Mycoses, 2000 Oct, 43(9-10), 349 - 53 The frequency of Candida parapsilosis in onychomycosis . An epidemiological survey in Israel; Segal R et al.; Candida albicans is regarded as the major pathogen in yeast-induced onychomycosis . Based on our impression of an increasing prevalence of Candida parapsilosis in this disease, we examined the data of two mycology laboratories in the same geographic location, from 1994 to 1996 in one (centre A) and for 1995 (6 months) in the other (centre B) . A total of 954 and 230 toenails and 621 and 190 fingernails, respectively, underwent KOH microscopy and culture studies in each centre . Positive findings were noted in 45 and 65% of the toenails and 44 and 72% of the fingernails, respectively . In the toenails, Candida spp . were found in 22 and 15%, respectively, and in the fingernails, in 77 and 63%, respectively . The most frequent Candida species was C . parapsilosis (39.5% in toenails, 36.7% in fingernails), followed by C . albicans (19.5% in toenails, and 34.4% in fingernails) . These results demonstrate a higher frequency of isolation of C . parapsilosis compared with C . albicans in onychomycosis . This might have important therapeutic implications. Ned Tijdschr Geneeskd . 2000 Oct 28;144(44):2103. {Diagnostic image (9) . Chronic mucocutaneous candidiasis}; van der Meer JW et al.; An 18-year-old man had lifelong mucocutaneous lesions from which Candida albicans was persistently cultured . Upon fluconazole treatment the lesions diminished notably. J Antimicrob Chemother, 2000 Dec, 46(6), 1001 - 8 Impact of fluconazole prophylaxis on fungal colonization and infection rates in neutropenic patients . The Canadian Fluconazole Study; Laverdiere M et al.; Fungal colonization profiles from four different anatomical sites were evaluated in 266 neutropenic cancer patients receiving intensive cytotoxic therapy for acute leukaemia or for autologous marrow transplantation . At the beginning of chemotherapy patients were allocated randomly to receive oral fluconazole 400 mg daily or an identical placebo until prophylaxis failure or marrow recovery . Candida albicans colonization was reduced from 30 to 10% in the fluconazole recipients while it increased from 32 to 57% in the placebo patients (P<0.001) . By the end of prophylaxis, colonization with non-albicans Candida species increased from 7 to 21% and 8 to 18% in the fluconazole and placebo patients, respectively (P = 0.396) . Although Candida glabrata was isolated more frequently at the end of the prophylactic period in the fluconazole patients than in the placebo patients (16 versus 7%), only one definite invasive C . glabrata infection was noted . Overall, definite invasive fungal infections were documented in 26 patients {four fluconazole versus 22 placebo patients (P< or =0.001)} . In 23 (92%) patients the infections were caused by persistently colonizing or newly acquired organisms . While probable invasive fungal infections were noted in five fluconazole patients, 10 placebo patients were also affected (P = 0.19) . An end-of-prophylaxis colonization index >0.25 was 76% sensitive but only 69% specific for invasive fungal infection . However, a colonization index < or =0.25 at baseline had a negative predictive value of 88% for development of invasive fungal infection . Fluconazole prophylaxis decreased colonization by fungi and subsequent invasive fungal infections in neutropenic cancer patients. J Clin Microbiol, 2000 Dec, 38(12), 4626 - 8 Tween 80 opacity test responses of various Candida species; Slifkin M; A total of 111 Candida isolates representing 11 species were examined for their respective responses to a Tween 80 opacity test . The strains of Candida albicans and C . tropicalis that were examined produced an opacity response around their colonies at 2 to 3 days postinoculation . A second group of Candida species yielded a halo around their colonies at 8 to 10 days postinoculation . The remaining Candida species did not produce a positive test response through 10 days postinoculation . The strains of C . dubliniensis were easily differentiated from strains of C . albicans by this test . The Tween 80 opacity test is simple and economical to prepare and is easy to interpret. J Clin Microbiol, 2000 Dec, 38(12), 4554 - 9 Comparative genotyping of Candida albicans bloodstream and nonbloodstream isolates at a polymorphic microsatellite locus; Dalle F et al.; Molecular typing studies have shown that the predominant form of reproduction of Candida albicans is clonal and that, in a majority of situations, persistent or recurrent infections are due to a unique strain . Characterization of distinct subpopulations and correlation with clinical features may thus be important to understanding the pathogenesis of candidiasis . In a clonal model, a unique polymorphic marker may identify populations with different biological properties . We therefore compared 48 bloodstream isolates and 48 nonbloodstream matched strains of C . albicans at the elongation factor 3-encoding gene (CEF3) polymorphic microsatellite locus of C . albicans . Sizing of the alleles was performed by automated capillary electrophoresis . A new, 137-bp allele was characterized, and seven nondescribed combinations were observed, resulting in 15 and 11 distinct CEF3 profiles in bloodstream and control strains, respectively . Genotypes 126-135, 130-136, and 131-131 accounted for 60.4% of both bloodstream and control strains . Four bloodstream isolates but no control strains displayed the 135-135 combination . None of the other genotypes was present at an increased frequency in bloodstream isolates . Bloodstream and nonbloodstream strains of C . albicans thus have a heterogeneous structure at the CEF3 locus, with three major and multiple minor allelic combinations. J Clin Microbiol, 2000 Dec, 38(12), 4503 - 10 Identification and phylogenetic relationship of the most common pathogenic Candida species inferred from mitochondrial cytochrome b gene sequences; Yokoyama K et al.; We sequenced a 396-bp region of the mitochondrial cytochrome b gene of the most common clinically important Candida species: Candida albicans, C . glabrata, C . parapsilosis, C . tropicalis, C . krusei, and C . lusitaniae . The recently described species of Candida, C . dubliniensis, associated with mucosal candidiasis in human immunodeficiency virus-infected individuals, was also included . Two to five strains of each species were examined . Some species represented intraspecies variation, which was not more than 1.8% (DNA) . However, interspecies variations were more than 10 and 7%, respectively, for DNA and amino acid sequences . Multiple alignments of nucleotide and deduced amino acid sequences revealed species-specific nucleotides and amino acids . Nucleotide- and amino acid-based phylogenetic trees were constructed and are discussed . Using the database, it is possible to identify presumptive Candida species within a working day. J Antibiot (Tokyo), 2000 Sep, 53(9), 920 - 7 FR901469, a novel antifungal antibiotic from an unidentified fungus No.11243 . II . In vitro and in vivo activities; Fujie A et al.; FR901469 is a water-soluble macrocyclic lipopeptidolactone (C71H116N14O23) that has inhibitory activity against 1,3-beta-glucan synthase and exhibits in vitro and in vivo antifungal activity against both Candida albicans and Aspergillus fumigatus . The MICs of FR901469 against Candida albicans FP633 and Aspergillus fumigatus FP1305 in a micro-broth dilution test were 0.63 and 0.16 microg/ml, respectively . FR901469 showed excellent efficacy by subcutaneous injection against both Candida albicans and Aspergillus fumigatus in a murine systemic infection mode, with ED50s of 0.32 and 0.2 mg/kg, respectively . This compound also showed potent anti-Pneumocystis activity in the nude mice model with experimental Pneumocystis pneumonia . The hemolytic activity of FR901469 towards mouse red blood cells, is about 30-fold weaker than that of amphotericin B. J Antibiot (Tokyo), 2000 Sep, 53(9), 912 - 9 FR901469, a novel antifungal antibiotic from an unidentified fungus No.11243 . I . Taxonomy, fermentation, isolation, physico-chemical properties and biological properties; Fujie A et al.; FR901469 is a novel antifungal antibiotic produced by an unidentified fungus No.11243 . This compound was isolated from the culture broth by solvent extraction, HP-20 and YMC ODS gel column chromatography, and lyophilization . FR901469 is a white powder which melts at 182 approximately 187 degrees C and possesses the molecular formula C71H116N14O23 . This compound has good water solubility . FR901469 inhibited the activity of 1,3-beta-glucan synthase from Candida albicans with an IC50 value of 0.05 microg/ml, and displayed greater inhibitory activity than other 1,3-beta-glucan synthase inhibitors such as, WF11899A, echinocandin B, aculeacin A, and papulacandin B. Z Naturforsch {C}, 2000 Sep-Oct, 55(9-10), 785 - 9 Chemical composition and biological activity of propolis from Brazilian meliponinae; Velikova M et al.; Twenty-one propolis samples produced by 12 different Meliponinae species were analyzed by GC-MS . Several chemical types of stingless bees' propolis could be grouped, according to the prevailing type of compounds like: 'gallic acid", "diterpenic" and "triterpenic" types . The results confirm that neither the bee species nor the geographical location determine the chemical composition of Meliponinae propolis and the choice of its plant source, respectively . This could be explained by the fact that Meliponinae forage over short distances (maximum 500 m) and thus use as propolis source the first plant exudate they encounter during their flights . The antibacterial, antifungal and cytotoxic activities of the samples were also investigated . Most samples had weak or no activity against E . coli, weak action against Candida albicans . Some of them showed significant activity against St . aureus., presumably connected to the high concentration of diterpenic acids . Samples rich in diterpenic acids possessed also high cytotoxic activity (Artemia salina test). Mycoses, 2000, 43 Suppl 1, 29 - 35 {Asteroid bodies: host-pathogen reactions in mycoses}; Muller J; A review of the literature on asteroid bodies (Splendore-Hoeppli phenomenon) as well as an immunoelectronmicroscopic study on asteroid bodies with Candida albicans and electronmicroscopic observations on asteroid bodies with Aspergillus are presented . The following definitions are proposed: Asteroid bodies with Candida albicans: precipitate consisting of fungal antigen and antibodies of host origin in light microscopically visible dimension deposited on the cell wall surface of Candida cells in parasitic condition . Asteroid bodies with Aspergillus: precipitate consisting of fungal antigen, antibodies of host origin and necrotic cellular detritus originated from cells of host defense, in light microscopically visible dimension, deposited on the cell wall surface of abortive Aspergillus cells, reduced in longitudinal growth, in parasitic condition. J Pept Res, 2000 Nov, 56(5), 275 - 82 Divergent solid-phase synthesis and candidacidal activity of MUC7 D1, a 51-residue histidine-rich N-terminal domain of human salivary mucin MUC7; Satyanarayana J et al.; Domain 1 of the low-molecular-weight human salivary mucin, designated MUC7 D1, spans the 51 N-terminal amino acid residues . This domain contains a 15-residue basic histidine-rich subdomain (R3-Q17) which has 53% sequence similarity to histatin 5 (Hsn-5), a salivary molecule known to exert potent in vitro cidal activity against Candida albicans and many other medically important fungi . The MUC7 D1-15mer and its derivatives have previously been synthesized in our laboratory and their candidacidal activities have been found to be inferior to that of Hsn-5 . We were therefore intrigued to explore the candidacidal potency of the full-length MUC7 D1 (51-mer) . Linear solid-phase synthesis of this domain has been accomplished following standard Fmoc chemistry . The problems of partial coupling, owing to the peptide chain length, at several stages of the solid-phase step-by-step synthesis were circumvented either by double-coupling techniques or efficient coupling procedures . The MUC7 D1 peptide was purified to homogeneity by conventional reverse-phase HPLC using two columns connected in series . Secondary structure of the purified peptide was assessed by circular dichroism (CD) spectroscopy in phosphate buffer and trifluoroethanol and compared to that of MUC7 D1-15mer and Hsn-5 . The MUC7 D1 candidacidal activity was assessed against azole-sensitive and azole-resistant C . albicans strains and was found, unlike that of the MUC7 D1-15mer, to be comparable with that of Hsn-5, indicating that in addition to Hsn-5, MUC7 D1 could provide an attractive alternative to the classical antifungal agents . The candidacidal potency of MUC7 D1, like that of MUC7 D1-15mer, and of Hsn-5, appears to be largely dependent on peptide charge, irrespective of alpha-helical structure. Med Mycol, 2000 Oct, 38(5), 363 - 9 Effect of pH, carbon source and K+ on the Na+-inhibited germ tube formation of Candida albicans; Biswas SK et al.; The effect of pH, carbon source and K+ on the Na+ -inhibited germ tube formation of the pathogenic fungus Candida albicans was examined in the arginine-phosphate modified (APM) medium . All C . albicans cells formed germ tubes in APM medium at pH 5.0-9.0 . Na+ inhibited germ tube formation in a concentration dependent manner ranging from 0.2 to 1.0 M, and was further influenced by the pH of the medium . The inhibitory effect of Na+ was lowest at pH 8.0, and germ tube formation ceased at 1.0 M Na+ for any pH (4.0-9.0) . At pH > or = 6.0, non-germ tube-forming cells did not show yeast growth; whereas at pH < or = 5.0, Na+ inhibited only germ tube formation but did not inhibit yeast growth . The inhibitory effect of Na+ was stronger in glucose medium than in galactose