Candida albicans

Gene, 1998 Oct 5, 220(1-2), 91 - 8
A fourth gene from the Candida albicans CDR family of ABC transporters; Franz R et al.; Using primers derived from a region of the Candida albicans CDR1 (Candida drug resistance) gene that is conserved in other ABC (ATP-binding cassette) transporters, a DNA fragment from a previously unknown CDR gene was obtained by polymerase chain reaction (PCR) . After screening a C . albicans genomic library with this fragment as a probe, the complete CDR4 gene was isolated and sequenced . CDR4 codes for a putative ABC transporter of 1490 amino acids with a high degree of homology to Cdr1p, Cdr2p and Cdr3p from C . albicans (62, 59 and 57% amino acid sequence identity, respectively) . Cdr4p has a predicted structure typical for cluster I . 1 of yeast ABC transporters, characterized by two homologous halves, each comprising an N-terminal hydrophilic domain with consensus sequences for ATP binding and a C-terminal hydrophobic domain with six transmembrane helices . In contrast to the CDR1/CDR2 genes, the genetic structure of the CDR4 gene was conserved in 59 C . albicans isolates from six different patients . Northern hybridization analysis showed that the CDR4 gene was expressed in most isolates, but no correlation between CDR4 mRNA levels and the degree of fluconazole resistance of the isolates was found . In addition, a C . albicans mutant in which both copies of the CDR4 gene were disrupted by insertional mutagenesis was not hypersusceptible to fluconazole as compared to the parent strain . Unlike CDR1 and CDR2, CDR4 does not, therefore, seem to be involved in fluconazole resistance in C . albicans.

Res Microbiol, 1998 May, 149(5), 327 - 38
Reaggregation and binding of cell wall proteins from Candida albicans to structural polysaccharides; Aguado C et al.; Urea or hot sodium dodecyl sulphate extracted a significant amount of the same proteins from the matrix of the cell wall of the yeast form and mycelial cells of Candida albicans . Gel filtration analysis of the urea-extracted proteins revealed that they occurred in the form of large complexes which were unaffected by up to 8 M urea . Among them, proteins en route to becoming covalently associated within the wall scaffold were identified by their reaction with specific antibodies . When urea was removed by dialysis, some of these proteins specifically reassociated into large aggregates which bound strongly with ConA, whereas others remained soluble in smaller associated products . The ability of some of these proteins to bind to the insoluble wall polysaccharides was also assessed . No self-assembling proteins were able to bind to glucans and/or chitin . Specificity of the binding to polysaccharides made of beta-bound glucosyl or N-acetylglucosaminyl residues was determined by the competitive effect of several disaccharides . Whereas laminaribiose and diacetylchitobiose were strong inhibitors of protein binding to both glucan and chitin, lactose, maltose and sucrose were ineffective.

Res Microbiol, 1997 Sep-Oct, 148(7), 593 - 603
Initial steps of wall protoplast regeneration in Candida albicans; Rico H et al.; Cell wall regeneration of individual Candida albicans yeast and mycelial protoplasts was studied with confocal and electron microscopy using polyclonal antibodies and lectins . Quantitative measurements of the fluorescence emitted by individual protoplasts during the process of regeneration indicate that chitin is the first polymer to be laid down, whereas beta (1,3)- and beta (1,6)glucan are incorporated at a later stage . Mannoproteins were found on the surface of fresh protoplasts and those newly synthesized were then deposited with time . During the first steps of wall regeneration, the proteins that interacted covalently with chitin or glucan were different, but the same species were found linked to each polymer in yeast and mycelial regenerating forms . The aggregates formed by regenerating protoplasts were shown to be due to the chitin and mannoprotein network initially laid.

