% of the PMMA . There also existed a correlation between the surface area of adherent C . albicans and the contact angle measurement as the ratio of methacrylic acid changed . Surface contact angle analysis of resin surfaces revealed that as the amount of methacrylic acid increased, the surface energy of the resins increased . CONCLUSIONS: The present results support the role of electrostatic interaction in adhesion, and introduce an effective method of reducing adhesion of C . albicans to PMMA surfaces through modification of the surface charge of polymeric biomaterials.

Pediatrics, 2004 Apr, 113(4), e360 - 4
Raynaud's phenomenon of the nipple: a treatable cause of painful breastfeeding; Anderson JE et al.; Maurice Raynaud first described the vasospasm of arterioles in 1862, and Raynaud's phenomenon is now felt to be common, affecting up to 20% of women of childbearing age . Raynaud's phenomenon has been reported to affect the nipples of breastfeeding mothers and is recognized by many lactation experts as a treatable cause of painful breastfeeding . In 1997, Lawlor-Smith and Lawlor-Smith reported 5 women with Raynaud's phenomenon associated with breastfeeding, but there are few other case reports, and none report the possible relationship between Raynaud's phenomenon of the nipple and previous breast surgery . We report 12 women who breastfed 14 infants, all of whom were seen in 1 pediatric practice and 1 lactation consultation center in San Francisco, California, within the past 3 years . Of the 12 women, 11 were seen between June 2002 and May 2003 . All women suffered from extremely painful breastfeeding, with symptoms precipitated by cold temperatures and associated with blanching of the nipple followed by cyanosis and/or erythema . Poor positioning and poor attachment or latch may cause blanching of the nipple and pain during breastfeeding, but 10 of the 12 mothers were evaluated by experienced lactation consultations, who were sure that inappropriate breastfeeding techniques were not contributing factors . Because the breast pain associated with Raynaud's phenomenon is so severe and throbbing, it is often mistaken for Candida albicans infection . It is not unusual for mothers who have Raynaud's phenomenon of the nipple to be treated inappropriately and often repeatedly for C albicans infections with topical or systemic antifungal agents . Eight of our 12 mothers and their infants received multiple courses of antifungal therapy without relief before the diagnosis was made . To diagnose Raynaud's phenomenon accurately, additional symptoms such as precipitation by cold stimulus, occurrence of symptoms during pregnancy or when not breastfeeding, and biphasic or triphasic color changes must be present . All our mothers experienced precipitation of symptoms by cold stimuli and demonstrated biphasic or triphasic color changes, and 6 of the 12 experienced symptoms during pregnancy . Interestingly 3 of 12 mothers also reported a history of breast surgery, including 1 mother who had a fibroadenoma removed and 2 who had breast-reduction surgery . The association between breast surgery/implants and autoimmune disease, including Raynaud's phenomenon, has been discussed extensively, but the association of Raynaud's phenomenon of the nipple during breastfeeding has not been reported previously . Given the small numbers in the study, it is uncertain as to whether this may be a precipitating factor in developing Raynaud's phenomenon . Treatment options include methods to prevent or decrease cold exposure, avoidance of vasoconstrictive drugs/nicotine that could precipitate symptoms, and pharmacologic measures . There are reports in the lay press of the use of herbal medicines, aerobic exercise, and dietary supplements, but because most women with painful breastfeeding require immediate relief of the pain to continue breastfeeding successfully, it is important to offer a treatment plan that will alleviate the pain quickly . Nifedipine, a calcium channel blocker, has been used to treat Raynaud's phenomenon because of its vasodilatory effects . Very little of the medication can be demonstrated in breast milk and thus is safe to use in breastfeeding mothers . Of the 12 mothers in our series, 6 chose to use nifedipine, and all had prompt relief of pain . Only 1 mother developed side effects from nifedipine . Pediatricians and lactation consultants should be aware of this treatable cause of painful breastfeeding and should specifically question their patients, because most mothers will not provide this information to the breastfeeding consultant . Prompt treatment will allow mothers to continue to breastfeed pain free while avoiding unnecessary antifungal therapy.