medium as carbon source . K+, at 0-0.8 M, had almost no effect on germ tube formation . However, in the presence of Na+, a very low concentration of K+ (0.5 mM) was able to release the cells from Na+ arrest and produced an increase in the rate as well as the percentage of germ tube formation . Intracellular Na+/K+ ratios increased with the increase in extracellular Na+ concentration, whereas the ratios decreased and remained within nontoxic levels when the extracellular K+ concentration was increased. Med Mycol, 2000 Oct, 38(5), 355 - 62 Melaleuca alternifolia (tea tree) oil inhibits germ tube formation by Candida albicans; Hammer KA et al.; The effect of tea tree oil (TTO) on the formation of germ tubes by Candida albicans was examined . Two isolates were tested for germ tube formation (GTF) in the presence of TTO concentrations (% v/v) ranging from 0.25% (1/2 minimum inhibitory concentration {MIC}) to 0.004% (1/128 MIC) . GTF at 4 h in the presence of 0.004 and 0.008% (both isolates) and 0.016% (one isolate) TTO did not differ significantly (P > 0.05) from controls . At all other concentrations at 4 h, GTF differed significantly from controls (P < 0.01) . A further eight isolates were tested for GTF in the presence of 0.031% TTO, and at 4h the mean GTF for all 10 isolates ranged 10.0-68.5% . Two isolates were examined for their ability to form germ tubes after 1 h of pre-exposure to several concentrations of TTO, prior to induction of germ tubes in horse serum . Cells pre-exposed to 0.125 and 0.25% TTO formed significantly fewer germ tubes than control cells at 1 h (P < 0.05), but only those cells pre-exposed to 0.25% differed significantly from control cells at later time points (P < 0.01) . GTF by C . albicans is affected by the presence of, or pre-exposure to, sub-inhibitory concentrations of TTO . This may have therapeutic implications. Int J Antimicrob Agents, 2000 Nov, 16(3), 205 - 9 Activity of voriconazole against Candida albicans and Candida krusei isolated since 1984; Lee JK et al.; With the recent dramatic rise in fluconazole use, there has been an increase in Candida species resistant to that agent . This has led to the clinical development of newer triazoles such as voriconazole that have greater potency and a broader spectrum of activity . We therefore hypothesized that fluconazole-resistant Candida albicans and Candida krusei would be susceptible to voriconazole . Susceptibility testing was performed on 205 isolates of C . albicans collected from 1984 to 1995, and on C . albicans and C . krusei that were identified as fluconazole resistant since 1995 . The anti fungal agents used were amphotericin B, 5-flucytosine, itraconazole, ketoconazole, fluconazole and voriconazole . Three C . albicans and 26 C . krusei isolates had a minimum inhibitory concentration (MIC) >/=20 mg/l and were defined as fluconazole resistant . Of these, 28 isolates were susceptible to </=2.5 mg/l voriconazole, with a mean MIC of 0.78 mg/l . The mean amphotericin B MIC for these same strains was 0.98 mg/l . Only one isolate of C . krusei was relatively resistant to voriconazole with a MIC of 5 mg/l . Of these 29 isolates, there were ten amphotericin-resistant strains of C . krusei (MIC>2 mg/l) but all were susceptible to </=2.5 mg/l voriconazole . The higher potency of voriconazole may be useful in the treatment of fluconazole- and amphotericin-resistant C . albicans and C . krusei. J Adolesc Health, 2000 Dec, 27(6), 384 - 90 The prevalence of anergy in human immunodeficiency virus-infected adolescents and the association of delayed-type hypersensitivity with subject characteristics; Smith Rogers A et al.; PURPOSE: To examine the prevalence of anergy in HIV-infected adolescents and factors associated with its occurrence . METHODS: Anergy was defined as less than 2mm induration to each of three intradermally applied antigens (Candida albicans, tetanus toxoid, and mumps) between 24 and 96 hours in a population of HIV-infected adolescents aged 12-18 at entry in a national multicenter study of HIV disease progression . CD4(+) T-cell counts and plasma HIV-1 RNA were measured in quality controlled laboratories . Factors associated with the probability of anergy were examined with contingency table comparisons, tree-structured classification, and logistic regression analyses . RESULTS: Overall prevalence of anergy in this clinic-based population of 167 was 11% {7% in males and 12% in females (p = 0.57)} . The sole significant predictor of anergy was decreased CD4(+) T-cell count (p = 0.005) . CONCLUSION: The prevalence of anergy is low in this HIV-infected population compared to older infected cohorts . The occurrence of differential rates of anergy in particular age and sex groupings that may be related to intrinsic immunologic differences requires further study. Proc Natl Acad Sci U S A, 2000 Dec 19, 97(26), 14433 - 7 Prevalence of small inversions in yeast gene order evolution; Seoighe C et al.; Gene order evolution in two eukaryotes was studied by comparing the Saccharomyces cerevisiae genome sequence to extensive new data from whole-genome shotgun and cosmid sequencing of Candida albicans . Gene order is substantially different between these two yeasts, with only 9% of gene pairs that are adjacent in one species being conserved as adjacent in the other . Inversion of small segments of DNA, less than 10 genes long, has been a major cause of rearrangement, which means that even where a pair of genes has been conserved as adjacent, the transcriptional orientations of the two genes relative to one another are often different . We estimate that about 1,100 single-gene inversions have occurred since the divergence between these species . Other genes that are adjacent in one species are in the same neighborhood in the other, but their precise arrangement has been disrupted, probably by multiple successive multigene inversions . We estimate that gene adjacencies have been broken as frequently by local rearrangements as by chromosomal translocations or long-distance transpositions . A bias toward small inversions has been suggested by other studies on animals and plants and may be general among eukaryotes. Rev Esp Quimioter, 2000 Sep, 13(3), 314 - 7 {Azole resistance in Candida albicans}; Perea S; The molecular mechanisms of azole resistance in Candida albicans include alterations in the target enzyme (lanosterol 14-demethylase) and overexpression of efflux transporters that decrease the intracellular concentration of the drug . Although the rate of azole resistance in systemic isolates of C . albicans remains very low, resistance to fluconazole appears as an important issue in the management of oropharyngeal candidiasis (OPC) in patients with AIDS . In order to establish the prevalence of resistance to azole antifungal agents in this setting, we investigated the molecular mechanisms of resistance to azoles in highly resistant C . albicans isolates (fluconazole MIC 64 mg/l) from HIV-infected patients with OPC . Antifungal susceptibility testing of serial C . albicans isolates was performed by NCCLS methodology . Strain identity was investigated by DNA-typing techniques . Overexpression of genes encoding lanosterol 14-demethylase (erg11) and efflux transporters (mdr1 and cdr) implicated in the development of resistance was monitored in matched sets of susceptible and resistant isolates . In addition, erg11 genes were PCR-amplified and their nucleotide sequences determined in order to detect point mutations . A combination of different mechanisms of resistance contributed to the development of resistance to fluconazole . The multifactorial character of the azole resistance in C . albicans makes necessary the development of approaches to overcome the problem . Accordingly, new triazoles have been developed; new classes of antifungals are being investigated; and combinations with inhibitors of efflux transporters are being studied. Infect Immun, 2000 Dec, 68(12), 7159 - 61 Repression of hyphal proteinase expression by the mitogen-activated protein (MAP) kinase phosphatase Cpp1p of Candida albicans is independent of the MAP kinase Cek1p; Schroppel K et al.; Cpp1p is a putative mitogen-activated protein (MAP) kinase phosphatase that suppresses Candida albicans hyphal formation at 25 degrees C through its probable substrate, the Cek1p filamentation MAP kinase . Here we report that expression of the serum-induced genes SAP4-6 and HYR1 increased several fold in hyphal forms of a cpp1/cpp1 null mutant, while the rate and extent of hyphal development up to 5 h were normal . Therefore, we provide evidence that Cpp1p represses hyphal gene expression by acting through a Cek1p-independent mechanism . SAP4-6 and HYR1 transcripts were undetectable in a null mutant of another key regulator of filamentation, Efg1p; thus, Efg1p and Cpp1p oppose each other during the expression of these genes in hyphal forms. Infect Immun, 2000 Dec, 68(12), 6848 - 56 Released ATP is an extracellular cytotoxic mediator in salivary histatin 5-induced killing of Candida albicans; Koshlukova SE et al.; Salivary histatins (Hsts) are antifungal peptides with promise as therapeutic agents against candidiasis . Hst 5 kills the fungal pathogen Candida albicans via a mechanism that involves release of cellular ATP in the absence of cytolysis . Here we demonstrate that released ATP has a further role in Hst 5 killing . Incubation of the cells with ATP analogues induced cell death, and addition of the ATP scavenger apyrase to remove extracellular ATP released during Hst 5 treatment resulted in a reduction in cell killing . Experiments using anaerobically grown C . albicans with decreased susceptibility to Hst 5 confirmed that depletion of cellular ATP as a result of ATP efflux was not sufficient to cause cell death . In contrast to Hst-susceptible aerobic cultures, anaerobically grown cells were not killed by exogenously applied ATP . These findings established that Hst binding, subsequent entry into the cells, and ATP release precede the signal for cytotoxicity, which is mediated by extracellular ATP . In a higher-eukaryote paradigm, released ATP acts as a cytotoxic mediator by binding to membrane nucleotide P2X receptors . Based on a pharmacological profile and detection of a C . albicans 60-kDa membrane protein immunoreactive with antibody to P2X(7) receptor, we propose that released ATP in response to Hst 5 activates candidal P2X(7)-like receptors to cause cell death. Infect Immun, 2000 Dec, 68(12), 6833 - 9 Correlation of susceptibility of immature mice to fungal infection (blastomycosis) and effector cell function; Ganer A et al.; Immature mice are highly susceptible to blastomycosis, which is similar to other mycoses and has parallels in humans . The murine susceptibility is noteworthy in that it persists beyond the development of resistance to other, nonfungal pathogens and the maturation of most immune functions . As the susceptibility to blastomycosis appeared to be related to an early event after infection, primary effector cell function was studied . We found that peritoneal inflammatory cells, enriched for neutrophils, from immature (3-week-old) mice killed nonphagocytizable Blastomyces dermatitidis cells less (25%) than did cells from mature (8-week) mice (70%) (P<0.01), a defect intrinsic to the neutrophils . This correlated with an impaired immature cell oxidative burst . Killing of phagocytizable Candida albicans was not significantly different, 73 versus 87% . Thioglycolate-elicited cells were more impaired; killing of B . dermatitidis was insignificant, and killing of C . albicans was more impaired in immature (16% killing) than in mature (45%) cells (P<0.02) . Peripheral blood neutrophils from mature animals killed B . dermatitidis (41%) more than did those from immature animals (10%) (P<0.02); C . albicans was killed efficiently by both . Resting or activated peritoneal macrophages from both types of animals showed no differences in B . dermatitidis killing . These results suggest that the susceptibility of immature mice is related at least in part to the depressed capacity of their neutrophils to kill B . dermatitidis. Infect Immun, 2000 Dec, 68(12), 6777 - 84 Generation of a recombinant 65-kilodalton mannoprotein, a major antigen target of cell-mediated immune response to Candida albicans; La Valle R et al.; A 65-kDa mannoprotein (CaMp65) has long been studied as a major, immunodominant antigen of the human opportunistic pathogen Candida albicans . An expression library of C . albicans was screened with serum from mice immunized with ScMp65 (ScW10), a Saccharomyces cerevisiae recombinant protein of about 48 kDa . This serum recognized the CaMp65 from a cell wall extract of C . albicans . After cloning and sequencing of the relevant C . albicans cDNA, an open reading frame encoding a protein of 379 amino acids was identified . Its deduced amino acid sequence showed regions of identity with all previously characterized tryptic fragments of CaMp65, as well as with the corresponding regions of ScMp65 . A prepeptide of 32 amino acids with signal peptidase and Kex2 cleavage sites as well as a high number of potential O-glycosylation sites but no N-glycosylation sites or GPI anchor were observed in sequence studies of CaMp65 . A putative adhesin RGD sequence was also present in the C-terminal region of the molecule . This triplet was absent in the ScMp65 . The relevant gene (designated CaMP65) was localized to chromosome R of C . albicans as determined by pulse-field gel electrophoresis . Northern blot analysis demonstrated that gene transcription was heat inducible and associated with germ-tube formation by the fungus . A recombinant, His(6)-tagged protein (rCaMp65) was expressed in Escherichia coli under an inducible promoter . After purification by nickel-chelate affinity chromatography, the recombinant product was detected as a 47-kDa protein band in immunoblots with the anti-ScMp65 serum, as well as with