Candida albicans

Mol Biol Evol, 2003 May, 20(5), 772 - 4 Epub 2003 Apr 02.
Rate of protein evolution versus fitness effect of gene deletion; Yang J et al.; Whether nonessential genes evolve faster than essential genes has been a controversial issue . To resolve this issue, we use the data from a nearly complete set of single-gene deletions in the yeast Saccharomyces cerevisiae to assess protein dispensability . Also, instead of the nematode, which was used previously but is only distantly related to S . cerevisiae, we use another yeast, Candida albicans, as a second species to estimate the evolutionary distances between orthologous genes in two species . Our analysis reveals only a weak correlation between protein dispensability and evolutionary rate . More important, the correlation disappears when duplicate genes are removed from the analysis . And surprisingly, the average rate of nonsynonymous substitution is considerably lower than that for single-copy genes in the yeast genome . This observation suggests that structural constraints are more important in determining the rate of evolution of a protein than dispensability because duplicate genes are on average more dispensable than single-copy genes . For duplicate genes, those with only a weak effect or no effect of deletion on fitness evolve on average faster than those with a moderate or strong effect of deletion on fitness, which in turn evolve on average faster than those with a lethal effect of deletion.

J Wound Care, 2003 Mar, 12(3), 101 - 7
A comparison of the antimicrobial effects of four silver-containing dressings on three organisms; Thomas S et al.; OBJECTIVE: To compare the antimicrobial activity of four silver-containing dressings: Acticoat (Smith and Nephew), Actisorb Silver 220 (Johnson and Johnson), Avance (SSL International) and Contreet-H (Coloplast) . METHOD: The dressings were tested against three micro-organisms: a Gram-positive bacterium, Staphylococcus aureus; a Gram-negative organism, Escherichia coli; and a yeast, Candida albicans, using three different techniques . Each dressing also had its silver content analysed . RESULTS: Acticoat is likely to produce the most rapid antimicrobial effect in vitro because of the rapid release of relatively large concentrations of highly active silver ions . Contreet-H has broadly similar antimicrobial activity to Acticoat, but has a slower onset of action . Actisorb Silver 220 appears to offer less prospect of killing bacteria within the wound itself, but the dressing is capable of removing micro-organisms from wound exudate and sequestering them until they are inactivated by the silver within the charcoal fibres . Little convincing evidence for any antimicrobial activity of Avance was found . CONCLUSION: Although caution must always be exercised when extrapolating the results of laboratory-based studies to the clinical situation, potentially important differences were detected in the antimicrobial activity of the four products examined . It is also possible that the silver ions released by the dressings may have effects on wound healing that are unrelated to their antimicrobial activity . Further work is needed to address this issue.

J Med Microbiol, 2003 Apr, 52(Pt 4), 349 - 59
'Genotypic shuffling' of sequential clones of Candida albicans in HIV-infected individuals with and without symptomatic oral candidiasis; Samaranayake YH et al.; Although HIV-infected individuals harbour multiple strains of oral Candida albicans, little is known of their micro-evolution over time . Therefore, a prospective study was conducted with 16 HIV-infected ethnic Chinese individuals with and without symptoms of oropharyngeal candidiasis to evaluate the genotype distribution of oral C . albicans isolates during HIV disease progression . Oral-rinse samples were obtained from all individuals and up to five C . albicans colonies were selected for each visit, over a 12 month period of multiple visits . After identification of isolates using standard mycological criteria, the genetic similarities of yeast isolates within and between sequential clones of C . albicans were assessed by DNA fingerprinting through random amplification of polymorphic DNA (RAPD) . The results of RAPD gel profiles and the lineage of each isolate were further analysed using commercially available software . RAPD studies revealed the prevalence of up to 14 different genotypes per individual during the study period, with multiple genotypes isolated simultaneously from a single oral rinse . Computer analysis of RAPD profiles revealed that yeasts isolated over sequential visits from symptomatic individuals demonstrated a striking level of relatedness compared with isolates from asymptomatic individuals . Genetically identical C . albicans strains also formed 'loosely' connected subclusters that overlapped multiple visits, implying genetic 'shuffling' in these isolates during disease progression . These data point to varying evolutionary genetic trends in C . albicans associated with symptomatic oral candidiasis and asymptomatic carriage in HIV disease.

Comp Immunol Microbiol Infect Dis, 2003 Jul, 26(4), 223 - 32
Susceptibility to fungal infections of nails in patients with primary antibody deficiency; Macura AB et al.; Primary antibody deficiencies are rare diseases, which require early treatment with intravenous immunoglobulins to prevent fatal infections . The cell mediated immunity in patients with those immunodeficiencies remains unimpaired and usually they do not develop fungal infections . The aim of the study was to determine the susceptibility to fungal infections of nails in children with X-linked agammaglobulinaemia (XLA) and common variable immunodeficiency (CVID) . Nail plate fragments collected from five patients with XLA and five with CVID were experimentally infected with a Candida albicans and Trichophyton mentagrophytes strains . The same procedures were carried out with the nails from a control group of 10 healthy volunteers . The intensity of the infection was evaluated on the basis of hyphae ingrown into the nail fragments . The main finding of the study was the increased susceptibility of antibody deficient patients to experimental nail infection with C . albicans and T . mentagrophytes.

Mol Microbiol, 2003 Apr, 48(2), 523 - 36
The regulation of EFG1 in white-opaque switching in Candida albicans involves overlapping promoters; Lachke SA et al.; EFG1, which encodes a trans-acting factor, is expressed as a more abundant 3.2 kb transcript in the white phase and as a less abundant 2.2 kb transcript in the opaque phase of the white-opaque transition in Candida albicans . To understand how alternative phase-specific mRNAs are transcribed from the same gene locus, the 2320 bp upstream region of the gene was functionally characterized by analysing the -activity of deletion derivatives in a luciferase-based reporter system . The white phase-specific promoter contained three discrete sequences involved in white phase-specific activation, between -2022 and -1809 bp (AR1), between -1809 and -1727 bp (AR2) and between -922 and -840 bp (AR3) . A higher resolution deletion and mutation analysis of AR2 revealed two regions between -1809 and -1787 bp and between -1764 and -1728 bp that are responsible for AR2 activation . Targeting of promoter constructs to the ectopic ADE2 genomic site and the 3' end of the EFG1 genomic site revealed a positional requirement for white phase-regulated activation specific for the AR2 region of the promoter . Gel mobility shift assays using AR2 revealed a white phase-specific activation complex . No discrete activation sequences were identified in the overlapping promoter of the opaque phase-specific EFG1 transcript . The strength of opaque phase activation was directly proportional to the length of the promoter . Northern analysis excluded the possibility of an opaque phase-specific repressor . These results demonstrate overlapping promoters for white and opaque phase-specific expression of the gene for the transcription factor Efg1, with distinctly different mechanisms of phase-specific activation.

Biol Chem, 2003 Jan, 384(1), 183 - 90
Internalisation and degradation of histatin 5 by Candida albicans; Ruissen AL et al.; Histatins, salivary antimicrobial peptides, are susceptible to proteolytic degradation, often ascribed to host proteinases . In this study, we addressed the question whether proteolytic activity from microbial sources can contribute to this degradation . Candida albicans, an opportunistic yeast that is susceptible to the histatins, was used as target organism . The most potent histatin (histatin 5: sequence: DSHAKRHHGYKRKFHEKHHSHRGY), two histatin 5 fragments (dh-5: sequence: KRKFHEKHHSHRGY; P-113: sequence: AKRHHGYKRKFH) and an all-D isomer of the latter (P-113D) were used as model peptides . All L-peptides were susceptible to degradation by C . albicans . Cleavage was established at Lys5 and His19 of histatin 5, Lys11, Arg12, Phe14, Glu16, Lys17, His18 and Ser20 of dh-5 and Ala4 and Lys11 of P-113 . In addition, it was found that secreted C . albicans enzymes are not involved in the degradation process and that blocking cell entry of the peptides greatly impedes degradation . Moreover, P-113D, which is biologically as active as P-113, was hardly susceptible to proteolysis . These data imply that proteolysis occurs mainly intracellularly and is not used as a protective mechanism against histatin activity . Together, our results suggest that, besides host proteinases, microbial enzymes play an important role in histatin degradation.

Yeast, 2003 Apr 15, 20(5), 417 - 26
Characterization of a disulphide-bound Pir-cell wall protein (Pir-CWP) of Yarrowia lipolytica; Jaafar L et al.; In this work we have studied the disulphide-bound group of cell wall mannoproteins of Yarrowia lipolytica and Candida albicans . In the case of Y . lipolytica, SDS-PAGE analysis of the beta-mercaptoethanol-extracted material from the purified cell walls of the yeast form, showed the presence of a main polypeptide of 45 kDa and some minor bands in the 100-200 kDa range . This pattern of bands is similar to that obtained in identical extracts in Saccharomyces cerevisiae (Moukadiri et al., 1999), and besides, all these bands cross-react with an antibody raised against beta-mercaptoethanol-extracted material from the purified cell walls of S . cerevisiae, suggesting that the 45 kDa band could be the homologue of Pir4 of S . cerevisiae in Y . lipolytica . To confirm this possibility, the amino-terminal sequences of two internal regions of the 45 kDa protein were determined, and degenerate oligonucleotides were used to clone the gene . The gene isolated in this way codes a 286 amino acid polypeptide that shows homology with the Pir family of proteins of S . cerevisiae (Russo et al., 1992; Toh-e et al., 1993), accordingly we have named this gene YlPIR1 . Disruption of YlPIR1 led to a slight increase in the resistance of the cells to calcofluor white, Congo red and zymolyase, but did not cause changes in cell morphology, growth rate or morphological transition .

FEMS Microbiol Lett, 2003 Mar 28, 220(2), 247 - 53
Binding, internalisation and degradation of histatin 3 in histatin-resistant derivatives of Candida albicans; Fitzgerald DH et al.; The antifungal mechanism of salivary histatin has been studied in Candida albicans and involves binding to a specific receptor, translocation across the membrane and targeting intracellularly . Cell death correlates with non-lytic release of ATP that may function as a cytotoxic mediator extracellularly . By sequential exposure to increasing concentrations of histatin 3, we generated histatin-resistant derivatives of C . albicans strain CA132A that show five-fold less killing at physiological concentrations of histatin 3 . Protection against histatin killing in histatin-resistant derivatives is not due to alterations in binding, internalisation or degradation of histatin or efflux of ATP . These results indicate that protective mechanisms activated by exposure to histatin 3 may involve unidentified pathways downstream of binding and internalisation events.

Microb Pathog, 2003 Apr, 34(4), 169 - 77
Candida albicans triggers interleukin-8 secretion by oral epithelial cells; Dongari-Bagtzoglou A et al.; Oropharyngeal candidiasis is a frequent opportunistic infection associated with immunocompromised hosts . Candida albicans is the principal species responsible for this infection . Production of interleukin-8 (IL-8), by oral epithelial cells can be expected to play a major role in the recruitment and activation of professional phagocytes at the infected site . The purpose of this study was to determine whether C . albicans triggers secretion of IL-8 by oral epithelial cells in vitro and investigate mechanisms of host cell-fungal interactions that trigger such responses . Oral epithelial cell lines (SCC4, SCC15, and OKF6/TERT-2) as well as primary gingival epithelial cells were used . Epithelial cells were cocultured with C . albicans, strains SC5314, ATCC28366 or ATCC32077, for 24-48 hr, and supernatants were analyzed for IL-8 content by ELISA . A germination-deficient mutant (efg1/efg1 cph1/cph1), otherwise isogenic to strain SC5314, was used to assess the requirement for germination in triggering IL-8 responses . In order to ascertain whether direct contact of yeast with host cells is required to trigger cytokine production, epithelial cells were separated from yeast using cell culture inserts . To test whether IL-8 secretion is dependent on IL-1alpha activity, epithelial cells were challenged with viable C . albicans in the presence or absence of neutralizing anti-IL-1alpha antibody or IL-1ra, and IL-8 secretion was measured in the supernatants . All cell lines and primary cultures responded to C . albicans with an increase in IL-8 secretion . IL-8 responses were contact-dependent, strain-specific, required yeast viability and germination into hyphae, and were in part autoregulated by IL-1alpha .

Regul Pept, 2003 Apr 15, 112(1-3), 147 - 52
Adrenomedullin and mucosal defence: interaction between host and microorganism; Allaker RP et al.; Many surface epithelial cells express adrenomedullin (AM) and it is postulated that it may have an important protective role . This peptide has many properties in common with other cationic antimicrobial peptides including the human beta-defensins . Antimicrobial activity against members of the human skin, oral, respiratory tract and gastric microflora has been demonstrated . Both pathogenic and commensal strains of bacteria are sensitive; Gram-positive and Gram-negative bacteria being equally susceptible . No activity against the yeast Candida albicans was observed . Minimum inhibitory and minimum bacteriocidal concentrations range from 7.75 x 10(-4) to 12.5 and 0.003 to >25.0 microg ml(-1), respectively . On exposure of oral, skin and gastric epithelial cells to whole cells and culture supernatants from bacteria isolated from these sites an increase in AM peptide and gene expression has been observed . No upregulation was detected with C . albicans . In cultured cells and an animal infection model increased AM peptide and gene expression has been demonstrated using immunohistochemical and in situ hybridization techniques . These collective findings suggest that AM represents a new category of antimicrobial peptide, which contributes to the mucosal host defence system.

Mikrobiol Z, 2002 Nov-Dec, 64(6), 57 - 61
{Investigation of the activity of the preparation cerbiden against Candida spp.}; Hladun NP et al.; Antifungal activity of a new complex antibiotical preparation cerbiden obtained from medical plant Bidens cernua L . of the Asteraceae family, was investigated in vitro against the clinical and museum strains of Candida spp . High cerbiden activity against clinical and museum strains of Candida albicans, C . tropicalis, C . krusei, C . parapsilosis, C . guilliermondii, sensitive and resistant to nystatin, amphotericin B and clotrimazole was determined.

Immunol Lett, 2003 Feb 3, 85(3), 251 - 5
Candida albicans mannan-protein conjugate as vaccine candidate; Bystricky S et al.; Mannan-branched polysaccharide with alpha-(1-->6)-linked mannose residues in the linear backbone, is a major virulence and protective factor of yeast Candida albicans . Injected alone does not induce sufficient level of protective antibodies . In this study, we have conjugated mannan to protein carrier by simple one step reaction using 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) activation reagent . Prepared mannan conjugate is immunogenic in rabbits and reinjection elicited a booster response with significant increase of serum IgG level . Anti-C . albicans effectiveness of immune serum is clearly demonstrated . Based on these results, the mannan conjugate synthesized by this scheme can be considered as actual vaccine candidate for clinical evaluation.

Ann Clin Lab Sci, 2003 Winter, 33(1), 86 - 93
Isolation and partial characterization of Hsp90 from Candida albicans; Burt ET et al.; Hsp90 is a stress-induced protein involved in many cellular processes including the regulation of signal transduction and steroid hormone response pathways in higher eukaryotic cells . Candida albicans hsp90 has a mass of 82 -Da and has previously been implicated as a virulence factor . A 47-kDa C-terminal fragment of Candida hsp90 is a target for an immune response to C . albicans infections . A C . albicans hsp90 specific polyclonal antibody was developed against a synthetic peptide containing a previously defined epitope of the 47-kDa fragment . This antibody was used to investigate the cellular localization and induction of hsp90 in the fungus . By means of cell surface protein extraction, hsp90 is shown to be localized on the cell surface as well as in the cytoplasm . On the cell surface, it appears only as an 82-kDa protein . In the cytoplasm, anti-hsp90 detected the 82-kDa protein as well as 72-kDa and 47-kDa bands on SDS-PAGE gels . The cytoplasmic protein bands were heat inducible and appeared to be estrogen induced as well, suggesting that C . albicans modulates hsp90 expression in response to environmental changes . Since the 82-kDa protein is also found on the surface of the cells, hsp90 may be directly involved in sensing environmental changes . It may also be important for recognition of its host or elements of the host immune system and antibody responses to the molecule and may therefore be useful for diagnostic or prognostic evaluation.

Int J Clin Pract, 2003 Mar, 57(2), 143 - 4
Caspofungin in the treatment of oropharyngeal candidiasis; Garbino J et al.; A patient with AIDS developed oropharyngeal candidiasis . Candida albicans and C . glabrata were isolated from the patient and found to be resistant to fluconazole and itraconazole in vitro . Voriconazole therapy was initiated, but discontinued when the C . albicans strain isolated from the patient was found to be resistant to it . The patient failed to respond to subsequent therapy with a combination of amphotericin B and 5-flucytosine . Therapy with caspofungin was then initiated (70 mg loading dose, followed by 50 mg/day) . The patient responded favourably to caspofungin, with complete resolution of signs and symptoms.

Pharmacology, 2003 May, 68(1), 9 - 16
Effect of butylated hydroxyanisole and some of its derivatives on human neutrophil oxidative burst: chemiluminescence evaluation; Braga PC et al.; An acute inflammatory response begins during the reperfusion phase following an ischemic insult in which polymorphonuclear neutrophils (PMNs) play an important role and the release of reactive oxygen species (ROS) causes further damage and a reduction in endogenous antioxidant storage . The ability of butylated hydroxyanisole (BHA) and some phenolic, aliphatic and aromatic BHA derivatives to reduce the human PMN oxidative burst evoked by particulate (Candida albicans and zymosan) or soluble stimulants {N-formyl-methionyl-leucyl-phenylalanine (fMLP) and phorbol myristate acetate (PMA)} was investigated using luminol-amplified chemiluminescence . BHA and the derivative dt-BHA {3,5-di-t-butyl-4-hydroxyanisole} significantly reduced the PMN oxidative burst at concentrations ranging from 5 x 10(-6) to 5 x 10(-5) mol/l for C . albicans stimulation, while for zymosan stimulation, reduction was seen at concentrations ranging from 5 x 10(-6) to 5 x 10(-5) mol/l for BHA, and at concentrations ranging from 5 x 10(-7) to 5 x 10(-5) mol/l for dt-BHA, with dt-BHA being the most active . Another BHA derivative, Bu GAM 1, was active at 5 x 10(-5) mol/l for C . albicans and at 5 x 10(-6) to 5 x 10(-5) mol/l for zymosan . The findings obtained with fMLP and PMA were very similar to those previously reported . ROS release is related to PMN killing activity, but the inhibition of the PMN oxidative burst induced by BHA and BHA derivatives did not significantly modify PMN phagocytosis or killing . It has recently been observed that dt-BHA has a spasmolytic action by inhibiting the influx of Ca(2+) into cells through L-type Ca(2+) channels, which means that a single molecule is capable of counteracting two major steps in the sequence of events triggered by ischemia-reperfusion injury, i.e . free radical release and Ca(2+) overload .

Bioorg Med Chem, 2003 Apr 17, 11(8), 1821 - 7
Antiinflammatory property of 3-aryl-5-(n-propyl)-1,2,4-oxadiazoles and antimicrobial property of 3-aryl-5-(n-propyl)-4,5-dihydro-1,2,4-oxadiazoles: their syntheses and spectroscopic studies; Srivastava RM et al.; The synthesis of six 3-aryl-5-(n-propyl)-4,5dihydro-1,2,4-oxadiazoles 3a-f has been achieved in a facile manner by the reaction of an appropriate arylamidoxime 1a-f with butyraldehyde 2 . Oxidation of 3a-f individually using MnO(2) in CH(2)Cl(2) or sodium hypochlorite in THF/H(2)O furnished 1,2,4-oxadiazoles 4a-f in good to excellent yields . Compounds 4a-f were also evaluated against inflammation . Except 4e, all of them reduced inflammation, however, 4c presented better antiinflammatory activity . A preliminary antimicrobial activity tests of 3a-f showed that these compounds possess activity against some microorganisms . In fact, 3c and 3f have been found to be more effective against Staphylococcus aureus, Mycobacterium smegmatis, and Candida albicans.

J Chromatogr B Analyt Technol Biomed Life Sci, 2003 Apr 5, 787(1), 129 - 48
Analysis of the Candida albicans proteome . II . Protein information technology on the Net (update 2002); Pitarch A et al.; Candida albicans is an important fungal model organism of noteworthy clinical interest in modern medicine . Different initiatives addressing its sequencing and physical mapping have been carried out . The C . albicans genome sequence is currently near to completion at Stanford University, heralding new challenges in proteomic research and functional analyses of its gene products . This review presents an update of the most relevant data resources that are available through the World Wide Web to scientists working in the area of the analysis of the C . albicans proteome . An overview of the current status of the main universal protein sequence databases and specialized data collections for C . albicans is given . Various issues of the single public C . albicans 2D-PAGE database are also described, highlighting the significance of setting up graphical query interface-based databanks to visualize 2D-PAGE images through the Net . Finally, we also emphasize the pressing need to create a "cyber-bioknowledge library" that will integrate all the databases developed at the different levels for the understanding of life processes as well as bioinformatic tools for interpreting this deluge of data generated through the Internet .

J Chromatogr B Analyt Technol Biomed Life Sci, 2003 Apr 5, 787(1), 101 - 28
Analysis of the Candida albicans proteome . I . Strategies and applications; Pitarch A et al.; The alarming incidence of invasive candidiasis, predominantly among the recent expanding immunocompromised population, the appearance of antifungal-drug resistance, and the lack of specific diagnostic tests for it have demanded more impactful research into Candida albicans pathogenicity . Proteomic approaches can provide accurate clues about its biological complexity . Indeed, initial C . albicans proteome analyses have focused on the understanding of dimorphism, host responses, the cell wall, virulence factors and drug resistance, among others . This review aims to briefly outline the technology available for proteomics-based studies, surveying the main proteomic approaches applied to C . albicans research . Prefractionation techniques, two-dimensional gel electrophoresis and mass spectrometry continue to be the backbone of proteomic projects . Emerging strategies for protein separation, quantification and identification may, however, challenge the pivotal position of 2D-PAGE . Regardless of this, since we are now approaching the completion and annotation of C . albicans genome sequencing, systematic characterization of the proteome of this fungal pathogen, although still in its early stages, heralds an exciting expansion of our knowledge in years to come .

Mol Microbiol, 2003 Apr, 48(1), 225 - 35
The Candida albicans Cdr2p ATP-binding cassette (ABC) transporter confers resistance to caspofungin; Schuetzer-Muehlbauer M et al.; Multidrug resistance may pose a serious problem to antifungal therapy . The Candida albicans Cdr2p is one of two ATP-binding cassette (ABC) transporters mediating antifungal resistance in vivo through increased drug efflux . Echinocandins such as caspofungin represent the newest class of antifungals that target cell wall synthesis . We show here by agar plate resistance assays that cross-resistant clinical isolates of C . albicans display high minimal inhibitory concentrations (MICs) to caspofungin when compared with a sensitive ATCC reference strain . Northern analysis and immunoblotting indicate that these isolates also show high levels of CDR1 and CDR2 expression . To determine a possible contribution of Cdr1p or Cdr2p to caspofungin resistance, we have functionally expressed Cdr1p and Cdr2p in appropriate recipient strains of the yeast Saccharomyces cerevisiae . Yeast cells expressing Cdr1p or Cdr2p exhibit cross-resistance to established antifungal drugs such as azoles and terbinafine . However, Cdr2p and, to a much lesser extent, Cdr1p confer caspofungin hyper-resistance when expressed in yeast . Likewise, Cdr2p confers caspofungin resistance when constitutively overexpressed in a drug-sensitive C . albicans strain . We therefore propose that Cdr2p may contribute to clinical candin resistance . Finally, our data suggest that cross-resistance phenotypes of clinical isolates are the consequence of distinct mechanisms that may operate simultaneously.

Mol Microbiol, 2003 Apr, 48(1), 85 - 94
TupA, the Penicillium marneffei Tup1p homologue, represses both yeast and spore development; Todd RB et al.; Fungal pathogenesis is frequently associated with dimorphism - morphological changes between yeast and filamentous forms . Penicillium marneffei, an opportunistic human pathogen, exhibits temperature-dependent dimorphism, with growth at 25 degrees C as filamentous multinucleate hyphae switching at 37 degrees C to uninucleate yeast cells associated with intracellular pathogenesis . The filamentous hyphae also undergo asexual development generating uninucleate spores, the infectious propagules . Both processes require a switch to coupled nuclear and cell division . Homologous regulators, including Tup1p/GROUCHO-related WD40 repeat transcription factors, control dimorphism in Candida albicans and asexual development in Aspergillus nidulans . Unlike these fungi, P . marneffei has both developmental programmes allowing examination of common and programme-specific controls . We show that deletion of tupA, the P . marneffei TUP1 homologue, confers reduced filamentation and inappropriate yeast morphogenesis at 25 degrees C, in stark contrast to constitutive filamentation observed when C . albicans TUP1 is deleted . Deletion of tupA also confers premature brlA-dependent asexual development, unlike reduced asexual development in the corresponding A . nidulans rcoA deletion mutant . Furthermore, the A . nidulans rcoA deletion mutant is self-sterile, and we show that tupA from P . marneffei, which lacks an apparent sexual cycle, complements both the asexual and sexual development phenotypes . Therefore, TupA coordinates cell fate by promoting filamentation and repressing both spore and yeast morphogenetic programmes.

Antimicrob Agents Chemother, 2003 Apr, 47(4), 1220 - 7
Genome-wide expression profile analysis reveals coordinately regulated genes associated with stepwise acquisition of azole resistance in Candida albicans clinical isolates; Rogers PD et al.; Candida albicans is an opportunistic human fungal pathogen and a causative agent of oropharyngeal candidiasis (OPC), the most frequent opportunistic infection among patients with AIDS . Fluconazole and other azole antifungal agents have proven effective in the management of OPC; however, with increased use of these agents treatment failures have occurred . Such failures have been associated with the emergence of azole-resistant strains of C . albicans . In the present study we examined changes in the genome-wide gene expression profile of a series of C . albicans clinical isolates representing the stepwise acquisition of azole resistance . In addition to genes previously associated with azole resistance, we identified many genes whose differential expression was for the first time associated with this phenotype . Furthermore, the expression of these genes was correlated with that of the known resistance genes CDR1, CDR2, and CaMDR1 . Genes coordinately regulated with the up-regulation of CDR1 and CDR2 included the up-regulation of GPX1 and RTA3 and the down-regulation of EBP1 . Genes coordinately regulated with the up-regulation of CaMDR1 included the up-regulation of IFD1, IFD4, IFD5, IFD7, GRP2, DPP1, CRD2, and INO1 and the down-regulation of FET34, OPI3, and IPF1222 . Several of these appeared to be coordinately regulated with both the CDR genes and CaMDR1 . Many of these genes are involved in the oxidative stress response, suggesting that reduced susceptibility to oxidative damage may contribute to azole resistance . Further evaluation of the role these genes and their respective gene products play in azole antifungal resistance is warranted.

Antimicrob Agents Chemother, 2003 Apr, 47(4), 1213 - 9
Genetic basis for differential activities of fluconazole and voriconazole against Candida krusei; Fukuoka T et al.; Invasive infections caused by Candida krusei are a significant concern because this organism is intrinsically resistant to fluconazole . Voriconazole is more active than fluconazole against C . krusei in vitro . One mechanism of fluconazole resistance in C . krusei is diminished sensitivity of the target enzyme, cytochrome P450 sterol 14alpha-demethylase (CYP51), to inhibition by this drug . We investigated the interactions of fluconazole and voriconazole with the CYP51s of C . krusei (ckCYP51) and fluconazole-susceptible Candida albicans (caCYP51) . We found that voriconazole was a more potent inhibitor of both ckCYP51 and caCYP51 in cell extracts than was fluconazole . Also, the ckCYP51 was less sensitive to inhibition by both drugs than was caCYP51 . These results were confirmed by expressing the CYP51 genes from C . krusei and C . albicans in Saccharomyces cerevisiae and determining the susceptibility of the transformants to voriconazole and fluconazole . We constructed homology models of the CYP51s of C . albicans and C . krusei based on the crystal structure of CYP51 from Mycobacterium tuberculosis . These models predicted that voriconazole is a more potent inhibitor of both caCYP51 and ckCYP51 than is fluconazole, because the extra methyl group of voriconazole results in a stronger hydrophobic interaction with the aromatic amino acids in the substrate binding site and more extensive filling of this site . Although there are multiple differences in the predicted amino acid sequence of caCYP51 and ckCYP51, the models of the two enzymes were quite similar and the mechanism for the relative resistance of ckCYP51 to the azoles was not apparent.

Antimicrob Agents Chemother, 2003 Apr, 47(4), 1207 - 12
Drug resistance is not directly affected by mating type locus zygosity in Candida albicans; Pujol C et al.; Recently, evidence was presented that in a collection of fluconazole-resistant strains of Candida albicans there was a much higher proportion of homozygotes for the mating type locus (MTL) than in a collection of fluconazole-sensitive isolates, suggesting the possibility that when cells become MTL homozygous they acquire intrinsic drug resistance . To investigate this possibility, an opposite strategy was employed . First, drug susceptibility was measured in a collection of isolates selected for MTL homozygosity . The majority of these isolates had not been exposed to antifungal drugs . Second, the level of drug susceptibility was compared between spontaneously generated MTL-homozygous progeny and their MTL-heterozygous parent strains which had not been exposed to antifungal drugs . The results demonstrate that naturally occurring MTL-homozygous strains are not intrinsically more drug resistant, supporting the hypotheses that either the higher incidence of MTL homozygosity previously demonstrated among fluconazole-resistant isolates involved associated homozygosity of a drug resistance gene linked to the MTL locus, or that MTL-homozygous strains may be better at developing drug resistance upon exposure to the drug than MTL-heterozygous strains . Furthermore, the results demonstrate that a switch by an MTL-homozygous strain from the white to opaque phenotype, the latter functioning as the facilitator of mating, does not notably alter drug susceptibility.

Antimicrob Agents Chemother, 2003 Apr, 47(4), 1200 - 6
Sublethal injury and resuscitation of Candida albicans after amphotericin B treatment; Liao RS et al.; Amphotericin B treatment was previously shown to inhibit Candida albicans reproduction and reduce the fluorescence of vitality-specific dyes without causing a corresponding increase in the fluorescence of the mortality-specific dyes bis-(1,3-dibutylbarbituric acid)trimethine oxonol and SYBR Green I . In the present study, we have confirmed these results and have shown that the numbers of CFU are reduced by 99.9% by treatment with 0.5 micro g of amphotericin B per ml for 10 h at 35 degrees C . This reduction was not due to fungal cell death . First, the level of reduction of the tetrazolium salt 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-{(phenylamino)carbonyl}-2H-tetrazolium hydroxide increased in the presence of concentrations of amphotericin B that caused greater than 90% reductions in the numbers of CFU . Second, fungal cells treated with amphotericin B at a concentration of 0.5 micro g/ml were resuscitated by further incubation at 22 degrees C for 15 h in the continued presence of amphotericin B . Third, recovery of the ability to replicate was prevented by sequential treatment with 20 micro g of miconazole per ml, which also increased the fluorescence of mortality-specific dyes to near the maximal levels achieved with 0.9 micro g of amphotericin B per ml . Sequential treatment with fluconazole and flucytosine did not increase the levels of staining with the mortality-specific dyes . Itraconazole was less effective than ketoconazole, which was less effective than miconazole . The practice of equating the loss of the capacity of C . albicans to form colonies with fungal cell death may give incorrect results in assays with amphotericin B, and the results of assays with caution with other antifungal agents that are lipophilic or that possess significant postantifungal effects may need to be interpreted.

Antimicrob Agents Chemother, 2003 Apr, 47(4), 1193 - 9
In vivo pharmacodynamics of a new triazole, ravuconazole, in a murine candidiasis model; Andes D et al.; In vivo studies have characterized the pharmacodynamic characteristics of the triazole fluconazole . These investigations demonstrated that the ratio of the area under the concentration-time curve from 0 to 24 h to the MIC (24-h AUC/MIC ratio) is the critical pharmacokinetic/pharmacodynamic (PK/PD) parameter associated with treatment efficacy . Further analysis demonstrated that a fluconazole 24-h AUC/MIC ratio of 20 to 25 was predictive of treatment success in both experimental models and clinical trials . We used a neutropenic murine model of disseminated Candida albicans infection to similarly characterize the time course activity of the new triazole ravuconazole . The PK/PD parameters (percent time above the MIC, AUC/MIC ratio, and peak level in serum/MIC ratio) were correlated with in vivo efficacy, as measured by organism number in kidney cultures after 24 and 72 h of therapy . Ravuconazole kinetics and protein binding were performed in neutropenic infected mice . Peak/dose and AUC/dose values ranged from 0.03 to 0.04 and 0.30 to 0.34, respectively . Serum elimination half-life ranged from 3.9 to 4.8 h . Protein binding was 95.8% . Single-dose postantifungal effect studies demonstrated prolonged suppression of organism regrowth after serum ravuconazole levels had fallen below the MIC . Treatment efficacies with the five dosing intervals studied were similar, supporting the argument for the AUC/MIC ratio as the PK/PD parameter predictive of efficacy . Nonlinear regression analysis also suggested that the AUC/MIC ratio was strongly predictive of treatment outcomes (AUC/MIC ratio, R(2) = 91%; peak/MIC ratio, R(2) = 85%; percent time above the MIC, R(2) = 47 to 65%) . Similar studies were conducted with seven additional C . albicans isolates with various ravuconazole susceptibilities (MIC, 0.016 to 0.12 micro g/ml) to determine if a similar 24-h AUC/MIC ratio was associated with efficacy . The ravuconazole free-drug AUC/MIC ratios were similar for all of the organisms studied (10 to 36; mean +/- SD = 20.3 +/- 8.2; P = 0.43) . These free-drug AUC/MIC ratios are similar to those observed for fluconazole in this model.

Antimicrob Agents Chemother, 2003 Apr, 47(4), 1187 - 92
In vivo pharmacodynamics of HMR 3270, a glucan synthase inhibitor, in a murine candidiasis model; Andes D et al.; In vivo pharmacokinetic/pharmacodynamic characterization for numerous antibacterial compounds has had a significant impact upon optimal dosing regimen design and the development of in vivo susceptibility breakpoints . More recently, similar characterization has been undertaken for antifungal drug classes . Very little is known of these pharmacodynamic relationships for the new echinocandin class of compounds . We utilized a neutropenic murine model of disseminated candidiasis to describe the time course antifungal activity of HMR 3270, a new glucan synthase inhibitor . Single-dose in vivo time kill studies with four 16-fold escalating doses demonstrated concentration-dependent killing when drug levels in serum were more than four times the MIC . Postantifungal effects were dose dependent, ranging from 8 to 80 h duration . Multiple dosing regimen studies utilized six total doses, four dosing intervals, and a treatment duration of 6 days . Shortening the dosing interval from every 144 h (q144h) to q36h resulted in a fourfold rise in the dose necessary to achieve a net fungistatic effect . The peak/MIC ratio most strongly correlated with treatment outcomes (peak/MIC ratio, R(2) = 98%; ratio of the area under the concentration-time curve from 0 to 24 h to the MIC, R(2) = 79%, percentage of time above the MIC, R(2) = 61%) . Studies were also conducted with five additional Candida albicans isolates to determine if a similar peak/MIC ratio was associated with efficacy . In vivo concentration-dependent killing was similarly observed in studies with each of the additional isolates . The peak/MIC ratio necessary to produce efficacy was relatively similar among the strains studied (P = 0.42) . The peak/MIC ratio (mean +/- standard deviation) necessary to achieve a fungistatic effect was 3.72 +/- 1.84, and the ratio necessary to achieve maximal killing was near 10.

Pediatr Transplant, 2003 Apr, 7(2), 93 - 7
Liver transplantation for cholestasis associated with cystic fibrosis in the pediatric population; Molmenti EP et al.; The most common hepatic complications of cystic fibrosis (CF) are steatosis, fibrosis, biliary cirrhosis, atretic gallbladder, cholelithiasis, and sclerosing cholangitis . Cholestatic liver disease is a slow progressive disorder, but will stabilize for many patients . CF patients may suffer from the consequences of their liver disease and without liver transplantation, variceal hemorrhage, malnutrition, or end-stage liver disease can lead to death . Prospective data were collected and reviewed on 311 liver transplants performed in 283 patients at the Children's Medical Center of Dallas between October 1984 and November 2000 . Ten children received an orthotopic liver transplant (OTLX) for end-stage liver disease associated with cystic fibrosis . Pulmonary function tests were obtained preoperatively in all cases . There were nine boys and one girl . Six are currently alive, and four are dead . Both patient and graft survival was 5.75 yr . Among those currently alive, mean patient and graft survival is 7.71 yr (range 0.10-12.62 yr) . Mean patient and graft survival of those who died was 2.35 yr (range 0.78-5.33 yr) . No survivor required re-transplantation and currently, all have normal serum aminotransferase values . Chronic sinusitis was not a significant pre- or post-transplant morbidity, although systematic radiographic evaluation of the sinuses did not occur . Pulmonary deaths occurred in three patients from pulmonary hemorrhage, pulmonary infection with Aspergillus and Candida glabrata, and acute bronchopneumonia associated with polymicrobial sepsis because of Pseudomonas, Klebsiella, and Candida albicans 1.44, 0.78, and 1.83 yr, respectively, after transplantation . The fourth death was associated with chronic rejection, and occurred 5.33 yr after transplantation . All non-survivors were below the 5th percentile for height and weight at the time of liver transplantation . Mean age at transplantation was 9.72 yr (range 1.23-19.09, median 9.61) . Survivors were transplanted at a younger age than non-survivors (mean of 9.21 yr vs . 10.66 yr), and had shorter waiting times from diagnosis of end-stage liver disease to transplantation (6.87 months vs . 13.83 months) . Eighty percentage (n = 8) of patients had pretransplant variceal bleeds (83% of survivors, 75% of non-survivors) . While all non-survivors had a history of meconium ileus and preoperative need of pancreatic enzymes, only 67% of those alive experienced these complications . Preoperative forced vital capacity FVC was 103% for survivors and 95% for non-survivors . The corresponding numbers for forced expiratory flow (FEF) 25-75 were 74-84% respectively . Preoperative Aspergillus was identified in 30% of patients (n = 3) . Two of these patients are alive . Cystic fibrosis constitutes an indication for 3.5% of pediatric liver transplants . Evaluation and transplantation for end-stage liver disease associated with cystic fibrosis should be undertaken at an early age . Most deaths were associated with pulmonary/septic events, and occurred less than 2 yr after OLTX . Those children who did not survive had poor growth and nutrition, prolonged waiting times prior to transplantation, were transplanted at an older age, and had a higher incidence of pancreatic insufficiency and meconium ileus . The presence of Aspergillus in the sputum does not constitute a contraindication for OLTX.

Mycopathologia, 2002, 155(4), 195 - 201
The fungicidal activity of novel nanoemulsion (X8W60PC) against clinically important yeast and filamentous fungi; Myc A et al.; Surfactant nanoemulsions are water in oil preparations that proved to have a broad spectrum biocidal activity against a variety of microorganisms including Gram-positive and Gram-negative bacteria, spores and enveloped viruses . These preparations are non-toxic to the skin, mucous membrane and gastrointestinal tissues at biocidal concentrations . In this study, 0.1% of the nanoemulsion designated X8W60PC has shown fungicidal activity against yeast including Candida albicans and C . tropicalis in 15 minutes . C . tropicalis was more sensitive than C . albicans, which required a longer time or a higher concentration of the nanoemulsion to achieve killing . Neutral to slightly alkaline pH was more effective in killing the yeast cells than acidic pH . Using the minimum inhibitory concentration assay, 0.08% of the nanoemulsion was inhibitory to C . albicans, and parapsilosis and filamentous fungi including Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, Aspergillus fumigatus and Fusarium oxysporum . None of the individual ingredients was as effective a fungicidal as the nanoemulsion at equivalent concentration . This shows that the nanoemulsion structure is an important factor in the anti-fungal activity . The X8W60PC has great potential as a topical anti-fungal agent and further investigation into the mechanism of fungicidal action is warranted.