Am J Ophthalmol, 2004 Apr, 137(4), 725 - 31
Culture-proven endogenous endophthalmitis: clinical features and visual acuity outcomes; Schiedler V et al.; PURPOSE: To investigate clinical features and visual acuity outcomes associated with endogenous endophthalmitis . DESIGN: Retrospective, observational case series . METHODS: Twenty-one eyes of 21 patients treated at Bascom Palmer Eye Institute for culture-proven endogenous endophthalmitis between 1996 and 2002 were reviewed . RESULTS: Patients were followed a mean of 3 months (range, 1 to 12 months) . Fungal isolates occurred in 13 eyes (62%), gram-positive isolates in 7 (33%), and gram-negative isolates in 1 (5%) . Twelve patients (57%) were hospitalized at the time of diagnosis and 6 patients (29%) died within 2 months of diagnosis . Initial treatment included tap and injection of intravitreal medication in 10 eyes (48%) and pars plana vitrectomy with injection of intravitreal medication in 11 eyes (52%) . Final visual outcomes were obtainable for 18 eyes (two patients died within 10 days of diagnosis, and one patient was lost to follow-up) . Eight (44%) of these 18 eyes achieved a visual acuity of 20/400 or better and 10 (56%) of 18 eyes achieved a visual acuity worse than 20/400, including 3 that were either enucleated or eviscerated . Three eyes with Aspergillus endophthalmitis had worse visual outcomes than eyes with either Candida (P =.036) or bacterial endophthalmitis (P =.024) . CONCLUSIONS: Compared with published series of postoperative or post-traumatic endophthalmitis, patients with endogenous endophthalmitis are more likely to have fungal isolates with a predominance of Candida albicans . Endogenous endophthalmitis is generally associated with high mortality and poor visual acuity outcomes, particularly when caused by more virulent species such as Aspergillus.

Cytometry A, 2004 Apr, 58A(2), 201 - 6
A single-step, sensitive flow cytofluorometric assay for the simultaneous assessment of membrane-bound and ingested Candida albicans in phagocytosing neutrophils; Busetto S et al.; BACKGROUND: Distinguishing ingested particles from those attached to the cell surface is an essential requirement when performing quantitative studies of phagocytosis . In the present report, we describe a simple, sensitive and reliable flow cytofluorometric method that achieves this goal in a Candida albicans-human neutrophils (PMN) system . METHODS: The assay is based on the observation that the vital dye trypan blue (TB), while quenching the green fluorescence of fluorescein-labeled C . albicans, causes them to fluoresce red . PMN were incubated with fluorescein-labeled yeast particles for the required time . Aliquots of the incubation mixtures were then promptly diluted with an equal volume of a TB solution at pH 4.0, and subsequently analyzed by flow cytometry for green and red fluorescence . RESULTS: Since TB does not penetrate into the cells, ingested yeasts retain their green fluorescence, while membrane-bound particles display a red fluorescence . CONCLUSIONS: Our fluorescence flow cytometric method enables to simultaneously distinguish, within the leukocyte population, cell subsets with attached and ingested yeast particles . Its major features are: (1) accuracy, sensitivity and reproducibility; (2) no further sample manipulations after completion of phagocytosis; (3) possibility of counting free, attached and internalized yeast particles; and (4) use of a nontoxic reagent (TB) .

Curr Microbiol, 2004 Apr, 48(4), 312 - 7
Identification and antimicrobial activity of phenylacetic acid produced by Bacillus licheniformis isolated from fermented soybean, Chungkook-Jang; Kim Y et al.; A bacterial strain, B65-1, which showed strong antimicrobial activity, was isolated from Chungkook-Jang, a traditional Korean fermented-soybean food with antimicrobial properties . Based on carbon utilization pattern and partial 16S rRNA sequence analysis, the B65-1 strain was identified as Bacillus licheniformis . An antibiotic compound, active against bacteria and yeast such as Staphylococcus aureus, Escherichia coli, and Candida albicans, was isolated by various chromatographic procedures from culture filtrates of B . licheniformis B65-1 . The purified antibiotic was identified to be phenylacetic acid, with the molecular formula C(8)H(8)O(2) by analyses of EI-MS and NMR . The phenylacetic acid was detected in fermented soybean made with the strain B65-1 as a starter, but was not present in extracts of nonfermented soybean . Our results indicated that the phenylacetic acid produced by B . licheniformis during fermentation of soybean is one of the main compounds of antimicrobial activity of Chungkook-Jang.