J Bacteriol, 1998 Oct, 180(20), 5334 - 43
Identification of Candida albicans ALS2 and ALS4 and localization of als proteins to the fungal cell surface; Hoyer LL et al.; Additional genes in the growing ALS family of Candida albicans were isolated by PCR screening of a genomic fosmid library with primers designed from the consensus tandem-repeat sequence of ALS1 . This procedure yielded fosmids encoding ALS2 and ALS4 . ALS2 and ALS4 conformed to the three-domain structure of ALS genes, which consists of a central domain of tandemly repeated copies of a 108-bp motif, an upstream domain of highly conserved sequences, and a domain of divergent sequences 3' of the tandem repeats . Alignment of five predicted Als protein sequences indicated conservation of N- and C-terminal hydrophobic regions which have the hallmarks of secretory signal sequences and glycosylphosphatidylinositol addition sites, respectively . Heterologous expression of an N-terminal fragment of Als1p in Saccharomyces cerevisiae demonstrated function of the putative signal sequence with cleavage following Ala17 . This signal sequence cleavage site was conserved in the four other Als proteins analyzed, suggesting identical processing of each protein . Primary-structure features of the five Als proteins suggested a cell-surface localization, which was confirmed by indirect immunofluorescence with an anti-Als antiserum . Staining was observed on mother yeasts and germ tubes, although the intensity of staining on the mother yeast decreased with elongation of the germ tube . Similar to other ALS genes, ALS2 and ALS4 were differentially regulated . ALS4 expression was correlated with the growth phase of the culture; ALS2 expression was not observed under many different in vitro growth conditions . The data presented here demonstrate that ALS genes encode cell-surface proteins and support the conclusion that the size and number of Als proteins on the C . albicans cell surface vary with strain and growth conditions.

Int J STD AIDS, 1998 Sep, 9(9), 526 - 30
Vulvovaginal candidiasis in female sex workers; Otero L et al.; Vulvovaginal candidiasis is a frequent inflammatory process in women but it has not been widely studied in female sex workers (FSWs) . To estimate the frequency of Candida species infection in FSWs and to identify related risk factors and clinical findings, we carried out a retrospective study of 1923 FSWs over 11 years . We also performed a prospective study of 163 consecutive FSWs with a history of candidiasis during a 4-year period . Candida species were isolated in 1967 samples (18.5% of the total) . Candida albicans (89.3%) was the most frequent species, followed by Candida glabrata (2.7%), Candida parapsilosis (1.2%) and Saccharomyces cerevisiae (0.4%) . In the prospective study of 163 patients, we found vaginal discharge in 76.1% of cases, soreness in 52.1% and vulval pruritus in 32.5% . We identified 12 patients (7.4%) with recurrent vulvovaginal candidiasis . No statistical difference was found between recurrent vulvovaginitis and the use of oral contraceptives, oral sex, tight-fitting clothing and synthetic underwear . FSWs have the same prevalence of candidiasis as other groups of women described in published literature . The proportion of albicans and non-albicans species does not differ between women with recurrent and non-recurrent vulvovaginal candidiasis (VVC).

AIDS, 1998 Sep 10, 12(13), 1601 - 10
Change in fluconazole susceptibility patterns and genetic relationship among oral Candida albicans isolates; Diaz-Guerra TM et al.; OBJECTIVE: To assess the genetic homogeneity or heterogeneity within each set of Candida albicans isolates colonizing/infecting the oral cavities of HIV-infected patients undergoing azole therapy when changes in susceptibility to fluconazole were detected . DESIGN: Fourteen HIV-positive patients suffering recurrent episodes of oral candidosis were prospectively followed from the first episode to the isolation of strains with decreased susceptibility to fluconazole . The strains of C . albicans isolated either from episodes or controls throughout the prospective study were analysed . METHODS: Electrophoretic karyotyping and hybridization with the repeated sequence probe 27A were used to delineate sequential isolates . In vitro susceptibility tests to fluconazole and ketoconazole were also performed . The results obtained by DNA fingerprinting with the probe combined with computer-assisted analysis were used to assess the genetic relationships amongst the strains . In addition, comparison with the genetic relatedness of a group of geographically unrelated strains was made . RESULTS: Isogenic populations of sequential isolates were observed only in two patients; 12 patients harboured heterogenic populations over time, although in 11 patients there was a predominant strain that was isolated more than once, and only one of these patients carried strains with a similarity index less than 80% . With the exception of two patients, each patient carried a major strain that became less susceptible to fluconazole . The similarity index for the unrelated strains was 59% . CONCLUSIONS: HIV-infected patients may carry a mixed population of strains, but the strains tend to be related to each other . The strains were maintained throughout the course of infection and at least one developed secondary resistance to fluconazole.