Int Rev Immunol, 2002 Nov-Dec, 21(6), 515 - 48
The protective immune response against vaginal candidiasis: lessons learned from clinical studies and animal models; Fidel PL Jr; Recurrent vulvovaginal candidiasis (RVVC) is a significant problem in women of childbearing ages and is caused by Candida albicans, a commensal organism of the intestinal and reproductive tracts . As a result of this commensalism, most healthy individuals have demonstrable Candida-specific adaptive immunity that is considered protective . In women with RVVC, a deficiency/dysfunction of this protective immunity is postulated to affect susceptibility to infection . Although cell-mediated immunity (CMI) is considered important for protection against mucosal candidal infections, little is understood about specific host defenses that are important at the vaginal mucosa . Studies to date suggest that a compartmentalized local, rather than systemic, immunity is important for defense against vaginitis . This review will summarize the current state of knowledge regarding protective host defense mechanisms against vaginal C . albicans infections both from clinical studies and animal models . From these data, hypotheses are presented for what host defense mechanisms appear important for resistance/susceptibility to vaginal infection.

Phytochemistry, 2003 Apr, 62(8), 1251 - 5
Muscanone: a 3-O-(1", 8", 14"-trimethylhexadecanyl)naringenin from Commiphora wightii; Fatope MO et al.; A new antifungal flavanone, muscanone (1), was isolated along with known naringenin (2) from Commiphora wightii (Arn.) Bhandari (Burseraceae) by directing the fractionation of an EtOH extract of the air-dried trunk of C . wightii with microbial sensitivity assay . The structures of 1 and 2 were determined from EIMS, HREIMS, DEPT, 1H-1H COSY, HSQC and HMBC spectral data . Muscanone (1) was identified as 3-O-(1", 8",14"-trimethylhexadecanyl)naringenin and was found to be active against Candida albicans . The isolation, structure elucidation, NMR spectral assignments, and bioactivities of 1 and 2 are reported .

Infect Dis Obstet Gynecol, 2002, 10(4), 193 - 202
Case-control study of vulvar vestibulitis risk associated with genital infections; Smith EM et al.; OBJECTIVE: To evaluate the risk of vulvar vestibulitis syndrome (VVS) associated with genital infections in a case-control study . METHODS: Diagnosed cases with VVS (n = 69) and age-frequency-matched healthy controls (n = 65) were enrolled from gynecology clinics in a university medical hospital during 1999 . They were compared for potential risk factors and symptoms of disease . RESULTS: VVS cases had a significantly higher risk of physician-reported bacterial vaginosis (BV) (odds ratio, OR = 9.4), Candida albicans (OR = 5.7), pelvic inflammatory disease (PID) (OR = 11.2), trichomoniasis (OR = 20.6), and vulvar dysplasia (OR = l5.7) but no risk associated with human papillomavirus (HPV), ASCUS, cervical dysplasia, genital warts, chlamydia, genital herpes or gonorrhea . Genital symptoms reported significantly more often with VVS included vulvar burning (91 vs . 12%), dyspareunia (81 vs . 15%), vulvar itching (68 vs . 23%) and dysuria (54 vs . 19%) (p < 0.0001) . CONCLUSION: A history of genital infections is associated with an increased risk of VVS . Long-term follow-up case-control studies are needed to elucidate etiologic mechanisms, methods for prevention and effective treatment.

J Immunol, 2003 Apr 1, 170(7), 3843 - 9
Unique regulation of CCL18 production by maturing dendritic cells; Vulcano M et al.; Dendritic cells (DC) orchestrate the trafficking of lymphocytes by secreting chemokines with different specificity and function . Chemokines are produced at higher levels by mature DC . This study shows that CCL18 is one of the most abundant chemokines produced by immature DC . In contrast to all other chemokines investigated to date, CCL18 was selectively down-regulated during the maturation process induced by LPS, TNF, CD40 ligand, Staphylococcus aureus Cowan I, Candida albicans, and influenza virus . IL-10 and vitamin D(3), two known inhibitors of DC differentiation and function, strongly promoted CCL18 secretion, whereas IFN-gamma, a costimulator of DC function, inhibited its production . IL-10 also induced CCL18 secretion in blood myeloid DC . No CCL18 secretion was observed in blood plasmacytoid DC . The opposite pattern of regulation was observed for CCL20, a prototypic inflammatory chemokine . CCL18 was found to be a chemotactic factor for immature DC . Therefore, CCL18 may act as a chemotactic signal that promotes the colocalization of immature DC with naive T lymphocytes in an IL-10-dominated environment with the consequent generation of T regulatory cells . These characteristics suggest that CCL18 may be part of an inhibitory pathway devoted to limiting the generation of specific immune responses at peripheral sites.

Eur J Immunol, 2003 Feb, 33(2), 532 - 8
The C-type lectin DC-SIGN (CD209) is an antigen-uptake receptor for Candida albicans on dendritic cells; Cambi A et al.; Dendritic cells (DC) that express the type II C-type lectin DC-SIGN (CD209) are located in the submucosa of tissues, where they mediate HIV-1 entry . Interestingly, the pathogen Candida albicans, the major cause of hospital-acquired fungal infections, penetrates at similar submucosal sites . Here we demonstrate that DC-SIGN is able to bind C . albicans both in DC-SIGN-transfected cell lines and in human monocyte-derived DC . The binding was shown to be time- as well as concentration-dependent, and live as well as heat-inactivated C . albicans were bound to the same extent . Moreover, in immature DC, DC-SIGN was able to internalize C . albicans in specific DC-SIGN-enriched vesicles, distinct from those containing the mannose receptor, the other known C . albicans receptor expressed by DC . Together, these results demonstrate that DC-SIGN is an exquisite pathogen-uptake receptor that captures not only viruses but also fungi.

J Investig Med, 2003 Mar, 51(2), 95 - 103
Silica, hyaluronate, and alveolar macrophage functional differentiation; Bodo M et al.; BACKGROUND: Silicosis is mediated by macrophages, their soluble mediators, and extracellular matrix molecules . In this study, we investigated the effects of silica and/or hyaluronate (HA) on several alveolar macrophage responses . METHODS: We evaluated glycosaminoglycan (GAG) production by radiolabeled precursors, nitric oxide (NO) release by its oxidation product, phagocytic activity by Candida albicans internalization, and the secretion of two fibrogenic cytokines, tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta, by specific assays . RESULTS: Silica significantly reduced GAG secretion, particularly HA secretion . Alone, it decreased Candida uptake; associated with HA, it enhanced the reduction . Silica and Candida reduced NO release, which was not significantly affected when silica- or Candida-exposed cells were also treated with HA . TNF-alpha and TGF-beta activities were stimulated by silica but reduced by HA . CONCLUSIONS: The results suggest that silica and HA modify alveolar macrophage functional differentiation . Silica- and HA-induced modifications of the microenvironment could determine whether the response proceeds toward healing and repair or toward lung chronic pathology.

Arch Microbiol, 2003 Apr, 179(4), 295 - 300 Epub 2003 Mar 15.
Disruption of mitochondrial function in Candida albicans leads to reduced cellular ergosterol levels and elevated growth in the presence of amphotericin B; Geraghty P et al.; A respiratory-deficient mutant of Candida albicans MEN was generated by culturing cells in medium supplemented with ethidium bromide at 37 degrees C for 5 days . The respiratory-deficient mutant (C . albicans MMU11) was incapable of growth on glycerol, had a reduced oxygen uptake rate and demonstrated an altered mitochondrial cytochrome profile . Respiratory-competent cybrids were formed by mitochondrial transfer following fusion of protoplasts with those of C . albicans ATCC 44990 . Mutant MMU11 possessed lower levels of ergosterol than the parental isolates and the cybrids, and demonstrated a small but statistically significant increase in tolerance to amphotericin B . The results demonstrated that disruption of mitochondrial function in C . albicans increases the tolerance to amphotericin B, possibly mediated by a reduction in cellular ergosterol content.

Kansenshogaku Zasshi, 2003 Jan, 77(1), 29 - 33
Effect of amphotericin B dilution with various beverages on the survival of Candida albicans cells; Ohshima T et al.; Amphotericin B (AMPH) has been generally used for prophylaxis or treatment of specific fungal diseases in immunocompromised patients . However, because it is difficult for children to ingest, mainly because of its bitter taste, it is often diluted with soft drinks . We therefore investigated the effect of dilution of AMPH with various beverages on its antifungal activity in vitro . Candida albicans cells were exposed for 30 min to AMPH diluted twofold with each of six commercially available beverages or distilled water, and percent survival was determined . The results showed 60% survival in the dilution with distilled water and higher survival when diluted with Yakult (136%; p < 0.01), orange juice (104%; p < 0.01), and coffee-milk (92%; p < 0.01) . By contrast, lower survival was obtained when diluted with gum-syrup (54%), sweet cider (76%), and shaved-ice syrup (52%) with no significant differences from distilled water (60%), suggesting that these three beverages may be useful for diluting AMPH . The results of this study are a warning to medical workers that some methods of making AMPH more palatable considerably decrease its antifungal activity and may have a negative effect on host defenses against infectious diseases.

Biochim Biophys Acta, 2003 Mar 21, 1646(1-2), 184 - 95
Kinetic and mechanistic analysis of the association and dissociation of inhibitors interacting with secreted aspartic acid proteases 1 and 2 from Candida albicans; Backman D et al.; In order to elucidate the characteristics of different aspartic proteases (Sap) secreted by Candida albicans, the kinetics of the interaction (k(on), k(off)) between Sap1 and Sap2 with acetyl-pepstatin and pepstatin A was determined at different pH by biosensor technology . The enzymes were biotinylated and coupled to a streptavidin-coated sensor chip, whereupon acetyl-pepstatin or pepstatin A was injected and the interaction was measured in real time . Sap2 showed a faster k(on) and a higher affinity for acetyl-pepstatin than Sap1, regardless of pH . The values for both k(on) and k(off) decreased with increased pH from 3.8 to 5.0, except for the k(off) for Sap1, which was only influenced by the pH change from 3.8 to 4.4 . Binding of acetyl-pepstatin to Sap1 or Sap2 obviously proceeds by a different mechanism than dissociation of the inhibitor . Association appears to be coupled to protonation of a catalytic aspartic acid residue, consistent with reduced k(on) values at higher pH . In contrast, the stability of the complex is reduced at lower pH due to reduced hydrogen bonding capacity of aspartic acid residues acting as hydrogen bond acceptors . Differences in the number and distribution of charged nonactive site residues in Sap1 and Sap2 evidently result in different electrostatic properties of the binding sites, primarily influencing the association step.

Microbiol Immunol, 2003, 47(1), 37 - 43
Lactoferrin feeding augments peritoneal macrophage activities in mice intraperitoneally injected with inactivated Candida albicans; Wakabayashi H et al.; Oral administration of lactoferrin (LF), an innate-defense protein present in exocrine secretions such as milk and in neutrophils, is reported to improve host-protection against infections with microorganisms including pathogenic fungi, possibly due to an immunomodulatory effect . This study aimed to evaluate the effect of bovine LF feeding on peritoneal macrophage activities in mice intraperitoneally injected with inactivated Candida albicans . Time course analysis during the 14 days following Candida-priming revealed that LF administration slightly increased the number of peritoneal exudate cells, and significantly enhanced the production of superoxide anion (O2(-)) and nitric oxide (NO) by peritoneal macrophages at day 7 . LF administration facilitated NO production and Candida hyphal-growth inhibition by macrophages derived from Candida-primed mice but not non-primed mice, suggesting that the action of LF is dependent on the immune status of the host . LF administration altered the kinetics of cytokines in the peritoneal lavage fluid of Candida-primed mice . Enhancement of cytokine levels by LF was observed for IL-12 at day 5 and IFN-gamma at day 9, but not for TNF-alpha or IL-10 . In conclusion, LF feeding augmented the activities of macrophages in a manner dependent on Candida-priming and these effects may be related to enhanced cytokine levels.

Indian J Exp Biol, 2002 Mar, 40(3), 314 - 8
Fungistatic activity of Semecarpus anacardium Linn . f nut extract; Sharma K et al.; Alcoholic extract of dry nuts of S . anacardium showed dose dependent antifungal activity in vitro against Aspergillus fumigatus and Candida albicans . At 400 mg/ml concentration, growth of both the fungi was inhibited and considerable reduction in size of cells and hyphae was observed . Sporulation also decreased.

Microbiology, 2003 Mar, 149(Pt 3), 579 - 88
Haemin uptake and use as an iron source by Candida albicans: role of CaHMX1-encoded haem oxygenase; Santos R et al.; Candida albicans, unlike Saccharomyces cerevisiae, was able to use extracellular haemin as an iron source . Haemin uptake kinetics by C . albicans cells showed two phases: a rapid phase of haemin binding (with a K(d) of about 0.2 microM) followed by a slower uptake phase . Both phases were strongly induced in iron-deficient cells compared to iron-rich cells . Haemin uptake did not depend on the previously characterized reductive iron uptake system and siderophore uptake system . CaHMX1, encoding a putative haem oxygenase, was shown to be required for iron assimilation from haemin . A double DeltaCahmx1 mutant was constructed . This mutant could not grow with haemin as the sole iron source, although haemin uptake was not affected . The three different iron uptake systems (reductive, siderophore and haemin) were regulated independently and in a complex manner . CaHMX1 expression was induced by iron deprivation, by haemin and by a shift of temperature from 30 to 37 degrees C . CaHMX1 expression was strongly deregulated in a Deltaefg1 mutant but not in a Deltatup1 mutant . C . albicans colonies forming on agar plates with haemin as the sole iron source showed a very unusual morphology . Colonies were made up of tubular structures that were organized into a complex network . The effect of haemin on filamentation was increased in the double DeltaCahmx1 mutant . This study provides the first experimental evidence that haem oxygenase is required for iron assimilation from haem by a pathogenic fungus.

Med Dosw Mikrobiol, 2002, 54(3), 273 - 9
{Presence of fungi in stool of children}; Pawlik B et al.; A total of 258 children were tested for the presence of fungi in stool . One group consisted of 148 children with non-specific gastrointestinal tract disorders while the other was a group of 110 asthmatics . A quantitative method of enzymatic and mechanical homogenisation was used . The findings were divided into three ranges as follows: < 10(3), 10(3)-10(5), > 10(5) fungal cells in one gram of stool . The number of > 10(5) fungal cells in one gram of stool was considered as pathogenic and requiring treatment . Such a number of fungi in stool was found in 48.1% of children in the first group and in 35.9% in the second one . However, the percentage of fungal presence was higher in the group of asthmatics (83.6% vs . 70.3%) . Candida albicans considerably outnumbered the remaining fungal species in the isolates . It was found out that other than C . albicans Candida species were more resistant to the antifungals.

J Pharm Pharmacol, 2003 Feb, 55(2), 179 - 84
Erythromycin, an inhibitor of mitoribosomal protein biosynthesis, alters the amphotericin B susceptibility of Candida albicans; Geraghty P et al.; Exposure of the yeast Candida albicans to the macrolide antibiotic erythromycin (C(37)H(67)NO(13)) results in elevated tolerance to the polyene antifungal amphotericin B . Erythromycin displays no fungistatic activity against C . albicans but inhibits the synthesis of cytochromes, particularly cytochrome aa(3) . Consequently there is a reduction in aerobic respiration by up to 90% when cells are exposed to 10 mg mL(-1) erythromycin . Cellular ergosterol levels are also severely reduced . Erythromycin inhibits protein biosynthesis in ribosomes (mitoribosomes) located within the mitochondrion of the yeast cell, which results in a disruption of cytochrome biosynthesis with an adverse effect on respiration . The synthesis of ergosterol is oxygen dependent and consequently ergosterol levels are depleted in erythromycin-treated C . albicans . Ergosterol is the target for amphotericin B and since there is less of this sterol in erythromycin-treated cells, there is an increase in tolerance of the antifungal agent . Our work indicates that co-administration of erythromycin and amphotericin B to control bacterial and fungal infections, respectively, may inadvertently lead to an elevation in the tolerance of C . albicans for this antifungal agent.

Shock, 2003 Mar, 19(3), 257 - 62
Hypoxia and extraintestinal dissemination of Candida albicans yeast forms; Kim AS et al.; Candida albicans is a pleomorphic fungus with budding yeast and filamentous forms, and is a frequent cause of complicating infections in patients who are postsurgical, in shock, and have trauma . Many cases of systemic candidiasis are thought to orginate from the intestine, but it is unclear if the filament or the yeast is the more invasive form . Because C . albicans is relatively noninvasive and because mesenteric ischemia is thought to facilitate extraintestinal microbial dissemination, wild-type C . albicans CAF2 and mutant HLC54 (defective in filament formation) were orally inoculated into antibiotic-treated mice that were housed exclusively in room air, or were intermittently exposed to 10% oxygen for 1-h intervals . Both strains of C . albicans colonized the cecum in similar numbers (approximately 10(6.7)/g) . C . albicans translocation to the draining mesenteric lymph nodes was not detected in mice inoculated with CAF2 (normoxic or hypoxic) or in normoxic mice inoculated with HLC54, but was detected in 33% (P < 0.01) of hypoxic mice inoculated with HLC54 . Using Caco-2 and HT-29 enterocytes cultivated on plastic dishes and pretreated for 48 h in 10% oxygen, adherence of C . albicans HLC54 was decreased compared with wild-type CAF2, and hypoxia had no noticeable effect on adherence of either CAF2 or HLC54 . Using enterocytes cultivated on permeable 8-microm filters, transepithelial migration of C . albicans CAF2 and HLC54 appeared similar . Thus, C . albicans HLC54 (defective in filament formation) was more invasive in hypoxic mice compared with wild-type CAF2, and host factors (e.g., mesenteric ischemia) rather than an innate ability to interact with enterocytes might play a more important role in extraintestinal dissemination of C . albicans yeast forms.

Folia Microbiol (Praha), 2002, 47(6), 732 - 6
Detection of anti-Candida antibodies by the indirect immunofluorescence assay in patients with cancer in the orofacial region; Dorko E et al.; An indirect immunofluorescence assay was performed to detect antibodies to Candida albicans blastospores and germ tubes . Serum specimens were obtained from 82 patients with neoplastic diseases in the orofacial region and thrush of the oral mucosa . C . albicans was identified in the oral cavity of 63 patients investigated but serum anti-Candida antibodies were detected in only 23 of them . Serological examination showed that titers of antibodies to C . albicans blastospores ranged from 1:20 to 1:1280 . High titers from 1:640 to 1:1280 were detected in patients without antibiotic, cytostatic, or radiotherapeutic treatment . The titers of antibodies to C . albicans germ tubes ranged from 1:20 to 1:640 . Our results indicate that titers of antibodies to the C . albicans germ tubes were lower and were detected in a smaller number of patients.

Folia Microbiol (Praha), 2002, 47(6), 727 - 31
The frequency of Candida species in onychomycosis; Dorko E et al.; Mycological investigation of 108 nail specimens taken from a total of 41 patients examined over three years included direct microscopy and repeated cultures . A higher incidence of onychomycosis of the fingernails (75%) was observed in women while afflictions of the toenails (71%) prevailed in men . The highest prevalence of onychomycosis was found in patients between 50 and 70 years of age . Candida albicans was the dominant organism causing onychomycosis (prevalence rate 60.9%), followed by C . parapsilosis (19.6%), C . tropicalis (9.8), C . krusei (4.9), C . guilliermondii and C . zeylanoides (2.4% each).

FEMS Immunol Med Microbiol, 2003 Mar 20, 35(2), 159 - 64
Impaired neutrophil function in patients with pulmonary tuberculosis and its normalization in those undergoing specific treatment, except the HIV-coinfected cases; Fiorenza G et al.; Our study investigated whether the respiratory burst (RB) of polymorphonuclear neutrophils from tuberculosis (TB) patients was related with the disease severity or treatment, as well as the circulating levels of TNF-alpha . The sample comprised 57 patients with moderate (n=21) or advanced disease (n=36, 13 of them with HIV coinfection, TB-HIV) and 12 controls . Patients were newly diagnosed (n=27) or under treatment (moderate=14, advanced=10, TB-HIV=6) . Cytometric analysis showed that untreated patients had a depressed RB in response to Candida albicans, being more pronounced in the advanced group and nearly absent in TB-HIV cases . A recovered RB was observed in treated patients, except for the TB-HIV cases that continued to show a poor response . TNF-alpha serum levels were increased in untreated patients, mostly in the advanced and TB-HIV groups, and showed an inverse and significant correlation with the RB . Disease severity and anti-TB therapy exerted negative and positive influences on the reactive oxygen intermediates production, respectively.

FEMS Immunol Med Microbiol, 2003 Mar 20, 35(2), 147 - 52
Flow cytometric measurements of neutrophil functions: the dependence on the stimulus to cell ratio; Anding K et al.; Phagocytosis and antimicrobial killing of neutrophils has been quantitatively determined as a function of the stimulus (Candida albicans) to cell ratio R using two donor collectives containing a total of 115 blood samples . Analysis of the collectives in two different laboratories according to the same flow cytometric protocol for simultaneous measurement of neutrophil functions did not produce statistically significant differences . The number of phagocytosing leukocytes as well as that of killed fungi per leukocyte depends strongly on R . While each phagocytosing neutrophil kills one fungus at low values of R, each neutrophil kills on average 2.5 fungi for large R.

Med Mycol, 2003 Feb, 41(1), 53 - 8
Isolation and purification of chiamydospores of Candida albicans; Fabry W et al.; Two methods were investigated for their efficiency in isolating and purifying chlamydospores of Candida albicans . Chlamydospores were disconnected from pseudomycelial cells either enzymatically using beta-glucuronidase or mechanically by ultrasonic treatment . Free chlamydospores were separated from other cell material by sucrose gradient centrifugation . The resulting preparations were inspected by light-microscopy and electron-microscopy . Both methods yielded preparations with a level of over 90% chlamydospore cells . Ultrasonic treatment caused little change to the ultrastructure of the chlamydospores, whereas the enzyme treatment profoundly affected the cell wall . It is concluded that ultrasonic treatment is an efficient method for obtaining pure preparations of chlamydospores.

Med Mycol, 2003 Feb, 41(1), 43 - 52
Isolation and characterization of an avirulent Candida albicans yeast monomorphic mutant; Iranzo M et al.; Mutagenesis of Candida albicans strain ATCC 26555 with N-methyl-nitro-N-nitrosoguanidine followed by plating on solid yeast nitrogen base-N-acetylglucosamine medium at 37 degrees C yielded colony morphology variants that were characterized as forming smooth colonies, in contrast to the rough colonies formed by the parental strain . One yeast monomorphic mutant, CAL4, was studied in detail . Strain CAL4 is defective in filamentous growth, unable to form hyphae or pseudohyphae in vivo and in vitro . These filamentous structures are not elicited by commonly used external stimuli such as serum . The mutant had no obvious alterations in its mannan, glucan or chitin content . The total quantity of non-covalently linked wall proteins was reduced in the mutant strain, but the electrophoretic pattern shown by these proteins was identical to that of proteins from the parental strain . CAL4 showed major differences from the parental strain in its formation of covalently linked wall proteins . An important aspect of these differences lay in the practical absence of proteins recognized by two monoclonal antibodies, 1B12 and 3H8, which are considered valuable tools in the diagnosis of candidiasis in part because they normally react strongly with all strains . The C . albicans mutant, blocked in yeast-mycelium transition, was avirulent in a mouse model, although it was able to grow in animal tissues.

Med Mycol, 2003 Feb, 41(1), 21 - 30
Candida albicans cell wall antigens for serological diagnosis of candidemia; Kondori N et al.; Serological tests for diagnosis of disseminated fungal infections in the immunocompromised host are used with varying results . In the present study, the relative ability of antibodies to specifically recognize Candida albicans cell wall components was evaluated in order to find antigenic markers for serological diagnosis of candidemia . Native C . albicans cell wall fragments (CW), periodate- (CWIO4) and proteinase-K- (CWP) treated CW, a mildly extracted phosphopeptidomannan (PPM), and beta(1-3)(1-6)-glucan were used as antigens in ELISA with sera from rabbits immunized with C . albicans (n = 10), patients with culture proven candidemia (n = 8) and healthy individuals (n = 8) . The antibody response in rabbits consisted predominantly of anti-PPM antibodies, a finding that was substantiated by inhibition-ELISA . Consistently, periodate treatment (CW104) destroyed a major proportion of the antigenic epitopes . Low rabbit antibody levels were found against glucan, the major Candida cell wall component . These results supported the conclusion that glucan is localized mainly in the inner part of the C . albicans cell wall . In contrast to rabbits' serum IgG antibody response against PPM, which was at least tenfold higher than that raised against CW, patients with candidemia had similar IgG antibody levels against both antigens . These levels were significantly higher than those seen in healthy controls (CW, P = 0.0005 and PPM, P < 0.0001) . Although the human anti-glucan and anti-CWIO4 IgG antibody levels were low overall, they were nonetheless significantly increased in the patient group (P = 0.0159 for antiglucan and P = 0.0491 for anti-CWIO4) . In addition, a correlation was noticed between levels of these antibodies . No significant differences were found between patients and controls for IgM antibodies when CW, CWIO4, PPM and Glu were used as antigens . In conclusion, IgG antibodies to PPM and native cell wall fragments (CW) were highly discriminatory for recognition of candidemia and these antigens are thus promising candidates for use in serodiagnosis.

Med Mycol, 2003 Feb, 41(1), 7 - 14
Analyses of phagocytosis, evoked oxidative burst, and killing of black yeasts by human neutrophils: a tool for estimating their pathogenicity?
Peltroche-Llacsahuanga H, Schnitzler N, Jentsch S, Platz A, De Hoog S, Schweizer KG, Haase G.
The pathogenicity of several dematiaceous yeasts that have, to date, rarely been isolated in humans remains unclear . Because professional phagocytes are prominent in lesions caused by dematiaceous fungi, we address this issue by comparing phagocytosis, evoked oxidative burst and killing by human neutrophils of different black yeasts in vitro . Whereas phagocytosis of all black yeasts tested and evoked oxidative burst yielded comparable results, in contrast, the degree of killing differed significantly after 5 h . Thereby, two groups could be identified; one in which strains are killed at high rates, for example, Hortaea werneckii (81 +/- 11.6%), Exophiala castellanii (96 +/- 8.6%), Phaeoannellomyces elegans (93 +/- 9.7%), Phaeococcomyces exophialae (87 +/- 8.7%), and the other in which strains are killed to a lesser degree, for example, Exophiala dermatitidis (ATCC 34100) (61 +/- 9.5%), E . dermatitidis (CBS 207.35) (66 +/- 7.5%), E . jeanselmei (50 +/- 10.5%), E . mesophila (63 +/- 11.6%), E . bergeri (63 +/- 9.1%), and E . spinifera (57 +/- 9.6%) . Non-pigmented yeasts were killed at levels comparable with those at which the white mutant strain of E . dermatitidis (ATCC 44504) was killed (95 +/- 7.5%); the yeast strains tested were Candida albicans (DSM 11943) (95 +/- 4.0% killing) and Saccharomyces cerevisiae (DSM 1333) (95 +/- 10.3%) . Comparison of killing rates with the observed pathogenicity of the melanized species suggests that low killing rates might indicate or even predict a high degree of invasiveness . Although previous experiments revealed that melanization conferred killing resistance on E . dermatitidis, the differences in killing rates of other dematious fungi suggest that melanization of the cell wall is in itself insufficient to confer virulence.

Med Mycol, 2003 Feb, 41(1), 1 - 6
Reflections on the question: what does molecular mycology have to do with the clinician treating the patient?
Odds FC.
After a talk on regulation of gene transcription in Candida albicans, a clinical mycologist was heard to ask: "What difference does all that make to the lady dying of disseminated Candida infection in her hospital bed?" The rapid expansion of research in fungal diseases is widening the communication gap between individuals with responsibility for patient care and those who study pathogenic fungi at the level of molecular biology . DNA-based technologies have produced real advances for patient care by delivering superior methods for fungus identification and strain typing that will soon find a routine place in patient management . Molecular research into the fine detail of the host-pathogen interplay in fungal disease has also made great advances, though the spin-offs to benefit the clinician are not yet obvious . Detection of fungal DNA as a non-culture diagnostic method still requires considerable refinement before it can be regarded as a routinely useful approach, and genomic-based strategies for discovery of novel antifungal drugs from molecular targets have so far produced no agents that have entered development . It is inevitable that, in time, several aspects of molecular mycology research will become important basic knowledge for the clinician who is treating the patient . Therefore, clinicians and bench scientists with specialist interest in mycoses need to retain a reasonable level of mutual comprehension and respect of each other's work rather than assuming their professional paths are divergent.

Proteomics, 2003 Mar, 3(3), 325 - 36
Drug induced proteome changes in Candida albicans: comparison of the effect of beta(1,3) glucan synthase inhibitors and two triazoles, fluconazole and itraconazole; Bruneau JM et al.; The dimorphic fungus Candida albicans is an opportunistic human pathogen . Candidiasis is usually treated with azole antifungal agents . However clinical treatments may fail due to the appearance of resistance to this class of antifungal agents in Candida . Echinocandin derivatives are an alternative for the treatment of these fungal infections and are active against azole resistant isolates of C . albicans . Azoles inhibit the lanosterol 14 alpha demethylase which is a key enzyme in the synthesis of ergosterol . In contrast, the echinocandin class of antibiotics inhibit noncompetitively beta-(1,3)-D-glucan synthesis in vitro . We have investigated the impact of mulundocandin on the proteome of C . albicans and compared it to those of a mulundocandin derivative, as well as to two azoles of different structure, fluconazole and itraconazole . The changes in gene expression underlying the antifungal responses were analyzed by comparative 2-D PAGE . Dose dependant responses were kinetically studied on C . albicans grown at 25 degrees C (yeast form) in synthetic dextrose medium . This study shows that antifungals with a common mechanism of action lead to comparable effects at the proteome level and that a proteomics approach can be used to distinguish different antifungals, with the promise to become a useful tool to study drugs of unknown mechanism of action.

Planta Med, 2003 Feb, 69(2), 158 - 61
Composition and antimicrobial activity of the essential oil of Artemisia absinthium from Croatia and France; Juteau F et al.; The essential oils obtained by steam distillation from the aerial parts of two populations of Artemisia absinthium, from France and from Croatia, were analyzed by GC and GC-MS . The oils of A . absinthium of French origin contain (Z)-epoxyocimene and chrysanthenyl acetate as major components while the oils of Croatian A . absinthium contain mainly (Z)-epoxyocimene and beta-thujone . Analysis of oils before and after anthesis showed some quantitative differences . Analysis of separated leaves and flowering heads showed only few differences among these organs . As they contain no thujone, antimicrobial screening was performed on samples of French origin and showed that A . absinthium oil inhibited the growth of both tested yeasts (Candida albicans and Saccharomyces cerevisiae var . chevalieri).

Microbiology, 2003 Feb, 149(Pt 2), 333 - 9
Increased resistance in BALB/c mice to reinfection with Candida albicans is due to immunoneutralization of a virulence-associated immunomodulatory protein; Tavares D et al.; Here, it is shown that immunoneutralization of p43, a virulence-associated immunomodulatory protein secreted by Candida albicans, is responsible for immunoprotection against candidiasis after spontaneous healing of mice inoculated with 10(6) C . albicans blastoconidia . p43 is produced by the pathogenic Candida blastoconidia, and neither immunoprotection nor immunoneutralization can be elicited by priming the mice with attenuated or heat-killed C . albicans blastoconidia . The immunoprotection against systemic candidiasis was positively correlated with (i) . serum levels of antibodies against p43 and (ii) . a high ratio between antibodies against p43 and antibodies against C . albicans structural antigens . Immunoprotection against candidiasis can be induced in mice primed with heat-killed C . albicans, after treatment of the animals with anti-p43 antibodies . The data described here provide a biological explanation for active immunoprotection achieved after spontaneous healing of infectious diseases, namely in candidiasis.

J Clin Microbiol, 2003 Mar, 41(3), 1316 - 21
Stability of allelic frequencies and distributions of Candida albicans microsatellite loci from U.S . population-based surveillance isolates; Lott TJ et al.; Allelic distributions and frequencies of five Candida albicans microsatellite loci have been determined for strains isolated from the bloodstream and obtained through active population-based surveillance in two U.S . metropolitan areas between 1998 and 2000 . These data were compared to data for isolates obtained from two other U.S . regions in 1992 to 1993 . In a majority of pairwise combinations between sites, no evidence was seen for shifts in microsatellite allelic frequencies . One to three alleles were highly predominant and correlated with major genotypes . These data both support the concepts of allelic stability and genetic equilibria and suggest that, in the United States, strains of C . albicans isolated from the bloodstream may form a defined, genetically homogeneous population across geographical distance and time.

J Clin Microbiol, 2003 Mar, 41(3), 1259 - 62
Casein agar: a useful medium for differentiating Candida dubliniensis from Candida albicans; Mosca CO et al.; Production of chlamydospores on casein agar at 24 degrees C for 48 h provides a simple means for differentiating Candida dubliniensis from Candida albicans based on chlamydospore production . Of 109 C . dubliniensis isolates tested on this medium, 106 (97.2%) produced abundant chlamydospores and three produced few chlamydospores . In contrast, of the 120 C . albicans isolates tested, 111 (92.5%) failed to produce any chlamydospores, whereas the remaining nine isolates produced few chlamydospores . These findings indicate that abundant chlamydospore production on casein agar is a useful test for discriminating between C . dubliniensis and C . albicans.

J Clin Microbiol, 2003 Mar, 41(3), 954 - 9
Use of Fourier-transform infrared spectroscopy for typing of Candida albicans strains isolated in intensive care units; Sandt C et al.; Comparative studies of Candida albicans strains are essential for proving cross-infections in epidemiological investigations . Typing of C . albicans strains is mainly based on genotypic methods . Fourier-transform infrared (FTIR) spectroscopy is described in this study as a novel phenotypic approach to the typing of C . albicans . The first step in the approach was the standardization of sample preparation (culture conditions and sampling parameters) and acquisition and classification parameters (spectral acquisition, spectral window selection, classification algorithm, and heterogeneity threshold) . The second step consisted of validating the established parameters with a set of 79 strains of C . albicans isolated over 4 months from nine patients hospitalized in two intensive care units . Strains were isolated from multiple anatomical sites with repeated sampling . FTIR spectroscopy results were compared to randomly amplified polymorphic DNA (RAPD) results; this analysis showed that the amplification patterns of strains isolated from a given patient were identical and that different patients had different profiles . FTIR spectroscopy data were analyzed by hierarchical clustering performed with the second-derivative spectra . This classification revealed nine groups, one per patient . Only one spectrum out of 79 was misclassified by the FTIR spectroscopy method . RAPD and FTIR spectroscopy results were in good agreement, showing that, when nosocomial candidiasis transmission is suspected and urgent information is needed, this technique may be useful as a quick identification tool to give solid clues before confirmation by a genotypic method.

Microb Pathog, 2003 Feb, 34(2), 103 - 13
Role of complement C5 and T lymphocytes in pathogenesis of disseminated and mucosal candidiasis in susceptible DBA/2 mice; Ashman RB et al.; The aims of the study were to compare the pathogenesis of Candida albicans infection in various organs and anatomical regions of C5-deficient (DBA/2) and C5-sufficient (BALB/c) mice, and to evaluate the importance of complement C5 and T lymphocytes as factors that determine host susceptibility or resistance . The kidneys of DBA/2 mice showed higher colonisation and more severe tissue damage than those of BALB/c, but infection at other sites, including oral and vaginal mucosa, was generally similar in the two strains . Passive transfer of C5-sufficient serum into DBA/2 mice decreased the fungal burden in the kidney, and prolonged survival of the reconstituted animals . Depletion of CD4(+) and/or CD8(+) cells did not exacerbate either systemic or mucosal infection when compared to controls, and passive transfer of splenocytes from infected donors caused only a small and transient reduction in numbers of yeasts recovered from the kidney of sub-lethally infected recipients . It is concluded that the acute susceptibility of the kidneys in this mouse strain is due to C5 deficiency expressed on a susceptible genetic background . T lymphocytes, however, appear to have minimal influence on recovery from systemic infection with this isolate of C . albicans.

Mol Microbiol, 2003 Mar, 47(6), 1637 - 51
Candida albicans binds human plasminogen: identification of eight plasminogen-binding proteins; Crowe JD et al.; Several microbial pathogens augment their invasive potential by binding and activating human plasminogen to generate the proteolytic enzyme plasmin . Yeast cells and cell wall proteins (CWP) of the human pathogenic fungus Candida albicans bound plasminogen with a K(d) of 70 +/- 11 nM and 112 +/- 20 nM respectively . Bound plasminogen could be activated to plasmin by mammalian plasminogen activators; no C . albicans plasminogen activator was detected . Binding of plasminogen to CWP and whole cells was inhibited by epsilon ACA, indicating that binding was predominantly to lysine residues . Candida albicans mutant strains defective in protein glycosylation did not show altered plasminogen binding, suggesting that binding was not mediated via a surface lectin . Binding was sensitive to digestion by basic carboxypeptidase, implicating C-terminal lysine residues in binding . Proteomic analysis identified eight major plasminogen-binding proteins in isolated CWP . Five of these (phosphoglycerate mutase, alcohol dehydrogenase, thioredoxin peroxidase, catalase, transcription elongation factor) had C-terminal lysine residues and three (glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase and fructose bisphosphate aldolase) did not . Activation of plasminogen could potentially increase the capacity of this pathogenic fungus for tissue invasion and necrosis . Although surface-bound plasmin(ogen) degraded fibrin, no direct evidence for a role in invasion of endothelial matrix or in penetration and damage of endothelial cells was found.

Mol Microbiol, 2003 Mar, 47(6), 1601 - 12
The glyoxylate cycle is required for temporal regulation of virulence by the plant pathogenic fungus Magnaporthe grisea; Wang ZY et al.; We describe the isolation and characterization of ICL1 from the rice blast fungus Magnaporthe grisea, a gene that encodes isocitrate lyase, one of the principal enzymes of the glyoxylate cycle . ICL1 shows elevated expression during development of infection structures and cuticle penetration, and a targeted gene replacement showed that the gene is required for full virulence by M . grisea . In particular, we found that the prepenetration stage of development, before entry into plant tissue, is affected by loss of the glyoxylate cycle . There is a delay in germination, infection-related development and cuticle penetration in Delta icl1 mutants . Recent reports have shown the importance of the glyoxylate cycle in the virulence of the human pathogenic fungus Candida albicans and the bacterial pathogen Mycobacterium tuberculosis . Our results indicate that the glyoxylate cycle is also important in this plant pathogenic fungus, demonstrating the widespread utility of the pathway in microbial pathogenesis.

Mol Microbiol, 2003 Mar, 47(6), 1523 - 43
Stage-specific gene expression of Candida albicans in human blood; Fradin C et al.; The pathogenic fungus Candida albicans commonly causes mucosal surface infections . In immunocompromised patients, C . albicans may penetrate into deeper tissue, enter the bloodstream and disseminate within the host causing life-threatening systemic infections . In order to elucidate how C . albicans responds to the challenge of a blood environment, we analysed the transcription profile of C . albicans cells exposed to human blood using genomic arrays and a cDNA subtraction protocol . By combining data obtained with these two methods, we were able to identify unique sets of different fungal genes specifically expressed at different stages of this model that mimics bloodstream infections . By removing host cells and incubation in plasma, we were also able to identify several genes in which the expression level was significantly influenced by the presence of these cells . Differentially expressed genes included those that are involved in the general stress response, antioxidative response, glyoxylate cycle as well as putative virulence attributes . These data point to possible mechanisms by which C . albicans ensures survival in the hostile environment of the blood and how the fungus may escape the bloodstream as an essential step in its systemic dissemination.