J Med Chem, 2004 Apr 8, 47(8), 2133 - 56
Distamycin analogues with enhanced lipophilicity: synthesis and antimicrobial activity; Khalaf AI et al.; Forty-eight heterocyclic amino acid trimers, analogues of distamycin, with a number of features that enhance lipophilicity are described . They contain alkyl or cycloalkyl groups larger than methyl; some are N-terminated by acetamide or methoxybenzamide and are C-terminated by dimethylaminopropyl or aliphatic heterocylic aminopropyl substituents . The ability of these compounds to bind principally to AT tracts of DNA has been evaluated using capillary zone electrophoresis . Significant antimicrobial activity against key organisms such as MRSA and Candida albicans is shown by several compounds, especially those containing a thiazole . Moreover, these compounds have low toxicity with respect to several mammalian cell lines.

Biotechnol Lett, 2004 Feb, 26(4), 337 - 41
Structure and fungicidal activity of a synthetic antimicrobial peptide, P18, and its truncated peptides; Lee DG et al.; P18 (KWKLFKKIPKFLHLAKKF-NH2) is an antimicrobial peptide designed from a cecropin A-magainin 2 hybrid that has potent antibacterial activity without hemolytic activity against human erythrocytes . In this study, P18 displayed potent fungicidal activity (MIC: 12.5 approximately 25 microM) against pathogenic fungi, Candida albicans, Trichosporon beigelii, Aspergillus flavus and Fusarium oxyspovrum . The central Pro9 residue and the entire sequence of P18 are essential for its full fungicidal activity . Circular dichroism analysis suggested that the higher alpha-helical content of the peptides did not correlate with the stronger fungicidal activity.

Ann Otol Rhinol Laryngol, 2004 Mar, 113(3 Pt 1), 225 - 8
Isolated necrotizing epiglottitis: report of a case in a neutropenic patient and review of the literature; Sengor A et al.; Isolated necrotizing epiglottitis (INE) is an unusual condition that may develop in immunocompromised patients . Only 3 cases of INE have been reported in the English-language literature; this is the fourth case, one in a 27-year-old woman who had neutropenia due to cytomegalovirus infection . Stenotrophomonas maltophilia and Candida albicans were isolated from the culture of necrotic material . The features of INE are discussed here by reviewing the literature and by examining the characteristics of this case.

Proc Natl Acad Sci U S A, 2004 Apr 6, 101(14), 5048 - 52 Epub 2004 Mar 29.
Tyrosol is a quorum-sensing molecule in Candida albicans; Chen H et al.; The human fungal pathogen Candida albicans shows a significant lag in growth when diluted into fresh minimal medium . This lag is abolished by the addition of conditioned medium from a high-density culture . The active component of conditioned medium is tyrosol, which is released into the medium continuously during growth . Under conditions permissive for germ-tube formation, tyrosol stimulates the formation of these filamentous protrusions . Because germ-tube formation is inhibited by farnesol, another quorum-sensing molecule, this process must be under complex positive and negative control by environmental conditions . The identification of tyrosol as an autoregulatory molecule has important implications on the dynamics of growth and morphogenesis in Candida.