Farmaco, 1998 Jun 30, 53(6), 415 - 20
Synthesis of some new benzimidazolecarboxamides and evaluation of their antimicrobial activity; Goker H et al.; A series of 1,2-disubstituted benzimidazole-5(6)-carboxamides was prepared and evaluated in vitro for antimicrobial activity against Staphyloccus aureus, Escherichia coli and Candida albicans . The precursor benzimidazolecarboxylic acids 4a-c and 9a-c were prepared via oxidative condensation of diaminobenzoic acids with aldehydes and via several steps over the 2(1H)-benzimidazolones, respectively . All acids were converted to their acyl chlorides with SOCl2, then amidified with several N,N'-dialkylaminoethyl derivatives . Compounds 8a-c, 20 and 22 exhibited the best activity.

Exp Gerontol, 1998 Aug, 33(5), 477 - 84
Age-associated differences in neutrophil oxidative burst (chemiluminescence); Braga PC et al.; Phagocytic defensive functions consist of a sequence of events, including migration, phagocytosis, secretion, and the release of reactive oxygen species (ROS) . The last of these (also called "oxidative burst") has not received due attention in the elderly, even though it can be considered the most important event in the process of killing an invading microorganism . The aim of the present study was to investigate the oxidative burst activity of polymorphonuclear neutrophil leukocytes (PMNs) in relation to age, using a technique that specifically identifies ROS production: luminol-amplified chemiluminescence (LACL) . Besides the use of LACL, a particular feature of the study was the use of five rather than just one or two different stimulants: two particulate (Candida albicans and zymosan) and three soluble ones {N-formyl-methionyl-leucyl-phenylalanine (fMLP), phorbol 12 myristate 13 acetate (PMA), and polyanetholesulfonate (liquoid)} . This approach allowed us to observe a dichotomy between the effects of Candida and zymosan (particulates), which were not significantly different in the elderly subjects compared to the young controls, and those of fMLP, PMA, and liquoid (solubles), which showed a significant reduction in LACL in the elderly group . Considering the different results obtained with the various stimulants adopted that are all believed to have NADPH oxidase as a common final target of oxidative burst, it may be postulated that aging can influence the different transductional pathways in different ways.

Otolaryngol Pol, 1998, 52(3), 277 - 80
{The role of imperfect fungi in etiopathogenesis of allergic rhinitis}; Namyslowski G et al.; In this paper the role of imperfect fungi in etiopathogenesis of perennial rhinitis was examined . In a group of 26 patients the concentration of total IgE and IgE specific of Candida albicans, Aspergillus fumigatus, Alternaria alternata, Mucor racemosus and Cladosporum herbarum was signified . Oversensibility to imperfect fungi was confirmed in 30.8% of patients: to Candida albicans in 3.8%, to Aspergillus fumigatus in 11.5%, to Alternaria alternata in 3.8%, to Mucor racemosus in 7.6% and to Cladosporum herbarum in 3.8%.

Biochemistry, 1998 Oct 6, 37(40), 14326 - 36
Binding and reactivity of Candida albicans estrogen binding protein with steroid and other substrates; Buckman J et al.; In this report recombinant estrogen binding protein (EBP1), isolated originally from Candida albicans as a result of its high affinity for 17beta-estradiol, has been purified extensively using a modified affinity purification scheme originally developed for a homolog of EBP1, old yellow enzyme (OYE) . It is shown that like OYE, the protein binds a variety of compounds with a phenolic structure, including 17beta-estradiol, and compounds with an alpha, beta-unsaturated keto or aldehyde structure . In addition, EBP1 exhibits an NADPH oxidoreductase activity, transferring electrons from NADPH to all alpha,beta-unsaturated ketones and aldehydes tested via the tightly bound FMN cofactor . Analysis of the steady-state kinetics of these reactions indicate a tetra uni ping-pong mechanism . Inhibition of the steady-state reaction by 17beta-estradiol gives a Ki = 10 +/- 2 nM, and indicates exclusive binding of this steroid to the enzyme in its oxidized state . In contrast, 19-nortestosterone binds to both oxidized and reduced forms of the enzyme with dissociation constants of 600 +/- 100 and 650 +/- 90 nM, respectively . EBP1 also catalyzes a disproportionation reaction with certain compounds, in which two molecules of a cylic alpha,beta-unsaturated ketone, including the steroid 19-nortestosterone, are individually aromatized and reduced to the corresponding saturated ketone . Despite the extensive similarity in sequence and enzymic activity, notable differences between EBP1 and the OYE family of proteins exist with regard to the binding behavior and reactivity with the two steroids tested here, estradiol and 19-nortestosterone.