J Reprod Med, 2003 Feb, 48(2), 63 - 8
Clinical resistance of recurrent Candida albicans vulvovaginitis to fluconazole in the presence and absence of in vitro resistance; MacNeill C et al.; OBJECTIVE: To determine if failure of recurrent Candida albicans vulvovaginitis to respond clinically to fluconazole is related to in vitro mycologic resistance . STUDY DESIGN: We compared clinical response to fluconazole with culture and sensitivity data in all cases of recurrent C albicans vulvovaginitis referred to our clinic over an 18-month period . RESULTS: Of 52 patients referred to us with recurring vulvovaginitis, 10 were C albicans culture positive . All 10 had previously responded to fluconazole but subsequently failed fluconazole therapy . All were euglycemic and HIV negative . In 3 of the 10 isolates, the mean inhibitory concentration for fluconazole was > 64 micrograms/mL . The history of response to fluconazole in the 7 patients with susceptible isolates was indistinguishable from that of the 3 with resistant isolates . Five of the 10 patients were given multiagent antifungal therapy . Of 4 patients available for long-term follow-up in this group, all had negative fungal cultures . In contrast, 4 evaluable patients who received maintenance azole therapy were C albicans culture positive at long-term follow-up . CONCLUSION: Recurrent C albicans vulvovaginitis can display clinical resistance to fluconazole that correlates with in vitro resistance in only some cases . We postulate that aberrant host response may play a role in the failure to control fungal colonization with a single fungistatic agent.

J Med Microbiol, 2003 Mar, 52(Pt 3), 247 - 9
Effect of antimycotic agents on the activity of aspartyl proteinases secreted by Candida albicans; Schaller M et al.; The inhibitory effect of human immunodeficiency virus (HIV) proteinase inhibitors amprenavir and saquinavir and antifungal agents terbinafine, ketoconazole, amphotericin B and ciclopiroxolamine on aspartyl proteinases (Saps) secreted by Candida albicans was tested in an in vitro spectophotometric assay . As expected, both HIV proteinase inhibitors showed a significant inhibitory effect on Sap activity, which was comparable to that of the classical aspartyl proteinase inhibitor pepstatin A (P < 0.001) . Antifungal drugs such as ketoconazole, terbinafine and amphotericin B had no, or only minor, inhibitory effects on proteolytic activity . In contrast, a significant reduction in Sap activity could be demonstrated during treatment with the antifungal agent ciclopiroxolamine (P < 0.001) . These results point to a multiple effect of this antimycotic agent and might explain the reduced adherence of C . albicans to human epithelial cells at subinhibitory doses.

Phytochemistry, 2003 Mar, 62(5), 747 - 51
Antiplasmodial and antifungal activities of iridal, a plant triterpenoid; Benoit-Vical F et al.; Iridal, a triterpenoidic compound extracted from Iris germanica L., was previously shown to have an interesting activity on two cultured human tumor cell lines (A2780 and K562) . In the present work, this same product was tested in vitro on Plasmodium falciparum chloroquine-resistant and -sensitive strains, in vivo on P . vinckei, and on some Candida albicans and C . parapsilosis strains too . The IC(50) obtained in vitro on human malaria strain ranged from 1.8 to 26.0 microg/ml and the ED(50) in vivo is about 85 mg/kg/day by intraperitoneal route . The minimal inhibitory concentrations were higher than to 50 microg/ml, whatever the strain of yeast tested . This product presents an antiplasmodial activity similar to that obtained with extracts from the plant Azadirachta indica classically taken as reference in malaria phytomedicine . Conversely iridal shows no important antifungal activity . The specific activity of iridal on human malaria parasite and on tumor cell lines is discussed.

Yonsei Med J, 2003 Feb, 44(1), 110 - 8
Search for genes potentially related to germ tube formation in Candida albicans by differential-display reverse transcription polymerase chain reaction; Park JY et al.; Candida albicans exhibits the ability to grow in either a yeast or a mycelia form in response to different environmental factors . The mycelia form, found in infected tissues, is important as a virulence factor in the adherence of the organism to the host epithelium . In vitro, the morphological transition can be induced by environmental shifts in the growing conditions, or by a variety of exogenous factors, including ambient pH, nutritional status and temperature . The differential-display reverse transcription polymerase chain reaction (DDRT-PCR) is a powerful technique for comparing gene expression between cell types, stages of development or differentiation . Hyphae related genes were identified and characterized using a PCR-based differential display . Candida albicans formed a germ tube when cultured in rabbit serum, RPMI 1640 medium or 39 degrees C-YPD medium . We gained 21 cDNA bands showing a different expression pattern from that of the uninduced culture . DNA was extracted from the same location of the isolated bands, and PCR was performed under the same conditions, which reamplified the PCR product, showing the specific expression patterns according to the culture conditions . We cloned 18 germ tube-related cDNA clones (inserts average size is 80 - 700 bp) and sequenced them . The nucleotide sequences of the 18 clones were identified through in the present study from GenBank, and were found to have the accession number (AF405213-AF405230) . We could not find any nucleotide sequence having a high homology with these clones . This study could form a part of the projects in the search for genes related to the germ tube formation of C . albicans.

Forsch Komplementarmed Klass Naturheilkd, 2002 Dec, 9(6), 346 - 51
Comparative investigation of the antimicrobial activity of PADMA 28 and selected European herbal drugs; Weseler A et al.; OBJECTIVE: PADMA 28 is a multicompound preparation of 20 herbs, calcium sulphate, and camphor, derived from Tibetan medicine . It is usually used in the treatment of peripheral circulatory disorders, accompanied by the symptoms tingling, formication, heaviness and tenseness in arms and legs, numbness in hands and feet, and cramps in the calf . Recently, the question of whether appropriate preparations of PADMA 28 also exhibit antibacterial and antimycotic activity has often been raised . As there are as yet no experimental findings that answer this question, an in vitro study was carried out . In a parallel survey we investigated the antimicrobial properties of 5 herbal drugs which are commonly used in the traditional European folk medicine for the topical treatment of mild skin infections, wounds and eczematous skin lesions . METHODS: The minimum inhibitory concentrations (MIC) and the minimum bactericidal concentrations (MBC) of alcohol-based (tinctures) and aqueous (teas) herbal drug preparations were determined in vitro by a broth microdilution method for 5 Gram-positive and 5 Gram-negative bacteria, as well as the yeast Candida albicans . RESULTS: The aqueous and alcohol-based PADMA 28 preparations as well as the corresponding preparations of the European herbal drugs showed an antibacterial effect against Gram-positive bacteria in vitro . These bacteria revealed a somewhat higher sensitivity to the teas prepared from the European herbal drugs (MIC: 1.3-20.0 mg/ml) than to the aqueous preparations of PADMA 28 (MIC: 5.0-40.0 mg/ml) . The better antibacterial activity of the European herbal drugs is probably based on their relatively high amount of tanning agents . On the other hand, all tested plant preparations inhibited not at all or only insufficiently the growth of the Gram-negative bacteria tested and that of Candida albicans . The ethanolic PADMA 28 tinctures showed an improved inhibitory effect on the Gram-positive bacteria (MIC: 0.38-1.51% tincture or 0.38-1.51 mg PADMA 28/ml) compared with the aqueous preparations; this effect is comparable to the ethanolic tinctures of the tested European herbal drugs (MIC: 0.4-1.6/3.2% tincture or 0.4-1.6/3.2 mg herbal drug/ml) . CONCLUSION: All tested tea preparations and alcoholic tinctures of PADMA 28 as well as those of the selected European herbal drugs exhibited evident antibacterial effects against Gram-positive bacteria in vitro . On the other hand, except for Klebsiella pneumoniae, all Gram-negative bacteria tested and the yeast Candida albicans were insensitive against the different aqueous and alcohol-based plant extracts.

Dermatol Surg, 2003 Mar, 29(3), 255 - 60
A prospective trial of fungal colonization after laser resurfacing of the face: correlation between culture positivity and symptoms of pruritus; Alam M et al.; BACKGROUND: After full-face laser resurfacing of the face, patients often complain of pruritus, which may be intense . It has been suggested that some cases of postresurfacing pruritus may be associated with subclinical fungal infection . OBJECTIVE: To determine whether intense pruritus after laser resurfacing of the face is correlated with simultaneous fungal growth of the treated skin . METHODS: Twelve adult female patients undergoing combined full-face laser resurfacing with CO2 and erbium:YAG lasers for chronic photodamage or acne scarring were enrolled in a prospective study . Fungal cultures were obtained by swabbing the facial skin of each patient immediately before, 3 days after, and 6 days after the laser procedure . At the same points in time, investigators completed objective assessments of the patients' facial skin, and patients reported the sensations that they were experiencing . RESULTS: Six patients (50%) complained of significant pruritus (3 or greater on a scale of 0 to 5) . In four of the six cases (67%), at least one of the three fungal cultures obtained grew fungal organisms, including Candida albicans (2 cultures), Candida parapsilosis, Aureobasidium pullulans, and Fusarium species . In no instances did culture positivity occur in the absence of significant pruritus . A statistically significant relationship (P=0.0143) was found to exist between at least one of the three cultures being positive and the emergence of significant posttreatment pruritus . Physician ratings of clinical signs did not correlate with patient reports of pruritus, other symptoms, or culture positivity . CONCLUSION: Colonization or subclinical infection with fungi, particularly Candida spp., may be associated with significant postlaser resurfacing pruritus . Antifungal prophylaxis may mitigate this discomfort . Further research is required to confirm and expand these results.

ASDC J Dent Child, 2002 Sep-Dec, 69(3), 289 - 92, 235
Prevalence of Candida albicans in oral cavities and root canals of children; Akdeniz BG et al.; The aim of this study was to examine the prevalence of C . albicans in the oral cavities and root canals of children . Twenty healthy and caries-free children and 13 children with caries, were screened . Imprint samples and sterile paper points were used to obtain the samples from oral cavities and root canals respectively . The production of germ tubes and the development of chlamydospores identified yeast cultures . Sixty-nine percent of children with caries and 5% of caries-free children were found to be Candida carriers . The difference in candidal prevalence between two groups was significant (p < 0.05) . Sixty-one point five percent of children were positive for Candida in the root canal . Since, increase in the C . albicans in the oral cavity provides a potential source of the fungus particularly when resistance falls below a certain threshold, attention to strategies for the reduction of this pervasive and persistent pathogen becomes important . Therefore, reduction of caries and or introduction of antifungal agents during root canal treatment of children may be offered.

Curr Genet, 2003 Mar, 42(6), 339 - 43 Epub 2003 Jan 30.
An improved transformation protocol for the human fungal pathogen Candida albicans; Walther A et al.; Commonly used protocols for the transformation of the dimorphic human fungal pathogen Candida albicans rely on established methods for the yeast Saccharomyces cerevisiae . With respect to transformation efficiency, however, there is a great difference between these two organisms when using the lithium acetate procedure . Here we present a modified version of this protocol for use with C . albicans . Among the different parameters tested, two turned out to be particularly relevant and, when combined, resulted in an up to 10-fold increase in transformation efficiency (400-500 integrative transformants) compared with previous protocols: first, adjusting the heat shock applied to the cells to 44 degrees C for C . albicans instead of 42 degrees C for S . cerevisiae and, second, treating C . albicans cells with lithium acetate in an overnight incubation instead of for 30 min as used for S . cerevisiae . With these modifications, the lithium acetate procedure becomes a very efficient and reliable tool for C . albicans transformation.

J Microbiol Methods, 2003 Apr, 53(1), 11 - 5
Enzymatic differentiation of Candida parapsilosis from other Candida spp . in a membrane filtration test; Bauters TG et al.; A previously reported enzyme assay on a membrane filter using 4-methylumbelliferyl (4-MU)-N-acetyl-beta-D-galactosaminide, -phosphate and -pyrophosphate as substrates for the differentiation of four Candida spp . has been extended to Candida parapsilosis . The substrate 4-MU-beta-D-glucoside was hydrolyzed by 28 test strains of this species but to a variable extent by seven other yeasts also . For a full enzymatic differentiation of C . parapsilosis from other medical yeasts, a battery of six reactions was required . Of 71 C . parapsilosis positive clinical samples, 4.2% gave a false negative result due to overgrowth by Candida albicans . The present assay is more rapid than a described spectrofluorometric determination of beta-D-glucosidase in a broth, i.e., 9-11 h versus up to >48 h.

J Nat Prod, 2003 Feb, 66(2), 272 - 5
Eudistomins W and X, two new beta-carbolines from the micronesian tunicate Eudistoma sp; Schupp P et al.; Chemical investigation of the Micronesian ascidian Eudistoma sp . afforded two new eudistomin congeners, which were designated eudistomins W (1) and X (2) . The structures of the new compounds were unambiguously established on the basis of NMR spectroscopic ((1)H, (13)C, COSY, (1)H detected direct, and long-range (13)C-(1)H correlations) and mass spectrometric (EI and ESIMS) data . Compound 2 exhibited antibiotic activity toward Bacillus subtilis, Staphyloccocus aureus, and Escherichia coli and was also found to be fungicidal against Candida albicans in an agar diffusion assay . Compound 1 was selectively active against C . albicans but showed no antibacterial activity.

Med Hypotheses, 2003 Feb, 60(2), 188 - 9
Candida albicans and selenium; Reid GM; Although low selenium levels have been recorded in infants, no specific human disorder has been linked to low selenium status . The incidence of thrush, the common enteric fungal infection caused by Candida albicans, has increased markedly with antibiotic therapy and research has provided evidence that its colonization leads to competition for Coenzyme Q10 (CoQ10) in the host . Furthermore it is now known that ubiquinones are essential in heart muscle for oxidative phosphorylation in the mitochondrial respiratory chain and considered that glutathione peroxidase (GSHPx) in the mitochondria protects ubiquinone from oxidation.

J Biol Chem, 2003 May 9, 278(19), 17228 - 35 Epub 2003 Feb 24.
Aha1 binds to the middle domain of Hsp90, contributes to client protein activation, and stimulates the ATPase activity of the molecular chaperone; Lotz GP et al.; The ATP-dependent molecular chaperone Hsp90 is an essential and abundant stress protein in the eukaryotic cytosol that cooperates with a cohort of cofactors/cochaperones to fulfill its cellular tasks . We have identified Aha1 (activator of Hsp90 ATPase) and its relative Hch1 (high copy Hsp90 suppressor) as binding partners of Hsp90 in Saccharomyces cerevisiae . By using genetic and biochemical approaches, the middle domain of Hsp90 (amino acids 272-617) was found to mediate the interaction with Aha1 and Hch1 . Data base searches revealed that homologues of Aha1 are conserved from yeast to man, whereas Hch1 was found to be restricted to lower eukaryotes like S . cerevisiae and Candida albicans . In experiments with purified proteins, Aha1 but not Hch1 stimulated the intrinsic ATPase activity of Hsp90 5-fold . To establish their cellular role further, we deleted the genes encoding Aha1 and Hch1 in S . cerevisiae . In vivo experiments demonstrated that Aha1 and Hch1 contributed to efficient activation of the heterologous Hsp90 client protein v-Src . Moreover, Aha1 and Hch1 became crucial for cell viability under non-optimal growth conditions when Hsp90 levels are limiting . Thus, our results identify a novel type of cofactor involved in the regulation of the molecular chaperone Hsp90.

Antimicrob Agents Chemother, 2003 Mar, 47(3), 1068 - 71
In vitro activities of caspofungin compared with those of fluconazole and itraconazole against 3,959 clinical isolates of Candida spp., including 157 fluconazole-resistant isolates; Pfaller MA et al.; Caspofungin is an echinocandin antifungal agent with broad-spectrum activity against Candida and Aspergillus spp . The in vitro activities of caspofungin against 3,959 isolates of Candida spp . obtained from over 95 different medical centers worldwide were compared with those of fluconazole and itraconazole . The MICs of the antifungal drugs were determined by broth microdilution tests performed according to the NCCLS method using RPMI 1640 as the test medium . Caspofungin was very active against Candida spp . (MIC at which 90% of the isolates were inhibited {MIC(90)}, 1 micro g/ml; 96% of MICs were < or =2 micro g/ml) . Candida albicans, C . dubliniensis, C . tropicalis, and C . glabrata were the most susceptible species of Candida (MIC(90), 0.25 to 0.5 micro g/ml), and C . guilliermondii was the least susceptible (MIC(90), >8 micro g/ml) . Caspofungin was very active against Candida spp., exhibiting high-level resistance to fluconazole and itraconazole (99% of MICs were < or =1 micro g/ml) . These results provide further evidence for the spectrum and potency of caspofungin activity against a large and geographically diverse collection of clinically important isolates of Candida spp.

Antimicrob Agents Chemother, 2003 Mar, 47(3), 956 - 64
Ergosterol biosynthesis inhibitors become fungicidal when combined with calcineurin inhibitors against Candida albicans, Candida glabrata, and Candida krusei; Onyewu C et al.; Azoles target the ergosterol biosynthetic enzyme lanosterol 14alpha-demethylase and are a widely applied class of antifungal agents because of their broad therapeutic window, wide spectrum of activity, and low toxicity . Unfortunately, azoles are generally fungistatic and resistance to fluconazole is emerging in several fungal pathogens . We recently established that the protein phosphatase calcineurin allows survival of Candida albicans during the membrane stress exerted by azoles . The calcineurin inhibitors cyclosporine A (CsA) and tacrolimus (FK506) are dramatically synergistic with azoles, resulting in potent fungicidal activity, and mutant strains lacking calcineurin are markedly hypersensitive to azoles . Here we establish that drugs targeting other enzymes in the ergosterol biosynthetic pathway (terbinafine and fenpropimorph) also exhibit dramatic synergistic antifungal activity against wild-type C . albicans when used in conjunction with CsA and FK506 . Similarly, C . albicans mutant strains lacking calcineurin B are markedly hypersensitive to terbinafine and fenpropimorph . The FK506 binding protein FKBP12 is required for FK506 synergism with ergosterol biosynthesis inhibitors, and a calcineurin mutation that confers FK506 resistance abolishes drug synergism . Additionally, we provide evidence of drug synergy between the nonimmunosuppressive FK506 analog L-685,818 and fenpropimorph or terbinafine against wild-type C . albicans . These drug combinations also exert synergistic effects against two other Candida species, C . glabrata and C . krusei, which are known for intrinsic or rapidly acquired resistance to azoles . These studies demonstrate that the activity of non-azole antifungal agents that target ergosterol biosynthesis can be enhanced by inhibition of the calcineurin signaling pathway, extending their spectrum of action and providing an alternative approach by which to overcome antifungal drug resistance.

Ann Allergy Asthma Immunol, 2003 Feb, 90(2), 259 - 64
The syndrome of chronic mucocutaneous candidiasis with selective antibody deficiency; Kalfa VC et al.; BACKGROUND: Most patients with chronic mucocutaneous candidiasis (CMC) have a selective defect of cell-mediated immunity against Candida albicans (as demonstrated by cutaneous anergy and decreased lymphoproliferative responses to Candida antigen) and intact antibody responses . Many CMC patients also develop infections with other organisms, suggesting a more extensive immunologic defect . OBJECTIVES: The aim of this study was to describe a patient with CMC and selective antibody deficiency and identify eight similar previously reported patients . DATA SOURCES: Relevant articles in the English language derived from searching the MEDLINE database were used . RESULTS: We describe an 18-year-old male patient who was identified with CMC as an infant and later developed immunoglobulin (Ig)G2, IgG4, and IgA deficiency at age 12 associated with poor antibody responses to vaccine antigens . We have identified eight other previously reported CMC patients with selective antibody deficiencies and bacterial infections . IgG2 deficiency was present in all nine patients, and was associated with IgG4 deficiency in 8 patients and IgA deficiency in 3 patients . Six patients had poor or absent antibody responses to pneumococcal polysaccharide vaccine, and all nine patients developed severe recurrent lung infections . CONCLUSIONS: We suggest that these cases represent a distinct phenotype of CMC and should be studied for common histocompatibility leukocyte antigen types and molecular defects.

Phytother Res, 2003 Feb, 17(2), 190 - 3
Antifungal, antioxidant and larvicidal activities of compounds isolated from the heartwood of Mansonia gagei; Tiew P et al.; Eleven compounds isolated from the heartwood of Mansonia gagei were tested for their antifungal activities against Cladosporium cucumerinum and Candida albicans, as well as for their larvicidal activities against Aedes aegypti and radical scavenging properties in a DPPH assay . Mansonone C (4) was found to be the most interesting compound with antifungal activities against Cladosporium cucumerinum and Candida albicans as well as for its larvicidal properties against Aedes aegypti . Mansonone E (5) was active against Cladosporium cucumerinum and Candida albicans . Two coumarin derivatives, mansorin A (1) and mansorin B (2) were also found to be active against Cladosporium cucumerinum, while mansonone N (9) was the only isolated product to show radical scavenging properties .

Phytother Res, 2003 Feb, 17(2), 183 - 6
The in vivo antifungal activity of the aqueous extract from Nigella sativa seeds; Khan MA et al.; The effect of an aqueous extract of Nigella sativa seeds was studied on candidiasis in mice . An intravenous inoculum of Candida albicans produced colonies of the organism in the liver, spleen and kidneys . Treatment of mice with the plant extract (6.6 mL/kg equivalent to 5 mg of estimated protein, once daily for 3 days) 24 h after the inoculation caused a considerable inhibitory effect on the growth of the organism in all organs studied . A 5-fold decrease in Candida in kidneys, 8-fold in liver and 11-fold in spleen was observed in the groups of animals post-treated with the plant extract . Histopathological examination of the respective organs confirmed these findings . These results indicate that the aqueous extract of Nigella sativa seeds exhibits inhibitory effect against candidiasis and this study validates the traditional use of the plant in fungal infections .

Phytother Res, 2003 Feb, 17(2), 155 - 9
Cytotoxic activity of chalcones isolated from lonchocarpus sericeus (pocr.) kunth; Cunha GM et al.; In the present study, it was demonstrated that the hexanic extract obtained from the roots of Lonchocarpus sericeus and one of its major components, derricin, but not lonchocarpin, show cytotoxic activity to fertilized sea urchin eggs . Both inhibited the first cleavage of sea urchin eggs in a dose-dependent manner with an IC(50) of 30.4 (26.2-35.3) microgram/mL for the crude extract and 51.2 (42.1-62.3) microgram/mL for derricin (n = 6, in both cases) . Furthermore, the hexanic extract of L . sericeus, and the isolated compounds, derricin and lonchocarpin showed cytotoxicity against CEM leukaemic cell line (IC(50) = 17.6 (13.7-22.6), 13.0 (12.0-14.0) and 10.4 (5.6-19.1) microgram/ml (n = 6), respectively) . When these substances (6.25 to 125 microgram/25 microL) were examined for antimicrobial activity against Escherichia coli, Staphylococcus aureus and Candida albicans, none of them were found to be active . Neither the hexanic extract, nor the isolated compounds derricin and lonchocarpin (0.5 to 250 microgram/mL) presented hemolytic activity . These results indicated a possible antimitotic activity of L . sericeus crude extract and their major constituents .

J Clin Virol, 2003 Feb, 26(2), 247 - 63
Defective phagocytosis by human monocyte/macrophages following HIV-1 infection: underlying mechanisms and modulation by adjunctive cytokine therapy; Kedzierska K et al.; Defective immunological function of cells of the macrophage lineage contributes considerably to the pathogenesis of HIV-1 infection . Impairment of phagocytosis of opportunistic pathogens such as Mycobacterium avium complex (MAC), Pneumocystis carinii, Toxoplasma gondii or Candida albicans by peripheral blood monocytes, tissue macrophages and monocyte-derived macrophages following in vivo and in vitro HIV-1 infection is well documented . The development of opportunistic infections due to these pathogens in HIV-infected individuals at late stages of disease is attributed to defective monocyte/macrophage function . The mechanisms whereby HIV-1 impairs phagocytosis are not well known . A number of phagocytic receptors normally mediate engulfment of specific opportunistic pathogens by cells of macrophage lineage; distinct mechanisms are triggered by pathogen-receptor binding to promote cytoskeletal rearrangements and engulfment . This review focuses on the signalling events occurring during Fcgamma receptor- and complement receptor-mediated phagocytosis, and considers the mechanisms by which HIV-1 inhibits those signalling events . Since macrophage function is enhanced by cytokines such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (IFN-gamma), the use of these immunomodulators is of potential interest as adjunctive immunotherapy in immunosuppressed individuals . In this review we present examples of clinical applications of GM-CSF and IFN-gamma therapy for the treatment of opportunistic infections in HIV-infected individuals receiving antiretroviral drugs .

J Infect Dis, 2003 Feb 15, 187(4), 710 - 3 Epub 2003 Jan 29.
Suppression by Candida albicans beta-glucan of cytokine release from activated human monocytes and from T cells in the presence of monocytes; Nakagawa Y et al.; The effect of a soluble beta-glucan from Candida albicans (CSBG) on cytokine production by cultured human peripheral blood mononuclear cells (PBMC) was assessed . CSBG induced a slight increase in the spontaneous release of proinflammatory cytokines, such as interleukin (IL)-6, but significantly suppressed endotoxin-induced IL-6 production in cultures of PBMC and monocytes isolated from PBMC . CSBG also suppressed the release of type 1 cytokines, IL-2, and interferon-gamma . These findings suggest that CSBG suppresses monocyte functions directly and thus suppresses T cell function indirectly . CSBG may play a role in the development of candidiasis.

Trends Microbiol, 2003 Feb, 11(2), 69 - 73
URA3 as a selectable marker for disruption and virulence assessment of Candida albicans genes; Staab JF et al.; The ability to generate isogenic sets of strains with mutations in a gene of interest but not in other genes by repeated use of the URA3 marker (Ura-blaster methodology) has advanced our understanding of the relationships between gene structure and function in Candida albicans . Common applications of Ura-blaster technology result in different genomic positions for the URA3 gene in strains complemented for the gene of interest compared with mutant strains . Studies using animal models of systemic candidiasis pointed to possible differences in URA3 gene expression, depending on its genomic location, which confounded interpretation of the role of the gene of interest in lethality . Positional effects on URA3 expression can be avoided by placement at a common locus in all strains used for comparison.

Indian J Exp Biol, 2002 Jul, 40(7), 785 - 90
Effect of phosphocreatine on H+ extrusion, pHi and dimorphism in Candida albicans; Manzoor N et al.; Candida albicans is an opportunistic pathogen . Its proliferation in human hosts is believed to be controlled by immunologic mechanisms . The plasma membrane of the fungus possesses an H(+)-ATPase (PM-ATPase) which actively extrudes protons to generate an electrochemical gradient which is used in co-transport of nutrients . This ATPase is associated with the growth, dimorphism and pathogenicity of the fungus . The physiological concentration of phosphocreatine (PCr) is 20-35 mM in skeletal muscles . H(+)-extrusion in Candida cells was strongly inhibited by PCr; 44% at 20 mM and 69% at 40 mM . H(+)-extrusion was stimulated 6.2-fold in the presence of 10 mM glucose . This glucose stimulated extrusion was inhibited significantly by PCr; 36% at 20 mM and 53% at 40 mM . The intracellular pH pattern of cells destined to differentiate was greatly altered in the presence of PCr . Evagination time for control cells was between 90-120 min . PCr, delayed dimorphism, reduced the population of cells differentiating to hyphae and also reduced the length of hyphae after each time interval . Only 60% differentiation was observed with 10 mM PCr and 40% for higher PCr concentration even after 210 min . Direct interaction of PM-ATPase and PCr has been demonstrated by difference spectrum measurement employing stopped flow spectrophotometer . It can be concluded that PCr may be playing a significant role in checking growth and pathogenesis of C . albicans.

Microbiol Immunol, 2002, 46(12), 881 - 3
Extracellular proteinase and phospholipase activity of three genotypic strains of a human pathogenic yeast, Candida albicans; Sugita T et al.; Strains of a human pathogenic yeast, Candida albicans, have (A) intronless, (B) intron-containing, and (C) a mixture of intron-containing and intronless 26S rRNA genes . To elucidate the significance of these three genotypes in pathogenesis, we measured two major virulence factors, extracellular proteinase and phospholipase activity, in 56 clinical isolates of C . albicans, and investigated the relationship between genotype and enzymatic activity . The genotype B strains had significantly higher proteinase and phospholipase activities than genotypes A or C . These results suggest that to understand the pathogenesis of C . albicans, the genotypes should be considered.

Eur J Med Chem, 2003 Jan, 38(1), 75 - 87
Synthesis and antifungal activity of new 1-halogenobenzyl-3-imidazolylmethylindole derivatives; Na YM et al.; A series of 1-benzyl-3-(imidazol-1-ylmethyl)indole derivatives 35-46 were prepared under mild reaction conditions and tested for their antifungal activity . Pharmacomodulation at N(1), C(2) and C(5) of the indole ring and at the level of the alkyl chain (R(1)) was carried out starting from the corresponding 3-acylindoles 6, 7 or 3-formylindoles 11-22 . Target imidazolyl compounds 35-46 were obtained in satisfactory yields by CO(2) elimination from the intermediate carbamates . All of the compounds were evaluated in vitro against two human fungal pathogens, Candida albicans (CA980001) and Aspergillus fumigatus (AF980003); amphotericin B, fluconazole and itraconazole were used as references . Seven out of 27 compounds (35b, 35e, 35g, 35h, 36a, 38a and especially 40a) exerted significant antifungal activity against C . albicans, with MIC in the range of 1-6 microg mL(-1) . As regards inhibitory activity against A . fumigatus, the MIC figures of most of our compounds were in excess of 20 microg mL(-1) in contrast to the reference drugs, amphotericin B and itraconazole, whose MIC(90) and MIC(80) values were 0.14 and 0.50 microg mL(-1), respectively . The most potent compound, 45a, exhibited MIC value (8 +/- 1 microg mL(-1)) 16-fold higher than that of itraconazole .

Eur J Med Chem, 2003 Jan, 38(1), 27 - 36
Design and synthesis of some substituted 1H-pyrazolyl-thiazolo{4,5-d}pyrimidines as anti-inflammatory-antimicrobial Agents; Bekhit AA et al.; The synthesis of two novel series of structurally related 1H-pyrazolyl derivatives of thiazolo{4,5-d}pyrimidines is described . All the newly synthesised compounds were examined for their in vivo anti-inflammatory activity in two different bioassays namely; cotton pellet-induced granuloma and carrageenan-induced paw edema in rats . The in vitro inhibitory activity of the most active compounds towards human COX-1 and COX-2 enzymes was also estimated . In addition, the ulcerogenic effects and acute toxicity (LD(50)) values of these compounds 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 results revealed that compounds 5a, 9a, 9b, 10b and 12a exhibited anti-inflammatory activity comparable to that of indomethacin in both local and systemic in vivo animal models with no or minimal ulcerogenic effects (0-10%) and high safety margin (LD(50) > 500 mg kg(-1)) . In addition, most of them displayed appreciable antibacterial activities when compared with ampicillin, especially against S . aureus . Compounds 9a and 12a are the most distinctive derivatives identified in the present study because of their remarkable in vivo and in vitro anti-inflammatory activity in addition to their pronounced antibacterial activities comparable to ampicillin against Gram positive and -negative bacteria . Therefore, they are considered as successful dual anti-inflammatory-antimicrobial candidates .

J Med Chem, 2003 Feb 27, 46(5), 850 - 5
Novel alpha-melanocyte stimulating hormone peptide analogues with high candidacidal activity; Grieco P et al.; alpha-Melanocyte stimulating hormone (alpha-MSH) is an endogenous linear tridecapeptide with potent antiinflammatory effects . We recently demonstrated that alpha-MSH and its C-terminal sequence Lys-Pro-Val (alpha-MSH (11-13)) have antimicrobial effects against two major and representative pathogens: Staphylococcus aureus and Candida albicans . In an attempt to improve the candidacidal activity of alpha-MSH and to better understand the peptide structure-antifungal activity relations, we designed and synthesized novel peptide analogues . Because previous data suggested that antimicrobial effects of alpha-MSH were receptor-mediated, we chose to focus on the sequence alpha-MSH (6-13), which contains the invariant core sequence His-Phe-Arg-Trp (6-9) that is important for binding to the known melanocortin receptors and also contains the sequence Lys-Pro-Val (11-13) that is known to be important for antimicrobial activity . In this structure-activity study, we discovered several compounds that have greater candidacidal activity than alpha-MSH . The peptide {d-Nal-7,Phe-12}-alpha-MSH (6-13) was the most potent of the analogues tested . The present results are very encouraging because they show the great potential of these peptides as a truly novel class of candidacidal compounds.

J Laryngol Otol, 2003 Jan, 117(1), 39 - 42
Otomycosis in Turkey: predisposing factors, aetiology and therapy; Ozcan KM et al.; Otomycosis usually requires long-term treatment and tends to recur . This study was performed on 87 patients with the clinical diagnosis of otomycosis and 20 controls in order to determine the pathogenic agents, predisposing factors and a cost-effective treatment . The predisposing factors included wearing head clothes (74.7 per cent), presence of dermatomycoses (34.5 per cent) and swimming (27.6 per cent) . The most common pathogenic fungus was Aspergillus niger (44.8 per cent) in the otomycosis group . The only isolate was Candida albicans in the control group (2.5 per cent) . We concluded that administration of four per cent boric acid solution in alcohol and frequent suction cleaning of the ear canal might be a cost-effective treatment for otomycosis since 77 per cent of the patients were treated effectively this way . Eighty per cent of the resistant cases had mixed fungal-bacterial infections, and 50 per cent of them had dermatomycoses . These resistant cases were treated by administration of tioconazole ointment.

FEMS Immunol Med Microbiol, 2003 Jan 21, 35(1), 11 - 5
Interaction of sertraline with Candida species selectively attenuates fungal virulence in vitro; Lass-Florl C et al.; This study investigated whether the interaction between isolates of Candida albicans (n=7), Candida parapsilosis (n=3), Candida krusei (n=2), Candida dubliniensis (n=1) and sertraline, a typical selective serotonin reuptake inhibitor, alters candidal virulence . Sertraline treatment of Candida spp . significantly (P<0.05) affected hyphal elongation, phospholipase activity, production of secreted aspartyl proteinases and fungal viability . In addition, monocyte-derived macrophages (MDMs) treated with sertraline reduced inhibition of blastoconidia germination in comparison to MDMs alone . In conclusion, our findings suggest that the interaction between sertraline and Candida spp . may also diminish the virulence properties of this fungal pathogen in vivo.

Biochem Biophys Res Commun, 2003 Feb 21, 301(4), 1099 - 108
Two membrane proteins located in the Nag regulon of Candida albicans confer multidrug resistance; Sengupta M et al.; Pathogenic fungus Candida albicans can efficiently utilize the aminosugar N-acetylglucosamine (GlcNAc) as energy source . Since the mucosal membrane, the site of infection is rich in amino sugars, this specific adaptation is important for the establishment of infection . The genes encoding for the enzymes of the GlcNAc catabolic pathway, GlcNAc kinase (HXK1), GlcNAc-6-phosphate deacetylase (DAC1), and glucosamine-6-phosphate deaminase (NAG1), are present in a cluster, the Nag regulon, which is associated with virulence . In this study, we have characterized two genes, TMP1 and TMP2, present within the Nag regulon, upstream to DAC1 . They encode two membrane associated sugar transporters of the major facilitator superfamily (MFS) . The null mutant of TMP1 and TMP2 is able to grow in GlcNAc, implying that they are not involved in GlcNAc transport . However, it shows increased susceptibility to a number of unrelated antifungal compounds such as cycloheximide, 4-nitroquinoline-N-oxide, and 1-10 phenanthroline . Northern blot analysis revealed that TMP1 and TMP2 are upregulated in response to these drugs, suggesting that they function as multiple drug efflux pumps.

Mycoses, 2003 Feb, 46(1-2), 38 - 41
Tinea pedis observed in Cagliari, Italy, between 1996 and 2000; Aste N et al.; The aim of this study was to verify the incidence of tinea pedis in patients observed in the Department of Dermatology of the University of Cagliari, Italy, in the period from 1996 to 2000 . We examined 722 patients, 536 with lesions and 186 without lesions of the feet and in 169 of them (23.4%) we diagnosed tinea pedis . All patients suffering from tinea pedis belonged to the group with lesions . In the 536 patients with clinical manifestations which were evident to a certain degree, microscope and cultural examination gave positive results for dermatophytes in 169 cases (31.5%) . The most frequently isolated dermatophyte was Trichophyton mentagrophytes (51.5%), followed by Trichophyton rubrum (45.2%) and Epidermophyton floccosum (3.3%) . In the 186 patients without lesions, direct microscope examination was consistently negative while cultural examination showed the growth of sparse colonies of Candida albicans in two cases (1.1%) We analysed distribution by sex, age, residence, occupation and clinical manifestations . Stressing the high frequency of tinea pedis in this region, the discovery of a dermatophytic infection of the feet with an absence of signs and symptoms is an exceptional event.

Mycoses, 2003 Feb, 46(1-2), 19 - 23
Prevalence of Aspergillus fumigatus and other fungal species in the sputum of adult patients with cystic fibrosis; Bakare N et al.; Aspergillus fumigatus is often found in the respiratory tract secretions of patients with cystic fibrosis (CF), although the role of the fungus for progression of pulmonary disease remains unclear . This study aimed to investigate the frequency of A . fumigatus and other fungi in sputum of adult CF patients using different methods for culture and microscopy . Results from the analysis of 369 samples from 94 patients showed that A . fumigatus could be isolated in 45.7% of patients . Other moulds were rare, but the yeast Candida albicans was another frequent isolate, detected in 75.5% of patients . A comparison of different culture media showed no difference between a selective medium developed to specifically inhibit Pseudomonas aeruginosa and a standard fungal culture medium for growth of A . fumigatus, although both were more efficient for detection of fungi than other bacterial culture media . Fluorescent microscopy with calcofluor white was more sensitive for detection of fungal hyphae in undiluted sputum than standard methylene blue staining . This study shows that A . fumigatus and C . albicans have a high frequency in adult CF patients . Microbiological analysis should routinely include methods for specific identification of fungi to monitor for potential complications arising from fungal disease in these patients.

FEMS Microbiol Lett, 2003 Jan 21, 218(1), 195 - 202
MYO2 is not essential for viability, but is required for polarized growth and dimorphic switches in Candida albicans; Woo M et al.; The human fungal pathogen Candida albicans changes from a budding yeast form to a polarized hyphal form in response to various external conditions . Dimorphic switching of C . albicans has been implicated in the development of pathogenicity . Morphogenic transformation requires polarized cell growth and rearrangement of the cytoskeleton . We previously showed that myosins play key roles in the conversion from the bud to the hyphal form of C . albicans by inhibiting myosin activities with 2,3-butanedione-2-monoxime (BDM), a general myosin ATPase inhibitor . In this study we investigated the function of MYO2 in C . albicans using deletion mutants . The amino acid sequence of CaMYO2 shows 60% identity and 77% homology with MYO2 and 54% identity and 70% homology with MYO4 of budding yeast Saccharomyces cerevisiae, suggesting that CaMYO2 is the only class V myosin in C . albicans . Cells in which both CaMYO2 alleles were deleted were viable, suggesting that MYO2 is nonessential in C . albicans . The proliferation of CaMYO2delta cells, however, was sharply decreased . In addition, CaMYO2delta cells showed defects in assembly and polarized localization of F-actin as well as an inability to induce germ tube formation and hyphal growth . The deletion of CaMYO2 also disrupted the shape and migration of the nucleus . These results strongly suggest that CaMYO2 is essential for polarized growth and hyphal transition in C . albicans.