Eur J Med Chem, 2004 Mar, 39(3), 291 - 8
Synthesis and structure-activity relationships of new antimicrobial active multisubstituted benzazole derivatives; Yildiz-Oren I et al.; A series of multisubstituted benzoxazoles, benzimidazoles, and benzothiazoles (5-7) as non-nucleoside fused isosteric heterocyclic compounds was synthesized and tested for their antibacterial activities against various Gram-positive and Gram-negative bacteria and antifungal activity against the fungus Candida albicans . Microbiological results indicated that the synthesized compounds possessed a broad spectrum of activity against the tested microorganisms at MIC values between 100 and 3.12 microg/ml . Structure-activity relationships (SAR) studies revealed that benzothiazole ring system enhanced the antimicrobial activity against Staphylococcus aureus . In these sets of non-nucleoside fused heterocyclic compounds electron withdrawing groups at position 5 of the benzazoles increased the activity against C . albicans.

Bioorg Med Chem, 2004 Apr 15, 12(8), 1935 - 45
Design, synthesis and biological evaluation of some pyrazole derivatives as anti-inflammatory-antimicrobial agents; Bekhit AA et al.; The synthesis of novel series of structurally related 1H-pyrazolyl derivatives is described . All the newly synthesized compounds were tested for their in vivo anti-inflammatory activity by two different bioassays namely; cotton pellet-induced granuloma and sponge implantation model of inflammation in rats . In addition, COX-1 and COX-2 inhibitory activities, ulcerogenic effects and acute toxicity were determined . The same compounds were evaluated for their in vitro antimicrobial activity against Escherichia coli, as an example of Gram negative bacteria, Staphylococcus aureus as an example of Gram positive bacteria, and Candida albicans as a representative of fungi . The combined anti-inflammatory data from local and systemic in vivo animal models showed that compounds 4, 5, 8, 9, 11 and 12a exhibited anti-inflammatory activity comparable to that of indomethacin with no or minimal ulcerogenic effects and high safety margin (LD(50)>500 mg/Kg) . In addition, compounds 4, 7, 10, 12a and 12b displayed appreciable antibacterial activities when compared with ampicillin, especially against S . aureus . Compounds 4 and 12a are the most distinctive derivatives identified in the present study because of their remarkable in vivo and in vitro anti-inflammatory potency and their pronounced antibacterial activities comparable to ampicillin against Gram positive . On the other hand, compound 12a exhibited good selective inhibitory activity against COX-2 enzyme . Therefore, such compound would represent a fruitful matrix for the development of anti-inflammatory-antimicrobial candidates.

Fungal Genet Biol, 2004 May, 41(5), 553 - 62
Genetic evidence for recombination in Candida albicans based on haplotype analysis; Tavanti A et al.; The possibility of sexual reproduction in the human pathogenic fungus Candida albicans is a question of great interest in medical mycology . Not only is it a fundamental biological issue, but it is also a potential mechanism for contributing to the phenotypic plasticity (and hence the virulence) of the organism . Molecular genotyping methods such as multi-locus sequence typing (MLST) are generating data that can shed light on this question . In the present study we have used MLST information to generate haplotypes that identify many different homologues of a chromosome within a collection of strains . Particular combinations of these haplotypes provide evidence for chromosomal segregation and intra-chromosome recombination . All of our observations of haplotype diversity could also be explained by other mechanisms, such as gene conversion or mitotic recombination, and the resolution of these issues will require a denser map of accurately localised markers . A common event observed in strain evolution is loss of heterozygosity at a particular marker . Our results contribute to the emerging picture of C . albicans as an organism whose primary means of reproduction is clonal, but with a small but important contribution from sexual reproduction, occurring in nature but not under commonly used laboratory conditions.