CaMp65-specific monoclonal antibodies . Both ScMp65 and CaMp65 were assayed for antigenic stimulation in cultures of peripheral blood mononuclear cells (PBMC) from 10 unselected human donors . While ScMp65 was substantially nonstimulatory, both rCaMp65 and the native CaMp65 were equally able to induce lymphoproliferation of the PBMC from all the donors . In addition, a number of CD4(+) T-cell clones were generated using a C . albicans mannoprotein fraction as an antigenic stimulant . Several of these clones specifically responded to both the native and the recombinant C . albicans Mp65 but not to ScMp65 . Thus, the recombinant Mp65 of C . albicans retains antigenicity and, as such, could be a valid, standardized reagent for serodiagnostic and immunological studies. Infect Immun, 2000 Dec, 68(12), 6712 - 9 Tetracycline-regulatable system to tightly control gene expression in the pathogenic fungus Candida albicans; Nakayama H et al.; Conventional tools for elucidating gene function are relatively scarce in Candida albicans, the most prevalent human fungal pathogen . To this end, we developed a convenient system to control gene expression in C . albicans by the tetracycline-regulatable (TR) promoters . When the sea pansy Renilla reniformis luciferase gene (RLUC1) was placed under the control of this system, doxycycline (DOX) inhibited the luciferase activity almost completely . In the absence of DOX, the RLUC1 gene was induced to express luciferase at a level 400- to 1,000-fold higher than that in the presence of DOX . The same results were obtained in hypha-forming cells . The replacement of N-myristoyltransferase or translation elongation factor 3 promoters with TR promoters conferred a DOX-dependent growth defect in culture media . Furthermore, all the mice infected with these mutants, which are still virulent, survived following DOX administration . Consistently, we observed that the number of these mutant cells recovered from the mouse kidneys was significantly reduced following DOX administration . Thus, this system is useful for investigating gene functions, since this system is able to function in both in vitro and in vivo settings. Antimicrob Agents Chemother, 2000 Dec, 44(12), 3389 - 94 Activities of sordarins in experimental models of candidiasis, aspergillosis, and pneumocystosis; Martinez A et al.; Sordarin derivatives represent a new class of antifungal agents that act as potent inhibitors of fungal protein synthesis and possess a broad spectrum of activity . The in vivo activity of GM193663 and GM237354 was studied in mouse models of disseminated candidiasis and aspergillosis and in a rat model of pneumocystosis . The pharmacokinetic behavior of both sordarin derivatives was studied in mice and rats . In all studies, compounds were administered by the subcutaneous route . After a subcutaneous dose of 50 mg/kg of body weight to mice, the maximum level in serum, area under the concentration-time curve, half-life, and clearance for GM193663 and GM237354 were 51.8 and 23 microg/ml, 79.5 and 46 microg . h/ml, 0.8 and 0.85 h, and 21 and 25 ml/h, respectively . Systemic candidiasis and aspergillosis were established in CD-1 male mice infected with Candida albicans or Aspergillus fumigatus . For systemic candidiasis, compounds were given three times per day for seven consecutive days at 15, 30, 60, or 120 mg/kg/day . GM193663 and GM237354 showed dose-related efficacy against C . albicans, with 50% effective doses, 1 month after infection, of 25.2 and 10.7 mg/kg/dose, respectively . In experimental infections with A . fumigatus, GM237354 was given three times per day at 30, 60, or 120 mg/kg/day for five consecutive days . Animals treated with GM237354 demonstrated irregular responses . The survival of animals treated with GM237354 was 0, 30, and 0% at 30, 60, and 120 mg/kg/day, respectively . The therapeutic efficacy of GM193663 and GM237354 against Pneumocystis carinii was studied in an experimental P . carinii pneumonia (PCP) rat model . After a subcutaneous dose of 10 mg/kg given to rats, the maximum level in serum, area under the concentration-time curve, half-life, and clearance for GM193663 and GM237354 were 6.6 and 7.2 microg/ml, 8.5 and 11.8 microg . h/ml, 0.7 and 0.8 h, and 230 and 133 ml/h, respectively . To induce spontaneous PCP, rats were chronically immunosuppressed with dexamethasone . Infected animals were treated twice daily for 10 days at 0.2, 2, or 10 mg/kg/day . The therapeutic effect was estimated by the reduction in the number of cysts in the lungs of treated versus untreated animals . GM193663 and GM237354 significantly reduced the mean (+/- standard deviation) log number of cysts from 7.6 +/- 0.2 in the untreated group to 4.7 +/- 0.2 and 4.6 +/- 0.1, respectively, when the drugs were administered at a dose of 2 mg/kg/day . The log number of cysts was also reduced in infected animals given lower doses of the compounds (0.2 mg/kg/day) . In summary, GM193663 and GM237354 are new sordarin derivatives that may potentially play a major role in the treatment of candidiasis and PCP . Further testing with Aspergillus in other animal models is warranted. Antimicrob Agents Chemother, 2000 Dec, 44(12), 3310 - 6 Salivary histatin 5 and human neutrophil defensin 1 kill Candida albicans via shared pathways; Edgerton M et al.; Salivary histatins are a family of basic histidine-rich proteins in which therapeutic potential as drugs against oral candidiasis is apparent, considering their potent in vitro antifungal activity and lack of toxicity to humans . Histatin 5 (Hst 5) kills the fungal pathogen Candida albicans via a mechanism that involves binding to specific sites on the yeast cell membrane and subsequent release of cellular ATP in the absence of cytolysis . We explored the killing pathway activated by Hst 5 and compared it to those activated by other antifungal agents . The candidacidal activity of human neutrophil defensin 1 (HNP-1) shared very similar features to Hst 5 cytotoxic action with respect to active concentrations and magnitude of induction of nonlytic ATP efflux, depletion of intracellular ATP pools, and inhibitor profile . Hst 5 and HNP-1 are basic proteins of about 3 kDa; however, they have unique primary sequences and solution structures that cannot explain how these two molecules act so similarly on C . albicans to induce cell death . Our finding that HNP-1 prevented Hst 5 binding to the candidal Hst 5 binding protein suggests that the basis for the overlapping actions of these two naturally occurring antimicrobial proteins may involve interactions with shared yeast components. Antimicrob Agents Chemother, 2000 Dec, 44(12), 3257 - 63 Candidacidal activities of human lactoferrin peptides derived from the N terminus; Lupetti A et al.; In light of the need for new antifungal agents, the candidacidal activities of human lactoferrin (hLF) and synthetic peptides representing the first, hLF(1-11), and second, hLF(21-31), cationic domains of its N terminus were compared . The results revealed that hLF(1-11) was more effective in killing fluconazole-resistant Candida albicans than hLF(21-31) and much more effective than lactoferrin, as determined microbiologically and by propidium iodide (PI) staining . By using hLF(1-11) and various derivatives, it was found that the second and third residues of the N terminus of hLF(1-11) were critical for its candidacidal activity . Detailed investigation to elucidate the mechanism of action of hLF(1-11) revealed a dose-dependent release of ATP by Candida upon exposure to hLF(1-11) . Our observations that sodium azide reduced the PI uptake and candidacidal activity of hLF(1-11) and that, upon exposure to hLF(1-11), the fluorescent dye rhodamine 123 first accumulated inside the mitochondria and later was released into the cytoplasm indicate that the peptide triggers the energized mitochondrion . Furthermore, oxidized ATP, which interferes with the interaction of ATP with its extracellular receptors, blocked the candidacidal action of hLF(1-11), as measured microbiologically and by PI staining . Addition of ATP (or analogues) was not a sufficient stimulus to kill C . albicans or to act synergistically with suboptimal concentrations of the peptide . The main conclusions are that the first two arginines at the N terminus of hLF are critical in the candidacidal activity of hLF(1-11) and that extracellular ATP is essential but not sufficient for the peptide to exert its candidacidal activity. Ginekol Pol, 2000 Sep, 71(9), 971 - 4 {In vitro assessment of the sensitivity of Candida albicans strains isolated from the vagina on basis antimycotics}; Lisiak M et al.; OBJECTIVE: The purpose of the study was to analyses the sensitivity of 73 randomly selected Candida albicans strains isolated from the vagina of pregnant and delivering women against seven basic antimycotics . MATERIALS AND METHODS: The microtest FUNGITEST (Sanofi Diagnostics Pasteur) was applied in assessing the sensitivity of 5-fluorocytosin, amphotericin B, ketoconazol, fluconazol, itraconazol and miconazol and the disk-diffusion method with the use of a Casitone base for nystatin . RESULTS AND CONCLUSIONS: Variations in the sensitivity against drugs have been noted between individual strains of Candida albicans species . The largest number of strains was resistant against ketoconazol--56.16%, and only 10.96% was resistant against nystatin. Ginekol Pol, 2000 Sep, 71(9), 959 - 63 {Yeast species identification in vulvovaginal candidiasis: susceptibility to nystatin}; Lisiak M et al.; OBJECTIVE: The rates of Candida species and susceptibility to nystatin were evaluated . MATERIALS AND METHODS: In the period from January 1, 1998 to December 31, 1998 mycological tests have been carried out for identification of yeast species in the group of pregnant and delivering women hospitalized in the Obstetrics-Gynecological Departments of the Municipal Hospital in Bydgoszcz . We used two commercial media: Albicans ID and CHROMagar Candida . Noted have been 389 positive inoculation results for Candida from vaginal secretions . RESULTS AND CONCLUSIONS: From the total number of 416 differentiated fungus strains decidedly dominant was the species Candida albicans--constituting 81.97% of all strains . The second frequently occurring fungus species was Candida glabrata--11.06% . Further species were C . krusei--2.16%, C . tropicalis--1.20% and C . guilliermondii--1.20% . In 10 cases (2.41%)--in spite of carrying out the laboratory activities that are necessary in such a situation--the species type of tested strains could not be determined . The simultaneous occurrence of two Candida species has been noted in material originating from 27 women (6.94% of cases) . Defining the drug-resistance of 93 Candida species strains against nystatin by means of the disk-diffusion method--it has been started that this drug is highly effective--81.72% of sensitive strains. Curr Microbiol, 2000 Dec, 41(6), 384 - 7 Cloning and characterization of a Candida albicans gene homologous to fructose-1,6-bisphosphatase genes; De la Rosa JM et al.; By sequencing of the DNA adjacent to the Candida albicans SEC61 gene, an open reading frame encoding a polypeptide of 331 amino acids was found . The predicted protein showed a strong homology with the fructose-1,6-bisphosphatase {FbPase} from other organisms, and conserved regions included the catalytic motif found in all known FbPases . Although the cloned gene did not complement the growth failure of a Saccharomyces cerevisiae fbp1 mutant in media with gluconeogenic carbon sources, it was transcribed in the transformants in a fashion that indicates a partial repression by glucose . A similar control on the transcription of this gene and on FbPase activity was found in wild-type C . albicans, where the cloned gene (CaFBP1) was shown to be localized in a single chromosomal locus in the genome. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2000 Nov, 90(5), 651 - 5 The effect of ethylenediamine-tetraacetic acid on Candida albicans; Sen BH et al.; OBJECTIVES: The aim of this study was to evaluate the antifungal effect of ethylenediamine-tetraacetic acid (EDTA) on Candida albicans, comparing it with that of various disinfectants and common antifungal agents . STUDY DESIGN: Two clinical oral isolates and 1 standard strain of C albicans were included in this study . Main contents of the test solutions were sodium hypochlorite, EDTA, chlorhexidine, hexetidine, benzalkonium chloride, povidone-iodine, nystatin, and ketoconazole . The agar diffusion method was used to determine the antifungal effects of the solutions . Zones of inhibition were recorded and the results were analyzed statistically by using a 2-way analysis of variance . RESULTS: EDTA demonstrated the highest antifungal activity in comparison with routine antifungal drugs and all other solutions (P <.0001) . Oral cavity isolate was more resistant to the test solutions (P <.0001) . CONCLUSION: The selection of irrigating and disinfecting solution in root canals of patients with a particularly high incidence of oral candidiasis gains extreme importance . EDTA may be strongly recommended during endodontic therapy of these patients. Fitoterapia, 2000 Dec, 71(6), 690 - 2 Cassine, an antimicrobial alkaloid from Senna racemosa; Sansores-Peraza P et al.; The leaves of Senna racemosa yielded the piperidine alkaloid cassine and an inositol methyl ether . Antimicrobial screening of the compounds revealed antibacterial activity of cassine with minimum inhibitory concentrations of 2.5 mg/ml for Staphylococcus aureus and Bacillus subtilis and 5.0 mg/ml for Candida albicans. Eur J Med Res, 2000 Oct 30, 5(10), 455 - 9 Influence of fluconazole on phagocytosis, oxidative burst and killing activity of human phagocytes . Using a flow cytometric method with whole blood; Baldauf C et al.; OBJECTIVES: The aim of this study was to investigate the influence of fluconazole, an antimycotic on phagocytosis, oxidative burst and killing activity of phagocytes in human whole blood with Candida albicans as a test strain using a flow cytometric method . METHODS: Candida albicans was stained with Calcein AM, a greenfluorescent dye from Bioprobes (Molecular Probes, Inc., P.O . Box 22010, Eugene, OR 97402-0469, USA) . To measure phagocytosis and burst activity diluted monoclonal antibody (CD-13-R-PE, Molecular Probes, Inc., P.O . Box 22010, Eugene, OR 97402-0469, USA) attaching at the surface of granulocytes and monocytes was added as well as Dihydroethidium solution (Molecular Probes, Inc., P.O . Box 22010, Eugene, OR 97402-0469, USA) which changes into red fluorescent Ethidium by oxidation when killing activity takes place . With Ethidium-Homodimer-1-solution (Molecular Probes, Inc., P.O . Box 22010, Eugene, OR 97402-0469, USA) killing activity can be observed . Three different tests, one incubating the Candida for 1- 4 hrs in advance, another incubating whole blood for 1 h, and the third incubating neither yeast nor blood, and a combined main test were carried out . Measurement of phagocytosis, burst- and killing activtiy was performed with a flow cytometric method (Coulter Company, type: Epics-Profile II) . RESULTS: Three different concentrations of fluconazole (5, 20 and 100 microg/ml) show neither decreasing nor increasing influence on phagocytosis and burst activity, irrespective of whether yeasts or phagocytes had been incubated with fluconazole in advance or not . Also after incubating the drug with phagocytes for 1 h, neither an increase nor a decrease of killing activity was observed . A significant increase was, however, found with increasing incubation time of yeasts and fluconazole . - The minimum concentration of fluconazole, just enough to show a significant increase of the killing rate was 1 microg/ml after 3hrs of incubation . No further significant increase was detected when the concentration exceeded 5 microg/ml . CONCLUSION: 1 h incubation of human phagocytes with fluconazole does not have any significant influence on cellular activities . After advanced incubation of Candida a corresponding increase of the intracellular killing rate in phagocytes occurs, probably due to changes of the cytomorphology of yeasts. J Nat Prod, 2000 Oct, 63(10), 1447 - 8 CR377, a new pentaketide antifungal agent isolated from an endophytic fungus; Brady SF et al.; Cultures of endophytic fungi collected in the Guanacaste Conservation Area of Costa Rica were screened for antifungal activity . CR377, a new pentaketide antifungal agent, was isolated from the culture broth of a fungus, CR377 (Fusarium sp.), that showed potent activity against Candida albicans in this assay . The structure of CR377 was established using 1- and 2-D NMR and HRFABMS. J Nat Prod, 2000 Oct, 63(10), 1440 - 3 Lyngbyabellin B, a toxic and antifungal secondary metabolite from the marine cyanobacterium Lyngbya majuscula; Milligan KE et al.; Lyngbyabellin B (1) was isolated from a marine cyanobacterium, Lyngbya majuscula, collected near the Dry Tortugas National Park, Florida . This new cyclic depsipeptide displayed potent toxicity toward brine shrimp and the fungus Candida albicans . The planar structure was deduced using 1D and 2D NMR spectroscopic methods, and the stereochemistry is proposed through a combination of NMR and chiral GC/MS analysis. Mol Cell Biol, 2000 Dec, 20(23), 8696 - 708 Crk1, a novel Cdc2-related protein kinase, is required for hyphal development and virulence in Candida albicans; Chen J et al.; Both mitogen-activated protein kinases and cyclin-dependent kinases play a role in hyphal development in Candida albicans . Using an oligonucleotide probe-based screen, we have isolated a new member of the Cdc2 kinase subfamily, designated Crk1 (Cdc2-related kinase) . The protein sequence of Crk1 is most similar to those of Saccharomyces cerevisiae Sgv1 and human Pkl1/Cdk9 . In S . cerevisiae, CRK1 suppresses some, but not all, of the defects associated with an sgv1 mutant . Deleting both copies of CRK1 in C . albicans slows growth slightly but leads to a profound defect in hyphal development under all conditions examined . crk1/crk1 mutants are impaired in the induction of hypha-specific genes and are avirulent in mice . Consistent with this, ectopic expression of the Crk1 kinase domain (CRK1N) promotes filamentous or invasive growth in S . cerevisiae and hyphal development in C . albicans . The activity of Crk1 in S . cerevisiae requires Flo8 but is independent of Ste12 and Phd1 . Similarly, Crk1 promotes filamentation through a route independent of Cph1 and Efg1 in C . albicans . RAS1(V13) can also activate filamentation in a cph1/cph1 efg1/efg1 double mutant . Interestingly, CRK1N produces florid hyphae in ras1/ras1 strains, while RAS1(V13) generates feeble hyphae in crk1/crk1 strains. J Med Microbiol, 2000 Nov, 49(11), 985 - 91 Heterogeneity of oral isolates of Candida albicans in HIV-positive patients: correlation between candidal carriage, karyotype and disease stage; Capoluongo E et al.; Opportunist infections involving Candida albicans often develop in HIV-positive patients and oral lesions tend to become more frequent as the disease progresses . Previous studies have shown contrasting results concerning the variability of the pulsed-field gel electrophoresis (PFGE) subtypes of C . albicans observed in HIV-positive patients . Carriage of C . albicans was determined by an oral rinse technique; 41 strains of C . albicans (78% serotype A and 22% serotype B) were isolated . There was a direct correlation between candidal load (cfu/ml) and the blood HIV load, whereas there was an inverse correlation with the stage of disease and the CD4 cell counts . The PFGE patterns of isolates were variable with regard to the number and positions of bands . The variability of the band sizes in some run positions showed a Gaussian distribution . Generally, the most frequent size variants were associated with the strains with the highest cfu/ml and lowest CD4 counts (< or =200 cells/microl) . These findings suggest a possible strain selection over time during disease progression, especially in HIV-positive subjects with low CD4 counts. Mol Microbiol, 2000 Nov, 38(3), 435 - 45 The TEA/ATTS transcription factor CaTec1p regulates hyphal development and virulence in Candida albicans; Schweizer A et al.; The temporal and spatial expression of stage-specific genes during morphological development of fungi and higher eukaryotes is controlled by transcription factors . In this study, we report the cloning and functional analysis of the Candida albicans TEC1 (CaTEC1) gene, a new member of the TEA/ATTS family of transcription factors that regulates C . albicans virulence . The promoters of the type 4, 5 and 6 proteinase isogenes (SAP4-6) contain repetitive TEA/ATTS consensus sequence motifs . This finding suggests a possible role for a homologue of Saccharomyces cerevisiae TEC1 during the activation of proteinase gene expression in C . albicans . CaTEC1 is predominantly expressed in the hyphal form of C . albicans . In vitro, serum-induced hyphal formation as well as evasion from MPhi after phagocytosis is suppressed in catec1/catec1 mutant cells . Furthermore, expression of the proteinase isogenes SAP4-6 is no longer inducible in these mutant cells . The deletion of the CaTEC1 gene attenuates virulence of C . albicans in a systemic model of murine candidiasis, although both mutant and revertant cells that were prepared from infected tissues or the vaginal mucosa grew in a hyphal morphology in vivo . CaTEC1 complements the pseudohyphal and invasive growth defect of haploid and diploid S . cerevisiae tec1/tec1 mutant cells and strongly activates the promoter of FLO11, a gene required for pseudohyphal growth . This study provides the first evidence pointing to an essential role for a member of the TEA/ATTS transcription factor family that had so far only been ascribed to function during development as a virulence regulator in microbial pathogenesis. Microbiology, 2000 Nov, 146 ( Pt 11), 2755 - 64 A phosphatidylinositol 3-kinase of Candida albicans influences adhesion, filamentous growth and virulence; Bruckmann A et al.; To determine if cellular functions of the phosphatidylinositol 3-kinase CaVps34p are related to processes governing Candida albicans pathogenicity, both copies of the gene were sequentially disrupted . Homozygous deletion of C . albicans VPS34 resulted in a mutant strain which exhibited defects not only in intracellular vesicle transport processes but also in morphogenesis . The CaVPS34 null mutant was unable to form hyphae on different solid media whilst showing a significantly delayed yeast-to-hyphae transition in liquid media . In addition, the mutant was rendered hypersensitive to temperature and osmotic stresses and had a strongly decreased ability to adhere to mouse fibroblast cells compared to the wild-type strain SC5314 . Finally, evidence was obtained that CaVPS34 is essential for pathogenicity of C . albicans as the CaVPS34 null mutant was shown to be avirulent in a mouse model of systemic infection . C . albicans pathogenicity was restored to a near wild-type degree upon reintroduction of CaVPS34 into the chromosome of the null mutant, demonstrating that the observed avirulence corresponded to the loss of CaVPS34 . Thus, the results suggest that CaVPS34 may serve as a potential target for antifungal drugs. Microbiology, 2000 Nov, 146 ( Pt 11), 2743 - 54 A novel multidrug efflux transporter gene of the major facilitator superfamily from Candida albicans (FLU1) conferring resistance to fluconazole; Calabrese D et al.; Azole resistance in Candida albicans can be mediated by several resistance mechanisms . Among these, alterations of the azole target enzyme and the overexpression of multidrug efflux transporter genes are the most frequent . To identify additional putative azole resistance genes in C . albicans, a genomic library from this organism was screened for complementation of fluconazole hypersusceptibility in Saccharomyces cerevisiae YKKB-13 lacking the ABC (ATP-binding cassette) transporter gene PDR5 . Among the C . albicans genes obtained, a new gene was isolated and named FLU1 (fluconazole resistance) . The deduced amino acid sequence of FLU1 showed similarity to CaMDR1 (formerly BEN(r)), a member of the major facilitator superfamily of multidrug efflux transporters . The expression of FLU1 in YKKB-13 mediated not only resistance to fluconazole but also to cycloheximide among the different drugs tested . The disruption of FLU1 in C . albicans had only a slight effect on fluconazole susceptibility; however, it resulted in hypersusceptibility to mycophenolic acid, thus suggesting that this compound could be a substrate for the protein encoded by FLU1 . Disruption of FLU1 in a background of C . albicans mutants with deletions in several multidrug efflux transporter genes, including CDR1, CDR2 and CaMDR1, resulted in enhanced susceptibility to several azole derivatives . FLU1 expression did not vary significantly between several pairs of azole-susceptible and azole-resistant C . albicans clinical isolates . Therefore, FLU1 seems not to be required for the development of azole resistance in clinical isolates. Nippon Ishinkin Gakkai Zasshi . 2000;41(4):219. Protective antigens and mechanisms of anti-Candida immunity; Antonio C; Life threatening fungal diseases are now frequent in a substantial fraction of the immunocompromised host population . The toxicity and the relative scarcity of efficacious antifungal drugs highlight the need for developing alternative or integrative immunoprophylactic and therapeutic tools; among them the need to develop prophylactic or therapeutic vaccines against candidiasis, a widespread mucosal or deep-seated infection caused primarily by the fungus Candida albicans, are of clear priority . Vaccination is a highly beneficial medical practice, and probably the most cost-effective measure against disease onset and progression . It is based on the use of microbial antigens capable of conferring protection in a susceptible target host . To date, only a handful of Candida albicans antigens have been produced and very few of them have been thoroughly investigated for immunogenicity and protection in experimental models of candidiasis . Thus, approaches to the molecular, biochemical and functional characterization of novel C . albicans encoded molecules are most welcome to improve the perspective of developing in the near future an effective vaccine against C . albicans . Identification of anti-Candida vaccine candidates must take into account the diversity of Candida diseases, the various underlying mechanisms of protection as well as the major immune dysfunctions observed as predisposing factors for disease . Antigens to be considered possible vaccine candidates include members of the aspartyl proteinase (Sap2) family and the 65kDa mannoprotein (MP65) antigen . An additional molecule of C . albicans which has not yet been identified but deserves great consideration as a vaccine candidate is the yeast-killer toxin receptor (KTR) . Initial experimental evidence strongly suggests that the above antigens are able to elicit protective immunity against mucosal and/or systemic candidiasis . A series of molecular, biochemical and immunological studies aimed at validating and strengthening this initial evidence are in progress, with the ultimate goal of producing recombinant or natural antigens that can be assessed for their ability to elicit a protective immunity in animal models and the mechanisms whereby protection is achieved, with emphasis on determination of immune correlates of protection. Nippon Ishinkin Gakkai Zasshi, 2000, 41(4), 211 - 7 {Structural and biochemical characteristics of pathogenic fungus: cell walls, lipids and dimorphism, and action modes of antifungal agents}; Kitajma Y; Cell walls (0.1-0.5 microm in thickness) of dermatophytes, at least Trichophyton mentagrophytes