J Chemother, 2002 Dec, 14(6), 627 - 30
Successful treatment of Candida albicans osteomyelitis of the spine with fluconazole and surgical debridement: case report; El-Zaatari MM et al.; A 65-year old diabetic male presented with progressive bone destruction of thoracic spine (T-11&12) with cord compression . Candida albicans was isolated from aspirated materials pre-and intra-operative . Two weeks of fluconazole was given prior to surgical debridement, and fixation of the lesion . C . albicans isolated pre-and 2-weeks after fluconazole treatment were DNA-typed using AP-PCR . MIC was 2-4 mg/l in all isolates tested . The pre-and post treatment isolates had two DNA patterns, indicating the existence of two different strains . Surgical treatment was necessary for patient recovery.

Eukaryot Cell, 2003 Feb, 2(1), 49 - 61
Cell biology of mating in Candida albicans; Lockhart SR et al.; It was recently demonstrated that strains homozygous for either of the mating type-like loci MTLa and MTLalpha of Candida albicans undergo white-opaque switching and that expression of the opaque-phase phenotype greatly enhances mating between strains . Exploiting the latter property to obtain high-frequency mating, we have characterized the cell biology of the mating process of C . albicans . Employing continuous videomicroscopy, computer-assisted three-dimensional reconstruction of living cells, and fluorescence microscopy, we have monitored the mating-associated processes of conjugation, tube formation, fusion, budding, septum formation, and daughter cell development and the spatial and temporal dynamics of nuclear migration and division . From these observations, a model for the stages in C . albicans mating is formulated . The stages include shmooing, chemotropism of conjugation tubes, fusion of tubes and nuclear association, vacuole expansion and nuclear separation in the conjugation bridge, asynchronous nuclear division in the zygote, bud growth, nuclear migration into the daughter cell, septation, and daughter cell budding . Since there was no cytological indication of karyogamy, genetic experiments were performed to assess marker segregation . Recombination was not observed, suggesting that mating takes place in the absence of karyogamy between naturally occurring, homozygous a and alpha strains . This study provides the first description of the cell biology of the mating process of C . albicans.

Eukaryot Cell, 2003 Feb, 2(1), 9 - 18
Cdc24, the GDP-GTP exchange factor for Cdc42, is required for invasive hyphal growth of Candida albicans; Bassilana M et al.; Candida albicans, the most common human fungal pathogen, is particularly problematic for immunocompromised individuals . The reversible transition of this fungal pathogen to a filamentous form that invades host tissue is important for its virulence . Although different signaling pathways such as a mitogen-activated protein kinase and a protein kinase A cascade are critical for this morphological transition, the function of polarity establishment proteins in this process has not been determined . We examined the role of four different polarity establishment proteins in C . albicans invasive growth and virulence by using strains in which one copy of each gene was deleted and the other copy expressed behind the regulatable promoter MET3 . Strikingly, mutants with ectopic expression of either the Rho G-protein Cdc42 or its exchange factor Cdc24 are unable to form invasive hyphal filaments and germ tubes in response to serum or elevated temperature and yet grow normally as a budding yeast . Furthermore, these mutants are avirulent in a mouse model for systemic infection . This function of the Cdc42 GTPase module is not simply a general feature of polarity establishment proteins . Mutants with ectopic expression of the SH3 domain containing protein Bem1 or the Ras-like G-protein Bud1 can grow in an invasive fashion and are virulent in mice, albeit with reduced efficiency . These results indicate that a specific regulation of Cdc24/Cdc42 activity is required for invasive hyphal growth and suggest that these proteins are required for pathogenicity of C . albicans.

Mol Microbiol, 2003 Feb, 47(4), 1029 - 43
Ssn6, an important factor of morphological conversion and virulence in Candida albicans; Hwang CS et al.; Candida albicans, the major fungal pathogen in humans, undergoes morphological conversion from yeasts to filamentous growth forms depending upon various environmental conditions . Here, we have identified a C . albicans gene, namely SSN6, encoding a putative global transcriptional co-repressor that is highly homologous to the Saccharomyces cerevisiae Ssn6 . The isolated C . albicans SSN6 complemented the pleiotropic phenotypes of S . cerevisiae ssn6 mutation, and its expression levels declined significantly in response to a strong true hyphal inducer, serum . The mutant lacking C . albicans Ssn6 displayed a stubby pseudohyphal growth pattern, derepressed filament-specific genes in response to elevated temperature 37 degrees C and failed to develop true hyphae, extensive filamentation and virulence . Such morphological defects of ssn6/ssn6 mutant were not rescued by overexpression of Tup1, Cph1 or Efg1 . Moreover, epistatic analysis showed that, as far as cell morphology was concerned, Ssn6 was epistatic to Tup1 at the higher temperature but that, at the lower temperature, the ssn6/ssn6 tup1/tup1 double mutant grew in a stubby form of pseudohyphae distinct from the phenotypes of either single mutant . Furthermore, overexpression of SSN6 in C . albicans led to enhanced filamentous growth and attenuated virulence . These findings suggest that Ssn6 may function as an activator as well as a repressor of filamentous growth and be a target for candidacidal drugs, as its excess or deficiency resulted in impaired virulence.

Mol Microbiol, 2003 Feb, 47(4), 943 - 59
A screen in Saccharomyces cerevisiae identified CaMCM1, an essential gene in Candida albicans crucial for morphogenesis; Rottmann M et al.; Morphogenesis in Saccharomyces cerevisiae and the pathogenic yeast Candida albicans is governed in part by the same molecular circuits . In S . cerevisiae, FLO11/MUC1 expression has been shown to be modulated by multiple signalling pathways required for pseudohyphal development . We have established a screen in S . cerevisiae to identify regulators of fungal development in C . albicans based on FLO11::lacZ expression as a reporter . This screen identified both known components of the mitogen-activated protein kinase (MAPK) cascade and the cAMP cascade that are important for hyphal development in C . albicans, as well as genes not yet known to be involved in morphogenesis . The Candida homologue of MCM1 is one of the novel factors identified in this screen as being important for morphogenesis . CaMcm1p levels do not vary significantly in different cell types and respond to an autoregulatory feedback mechanism, arguing that CaMcm1p activity is regulated by post-translational modifications . Both overexpression and repression of this essential gene led to the induction of hyphae . Moreover, we found that the expression of HWP1, a hyphae-specific gene, was induced by repression of CaMCM1 . The changes in morphology and HWP1 expression were not the result of a change in expression levels of NRG1 or TUP1, known repressors of hyphal development . Thus, CaMcm1p is a component of a hitherto unknown regulatory mechanism of hyphal growth.

Allergy, 2003 Jan, 58(1), 72 - 7
HLA-DR-dependent increased mannan-induced lymphoproliferative response in atopic eczema dermatitis syndrome; Savolainen J et al.; BACKGROUND: In yeast-sensitive atopic eczema dermatitis syndrome (AEDS), yeast mannan induces highly elevated specific IgE levels and lymphoproliferative responses . In healthy individuals the involvement of both human leukocyte antigen (HLA)-dependent T-cell activation and non-HLA-dependent activation, e.g . by crosslinking of the cell surface mannose receptors, has been suggested . In the present study the HLA dependence and the role of crosslinking in the lymphoproliferative response to mannan in AEDS has been analyzed . METHODS: Twenty patients with AEDS and 12 controls with no history of allergic diseases were included in the study . Mannan from Candida albicans was prepared according to the Cetavlon method . Following isolation using Ficoll-Hypaque, peripheral blood mononuclear cells (PBMC) were incubated with the mannan preparation in the absence and presence of different concentrations of neutralizing anti-HLA antibodies and alpha-methylmannoside for 6 days and proliferative responses were measured by 3H-thymidine incorporation and scintilloradiography . RESULTS: In AEDS patients with elevated mannan-specific serum IgE, the C . albicans mannan induced lymphoproliferation . Mannan-induced lymphoproliferative responses could be inhibited, dose-dependently, by neutralizing anti-HLA-DR, but not anti-HLA-DQ antibodies in AEDS patients and healthy controls . The addition of alpha-methylmannoside, that blocks binding to mannose receptors, inhibited lymphoproliferative responses in a dose-dependent way by 50% only in healthy controls, but not in AEDS patients . Levels of inhibition of the proliferation by alpha-methylmannoside correlated inversely with the yeast- and mannan-specific IgE levels . CONCLUSIONS: These results show that in healthy subjects yeast mannan activates lymphocytes both in an HLA-DR-dependent manner and as a result of direct crosslinking of the cell surface . However, in AEDS the elevated lymphoproliferative response is HLA-DR-dependent, although only a slight proportion of this response results from direct crosslinking.

Yao Xue Xue Bao, 2001 Aug, 36(8), 590 - 4
{Chemical constituents of Diuranthera inarticulata}; Yang WL et al.; AIM: To investigate the chemical constituents from the roots of Diuranthera inarticulata Wang et K . Y . Lang . METHODS: The compounds were isolated with silica gel column chromatography and their structures were identified on the basis of spectral data and TLC with authentic samples . RESULTS: From the methanolic extracts of the roots of Diuranthera inarticulata, ten compounds were obtained and nine of them were identified as chloromaloside A (III), diuranthoside A (IV), diuranthoside B (V), diuranthoside F (VI), diuranthoside G (VII), 2 alpha, 3 alpha, 19 alpha-trihydroxy-urs-12-en-28-oic acid (VIII), 3,6,7,3',4'-pentamethoxy flavone (IX), gitogenin (X) and daucosterol (XI) . III-VI showed inhibition of growth with the yeast, Candida albicans . III, IV and VI also showed inhibition of growth with fungus, Aspergillus niger, in the test of antibacterial activity . CONCLUSION: All of these compounds were isolated from this plant for the first time . Among them, VI and VII are new steroidal saponins.

Zhongguo Zhong Xi Yi Jie He Za Zhi, 2001 Jun, 21(6), 419 - 21
{Clinical and experimental study on treatment of infantile mycotic enteritis by jiechang mixture}; Dong Y et al.; OBJECTIVE: To evaluate the therapeutic effect and mechanism of Jiechang Mixture (JCM) in treating infantile mycotic enteritis . METHODS: The children patients confirmed to be suffered from mycotic enteritis were divided randomly into two groups: the 40 cases in the treated group treated with JCM and the 20 cases in the control group treated with fungicidin . The time of stool forming, diarrhea relieving and fungi vanishing were observed . The promoting effect of JCM on small intestine motility and the inhibition of JCM on Candida albicans in stool culture were studied experimentally . RESULTS: The time of stool forming, diarrhea relieving and fungi vanishing in the treated group were significantly shorter than those in the control group (P < 0.01), and JCM also showed a better effect in improving clinical symptoms and signs of patients than the control . Results of experimental study showed that JCM could abate the ink evacuation of small intestine in mice . The fungi inhibitory test suggested that Candida albicans was susceptible to JCM . CONCLUSION: JCM is an effective herbal medicine in treating infantile mycotic enteritis.

Crit Care Med, 2003 Feb, 31(2), 501 - 7
Comparative virulence of Candida albicans yeast and filamentous forms in orally and intravenously inoculated mice; Bendel CM et al.; OBJECTIVE: Candida albicans, a dimorphic fungus that switches from yeast to filamentous forms, is a major cause of complicating systemic infection in intensive care patients . The aim of this study was to compare the pathogenic potential of C . albicans yeast and filamentous forms . DESIGN: Separate groups of mice were inoculated either intravenously or orally with C . albicans CAF2 (wild type), HLC54 (yeast forms defective in filament formation), or BCa2-10 (constitutively filamentous) . Mice were killed 1, 7, 14, and 21 days after intravenous C . albicans and kidneys and liver were quantitatively cultured; cohort groups were observed for mortality . Mice were pretreated with antibiotics for 3 days before oral inoculation with C . albicans, and killed 3 days later with dexamethasone administered for the latter 3 days; at sacrifice, the mesenteric lymph nodes and kidneys were cultured to monitor extraintestinal dissemination of C . albicans . SETTING: University teaching hospital research laboratory . SUBJECTS: Female, Swiss Webster, adult mice . MEASUREMENTS AND MAIN RESULTS: In intravenously inoculated mice, mortality was highest with wild-type C . albicans CAF2 (92%), intermediate with HLC54 (56%), and not detected with constitutively filamentous BCa2-10 (0%); BCa2-10 was cleared from the kidney and liver, but CAF2 and HLC54 were recovered at approximately 10(5-7)/g kidney and 10(4-5)/g liver . There was only occasional mortality in orally inoculated mice and the numbers of cecal C . albicans CAF2 and HLC54 were similarly high (approximately 10(7)/g), whereas numbers of cecal BCa2-10 were at least 100-fold lower . Extraintestinal dissemination was greatest with HLC54, intermediate with CAF2, and undetectable with BCa2-10 . CONCLUSIONS: Of the three C . albicans strains studied, wild-type CAF2 was most virulent in intravenously inoculated mice and HLC54 (defective in filament formation) was most virulent in orally inoculated mice . The constitutively filamentous BCa2-10 was avirulent in both models, suggesting that filamentous forms by themselves might not be critically important for C . albicans virulence.

Biol Pharm Bull, 2003 Feb, 26(2), 233 - 40
Effect of CAWS, a mannoprotein-beta-glucan complex of Candida albicans, on leukocyte, endothelial cell, and platelet functions in vitro; Kurihara K et al.; Candida albicans is a medically important fungus which induces a disseminated candidasis and candidemia in immunocompromised hosts, and releases a polysaccharide fraction into the blood . We recently found that C . albicans released a water-soluble polysaccharide fraction (CAWS) into synthetic medium and demonstrated that CAWS was mainly composed of a complex of mannan and beta-glucan . In the murine system, CAWS showed a lethality resembling anaphylactic shock when administered i.v., and induced coronary arteritis similar to Kawasaki Disease (KD) when given i.p . In the present study, we examined the biological activity of CAWS in the cell culture and found the following: i) CAWS slightly induced production of IFN-gamma and IL-6 by splenocytes at lower dose (ca . 10 micro g/ml), but at a higher dose strongly inhibited the proliferation of splenocytes induced by a B cell mitogen, lipopolysaccharide (LPS) and a T cell mitogen, concanavalin A . ii) The viability of these splenocytes monitored by propidium iodide staining was significantly reduced . iii) The addition of CAWS to a culture of monophage RAW264.7 cells significantly reduced cellular growth rate dose dependently . iv) The LPS-mediated synthesis of cytokines by RAW264.7 cells was significantly inhibited by CAWS . v) CAWS induced an aggregation of platelets in human platelet-rich plasma, and vi) CAWS inhibited the production of thrombomodulin by human umbilical endothelial cells and acted synergistically with TNF-alpha . Thus, CAWS strongly inhibited the cellular functions of leukocytes in vitro, partly through direct cytotoxicity . The enhanced production in injured cells of the vascular endothelium would be related to the local inflammatory response in the coronary artery.

Biol Pharm Bull, 2003 Feb, 26(2), 188 - 93
Synthesis and biological activities of 2,6-diaryl-3-methyl-4-piperidone derivatives; Rameshkumar N et al.; In the present study, a new series of 2,6-diaryl-3-methyl-4-piperidones was synthesized by Mannich reaction (condensation) of ethyl-methyl ketone, substituted aromatic aldehydes and ammonium acetate . Oximes and thiosemicarbazone derivatives of 2,6-diaryl-3-methyl-4-piperidones were synthesized by reaction with hydroxylamine hydrochloride and thiosemicarbazide respectively . The chemical structures were confirmed by means of IR, 1H-, 13C-NMR and mass spectral data . The compounds were screened for acute toxicity, analgesic, local anaesthetic and antifungal activity . 2-(4-Methylphenyl)-3-methyl-6-(4-chlorophenyl)-piperidin-4-one 2 exhibited the highest analgesic and local anaesthetic activity . The oximes and thiosemicarbazones were completely devoid of analgesic and local anaesthetic activity . 2-(4-Methylphenyl)-3-methyl-6-(4-hydroxyphenyl)-piperidin-4-oxime 21 and 2-(4-methoxyphenyl)-3-methyl-6-(4-chlorophenyl)-piperidin-4-oxime 17 exhibited potent antifungal activity against Aspergillus niger . Antifungal activity against Candida albicans was observed only with 2-(4-dimethylaminophenyl)-3-methyl-6-(4-chlorophenyl)-piperidin-4-oxime 20 . 2,6-Diaryl-3-methyl-4-piperidones did not exhibit antifungal property.

Microbiology, 2003 Jan, 149(Pt 1), 261 - 7
Differential expression of Candida albicans phospholipase B (PLB1) under various environmental and physiological conditions; Mukherjee PK et al.; Candida is the fourth most common organism responsible for bloodstream infections in many intensive care units, with Candida albicans being the most predominant species isolated in such cases . It has previously been shown that candidal phospholipase B, encoded by the PLB1 gene, is an important virulence factor for C . albicans pathogenesis . In this study, the effects of environmental factors (carbohydrate source and pH) and physiological conditions (serum, phospholipids and temperature) on the expression of PLB1 by C . albicans cells grown in rich {Sabouraud dextrose broth (SB) or yeast extract/peptone/dextrose} or chemically defined {Lee's, RPMI-1640 or yeast nitrogen base (YNB)} media were investigated . Northern blot analyses revealed that PLB1 mRNA was expressed in C . albicans cells grown in rich media at 30 degrees C but not at 37 degrees C . However, the protein Plb1p was detected in fungal cells growing at 37 degrees C in SB, as determined by Western blot analysis, indicating that although the mRNA for this gene was not detected, the actual gene product was present at this temperature . Expression of PLB1 was detected in cells grown in YNB/glucose at 30 degrees C but not at 37 degrees C . However, growth of C . albicans in YNB/glucose supplemented with serum and phospholipids resulted in expression of PLB1 at 37 degrees C also . Additionally, acidic pH induced higher levels of PLB1 mRNA expression compared to neutral pH, while the morphological form of C . albicans did not have any influence on the expression of this gene . The studies described here show that the expression of PLB1 is regulated by nutritional supplementation, environmental factors and the growth phase of the C . albicans cells, as well as by physiological conditions . The differential expression of PLB1 in response to environmental factors may be correlated to host-specific components available to C . albicans during infection.

Microbiology, 2003 Jan, 149(Pt 1), 249 - 59
RAM2: an essential gene in the prenylation pathway of Candida albicans; Song JL et al.; Prenylation is a post-translational protein modification process that results in correct protein localization to membranes in the cell . It is mediated by at least three prenyltransferases (PTFs) in eukaryotic cells . The RAM2 gene encodes the common alpha-subunit of two PTFs, farnesyltransferase (FTase) and geranylgeranyltransferase I (GGTase I) . In this study, the RAM2 gene of the prenylation pathway in the opportunistic pathogen Candida albicans was analysed . The heterozygote trisomy test was used to demonstrate that RAM2 is essential to the viability of C . albicans . RAM2 mRNA in the yeast and hyphae growth states was not detected by Northern blot analysis, but was detected by RT-PCR . Drugs that inhibit mammalian PTFs do not alter Candida cell growth, but they do inhibit FTase and GGTase I activities in cell-free enzyme assays . The data from genetic studies and cell-free enzyme assays suggest that the drugs may not have access to the Candida cytoplasm . The regulation of PTF subunits was also examined in a strain in which RAM2 was under the control of a regulable promoter . Overall, this study demonstrated that RAM2 is essential to C . albicans, indicating that protein prenylation is an indispensable cellular process in this yeast.

J Clin Microbiol, 2003 Feb, 41(2), 845 - 50
AFLMP1 encodes an antigenic cel wall protein in Aspergillus flavus; Woo PC et al.; We have previously reported the cloning and characterization of the MP1 gene in Penicillium marneffei and the AFMP1 gene in Aspergillus fumigatus and their use for serodiagnosis of penicilliosis and aspergilloma and invasive aspergillosis, respectively . In this study, we describe the cloning of the AFLMP1 gene, which encodes the homologous antigenic cell wall protein in Aspergillus flavus, the most common Aspergillus species associated with human disease in our locality and in other Asian countries and the second most common Aspergillus species associated with human disease in Western countries . AFLMP1 codes for a protein, Aflmp1p, of 273 amino acid residues, with a few sequence features that are present in Mp1p and Afmp1p, the homologous antigenic cell wall proteins in P . marneffei and A . fumigatus, respectively, as well as several other cell wall proteins of Saccharomyces cerevisiae and Candida albicans . It contains a serine- and threonine-rich region for O glycosylation, a signal peptide, and a putative glycosylphosphatidylinositol attachment signal sequence . Specific anti-Aflmp1p antibody was generated with recombinant Aflmp1p protein purified from Escherichia coli to allow further characterization of Aflmp1p . Indirect immunofluorescence analysis indicated that Aflmp1p is present on the surface of the hyphae of A . flavus . Finally, it was observed that patients with aspergilloma and invasive aspergillosis due to A . flavus develop a specific antibody response against Aflmp1p . This suggested that the recombinant protein and its antibody may be useful for serodiagnosis in patients with aspergilloma or invasive aspergillosis, and the protein may represent a good cell surface target for host humoral immunity.

J Clin Microbiol, 2003 Feb, 41(2), 831 - 4
Detection of Candida albicans mRNA from formalin-fixed, paraffin-embedded mouse tissues by nested reverse transcription-PCR; Schofield DA et al.; Histopathology archives represent a vast source of infectious disease specimens that can be used to elucidate important disease processes . In this report, we describe a method to detect Candida albicans gene expression from infected, formalin-fixed, paraffin-embedded mouse tissue . By use of glass beads to break open the fungal cells and proteinase K treatment, RNA was extracted routinely from tissue sections that had been fixed for up to 72 h . Upon reverse transcription of the RNA and nested PCR, the procedure detected C . albicans "housekeeping" and putative virulence genes.

J Clin Microbiol, 2003 Feb, 41(2), 712 - 6
Simple method for screening Candida species isolates for the presence of secreted proteinases: a tool for the prediction of successful inhibitory treatment; Dostal J et al.; The yeasts of the genus Candida are opportunistic pathogens associated with the rising incidence of life-threatening infections in immunocompromised individuals . Secretion of aspartic proteinases has been determined to be one of the virulence factors of the pathogenic Candida species . To analyze the extracellular proteolytic activities of a large number of Candida clinical isolates, we developed a screening system based on a solid medium containing hemoglobin as the sole nitrogen source . The cleavage of hemoglobin by the secreted proteinases results in formation of clearance zones . The visibility of such zones was enhanced by addition of an acid-base indicator . Using this system, we assessed 245 clinical isolates of Candida from patients in the hospital of the Faculty of Medicine, Palacky University, Olomouc, Czech Republic, for the presence of secreted aspartic proteases (Saps) . We also used the test plates for rapid semiquantitative testing of Sap inhibitors . Most of the pepstatin analogs affected the formation of the zones of clearance as well as the growth of Candida albicans, C . tropicalis, and C . parapsilosis colonies . By contrast, the human immunodeficiency virus proteinase inhibitors saquinavir, ritonavir, nelfinavir, and indinavir had no effect on the Candida strains tested . These results are in agreement with the inhibition constants obtained for the individual inhibitors with purified Saps . Thus, the plates containing hemoglobin proved to be an appropriate tool for the rapid and reliable assessment of Sap production and inhibition.

J Clin Microbiol, 2003 Feb, 41(2), 581 - 5
Detection and identification of fungi from fungus balls of the maxillary sinus by molecular techniques; Willinger B et al.; The aim of this study was to find a reliable method for the detection and identification of fungi in fungus balls of the maxillary sinus and to evaluate the spectrum of fungi in these samples . One hundred twelve samples were obtained from patients with histologically proven fungal infections; 81 samples were paraffin-embedded tissue sections of the maxillary sinus . In 31 cases, sinus contents without paraffin embedding were sent for investigation . PCR amplification with universal fungal primers for 28S ribosomal DNA and amplicon identification by hybridization with species-specific probes for Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus, Aspergillus glaucus, Pseudallescheria boydii, Candida albicans, and Candida glabrata were performed for all samples . Furthermore, PCR products were sequenced . Fresh samples were also cultivated . Fungal DNA was detected in all of the fresh samples but only in 71 paraffin-embedded tissue samples (87.7%) . Sequence analysis was the most sensitive technique, as results could be obtained for 28 (90.3%) fresh samples by this method in comparison to 24 (77.4%) samples by hybridization and 16 (51.6%) samples by culture . However, sequence analysis delivered a result for only 36 (50.7%) of the paraffin-embedded specimens . Hybridization showed reliable results for A . fumigatus, which proved to be the most common agent in fungus balls of the maxillary sinus . Other Aspergillus species and other genera were rarely found.

J Clin Microbiol, 2003 Feb, 41(2), 552 - 7
Highly polymorphic microsatellite for identification of Candida albicans strains; Sampaio P et al.; The polymorphism of a new microsatellite locus (CAI) was investigated in a total of 114 Candida albicans strains, including 73 independent clinical isolates, multiple isolates from the same patient, isolates from several episodes of recurrent vulvovaginal infections, and two reference strains . PCR genotyping was performed automatically, using a fluorescence-labeled primer, and in the 73 independent isolates, 26 alleles and 44 different genotypes were identified, resulting in a discriminatory power of 0.97 . CAI was revealed to be species specific and showed a low mutation rate, since no amplification product was obtained when testing other pathogenic Candida species and no genotype differences were observed when testing over 300 generations . When applying this microsatellite to the identification of strains isolated from recurrent vulvovaginal infections in eight patients, it was found that 13 out of 15 episodes were due to the same strain . When multiple isolates, obtained from the same patient and plated simultaneously, were typed for CAI, the same genotype was found in each case, confirming that the infecting population was clonal . Moreover, the same genotype appeared in isolates from the rectum and the vagina, revealing that the former could be a reservoir of potentially pathogenic strains . This new microsatellite proves to be a valuable tool to differentiate C . albicans strains . Furthermore, when compared to other molecular genotyping techniques, CAI proved to be very simple, highly efficient, and reproducible, being suitable for low-quantity and very-degraded samples and for application in large-scale epidemiological studies.

Mycoses, 2002 Oct, 45(8), 306 - 12
Prevalence of Candida spp . in hospitalized patients and their risk factors; Resende JC et al.; A total of 133 Candida spp . strains originating from a group of 100 patients from Santa Casa de Misericordia of Belo Horizonte, Brazil, between March 1995 and December 1996, were first identified and classified into six different species: Candida albicans (51%), C . tropicalis (33%), C . parapsilosis (8%), C . glabrata (5%), C . krusei (2%) and C . guilliermondii (1%) . All C . albicans strains were serotyped and 55% of these were found to belong to serotype A and 45% belonging to serotype B . The medical records of each patient were examined to characterise and survey the main risk factors associated with them . Most of the patients were between 60- and 80-years-old, 53% were males and 47% were females . Most patients were from the intensive care unit (ICU) . Only 10 patients were not exposed to antimicrobial agents and 72 patients were not prescribed antifungal agents . Forty patients showed no other clinical condition and all showed some underlying disease that justified hospitalization . Eighty-seven patients had undergone some invasive procedure and 31 patients had been submitted to two different procedures simultaneously.

Mycoses, 2002 Oct, 45(8), 300 - 5
Impact of components of denture acrylic resin on gingival cell growth and sensitivity to Candida albicans adhesion; Makihira S et al.; The effects of four liquid components of denture acrylic resin on host cell activity and fungal adhesion were investigated in this study . The low concentration (1 micromol l(-1)) of the liquid components caused no change in the activities and morphologies of the gingival fibroblast cells, compared with control and dimethylsulphoxide-exposed cells . However, when the cells were exposed to high concentrations (1 mmol l(-1)) of benzqyl peroxide, morphological change was observed, implying that the exposure of the cells to high concentrations of the liquid components of denture acrylic causes the loss of adhesion proteins from the cells . Thus the amount of Candida adhesion to human gingival cells was analysed, and the adherence of fungi to the cell was significantly reduced when the cells were pre-exposed to methyl methacrylate, hydroquinone and benzoyl peroxide at a concentration of 1 micromol l(-1) (P < 0.01), which did not affect either the cell viability or the cell morphology . These results, taken together, suggested that the renewal of dentures could be a possible therapeutic and/or preventive aid for oral candidosis in denture-wearing patients.

Mycoses, 2002 Oct, 45(8), 253 - 8
Candida albicans and the principle of opportunism . an essay; Niewerth M et al.; The terms 'opportunism' and 'opportunist' have been used for many years in various fields encompassing politics, social science, economics, and life sciences . However these terms are still ill-defined . The inter-relationship between the various meanings in different given contexts is discussed under the special aspect of micro-organisms, and in particular Candida albicans, which together with Pseudomonas aeruginosa, is most often addressed as an opportunist or suspected of opportunism.

Biometals, 2003 Jun, 16(2), 321 - 9
Mode of anti-fungal activity of 1,10-phenanthroline and its Cu(II), Mn(II) and Ag(I) complexes; Coyle B et al.; The mode of action of the anti-fungal compounds, 1,10-phenanthroline (phen), {Cu(phen)2(mal)} x 2H2O, {Mn(phen)2(mal)} x 2H2O and {Ag2(phen)3(mal)} x 2H2O (malH2 = malonic acid), was examined using the pathogenic yeast Candida albicans . The compounds have minimum inhibitory concentrations (MIC's) in the range 1.25-5.0 microg cm(-3) and at a concentration of 10 microg cm(-3) display some fungicidal activity . Yeast cells exposed to these drugs show a diminished ability to reduce 2,3,5-triphenyltetrazolium chloride (TTC), indicating a reduction in respiratory function . Treating exponential and stationary phase yeast cells with phen and the Cu(II) and Mn(II) complexes causes a dramatic increase in oxygen consumption . All of the drugs promote reductions in the levels of cytochromes b and c in the cells, whilst the Ag(I) complex also lowers the amount of cytochrome aa3 . Cells treated with phen and the Cu(II) and Ag(I) species show reduced levels of ergosterol whilst the Mn(II) complex induces an increase in the sterol concentration . The general conclusion is that the drugs damage mitochondrial function and uncouple respiration . That the drugs are not uniformly active suggests their bioactivity has a degree of metal-ion dependency . Phen and metal-phen complexes represent a novel set of highly active anti-fungal agents whose mode of action is significantly different to that of the polyene and azole prescription drugs.

J Med Chem, 2003 Feb 13, 46(4), 474 - 85
Structure-based de novo design, synthesis, and biological evaluation of non-azole inhibitors specific for lanosterol 14alpha-demethylase of fungi; Ji H et al.; The active site of lanosterol 14alpha-demethylase (CYP51) was investigated via MCSS functional group mapping and LUDI calculations . Several non-azole lead molecules were obtained by coupling structure-based de novo design with chemical synthesis and biological evaluation . All of the lead molecules exhibited a strong inhibitory effect on CYP51 of Candida albicans . They occupy the substrate-binding site and interfere with the binding of azole antifungal agents in a competitive manner . The mode of action of the lead molecules was validated by spectrophotomeric analysis and SAR studies . This is the first successful example reported for the inhibitor design of the cytochrome P450 superfamily using the de novo design strategy . Because the affinity of the lead molecules for CYP51 was mainly attributed to their nonbonding interaction with the apoprotein, the studies presented here afford the opportunity to develop novel antifungal agents that specifically interact with the residues in the active site and avoid the serious toxicity arising from coordination binding with the heme of mammalian P450s.

Bioorg Med Chem Lett, 2003 Feb 10, 13(3), 495 - 7
In vivo characterization of A-192411: a novel fungicidal lipopeptide (II); Meulbroek JA et al.; The ability of the novel antifungal cyclic hexalipopetide A-192411 to treat fungal infections in rodents is presented . Efficacy was demonstrated against Candida albicans as both prolonged survival of systemically infected mice and clearance of viable yeasts from kidneys . The efficacy of A-192411, administered intraperitoneally, was equivalent to amphotercin B at a 4-fold lower dose by weight in the systemic candidiasis models in mice, while the efficacy of A-192441 administered intravenously was equivalent to amphotercin B by weight in the Candida pyelonephritis model in rats . A-192411 also slightly prolonged the survival of immunocompromised mice infected systemically with Aspergillus fumigatus.

Bioorg Med Chem Lett, 2003 Feb 10, 13(3), 433 - 6
Novel antifungal beta-amino acids: synthesis and activity against Candida albicans; Mittendorf J et al.; A series of novel beta-amino acids has been synthesized and tested for their in vitro antifungal activity against Candida albicans . A steep SAR was observed . beta-Amino acid 21 (BAY 10-8888/PLD-118) revealed the most favourable activity-tolerability profile and was selected for clinical studies as a novel antifungal for the oral treatment of yeast infections.

Biochem Biophys Res Commun, 2003 Feb 7, 301(2), 529 - 34
The signal peptide NPFSD fused to ricin A chain enhances cell uptake and cytotoxicity in Candida albicans; Rajarao GK et al.; Microorganisms possess stringent cell membranes which limit the cellular uptake of antimicrobials . One strategy to overcome these barriers is to attach drugs or research reagents to carrier peptides that enter cells by passive permeation or active uptake . Here the short endocytosis signal peptide NPFSD was found to efficiently deliver both FITC and GFP into Saccharomyces cerevisiae and Candida albicans with uptake into the majority of cells in a population . The NPFSD signal is itself non-toxic, but when fused to the ricin A chain toxin (RTA) the peptide enhanced both cell uptake and toxicity against C . albicans, which like other yeasts is resistant to naked RTA . Cell entry required at least 1 h incubation, temperatures above 4 degrees C, and an energy source, and uptake was out-competed with free peptide . Therefore, the NPFSD peptide can carry a range of compounds into yeasts and this delivery route holds promise to enhance the activity of antifungals.

Farmaco, 2002 Dec, 57(12), 973 - 7
Synthesis antimicrobial and antifungal activity of some new 3 substituted derivatives of 4-(2,4-dichlorophenyl)-5-adamantyl-1H-1,2,4-triazole; Papakonstantinou-Garoufalias S et al.; The synthesis of a series of substituted hydrazones and thiazolidinones is described, starting from N-{4-(2,4-dichlorophenyl)-5-adamantyl-1H-1,2,4-triazol-3-ylmercaptoacetyl}hydrazine . The new compounds were tested for antimicrobial and antifungal activity and some of them exhibited moderate activity against Candida albicans.

J Antimicrob Chemother, 2003 Feb, 51(2), 305 - 12
The activity of amphotericin B against Candida albicans is not directly associated with extracellular calcium concentration; Rogers PD et al.; The ability of amphotericin B to increase intracellular calcium concentrations in human cells is associated with the toxicity of this antifungal agent . The present study was performed to determine whether amphotericin B affects the influx or efflux of calcium in Candida albicans, and whether the antifungal activity of amphotericin B is dependent upon extracellular calcium concentrations . Concentration-response studies demonstrated that the addition of up to 1 mM EGTA to standard growth medium, with a more than 4000-fold decrease in extracellular calcium concentration, had no effect on the activity of amphotericin B against C . albicans . Amphotericin B did affect the kinetics of calcium influx acutely (< or =10 min), but had no net effect on long-term (1-24 h) calcium accumulation . Calcium efflux was also not affected by amphotericin B . These results indicate that, unlike its effects on mammalian cells, the toxicity of amphotericin B against C . albicans is not dependent upon increased movement of calcium across the cell membrane or the presence of extracellular calcium.

J Antimicrob Chemother, 2003 Feb, 51(2), 281 - 7
Treatment of plastic and extracellular matrix components with chlorhexidine or benzalkonium chloride: effect on Candida albicans adherence capacity in vitro; Imbert C et al.; This study investigates the influence of treatment of plastic and extracellular matrix (ECM) proteins with chlorhexidine or benzalkonium chloride on subsequent adherence of Candida albicans . Three concentrations were tested for each antiseptic: (i) chlorhexidine, MIC (6.25-12.5 mg/L), 80 x MIC and 800 x MIC; and (ii) benzalkonium chloride, MIC (3.12 mg/L), 40 x MIC and 1600 x MIC . Chlorhexidine and benzalkonium chloride activities were correlated with the tested concentrations . Antiseptics used at MIC were unable to modify the adherence to plastic or ECM proteins . Chlorhexidine (80 x MIC) induced a decrease in plastic adherence of 31% of the 15 strains used and an increase in ECM protein adherence of 13% of strains . Benzalkonium chloride (40 x MIC) induced a decrease in adherence to ECM proteins or plastic of 13-27% of strains . Our results indicated that the treatment with 1600 x MIC benzalkonium chloride could induce the opposite effect on adherence, depending on the surface: 60% of the strains showed an increase in their adherence to ECM proteins, whereas 93% of the strains showed a decrease in their adherence to plastic . A similar phenomenon was observed after treatment with 800 x MIC chlorhexidine: 60% of the strains showed an increase in their adherence to ECM proteins, whereas 67% showed a decrease in adherence to plastic . Treatment of medical devices with at least 5000 mg/L of chlorhexidine or benzalkonium chloride could therefore reduce C . albicans adherence to plastic surfaces, but would be unable to prevent fungal adherence to ECM proteins.

Fungal Genet Biol, 2003 Feb, 38(1), 133 - 41
The two isoforms of the cAMP-dependent protein kinase catalytic subunit are involved in the control of dimorphism in the human fungal pathogen Candida albicans; Cloutier M et al.; We have cloned the Candida albicans TPK2 gene encoding a cAMP-dependent protein kinase (PKA) catalytic subunit and generated a tpk2 homozygous null mutant to assess its ability to germinate in liquid media . N-acetylglucosamine (GlcNAc)-induced germ-tube formation was attenuated in the tpk2 strain and enhanced by compounds that are known to increase the PKA activity in situ . Germination was completely blocked in the presence of the myristoylated derivative of the heat-stable PKA inhibitor (MyrPKI) . These results indicate that TPK1 acts positively in regulating the morphogenetic transition in C . albicans in the absence of the TPK2 gene . We were able to identify an mRNA from this second form of PKA in both wild-type and tpk2 null mutant cells . We found that PKA activity measured in the mutant lacking the TPK2 gene was about 10% of that displayed by the wild-type . The finding that the germinative response of tpk2 null mutant to serum was severely diminished at low serum concentrations indicates that the level of PKA is an important determinant of filamentous growth at low serum concentrations . The extent of germination attained at higher serum concentrations (5%) was similar in the wild-type and in the tpk2 null mutant strains suggesting that under these conditions germination was triggered through a PKA-independent pathway.

Chang Gung Med J, 2002 Nov, 25(11), 778 - 82
Endogenous Candida endophthalmitis after two consecutive procedures of suction dilatation and curettage; Chang TS et al.; Endogenous Candida endophthalmitis (ECE) is a rare disease . We present a patient with Candida endophthalmitis after two consecutive procedures of suction dilatation and curettage for elective abortion . A 24-year-old single woman who received a suction dilatation and curettage one week ago developed pain and blurred vision in the right eye . Endogenous Candida endophthalmitis was diagnosed and treated with oral fluconazole and pars plana vitrectomy with adjunction of intravitreal amphotericin B injection . The vitreous culture revealed Candida albicans . The vitreous inflammation subsided greatly after the initial treatment but flared up after the second dilatation and curettage for incomplete abortion 5 days after the vitrectomy . The oral fluconazole was replaced by intravenous amphotericin B, and a second vitrectomy with injection of intravitreal amphotericin B was performed . Postoperatively, the intraocular inflammation resolved gradually . Six months after the second vitrectomy, the best-corrected visual acuity in the right eye was 20/25 . The excellent visual acuity of this patient was attributed to the early diagnosis and aggressive treatment . For patients with mild disease, less toxic oral fluconazole as the systemic antifungal agent instead of more toxic intravenous amphotericin B has been recommended . For those with advanced disease, intravitreal amphotericin B in conjunction with vitrectomy has been advocated by many eye surgeons.

