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 deficiencie