and Epidermophyton floccosum, are built of microfibrils (20 nm in diameter) and matrix embedding the fibrils . These fibrils are composed of chitin (70-80%) and a small amount of glucans, and the matrix is composed of beta-1-3, beta1-6 glucan, glucomannan, galactomannan and peptides . Another characteristic structure is the outermost layer (20-50 nm in thickness) of the cell wall, which consists of hydrophobic protein rodlets . Lipids are thought to play important roles in the regulation of dimorphism and virulence in pathogenic fungus . Generally, the ratio of phospholipid/ergosterol is less than 1 in yeast form and 2-20 in mycelial form cells in Candida albicans and Sporothrix schenckii . During the transition from yeast to mycelial forms, phosphatidylinositol and phosphatidylserine are reduced, whereas phosphatidylcholine increases . Phospho-lipase D is activated on this transition . Phospholipase B is now known to be a virulence factor in C . albicans . Polyene antifungal agents bind to ergosterol in membrane to form complexes, which generate pores and destroy the structures and functions of membrane . Azole antifungal agents inhibit the synthesis of ergosterol leading to deficiency in ergosterol content in membrane, and impair the function of membranes in fungal cells . We show the effects of polyenes on the ultrastructure of fungal plasma membrane and impairment of ionomycin-induced calcium influx in T . mentagrophytes, so that we can compare the differences in mode of actions between these two groups of agents. FEMS Microbiol Lett, 2000 Nov 15, 192(2), 159 - 62 Bactericidal and inhibitory effects of azole antifungal compounds on Mycobacterium smegmatis; Jackson CJ et al.; Azole antifungals are central to therapy and act by inhibiting a cytochrome P450, sterol 14-demethylase and blocking normal sterol synthesis . Our recent identification of a mycobacterial sterol biosynthetic pathway led us to probe the efficacy of a range of these compounds against Mycobacterium smegmatis . Several showed equivalent or greater inhibitory effects to those against Candida albicans, and bactericidal activity was demonstrated for four compounds, clotrimazole, econazole, miconazole and tebuconazole . The major drug used clinically, fluconazole, was ineffective . The results are discussed in the light of the world-wide spread of tuberculosis, including drug-resistant forms and the requirement for new drugs. Pediatrics . 2000 Nov;106(5):E63. Should central venous catheters be removed as soon as candidemia is detected in neonates? Karlowicz MG, Hashimoto LN, Kelly RE Jr, Buescher ES. BACKGROUND: Controversy exists regarding the most appropriate acute management of central venous catheters (CVCs) in neonates with candidemia, with up to two thirds of neonatologists preferring to attempt antifungal therapy without removing CVCs . OBJECTIVE: To determine whether CVCs should be removed as soon as candidemia is detected in neonates . Methods . A cohort study of candidemia and CVC was conducted in infants in a neonatal intensive care unit (NICU) over a 5-year period (1994-1998) . RESULTS: Fifty infants had early-removal CVC (ER-CVC) within 3 days and 54 infants had late-removal CVC (LR-CVC) >3 days after the first positive blood culture for Candida species . All infants were treated with amphotericin B . There was no significant difference between infants in the ER-CVC and LR-CVC groups in terms of gender, ethnicity, birth weight, gestational age, age at candidemia, severity-of-illness scores, distribution of types of CVC, or in the distribution of Candida species causing candidemia . The ER-CVC group had significantly shorter duration of candidemia (median: 3 days; range: 1-14 days), compared with the LR-CVC group (median: 6 days; range: 1-24 days) . The case fatality rate of Candida albicans candidemia was significantly affected by the timing of CVC removal: 0 of 21 (95% confidence interval {CI}: 0-14) infants died in the ER-CVC group in contrast to 9 of 23 (39%; 95% CI: 19-59) in the LR-CVC group . CONCLUSION: Failure to remove CVC as soon as candidemia was detected in neonates was associated with significantly increased mortality in C albicans candidemia and prolonged duration of candidemia regardless of Candida species. J Clin Microbiol, 2000 Nov, 38(11), 4272 - 3 Fungal contamination of food in hematology units; Bouakline A et al.; The prevalence of thermotolerant fungi on non-heat-sterilizable food was determined . Aspergillus spp . were noted in 100% of pepper and regular tea samples, 12 to 66% of fruits, 27% of herbal teas, and 20% of freeze-dried soup samples . All soft cheese samples were contaminated by Geotrichum and yeast (Candida norvegensis) but Candida albicans was never identified. Bioorg Med Chem, 2000 Oct, 8(10), 2487 - 99 Understanding the antifungal activity of terbinafine analogues using quantitative structure-activity relationship (QSAR) models; Gokhale VM et al.; Terbinafine and its analogues, which are a major class of non-azole antifungal agents, are known to act by inhibition of squalene epoxidase enzyme in fungal cells . We have performed a quantitative structure-activity relationship (QSAR) study on a series of 92 molecules using different types of physicochemical descriptors . Inhibitors were divided into five classes depending upon chemical structure . QSAR models were generated for correlation between antifungal activity against Candida albicans using genetic function approximation (GFA) technique . Equations were evaluated using internal as well as external test set predictions . Models generated for all these classes show that steric properties and conformational rigidity of side chains play an important role for the activity . The present QSAR analysis agrees with the results of the previously reported CoMFA study. Eur J Clin Microbiol Infect Dis, 2000 Sep, 19(9), 663 - 70 Epidemiology of yeast colonization in the intensive care unit; Hedderwick SA et al.; In order to investigate the epidemiology of colonization and possible transmission of yeasts among patients and healthcare workers in adult intensive care units (ICUs), 194 patients were followed for a mean of 9 +/- 11 days and 63 healthcare workers were followed for a mean of 132 +/- 52 days . Among the patients, 142 (73%) were colonized by yeast, with Candida albicans being the species most commonly recovered . Most patients (65%) were already colonized with yeast upon admission to the intensive care unit; only 17% became colonized after admission . Persistent colonization occurred in 51 (55%) of 92 patients who had more than three cultures performed; in 75% of them, colonization persisted with the same strain of Candida albicans or Candida glabrata . Bacterial infection in the month preceding entry into the ICU was the only risk factor significantly associated with yeast colonization . Among the healthcare workers, yeasts were isolated from 42 (67%) . Candida albicans was most frequently recovered from the oropharynx (19% of occasions), and Candida parapsilosis was most frequently found on hands (8% of occasions) . Persistent colonization of the oropharynx occurred in only six healthcare workers, and none had persistence of yeasts on hands . In this non-outbreak setting, 5 (4%) of 123 patient/healthcare worker interactions that were linked epidemiologically yielded the same strain of Candida albicans, providing evidence for possible cross-transmission . No similar link was found between healthcare worker-patient interactions and colonization with Candida glabrata or Candida parapsilosis. Yeast, 2000 Nov, 16(15), 1413 - 9 Molecular cloning and characterization of the Candida albicans UBI3 gene coding for a ubiquitin-hybrid protein; Roig P et al.; Using a polyubiquitin cDNA as a probe, we have isolated a clone (pPR3, a pEMBLYe23 derivative plasmid) containing the Candida albicans UBI3 gene coding for a fusion protein . This protein is formed by one ubiquitin subunit fused, at its C-terminus, to an unrelated peptide which is similar to the ribosomal protein encoded by the 3' tail of the Saccharomyces cerevisiae UBI3 gene . Southern blot analysis of chromosomal DNA probed with the 3' non-ubiquitin tail of UBI3 indicated that only one homologous gene is present in the C . albicans genome . Heterelogous expression of pPR3 in a S . cerevisiae ubi3 mutant strain complements the mutant phenotype (slow growth) conferred by the ubi3 defect; this provides direct evidence indicating that the clone contains the C . albicans UBI3 gene Northern blot analysis showed that UBI3 gene is expressed in yeast and germ-tube cells of C . albicans, although the UBI3 mRNA levels in starved yeast cells are below the detection limit; UBI3 mRNA drops to undetectable levels on shifting the temperature of growing yeast cells from 28 degrees C to 42 degrees C . When Northern blot analysis was performed using a specific probe for the polyubiquitin (UBI4) gene, no drop in the mRNA levels was detected following thermal upshift or in starved cells . These results indicate that stress conditions (starvation or thermal upshift) negatively regulate UBI3 expression (transcriptional arrest and/or enhanced mRNA decay), and suggest that UBI4 gene provides ubiquitin during the stress response . In addition, we failed to obtain C . albicans UBI3 null mutant cells by sequential disruption of both alleles using the hisG::URA3::hisG ('ura-blaster') cassette, suggesting that null mutants cells may be unable to grow on selective media after transformation. Chemotherapy, 2000 Nov-Dec, 46(6), 395 - 401 In vitro susceptibility of Candida dubliniensis to current and new antifungal agents; Quindos G et al.; BACKGROUND: Candida dubliniensis is a recently described Candida species closely related to Candida albicans, which has been associated with oral candidiasis in HIV-infected patients . Fluconazole-resistant strains of C . dubliniensis are easily obtained in vitro and this fact could be a complication if this resistance develops during treatment with this drug . METHODS: In the present study, the in vitro antifungal susceptibilities of 36 C . dubliniensis clinical isolates and culture strains to current and new antifungal agents, such as amphotericin B (AMB), amphotericin B lipid complex (ABLC), amphotericin B colloidal dispersion (ABCD), 5-fluorocytosine (5FC), fluconazole (FLC), itraconazole (ITC), ketoconazole (KTC), liposomal amphoteri- cin B (LAMB), liposomal nystatin (LNYT), LY303366 (LY), SCH56592 (SCH), and voriconazole (VRC), were determined according to the National Committee for Clinical Laboratory Standards M27-A broth microdilution method for yeasts . RESULTS: Most isolates of C . dubliniensis were susceptible to both new and current antifungal drugs, with 75.9% isolates susceptible to KTC, 86.2% to FLC and to ITC, and approximately 100% to the other antifungal agents tested . The cross-resistance phenotypes are detailed . Four isolates were resistant (MIC > or =64 microg/ml) to FLC . These 4 isolates were also resistant to KTC, and 3 of them were also resistant to ITC (MIC > or =1 microg/ml for both agents) . However, these isolates were highly susceptible to 5FC and all polyene formulations (AMB, ABLC, ABCD, LAMB, and LNYT), triazole (SCH and VRC) and echinocandin (LY) antifungal agents . CONCLUSION: The new liposomal and lipidic formulations of AMB, LNYT, and the new triazoles and echinocandins may provide new alternatives to FLC for the treatment of infections by C . dubliniensis . Acta Biochim Pol, 2000, 47(2), 481 - 6 QSARs of some novel isosteric heterocyclics with antifungal activity; Yalcin I et al.; QSAR analysis of a set of previously synthesized 2,5,6-trisubstituted benzoxazole, benzimidazole and 2-substituted oxazolo(4,5-b)pyridine derivatives tested for growth inhibitory activity against Candida albicans, was performed by using the computer-assisted multiple regression procedure . The activity contributions for either heterocyclic ring systems or substituent effects of these compounds were determined from the correlation equation and the predictions for the lead optimization were described . The resulting QSAR revealed that the oxazolo(4,5-b)pyridine ring system with the substitution of a benzyl moiety at position 2 was the most favourable structure among the heterocyclic nuclei . Moreover, the fifth position in the fused ring system is found more significant than the other positions in improving the activity. Br J Biomed Sci, 2000, 57(3), 241 - 9 Adherence mechanisms of Candida albicans; Cotter G et al.; The yeast Candida albicans is an opportunistic fungal pathogen that is capable of inducing a range of superficial and systemic diseases in the immunocompromised host . Although it displays a variety of virulence factors, one--the ability to adhere to host tissue--is considered essential in the early stages of colonisation and tissue invasion . Adherence is achieved by a combination of specific (ligand-receptor interactions) and non-specific (electrostatic charge, van der Waals forces) mechanisms which allow the yeast to attach to a wide range of tissue types and inanimate surfaces . Conventional methods for treating disease cause by C . albicans rely upon the use of antifungal drugs designed to kill the yeast or arrest its growth . An alternative approach, aimed at disrupting the adherence of the yeast to host tissue in cases of superficial infection, may have potential for controlling disease, particularly in situations where the unattached fungal cell can be removed from the affected site, either by the flushing action of the oropharynx or by the production of mucus in the vagina. Hosp Med, 2000 Sep, 61(9), 605 - 9 Epidemiology; Ellis M et al.; Invasive fungal infections have emerged as important causes of hospital related morbidity and mortality . They are increasingly seen in patients not previously considered at risk, e.g . patients on an intensive care unit . Candida albicans and Aspergillus spp . are the most common pathogens, posing challenges in epidemiology, control and treatment. Ned Tijdschr Geneeskd . 