Unfallchirurg, 2003 Jan, 106(1), 70 - 2
{Candida infection in hip alloarthroplasty}; Prenzel KL et al.; A 46-year-old polytoxicomanic patient underwent a total hip replacement for necrosis of the femoral head . The patient suffered additionally from chronic pancreatitis with insulin-dependent diabetes and polyneuropathy . Three weeks later he developed a deep wound infection followed by surgical revision . The infection persisted despite further revision operations and systemically and locally applied antibiotics . After removal of the prosthesis,microbiology revealed Staphylococcus aureus, enterococcus,and Candida parapsilosis . Five additional revision operations, application of suction-irrigation drainage, and systemically administered antibiotics could not stop the infectious process . The patient was transferred to our institution 3 months after primary surgery . The joint defect was filled with a PMMA Palacos spacer . This time biopsies were only positive for Candida albicans . After 12 days of antifungal therapy with fluconazole,microbiological biopsies were sterile . The spacer was removed and femoral extension applied . After 4 weeks of further antifungal therapy, a revision prosthesis was implanted . Until now no signs of infection have appeared.

Proc Natl Acad Sci U S A, 2003 Feb 18, 100(4), 1530 - 4 Epub 2003 Jan 27.
Uptake and antifungal activity of oligonucleotides in Candida albicans; Disney MD et al.; Candida albicans is a significant cause of disease in immunocompromised humans . Because the number of people infected by fungal pathogens is increasing, strategies are being developed to target RNAs in fungi . This work shows that oligonucleotides can serve as therapeutics against C . albicans . In particular, oligonucleotides are taken up from cell culture medium in an energy-dependent process . After uptake, oligonucleotides, including RNA, remain mostly intact after 12 h in culture . For culture conditions designed for mammalian cells, intracellular concentrations of oligonucleotides in C . albicans exceed those in COS-7 mammalian cells, suggesting that uptake can provide selective targeting of fungi over human cells . A 19-mer 2'OMe (oligonucleotide with a 2'-O-methyl backbone) hairpin is described that inhibits growth of a C . albicans strain at pH < 4.0 . This pH is easily tolerated in some parts of the body subject to C . albicans infections . In vivo dimethyl sulfate modification of ribosomal RNA and the decreased rate of protein synthesis suggest that this hairpin's activity may be due to targeting the ribosome in a way that does not depend on base pairing . Addition of anti-C . albicans oligonucleotides to COS-7 mammalian cells has no effect on cell growth . Evidently, oligonucleotides can selectively serve as therapeutics toward C . albicans and, presumably, other pathogens . Information from genome sequencing and functional genomics studies on C . albicans and other pathogens should allow rapid design and testing of other approaches for oligonucleotide therapies.

J Biol Chem, 2003 Apr 11, 278(15), 13086 - 93 Epub 2003 Jan 27.
Candida albicans phospholipomannan promotes survival of phagocytosed yeasts through modulation of bad phosphorylation and macrophage apoptosis; Ibata-Ombetta S et al.; The surface of the pathogenic yeast Candida albicans is coated with phospholipomannan (PLM), a phylogenetically unique glycolipid composed of beta-1,2-oligomannosides and phytoceramide . This study compared the specific contribution of PLM to the modulation of signaling pathways linked to the survival of C . albicans in macrophages in contrast to Saccharomyces cerevisiae . C . albicans endocytosis by J774 and disregulation of the ERK1/2 signal transduction pathway was associated downstream with a reduction in Bad Ser-112 phosphorylation and disappearance of free Bcl-2 . This suggested an apoptotic effect, which was confirmed by staining of phosphatidylserine in the macrophage outer membrane . The addition of PLM to macrophages incubated with S . cerevisiae mimicked each of the disregulation steps observed with C . albicans and promoted the survival of S . cerevisiae . Externalization of membranous phosphatidylserine, loss of mitochondrial integrity, and DNA fragmentation induced by PLM showed that this molecule promoted yeast survival by inducing host cell death . These findings suggest strongly that PLM is a virulence attribute of C . albicans and that elucidation of the relationship between structure and apoptotic activity is an innovative field of research.

Shi Yan Sheng Wu Xue Bao, 2001 Mar, 34(1), 25 - 33
{Knocking out of CEK2 and CSK1 affect the phenotype of Candida albicans}; Chen J et al.; We have cloned two novel mitogen-activated protein kinase (MAPK) genes CEK2 and CSK1 from Candida albicans strain SC5314 . The heterozygous and homozygous cek 2 and csk 1 mutants were obtained by homologous recombination . The cek2 and csk1 null mutant strains CAJ3 and CAJ6 grew as wrinkled clonis on solid YPD medium . Expression of CEK2 gene in strain CAF3-1 repressed the filamentous growth of the strain weakly . Two-hybrid results showed Cek2 and Csk1 could not interact with Cph1 and Hst7 directly . Cek2 and Csk1 might cooperate with the filamentous transcriptional repressor Tup1.

Mol Cell Proteomics, 2002 Dec, 1(12), 967 - 82
Sequential fractionation and two-dimensional gel analysis unravels the complexity of the dimorphic fungus Candida albicans cell wall proteome; Pitarch A et al.; The cell wall proteins of Candida albicans play a key role in morphogenesis and pathogenesis and might be potential target sites for new specific antifungal drugs . However, these proteins are difficult to analyze because of their high heterogeneity, interconnections with wall polysaccharides (mannan, glucan, and chitin), low abundance, low solubility, and hydrophobic nature . Here we report a subproteomic approach for the study of the cell wall proteins (CWPs) from C . albicans yeast and hyphal forms . Most of the mannoproteins present in this compartment were extracted by cell wall fractionation according to the type of interactions that they establish with other structural components . CWPs were solubilized from isolated cell walls by hot SDS and dithiothreitol treatment followed by extraction either by mild alkali conditions or by enzymatic treatment with glucanases and chitinases . These highly enriched cell wall fractions were analyzed by two-dimensional PAGE, showing that a large number of proteins are involved in cell wall construction and that the wall remodeling that occurs during germ tube formation is related to changes in the composition of CWPs . We suggest that the CWP-chitin linkage is an important retention mechanism of CWPs in C . albicans mycelial forms . This article also highlights the usefulness of the combination of sequential fractionation and two-dimensional PAGE followed by Western blotting using specific antibodies against known CWPs in the characterization of incorporation mechanisms of such CWPs into the cell wall and of their interactions with other wall components . Mass spectrometry analyses have allowed the identification of several cell surface proteins classically associated with both the cell wall and other compartments . The physiological significance of the dual location of these moonlighting proteins is also discussed . This approach is therefore a powerful tool for obtaining a comprehensive and integrated view of the cell wall proteome.

Antimicrob Agents Chemother, 2003 Feb, 47(2), 601 - 6
Efficacy of CS-758, a novel triazole, against experimental fluconazole-resistant oropharyngeal candidiasis in mice; Kamai Y et al.; The therapeutic efficacy of CS-758, a novel triazole, was evaluated against experimental murine oropharyngeal candidiasis induced by Candida albicans with various susceptibilities to fluconazole . Against infections induced by strains with various susceptibilities to fluconazole, the efficacy of fluconazole was strongly correlated with the MIC of fluconazole, as measured by the NCCLS method, and agreed with the NCCLS interpretive breakpoints, suggesting that the efficacies of new drugs could be predicted by using this model . The results of the fungal burden study corresponded with the results of the histopathological study . CS-758 exhibited potent in vitro activity (MICs, 0.004 to 0.06 micro g/ml) against the strains used in this murine model including fluconazole-susceptible dose-dependent and fluconazole-resistant strains (fluconazole MICs, 16 to 64 micro g/ml) . CS-758 exhibited excellent efficacy against the infections induced by all the strains including a fluconazole-resistant strain, and the reductions in viable cell counts were significant at 10 and 50 mg/kg of body weight/dose . Fluconazole was not effective even at 50 mg/kg/dose against infections induced by a fluconazole-resistant strain (fluconazole MIC, 64 micro g/ml) . These results suggest that CS-758 is a promising compound for the treatment of oropharyngeal candidiasis including fluconazole-refractory infections.

Diagn Microbiol Infect Dis, 2002 Dec, 44(4), 353 - 7
Polymicrobial candidemia; Pulimood S et al.; We conducted a retrospective review of the clinical data on patients with polymicrobial candidemia during a nine-year period (1992-2000) at our tertiary care University Hospital . Also, the clinical features of patients with polymicrobial vs monomicrobial candidemia were compared . There were 16 (5.2%) patients with multiple candidal species in blood among 303 patients with candidemia . Patients' age varied from 21-85 years (median 52 years); they had serious underlying co-morbidities with prolonged hospitalization (median duration 13 days); all had intravenous vascular catheters, had been exposed to multiple antibiotics and were heavily colonized with Candida . Concomitant serious bacterial infections were common (56%) . Candida albicans was isolated from blood in 11 of 16 patients; other species were C . glabrata (7 patients), C . tropicalis (6 patients) and C . parapsilosis (4 patients) . Sixty seven percent (8 of 12) patients had causative Candida species isolated from vascular catheter tip culture . Polymicrobial candidemia occurred in sicker, non-oncologic patients with frequent concomitant bacterial infections, as compared to those with monomicrobial candidemia . Crude mortality was 43%, a rate similar to that seen with monomicrobial candidemia . Polymicrobial candidemia is uncommon, seen in hospitalized patients with multiple co-morbidities and heavy candidal colonization; removal of vascular catheter and institution of antifungal therapy are important therapeutic maneuvers.

J Nat Prod, 2003 Jan, 66(1), 51 - 6
Melophlins C-O, thirteen novel tetramic acids from the marine sponge Melophlus sarassinorum; Wang CY et al.; Thirteen new metabolites, melophlins C-O (1-13), were identified from the marine sponge Melophlus sarassinorum . Compounds 1-13 represent tetramic acid derivatives that differ with regard to the nature of their alkyl side chains . The structures of the new compounds were elucidated on the basis of comprehensive spectral analysis (1H, 13C, 1H-1H COSY, HMQC, and HMBC NMR, as well as low- and high-resolution ESIMS and EIMS) . The absolute configurations of 1, 8, 10, 11, 12, and 13 were determined by ESI LC/MS analysis of chiral derivatives obtained upon oxidation and hydrolysis of the respective parent compounds . Melophlin C (1) displayed pronounced antibacterial activity against Bacillus subtilisand Staphylococcus aureus, together with antifungal activity against Candida albicans.

Lin Chuang Er Bi Yan Hou Ke Za Zhi, 2001 Feb, 15(2), 77 - 9
{Study on susceptibility test of pathogenic fungi from otorhinolaryngology by Etest method}; Li J et al.; OBJECTIVE: To study the sensitivity of pathogenic fungi from otorhinolaryngology to 5 antifungal agents of Ketoconazole (KE), Itraconazole(IT), Fluorocytosine(FC), Amphtericin B (AP) and Fluconazole(FL) by Etest method . METHOD: Etest for determine MIC of 14 pathogenic fungi was performed according to the manufacturer's instructions . The Etest strip containing the 5 antifungal drugs were putted on medium with Candida albicans, A . flarus and A . fumigatus etc., respectively, to determine MIC of antifungal drugs . RESULT: Candida albicans were sensitized to all the above 5 antifungal drugs, KE and IT were most sensitized . Seven fungi such as A . flavus, A fumigatus, A . oryzae etc were all sensitized to KE and IT . The MIC range of KE and IT against 15 strains of pathogenic fungi were < or = 0.008-2 mg/L and < or = 0.006-4 mg/L respectively . In 15 strains, 11 strains showed drug-resistant to FL, 8 strains to AP and FC, 4 strains to KE and 1 strains to IT respectively and MIC were all > 32 mg/L . CONCLUSION: Etest MIC were in good agreement with macrobroth MIC . The use of Etest strips for antifungal susceptibility test is a new and promising method with advantage of easy to perform, exact results and satisfactory reproducibility . Etest is valuable on clinical practice.

Lin Chuang Er Bi Yan Hou Ke Za Zhi, 1999 Oct, 13(10), 438 - 40
{The observation of mycology and clinical efficacy in 325 cases with otomycosis}; Nong H et al.; OBJECTIVE: To study otomycosis of pathogenic fungi, antifungal susceptibility test and clinical efficacy . METHOD: The study was performed as follows: 1) culture of pathogenic fungi and fungus identification in 110 cases . 2) antifungal susceptibility test with Etest strip by ketoconazole (KE), itraconazole (IT), Fluconazole (FC), Fluorocytosine (FL), and amphotericinum B (AP) . 3) Topical application of doktarin ps cream (DC), ketoconazole cream (KEC), clotrimazole ung (CU), and thymol alcohol (TA) were performed in 4 groups and the intermittent impulsive treatment in who failed to respond to local treatment were performed too . RESULT AND CONCLUSION: 1) Chiefly, otomycosis of pathogenic fungi were aspergillus (79%) and candida albicans (8%) and penicillium citrinum (5%) . 2) Antifungal susceptibility test showed that aspergillus were sensitive to KE and IT, but not sensitive to KE and FL, and candida albicans were sensitive to KE, IT, FC, FL and AP all; P . citrinum was sensitive to AP, FL and KE but not to FC . 3) Effective rate of topical application on 4 groups were 97.6%, 97.5%, 90.0% and 80.0% respectively . Comparing the results between compound azoles drugs and TA, there was a statistical significant difference (P < 0.05), the efficacy of compound azoles is better than that of TA . The effective rate of systematic intermittent impulsive treatment in 30 cases failed to respond to local treatment were 100%.

Infect Immun, 2003 Feb, 71(2), 956 - 63
Immunohistochemical evaluation of T cells in oral lesions from human immunodeficiency virus-positive persons with oropharyngeal candidiasis; Myers TA et al.; Oropharyngeal candidiasis (OPC), caused by Candida albicans, is the most frequent opportunistic fungal infection in human immunodeficiency virus (HIV)-positive persons . Although Th1-type CD4(+) T cells are considered important for host defense against mucosal C . albicans infections, there is a paucity of information regarding the presence and/or role of T cells in OPC lesions . In pursuit of this, initial chromophore immunohistochemical studies showed a majority of CD8(+) rather than CD4(+) cells equally distributed throughout the buccal mucosa of OPC(-) persons (HIV(-) or HIV(+)), irrespective of blood CD4(+) cell numbers . In contrast, CD8(+) cells in lesions from HIV(+) OPC(+) persons were in significantly higher numbers and concentrated at the lamina propria-epithelium interface, a considerable distance from the Candida at the outer epithelium . Dual fluorescence and confocal microscopy confirmed that the majority of CD8(+), but not CD4(+), cells were T cells by the presence or absence, respectively, of CD3 on each cell type . These results suggest that CD8(+) T cells may be important for oral host defense against OPC, especially when CD4 cell numbers are reduced, with a potential CD8 cell-specific dysfunction associated with susceptibility to OPC.

Hua Xi Kou Qiang Yi Xue Za Zhi, 2001 Oct, 19(5), 281 - 2
{Phospholipase activities of Candida albicans isolates from oral pre-malignant lesions and oral cancers}; Zhou M et al.; OBJECTIVE: The aim of this study is to compare activities of phospholipase of Candida albicans isolates from oral cavities of two groups of patients, including oral pre-malignant lesions and oral cancers . METHODS: A total of 55 isolates, including oral pre-malignant lesions (21), oral cancers (15), and normal health control (19), were studied using the precipitate zone assay on the egg yolk agar . RESULTS: No significant difference was found in phospholipase activities between these isolates from normal controls and oral pre-malignant lesions, while the activities of phospholipase of isolates from oral cancers were higher than those of the normal controls . CONCLUSION: Candida albicans may play different roles in different manners in these two closely related diseases.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2003 Jan, 95(1), 51 - 9
Reassessing the presence of Candida albicans in denture-related stomatitis; Barbeau J et al.; OBJECTIVE: The aim of this study was to reevaluate the link between Candida albicans and denture-related stomatitis according to a modified Newton classification, which reflects the classic types of inflammation as well as the extent to which the tissue is affected . STUDY DESIGN: Two groups of denture wearers were evaluated for denture-related stomatitis . The presence and number of yeasts on the dentures, the identification of the Candida species present, and the amount of plaque coverage were determined . Putative risk factors were included in the study . Relations between these variables and stomatitis were analyzed statistically . RESULTS: According to Newton's classification, the presence of yeast on the denture was not linked to whether subjects had stomatitis . But with our classification, higher prevalence of yeast carriers, yeast colony number, and plaque coverage were found on the dentures of individuals with the most extensive inflammation, regardless of Newton type . Among risk factors evaluated, wearing dentures at night and smoking were associated with the most extensive inflammation . We also demonstrated that the presence of C albicans as well as the cohabitation of different Candida species was more frequent in denture-related stomatitis . The differences were statistically significant . CONCLUSIONS: Statistical analysis of microbiologic data from different denture-related stomatitis categories according to our modified classification showed that the presence of yeast on dentures was significantly associated with the extent of the inflammation, rather than with the Newton type . Our findings suggest that the inflammatory process of stomatitis favors the colonization of Candida . These results could have new implications for diagnosis and management of the condition.

Pediatr Res, 2003 Feb, 53(2), 211 - 6
Antimicrobial polypeptides of human vernix caseosa and amniotic fluid: implications for newborn innate defense; Yoshio H et al.; Antimicrobial peptides/proteins are widespread in nature and play a critical role in host defense . To investigate whether these components contribute to surface protection of newborns at birth, we have characterized antimicrobial polypeptides in vernix caseosa (vernix) and amniotic fluid (AF) . Concentrated peptide/protein extracts were obtained from 11 samples of vernix and six samples of AF and analyzed for antimicrobial activity using an inhibition zone assay . Proteins/peptides in all vernix extracts exhibited strong antibacterial activity against Bacillus megaterium (strain Bm11), in addition to antifungal activity against Candida albicans, whereas AF-derived proteins/peptides showed only the former activity . Fractions obtained after separation by reverse-phase HPLC exhibited antibacterial activity, with the most pronounced activity in a fraction containing alpha-defensins (HNP1-3) . The presence of HNP1-3 was proved by dot blot analysis and confirmed by mass spectrometry . Lysozyme and ubiquitin were identified by sequence analysis in two fractions with antibacterial activity . Fractions of vernix and AF were also positive for LL-37 with dot blot and Western blot analyses, and one fraction apparently contained an extended form of LL-37 . Interestingly, psoriasin, a calcium-binding protein that is up-regulated in psoriatic skin and was found recently to exhibit antimicrobial activity, was characterized in the vernix extract . The presence of all of these antimicrobial polypeptides in vernix suggests that they are important for surface defense and may have an active biologic role against microbial invasion at birth.

J Immunol, 2003 Feb 1, 170(3), 1452 - 61
CCL28 has dual roles in mucosal immunity as a chemokine with broad-spectrum antimicrobial activity; Hieshima K et al.; CCL28 is a CC chemokine signaling via CCR10 and CCR3 that is selectively expressed in certain mucosal tissues such as exocrine glands, trachea, and colon . Notably, these tissues commonly secrete low-salt fluids . RT-PCR analysis demonstrated that salivary glands expressed CCL28 mRNA at the highest levels among various mouse tissues . Single cells prepared from mouse parotid glands indeed contained a major fraction of CD3(-)B220(low) cells that expressed CCR10 at high levels and CCR3 at low levels and responded to CCL28 in chemotaxis assays . Morphologically, these cells are typical plasma cells . By immunohistochemistry, acinar epithelial cells in human and mouse salivary glands were strongly positive for CCL28 . Furthermore, human saliva and milk were found to contain CCL28 at high concentrations . Moreover, the C terminus of human CCL28 has a significant sequence similarity to histatin-5, a histidine-rich candidacidal peptide in human saliva . Subsequently, we demonstrated that human and mouse CCL28 had a potent antimicrobial activity against Candida albicans, Gram-negative bacteria, and Gram-positive bacteria . The C-terminal 28-aa peptide of human CCL28 also displayed a selective candidacidal activity . In contrast, CCL27, which is most similar to CCL28 and shares CCR10, showed no such potent antimicrobial activity . Like most other antimicrobial peptides, CCL28 exerted its antimicrobial activity in low-salt conditions and rapidly induced membrane permeability in target microbes . Collectively, CCL28 may play dual roles in mucosal immunity as a chemoattractant for cells expressing CCR10 and/or CCR3 such as plasma cells and also as a broad-spectrum antimicrobial protein secreted into low-salt body fluids.

Infect Dis Obstet Gynecol, 2002, 10(2), 89 - 92
Can the diagnosis of recurrent vulvovaginal candidosis be improved by use of vaginal lavage samples and cultures on chromogenic agar?
Novikova N, Rodrigues A, Mardh PA.
OBJECTIVE: To investigate if introital and vaginal flushing samples inoculated on chromogenic agar could increase the recovery rate and rapid identification of Candida and non-albicans species, as compared to culture of posterior vaginal fornix samples on Sabouraud agar and speciation of isolates by biochemical tests . METHODS: Samples from the introitus and the posterior vaginal fornix and vaginal lavage samples were collected from 91 women with a history suggestive of recurrent vulvovaginal candidosis (RVVC), and with a suspected new attack of the condition . The specimens were cultured on Sabouraud and CHROMagar . Speciation of yeast isolates was made on the chromogenic agar by API 32C kits and by an atomized system (Vitek) . RESULTS: Forty-six (51%) women were positive for Candida from one or more of the samples . The introital cultures were positive in 43 (47%) women, both on Sabouraud and chromogenic agar . From the posterior vaginal fomix, 42 (46%) women were positive on the Sabouraud and 43 (47%) on chromogenic agar cultures, while the vaginal lavage cultures yielded Candida on those two media in 40 (44%) and 41 (45%) cases, respectively . Candida albicans was the most frequent species recovered, from 40 (87%) cases, followed by C . krusei in 4 (9%), C . glabrata in 2 (4%), and C . parapsilosis in one case . There was only one woman who had a mixed yeast infection, by C . albicans and C . krusei . There was only one discrepancy in the speciation as demonstrated by mean of chromogenic agar and API 32C kit . CONCLUSIONS: Neither cultures of introital nor of vaginal lavage samples increases the detection rate of Candida in RVVC cases as compared to cultures of posterior vaginal fornix samples . Use of chromogenic agar is a convenient and reliable means to detect colonization by Candida and differentiate between C . albicans and non-albicans species.

J Investig Allergol Clin Immunol, 2002, 12(3), 182 - 91
Relevance of the selected cytokine release (TNF-alpha, IL-6, IFN-gamma, and IFN-alpha) to the exacerbation of bronchial asthma from airway mycotic infections . Predominant role of TFN-alpha?
Liebhart J, Cembrzynska-Nowak M, Bienkowska M, Liebhart E, Dobek R, Zaczynska E, Panaszek B, Obojski A, Malolepszy J.
Airway fungal infections are often associated in asthmatics with the exacerbation of asthma symptoms . However, the pathomechanism of this phenomenon has not been fully understood . The aim of our study was to assess whether antimycotic treatment can influence the capacity of bronchoalveolar (BAL) leukocytes to release proinflammatory cytokines, which could contribute to increase in asthma severity . Ten patients with bronchial asthma complicated by airway fungal infections (Candida albicans and/or Aspergillus fumigatus) were included in the study . Seven asthmatics were treated with systemic and inhaled corticosteroids, whereas the remaining three with inhaled ones only . All subjects underwent several courses of therapy with antibiotics due to respiratory infections . BAL leukocytes obtained from the patients were cultured in the absence or presence of lipopolysaccharide E.coli (LPS) or Newcastle disease virus (NDV) . The BAL procedure and measurement of the levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (II-6), interferon-gamma (IFN-gamma), and interferon-alpha (IFN-alpha) by specific bioassays were performed twice: before antimycotic treatment and after 3 weeks of therapy with 8 mg of nebulized fluoconazole and 400 mg of oral ketoconazole per day . The elimination of fungi from respiratory tract resulted in an apparent clinical improvement . This coincided with diminished production of TNF-alpha in response to LPS and the production of IFN-alpha in response to NDV, which were initially high and subsided significantly after antimycotic therapy (p = 0.035, and 0.011, respectively) . Such changes were not observed in the case of IFN-gamma and IL-6 . This may suggest that TNF-alpha as well as IFN-alpha are secreted by fungi-prestimulated leukocytes from the lower respiratory tract and may be involved in the processes of exacerbation of asthma complicated by fungal infections . Further analyses of relationships between changes in cytokine levels and clinical parameters indicated that IFN-alpha seems to be of particular interest in fungal stimulation of asthma.

APMIS, 2002 Sep, 110(9), 601 - 10
Candida glabrata, an emerging fungal pathogen, exhibits superior relative cell surface hydrophobicity and adhesion to denture acrylic surfaces compared with Candida albicans; Luo G et al.; Oral candidosis is a common opportunistic infection in debilitated individuals and Candida glabrata is the second most frequently isolated species from this condition, after Candida albicans . Candidal adherence to various biological or non-biological surfaces is considered a prerequisite for colonization, and pathogenesis of candidal infections, and their relative cell surface hydrophobicity (CSH) is likely to be a possible contributory force involved in this process . Whereas a large body of data on the latter features of C . albicans is available, there is surprisingly little information on C . glabrata . As a comprehensive database on the relative adhesion and CSH of Candida spp . is instructive and useful, we investigated in vitro the latter attributes of 34 oral isolates of C . glabrata and 15 isolates of C albicans . There were remarkable intraspecies differences in both the CSH and the adhesive ability of C . glabrata strains (p < 0.001) . Compared with C . albicans, C glabrata demonstrated a four-fold greater CSH value (30.63 +/- 11.20% vs 7.23+/-3.56%, p < 0.0001) and a two-fold greater tendency to adhere to denture acrylic surfaces (75.18 +/- 39.96 vs 30.36+/-9.21, p < 0.0001) . A significant positive correlation between CSH and adhesion was also noted for both C . glabrata (r=0.674, p < 0.0001) and C . albicans ( r = 0.636, p < 0.05) . When the effect of different incubation conditions on the relative CSH and adherence of C . glabrata was examined, CSH and the adherence to acrylic surfaces of four of six C . glabrata isolates were significantly affected by a reduction of the culture temperature (from 37 degrees C to 25 degrees C) . A positive relationship also emerged when the temperature-induced variations in the adherence values were correlated with their relative CSH . These data provide hitherto unavailable archival information on important pathogenic attributes of the two most common oral Candida species that may help explain their predominance in this milieu.

Mikrobiologiia, 2002 Nov-Dec, 71(6), 768 - 72
{The effect of sodium azide on the thermotolerance of the yeast Saccharomyces cerevisiae and Candida albicans}; Rikhvanov EG et al.; The addition of sodium azide (a mitochondrial inhibitor) at a concentration of 0.15 mM to glucosegrown Saccharomyces cerevisiae or Candida albicans cells before exposing them to heat shock increased cell survival . At higher concentrations of azide, its protective effect on glucose-grown cells decreased . Furthermore, azide, even at low concentrations, diminished the thermotolerance of galactose-grown yeast cells . It is suggested that azide exerts a protective effect on the thermotolerance of yeast cells when their energy requirements are met by the fermentation of glucose . However, when cells obtain energy through respiratory metabolism, the azide inhibition of mitochondria enhances damage inflicted on the cells by heat shock.

Genetics, 2002 Dec, 162(4), 1573 - 81
Candida albicans Mds3p, a conserved regulator of pH responses and virulence identified through insertional mutagenesis; Davis DA et al.; Candida albicans is a commensal fungus that causes diverse infections after antibiotic use or immune debilitation . Gene discovery has been limited because the organism is an asexual diploid . We have developed a strategy that yields random homozygous insertion mutants . The strategy has permitted identification of several prospective essential genes . Many of these genes are homologous to nonessential Saccharomyces cerevisiae genes, and some have no S . cerevisiae homolog . These findings may expand the range of antifungal drug targets . We have also identified new genes required for pH-dependent filamentation, a trait previously associated with virulence . One newly identified gene, MDS3, is required for expression in alkaline media of two filamentation-associated genes, HWP1 and ECE1, but is not required for expression of other pH-response genes . In S . cerevisiae, the two MDS3 homologs are required for growth in alkaline media, thus arguing that Mds3p function in adaptation to external pH changes is conserved . Epistasis tests show that Mds3p contributes to virulence and alkaline pH responses independently of the well-characterized Rim101p pH-response pathway.

Clin Infect Dis, 2003 Jan 15, 36(2), 221 - 4 Epub 2002 Dec 19.
Geographical differences in human oral yeast flora; Xu J et al.; The oral yeast flora of healthy humans from eastern North America and China were sampled and compared . Chinese persons harbored a greater number and diversity of yeast species in the mouth . Furthermore, Candida albicans, which is the predominant commensal and etiologic species of candidiasis in Europe and the Western Hemisphere, was relatively rare in China.

Med Mycol, 2002 Dec, 40(6), 557 - 63
Relative contributions of myeloperoxidase and NADPH-oxidase to the early host defense against pulmonary infections with Candida albicans and Aspergillus fumigatus; Aratani Y et al.; Generation of oxidative products by phagocytic cells is known to be an important host defense mechanism directed toward killing of invading microorganisms . The importance of two major oxidant-producing enzymes, myeloperoxidase (MPO) and NADPH-oxidase, in in vivo fungicidal action was directly compared in genetically engineered mice . Both MPO-deficient (MPO-/-) and NADPH-oxidase-deficient (X-linked chronic granulomatous disease {X-CGD}) mice showed increased susceptibility to pulmonary infections with Candida albicans and Aspergillus fumigatus compared with normal mice, and the X-CGD mice exhibited shorter survivals than MPO-/- mice . This increased mortality of X-CGD mice was associated with a 10- to 100-fold increased outgrowth of the fungi in their organs during the first 6 days . These results suggest that superoxide (O2-) produced by NADPH-oxidase is more important than hypochlorous acid (HOCl) produced by MPO, although both oxidative products obviously contribute to the host defense against pulmonary infection with those fungi . We also observed that MPO-/-/X-CGD double knockout mice showed comparable levels of susceptibility to the X-CGD mice against C . albicans and A . funigatus, indicating that MPO is unable to play a role in host defense in the absence of NADPH-oxidase . This strongly suggests that hydrogen peroxide, the precursor of HOCl, is solely derived from O2- produced by NADPH-oxidase.

Med Mycol, 2002 Dec, 40(6), 535 - 43
Evaluation of internal transcribed spacer region of ribosomal DNA sequence analysis for molecular characterization of Candida albicans and Candida dubliniensis isolates from HIV-infected patients; Millon L et al.; Molecular typing systems have been needed to study Candida colonization in HIV-infected patients, particularly for investigating virulence and fluconazole resistance . Three methods--electrophoretic karyotyping (EK), detection of restriction fragment length polymorphisms (RFLP) and randomly amplified polymorphic DNA analysis (RAPD)--have been most frequently used . In this study, comparative sequence analysis of the internal transcribed spacer (ITS) region of rDNA was evaluated for delineation of Candida isolates from 14 HIV-infected patients . EK, ITS sequence analysis, RFLP and RAPD resulted in 11, 10, 9 and 8 DNA genotypes, respectively, from 39 Candida albicans isolates . The 10 genotypes observed using ITS sequence analysis were defined by six variation sites in the sequence . Molecular typing of sequential oral isolates showed the persistence of the same genotype of C . albicans in nine patients, and genotype variation in one patient . EK and RAPD showed that another patient was co-infected by two distinct genotypes and ITS analysis identified one of the two genotypes as Candida dubliniensis . Comparative ITS sequence analysis is a quick and reproducible method that provides clear and objective results, and it also identifies C . dubliniensis . The discriminatory power of this new typing approach could be improved by concomitant analysis of other DNA polymorphic sequences.

Eur Arch Otorhinolaryngol, 2003 Jan, 260(1), 24 - 7 Epub 2002 Aug 01.
Concomitant otomycosis and dermatomycoses: a clinical and microbiological study; Ozcan M et al.; Otomycosis tends to recur despite long-term treatment . To our knowledge, there is no study in the English literature concerning the clinical importance of concomitant otomycosis and dermatomycoses . We investigated the presence of dermatomycoses in 52 patients with otomycosis in order to document the clinical and microbiological importance of their coexistence . Dermatomycoses on the feet and/or hands were identified in 19 patients (36.5%) . The most common pathogen for otomycosis was Aspergillus niger, while it was Candida albicans for the dermatomycoses . The same pathogenic fungi were isolated from the otomycosis and dermatomycoses in nine of the 19 patients (47.4%) . Aspergillus niger was the most common shared pathogen . The pathogens isolated in concomitant dermatomycoses were common pathogens for the fungal infection of the ear ( Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Candida albicans) . It was concluded that the autoinoculation of the ear canal by pathogenic fungi might be possible in the presence of the untreated dermatomycoses . Dermatomycoses must be investigated in patients with otomycosis and must be treated simultaneously in order to prevent the recurrence of both.

Yeast, 2003 Jan 30, 20(2), 149 - 56
Isolation and sequencing of the Candida albicans MSI3, a putative novel member of the HSP70 family; Cho T et al.; We have reported previously that the expression of CGR1 increased at an early stage of the yeast-mycelial transition (morphogenesis) in Candida albicans . We now show that Cgr1p interacts in a yeast two-hybrid system with the C . albicans Msi3p (CaMsi3p), a putative novel member of the heat shock protein 70 (HSP70) family . The DNA sequence of CaMSI3 encodes a predicted protein of 702 amino acids with a molecular mass of 78.6 kDa . The amino acid sequence of CaMsi3p is 63% identical to Msi3p/Sse1p of the HSP70 family of Saccharomyces cerevisiae . Further, CaMSI3 complemented the temperature-sensitive phenotype of the msi3(-) mutant of S . cerevisiae . Other heat shock proteins of C . albicans are required for morphogenesis and are highly antigenic . These observations suggest that CaMSI3 may well provide functions for this organism unrelated to a heat shock function . The DDBJ Accession No . for the sequence reported in this paper is AB061274 .

J Clin Microbiol, 2003 Jan, 41(1), 506 - 8
Uses and limitations of the XTT assay in studies of Candida growth and metabolism; Kuhn DM et al.; Colorimetric tetrazolium assays are used increasingly in studies of fungi, often in the absence of standardization or correlation with other methods . We examined species- and strain-related tetrazolium metabolism in Candida albicans and Candida parapsilosis by using XTT {2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-{(phenylamino) carbonyl}-2H-tetrazolium hydroxide} and WST-8 {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulphonyl)-2H-tetrazolium} and found marked variations . Also, significant signal was often missed in the absence of dimethyl sulfoxide extraction.

J Clin Microbiol, 2003 Jan, 41(1), 489 - 91
Evaluation of a method for identification of Candida dubliniensis bloodstream isolates; Sancak B et al.; To evaluate methods for differentiating Candida albicans and Candida dubliniensis, 772 putative C . albicans bloodstream isolates were tested for growth at 37 and 42 degrees C . Isolates showing no growth at 42 degrees C, abundant chlamydospore production, and the sugar assimilation pattern of the type strain were confirmed by DNA-based procedures to be C . dubliniensis.

J Clin Microbiol, 2003 Jan, 41(1), 475 - 8
Detecting Candida albicans in human milk; Morrill JF et al.; Procedures for diagnosis of mammary candidosis, including laboratory confirmation, are not well defined . Lactoferrin present in human milk can inhibit growth of Candida albicans, thereby limiting the ability to detect yeast infections . The inhibitory effect of various lactoferrin concentrations on the growth of C . albicans in whole human milk was studied . The addition of iron to the milk led to a two- to threefold increase in cell counts when milk contained 3.0 mg of lactoferrin/ml and markedly reduced the likelihood of false-negative culture results . This method may provide the necessary objective support needed for diagnosis of mammary candidosis.

J Clin Microbiol, 2003 Jan, 41(1), 414 - 20
Identification of Candida spp . by randomly amplified polymorphic DNA analysis and differentiation between Candida albicans and Candida dubliniensis by direct PCR methods; Bautista-Munoz C et al.; Because Candida species have innately highly variable antifungal susceptibilities, the availability of a fast and reliable species identification test is very important so that suitable and effective therapeutic measures may be taken . Using three oligonucleotide primers, we established a randomly amplified polymorphic DNA (RAPD) analysis method that enabled direct identification of the most common opportunistic pathogenic Candida species . RAPD analysis revealed a characteristic molecular fingerprint for each Candida species . Differences between the profiles for Candida albicans and C . dubliniensis were evident . RAPD analysis is a relatively easy, reproducible, and reliable technique that can be useful in providing genetic fingerprints for the identification of strains . In addition, a collection of different C . albicans strains was identified by a specific PCR based on multiple secreted aspartic proteinase (SAP) genes and the dipeptidyl aminopeptidase (DAP2) gene . Our findings demonstrate that PCR based upon the SAP and DAP2 sequences is a simple, rapid, clear, and direct technique for the identification and differentiation of C . albicans and C . dubliniensis.

Hand Clin, 2002 Nov, 18(4), 631 - 42, vi; discussion 643-6
Fungal infections of the perionychium; Yates YJ et al.; There are two types of fungi (yeasts and molds) both of which can cause superficial infections of the perionychium . Yeasts (such as Candida albicans) grow as single cells and reproduce by asexual budding . In contrast, molds grow in long filaments, called hyphae . There are approximately 100,000 species of fungi that have been characterized . Most of these are ubiquitous . Fortunately only about 200 are human pathogens, and only a handful are commonly found to be associated with human disease . This article discusses causes, symptoms, diagnosis, and treatment of the most common fungal infections of the perionychium, including superficial dermatophytosis, onychomycosis, and chronic paronychia.

Drugs Exp Clin Res, 2002, 28(4), 147 - 54
The SH-metabolite I of erdosteine, a mucolytic drug, enhances the inhibitory effect of salbutamol on the respiratory burst of neutrophils; Dal Sasso M et al.; Reactive oxygen species (ROS) are a common denominator of airway inflammation associated with chronic obstructive pulmonary disease (COPD) and asthma, as well as with less frequent lung diseases such as idiopathic pulmonary fibrosis (IPF), acute respiratory distress syndrome (ARDS) and cystic fibrosis (CF) . The most frequently administered drugs used to treat these diseases are bronchodilators, antioxidant/antiphlogistic agents and mucoactive drugs . The metabolization of the mucoactive drug erdosteine produces an active metabolite (Met I) with a reducing SH group . In addition to its mucolytic action, Met I also has useful antioxidant activity . The various activities of beta 2-agonists include their ability to reduce the respiratory burst of neutrophils and the subsequent release of ROS . beta 2-Agonists and mucoactive drugs may be administered to the same patients during the treatment of lung diseases . The aim of this study was to investigate the ability of Met I to potentiate the activity of salbutamol in inhibiting the in vitro respiratory burst of neutrophils by means of chemiluminescence . The combination of Met I 5 and 10 micrograms/ml with salbutamol 10(-5), 10(-6) and 10(-7) M led to a significant reduction in respiratory bursts when the neutrophils were stimulated with the soluble stimulant N-formyl-methionylleucyl-phenylalanine (fMLP) . The combinations of the two drugs that reduced the respiratory bursts when a particulate stimulus (Candida albicans) was used were those containing 10(-5) M of salbutamol . The reasons for this different behavior remain unclear and raise questions about the specific roles, sites and mechanisms of action of the different types of stimulation undergone by the respiratory airways.