Mol Microbiol, 2004 Apr, 52(1), 141 - 58
Has1p, a member of the DEAD-box family, is required for 40S ribosomal subunit biogenesis in Saccharomyces cerevisiae; Emery B et al.; The Has1 protein, a member of the DEAD-box family of ATP-dependent RNA helicases in Saccharomyces cerevisiae, has been found by different proteomic approaches to be associated with 90S and several pre-60S ribosomal complexes . Here, we show that Has1p is an essential trans-acting factor involved in 40S ribosomal subunit biogenesis . Polysome analyses of strains genetically depleted of Has1p or carrying a temperature-sensitive has1-1 mutation show a clear deficit in 40S ribosomal subunits . Analyses of pre-rRNA processing by pulse-chase labelling, Northern hybridization and primer extension indicate that these strains form less 18S rRNA because of inhibition of processing of the 35S pre-rRNA at the early cleavage sites A0, A1 and A2 . Moreover, processing of the 27SA3 and 27SB pre-rRNAs is delayed in these strains . Therefore, in addition to its role in the biogenesis of 40S ribosomal subunits, Has1p is required for the optimal synthesis of 60S ribosomal subunits . Consistent with a role in ribosome biogenesis, Has1p is localized to the nucleolus . On sucrose gradients, Has1p is associated with a high-molecular-weight complex sedimenting at positions equivalent to 60S and pre-60S ribosomal particles . A mutation in the ATP-binding motif of Has1p does not support growth of a has1 null strain, suggesting that the enzymatic activity of Has1p is required in ribosome biogenesis . Finally, sequence comparisons suggest that Has1p homologues exist in all eukaryotes, and we show that a has1 null strain can be fully complemented by the Candida albicans homologue.

Mem Inst Oswaldo Cruz, 2003 Dec, 98(8), 1027 - 32 Epub 2004 Mar 09.
Use of molecular methods in identification of Candida species and evaluation of fluconazole resistance; Cirak MY et al.; The aim of this study was to evaluate the use of one of the molecular typing methods such as PCR (polymerase chain reaction) following by RFLP (restriction fragment length polymorphism) analysis in the identification of Candida species and then to differentiate the identified azole susceptible and resistant Candida albicans strains by using AP-PCR (arbitrarily primed-polymerase chain reaction) . The identification of Candida species by PCR and RFLP analysis was based on the size and primary structural variation of rDNA intergenic spacer regions (ITS) . Forty-four clinical Candida isolates comprising 5 species were included to the study . The amplification products were digested individually with 3 different restriction enzymes: HaeIII, DdeI, and BfaI . All the isolates tested yielded the expected band patterns by PCR and RFLP analysis . The results obtained from this study demonstrate that Candida species can be differentiated as C . albicans and non-C . albicans strains only by using HaeIII restriction enzyme and BfaI maintains the differentiation of these non-C . albicans species . After identification Candida species with RFLP analysis, C . albicans strains were included to the AP-PCR test . By using AP-PCR, fluconazole susceptible and resistant strains were differentiated . Nine fluconazole susceptible and 24 fluconazole resistant C . albicans were included to the study . Fluconazole resistant strains had more bands when evaluating with the agarose gel electrophoresis but there were no specific discriminatory band patterns to warrant the differentiation of the resistance . The identification of Candida species with the amplification of intergenic spacer region and RFLP analysis is a practical, short, and a reliable method when comparing to the conventional time-consuming Candida species identification methods . The fluconazole susceptibility testing with AP-PCR seems to be a promising method but further studies must be performed for more specific results.