2000 Oct 7;144(41):1953. {Diagnostic image (6) (Pustulous skin lesions)}; van der Meer JW et al.; During chemotherapy a 35-year-old woman developed pustulous skin lesions . Biopsies of these lesions showed Candida albicans . Blood cultures were positive for C . albicans . The patient died. Life Sci, 2000 Sep 22, 67(18), 2247 - 56 Candida albicans and Saccharomyces cerevisiae cell wall mannans produce fever in rats: role of nitric oxide and cytokines; Ataoglu H et al.; Mannan components of C . albicans (5 mg/kg, i.p.) and S . cerevisiae (2.5 mg/kg, i.p.) cell walls produced pyrogenic responses which were completely inhibited by indomethacin (5 mg/kg, s.c.) pretreatment in rats . A non-selective NOS inhibitor, L-NAME (10 mg/kg, s.c.), also inhibited the pyrogenic effectiveness of C . albicans mannan, whereas it was ineffective on the fever induced by S . cerevisiae mannan . A selective elevation in the serum TNF-alpha levels was observed at the initial phase of the fever due to S . cerevisiae mannan, whereas there was no significant change on the serum levels of TNF-alpha, IL-1beta and IFN-gamma during the latent period or at the initial phase of the fever induced by C . albicans mannan . Injections of N-linked and/or O-linked oligomannosides of the either mannan did not cause any significant change in the body temperature and serum cytokine levels . These data suggest that the mannan components of C . albicans and S . cerevisiae cell walls produce a prostaglandin-dependent fever in rats . The initial signal for fever seems to be different for each mannan . Data also indicate that integrity of the mannans is necessary for the pyrogenic response. Int J Med Microbiol, 2000 Mar, 290(1), 97 - 104 Assessment of genetic relatedness of vaginal isolates of Candida albicans from different geographical origins; Pinto de Andrade M et al.; PCR fingerprinting with single non-specific primers was used to type vaginal isolates of C . albicans from Portugal, Angola, Madagascar, and two regions of Germany (Berlin and Munich) . In addition to analysing isolates that exhibited the normal biotype of C . albicans, the study included atypical strains that failed to assimilate glucosamine and N-acetylglucosamine, which were isolated from women in Angola and Madagascar . A total of 212 strains of C . albicans were studied, representing 87 different multi-locus genotypes . The genotypes of strains from each geographical population were highly similar but not identical . There was one exception: a strain from Portugal grouped with the typical strains from Angola . The typical and especially the atypical populations from Africa displayed less genotype variation than the populations from Europe . The Portuguese samples exhibited the greatest genotypic heterogeneity . Distance analysis (UPGMA) revealed a statistically weak correlation between genotype and geographical origin of the C . albicans isolates. Mycopathologia, 1999, 147(3), 117 - 20 New antifungal agents that inhibit the growth of Candida species: dichlorinated 8-quinolinols; Lentz DL et al.; Five dichlorinated 8-quinolinols (2,5- 5,6-, 3,5-, 3,7-, and 4,5-dichloro-8-quinolinol) were tested against Candida albicans and C . tropicalis in Sabouraud dextrose broth with and without bovine serum . The 5,6-, 3,5-, and 3,7-dichloro-8-quinolinols proved to be more effective than the control, 5-fluorocytosine . In cytotoxicity tests employing baby hamster kidney (BHK) cells, all test agents proved to be more cytotoxic than the control . However, the minimum inhibitory concentration (MIC) of 3,5-dichloro-8-quinolinol to both fungi was only one tenth the cytotoxic dose, suggesting that the compound may be useful as a topical or systemic antifungal agent. FEMS Microbiol Lett, 2000 Nov 1, 192(1), 27 - 31 Complement activation in SCID and nude mice is related to severity of tissue inflammation in the Candida mastitis model; Guhad FA et al.; A small animal model of localised candidiasis is needed for the evaluation of new antifungal compounds . Mammary glands of immunocompetent (BALB/cJ) and immunodeficient (SCID and athymic nude) mice were infected with a wild-type of Candida albicans . Some of the animals were treated with amphotericin B (AmB) while others were treated with saline and acted as controls . The histologic changes of infected mammary gland tissues and a number of other organs were evaluated . Complement (C) activation was analysed by immunoelectrophoretic quantification of molecules with C3c epitopes (C3, C3b, iC3b, and C3c) in serum . In all animals the organisms were confined to the mammary glands . Serum C3c levels were significantly higher (P<0.05) in infected untreated BALB/cJ and SCID mice, which also had severe mammary gland tissue inflammation, compared with control mice treated with AmB (4 mg kg(-1) i.p . once daily for 4 days) . Both treated and control nude mice showed less tissue inflammation compared to BALB/cJ and SCID mice, and revealed insignificant activation of the complement system . It is concluded that innate immune response is important in the control of candidiasis and that the murine mastitis model is useful for immunopathological studies as well as evaluation of potential antifungal compounds. Mycoses, 2000 Sep, 43(7-8), 273 - 9 Changes of virulence factors accompanying the phenomenon of induced fluconazole resistance in Candida albicans; Fekete-Forgacs K et al.; We investigated a fluconazole-sensitive (MICflu = 5 micrograms ml-1) clinical isolate and a fluconazole-resistant (MICflu > 80 micrograms ml-1) laboratory mutant Candida albicans strain developed from the sensitive one . We studied putative virulence factors including germination, adherence ability to either buccal epithelial cells or acrylate surface, the secreted aspartic proteinase, and the extracellular phospholipase activity of the two strains as well as their growth . The fluconazole-resistant strain proved to be superior to the original strain in all the virulence traits tested . The higher virulence of the fluconazole-resistant strain was also supported by a mouse model . These results suggest that the development of fluconazole resistance can be accompanied by serious morphological and physiological changes: several putative virulence traits, moreover the in vivo virulence can increase simultaneously. Antimicrob Agents Chemother, 2000 Nov, 44(11), 2985 - 90 Genetic analysis of azole resistance in the Darlington strain of Candida albicans; Kakeya H et al.; High-level azole resistance in the Darlington strain of Candida albicans was investigated by gene replacement in C . albicans and expression in Saccharomyces cerevisiae . We sequenced the ERG11 gene, which encodes the sterol C(14)alpha-demethylase, from our copy of the Darlington strain . Both alleles contained the histidine for tyrosine substitution at position 132 (Y132H) reported in Darlington by others, but we also found a threonine-for-isoleucine substitution (I471T) not previously reported in the C . albicans ERG11 . The encoded I471T change in amino acids conferred azole resistance when overexpressed alone and increased azole resistance when added to the Y132H amino acid sequence in an S . cerevisiae expression system . Replacement of one copy of ERG11 in an azole-susceptible strain of C . albicans with a single copy of the Darlington ERG11 resulted in expression of the integrated copy and a modest increase in azole resistance . The profound azole resistance of the Darlington strain is the result of multiple mutations. Antimicrob Agents Chemother, 2000 Nov, 44(11), 2932 - 8 Fluconazole plus cyclosporine: a fungicidal combination effective against experimental endocarditis due to Candida albicans; Marchetti O et al.; Recent observations demonstrated that fluconazole plus cyclosporine (Cy) synergistically killed Candida albicans in vitro . This combination was tested in rats with C . albicans experimental endocarditis . The MICs of fluconazole and Cy for the test organism were 0.25 and >10 mg/liter, respectively . Rats were treated for 5 days with either Cy, amphotericin B, fluconazole, or fluconazole-Cy . Although used at high doses, the peak concentrations of fluconazole in the serum of rats (up to 4.5 mg/liter) were compatible with high-dose fluconazole therapy in humans . On the other hand, Cy concentrations in serum (up to 4.5 mg/liter) were greater than recommended therapeutic levels . Untreated rats demonstrated massive pseudohyphal growth in both the vegetations and the kidneys . However, only the kidneys displayed concomitant polymorphonuclear infiltration . The therapeutic results reflected this dissociation . In the vegetations, only the fungicidal fluconazole-Cy combination significantly decreased fungal densities compared to all groups, including amphotericin B (P < 0.0001) . In the kidneys, all regimens except the Cy regimen were effective, but fluconazole-Cy remained superior to amphotericin B and fluconazole alone in sterilizing the organs (P < 0.0001) . While the mechanism responsible for the fluconazole-Cy interaction is hypothetical, this observation opens new perspectives for fungicidal combinations between azoles and other drugs. Curr Microbiol, 2000 Mar, 40(3), 145 - 8 Inhibition of germ tube formation by Candida albicans by local anesthetics: an effect related to ionic channel blockade; Rodrigues AA et al.; Formation of germ tubes by Candida albicans has been assumed as a putative virulence factor . Local anesthetics (LAs), e.g., lidocaine and bupivacaine, are known to inhibit germ tube formation . The study confirmed this observation for the novel drug ropivacaine, although it was less potent than the former two drugs . Hypothesizing that the effect is due to blockading ionic channels, we exposed Candida albicans to selective calcium blockers, i.e., nifedipine and verapamil, and to a general blocker of ionic channels, i.e., lanthanum . All blockers inhibited germ tube formation . The effect was dose-dependent and pH-independent . Addition of calcium reverted the effect of the blockers as well as the effect of lidocaine and ropivacaine . The study suggests that the inhibitory effect of LAs on germ tube formation by C . albicans is due to blockade of ionic channels, particularly calcium channels . Therefore, LAs can affect morphology and probably also the pathogenesis of C . albicans. Microbios, 2000, 103(404), 53 - 7 The effect of azidothymidine on germ tube formation in Candida albicans; Monno R et al.; Candida albicans have a marked propensity to cause infections in AIDS patients . A virulent trait of C . albicans is the yeast-hypha transition (Y-->H) which is influenced, in vitro and in vivo, by several factors . Since azidothymidine (AZT) is used in HIV-positive patients, the effect, in vitro, of different concentrations of AZT on C . albicans Y-->H transition was evaluated . C . albicans isolated from HIV-negative and HIV-positive patients were used and strains of C . tropicalis isolated from HIV-positive patients were also tested . AZT concentrations from 0.01 microg/ml to 10 microg/ml did not have any influence on the Y-->H transition, whereas 100 microg/ml AZT significantly inhibited the germ tube formation . AZT did not influence the formation of pseudohyphae in C . tropicalis . It is suggested that C . albicans infection observed in HIV-positive patients was not influenced by AZT therapy, because at currently used dosages, the Y-->H transition was not expected to increase. J Biol Chem, 2001 Jan 19, 276(3), 1857 - 64 Epub 2000 Oct 16. Characterization of the mRNA capping apparatus of Candida albicans; Schwer B et al.; The mRNA capping apparatus of the pathogenic fungus Candida albicans consists of three components: a 520- amino acid RNA triphosphatase (CaCet1p), a 449-amino acid RNA guanylyltransferase (Cgt1p), and a 474-amino acid RNA (guanine-N7-)-methyltransferase (Ccm1p) . The fungal guanylyltransferase and methyltransferase are structurally similar to their mammalian counterparts, whereas the fungal triphosphatase is mechanistically and structurally unrelated to the triphosphatase of mammals . Hence, the triphosphatase is an attractive antifungal target . Here we identify a biologically active C-terminal domain of CaCet1p from residues 202 to 520 . We find that CaCet1p function in vivo requires the segment from residues 202 to 256 immediately flanking the catalytic domain from 257 to 520 . Genetic suppression data implicate the essential flanking segment in the binding of CaCet1p to the fungal guanylyltransferase . Deletion analysis of the Candida guanylyltransferase demarcates an N-terminal domain, Cgt1(1-387)p, that suffices for catalytic activity in vitro and for cell growth . An even smaller domain, Cgt1(1-367)p, suffices for binding to the guanylyltransferase docking site on yeast RNA triphosphatase . Deletion analysis of the cap methyltransferase identifies a C-terminal domain, Ccm1(137-474)p, as being sufficient for cap methyltransferase function in vivo and in vitro . Ccm1(137-474)p binds in vitro to synthetic peptides comprising the phosphorylated C-terminal domain of the largest subunit of RNA polymerase II . Binding is enhanced when the C-terminal domain is phosphorylated on both Ser-2 and Ser-5 of the YSPTSPS heptad repeat . We show that the entire three-component Saccharomyces cerevisiae capping apparatus can be replaced by C . albicans enzymes . Isogenic yeast cells expressing "all-Candida" versus "all-mammalian" capping components can be used to screen for cytotoxic agents that specifically target the fungal capping enzymes. J Enzyme Inhib, 2000, 15(5), 429 - 41 N3-oxoacyl derivatives of L-2,3-diaminopropanoic acid and their peptides; novel inhibitors of glucosamine-6-phosphate synthase; Andruszkiewicz R et al.; Novel inhibitors 1-4 of glucosamine-6-phosphate synthase from Candida albicans have been designed based on acylation of the