Drugs Exp Clin Res, 2002, 28(4), 133 - 45
Inhibitory effects of zafirlukast on respiratory bursts of human neutrophils; Braga PC et al.; The effects of zafirlukast, a cysteinyl-leukotriene receptor antagonist, on the generation of the reactive oxygen species (ROS) released during respiratory bursts of human polymorphonuclear neutrophils (PMNs) is still unknown . The aim of this study was to investigate the ability of zafirlukast to interfere with the respiratory burst of PMNs . Respiratory burst responses of PMNs were investigated by luminol-amplified chemiluminescence (LACL) using particulate (Candida albicans and zymosan) and soluble stimulants {N-formyl-methionylleucyl-phenylalanine (fMLP) and phorbol 12 myristate 13 acetate (PMA)} . When incubated with PMNs for 10 min at concentrations ranging from 5 x 10(-9) M to 5 x 10(-6) M, zafirlukast did not significantly affect the respiratory bursts of PMNs induced by either the particulate or soluble stimuli . However, after incubation for 60 min, it did reduce the respiratory bursts of PMNs in a concentration-related fashion when the PMNs were stimulated with fMLP, and at a concentration of 5 x 10(-6) M when the stimulus was PMA . No significant effects were seen when the PMNs were challenged with particulate stimuli . Zafirlukast is able to interfere with the activation of the PMNs respiratory burst induced by soluble stimulants . The different behavior determined by different times of contact and different stimuli opens the way to interpretations concerning the antioxidant effect of zafirlukast.

Blood, 2003 Apr 15, 101(8), 3240 - 8 Epub 2003 Jan 02.
Human neutrophils utilize a Rac/Cdc42-dependent MAPK pathway to direct intracellular granule mobilization toward ingested microbial pathogens; Zhong B et al.; Elevated levels of mitogen-activated protein kinase/extracellular regulatory kinase (MAPK/ERK) activity are frequently found in some cancer cells . In efforts to reduce tumor growth, attempts have been made to develop cancer therapeutic agents targeting the MAPK . Here, by use of biologic, biochemical, and gene manipulation methods in human polymorphonuclear neutrophils (PMNs), we have identified a key pathway important in normal cell function involving MAPK/ERK in PMNs for growth inhibition of Candida albicans . Contact with C albicans triggered MAPK/ERK activation in PMNs within 5 minutes, and blocking of MAPK/ERK activation, either by the pharmacologic reagent PD098059 or by dominant-negative MAPK kinase (MEK) expression via vaccinia viral delivery, suppressed antimicrobial activity . Rac and Cdc42, but not Ras or Rho, were responsible for this MAPK/ERK activation . Expression of dominant-negative Rac (N17Rac) or Cdc42 (N17Cdc42) eliminated not only C albicans- mediated ERK phosphorylation but also phagocytosis and granule migration toward the ingested microbes, whereas dominant-negative Ras (N17Ras) and Rho (N19Rho) did not . PAK1 (p21-activated kinase 1) activation is induced by C albicans, suggesting that PAK1 may also be involved in the Rac1 activation of MAPK/ERK . We conclude from these data that Rac/Cdc42-dependent activation of MAPK/ERK is a critical event in the immediate phagocytic response of PMNs to microbial challenge . Therefore, use of MAPK pharmacologic inhibitors for the treatment of cancer may result in the interruption of normal neutrophil function . A balance between therapeutic outcome and undesirable side effects must be attained to achieve successful and safe anticancer therapy.

Arch Pharm Res, 2002 Dec, 25(6), 807 - 16
Flexible docking of an acetoxyethoxymethyl derivative of thiosemicarbazone into three different species of dihydrofolate reductase; Choi IH et al.; Dihydrofolate reductases (DHFR) of human, Candida albicans and E coli were docked with their original ligands of X-ray crystal complex using QXP (Quick eXPlore), a docking program . Conditions to reproduce the crystal structures within the root mean square deviation (rmsd) of 2.00 A were established . Applying these conditions, binding modes and species-specificities of a novel antibacterial compound, N4-(2-acetoxyethoxymethyl)-2-acetylpyridine thiosemicarbazone (AATSC), were studied . As the results, the docking program reproduced the crystal structures with average rmsd of six ligands as 0.91 A ranging from 0.49 to 1.45 A . The interactions including the numbers of hydrogen bonds and hydrophobic interactions were the same as the crystal structures and superposition of the crystal and docked structures almost coincided with each other . For AATSC, the results demonstrated that it could bind to either the substrate or coenzyme sites of DHFR in all three species with different degrees of affinity . It confirms the experimentally determined kinetic behavior of uncompetitive inhibition against either the inhibitor or the coenzyme . The docked AATSC overlapped well with the original ligands and major interactions were consistent with the ones in the crystal complexes . The information generated from this work should be useful for future development of antibacterial and antifungal agents.

Immunopharmacol Immunotoxicol, 2002 Nov, 24(4), 679 - 91
Fish immunology . II . Morphologycal and cytochemical characterization and phagocytic activities of head kidney macrophages from rainbow trout (Salmo gairdneri Richardson); Passantino L et al.; In Salmo gairdneri Richardson trouts, a comparison was made between macrophages (MO) derived from head kidney and peripheral blood monocytes . Morphologically and cytochemically no differences were observed between these two types of mononucleated cells . On the other hand, in parallel studies the ability of trout erythrocytes to engulf Candida albicans (CA) was evaluated and compared to the MO phagocytosis . In erythrocytes, engulfment is preceded by binding to CA and cell membrane invagination, while this was not the case for MO . Finally, MO stimulated with lipopolysaccharides (LPS) did not modify their phagocytic capacities, thus suggesting a lack of LPS receptors or a tolerance to LPS.

Immunopharmacol Immunotoxicol, 2002 Nov, 24(4), 665 - 78
Fish immunology . I . Binding and engulfment of Candida albicans by erythrocytes of rainbow trout (Salmo gairdneri Richardson); Passantino L et al.; The role of fish erythrocytes (FE) as phagocytic cells has poorly been investigated, until now . Here, we have focussed our attention on the interplay between rainbow trout (Salmo gairdneri Richardson) erythrocytes and Candida albicans (CA) . At the same time, the intervention of autologous head kidney macrophages (MO) in the CA processing by FE has been studied . Data show that CA particles bind to FE, which, in turn, are able to engulf but not kill them . In the presence of MO, a decrease of FE with bound CA occurs and, in some microscopic images, FE form rosettes with MO . Phagocytosis of CA is higher in rosetting MO than in non-rosetting ones . According to our findings, it appears that FE represent a reservoir of engulfed CA and rosetting is an efficacious phenomenon of presentation of pathogens to MO, where an effective clearance of them can take place.

Int J Antimicrob Agents, 2003 Jan, 21(1), 20 - 6
Antimicrobial properties of highly fluorinated bis-ammonium salts; Massi L et al.; Four series of new highly fluorinated bisammoniums (Quaterfluo Tx, Quaterfluo Bx, Quaterfluo Cx, Quaterfluo Dx) were tested to evaluate their anti-bacterial and anti-fungal properties . Four microbial strains were used to perform the study: Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans and Aspergillus niger . The antimicrobial efficacy was measured by bacterial and fungal growth inhibition expressed as minimal inhibitory concentration (MIC) values . The Quaterfluo Tx series had very good antimicrobial activity, particularly against Gram-negative bacteria . A speed of kill test carried out with Quaterfluo T3 showed bactericidal activity . The introduction of perfluorinated chains into the quaternary ammonium surfactant structure leads to particularly active antimicrobial agents which not only have bacteriostatic properties but which could be also powerful bactericides.

Invest Ophthalmol Vis Sci, 2003 Jan, 44(1), 210 - 6
Experimental keratomycosis in a mouse model; Wu TG et al.; PURPOSE: To establish a murine model of corneal candidiasis that permits molecular evaluation of fungal adherence and invasion . METHODS: Corneas of immunocompetent, methylprednisolone-treated, and cyclophosphamide-treated adult NIH Swiss and BALB/c mice were topically mock inoculated or inoculated with 10-fold increasing amounts between 100 and 100 million colony-forming units (CFU) of Candida albicans after unilateral corneal scarification . Mock-inoculated eyes served as the control . Eyes were scored daily on a 12-point scale to categorize corneal inflammation and were enucleated for quantitative fungal cultures, analysis by polymerase chain reaction (PCR), and histopathologic examination . RESULTS: At least 100 CFU of C . albicans initiated measurable corneal infection, but 1 million or more colony-forming units were needed to induce consistent keratitis . Treatment with methylprednisolone increased disease severity in infected BALB/c mice and fungal persistence in both BALB/c and NIH Swiss mice . Treatment with cyclophosphamide increased disease severity and fungal persistence in both strains of mice . Infectious organisms were recovered by quantitative culture, and candidal DNA was detectable by PCR . C . albicans, inflammatory cells, and stromal necrosis were histologically evident within ocular tissue . CONCLUSIONS: Although mice are innately resistant to Candida infection after corneal inoculation, moderate to severe keratomycosis can be established in immunocompromised mice by the route of corneal scarification . Although differences between mouse strains and among immunosuppressive regimens remain to be explored, this murine model provides the basis for understanding the pathogenesis of fungal infections of the cornea.

Folia Microbiol (Praha), 2002, 47(5), 488 - 92
In vitro antifungal activity of 3-phenyl-2H-benzoxazine-2,4(3H)-diones; Waisser K et al.; A series of 81 3-phenyl-2H-benzoxazine-2,4(3H)-diones with substitution at C(6) on the benzoxazine ring and on the phenyl moiety was synthesized; the compounds were evaluated for antifungal activity against five strains of potentially pathogenic fungi (Absidia corymbifera, Aspergillus fumigatus, Candida albicans, Microsporum gypseum and Trichophyton mentagrophytes) . Structure-activity relationships against T . mentagrophytes and M . gypseum were determined using the Free-Wilson method, which was further combined with the approach of Hansch . In vitro antifungal activity becomes higher with increasing electron-accepting ability of the substituents on the phenyl ring, and with increasing lipophilicity.

J Nat Prod, 2002 Dec, 65(12), 1815 - 9
Five new steroidal saponins from Solanum chrysotrichum leaves and their antimycotic activity; Zamilpa A et al.; Using bioactivity-directed isolation procedures, five new spirostan saponins and two sterol glycosides have been isolated from Solanum chrysotrichum leaves . The structure of these compounds was established based upon spectroscopic measurements, especially 1D and 2D NMR data of their peracetate derivatives . These compounds showed antimycotic activity . The most active compound is 6alpha-O-beta-d-xylopyranosyl-(1-->3)-beta-d-quinovopyranosyl-(25R)-5alpha-spirostan-3beta,23alpha-ol (2) (MIC =12.5, 12.5, 100, and 200 microg/mL against Trichophyton mentagrophytes, T . rubrum, Aspergillis niger, and Candida albicans, respectively).

Antimicrob Agents Chemother, 2003 Jan, 47(1), 262 - 7
Synergistic activity of the N-terminal peptide of human lactoferrin and fluconazole against Candida species; Lupetti A et al.; In light of the need for new antifungal regimens, we report that at noncandidacidal concentrations, the lactoferrin-derived peptide hLF(1-11), which is highly active against fluconazole-resistant Candida albicans, acts synergistically with fluconazole against this yeast and a fluconazole-sensitive C . albicans strain as well as C . glabrata, C . krusei, C . parapsilosis, and C . tropicalis . When these yeasts were exposed to hLF(1-11) for 5 min and then incubated with fluconazole, they were killed effectively, while no candidacidal activity was observed when they were incubated first with fluconazole and then exposed to the peptide, indicating that the candidacidal activity is initiated by the peptide while fluconazole is only required during the effector phase . Investigations of the effect of azide, which inhibits mitochondrial respiration, on the activity of combinations of hLF(1-11) and fluconazole against fluconazole-resistant C . albicans revealed that it inhibits this activity, even when added during the effector phase only . As expected, azide inhibited the accumulation of rhodamine 123 in mitochondria and the production and release of ATP by C . albicans that occurred upon exposure to the combination of hLF(1-11) and fluconazole . Accordingly, oxidized ATP (oATP), an antagonist of ATP receptors, completely blocked the candidacidal activity of the hLF(1-11)-fluconazole combination, whereas oATP did not block the activity when its presence was restricted to the effector phase . The candidacidal activity of combinations of hLF(1-11) and fluconazole, which is initiated by the peptide through the involvement of energized mitochondria, renders fluconazole-resistant C . albicans sensitive to this azole.

Antimicrob Agents Chemother, 2003 Jan, 47(1), 34 - 8
Fluconazole susceptibility of vaginal isolates obtained from women with complicated Candida vaginitis: clinical implications; Sobel JD et al.; Despite considerable evidence of azole resistance in oral candidiasis due to Candida species, little is known about the azole susceptibilities of the genital tract isolates responsible for vaginitis . The fluconazole susceptibilities of vaginal isolates obtained during a multicenter study of 556 women with complicated Candida vaginitis were determined by evaluating two fluconazole treatment regimens . Of 393 baseline isolates of Candida albicans, 377 (96%) were highly susceptible to fluconazole (MICs, <8 microg/ml) and 14 (3.6%) were resistant (MICs, >or=64 microg/ml) . Following fluconazole therapy, one case of in vitro resistance developed during 6 weeks of monitoring . In accordance with the NCCLS definition, in vitro fluconazole resistance correlated poorly with the clinical response, although a trend of a higher mycological failure rate was found (41 versus 19.6% on day 14) . By using an alternative breakpoint of 1 micro g/ml, based upon the concentrations of fluconazole achievable in vaginal tissue, no significant differences in the clinical and mycological responses were observed when isolates (n = 250) for which MICs were <or=1 microg/ml were compared with isolates (n = 30) for which MICs were >1 microg/ml, although a trend toward an improved clinical outcome was noted on day 14 (odds ratio, >2.7; 95% confidence interval, 0.91, 8.30) . Although clinical failure was uncommon, symptomatic recurrence or mycological relapse almost invariably occurred with highly sensitive strains (MICs, <1.0 microg/ml) . In vitro fluconazole resistance developed in 2 of 18 initially susceptible C . glabrata isolates following fluconazole exposure . Susceptibility testing for women with complicated Candida vaginitis appears to be unjustified.

J Ethnopharmacol, 2003 Jan, 84(1), 1 - 4
In vitro antimicrobial activity of ethanol and water extracts of Cassia alata; Somchit MN et al.; Crude ethanol and water extract of leaves and barks from Cassia alata were tested in vitro against fungi, (Aspergillus fumigatus and Microsporum canis), yeast (Candida albicans) and bacteria (Staphylococcus aereus and Escherichia coli) . C . albicans showed concentration-dependent susceptibility towards both the ethanol and water extracts from the barks, but resistant towards the extracts of leaves . The degree of susceptibility varied, the water extract from barks showed bigger inhibition zone than the ethanol extracts (12-16 and 10-14 mm, diameter respectively) . The growth of Aspergillus fumigatus and Microsporum canis were not affected by all types of the plant extracts . Results were comparable to standard antifungal drug Tioconazole (18 mm diameter) at equivalent concentration . The anti-bacterial activity of C . alata extracts on S . aureus was detected with only the leaves extracts using water and ethanol . The water extract exhibited higher antibacterial activity than the ethanol extract from leaves (inhibition zones of 11-14 and 9-11 mm, respectively) . E . coli showed resistance to all types of extracts . Based on the current findings, it can be concluded that this plant has antimicrobial activity, which is as potent as standard antimicrobial drugs against certain microorganisms .

Infect Immun, 2003 Jan, 71(1), 437 - 45
Normal host defense during systemic candidiasis in mannose receptor-deficient mice; Lee SJ et al.; Pathogen pattern recognition receptors (PRRs) recognize common structural and molecular motifs present on microbial surfaces and contribute to induction of innate immune responses . The mannose receptor (MR), a carbohydrate-binding receptor expressed on subsets of macrophages, is considered one such PRR . In vitro experiments have implicated the MR in phagocytosis of mannose-bearing microbes, including Candida albicans, and enhancement of antifungal response by macrophages . However, the significance of the MR's contribution to immune response during systemic C . albicans infection has never been directly demonstrated . Using MR-deficient mice in an in vivo infection experiment, we examined the role of the MR in immune response during disseminated candidiasis . MR(-/-) and wild-type control mice were challenged intraperitoneally with C . albicans, and the survival rates, tissue fungal burden, inflammatory cell recruitment, and specific antibody production after infection were evaluated . We found no significant difference in survival between the two mouse strains . MR(-/-) mice had higher average fungal burdens in some of the organs on days 7 and 21 but exhibited competence in inflammatory cell recruitment and antibody production . We also observed in vitro that MR(-/-) peritoneal cavity macrophages were equally capable of C . albicans uptake and that phagocytosis could be blocked with beta-glucan . We conclude that the MR is not required for the normal host defense during disseminated candidiasis or for the phagocytosis of C . albicans and that a beta-glucan receptor may be required for C . albicans phagocytosis.

Mol Microbiol, 2003 Jan, 47(1), 89 - 102
EFG1 is a major regulator of cell wall dynamics in Candida albicans as revealed by DNA microarrays; Sohn K et al.; Cell wall dynamics in Candida albicans, the most common fungal pathogen in man, underlie regulatory processes during the yeast-to-hyphae transition . To analyse this regulation at the transcriptional level, we have established a DNA microarray representing genes implicated in cell wall biogenesis . Using these microarrays, we were able to identify YWP1 and HWP2 that are specifically transcribed in the yeast or hyphal growth form respectively . Cluster analysis revealed at least two major clusters of genes: cluster I comprised genes that were upregulated under at least one hyphae-inducing condition . Three as yet not further characterized genes were attributed to cluster II . These genes were transcribed in the yeast form of C . albicans and were downregulated in an EFG1-dependent manner under specific hyphae-inducing conditions . We show further that, in contrast to CPH1, EFG1 plays a major role in the transcriptional regulation of cell wall proteins under the conditions investigated . EFG1 was essential for the transcription of both hyphae-specific genes such as HWP1 and HWP2 as well as the yeast form-specific gene YWP1 . Moreover, we found that, under various conditions, EFG1 also can act as a strong repressor for the transcription of RBE1, another not yet characterized cell wall protein . Overall, our data show that EFG1 plays a major role in the induction and repression of cell wall genes, not only in the hyphal form but also in the yeast form of C . albicans.

Fitoterapia, 2002 Dec, 73(7-8), 651 - 62
Light-mediated activities of some Thai medicinal plant teas; Cheeptham N et al.; Forty-one types of commercially available Thai medicinal teas were tested for their light-mediated activities against Bacillus subtilis, Staphylococcus aureus K147 methicillin-sensitive (Ms), Escherichia coli DC10, E . coli (wild), Pseudomonas aeruginosa 187 (wild), Candida albicans and Aspergillus fumigatus . The results represent a first report of the light-mediated antimicrobial activities of Thai medicinal teas and suggest that phytochemical investigations may be warranted .

Can J Microbiol, 2002 Oct, 48(10), 886 - 94
Effect of a combination therapy between IL-12 and soluble IL-4 receptor (sIL-4R) on Candida albicans and herpes simplex virus type I infections in thermally injured mice; Kobayashi M et al.; The effectiveness of a combination using IL-12 and soluble IL-4 receptor (sIL-4R) to treat severe infections of herpes simplex virus type 1 (HSV-1) and Candida albicans in thermally injured mice was investigated . Although sIL-4R decreased burn-associated type 2 T-cell responses, the effect of sIL-4R was minimal on the morbidity and mortality of thermally injured mice exposed to 250 times LD50 of HSV-1 or 10 times LD50 of C . albicans . Compared with 100% mortality in control mice, mortality for HSV-1 and C . albicans was 40 and 20%, respectively, in thermally injured mice that received IL-12 and sIL-4R in combination . After stimulation with anti-CD3 monoclonal antibody, splenic T cells from thermally injured mice exposed to large amounts of HSV-1 or C . albicans did not produce gamma interferon (IFN-gamma) into their culture fluids . However, IFN-gamma was produced by splenic T cells from thermally injured and infected mice treated with IL-12 and sIL-4R in combination . These results suggest that therapeutic treatment with a combination of IL-12 and sIL-4R may be effective by inducing type 1 T-cell responses in thermally injured mice exposed to large amounts of HSV-1 or C . albicans.

Oral Microbiol Immunol, 2002 Dec, 17(6), 375 - 8
Cytokines in the oral mucosa of mice infected with Candida albicans; Farah CS et al.; Oropharyngeal candidiasis is associated with defects in cell-mediated immunity, and is commonly seen in immunocompromised patients . We have previously shown that T-cell-deficient BALB/c nude (nu/nu) mice are extremely susceptible to oropharyngeal candidiasis, and that recovery from a chronic infection is dependent on CD4 T lymphocytes . In this study we describe the local tissue cytokine profile in lymphocyte-reconstituted immunodeficient mice and their euthymic counterparts . Mice were infected orally with 108 cells of the yeast Candida albicans, and oral tissues sampled on days 0, 4, 8, and 14 . Nude mice were reconstituted with 3 x 107 naive lymphocytes following oral inoculation . Interleukin (IL)-6, interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha were identified in the oral tissues of infected euthymic mice recovering from oral infection, as well as naive controls . TNF-alpha was identified in nude oral tissue on days 4 and 8, but only after lymphocyte reconstitution . No IL-2, IL-4 or IL-10 was detected in either euthymic or athymic mice at any time-point throughout the experiment . This study confirms the functional activity of T lymphocytes in reconstituted nude mice, and suggests that TNF-alpha may be an important mediator in the recovery from oropharyngeal candidiasis.

Clin Oral Investig, 2002 Dec, 6(4), 227 - 35 Epub 2002 Oct 15.
Quantitative determination of salivary components in the pellicle on PMMA denture base material; Gocke R et al.; The formation of a salivary pellicle is a protective mechanism of the body for all surfaces in the oral cavity . The nature of the substrate may influence the composition of the pellicle . The aim of this study was to investigate the quantitative composition and individual variation of the salivary pellicle formed on denture base material (PMMA) . Cylindrical specimens of PMMA were carried in the mouth and then desorbed with a 0.5-M sodium chloride solution . The solution was analysed for total protein, alpha-amylase, total proteases, protease inhibitors, secretory immunoglobulin A, immunoglobulin G, peroxidases, thiocyanate, lysozyme, and calcium content . All investigated salivary components could be found unequivocally in the desorption solution, indicating that a salivary pellicle had formed on the surface of the PMMA . Large coefficients of variation indicate large individual variations in the adsorbed amounts . The data also point to large intraindividual variations for the bound salivary components . Only the protease inhibitors revealed a strong positive correlation of the bound activity to the salivary activity . It is hypothesised that differences in the bound amounts of antimicrobial components might influence the microbial colonisation of denture bases and that protease inhibitors could be meaningful for the spread of the yeast Candida albicans from denture base material to the oral mucosa and thus might be an explanation for different susceptibility to denture base stomatitis.

Eukaryot Cell, 2002 Dec, 1(6), 1010 - 20
Deletion of individual mRNA capping genes is unexpectedly not lethal to Candida albicans and results in modified mRNA cap structures; Dunyak DS et al.; Eukaryotic mRNAs are modified at the 5' end with a cap structure . In fungal cells, the formation of the mRNA cap structure is catalyzed by three enzymes: triphosphatase, guanylyltransferase, and methyltransferase . Fungal capping enzymes have been proposed to be good antifungal targets because they differ significantly from their human counterparts and the genes encoding these enzymes are essential in Saccharomyces cerevisiae . In the present study, Candida albicans null mutants were constructed for both the mRNA triphosphatase-encoding gene (CET1) and the mRNA methyltransferase encoding gene (CCM1), proving that these genes are not essential in C . albicans . Heterozygous deletions were generated, but no null mutants were isolated for the guanylyltransferase-encoding gene (CGT1), indicating that this gene probably is essential in C . albicans . Whereas these results indicate that Cet1p and Ccm1p are not ideal molecular targets for development of anticandidal drugs, they do raise questions about the capping of mRNA and translation initiation in this fungus . Southern blot analysis of genomic DNA indicates that there are not redundant genes for CET1 and CCM1 and analysis of mRNA cap structures indicate there are not alternative pathways compensating for the function of CET1 or CCM1 in the null mutants . Instead, it appears that C . albicans can survive with modified mRNA cap structures.

Eukaryot Cell, 2002 Dec, 1(6), 967 - 77
Analysis of telomerase in Candida albicans: potential role in telomere end protection; Singh SM et al.; Telomerase is a ribonucleoprotein reverse transcriptase responsible for the maintenance of one strand of telomere terminal repeats . Analysis of the telomerase complex in the budding yeast Saccharomyces cerevisiae has revealed the presence of one catalytic protein subunit (Est2p/TERT) and at least two noncatalytic components (Est1p and Est3p) . The genome of the pathogenic yeast Candida albicans contains putative orthologues of all three telomerase components . Disruption of each homologue resulted in significant but distinct telomere dysfunction in Candida: Similar to S . cerevisiae, the Candida EST3 disruption strain exhibits progressive telomere loss over many generations, at a rate that is consistent with incomplete replication . In contrast, telomeres in both the Candida TERT and EST1 disruption strains can contract rapidly, followed by partial or nearly complete recovery, suggesting a defect in telomere "capping." We propose that these two telomerase subunits may participate in the protection of chromosomal ends in Candida: Analysis of telomerase-mediated primer extension in vitro indicates that only the TERT protein is absolutely essential for enzyme activity . Our results support the conservation of telomerase protein components beyond the catalytic subunit but reveal species-specific phenotypic alterations in response to loss of individual telomerase component . We also identify potential homologues of Est1p in phylogenetically diverse organisms . The Est1p sequence family possesses a conserved N-terminal domain predicted to be structurally related to tetratricopeptide repeat-containing proteins.

Eukaryot Cell, 2002 Dec, 1(6), 865 - 74
Gpa2, a G-protein alpha subunit required for hyphal development in Candida albicans; Sanchez-Martinez C et al.; Candida albicans is able to respond to environmental changes by inducing a distinct morphological program, which is related to the ability to infect mammalian hosts . Although some of the signal transduction pathways involved in this response are known, it is not clear how the environmental signals are sensed and transmitted to these transduction cascades . In this work, we have studied the function of GPA2, a new gene from C . albicans, which encodes a G-protein alpha-subunit homologue . We demonstrate that Gpa2 plays an important role in the yeast-hypha dimorphic transition in the response of C . albicans to some environmental inducers . Deletion of both alleles of the GPA2 gene causes in vitro defects in morphological transitions in Spider medium and SLAD medium and in embedded conditions but not in medium containing serum . These defects cannot be reversed by exogenous addition of cyclic AMP . However, overexpression of HST7, which encodes a component of the filament-inducing mitogen-activated protein kinase (MAPK) cascade, bypasses the Gpa2 requirement . We have obtained different gain-of-function and loss-of-function mutant alleles of the GPA2 gene, which we have introduced in several C . albicans genetic backgrounds . Our results indicate that, in response to environmental cues, Gpa2 is required for the regulation of a MAPK signaling pathway.

Eukaryot Cell, 2002 Dec, 1(6), 856 - 64
Hyphal tip-associated localization of Cdc42 is F-actin dependent in Candida albicans; Hazan I et al.; The rho-type GTPase Cdc42 is important for the establishment and maintenance of eukaryotic cell polarity . To examine whether Cdc42 is regulated during the yeast-to-hypha transition in Candida albicans, we constructed a green fluorescence protein (GFP)-Cdc42 fusion under the ACT1 promoter and observed its localization in live C . albicans cells . As in Saccharomyces cerevisiae, GFP-Cdc42 was observed around the entire periphery of the cell . In yeast-form cells of C . albicans, it clustered to the tips and sides of small buds as well as to the mother-daughter neck region of large-budded cells . Upon hyphal induction, GFP-Cdc42 clustered to the site of hyphal evagination and remained at the tips of the hyphae . This temporal and spatial localization of Cdc42 suggests that its activity is regulated during the yeast-to-hypha transition . In addition to the accumulation at the hyphal tip, GFP-Cdc42 was also seen as a band within the hyphal tube in cells that had undergone nuclear separation . With the F-actin-assembly inhibitor latrunculin A, we found that GFP-Cdc42 accumulation at the bud site in yeast-form cells is F-actin independent, whereas GFP-Cdc42 accumulation at the hyphal tip requires F-actin . Furthermore, disruption of the F-actin cytoskeleton impaired the transcriptional induction of hypha-specific genes . Therefore, hypha formation resembles mating in Saccharomyces cerevisiae in that both require F-actin for GFP-Cdc42 localization and efficient signaling.

Scand J Infect Dis, 2002, 34(10), 778 - 80
Successful treatment of Candida glabrata myocarditis with voriconazole; Einarsdottir HM et al.; A 30-y-old man with Crohn's disease developed fungemia with Candida albicans . Subsequently, during therapy with fluconazole, Candida glabrata was repeatedly isolated from his blood . Myocardial abscesses were detected in the papillary muscles and interventricular septum . The infection was cured with amphotericin B lipid complex and 5-flucytosine, followed by voriconazole for 18 months.

Mikrobiyol Bul, 2002 Jan, 36(1), 65 - 9
{Candida colonization on the surface of orthodontic brackets and the adhesion of these strains to buccal epithelial cells}; Gokdal I et al.; Many types and numbers of microorganisms may be colonized in the oral cavity . Candida albicans is the most frequently isolated fungal species which colonizes oral mucosal cells Denture prostheses play an important role by increasing the risk of colonization . In this study, Candida colonization and adhesion rates have been investigated among 60 orthodontic bracket users (mean age: 17.8 years), and 15 (25%) of them were found to be colonized by Candida species (13 C . albicans, 1 C . kefyr, 1 C . lusitaniae) . Fourteen of 15 subjects (93.3%) were between 16-18 years old . All the isolates were exposed with buccal epithelial cells in-vitro, and the adhesion rate was found to be 12% . The adhesion activity was detected only in C . albicans strains.

Mikrobiyol Bul, 2002 Jan, 36(1), 57 - 63
{Candida dubliniensis studies and isolation of Candida types in oropharyngeal specimens from oncologic patients}; Tekeli A et al.; Fungal opportunistic infections, and in particular those caused by the various Candida species, have gained considerable significance as a cause of morbidity and, often, mortality . Although Candida albicans remains to be the most frequently isolated fungal species as an opportunistic oral pathogen, other yeast species are often identified in immunocompromised patients . C . dubliniensis, the recently described species, has been recovered primarily from oropharyngeal candidasis in Human Immunodeficiency Virus (HIV)-infected patients . C . dubliniensis shares many phenotypic characteristics with, and is phylogenetically closely related to, C . albicans . The aim of the present study was to investigate the colonization rates of fungal species, and especially C . dubliniensis, in the oropharyngeal samples from cancer patients . The oropharyngeal swabs of 543 patients were collected during their visits to oncology clinic in 9 months period, and a total of 209 Candida species have been isolated . Of them, 147 isolates were found to be positive for germ tube and chlamydospore formation, and they were tested for the growth inability at 42 degrees C and 45 degrees C, colony morphology in Staib agar and the intracellular beta-glucosidase activity, in order to identify C . dubliniensis . The results of these tests and carbohydrate assimilation tests by API 20C AUX yeast identification system, yielded that all these 147 (70.3%) isolates were C . albicans . The other isolates were identified as follows; 16 C . parapsilosis (7.6%), 13 C . tropicalis (6.2%), 10 C . glabrata (4.7%), 5 C . guilliermondii (2.3%), 4 C . krusei (1.9%), 3 C . keyfr (1.4%), 3 C . famata (1.4%), 2 S . cerevisiae (0.9%), 2 C . pelliculosa (0.9%), 1 C . utiles (0.4%), 1 C . neoformans (0.4%) and 1 Hansenula polymorpha (0.4%), while no C . dubliniensis was isolated.

J Pathol, 2003 Jan, 199(1), 98 - 106
Differential expression of CCL19 by DC-Lamp+ mature dendritic cells in human lymph node versus chronically inflamed skin; Katou F et al.; De novo formation of lymphoid tissue is one of the characteristic features of chronic inflammation . The formation of T cell-mature dendritic cell (DC) clusters has been previously demonstrated in chronically inflamed skin infected with Candida albicans . A functional similarity was also found between chronic inflammation and the T-cell zone of lymph nodes (LNs), since a substantial fraction of phenotypically mature DCs in both tissues expressed CCL22 (macrophage-derived chemokine; MDC) and were closely surrounded by memory-type T cells expressing its receptor, CCR4 . To analyse the nature of T cell-mature DC interactions further in chronically inflamed skin and LNs, the present study focuses on another chemokine system, namely CCL19 (EBI1 ligand chemokine; ELC), CCL21 (secondary lymphoid tissue chemokine; SLC) and their shared receptor, CCR7 . RT-PCR analysis revealed expression of CCL19, CCL21, and CCR7 at high levels in LNs and at low levels in inflamed skin . Using immunohistochemistry, the majority of DC-Lamp(+) mature DCs in the T-cell area of LNs expressed CCL19 and were surrounded by CCR7(+) naive-type lymphocytes, while CCL21 was expressed in reticular stromal cells and vascular endothelial cells . Very few mature DCs in LNs were found to express CCR7 . In contrast, the majority of DC-Lamp(+) mature DCs in inflamed skin were totally negative for CCL19 and were surrounded by CCR7(-) memory-type T cells . Furthermore, CCL21 expression in the inflamed skin was detected in dermal lymphatic endothelial cells and rare CCR7(+) mature DCs were mostly seen within the lymphatic vessels . In normal skin, on the other hand, no cells immunoreactive for CCL19, CCL21, or CCR7 were found . The present study thus reveals a striking difference in the function of mature DCs between LNs and chronically inflamed skin .

Mycoses, 2002 Dec, 45(11-12), 461 - 4
Semi-quantitative adsorption for detection of antibodies to Candida albicans germ tubes by indirect immunofluorescence test; Bulajic N et al.; Adsorption of antibodies against antigens expressed on the Candida albicans blastoconidia cell walls was standardized for the detection of antibodies to germ tubes by an indirect immunofluorescence test (IIFT) . Sera from rabbits immunized with C . albicans bearing germ tubes, were diluted by two-fold serial dilutions to obtain specimens 1 : 640-1 : 20 positive by IIFT . Different volumes of specimens were adsorbed with different amounts of whole, heat-inactivated C . albicans blastoconidia . It was found that a 1 : 640 titre serum should be adsorbed at 30 microl by 50 mg cells, down to a 1 : 20 titre serum at 110 microl by 12.5 mg cells . Accurate detection of anti-germ tube antibodies by IIFT depends on the semi-quantitative relation between the positive titre of the specimen and the amount of blastoconidia used in adsorption.

Clin Infect Dis, 2002 Dec 15, 35(12), 1477 - 83 Epub 2002 Dec 04.
Molecular diversity and routes of colonization of Candida albicans in a surgical intensive care unit, as studied using microsatellite markers; Stephan F et al.; To evaluate the colonization of Candida species and the importance of cross-contamination with Candida albicans, we prospectively screened clinical specimens obtained from surgical patients in the intensive care unit (ICU) who had a high risk of yeast colonization . Genotyping of C . albicans was performed using microsatellite markers . Thirty-six of 94 patients acquired nosocomial yeast colonization and/or infection . A total of 1126 specimens were cultured, 167 (15%) of which yielded yeasts . All 122 isolates of C . albicans recovered from the 30 C . albicans-positive patients were genotyped . Twenty-four different genotypes were identified . No genotype was systematically associated with a specific room or time . Isolates recovered from different body sites of patients at different times had identical genotypes . Acquisition of C . albicans in the surgical ICU seems to be mainly endogenous . Microsatellite markers should also be developed for typing non-albicans Candida species to learn whether their epidemiology differs from that of C . albicans.

Endoscopy, 2002 Dec, 34(12), 941 - 5
Eight years' experience of gastrostomy tube management; Koulentaki M et al.; BACKGROUND AND STUDY AIMS: Increasingly, patients fed by gastrostomy tube are surviving the lifespan of the device . Data are scarce concerning the factors affecting the longevity and failure of gastrostomy tubes or the criteria for selection of replacement devices which leads to cost-effective patient management . The aims of the study were: to set criteria for selection of replacement gastrostomy tubes; to determine the causes of gastrostomy tube failure, and the factors affecting device longevity; and to examine the effect of initiating an educational programme for caregivers on resource utilization in long-term enteral nutrition patients . MATERIALS AND METHODS: We analyzed the clinical gastrostomy tube database compiled prospectively over 8 years by the nutrition team at Ninewells Hospital, Dundee . RESULTS: For 363 gastrostomy tubes inserted in 304 patients (160 women; median age 71), the median duration of gastrostomy tube use was 138 days . The total follow-up was 294 patient-years . Death occurred before the first gastrostomy tube replacement in 48 % of patients, but 20 % resumed oral nutrition . Tube failure mechanisms were: dislodgment, 28 %; perishing of tube material, 25 %; tube-related Candida albicans infection, 16 %; leakage, 7 %; and unspecified, 7 % . Of the balloon tubes and gastrostomy buttons, 8 % needed early replacement due to dislodgment and/or leakage . The cost per day for replacement percutaneous endoscopic gastrostomy (PEG) was Euro 2.12, for balloon tubes it was Euro 0.62, and for gastrostomy buttons Euro 1.80 . Despite an increasing PEG insertion rate throughout the study period, yearly referrals for PEG-related problems dropped by 30 % between 1997 to 1999, coinciding with the initiation of an educational programme for caregivers . CONCLUSION: Tube longevity is mainly limited by the patient's diagnosis and prognosis . The choice of replacement device should be based on clinical factors . The use of more durable materials in the manufacture of gastrostomy tubes may prolong tube life and reduce cost . Education of patients and caregivers by a multidisciplinary nutrition support team promotes independence and limits demand on the service.

Mol Genet Genomics, 2002 Dec, 268(4), 468 - 76 Epub 2002 Nov 16.
Sec20p-interacting proteins (Tip20p, Ufe1p) in the retrograde secretory pathway of the fungal pathogen Candida albicans; Weber Y et al.; Sec20p is an essential Type-II membrane protein of the human fungal pathogen Candida albicans, which is thought to be involved in mediating retrograde vesicle traffic from the Golgi to the endoplasmic reticulum (ER) . Using an epitope-tagged Sec20p we obtained evidence for its localization in ER membranes, which is consistent with its proposed role in an ER-tSNARE complex . Two genes encoding potential interaction partners for Sec20p, Tip20p and Ufe1p, were identified in genomic sequences of C . albicans; these show 18% and 27% identity, respectively, to homologues in Saccharomyces cerevisiae . An interaction between the cytoplasmic domain of Sec20p and Tip20p was demonstrated by two-hybrid analysis; in addition, Tip20p was found to form homodimers . Interaction between Sec20p and Tip20p in vivo was verified by co-immunoprecipation experiments . CaUFE1, which encodes a potential ER-tSNARE, was able to complement a thermosensitive ufe1 mutation in S . cerevisiae, suggesting functional conservation between the two fungal proteins . Thus, although the sequences of some components of the ER-tSNARE complex have diverged considerably during evolution, it appears that they have retained similar functions in C . albicans and S . cerevisiae.

Mem Inst Oswaldo Cruz, 2002 Oct, 97(7), 985 - 9
First characterization of Candida albicans by random amplified polymorphic DNA method in Nicaragua and comparison of the diagnosis methods for vaginal candidiasis in Nicaraguan women; Darce Bello M et al.; A total of 106 women with vaginitis in Nicaragua were studied . The positive rate for the identification of Candida species was 41% (44 positive cultures out of 106 women with vaginitis) . The sensitivity of microscopic examination of wet mount with the potassium hydroxide (KOH) was 61% and 70% with Gram's stain when using the culture of vaginal fluid as gold standard for diagnosis of candidiasis . Among the 44 positives cultures, isolated species of yeast from vaginal swabs were C . albicans (59%), C . tropicalis (23%), C . glabrata (14%) and C . krusei (4%) . This study reports the first characterization of 26 C . albicans stocks from Nicaragua by the random amplified polymorphic DNA method . The genetic analysis in this small C . albicans population showed the existence of linkage disequilibrium, which is consistent with the hypothesis that C . albicans undergoes a clonal propagation.