Proteomics, 2004 Apr, 4(4), 1204 - 15
Contribution of the antibodies response induced by a low virulent Candida albicans strain in protection against systemic candidiasis; Fernandez-Arenas E et al.; A low virulent Candida albicans mutant, CNC13, deleted in the Mitogen Activated Protein (MAP) kynase HOG1 was used to immunize BALB/c mice . Hog1p is essential for the oxidative stress and hyperosmolarity responses . Several doses and immunization procedures were employed . The protection capacity of the different sera generated was analyzed in a murine model of systemic candidiasis . Using a proteomic approach (two-dimensional gel electrophoresis followed by Western blotting), we were able to distinguish two categories of serum: protective and nonprotective, which showed different titres of total Immunoglobulins (Igs) and IgG2a (analyzed by enzyme-linked immunosorbent assay) . The levels of Igs and IgG2a in protective sera were significantly higher compared to nonprotective sera . The pattern of a "nonprotective" profile was composed of enolase (Eno1p), transketolase, heat shock protein and methionine synthase . Only antibodies against enolase are the IgG2a isotype . The pattern of a "protective" sera, on the other hand, was composed of antibodies against the following antigens: several isoforms of Eno1p, pyruvate decarboxylase, pyruvate kynase, a protein of the 40S ribosomal subunit, triosephosphate isomerase, DL-glycerol phosphatase and fructose-bisphosphate aldolase . All these antibodies are the IgG2a isotype . The proteins described in the protective sera might be useful for future vaccine development.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1382 - 3
Caspofungin resistance in Candida albicans: correlating clinical outcome with laboratory susceptibility testing of three isogenic isolates serially obtained from a patient with progressive Candida esophagitis; Hernandez S et al.; A patient with azole-refractory thrush-esophagitis responded initially to caspofungin, but the treatment eventually failed . In a murine model, caspofungin was effective against two early isolates for which the MICs of caspofungin were low, but it was less effective against a late isolate for which the MIC of caspofungin was greater . We concluded that there is a correlation between in vivo failure and rising in vitro caspofungin MICs.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1256 - 71
Chemosensitization of fluconazole resistance in Saccharomyces cerevisiae and pathogenic fungi by a D-octapeptide derivative; Niimi K et al.; Hyperexpression of the Saccharomyces cerevisiae multidrug ATP-binding cassette (ABC) transporter Pdr5p was driven by the pdr1-3 mutation in the Pdr1p transcriptional regulator in a strain (AD/PDR5(+)) with deletions of five other ABC-type multidrug efflux pumps . The strain had high-level fluconazole (FLC) resistance (MIC, 600 microg ml(-1)), and plasma membrane fractions showed oligomycin-sensitive ATPase activity up to fivefold higher than that shown by fractions from an isogenic PDR5-null mutant (FLC MIC, 0.94 microg ml(-1)) . In vitro inhibition of the Pdr5p ATPase activity and chemosensitization of cells to FLC allowed the systematic screening of a 1.8-million-member designer D-octapeptide combinatorial library for surface-active Pdr5p antagonists with modest toxicity against yeast cells . Library deconvolution identified the 4-methoxy-2,3,6-trimethylbenzensulfonyl-substituted D-octapeptide KN20 as a potent Pdr5p ATPase inhibitor (concentration of drug causing 50% inhibition of enzyme activity {IC(50)}, 4 microM) which chemosensitized AD/PDR5(+) to FLC, itraconazole, and ketoconazole . It also inhibited the ATPase activity of other ABC transporters, such as Candida albicans Cdr1p (IC(50), 30 microM) and Cdr2p (IC(50), 2 microM), and chemosensitized clinical isolates of pathogenic Candida species and S . cerevisiae strains that heterologously hyperexpressed either ABC-type multidrug efflux pumps, the C . albicans major facilitator superfamily-type drug transporter Ben(R)p, or the FLC drug target lanosterol 14 alpha-demethylase (Erg11p) . Although KN20 also inhibited the S . cerevisiae plasma membrane proton pump Pma1p (IC(50), 1 microM), the peptide concentrations required for chemosensitization made yeast cells permeable to rhodamine 6G . KN20 therefore appears to indirectly chemosensitize cells to FLC by a nonlethal permeabilization of the fungal plasma membrane.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1242 - 8
Modulation of in vitro fungicidal activity of human lactoferrin against Candida albicans by extracellular cation concentration and target cell metabolic activity; Viejo-Diaz M et al.; The anti-Candida activity of the innate defense protein human lactoferrin was investigated . Lactoferrin displayed a clear fungicidal effect against Candida albicans only under low-strength conditions . This candidacidal activity was inversely correlated with the extracellular concentration of the monovalent cations and was prevented by Na(+) and K(+) (> or 30 mM) and by divalent cations (Ca(2+) and Mg(2+) at > or 4 mM) . A slight cellular release of K(+), cytosolic acidification, and a change in the membrane potential were observed in C . albicans cells treated with lactoferrin, suggesting that this protein directly or indirectly interacts with the cytoplasmic membrane . Mitochondrial inhibitors (carbonyl cyanide m-chlorophenylhydrazone, 2,4-dinitrophenol, azide, and antimycin) as well as anaerobic conditions significantly reduced the killing effect of lactoferrin . These results suggest that low-strength conditions and the cellular metabolic state may modulate the candidacidal activity of human lactoferrin.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1136 - 44
The Candida albicans lanosterol 14-alpha-demethylase (ERG11) gene promoter is maximally induced after prolonged growth with antifungal drugs; Song JL et al.; The azole antifungal drugs that target lanosterol 14-alpha-demethylase, encoded by the ERG11 gene, are used to treat a variety of infections caused by Candida albicans . Azoles are known to induce expression of ERG11 mRNA . The ERG11 promoter was cloned 5' of the luciferase-coding region, and the induction of ERG11 expression by azoles was monitored by luciferase assays . Maximal induction of the ERG11 promoter by azoles occurs not during logarithmic growth but after the diauxic shift and requires azoles to be present throughout logarithmic growth . The effects of pH, carbon source, and aerobic or anaerobic growth on induction of the ERG11 promoter by azoles were analyzed . Treatment with terbinafine and fenpropimorph, which target other enzymes in the ergosterol biosynthetic pathway, also resulted in a delayed induction of ERG11 promoter activity . Nascent sterol synthesis was shown to parallel ERG11 promoter activity, and total sterols were reduced coincident with the timing of ERG11 promoter activation . These results as a whole suggest that expression of the ERG11 promoter is regulated in response to sterol depletion.