J Immunol, 1998 Oct 1, 161(7), 3543 - 50
IFN-gamma is required for IL-12 responsiveness in mice with Candida albicans infection; Cenci E et al.; To elucidate the role of IFN-gamma in antifungal CD4+ Th-dependent immunity, 129/Sv/Ev mice deficient for IFN-gamma receptor (IFN-gammaR(-/-)) were assessed for susceptibility to gastrointestinal or systemic Candida albicans infection and for parameters of innate and adaptive T helper immunity . IFN-gammaR(-/-) mice failed to mount protective Th1-mediated acquired immunity upon mucosal immunization or in response to a live vaccine strain of the yeast . The impaired Th1-mediated resistance correlated with defective IL-12 responsiveness, but not IL-12 production, and occurred in the presence of an increased innate antifungal resistance . The development of nonprotective Th2 responses was observed in IFN-gammaR(-/-) mice upon mucosal infection and subsequent reinfection . However, under experimental conditions of Th2 cell activation, the occurrence of Th2 cell responses was similar in IFN-gammaR(-/-) and in IFN-gammaR(+/+) mice . These results indicate the complex immunoregulatory role of IFN-gamma in the induction of mucosal and nonmucosal anticandidal Th cell responses; IFN-gamma is not essential for the occurrence of Th2 responses but is required for development of IL-12-dependent protective Th1-dependent immunity.

Ann Allergy Asthma Immunol, 1998 Sep, 81(3), 247 - 55
IgE-sensitization to cellular and culture filtrates of fungal extracts in patients with atopic dermatitis; Nissen D et al.; BACKGROUND: Patients with atopic dermatitis may experience exacerbations of eczema triggered by various inflammatory stimuli . One mechanism may be IgE-mediated reactions to dermatophytes since these patients are more likely to acquire skin infections with dermatophytes and may become sensitized . OBJECTIVE: This study investigates IgE-sensitization to fungi in patients with atopic dermatitis and compares the biologic activity of culture filtrates and cellular fungal extracts . The following allergen extracts were provided as culture filtrates and cellular extracts: Candida albicans, Fusarium moniliforme, and Penicillium notatum . In addition, Pityrosporum ovale and Trichophyton rubrum cultures were included in the test panel . METHODS: Fifteen patients with clinical findings suggesting dermatophytosis and 11 controls were selected . Each subject was tested by leukocyte histamine release and skin prick test to each fungal extract . The extracts were separated and reduced by sodium dodecylsulfate polyacrylamide gel electrophoresis and analyzed by IgE-immunoblotting with sera from all study subjects . RESULTS: Fourteen patients (93%) reacted to one or several fungal extracts by releasing histamine when challenged in vitro . By immunoblotting experiments, patient sera showed binding to a wide range of components in all extracts . Patient sera recognized allergenic components shared by culture filtrates and cellular extracts but with higher frequent and greater intensity in culture filtrates . Although culture filtrates generated more frequent and potent IgE-reactions than the cellular extracts, the difference was not statistically significant . Biologic potency was similar when evaluated by skin prick tests and leukocyte histamine release . CONCLUSION: Patients with atopic dermatitis may develop specific IgE-antibodies to a number of fungi as demonstrated by IgE-immunoblotting . In selected patients, fungi may trigger an IgE-mediated reaction that may contribute to the exacerbation of eczema . Approximately, one-half of the patients, however, produced IgE-antibodies to fungal (glyco)proteins without a significant histamine release or skin test response possibly because of nonspecific interaction with carbohydrate moieties on IgE and poor biologic activity of IgE antibodies directed to cross-reactive carbohydrate determinants of fungal glycoproteins . This warrants caution when interpreting clinical relevance of serologic measurements of fungal IgE-antibodies.

Antimicrob Agents Chemother, 1998 Oct, 42(10), 2745 - 6
Enhancement of antimicrobial activity of neuropeptide Y by N-terminal truncation; Shimizu M et al.; The activity of neuropeptide Y (NPY) against Candida albicans, which was revealed to be fungicidal, was enhanced significantly by the truncation of amino acid residues at the N terminus . The most active peptides (MICs, approximately 1 microM) were about 10-fold more potent than the intact NPY (MIC, approximately 10 microM) . The enhancement was weakened by the replacement of the N terminus by negatively charged residues and/or acylation of the alpha-amino group . These results suggest that only the alpha-helical region of NPY is necessary for the antimicrobial activity and that the net charge of the peptide is important for the activity.