Cell Immunol, 2002 Jul-Aug, 218(1-2), 46 - 58
Candida albicans infection enhances immunosuppression induced by cyclophosphamide by selective priming of suppressive myeloid progenitors for NO production; Angulo I et al.; Systemic infections caused by fungi after cytoreductive therapies are especially difficult to deal with in spite of currently available antimicrobials . However, little is known about the effects of fungi on the immune system of immunosuppressed hosts . We have addressed this by studying the in vitro T cell responses after systemic infection with Candida albicans in cyclophosphamide-treated mice . After cyclophosphamide treatment, a massive splenic colonization of the spleens, but not lymph nodes, by immature myeloid progenitor (Ly-6G(+)CD11b(+))cells is observed . These cells are able to suppress proliferation of T lymphocytes via a nitric oxide (NO)-dependent mechanism . Systemic infection with a sublethal dose of C . albicans did not cause immunosuppression per se but strongly increased NO-dependent suppression in cyclophosphamide-treated mice, by selective priming of suppressive myeloid progenitors (Ly-6G(+)CD11b(+)CD31(+)CD40(+)WGA(+)CD117(low/-)CD34(low/-)) for iNOS protein expression . The results indicate that systemic C . albicans infection can augment the effects of immunosuppressive therapies by promoting functional changes in immunosuppressive cells.

J Endod, 2002 Nov, 28(11), 770 - 3
Fungal infection of the radicular dentin; Siqueira JF Jr et al.; Although fungi have been detected in infected root canals, their precise role as endodontic pathogens has not yet been elucidated . The purpose of this study was to investigate the pattern of radicular dentin colonization by five fungal species . Bovine root sections were infected with each of the following fungal species: Candida albicans, Candida glabrata, Candida guilliermondii, Candida parapsilosis, and Saccharomyces cerevisiae . After 14 days, the sections were fixed in glutaraldehyde, split into two halves, critical point-dried in CO2, sputter-coated with gold, and examined under scanning electron microscopy . Regardless of the species, single or budding yeast cells were the only fungal forms observed . C . albicans colonized most of the specimens . On the other hand, the other four fungal species presented discrete or no colonization of the radicular dentin . C . albicans showed different patterns of dentin infection . In some specimens, colonization of the dentinal surface was slight and no penetration within dentinal tubules was observed . In the other specimens, some areas of the root canal walls were covered with large colonies of yeast cells and some dentinal tubules were heavily infected . The results suggested that whereas C . albicans showed the ability to colonize dentin, the other four fungal species did not . This can help to explain why C . albicans is the fungal species most often found in endodontic infections.

J Biol Chem, 2003 Feb 21, 278(8), 5523 - 30 Epub 2002 Dec 04.
Synthesis and characterization of the first potent inhibitor of yapsin 1 . Implications for the study of yapsin-like enzymes; Cawley NX et al.; The potent peptidic inhibitor, Y1, of the basic residue-specific yeast aspartyl protease, yapsin 1, was synthesized and characterized . The inhibitor was based on the peptide sequence of a cholecystokinin(13-33) analog that yapsin 1 cleaved with an efficiency of 5.2 x 10(5) m(-1) s(-1) (Olsen, V., Guruprasad, K., Cawley, N . X., Chen, H . C., Blundell, T . L., and Loh, Y . P . (1998) Biochemistry 37, 2768-2777) . The apparent K(i) of Y1 for the inhibition of yapsin 1 was determined to be 64.5 nm, and the mechanism is competitive . Y2 was also developed as an analog of Y1 for coupling to agarose beads . The resulting inhibitor-coupled agarose beads were successfully used to purify yapsin 1 to apparent homogeneity from conditioned medium of a yeast expression system . Utilization of this new reagent greatly facilitates the purification of yapsin 1 and should also enable the identification of new yapsin-like enzymes from mammalian and nonmammalian sources . In this regard, Y1 also efficiently inhibited Sap9p, a secreted aspartyl protease from the human pathogen, Candida albicans, which has specificity for basic residues similar to yapsin 1 and might provide the basis for the prevention or control of its virulence . A single-step purification of Sap9p from conditioned medium was also accomplished with the inhibitor column . N-terminal amino acid sequence analysis yielded two sequences indicating that Sap9p is composed of two subunits, designated here as alpha and beta, similar to yapsin 1.

Bioorg Med Chem Lett, 2003 Jan 6, 13(1), 87 - 91
Design and synthesis of novel benzofurans as a new class of antifungal agents targeting fungal N-myristoyltransferase . Part 3; Kawasaki K et al.; A new series of acid-stable antifungal agents having strong inhibitory activity against Candida albicans N-myristoyltransferase (CaNmt) has been developed starting from acid-unstable benzofuranylmethyl aryl ether 2 . The inhibitor design is based on X-ray crystallographic analysis of a CaNmt complex with aryl ether 3 . Among the new inhibitors, pyridine derivative 8b and benzimidazole derivative 8k showed clear antifungal activity in a murine systemic candidiasis model.

J Infect Dis, 2001 Dec 1, 184(11), 1489 - 93
Growth inhibition of Candida albicans by human vaginal epithelial cells; Barousse MM et al.; Vulvovaginal candidiasis (VVC) is a common mucosal infection caused by Candida species in women of childbearing age . Although acute VVC affects a large number of women and is often precipitated by hormonal fluctuations involving high estrogen levels, recurrent VVC (RVVC) affects another 5%-10% of women without any known predisposing factors . We have recently reported that vaginal epithelial cells from nonhuman primates and mice inhibit the growth of Candida albicans in vitro, which may represent an innate host defense mechanism against C . albicans at the vaginal mucosa . In the present study, we show that vaginal epithelial cells collected from healthy women with no history of VVC also exhibit anti-Candida activity, with no differences in activity at various stages of the menstrual cycle . Women diagnosed with RVVC, on the other hand, have reduced epithelial cell anti-Candida activity . These results are further evidence that vaginal epithelial cells provide an innate host resistance mechanism against Candida and that reduced activity may contribute to RVVC.

Cell Microbiol, 2002 Dec, 4(12), 805 - 12
Endocytosis of Candida albicans by vascular endothelial cells is associated with tyrosine phosphorylation of specific host cell proteins; Belanger PH et al.; Candida albicans escapes from the bloodstream by invading the endothelial cell lining of the vasculature . In vitro, C . albicans invades endothelial cells by inducing its own endocytosis . We examined whether this process is regulated by the tyrosine phosphorylation of endothelial cell proteins . We found that endocytosis of wild-type C . albicans was accompanied by the tyrosine phosphorylation of two endothelial cell proteins with molecular masses of 80 and 82 kDa . The phosphorylation of these proteins was closely associated with the endocytosis of C . albicans because these proteins were phosphorylated in response to the endocytosis of both live and killed organisms, but they were not phosphorylated in endothelial cells infected with a poorly endocytosed strain of C . albicans . The tyrosine kinase inhibitors genistein and tyrphostin 47 blocked the phosphorylation of the two endothelial cell proteins and significantly reduced endocytosis of C . albicans . Therefore, C . albicans probably induces its own endocytosis by stimulating the tyrosine phosphorylation of two endothelial cell proteins.

Med Mycol, 2002 Oct, 40(5), 485 - 92
Phenotypic and functional changes on phagocytic cells recruited at the site of Candida albicans infection after chronic varied stress exposure; Rodriguez-Galan MC et al.; The transition of Candida albicans from commensalism to pathogenicity is associated with the immune status of the host; resistance to fungus involves macrophages (Mphi) and polymorphonuclear neutrophils (PMN), which act as effector cells . T-cell function is also involved . Previously, we found that in Wistar rats exposed to chronic varied stress (CVS) immediately after C . albicans infection (Ca-S group) some functions of phagocytic cells, such as killer activity and NO production, were strongly modified compared with unstressed, infected animals (Ca group) . We examined the phenotypic and functional changes of these effector cells recruited at the site of C . albicans infection . The recruitment of peritoneal cells (PC) was markedly reduced in Ca-S animals and the arrival of Mphi and PMN was selectively diminished after CVS exposure . The integrin CD11b/CD18, implicated in migration and C . albicans phagocytosis, was downregulated in Mphi of Ca-S animals . The activation markers CD54 and MHC-II were upregulated in Mphi after fungal contact . The expression of CD54 was only changed in Ca-S rats . Finally, TNF-alpha production was reduced in PC of Ca-S animals, suggesting an impairment of functional activity . Taken together, the phenotypic and functional changes detected in effector cells may account for the decreased resistance to candidiasis seen in conjunction with CVS . The changes seen also expand our knowledge of the role of Mphi in the control of C . albicans dissemination.

Med Mycol, 2002 Oct, 40(5), 471 - 8
Gene expression of 70 kDa heat shock protein of Candida albicans: transcriptional activation and response to heat shock; Sandini S et al.; CaHSP70 (70 kDa heat shock protein) is a highly immunogenic protein of Candida albicans . We have studied heat shock-induced expression of the CaHSP70 gene under germ tube-inductive and non-inductive conditions . The CaHSP70 upstream regulatory region was cloned and sequenced . It contains at least three heat shock elements (HSEs), specific DNA sequences that are bound by the heat shock transcription factor (HSF), and one stress response element (STRE), which is an upstream activator sequence (UAS) that causes transcription activation under stress . The binding of HSF to HSE in the CaHSP70 promoter region is constitutive, although the mobility of protein/DNA complexes is altered after heat shock . The CaHSP70 promoter was cloned into a lacZ reporter plasmid, and was able to respond to heat shock in C . albicans as well as in Saccharomyces cerevisiae.

Med Mycol, 2002 Oct, 40(5), 465 - 9
Inefficient delivery of yeast cells as an explanation for reduced plating efficiency of Candida albicans; Bhatti MA et al.; The plating efficiency for fungal yeast cells is usually less than that expected from microscopic counts, and a number of explanations for this phenomenon have been proposed . The present study was undertaken to explore possible reasons for reduced plating efficiency of Candida albicans . Explanations that we evaluated and found unlikely included: ineffectiveness of different culture media and/or incubation temperatures for growing colonies, insufficient area of the plate available for expression of individual colonies, production of microcolonies, and inaccurate counting of the organisms in the inoculum . An assay for delivery of the inoculum into tissue culture plate wells indicated that reduced delivery of the organisms accounted for lower than expected plating efficiency . C . albicans yeast cells grown under low glucose conditions and expected to have reduced adhesiveness were found to have higher values for both delivery and plating efficiency in our assays . In summary, our results indicate that reduced plating efficiency for C . albicans under the conditions used for these experiments is best explained by the loss of some yeast cells during preparation of the inocula or delivery of the yeast cells onto the plates.

J Antimicrob Chemother, 2002 Dec, 50(6), 999 - 1002
Point prevalence, microbiology and fluconazole susceptibility patterns of yeast isolates colonizing the oral cavities of HIV-infected patients in the era of highly active antiretroviral therapy; Barchiesi F et al.; We conducted a prospective study to address the prevalence and microbiological characteristics of yeast isolates colonizing the oral cavities of HIV-infected patients undergoing highly active antiretroviral therapy . Sixty-eight patients (67%) from a total of 102 were found to be colonized with yeasts . Sixty-five patients carried a single species (60 Candida albicans, three Candida glabrata and two Candida krusei) and three patients had mixed colonization of C . albicans and C . krusei . The status of yeast carrier was not associated with the number of CD4 cells or the viral load . Similarly, the type of antiretroviral regimen was not associated with the carriage of Candida spp . The only predictor of Candida colonization was a previous history of oropharyngeal candidiasis (P = 0.009) . Although many patients in this series had already been treated with repeated courses of fluconazole therapy for previous episodes of oropharyngeal candidiasis, fluconazole susceptibility patterns showed that 93% of yeasts were susceptible to this triazole in vitro (MIC < or = 8.0 mg/L).

Bull Exp Biol Med, 2002 Jul, 134(1), 103 - 5
Modified method for evaluation of plasma membrane integrity in eukaryotic cell; Arzumanyan VG et al.; We propose a method for evaluation of the number of viable cells by the content of bromcresol purple dye absorbed by dead cells from the incubation medium . Myramistin was used as a pore-forming agent . The number of live cells in yeast suspension inversely correlated with the percentage of dye absorbed by cells . The method is simple and requires no special equipment . The effect of myramistin on Candida albicans and Malassezia sympodialis cells and on epitheliocytes was evaluated . Two-hour treatment with myramistin in a concentration commonly used in clinical practice (100 mug/ml) decreased the number of viable cells by 2 and 1 order of magnitude, respectively . Epitheliocytes under the same conditions absorbed approximately the same mount of the dye as Candida cells.

Nat Rev Genet, 2002 Dec, 3(12), 918 - 30
Candida Albicans: a molecular revolution built on lessons from budding yeast; Berman J et al.; Candida albicans is an opportunistic fungal pathogen that is found in the normal gastrointestinal flora of most healthy humans . However, in immunocompromised patients, blood-stream infections often cause death, despite the use of anti-fungal therapies . The recent completion of the C . albicans genome sequence, the availability of whole-genome microarrays and the development of tools for rapid molecular-genetic manipulations of the C . albicans genome are generating an explosion of information about the intriguing biology of this pathogen and about its mechanisms of virulence . They also reveal the extent of similarities and differences between C . albicans and its benign relative, Saccharomyces cerevisiae.

Hum Exp Toxicol, 2002 Sep-Oct, 21(9-10), 507 - 12
Practical aspects of including functional endpoints in developmental toxicity studies . Case study: immune function in HuCD4 transgenic mice exposed to anti-CD4 MAb in utero; Herzyk DJ et al.; Keliximab is a human-cynomolgus monkey chimeric (Primatized) monoclonal antibody with specificity for human and chimpanzee CD4 . As the preclinical safety assessment of biopharmaceuticals requires evaluation in pharmacologically responsive species, comprehensive toxicology studies, including reproductive toxicity, of this antibody were conducted in a human CD4 transgenic mouse model . The reproductive toxicology studies included a pre- and postnatal development study that incorporated immunotoxicological evaluation of offspring (F1) mice . The potential effects of exposure to treating maternal mice (F0) with keliximab during pregnancy and lactation on offspring viability, physical growth, neurobehavioral development, reproductive function, lymphoid tissue morphological structure, lymphocyte subsets and host resistance to Candida albicans infection were assessed . The results showed no impairment of these functions . The use of F1 transgenic mice in study with keliximab provides an example of a novel practical approach to assess developmental immunotoxicity within a study of pre- and postnatal development designed in accordance with ICH Guidelines.

Turk J Pediatr, 2002 Oct-Dec, 44(4), 339 - 42
Neonatal onset propionic acidemia without acidosis: a case report; Akman I et al.; Propionic acidemia is an inherited disorder of organic acid metabolism characterized by a spectrum of clinical and biochemical findings . The usual presentation is life-threatening ketoacidosis and hyperammonemia . In this report we present a neonate with propionic acidemia presenting with prominent neurologic problems without ketoacidosis . The patient had a serum ammonia level of 3,500 microg/dl which was effectively lowered to normal values in 48 hours by peritoneal dialysis, with remarkable improvement in neurologic status . However, she developed Candida albicans peritonitis, and sepsis and died of cardiorespiratory failure . Infants who have an early onset propionic acidemia have a high mortality and morbidity rate . In conclusion, propionic acidemia should be in the differential diagnosis of patients with neurologic symptoms and hyperammonemia with or without acidosis.

Int J Antimicrob Agents, 2002 Dec, 20(6), 472 - 5
Epidemiology of clinical isolates of Candida albicans and their susceptibility to triazoles; Wroblewska MM et al.; The study comprised strains of Candida albicans isolated from patients hospitalised in a tertiary care hospital during a 2-year period . In total 851 strains were cultured, including 379 (44.5%) strains from internal medicine patients, 243 (28.6%) from surgical patients and 229 (26.9%) from patients in the surgical intensive care unit . The strains were tested for susceptibility to the triazoles: fluconazole and itraconazole . There were 523 (61.5%) strains susceptible, 11 strains (1.3%) showed intermediate susceptibility and 317 (37.2%) were resistant to fluconazole, while 403 (47.3%) strains were susceptible, 43 (5.1%) intermediately susceptible and 405 (47.6%) resistant to itraconazole . Regular surveillance of fungal resistance patterns should be carried out and there should be prudent use of hospital triazole usage .

Int J Antimicrob Agents, 2002 Dec, 20(6), 468 - 71
Evaluation of the in vitro activity of caspofungin against bloodstream isolates of Candida species from cancer patients: comparison of Etest and NCCLS reference methods; Laverdiere M et al.; The in vitro activity of caspofungin (CAS) was compared with the activity of fluconazole, itraconazole and amphotericin B against 178 bloodstream Candida spp . from cancer patients . The activities were assessed using the reference NCCLS M-27A microdilution method and the Etest method . With both the NCCLS microdilution reference method and the Etest method, CAS was the most active agent (MIC90s 0.19-0.5 mg/l) against Candida albicans, C . glabrata and C . tropicalis . CAS showed substantial activity against azole-resistant Candida . The percentages of agreement within +/-2 dilutions between the NCCLS reference microdilution method and Etest MICs ranged from 81 to 97% . CAS showed good in vitro activity against invasive azole-susceptible and azole-resistant Candida isolates . The CAS Etest MICs correlated well with the NCCLS reference MICs and may provide more choice for laboratories in assessing the activity of antifungal agents . The clinical correlation of these in vitro observations needs to be established .

Diagn Microbiol Infect Dis, 2002 Oct, 44(2), 169 - 74
Antifungal susceptibility of South African oral yeast isolates from HIV/AIDS patients and healthy individuals; Blignaut E et al.; The in vitro antifungal susceptibility profile of 589 oral yeast isolates from HIV/AIDS patients and healthy South Africans was determined against amphotericin B, nystatin, 5-fluorocytosine (5-FC), clotrimazole, miconazole, ketoconazole, itraconazole and fluconazole . The broth microdilution method of the National Committee on Clinical Laboratory Standards was used and MIC(50) and MIC(90) determined . A 100% susceptibility to fluconazole was observed among the 466 isolates of Candida albicans . Among C . krusei, the second most common isolate, only 2.6% of isolates were susceptible to fluconazole and itraconazole . Despite the lack of previous exposure to antifungal agents, very little difference was observed in the antifungal profile between the South African isolates and isolates from the United States (U.S.), Canada and South America . South Africa has a particularly high incidence of HIV-infection and oral candidiasis is the most common oral complication in these patients . This study provides important baseline data as the isolates were collected prior to fluconazole being made freely available to HIV/AIDS patients attending government health clinics.

Diagn Microbiol Infect Dis, 2002 Oct, 44(2), 163 - 7
Frequency and clinical significance of bloodstream infections caused by C albicans strains with reduced susceptibility to fluconazole; Munoz P et al.; Reduced susceptibility to fluconazole (RSF) is relatively common in non-albicans Candida isolates and in Candida albicans recovered from HIV-infected patients with relapsing Candida stomatitis or esophagitis . However, little clinical data on bloodstream infections caused by C . albicans with RSF is available . We analyzed 116 episodes of C . albicans fungemia detected over an 11-year period . Four patients (3.4%) had a blood isolate of C . albicans with RSF . Fluconazole MICs were 16 (3 SDD strains) and 128 microg/ml (1 resistant strain), respectively . Three of the patients were HIV (+) and the fourth was a liver transplant recipient . All of them had been previously treated with an azole compound . The liver recipient had breakthrough fungemia while being treated with 400 mg of preemptive fluconazole despite having an MIC of 16 microg/ml . Fluconazole clinical failure was documented in two of the remaining three cases . Only five other patients with C . albicans fungemia caused by fluconazole-resistant strains (>or=64 microg/ml) are described in the literature . Candida albicans fungemia produced by strains with RSF is still uncommon . It should be suspected in patients previously treated with azole agents or with breakthrough fungemia . In our experience, fluconazole remains a safe option for the treatment of most C . albicans fungemias, although surveillance seems advisable.

Contemp Top Lab Anim Sci, 1998 May, 37(3), 43 - 46
Efficacy of Alkaline Hydrolysis as an Alternative Method for Treatment and Disposal of Infectious Animal Waste; Kaye G et al.; The efficacy of alkaline hydrolysis as an alternative for incineration or autoclaving during treatment and disposal of infectious waste was evaluated by testing for the destruction of samples of pure cultures of selected infectious microorganisms during digestion of 114 to 136-kg loads of animal carcasses in an animal tissue digestor at the Albany Medical College . Ten milliliter samples of pure cultures of each microorganism were divided among 3 dialysis bags made from narrow diameter dialysis tubing, and each of these bags was placed inside another dialysis bag made from larger diameter dialysis tubing . Each double-bagged sample was suspended from the cover of the carcass basket of the tissue digestor so that it was completely covered by hot alkaline digestion solution during the carcass digestion process . The following organisms were required by the New York State Department of Health as representative pathogens for testing sterilization capabilities of the procedure: Staphylococcus aureus, Mycobacterium fortuitum, Candida albicans, Bacillus subtilis, Pseudomonas aeruginosa, Aspergillus fumigatus, Mycobacterium bovis BCG, MS-2 bacteriophage, and Giardia muris . Animal carcasses included pigs, sheep, rabbits, dogs, rats, mice, and guinea pigs . The tissue digestor was operated at 110 to 120 C and approximately 15 lb/in2 (gauge) for 18 h before the system was allowed to cool to 50 C and dialysis bags were retrieved and submitted for microbial culture . None of the samples obtained from the dialysis bags after the digestion process yielded indicator bacteria or yeast . Giardia cysts were completely destroyed; only small fragments of what appeared to be cyst wall could be recognized with light microscopic examination . No plaque-forming units were detected with MS-2 bacteriophage after digestion . Samples of the hydrolyzate also did not yield growth on culture media . Animal carcasses were completely solubilized and digested, with only the inorganic components of the bones and teeth remaining after draining and rinsing of the digestion vessel . Alkaline hydrolysis, as performed in this tissue digestor, completely destroyed all representative classes of potentially infectious agents as well as disposing of animal carcasses by solubilization and digestion.

Eukaryot Cell, 2002 Aug, 1(4), 514 - 25
Large-scale identification of putative exported proteins in Candida albicans by genetic selection; Monteoliva L et al.; In all living organisms, secreted proteins play essential roles in different processes . Of special interest is the construction of the fungal cell wall, since this structure is absent from mammalian cells . The identification of the proteins involved in its biogenesis is therefore a primary goal in antifungal research . To perform a systematic identification of such proteins in Candida albicans, we carried out a genetic screening in which in-frame fusions with an intracellular allele of invertase gene SUC2 of Saccharomyces cerevisiae can be used to select and identify putatively exported proteins in the heterologous host S . cerevisiae . Eighty-three clones were selected, including 11 previously identified genes from C . albicans as well as 41 C . albicans genes that encode proteins homologous to already described proteins from related organisms . They include enzymes involved in cell wall synthesis and protein secretion . We also found membrane receptors and transporters presumably related to the interaction of C . albicans with the environment as well as extracellular enzymes and proteins involved in different morphological transitions . In addition, 11 C . albicans open reading frames (ORFs) identified in this screening encode proteins homologous to unknown or putative proteins, while 5 ORFs encode novel secreted proteins without known homologues in other organisms . This screening procedure therefore not only identifies a set of targets of interest in antifungal research but also provides new clues for understanding the topological locations of many proteins involved in processes relevant to the pathogenicity of this microorganism.

Eukaryot Cell, 2002 Jun, 1(3), 420 - 31
The Golgi GDPase of the fungal pathogen Candida albicans affects morphogenesis, glycosylation, and cell wall properties; Herrero AB et al.; Cell wall mannoproteins are largely responsible for the adhesive properties and immunomodulation ability of the fungal pathogen Candida albicans . The outer chain extension of yeast mannoproteins occurs in the lumen of the Golgi apparatus . GDP-mannose must first be transported from the cytosol into the Golgi lumen, where mannose is transferred to mannans . GDP is hydrolyzed by a GDPase, encoded by GDA1, to GMP, which then exits the Golgi lumen in a coupled, equimolar exchange with cytosolic GDP-mannose . We isolated and disrupted the C . albicans homologue of the Saccharomyces cerevisiae GDA1 gene in order to investigate its role in protein mannosylation and pathogenesis . CaGda1p shares four apyrase conserved regions with other nucleoside diphosphatases . Membranes prepared from the C . albicans disrupted gda1/gda1 strain had a 90% decrease in the ability to hydrolyze GDP compared to wild type . The gda1/gda1 mutants showed a severe defect in O-mannosylation and reduced cell wall phosphate content . Other cell wall-related phenotypes are present, such as elevated chitin levels and increased susceptibility to attack by beta-1,3-glucanases . Our results show that the C . albicans organism contains beta-mannose at their nonreducing end, differing from S . cerevisiae, which has only alpha-linked mannose residues in its O-glycans . Mutants lacking both alleles of GDA1 grow at the same rate as the wild type but are partially blocked in hyphal formation in Lee solid medium and during induction in liquid by changes in temperature and pH . However, the mutants still form normal hyphae in the presence of serum and N-acetylglucosamine and do not change their adherence to HeLa cells . Taken together, our data are in agreement with the hypothesis that several pathways regulate the yeast-hypha transition . Gda1/gda1 cells offer a model for discriminating among them.

Eukaryot Cell, 2002 Jun, 1(3), 353 - 65
Roles of TUP1 in switching, phase maintenance, and phase-specific gene expression in Candida albicans; Zhao R et al.; Candida albicans strain WO-1 switches spontaneously and reversibly between a "white" and "opaque" phenotype that affects colony morphology, cellular phenotype, and expression of a number of phase-specific genes and virulence traits . To assess the role of the transcription regulator Tup1p in this phenotypic transition, both TUP1 alleles were deleted in the mutant delta tup1 . Delta tup1 formed "fuzzy large" colonies made up of cells growing exclusively in the filamentous form . Delta tup1 cells did not undergo the white-opaque transition, but it did switch spontaneously, at high frequency (approximately 10(-3)), and unidirectionally through the following sequence of colony (and cellular) phenotypes: "fuzzy large" (primarily hyphae) --> "fuzzy small" (primarily pseudohyphae) --> "smooth" (primarily budding yeast) --> "revertant fuzzy" (primarily pseudohyphae) . Northern analysis of white-phase, opaque-phase, and hypha-associated genes demonstrated that Tup1p also plays a role in the regulation of select phase-specific genes and that each variant in the delta tup1 switching lineage differs in the level of expression of one or more phase-specific and/or hypha-associated genes . Using a rescued delta tup1 strain, in which TUP1 was placed under the regulation of the inducible MET3 promoter, white- and opaque-phase cells were individually subjected to a regime in which TUP1 was first downregulated and then upregulated . The results of this experiment demonstrated that (i) downregulation of TUP1 led to exclusive filamentous growth in both originally white- and opaque-phase cells; (ii) the white-phase-specific gene WH11 continued to be expressed in TUP1 downregulated cultures originating from white-phase cells, while WH11 expression remained repressed in TUP1-downregulated cultures originating from opaque-phase cells, suggesting that cells maintained phase identity in the absence of TUP1 expression; and (iii) subsequent upregulation of TUP1 resulted in mass conversion of originally white-phase cells to the opaque phase and maintenance of originally opaque-phase cells in the opaque phase and in the resumption in both cases of switching, suggesting that TUP1 reexpression turns on the switching system in the opaque phase.

Eukaryot Cell, 2002 Feb, 1(1), 95 - 104
CDC42 is required for polarized growth in human pathogen Candida albicans; Ushinsky SC et al.; Cdc42p is a member of the RAS superfamily of GTPases and plays an essential role in polarized growth in many eukaryotic cells . We cloned the Candida albicans CaCDC42 by functional complementation in Saccharomyces cerevisiae and analyzed its function in C . albicans . A double deletion of CaCDC42 was made in a C . albicans strain containing CaCDC42 under the control of the PCK1 promoter . When expression of the heterologous copy of CaCDC42 was repressed in this strain, the cells ceased proliferation . These arrested cells were large, round, and unbudded and contained predominantly two nuclei . The PCK1-mediated overexpression of wild-type CaCdc42p had no effect on cells . However, in cells overexpressing CaCdc42p containing the dominant-negative D118A substitution, proliferation was blocked and the arrested cells were large, round, unbudded, and multinucleated, similar to the phenotype of the cdc42 double-deletion strain . Cells overexpressing CaCdc42p containing the hyperactive G12V substitution also ceased proliferation in yeast growth medium; in this case the arrested cells were multinucleated and multibudded . An intact CAAX box is essential for the phenotypes associated with either CaCdc42p(G12V) or CaCdc42p(D118A) ectopic expression, suggesting that membrane attachment is involved in CaCdc42p function . In addition, the lethality caused by ectopic expression of CaCdc42p(G12V) was suppressed by deletion of CST20 but not by deletion of CaCLA4 . CaCdc42p function was also examined under hypha-inducing conditions . Cdc42p depletion prior to hyphal induction trapped cells in a round, unbudded state, while depletion triggered at the same time as hyphal induction permitted the initiation of germ tubes that failed to be extended . Ectopic expression of either the G12V or D118A substitution protein modified hyphal formation in a CAAX box-dependent manner . Thus, CaCdc42p function appears important for polarized growth of both the yeast and hyphal forms of C . albicans.

Eukaryot Cell, 2002 Apr, 1(2), 241 - 8
Aspergillus nidulans swoF encodes an N-myristoyl transferase; Shaw BD et al.; Polar growth is a fundamental process in filamentous fungi and is necessary for disease initiation in many pathogenic systems . Previously, swoF was identified in Aspergillus nidulans as a single-locus, temperature-sensitive (ts) mutant aberrant in both polarity establishment and polarity maintenance . The swoF gene was cloned by complementation of the ts phenotype and sequenced . The derived protein sequence had high identity with N-myristoyl transferases (NMTs) found in fungi, plants, and animals . In addition, wild-type growth at restrictive temperature was partially restored by the addition of myristic acid to the growth medium . Sequencing revealed that the mutation in swoF changes the conserved aspartic acid 369 to a tyrosine . The predicted A . nidulans SwoF protein, SwoFp, was homology modeled based on crystal structures of NMTs from Saccharomyces cerevisiae and Candida albicans . The D369Y swoF mutation is on the opposite face of the protein, distal to the myristoyl coenzyme A and peptide substrate binding sites . In wild-type NMTs, D369 appears to stabilize a structural beta-strand bend through two hydrogen bonds and an ionic interaction . These stabilizing bonds are abolished in the D369Y mutant . We hypothesize that a substrate of SwoFp must be myristoylated for proper polarity establishment and maintenance . The mutation prevents the proper function of SwoFp at restrictive temperature and thus blocks polar growth.

Eukaryot Cell, 2002 Apr, 1(2), 213 - 28
Myosin I is required for hypha formation in Candida albicans; Oberholzer U et al.; The pathogenic yeast Candida albicans can undergo a dramatic change in morphology from round yeast cells to long filamentous cells called hyphae . We have cloned the CaMYO5 gene encoding the only myosin I in C . albicans . A strain with a deletion of both copies of CaMYO5 is viable but cannot form hyphae under all hypha-inducing conditions tested . This mutant exhibits a higher frequency of random budding and a depolarized distribution of cortical actin patches relative to the wild-type strain . We found that polar budding, polarized localization of cortical actin patches, and hypha formation are dependent on a specific phosphorylation site on myosin I, called the "TEDS-rule" site . Mutation of this serine 366 to alanine gives rise to the null mutant phenotype, while a S366D mutation, the product of which mimics a phosphorylated serine, allows hypha formation . However, the S366D mutation still causes a depolarized distribution of cortical actin patches in budding cells, similar to that in the null mutant . The localization of CaMyo5-GFP together with cortical actin patches at the bud and hyphal tips is also dependent on serine 366 . Intriguingly, the cortical actin patches in the majority of the hyphae of the mutant expressing Camyo5(S366D) were depolarized, suggesting that although their distribution is dependent on myosin I localization, polarized cortical actin patches may not be required for hypha formation.

Res Microbiol, 2002 Nov, 153(9), 611 - 20
Cloning and characterization of PRB1, a Candida albicans gene encoding a putative novel endoprotease B and factors affecting its expression; Orozco I et al.; Several cDNA fragments corresponding to transcripts differentially expressed under conditions that favor mycelial growth of Candida albicans were identified by the "differential display" technique . One of these was cloned and used as a probe to rescue the full gene from a genomic library of the fungus . The sequence identified a single, uninterrupted open reading frame of 1395 nucleotides encoding a putative protein of 465 residues and a theoretical molecular weight of 50.3 kDa, present in the genome as a single copy located at chromosome 2 in different strains . The gene product showed high homology with subtilisin-like proteases, mainly PRB1, the vacuolar B protease from Saccharomyces cerevisiae, and for this reason it was designated as a putative CaPRB1 . Expression of the gene was not directly related to fungal morphogenesis, but to the initial response to inducers: Heat shock and the presence of N-acetyl glucosamine . It was also subject to nitrogen, but not to carbon catabolite repression, although glucose inhibited the GlcNAc stimulatory effect . The gene was, in our hands, unable to complement PRB1 mutation in S . cerevisiae . C . albicans prb null mutants did not show any distinct alteration in the phenotype . CaPRB1 is the first gene coding for a putative vacuolar serine protease cloned from C . albicans.

Ann Surg, 2002 Dec, 236(6), 759 - 66; disscussion 767
Laparoscopic versus open colorectal surgery: a randomized trial on short-term outcome; Braga M et al.; OBJECTIVE: The primary endpoint was to compare the impact of laparoscopic and open colorectal surgery on 30-day postoperative morbidity . Lymphocyte proliferation to mitogens and gut oxygen tension were surrogate endpoints . SUMMARY BACKGROUND DATA: Evidence-based proof of the effect of laparoscopic colorectal surgery on immunometabolic response and clinically relevant outcome variables is scanty . Further randomized trials are desirable before proposing laparoscopy as a superior technique . METHODS: Two hundred sixty-nine patients with colorectal disease were randomly assigned to laparoscopic (n = 136) or open (n = 133) colorectal resection . Four trained members of the surgical staff who were not involved in the study registered postoperative complications . Lymphocyte proliferation to Candida albicans and phytohemagglutinin was evaluated before and 3 and 15 days after surgery . Operative gut oxygen tension was monitored continuously by a polarographic microprobe . RESULTS: In the laparoscopic group the conversion rate was 5.1% . The overall morbidity rate was 20.6% in the laparoscopic group and 38.3% in the open group . Postoperative infections occurred in 15 of the 136 patients in the laparoscopic group and 31 of the 133 patients in the open group . The mean length of hospital stay was 10.4 +/- 2.9 days in the laparoscopic group and 12.5 +/- 4.1 days in the open group . On postoperative day 3, lymphocyte proliferation was impaired in both groups . Fifteen days after surgery, the proliferation index returned to baseline values only in the laparoscopic group . Intraoperative gut oxygen tension was higher in the laparoscopic than in the open group . CONCLUSIONS: Laparoscopic colorectal surgery resulted in a significant reduction of 30-day postoperative morbidity . Lymphocyte proliferation and gut oxygen tension were better preserved in the laparoscopic group than in the open group.

Nephrol Dial Transplant, 2002 Dec, 17(12), 2189 - 95
Superior antimicrobial activity of trisodium citrate over heparin for catheter locking; Weijmer MC et al.; BACKGROUND: Haemodialysis catheters used for vascular access are frequently complicated by infection and catheter-related thrombosis . Improvement of interdialytic locking solutions could reduce these problems . Trisodium citrate (TSC) has been advocated in recent years because it might have antimicrobial qualities . METHODS: Antimicrobial efficacy of four concentrations of TSC (2.2, 7.5, 15 and 30%) was compared with three equi-osmolal sodium chloride (NaCl) concentrations, unfractionated heparin 5000 IU/ml and a solution of gentamicin 1 mg/ml in TSC 7.5% . We analysed antimicrobial properties by two classical in vitro susceptibility tests . All tests were performed in triplicate by incubation of test fluids with Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans . RESULTS: Increasing TSC concentrations effectively killed the staphylococcal strains in both assays . For E.coli and P.aeruginosa complete killing was achieved only with TSC 30% . TSC 30% was also the only solution that significantly inhibited growth of C.albicans . Heparin manifested no antimicrobial effect of any significance . Adding gentamicin to TSC provided superior bacterial growth inhibition but had no effect on yeast growth . TSC solutions manifested superior antimicrobial activity compared with iso-osmolal NaCl solutions in both assays . CONCLUSION: This in vitro study demonstrates superior antimicrobial activity of TSC, especially in higher concentrations, in contrast to heparin . The mechanism seems to differ from hyperosmolality . Ca(2+) and Mg(2+) chelating effects are probably more important . Adding gentamicin provided the most potent antimicrobial solution . However, for reasons concerning development of bacterial resistance and sensitization of the patient, continuous exposition to aminoglycosides seems not advisable.

J Clin Microbiol, 2002 Dec, 40(12), 4479 - 85
High oral prevalence of Candida krusei in leprosy patients in northern Thailand; Reichart PA et al.; Although Candida albicans is the most common human yeast pathogen, other Candida species such as C . krusei are now recognized as emerging agents, especially in patients with human immunodeficiency virus (HIV) disease . C . krusei is inherently resistant to the widely used triazole antifungal fluconazole and poses therapeutic problems, especially in systemic candidiasis . In a surveillance study of leprosy patients (with arrested or burnt-out disease) in a leprosarium in northern Thailand, we found a rate of oral carriage of C . krusei (36%) significantly (P < 0.05) higher than that for a healthy control group (10%) . Among the Candida-positive patients, 16 of 35 (46%) carried C . krusei, while C . albicans was the second most common isolate (12 of 35 patients; 34%) . The corresponding figures for the control group were 2 of 13 (15%) and 6 of 13 (46%), respectively . Studies of the antifungal resistance of the C . krusei isolates from patients indicated that all except one of the isolates were resistant to fluconazole, two isolates were resistant to ketoconazole, and all isolates were sensitive to amphotericin B . Evaluation of their genetic profiles by randomly amplified polymorphic DNA analysis with three different primers and subsequent analysis of the gel profiles by computerized cluster-derived dendrograms revealed that the C . krusei isolates from patients belonged to 10 disparate clusters, despite the origin from a single locale . These nascent findings indicate an alarmingly high prevalence of a Candida species resistant to a widely used antifungal in a part of the world where HIV disease is endemic.

Mol Microbiol, 2002 Dec, 46(5), 1345 - 51
Many of the genes required for mating in Saccharomyces cerevisiae are also required for mating in Candida albicans; Magee BB et al.; Candida albicans is the single, most frequently isolated human fungal pathogen . As with most fungal pathogens, the factors which contribute to pathogenesis in C . albicans are not known, despite more than a decade of molecular genetic analysis . Candida albicans was thought to be asexual until the discovery of the MTL loci homologous to the mating type (MAT) loci in Saccharomyces cerevisiae led to the demonstration that mating is possible . Using Candida albicans mutants in genes likely to be involved in mating, we analysed the process to determine its similarity to mating in Saccharomyces cerevisiae . We examined disruptions of three of the genes in the MAPK pathway which is involved in filamentous growth in both S . cerevisiae and C . albicans and is known to control pheromone response in the former fungus . Disruptions in HST7 and CPH1 blocked mating in both MTLa and MTL(alpha) strains, whereas disruptions in STE20 had no effect . A disruption in KEX2, a gene involved in processing the S . cerevisiae pheromone Mf(alpha), prevented mating in MTL(alpha) but not MTLa cells, whereas a disruption in HST6, the orthologue of the STE6 gene which encodes an ABC transporter responsible for secretion of the Mfa pheromone, prevented mating in MTLa but not in MTL(alpha) cells . Disruption of two cell wall genes, ALS1 and INT1, had no effect on mating, even though ALS1 was identified by similarity to the S . cerevisiae sexual agglutinin, SAG1 . The results reveal that these two diverged yeasts show a surprising similarity in their mating processes.