J Nat Prod, 2004 Mar, 67(3), 503 - 5
New nitrogenous Germacranes from a Thai marine sponge, Axinyssa n . sp; Satitpatipan V et al.; Two new germacrane sesquiterpenes, including (1Z,4Z)-7alphaH-11-aminogermacra-1(10),4-diene (1) and N,N'-11-bis{(1Z,4Z)-7alphaH-germacra-1(10),4-dienyl}urea (2), were isolated from a Thai marine sponge Axinyssan . sp . collected from the Andaman Sea . The structure elucidations of 1 and 2 were accomplished by means of 1D and 2D NMR, MS, and IR spectroscopy . Only 1 exhibited strong antimicrobial activity against Staphylococcus aureus, Bacillus subtilis, and Candida albicans.

Yeast, 2004 Mar, 21(4), 333 - 40
GFP as a quantitative reporter of gene regulation in Candida albicans; Barelle CJ et al.; A system has been developed for the quantitative analysis of gene expression within individual Candida albicans cells in infected tissue . The system is based on the plasmid pGFP, which contains the codon-optimized yeast enhanced green fluorescent protein (yEGFP; Cormack et al., 1997) cloned between a basal CaADH1 promoter and the ScCYC1 terminator on an integrating vector . Promoters were inserted into pGFP and GFP levels measured in individual cells by quantitative fluorescence microscopy . Analysis of pPCK1-GFP and pMET3-GFP fusions revealed that GFP folds rapidly following gene induction, and is turned over rapidly following gene repression . Hence, single cell fluorescence measurements are likely to reflect ongoing gene expression levels with reasonable accuracy . pACT1-GFP expression levels were relatively constant during growth of C . albicans in both yeast and hyphal forms, and during growth in vivo in the mouse model of systemic infection . Therefore, pACT1-GFP provides a useful control for this quantitative GFP-based system in future analyses of C . albicans molecular responses during fungal infections .