N3 amino group of L-2,3-diaminopropanoic acid with the corresponding ketoacids . These inhibitors have been shown to alkylate the fungal enzyme in a time-dependent manner . Compound 3 containing trans-beta-benzoyl acrylic acid as an acyl residue was found to be the most potent inhibitor in the series . Dipeptides composed of the active inhibitors and norvaline demonstrated potent antifungal activity against selected strains of Candida spp . and Saccharomyces cerevisiae . Their activity was reversed upon addition of N-acetylglucosamine to the medium. Shock, 2000 Sep, 14(3), 278 - 83 Phagocytosis of Candida albicans induces apoptosis of human neutrophils; Rotstein D et al.; Neutrophil-mediated inflammation is terminated through the programmed cell death or apoptosis of the neutrophil, a process that can be inhibited by soluble mediators released during an inflammatory response . It has been reported, however, that the phagocytosis of intact bacteria can accelerate apoptosis . We evaluated the effects of the phagocytosis of a common nosocomial pathogen, Candida albicans, on the expression of apoptosis . Phagocytosis of killed Candida induced a dose-dependent increase in the apoptosis of normal neutrophils after 18 h of in vitro culture, from 40.7+/-9.1% to 81.7+/-4.5%, while supernatants from neutrophil:Candida co-cultures actually inhibited apoptosis . Induction of apoptosis was not dependent on phagocytosis, since opsonization of yeast with serum failed to increase apoptosis, while inhibition of phagocytosis with latrunculin B resulted in a slightly increased apoptotic rate . Increased apoptosis induced by Candida was associated with increased activity of the membrane-associated apoptotic enzyme, caspase 8, and with increased expression of the active form of the key executioner caspase, caspase 3 . Increased apoptosis was associated with depletion of intracellular glutathione (GSH), and could be inhibited by the addition of exogenous GSH . These data demonstrate an important physiologic role for host-pathogen interactions in the resolution of inflammation and suggest that the response to an invading pathogen is an important stimulus to the restoration of normal immunologic homeostasis. Biochem Biophys Res Commun, 2000 Oct 5, 276(3), 1278 - 85 Solution structure of a designed amphipathic antimicrobial synthetic peptide, PGAa; Hong E et al.; A designed peptide, PGAa showed an excellent antifungal activity as well as an efficient bactericidal activity toward gram-positive, especially in the pathogenic yeast Candida albicans 28838 . The solution structures of PGAa have been determined both in 40% TFE/water solution and DPC micelle by CD and NMR spectroscopy . Based on NOEs, vicinal coupling constants, backbone amide exchange rates, and chemical shift indices, PGAa formed a long amphipathic alpha-helical conformation in both TFE and DPC micelle environments, spanning the residues Ile(2)-Ala(19) in TFE and Lys(5)-Ala(19) in DPC micelle, respectively . Solution structures suggested that the hydrophobic residues would interact with the fatty acyl chains of the lipid bilayer, while the positively charged side-chains exposed to aqueous environments . Therefore, we conclude that the alpha-helical structure as well as the highly amphiphatic nature of PGAa peptide may play a critical role in its antimicrobial activity as well as selectivities in different species . Diagn Microbiol Infect Dis, 2000 Sep, 38(1), 17 - 20 Treatment with oxoglaucine can enhance host resistance to Candida albicans infection of mice with adjuvant arthritis; Ivanovska N et al.; The alkaloid oxoglaucine reduced CD4+ cell clones in adult mice and decreased CD4+, CD8+ and Ig+ levels in newborn mice . It prevented the increase of CD8+ and Ig+ clones induced by Candida albicans (C . albicans) in adult mice . TNF-alpha serum accumulation was inhibited by oxoglaucine in C . albicans infection and adjuvant arthritis . Treatment with oxoglaucine of arthritic mice, followed by inoculation with C . albicans enhanced the host resistance against the pathogen. J Infect, 2000 Sep, 41(2), 176 - 8 Medical treatment of a pacemaker endocarditis due to Candida albicans and to Candida glabrata; Roger PM et al.; We describe a case of pacemaker infection due to two fungal species: Candida albicans and C . glabrata . Transthoracic echocardiography showed a large vegetation on the intraventricular wires . Because of severe underlying diseases, surgery was believed to be contraindicated . The patient was treated using high dose of fluconazole, resulting in clinical improvement and negative blood cultures . However, 2 months later, the patient underwent a fatal stroke . At autopsy, a large vegetation was found only all along the wires . Postmortem culture of the infected material was positive for both C . albicans and C . glabrata . J Infect Dis, 2000 Nov, 182(5), 1479 - 85 Epub 2000 Oct 09. Growth inhibition of Candida by human oral epithelial cells; Steele C et al.; Oropharyngeal candidiasis (OPC) caused by Candida albicans is a significant problem in human immunodeficiency virus (HIV)-infected persons . Recognizing the paucity of information on innate and/or adaptive mucosal host defenses against C . albicans, we recently reported that human and nonhuman primate and mouse vaginal epithelial cells inhibit the growth of C . albicans in vitro . In the present study, oral epithelial cells collected from saliva of healthy volunteers and a purified oral epithelial cell line were found to inhibit blastoconidia and/or hyphal growth of several Candida species . Cell contact was a strict requirement for the epithelial cell anti-Candida activity; neither saliva nor culture supernatants alone inhibited Candida growth, and addition of saliva to the coculture did not modulate the epithelial cell activity . Finally, epithelial cell anti-Candida activity was significantly lower in HIV-infected persons with OPC . Together, these results suggest that oral epithelial cells may play a role in innate resistance against OPC. Klin Med (Mosk), 2000, 78(8), 47 - 51 {Infectious endocarditis in intravenous drug abusers}; Demin AA et al.; Clinical course of infectious endocarditis (IE) was analysed for 43 intravenous drug abusers . 42 of them had primary IE, one patient--secondary . Acute course and high activity of the disease were registered in 86% of the patients . IE was provoked by Staphylococcus aureus (50%), Staphylococcus epidermidis 920%), Staphylococcus haemolyticus (11%), E . coli (8%), Pseudomonas aeruginosa (2%), Candida albicans (2%), mixed microflora (7%) . Vegetations were detected on the tricuspid, mitral and aortic valves (52, 23 and 19%, respectively), on more than one valve (6%) . Pneumonia, pleuricy, hydrothorax, enlargement of the liver, spleen, nephritis and anemia were found in 76, 44, 9, 100, 75, 70 and 88% of the patients, respectively . Cardiac failure aggravated the disease in half of the patients, lethality was 18% . Thus, IE in intravenous drug abusers is characterized by a primary form, acute active course, prevalent damage to the tricuspid valve, polyorganic involvement, high lethality . IE cure in such patients is feasible only in adequate antibacterial therapy, timely surgical correction and giving up drug abuse. Sao Paulo Med J, 2000 Sep 7, 118(5), 131 - 8 Epidemiology of bloodstream infections at a cancer center; Velasco E et al.; CONTEXT: Cancer patients are at unusually high risk for developing bloodstream infections (BSI), which are a major cause of in-hospital morbidity and mortality . OBJECTIVE: To describe the epidemiological characteristics and the etiology of BSI in cancer patients . DESIGN: Descriptive study . SETTING: Terciary Oncology Care Center . PARTICIPANTS: During a 24-month period all hospitalized patients with clinically significant BSI were evaluated in relation to several clinical and demographic factors . RESULTS: The study enrolled 435 episodes of BSI (349 patients) . The majority of the episodes occurred among non-neutropenic patients (58.6%) and in those younger than 40 years (58.2%) . There was a higher occurrence of unimicrobial infections (74.9%), nosocomial episodes (68.3%) and of those of undetermined origin (52.8%) . Central venous catheters (CVC) were present in 63.2% of the episodes . Overall, the commonest isolates from blood in patients with hematology diseases and solid tumors were staphylococci (32% and 34.7%, respectively) . There were 70 episodes of fungemia with a predominance of Candida albicans organisms (50.6%) . Fungi were identified in 52.5% of persistent BSI and in 91.4% of patients with CVC . Gram-negative bacilli prompted the CVC removal in 45.5% of the episodes . Oxacillin resistance was detected in 26.3% of Staphylococcus aureus isolates and in 61.8% of coagulase-negative Staphylococcus . Vancomycin-resistant enterococci were not observed . Initial empirical antimicrobial therapy was considered appropriate in 60.5% of the cases . CONCLUSION: The identification of the microbiology profile of BSI and the recognition of possible risk factors in high-risk cancer patients may help in planning and conducting more effective infection control and preventive measures, and may also allow further analytical studies for reducing severe infectious complications in such groups of patients. Annu Rev Microbiol, 2000, 54, 463 - 98 Recent developments in molecular genetics of Candida albicans; De Backer MD et al.; The frequency of opportunistic infections caused by the fungus Candida albicans is very high and is expected to continue to increase as the number of immunocompromised patients rises . Research initiatives to study the biology of this organism and elucidate its pathogenic determinants have therefore expanded significantly during the last 5-10 years . The past few years have also brought continuous improvement in the techniques to study gene function by gene inactivation and by regulated gene expression and to study gene expression and protein localization by using gene reporter systems . As steadily more genomic sequence information from this human fungal pathogen becomes available, we are entering a new era in antimicrobial research . However, many of the currently available molecular genetics tools are poorly adapted to a genome-wide functional analysis in C . albicans, and further development of these tools is hampered by the asexual and diploid nature of this organism . This review outlines recent advances in the development of molecular tools for functional analysis in C . albicans and summarizes current knowledge about the genomic and genetic variability of this important human fungal pathogen. Fungal Genet Biol, 2000 Jul, 30(2), 155 - 62 Filamentous growth of Saccharomyces cerevisiae is regulated by manganese; Asleson CM et al.; The Candida albicans INT1 gene is a virulence factor that contributes to both adhesion and filamentous growth of the fungus . Expression of INT1 in the budding yeast Saccharomyces cerevisiae directs both adhesion and filamentous growth . Because Int1p contains two predicted divalent cation-binding motifs, we asked whether divalent cations are important for the role of Int1p in filament formation . In this study, we found that INT1-induced filamentous growth (I-IFG) is sensitive to the divalent cation chelator EDTA and that this EDTA sensitivity can be ameliorated by the addition of Mn(2+), but not Mg(2+) or Ca(2+) ions . The addition of MnCl(2) restored both the proportion of cells forming filaments and the length of filaments formed . Expression of INT1 in S . cerevisiae mutants that reduce the intracellular concentration of Mn(2+) did not affect I-IFG . Interestingly, the Mn(2+) dependence of I-IFG is not dependent upon the presence of the putative divalent cation-binding domains found in INT1 . Rather, we found that polarized growth induced by mutations in CDC12 and CLA4 or by expression of excess SWE1 was also sensitive to EDTA treatment and was restored by the addition of MnCl(2) but not by the addition of CaCl(2) . Thus, our results suggest that in S . cerevisiae polarized growth is dependent upon the presence of Mn(2+) ions . Fungal Genet Biol, 2000 Jul, 30(2), 127 - 33 Biosynthesis of glycoproteins in Candida albicans: activity of mannosyl and glucosyl transferases; Arroyo-Flores BL et al.; The enzymes dolichol phosphate glucose synthase and dolichol phosphate mannose synthase (DPMS), which catalyze essential steps in glycoprotein biosynthesis, were solubilized and partially characterized in Candida albicans . Sequential incubation of a mixed membrane fraction with increasing concentrations of Nonidet P-40 released a soluble fraction that transferred glucose from UDP-Glc to dolichol phosphate glucose and minor amounts of glucoproteins in the absence of exogenous dolichol phosphate . Studies with the soluble fraction revealed that some properties were different from those previously determined for the membrane-bound enzyme . Accordingly, the soluble enzyme exhibited a twofold higher affinity for UDP-Glc and a sixfold higher affinity over the competitive inhibitor UMP, and the transfer reaction was fourfold more sensitive to inhibition by amphomycin . On the other hand, a previously described protocol for the solubilization of mannosyl transferases that rendered a fraction exhibiting both DPMS and protein mannosyl transferase (PMT) activities operating in a functionally coupled reaction was modified by increasing the concentration of Nonidet P-40 . This resulted in a solubilized preparation enriched with DPMS and nearly free of PMT activity which remained membrane bound . DPMS solubilized in this manner exhibited an absolute dependence on exogenous Dol-P . Uncoupling of these enzyme activities was a fundamental prerequisite for future individual analysis of these transferases . Pediatrics, 2000 Oct, 106(4), 712 - 8 When to suspect fungal infection in neonates: A clinical comparison of Candida albicans and Candida parapsilosis fungemia with