Mol Microbiol, 2002 Dec, 46(5), 1335 - 44
A conserved mitogen-activated protein kinase pathway is required for mating in Candida albicans; Chen J et al.; Candida albicans had been thought to lack a mating process until the recent discovery of a mating type-like locus and mating between MTLa and MTL(alpha) strains . To elucidate the molecular mechanisms that regulate mating in C . albicans, we examined the function of Cph1 and its upstream mitogen-activated protein (MAP) kinase pathway in mating, as they are homologues of the pheromone-responsive MAP kinase pathway in Saccharomyces cerevisiae . We found that overexpressing CPH1 in MTLa, but not in MTLa/alpha strains, induced the transcription of orthologues of S . cerevisiae pheromone-induced genes and also increased mating efficiency . Furthermore, cph1 and hst7 mutants were completely defective in mating, and cst20 and cek1 mutants showed reduced mating efficiency, as in S . cerevisiae . The partial mating defect in cek1 results from the presence of a functionally redundant MAP kinase, Cek2 . CEK2 complemented the mating defect of a fus3 kss1 mutant of S . cerevisiae and was expressed only in MTLa or MTL(alpha), but not in MTLa/alpha cell types . Moreover, a cek1 cek2 double mutant was completely defective in mating . Our data suggest that the conserved MAP kinase pathway regulates mating in C . albicans . We also observed that C . albicans mating efficiency was greatly affected by medium composition, indicating the potential involvement of nutrient-sensing pathways in mating in addition to the MAP kinase pathway.

Int J Med Microbiol, 2002 Oct, 292(5-6), 405 - 19
Secreted proteases from pathogenic fungi; Monod M et al.; Many species of human pathogenic fungi secrete proteases in vitro or during the infection process . Secreted endoproteases belong to the aspartic proteases of the pepsin family, serine proteases of the subtilisin family, and metalloproteases of two different families . To these proteases has to be added the non-pepsin-type aspartic protease from Aspergillus niger and a unique chymotrypsin-like protease from Coccidioides immitis . Pathogenic fungi also secrete aminopeptidases, carboxypeptidases and dipeptidyl-peptidases . The function of fungal secreted proteases and their importance in infections vary . It is evident that secreted proteases are important for the virulence of dermatophytes since these fungi grow exclusively in the stratum corneum, nails or hair, which constitutes their sole nitrogen and carbon sources . The aspartic proteases secreted by Candida albicans are involved in the adherence process and penetration of tissues, and in interactions with the immune system of the infected host . For Aspergillus fumigatus, the role of proteolytic activity has not yet been proved . Although the secreted proteases have been intensively investigated as potential virulence factors, knowledge on protease substrate specificities is rather poor and few studies have focused on the research of inhibitors . Knowledge of substrate specificities will increase our understanding about the action of each protease secreted by pathogenic fungi and will help to determine their contribution to virulence.

Int J Med Microbiol, 2002 Oct, 292(5-6), 299 - 311
Co-regulation of pathogenesis with dimorphism and phenotypic switching in Candida albicans, a commensal and a pathogen; Liu H; Candida albicans, a common fungal pathogen of humans, can colonize in many diverse environments of the host and convert between a harmless commensal and a pathogen . Recent advances indicate that C . albicans uses a common set of conserved pathways to regulate dimorphism, mating and phenotypic switching . Major pathways known to regulate dimorphism include a mitogen-activated protein (MAP) kinase pathway through Cph1, the cAMP-dependent protein kinase pathway via Efg1, and Tup1-mediated repression through Rfg1 and Nrg1 . The Cph1-mediated MAP kinase pathway is critical for the mating process, while all three pathways are implicated in the regulation of white-opaque switching . All these developmental pathways regulate the expression of hypha-specific and/or phase-specific genes . A high proportion of hypha-specific genes and phase-specific genes encode proteins that contribute directly or indirectly to pathogenesis and virulence of C . albicans . Therefore, virulence genes are co-regulated with cell morphogenesis . This supports a previous notion that the unique aspects of C . albicans commensalism and pathogenesis may lie in the developmental programs of dimorphism and phenotypic switching.

Biosci Biotechnol Biochem, 2002 Oct, 66(10), 2209 - 15
Effects of iprodione and fludioxonil on glycerol synthesis and hyphal development in Candida albicans; Ochiai N et al.; We investigated the effects of iprodione and fludioxonil on the pathogenic yeast Candida albicans . Growth of the wild-type IFO1385 strain of C . albicans was inhibited by both fungicides, while Saccharomyces cerevisiae was basically unaffected by them even at a concentration of 25 microg/ml . Both fungicides stimulated glycerol synthesis in C . albicans but not in S . cerevisiae . The antioxidant alpha-tocopherol acetate and the cytochrome P-450 inhibitor piperonyl butoxide antagonized the fungitoxicity of iprodione and fludioxonil in C . albicans . It is known that mutations within the histidine kinase NIK1/OS-1 gene confer resistance to iprodione and fludioxonil in Neurospora crassa, while the fungicide-insensitive S . cerevisiae has only one histidine kinase SLN1 gene in its genome . In contrast, C . albicans has three histidine kinase genes, namely CaSLN1, CaNIK1/COS1, and CaHK1, the null mutants of which were found to impair the hyphal formation . Iprodione and fludioxonil were found to suppress filamentation when the IFO1385 strain was incubated on a solid medium containing fetal bovine serum . These observations suggest that iprodione and fludioxonil interfere with the CaNIK1/COS1 signal transduction pathway, resulting in glycerol synthesis stimulation and the inhibition of hyphal formation.

Biosci Biotechnol Biochem, 2002 Oct, 66(10), 2168 - 75
Cloning and characterization of the nagA gene that encodes beta-n-acetylglucosaminidase from Aspergillus nidulans and its expression in Aspergillus oryzae; Kim S et al.; We isolated a beta-N-acetylglucosaminidase encoding gene and its cDNA from the filamentous fungus Aspergillus nidulans, and designated it nagA . The nagA gene contained no intron and encoded a polypeptide of 603 amino acids with a putative 19-amino acid signal sequence . The deduced amino acid sequence was very similar to the sequence of Candida albicans Hex1 and Trichoderma harzianum Nag1 . Yeast cells containing the nagA cDNA under the control of the GAL1 promoter expressed beta-N-acetylglucosaminidase activity . The chromosomal nagA gene of A . nidulans was disrupted by replacement with the argB marker gene . The disruptant strains expressed low levels of beta-N-acetylglucosaminidase activity and showed poor growth on a medium containing chitobiose as a carbon source . Aspergillus oryzae strain carrying the nagA gene under the control of the improved glaA promoter produced large amounts of beta-N-acetylglucosaminidase in a wheat bran solid culture.

Zh Mikrobiol Epidemiol Immunobiol, 2002 Jul-Aug, (4), 17 - 20
{Phenomenon of selective decrease of colonization (adherence) resistance in the system "Candida albicans-buccal epithelial cells"}; Maianskii AN et al.; In the period of the exacerbation of bronchial asthma an increased adhesiveness (in vitro) of buccal epithelial cells to C . albicans was noted in most children under study (94.7%) . This phenomenon was not observed in children with the exacerbation of gastroduodenitis . The characteristics of natural bacterial colonization of buccal epithelium were equally decreased in both groups of patients . These results are regarded as the consequence of the reactive involvement of the epithelium of mucoid tract in the processes destabilizing homeostasis.

J Med Microbiol, 2002 Nov, 51(11), 929 - 36
Early membrane exposure of phosphatidylserine followed by late necrosis in murine macrophages induced by Candida albicans from an HIV-infected individual; Panagio LA et al.; The hypothesis that Candida albicans isolate (CR1) from an HIV-infected individual induced apoptosis of macrophages was examined by optical microscopy, binding of annexin V-FITC and analyses of DNA degradation (TUNEL tests and agarose gel electrophoresis) . Resident murine peritoneal macrophages co-incubated for 5-15 min with C . albicans CR1 bound annexin V, whereas macrophages incubated with either heat-inactivated strain CR1, C . albicans 577 (isolated from a patient with mucocutaneous candidiasis) or C . albicans FCF14 (a mutant that did not produce proteases and phospholipases) did not bind annexin for up to 2 h of observation . However, macrophages exposed to C . albicans CR1 did not present the pattern of DNA degradation typical of apoptosis . Macrophages became increasingly permeable to propidium iodide from 30 min to 2 h after their exposure to C . albicans CR1 . Most of the phagocytosed C . albicans CR1 yeast cells switched to germ-tubes inside the macrophages after incubation for 1-2 h . These results show that macrophages exposed to C . albicans CR1 presented early signs of apoptosis but progressed to necrosis, and suggest that Candida strains that readily switch to germ-tubes inside those apoptotic cells might have a competitive advantage in vivo because released germ-tubes resist further attack by macrophages.

Z Kardiol, 2002 Sep, 91(9), 715 - 8
{Candida albicans endocarditis with mycotic aneurysms of the lower leg arteries in a patient with mechanical heart valve prosthesis}; Kindel M et al.; We report on a 53-year-old patient in whom the aortic and mitral valves had been replaced . This patient developed Candida albicans sepsis with mycotic aneurysms of the tibial arteries as well as cerebral emboli under immunosuppression . Dispensing with a valve replacement operation, a sustained freedom from recurrence (period of observation currently 24 months) could be attained with antimycotic therapy with amphotericin B and flucytosine i.v . and subsequent long-term therapy with fluconazole p.o . as well as surgical and interventional clearance of the mycotic aneurysms.

Ginecol Obstet Mex, 2002 Sep, 70, 431 - 6
{Study on Candida no-albicans species and its relation to recurrent vulvovaginal candidiasis}; Buitron Garcia R et al.; Genital candidiasis is a frequent pathology among women . It has a 5% incidence rate of presenting itself in a recurrent way, which leads to a longer treatment . Candida gender has various species . The ones, which are the most usual and the cause of vaginal, cervicovaginal, and vulvovaginal candidiasis are: Candida albicans, C . glabrata and C . krusei . Their presence is related to further appearances . A case study was made to identify the species of Candida gender . It was based on the diagnostics made in three Mexico City hospitals on female genital candidiasis cases . The identified and isolated Candida species obtained were: albicans with a 71.91%, C . glabrata with a 11.80%, and C . tropicalis with a 7.87%.

Arch Pharm (Weinheim), 2002 Nov, 335(9), 438 - 42
New 2H-tetrahydro-1, 3, 5-thiadiazine-2-thiones incorporating glycine and glycinamide as potential antifungal agents; Aboul-Fadl T et al.; The new title derivatives (4b-h and 5a-i) were synthesized by reaction of the appropriate primary amine, carbon disulphide, and formaldehyde . These derivatives were prepared in order to study the effects of introducing polar groups at N3 or N5 or at both positions on the biological activity . The compounds were tested for their antifungal activity in vitro against pathogenic (Trichophyton rubrum and Candida albicans), phytopathogenic (Penicillum expansum, Trichoderma hazianum, and Fasarium oxysporum), and aflatoxin-producing (Aspergillus flavus) fungi . These compounds exhibited varied inhibitory effects on growth or sporulation of some tested fungal species.

J Infect Dis, 2002 Dec 15, 186(12), 1815 - 22 Epub 2002 Nov 22.
Delayed clearance of intraabdominal abscesses caused by Candida albicans in tumor necrosis factor-alpha- and lymphotoxin-alpha-deficient mice; Vonk AG et al.; The role of endogenous tumor necrosis factor-alpha (TNF) and lymphotoxin-alpha (LT) in a model of intraabdominal Candida sepsis and abscess formation was investigated . Significantly more abscesses were observed in TNF/LT double knockout (TNF(-/-)LT(-/-)) mice, compared with that in wild-type (TNF(+/+)LT(+/+)) mice . Outgrowth of Candida in abscesses of TNF(-/-)LT(-/-) mice was 10-fold increased on day 14 and 60-fold increased on day 21 of infection . The interleukin-10rcolon;interferon-gamma ratio, measured in supernatants of stimulated splenocytes, shifted from 131 for TNF(-/-)LT(-/-) mice and 13.9 for TNF(+/+)LT(+/+) mice on day 8 to 0.11 for TNF(-/-)LT(-/-) mice and 11.66 for TNF(+/+)LT(+/+) mice on day 14 of infection . The diminished host resistance is explained by an impaired extracellular killing capacity of granulocytes and a delayed development of a T helper 1 response in TNF(-/-)LT(-/-) mice . In conclusion, TNF and LT are critical to the stimulation of effector cells that leads to elimination of Candida from abscesses.

Planta, 2002 Dec, 216(2), 227 - 34 Epub 2002 Aug 21.
Enzymatic specificity of three ribosome-inactivating proteins against fungal ribosomes, and correlation with antifungal activity; Park SW et al.; Ribosome-inactivating proteins (RIPs) are enzymes that cleave a specific adenine base from the highly conserved sarcin/ricin (S/R) loop of the large ribosomal RNA, thus arresting protein synthesis at the translocation step . In the present study, we employed three RIPs to dissect the antifungal activity of RIPs as plant defense proteins . We measured the catalytic activity of RAT (the catalytic A-chain of ricin from Ricinus communis L.), saporin-S6 (from Saponaria officinalis L.), and ME (RIP from Mirabilis expansa R&P) against intact ribosomal substrates isolated from various pathogenic fungi . We further determined the enzymatic specificity of these three RIPs against fungal ribosomes, from Rhizoctonia solani Kuhn, Alternaria solani Sorauer, Trichoderma reesei Simmons and Candida albicans Berkhout, and correlated the data with antifungal activity . RAT showed the strongest toxicity against all tested fungal ribosomes, except for the ribosomes isolated from C . albicans, which were most susceptible to saporin . RAT and saporin showed higher enzymatic activity than ME against ribosomes from all of the fungal species assayed, but did not show detectable antifungal activity . In contrast, ME showed substantial inhibitory activity against fungal growth . Using N-hydroxysuccinimide-fluorescein labeling of RIPs and fluorescence microscopy, we determined that ME was targeted to the surface of fungal cells and transferred into the cells . Thus, ME caused ribosome depurination and subsequent fungal mortality . In contrast, saporin did not interact with fungal cells, correlating with its lack of antifungal activity.

Mol Cell Biol, 2002 Dec, 22(24), 8669 - 80
Ash1 protein, an asymmetrically localized transcriptional regulator, controls filamentous growth and virulence of Candida albicans; Inglis DO et al.; In response to a number of distinct environmental conditions, the fungal pathogen Candida albicans undergoes a morphological transition from a round, yeast form to a series of elongated, filamentous forms . This transition is believed to be critical for virulence in a mouse model of disseminated candidiasis . Here we describe the characterization of C . albicans ASH1, a gene that encodes an asymmetrically localized transcriptional regulatory protein involved in this response . We show that C . albicans ash1 mutants are defective in responding to some filament-inducing conditions . We also show that Ash1p is preferentially localized to daughter cell nuclei in the budding-yeast form of C . albicans cell growth and to the hyphal tip cells in growing filaments . Thus, Ash1p "marks" newly formed cells and presumably directs a specialized transcriptional program in these cells . Finally, we show that ASH1 is required for full virulence of C . albicans in a mouse model of disseminated candidiasis.

Ann Biol Clin (Paris), 2002 Nov-Dec, 60(6), 711 - 4
{Detection of circulating Candida albicans mannan and antimannan antibodies: useful for diagnosis of deep seated candidiasis}; Eloy O et al.; In deep seated candidiasis, only 40% of blood cultures are positive . The aim of the study was to investigate circulating Candida albicans mannan and anti-mannan antibodies as a possible help for the diagnosis of deep seated candidiasis . We have compared the results to the detection of IgM by Elisa and antibodies by immunoflourescence . The best tests, in accord to their sensitivity and specificity, are the mannan antigenemia (43% and 100%) and IgM (86% and 100%) and have to be used together.

Curr Biol, 2002 Nov 19, 12(22), R782 - 4
Fungal mating: Candida albicans flips a switch to get in the mood; Hull CM et al.; The fungal pathogen Candida albicans can mate under highly controlled conditions . It can also undergo phenotypic switching . A recent discovery joins these disparate processes to reveal that 'opaque' switch variants mate 10(6) times better than 'white' variants.

Exp Dermatol, 2002, 11 Suppl 1, 13 - 6
Mechanisms underlying UV-induced immune suppression: implications for sunscreen design; Ullrich SE et al.; The ultraviolet (UV) radiation present in sunlight is immune-suppressive . Recently we showed that solar-simulated UV radiation (UVA + UVB; 295-400 nm), applied after immunization, suppressed immunological memory and the elicitation of delayed-type hypersensitivity to the common opportunistic pathogen, Candida albicans . Further, we found that wavelengths in the UVA region of the solar spectrum (320-400 nm), devoid of UVB, were equally effective in activating immune suppression as UVA + UVB radiation . Here we report on the mechanisms involved . No immune suppression was found in UV-irradiated mice injected with monoclonal anti-interleukin (IL)-10 antibody, or mice exposed to solar-simulated UV radiation and injected with recombinant IL-12 . Antigen-specific suppressor T cells were found in the spleens of mice exposed to UVA + UVB radiation . Applying liposomes containing bacteriophage T4N5 to the skin of mice exposed to solar-simulated UVA + UVB radiation or mice exposed to UVA radiation blocked immune suppression, demonstrating an essential role for UV-induced DNA damage in the suppression of established immune reactions . These findings indicate that UV radiation activates similar immunological pathways to suppress the induction, or the elicitation, of the immune response.

J Nat Prod, 2002 Nov, 65(11), 1660 - 3
Pharacine, a natural p-cyclophane and other indole derivatives from Cytophaga sp . strain AM13.1; Shaaban M et al.; From the ethyl acetate extract of the bacterial strain Cytophaga sp . AM13.1, among many known compounds, the new natural products 2,5-bis(3-indolylmethyl)pyrazine (2) and a highly symmetrical p-cyclophane named pharacine (5) were identified . In addition, tryptamine isovalerate (1) and p-hydroxyphenylacetamide (4), known as plant metabolites, were isolated and characterized from a microorganism for the first time . The new natural products showed no activity against three microalgae, the fungus Mucor miehei, the yeast Candida albicans, and the bacteria Staphylococcus aureus, Bacillussubtilis, Escherichia coli, and Streptomyces viridochromogenes.

J Nat Prod, 2002 Nov, 65(11), 1598 - 604
Online analysis of xestodecalactones A-C, novel bioactive metabolites from the fungus Penicillium cf . montanense and their subsequent isolation from the sponge Xestospongia exigua; Edrada RA et al.; Fungal isolates of Penicillium cf . montanense were obtained from the marine sponge Xestospongia exiguacollected from the Bali Sea, Indonesia . Culture filtrates of the fungi yielded three novel decalactone metabolites, xestodecalactones A, B, and C (1, 2a, and 2b), consisting of 10-membered macrolides with a fused 1,3-dihydroxybenzene ring . Online HPLC-NMR, ESI-MS/MS, and -CD spectra were acquired, and the structures of the new compounds were established and confirmed on the basis of offline NMR spectroscopic ((1)H, (13)C, COSY, ROESY, (1)H-detected direct and long-range (13)C-(1)H correlations) and mass spectrometric (EIMS) data . Quantum chemical calculations of the CD spectra proved to be difficult because of the conformational flexibility of the xestodecalactones . These compounds, of which 2a and 2b, due to the additional stereocenter at C-9, are diastereomeric compounds, are structurally related to a number of biologically active metabolites found in terrestrial fungal strains . Compound 2a was found to be active against the yeast Candida albicans.

J Immunol, 2002 Dec 1, 169(11), 6298 - 308
B7/CD28-dependent CD4+CD25+ regulatory T cells are essential components of the memory-protective immunity to Candida albicans; Montagnoli C et al.; Protective immunity to the fungus Candida albicans is mediated by Ag-specific Th1 cells . Paradoxically, some Th2 cytokines are required for the maintenance of Th1-mediated immune resistance to the fungus . Therefore, in addition to the Th1/Th2 balance, other mechanisms seem to be involved in the regulation of Th1 immunity to the fungus . Here we show that CD4(+)CD25(+) T cells, negatively regulating antifungal Th1 reactivity, are generated in mice with candidiasis . CD4(+)CD25(+) T cells were not generated in B7-2- or CD28-deficient mice or in condition of IL-10 signaling deficiency . Accordingly, although capable of efficiently restricting the fungal growth, these mice experienced inflammatory pathology and were incapable of resistance to reinfection . CD4(+)CD25(+) T cells poorly proliferated in vitro; were highly enriched for cells producing IL-4, IL-10, and TGF-beta; and required IL-10-producing, Candida hypha-activated dendritic cells for generation . Adoptive transfer of CD4(+)CD25(+) T cells or IL-10-producing dendritic cells restored resistance to reinfection and decreased inflammation in B7-2-deficient mice . These results show that oral tolerance induced by Candida hyphae is required for the occurrence of long-lasting protective immunity after yeast priming . The implication is that preventing reactivation rather than favoring sterilizing immunity to ubiquitous fungal pathogens may represent the ultimate expectation of vaccine-based strategies.

Medicine (Baltimore), 2002 Nov, 81(6), 425 - 33
Secular trends of candidemia over 12 years in adult patients at a tertiary care hospital; Garbino J et al.; The incidence of fungal infections has been increasing for the last 3 decades, especially among neutropenic, cancer, and critically ill patients . These infections are associated with high mortality rates . We retrospectively reviewed medical charts of adult patients with fungemia from 1989 to 2000 at our institution . The characteristics of the population groups served by the hospital were described . Of 328 patients with fungemia, we reviewed 315 (96%) medical records, and focused on those with candidemia (n = 294) . The species distribution in patients with candidemia showed that the most commonly identified species were Candida albicans (66%), followed by C . glabrata (17%), and C . parapsilosis (6%) . The incidence of candidemia ranged from 0.2 to 0.46 per 10,000 patient-days with the highest incidence in 1993 and the lowest in 1997 . Although most studies show an increased incidence of candidemia, we observed a reduction over the study period . Furthermore, we observed no shift from C . albicans to non-albicans Candida species despite a significant increase in the use of fluconazole . The overall mortality among patients with candidemia was 44%, with the highest rate in patients over 65 years (52%) . Factors independently associated with higher mortality were patient age greater than 65 years, intensive care unit admission, and underlying cancer.

Clin Infect Dis, 2002 Dec 1, 35(11), 1316 - 20 Epub 2002 Nov 07.
Candida albicans sternal wound infections: a chronic and recurrent complication of median sternotomy; Malani PN et al.; Eleven patients developed deep sternal wound infections due to Candida albicans after undergoing coronary artery bypass grafting (CABG) and were assessed . Six had sternal osteomyelitis, 1 had osteomyelitis and mediastinitis, and 4 had deep wound infections that probably involved bone . Seven patients experienced onset of infection within 28 days of CABG, but 4 experienced onset 48-150 days after CABG . Infections were characterized by a chronic, indolent course requiring prolonged treatment with an antifungal agent . Delay in initiating antifungal therapy was common . All patients were treated with fluconazole, and 1 also received amphotericin B . Six patients underwent incision and drainage, with or without wire removal, and 3 underwent sternectomy with placement of a muscle flap . Of 10 patients for whom follow-up data were available, 7 were cured after initial therapy (median duration of treatment, 6 months), and 3 experienced a relapse and required a second course of fluconazole.

Infect Immun, 2002 Dec, 70(12), 7081 - 8
Vaginal and oral epithelial cell anti-Candida activity; Nomanbhoy F et al.; Candida albicans is the causative agent of acute and recurrent vulvovaginal candidiasis (VVC), a common mucosal infection affecting significant numbers of women in their reproductive years . While any murine host protective role for cell-mediated immunity (CMI), humoral immunity, and innate resistance by neutrophils against the vaginal infection appear negligible, significant in vitro growth inhibition of Candida species by vaginal and oral epithelial cell-enriched cells has been observed . Both oral and vaginal epithelial cell anti-Candida activity has a strict requirement for cell contact to C . albicans with no role for soluble factors, and oral epithelial cells inhibit C . albicans through a cell surface carbohydrate moiety . The present study further evaluated the inhibitory mechanisms by murine vaginal epithelial cells and the fate of C . albicans by oral and vaginal epithelial cells . Similar to human oral cells, anti-Candida activity produced by murine vaginal epithelial cells is unaffected by enzymatic cleavage of cell surface proteins and lipids but sensitive to periodic acid cleavage of surface carbohydrates . Analysis of specific membrane carbohydrate moieties, however, showed no role for sulfated polysaccharides, sialic acid residues, or glucose and mannose-containing carbohydrates, also similar to oral cells . Staining for live and dead Candida in the coculture with fluorescein diacetate (FDA) and propidium iodide (PI), respectively, showed a clear predominance of live organisms, suggesting a static rather than cidal action . Together, the results suggest that oral and vaginal epithelial cells retard or arrest the growth rather than kill C . albicans through an as-yet-unidentified carbohydrate moiety in a noninflammatory manner.

Infect Immun, 2002 Dec, 70(12), 7073 - 80
Interleukin-18 and gamma interferon production by oral epithelial cells in response to exposure to Candida albicans or lipopolysaccharide stimulation; Rouabhia M et al.; Oral candidiasis is a collective name for a group of disorders caused by the dimorphic fungus Candida albicans . Host defenses against C . albicans essentially fall into two categories: specific immune mechanisms and local oral mucosal epithelial cell defenses . Since oral epithelial cells secrete a variety of cytokines and chemokines in response to oral microorganisms and since C . albicans is closely associated with oral epithelial cells as a commensal organism, we wanted to determine whether interleukin-18 (IL-18) and gamma interferon (IFN-gamma) were produced by oral epithelial cells in response to C . albicans infection and lipopolysaccharide (LPS) stimulation . Our results showed that IL-18 mRNA and protein were constitutively expressed by oral epithelial cells and were down-regulated by Candida infections but increased following LPS stimulation . Both C . albicans and LPS significantly decreased pro-IL-18 (24 kDa) levels and increased active IL-18 (18 kDa) levels . This effect was IL-1beta-converting-enzyme dependent . The increase in active IL-18 protein levels promoted the production of IFN-gamma by infected cells . No effect was obtained with LPS . Although produced only at an early stage, secreted IFN-gamma seemed to be a preferential response by oral epithelial cells to C . albicans growth . These results provide additional evidence for the contribution of oral epithelial cells to local (direct contact) and systemic (IL-18 and IFN-gamma production) defense against exogenous stimulation such as C . albicans infection or LPS stimulation.

Mol Genet Genomics, 2002 Nov, 268(3), 418 - 24 Epub 2002 Oct 19.
Site-specific targeting of exogenous DNA into the genome of Candida albicans using the FLP recombinase; Sanchez-Martinez C et al.; We have created a system that utilizes the FLP recombinase of Saccharomyces cerevisiae to reversibly introduce exogenous cloned DNA at defined locations into the Candida albicans genome . Recombination target (FRT) sites and the FLP gene can be introduced permanently at defined locations using homologous recombination . FLP recombinase is provided as needed through the regulated expression of its gene using the MAL2 promoter . Exogenous DNA is introduced on a cloning vector that is unable to replicate in C . albicans, and contains an FRT site and a selectable marker (URA3) . Transformation by the lithium acetate or electroporation procedure is sufficient to obtain site-specific integration . This system permits rapid and precise excision of the introduced DNA when needed . It should facilitate studies on C . albicans genome structure and function, simplifying a wide range of chromosomal cloning applications, and generally enhancing the utility of C . albicans as a model organism for the study of fungal pathogenicity.

Antimicrob Agents Chemother, 2002 Dec, 46(12), 4016 - 8
New automated method for determining postantifungal effect of amphotericin B against Candida species: effects of concentration, exposure time, and area under the curve; Chryssanthou E et al.; The postantifungal effect (PAFE) of amphotericin B was determined with a BacT/Alert automated system, which is based on the colorimetric detection of CO(2) . The levels of accuracy and precision of the automated method were high . Longer PAFEs were obtained for Candida albicans (P < 0.001), C . glabrata (P < 0.01), and C . krusei (P < 0.01) at increasing amphotericin B concentrations and exposure times.

Antimicrob Agents Chemother, 2002 Dec, 46(12), 3869 - 76
Synthetic analogues of beta-1,2 oligomannosides prevent intestinal colonization by the pathogenic yeast Candida albicans; Dromer F et al.; The pathogenic yeast Candida albicans displays at its cell surface beta-1,2 oligomannosides (beta-1,2-Mans) . In contrast to the ubiquitous alpha-Mans, beta-1,2-Mans bind to galectin-3, a major endogenous lectin expressed on epithelial cells . The specific role of beta-1,2-Mans in colonization of the gut by C . albicans was assessed in a mouse model . A selected virulent strain of C . albicans (expressing more beta-1,2-Man epitopes) induced more intense and sustained colonization than an avirulent strain (expressing less beta-1,2-Man epitopes) . Synthetic (Sigma) beta-and alpha-linked tetramannosides with antigenicities that mimicked the antigenicities of C . albicans-derived oligomannosides were then constructed . Oral administration of Sigmabeta-1,2-Man (30 mg/kg of body weight) prior to inoculation with the virulent strain resulted in almost complete eradication of yeasts from stool samples, whereas administration of Sigmaalpha-Man at the same dose did not . As most cases of human systemic candidiasis are endogenous in origin, this first demonstration that a synthetic analogue of a yeast adhesin can prevent yeast colonization in the gut opens the possibility of new prophylactic strategies.

Antimicrob Agents Chemother, 2002 Dec, 46(12), 3695 - 705
Drug susceptibilities of yeast cells are affected by membrane lipid composition; Mukhopadhyay K et al.; In the present study we have exploited isogenic erg mutants of Saccharomyces cerevisiae to examine the contribution of an altered lipid environment on drug susceptibilities of yeast cells . It is observed that erg mutants, which possess high levels of membrane fluidity, were hypersensitive to the drugs tested, i.e., cycloheximide (CYH), o-phenanthroline, sulfomethuron methyl, 4-nitroquinoline oxide, and methotrexate . Most of the erg mutants except mutant erg4 were, however, resistant to fluconazole (FLC) . By using the fluorophore rhodamine-6G and radiolabeled FLC to monitor the passive diffusion, it was observed that erg mutant cells elicited enhanced diffusion . The addition of a membrane fluidizer, benzyl alcohol (BA), to S . cerevisiae wild-type cells led to enhanced membrane fluidity . However, a 10 to 12% increase in BA-induced membrane fluidity did not alter the drug susceptibilities of the S . cerevisiae wild-type cells . The enhanced diffusion observed in erg mutants did not seem to be solely responsible for the observed hypersensitivity of erg mutants . In order to ascertain the functioning of drug extrusion pumps encoding the genes CDR1 (ATP-binding cassette family) and CaMDR1 (MFS family) of Candida albicans in a different lipid environment, they were independently expressed in an S . cerevisiae erg mutant background . While the fold change in drug resistance mediated by CaMDR1 remained the same or increased in erg mutants, susceptibility to FLC and CYH mediated by CDR1 was increased (decrease in fold resistance) . Our results demonstrate that between the two drug extrusion pumps, Cdr1p appeared to be more adversely affected by the fluctuations in the membrane lipid environment (particularly to ergosterol) . By using 6-{(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino-hexanoyl} sphingosyl phosphocholine (a fluorescent analogue of sphingomyelin), a close interaction between membrane ergosterol and sphingomyelin which appears to be disrupted in erg mutants is demonstrated . Taken together it appears that multidrug resistance in yeast is closely linked to the status of membrane lipids, wherein the overall drug susceptibility phenotype of a cell appears to be an interplay among drug diffusion, extrusion pumps, and the membrane lipid environment.

J Inorg Biochem, 2002 Nov 25, 92(3-4), 141 - 8
Biological activity of palladium(II) and platinum(II) complexes of the acetone Schiff bases of S-methyl- and S-benzyldithiocarbazate and the X-ray crystal structure of the {Pd(asme)2} (asme=anionic form of the acetone Schiff base of S-methyldithiocarbazate) complex; Akbar Ali M et al.; Palladium(II) and platinum(II) complexes of general empirical formula, {M(NS)(2)} (NS=uninegatively charged acetone Schiff bases of S-methyl- and S-benzyldithiocarbazate; M=Pt(II) and Pd(II)) have been prepared and characterized by a variety of physicochemical techniques . Based on conductance, IR and electronic spectral evidence, a square-planar structure is assigned to these complexes . The crystal and molecular structure of the {Pd(asme)(2)} complex (asme=anionic form of the acetone Schiff base of S-methyldithiocarbazate) has been determined by X-ray diffraction . The complex has a distorted cis-square planar structure with the ligands coordinated to the palladium(II) ions as uninegatively charged bidentate NS chelating agents via the azomethine nitrogen and the mercaptide sulfur atoms . The distortion from a regular square-planar geometry is attributed to the restricted bite angles of the ligands . Antimicrobial tests indicate that the Schiff bases exhibit strong activities against the pathogenic bacteria, Bacillus subtilis (mutant defective DNA repair), methicillin-resistant Staphylococcus aureus, B . subtilis (wild type) and Pseudomonas aeruginosa and the fungi, Candida albicans (CA), Candida lypotica (2075), Saccharomyces cerevisiae (20341) and Aspergillus ochraceous (398)-the activities exhibited by these compounds being greater than that of the standard antibacterial and antifungal drugs, streptomycin and nystatin, respectively . The palladium(II) and platinum(II) complexes are inactive against most of these organisms but, the microbe, Pseudomonas aeruginosa shows strong sensitivity to the platinum(II) complexes . Screening of the compounds for their cytotoxicities against T-lymphoblastic leukemia cancer cells has shown that the acetone Schiff base of S-methyldithiocarbazate (Hasme) exhibits a very weak activity, whereas the S-benzyl derivative (Hasbz) is inactive . However, the palladium(II) complexes exhibit strong cytotoxicities against this cancer; their activities being more than that of the standard anticancer drug, tamoxifen . The {Pt(asme)(2)} complex exhibits a very weak cytotoxicity, whereas {Pt(asbz)(2)} is inactive against leukemic cells.

Curr Infect Dis Rep, 2002 Dec, 4(6), 514 - 519
Pathogenesis of Recurrent Vulvovaginal Candidiasis; Sobel JD; Recurrent vulvovaginal candidiasis (RVVC) is by no means uncommon and is a source of considerable physical suffering, in addition to serving as a major therapeutic challenge . The syndrome is multifactorial in etiology, hence management strategies must recognize the complex etiologic pathways . Considerable progress has been made in identifying secondary causes, including biologic and host factors . Specifically, Candida microbiologic studies have revealed that azole resistance in Candida albicans is rare and infection by less sensitive non-albicans Candida species is uncommon . At least half the women with RVVC have no identifiable host or microbial predisposing factors, and an immune-based hypothesis has been generated.

Microbiology, 2002 Nov, 148(Pt 11), 3743 - 54
Differential secretion of Sap4-6 proteins in Candida albicans during hyphae formation; Chen YC et al.; Secreted aspartyl proteinases (Saps) from Candida albicans are encoded by a multi-gene family and are considered to be putative virulence factors for candidiasis . SAP4-6 mRNAs were first detected during hyphae formation and were assumed to play roles in the development of disseminated candidiasis . Recombinant Sap proteins (Sap2-6) were prepared and specific antibodies were generated against Sap2-6 . The presence of Sap4, Sap5 and Sap6, but not Sap2 or Sap3, was demonstrated in culture supernatants of C . albicans after induction of hyphae formation . In parallel to hyphae formation, Sap5 (approximately 40 kDa) was detected as early as approximately 6 h after induction at neutral pH, and Sap4/6 (approximately 43 kDa) were detected after approximately 24 h when the culture medium became acidic . The differential secretion of Sap5 and Sap4/6 was affected when the culture medium pH was buffered at pH 6.5 or pH 4.5 . In addition, intracellular pools of Sap4-6 seem to exist, and protein is not necessary for Sap4-6 induction . This study provides the first evidence that Sap4-6 proteins in C . albicans are differentially produced and secreted during hyphae formation.

Microbiology, 2002 Nov, 148(Pt 11), 3705 - 13
Copper- and zinc-containing superoxide dismutase (Cu/ZnSOD) is required for the protection of Candida albicans against oxidative stresses and the expression of its full virulence; Hwang CS et al.; Copper- and zinc-containing superoxide dismutase (Cu/ZnSOD) is suspected to be one of the anti-oxidant enzymes and virulence determinants active in some pathogenic micro-organisms . To elucidate the role of Cu/ZnSOD in the major human fungal pathogen Candida albicans, its gene, designated SOD1, was disrupted by the URA-blaster technique . The resulting sod1/sod1 mutant showed delayed hyphal growth on Spider medium but could still form hyphae on other solid media or in liquid media, particularly in response to serum . Moreover, the sod1/sod1 mutant was more sensitive to menadione, a redox-cycling agent, than the isogenic wild-type strain, although it still showed an adaptive oxidative stress response . Furthermore, the sod1/sod1 mutant cells exhibited slow growth in minimal medium when compared to the wild-type cells, but their growth was restored by the addition of lysine to the medium . Interestingly, C . albicans cells lacking Cu/ZnSOD showed increased susceptibility to macrophage attack and had attenuated virulence in mice . Thus, these results suggest that Cu/ZnSOD is required for the protection of C . albicans against oxidative stresses and for the full virulence of the organism to be expressed.

J Prosthet Dent, 2002 Sep, 88(3), 263 - 7
Susceptibility of Candida albicans isolates from the oral cavities of HIV-positive patients to histatin-5; Nikawa H et al.; STATEMENT OF PROBLEM: Oral surfaces, including the denture-fitting surface, may serve as a reservoir for disseminated candidal infections, particularly in immunocompromised hosts such as patients with AIDS . Histatins are a group of small, cationic antifungal peptides present in human saliva . There is limited information on the antifungal activity of peptides against Candida albicans isolates from HIV-positive patients . PURPOSE: This study investigated the fungicidal effects of histatin-5 against oral isolates of C . albicans from HIV-positive and HIV-negative patients . MATERIAL AND METHODS: An isolate of C . albicans from each of 2 HIV-positive patients (both male) and 3 HIV-negative patients (2 male and 1 female) was obtained . American Type Culture Collection 90028 served as a reference strain . All isolates were identified with sugar assimilation tests and the germ tube test . Fungicidal assays were performed on exponential C . albicans cells in the presence or absence of 0.315 to 50 microm of histatin-5 . Numerical data were subjected to 1-way analysis of variance and Tukey's multiple range test (P<.05) . RESULTS: Histatin-5 (50 microm) killed more than 95% of C . albicans isolates from HIV-negative patients and more than 90% of isolates from the reference strain . The same treatment induced 75.3% and 66.1% loss of viability in C . albicans isolates taken from HIV-positive patients (A1 and A2 cells, respectively) . The difference between the fungicidal effects in the HIV-positive and HIV-negative groups was significant . (P<.05) . CONCLUSION: Within the limited population of this study, C . albicans isolates from the oral cavities of HIV-positive patients were less sensitive to histatin-5 than oral isolates from HIV-negative patients.

What Is Molecular Biology?, What Is Activated Sludge?, What Is Cell Biology?, What Is Listeria Monocytogenes?, What Is Antibiotic?, c, Bacteriology, n, Bacteria, e, Bacterium, e, Microbiology, o, Microorganism, i, Pichia, o, Streptococcal, s, Biodegradation, i, Ciprofloxacin, a, Bacteriological, s, Erythromycin, e, Erythromycin, c, Microbiological, a, Bacillus subtilis, o, Meningococcus, i, Bacteria, c, Escherichia coli, c, Bacteriological, n, Microorganisms, s, Escherichia coli, r, S. cerevisiae, o, Haemophilus, a, Cell suspensions, r, Bacillus, r, Microbiological, s, Antibiotic treatment

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