Yeast, 2004 Mar, 21(4), 285 - 302
Identification of potential cell-surface proteins in Candida albicans and investigation of the role of a putative cell-surface glycosidase in adhesion and virulence; Alberti-Segui C et al.; Cell-surface proteins are attractive targets for the development of novel antifungals as they are more accessible to drugs than are intracellular targets . By using a computational biology approach, we identified 180 potential cell-surface proteins in Candida albicans, including the known cell-surface adhesin Als1 and other cell-surface antigens, such as Pra1 and Csa1 . Six proteins (named Csf1-6 for cell-surface factors) were selected for further biological characterization . First, we verified that the selected CSF genes are expressed in the yeast and/or hyphal form and then we investigated the effect of the loss of each CSF gene on cell-wall integrity, filamentation, adhesion to mammalian cells and virulence . As a result, we identified Csf4, a putative glycosidase with an apparent orthologue in Saccharomyces cerevisiae (Utr2), as an important factor for cell-wall integrity and maintenance . Interestingly, deletion of CSF4 also resulted in a defect in filamentation, a reduction in adherence to mammalian cells in an in vitro adhesion assay, and a prolongation of survival in an immunocompetent mouse model of disseminated candidiasis . A delay in colonization of key organs (e.g . kidney) was also observed, which is consistent with a reduction in virulence of the csf4-deletion strain . These data indicate a key role for extracellular glycosidases in fungal pathogenesis and represent a new site for therapeutic intervention to cure and prevent fungal disease .

Cancer Detect Prev, 2004, 28(1), 17 - 26
Phagocytosis of Candida albicans by metastatic and non metastatic human breast cancer cell lines in vitro; Ghoneum M et al.; Experiments were carried out to investigate the kinetic characteristics of phagocytosis of candida by metastatic (MCF-7 and ZR-75-1) and nonmetastatic (HCC70) breast cancer cell (BCC) lines . Cancer cells were mixed with candida at a ratio of 1:10 and attachment/phagocytosis were examined using cytospin preparations stained with either Giemsa or tannic acid and May-Grunwald Giemsa and by using flow cytometry . A high attachment of candida to cancer cells (29-39%) was detected as early as 10 min . Following attachment, cancer cells phagocytized yeast . The phagocytic activity of MCF-7 and ZR-75 cells was significantly higher (58-61%) than that of HCC70 cells (26%) . A similar trend was observed with respect to the phagocytic index . Phagocytosis of candida by tumor cells was inhibited significantly by both cytochalasin B (50%) and by lowering temperature to 4 degrees C (66%) . Phagocytosis was not associated with oxidative burst in any cell lines used . In conclusion, metastatic breast cancer cells possess phagocytic activity which may explain their invasive property.

Infect Immun, 2004 Apr, 72(4), 2390 - 4
SKN7 of Candida albicans: mutant construction and phenotype analysis; Singh P et al.; The SKN7 two-component response regulator gene of Candida albicans was deleted, and the phenotype of the mutant was established . This mutant exhibited impaired growth on Spider agar and 10% serum agar compared to wild-type and gene-reconstituted strains . The skn7 mutant was sensitive to H(2)O(2) in vitro, but its virulence was only mildly attenuated . A comparison of the Skn7p and Ssk1p response regulators of C . albicans is discussed.

Infect Immun, 2004 Apr, 72(4), 2386 - 9
Tec1p-independent activation of a hypha-associated Candida albicans virulence gene during infection; Staib P et al.; The Tec1p transcription factor is involved in the expression of hypha-specific genes in Candida albicans . Although the induction of the hypha-associated SAP5 gene by serum in vitro depends on Tec1p, deletion of all Tec1p binding site consensus sequences from the SAP5 promoter did not affect its activation . In two different animal models of candidiasis, the SAP5 promoter was induced even in a Deltatec1 deletion mutant, demonstrating that the requirement for Tec1p in gene expression in C . albicans depends on the environmental conditions within the host.

What Is MIC?, What Is Bioassay?, What Is Activated Sludge?, What Is Bioengineering?, What Is Bioreactor?, o, Microorganisms, n, Bacterium, e, Microbe, i, Microbes, r, Microorganism, o, Microorganism, a, Escherichia coli, s, Bacteria, c, Antibiotics, a, Antibiotics, e, Streptococcal, c, Cell suspensions, s, Microorganisms, c, Escherichia coli, s, Streptococci, a, Escherichia coli, a, Streptococci, s, Biological reactors, e, Escherichia coli, i, Thermophiles, n, Microorganisms, c, Lactobacillus, o, Clostridia, e, Antibiotics, e, Sepsis, n, Escherichia coli

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005