coagulase-negative staphylococcal bacteremia; Benjamin DK Jr et al.; OBJECTIVES: To determine the epidemiology of candidemia in our neonatal intensive care unit; to compare risk factors, clinical presentation, and outcomes for neonates infected with Candida albicans, Candida parapsilosis, and coagulase-negative staphylococcus (CoNS); and to suggest a rational approach to empiric antifungal therapy of neonates at risk for nosocomial infection . DESIGN: Retrospective chart review of all neonatal intensive care unit patients with systemic candidiasis or CoNS infection between January 1, 1995 and July 31, 1998 at Duke University Medical Center . RESULTS: Fifty-one patients were reviewed . Nine of 19 patients infected with C parapsilosis and 5 of 15 patients infected with C albicans died of fungemia . Seventeen neonates had >2 positive cultures for CoNS obtained within 96 hours and 1 died . There was no statistically significant difference in birth weight, gestational age, or age at diagnosis between patient groups; however, candidemic patients had a sevenfold higher mortality rate . Before diagnosis, candidemic patients had greater exposure to systemic steroids, antibiotics, and catecholamine infusions . Of the 51 patients, 32 received third-generation cephalosporins in the 2 weeks before diagnosis and 19 did not . Twenty-nine of the 32 who were treated with third-generation cephalosporins subsequently developed candidemia, while candidemia occurred in only 5 of 19 patients who were not treated with cephalosporins . At the time of diagnosis, candidemic patients were more likely to have required mechanical ventilation and were less likely to be tolerating enteral feeding . Multivariate clustered logistic regression analysis revealed that candidemic patients had more exposure to third-generation cephalosporins . Once the clinician was notified of a positive blood culture for Candida, patients infected with C parapsilosis retained their central catheters longer than patients infected with C albicans . CONCLUSIONS: In this retrospective review, we were able to identify aspects of the clinical presentation and medication history that may be helpful in differentiating between candidemia and CoNS bacteremia . Those key features may be used by clinicians to initiate empiric amphotericin B therapy in premature neonates at risk for nosocomial infections . Prolonged use of third-generation cephalosporins was strongly associated with candidemia . There was no statistically significant difference in the morbidity and mortality between patients infected with C parapsilosis and those infected with C albicans . Observed delays in removal of the central venous catheter may have contributed to finding a mortality rate from C parapsilosis that was higher than was previously reported. J Clin Microbiol, 2000 Oct, 38(10), 3696 - 704 Differentiation between Candida dubliniensis and Candida albicans by fatty acid methyl ester analysis using gas-liquid chromatography; Peltroche-Llacsahuanga H et al.; Candida dubliniensis is often found in mixed culture with C . albicans, but its recognition is hampered as the color of its colonies in primary culture on CHROMagar Candida varies . Furthermore, definite identification of C . dubliniensis is difficult to achieve, time-consuming, and expensive . Therefore, a method to discriminate between these two closely related yeast species by fatty acid methyl ester (FAME) analysis using gas-liquid chromatography (Sherlock Microbial Identification System {MIS}; MIDI, Inc., Newark, Del.) was developed . Although the chromatograms of these two species revealed no obvious differences when applying FAME analysis, a new library (CADLIB) was successfully created using Sherlock Library Generation Software (MIDI) . The amount and frequency of FAME was analyzed using library training files (n = 10 for each species), preferentially those comprising reference strains . For testing the performance of the CADLIB, clinical isolates genetically assigned to the respective species (C . albicans, n = 32; C . dubliniensis, n = 28) were chromatographically analyzed . For each isolate tested, MIS computed a similarity index (SI) indicating a hierarchy of possible strain fits . When using the newly created library CADLIB, the SIs for C . albicans and C . dubliniensis ranged from 0.11 to 0.96 and 0.53 to 0 . 93 (for all but one), respectively . Only three isolates of C . albicans (9.4%) were misidentified as C . dubliniensis, whereas all isolates of C . dubliniensis were correctly identified . Resulting differentiation accuracy was 90.6% for C . albicans and 100% for C . dubliniensis . Cluster analysis and principal component analysis of the resulting FAME profiles showed two clearly distinguishable clusters matching up with two assigned species for the strains tested . Thus, the created library proved to be well suited to discriminate between these two species. J Clin Microbiol, 2000 Oct, 38(10), 3595 - 607 Elevated phenotypic switching and drug resistance of Candida albicans from human immunodeficiency virus-positive individuals prior to first thrush episode; Vargas K et al.; Strains of Candida albicans obtained from human immunodeficiency virus (HIV)-positive individuals prior to their first episode of oral thrush were already in a high-frequency mode of switching and were far more resistant to a number of antifungal drugs than commensal isolates from healthy individuals . Switching in these isolates also had profound effects both on susceptibility to antifungal drugs and on the levels of secreted proteinase activity . These results suggest that commensal strains colonizing HIV-positive individuals either undergo phenotypic alterations or are replaced prior to the first episode of oral thrush . They also support the suggestion that high-frequency phenotypic switching functions as a higher-order virulence trait, spontaneously generating in colonizing populations variants with alterations in a variety of specific virulence traits. Neurosurgery, 2000 Oct, 47(4), 973 - 6; discussion 976-7 Candida albicans cerebral granulomas associated with a nonfunctional cerebrospinal fluid shunt: case report; Soto-Hernandez JL et al.; OBJECTIVE AND IMPORTANCE: We report an unusual case of basal ganglia granulomas caused by Candida albicans that surrounded the proximal segment of a nonfunctional cerebrospinal fluid shunt in a previously healthy patient . CLINICAL PRESENTATION: A 22-year-old woman had undergone ventriculoatrial cerebrospinal fluid shunt placement for posttraumatic hydrocephalus 3 years previously . One year later, a shunt revision was followed by wound dehiscence with local infection at the neck level . She received oral administration of antibiotics for 3 months until the wound closed . Twelve weeks before admission, the patient experienced pulmonary emboli . She received anticoagulants, and the distal segment of the shunt was removed . Five weeks after shunt removal, she presented with headache and left-sided hemiplegia caused by right basal ganglia inflammatory masses . INTERVENTION: A stereotactic brain biopsy was performed, and the shunt remnants were removed . Microscopically, the lesions were acutely and chronically inflamed . C . albicans grew in tissue and in shunt hardware cultures . The patient was treated with 1.1 g of intravenously administered amphotericin B and orally administered ketoconazole; she recovered completely . CONCLUSION: C . albicans brain granulomas occur rarely in immunocompetent patients . Despite the large size of the lesions and severe brain edema, the absence of an underlying disease contributed to complete resolution after shunt removal and antifungal therapy. Clin Exp Dermatol, 2000 Jul, 25(5), 404 - 5 Persistent annular erythema of infancy associated with intestinal Candida colonization; Stachowitz S et al.; We report a case of persistent annular erythema of infancy in a 4-month-old boy . Physical and laboratory parameters showed no sign of internal disease or specific infection except a massive Candida albicans colonization (> 103 organisms/mm3) of the lower gastrointestinal tract . Oral treatment with amphotericin B for 2 weeks resulted in a complete remission of the skin lesions indicating Candida colonization as a trigger. Braz J Infect Dis, 2000 Apr, 4(2), 47 - 54 Changing strategies for treatment of systemic mycoses; Graybill JR; There have been a number of changes in strategies in antifungal therapy in the past few years . AIDS related mycoses have decreased, and the increase of fluconazole resistant Candida albicans may be slowing because fewer severely immune depressed patients require constant fluconazole suppression . Candida species continue to be relatively common blood culture isolates . About half of these are C . albicans and half non-albicans species . In recent years, we have moved from the use of amphotericin B to fluconazole for initial treatment of candidemia . We have seen fluconazole resistant isolates emerge, primarily C . glabrata and a few C . krusei, but also C . albicans . It is unclear whether the increasing use of fluconazole in intensive care units will worsen this problem . There appears to be no advantage for the lipid formulations of amphotericin B, though they are useful to reduce or prevent renal toxicity . In the United States and Europe, prevention and treatment of aspergillosis have become increasingly important . There are increasing data suggesting that lipid formulations are more effective for both treatment and prevention of invasive disease in the most vulnerable patients with this infection . Renal toxicity is reduced but not avoided by use of the lipid formulations of amphotericin B . For those patients with less acutely progressing disease, the triazoles may be effective options . It is unclear at present whether itraconazole, voriconazole, or posaconazole will be the most favored drug . One promising new class, now in clinical trials, is the echinocandin group . Other agents, such as the sordarins, the chitin synthase inhibitors, and topoisomerase inhibitors, have promise but are much earlier in development . Unfortunately, we still have >50% treatment failure with acute invasive aspergillosis, and 20%-30% failures with candidemia . Now that we have multiple classes of antifungal drugs available, and others in preclinical trials, it would be advantageous to begin more active exploration of combination therapy with antifungals and with combined immune modulators and antifungals. Rev Argent Microbiol, 2000 Jul-Sep, 32(3), 123 - 8 Criteria for Candida albicans numerical analysis based on electrophoretic protein patterns; Boriollo MF et al.; Even in normalized gels, some problems with the choice and position of protein bands always have hindered the processing of electrophoretic data . We have developed a way to establish which criterion best fits the necessities in order to maximize the similarity indexes for numerical analysis . Some repetitions of a Candida albicans strain were carried out in eleven different gels . After staining, the bands were scored in numbers within ranges of +/- n values with increases of one quarter steps (+/- 0.25 kDa, +/- 0.5 kDa, +/- 0.75 kDa, +/- 1.0 kDa, +/- 1.25 kDa) that will limit the possibility of variation for the same bands that will appear on the repetitions . Using this criterion, we have determined that values scored within +/- 1.25 kDa could optimize the minimum limiting value of similarity in different dendrograms built. Biochim Biophys Acta, 2000 Jul 14, 1480(1-2), 117 - 31 Enzymic characteristics of secreted aspartic proteases of Candida albicans; Koelsch G et al.; Candida yeasts are rarely infectious, but frequently cause life-threatening systemic infections in patients immunocompromised by AIDS or by immunosuppressive therapeutics . The secreted aspartic proteases (Saps) are known virulence factors of pernicious Candida species . The most virulent, Candida albicans, possesses at least nine SAP genes, some of which are specifically expressed from cells with morphologies associated with virulence . Only one of these proteases, Sap2, has been previously purified from yeast in sufficient quantities for enzymic studies . The other enzymes are present in low amounts in yeast culture and are difficult to purify . As a consequence, enzyme properties, including the substrate specificities, of all Saps are poorly studied . Therefore, four Saps that are known to be expressed in C . albicans, Sap1, Sap2, Sap3 and Sap6, were produced in Escherichia coli as recombinant zymogens and purified in large quantities . These proenzymes were autoactivated and purified as active proteases . The enzymic properties including the substrate specificities at the P(1) and P(1)' sites were determined using a competitive hydrolysis method employing synthetic substrate mixtures . All four Saps cleave peptide bonds between larger hydrophobic amino acids, but these somewhat broad specificities differ in detail among the four enzymes at both sites . At the P(1) site, Sap1, Sap2 and Sap6 prefer Phe while Sap3 prefers Leu . Positively charged amino acids are also accommodated, especially by Sap2 and Sap3 . The specificities at P(1)' are broader than at P(1) for all four enzymes . Sap6 prefers Ala, whereas other Saps prefer Tyr . Acidic side chains are also accommodated at this site . Analysis of substrates with a hydrophobic amino acid in P(1)' reveals that all the Saps possess a unique preference for Ala at this site . The observed differences of residue preferences among Saps may be utilized for the design of specific substrates and inhibitors. Biochim Biophys Acta, 2000 Jul 24, 1492(2-3), 369 - 76 Functional cloning and mutational analysis of the human cDNA for phosphoacetylglucosamine mutase: identification of the amino acid residues essential for the catalysis; Mio T et al.; In Saccharomyces cerevisiae, phosphoacetylglucosamine mutase is encoded by an essential gene called AGM1 . The