Antimicrob Agents Chemother, 1998 Oct, 42(10), 2645 - 9
Mechanism of fluconazole resistance in Candida krusei; Orozco AS et al.; The mechanisms of fluconazole resistance in three clinical isolates of Candida krusei were investigated . Analysis of sterols of organisms grown in the absence and presence of fluconazole demonstrated that the predominant sterol of C . krusei is ergosterol and that fluconazole inhibits 14alpha-demethylase in this organism . The 14alpha-demethylase activity in cell extracts of C . krusei was 16- to 46-fold more resistant to inhibition by fluconazole than was 14alpha-demethylase activity in cell extracts of two fluconazole-susceptible strains of Candida albicans . Comparing the carbon monoxide difference spectra of microsomes from C . krusei with those of microsomes from C . albicans indicated that the total cytochrome P-450 content of C . krusei is similar to that of C . albicans . The Soret absorption maximum in these spectra was located at 448 nm for C . krusei and at 450 nm for C . albicans . Finally, the fluconazole accumulation of two of the C . krusei isolates was similar to if not greater than that of C . albicans . Thus, there are significant qualitative differences between the 14alpha-demethylase of C . albicans and C . krusei . In addition, fluconazole resistance in these strains of C . krusei appears to be mediated predominantly by a reduced susceptibility of 14alpha-demethylase to inhibition by this drug.

Antimicrob Agents Chemother, 1998 Oct, 42(10), 2584 - 9
Rapid, transient fluconazole resistance in Candida albicans is associated with increased mRNA levels of CDR; Marr KA et al.; Fluconazole-resistant Candida albicans, a cause of recurrent oropharyngeal candidiasis in patients with human immunodeficiency virus infection, has recently emerged as a cause of candidiasis in patients receiving cancer chemotherapy and marrow transplantation (MT) . In this study, we performed detailed molecular analyses of a series of C . albicans isolates from an MT patient who developed disseminated candidiasis caused by an azole-resistant strain 2 weeks after initiation of fluconazole prophylaxis (K . A . Marr, T . C . White, J . A . H . vanBurik, and R . A . Bowden, Clin . Infect . Dis . 25:908-910, 1997) . DNA sequence analysis of the gene (ERG11) for the azole target enzyme, lanosterol demethylase, revealed no difference between sensitive and resistant isolates . A sterol biosynthesis assay revealed no difference in sterol intermediates between the sensitive and resistant isolates . Northern blotting, performed to quantify mRNA levels of genes encoding enzymes in the ergosterol biosynthesis pathway (ERG7, ERG9, and ERG11) and genes encoding efflux pumps (MDR1, ABC1, YCF, and CDR), revealed that azole resistance in this series is associated with increased mRNA levels for members of the ATP binding cassette (ABC) transporter superfamily, CDR genes . Serial growth of resistant isolates in azole-free media resulted in an increased susceptibility to azole drugs and corresponding decreased mRNA levels for the CDR genes . These results suggest that C . albicans can become transiently resistant to azole drugs rapidly after exposure to fluconazole, in association with increased expression of ABC transporter efflux pumps.

Antimicrob Agents Chemother, 1998 Oct, 42(10), 2534 - 41
Identification and characterization of novel antimicrobial decapeptides generated by combinatorial chemistry; Hong SY et al.; Novel combinatorial libraries consisting of simplified amino acid sequences were designed to screen for peptides active against the Candida albicans membrane . A novel decapeptide, KKVVFKVKFK, that had a unique primary amino acid sequence was identified in this work . This peptide irreversibly inhibited the growth of C . albicans and showed a broad range of antibacterial activity but no hemolytic activity . Circular dichroism spectra revealed that the predominant secondary structure of this peptide strongly depended on the membrane-mimetic environments; the peptide preferred to form an amphipathic alpha-helical structure in the presence of 50% trifluoroethanol, while it preferred to adopt a distorted alpha-helical structure in the presence of sodium dodecyl sulfate micelles . Experiments in which dye was released from vesicles indicated that this novel antimicrobial peptide killed microorganisms through the action on the membrane as its primary target . Replacement of amino acids in this active decapeptide on the basis of information from the libraries could provide unique information about factors affecting its antimicrobial activity such as its secondary structure, net positive charge, and hydrophobicity.

Chemotherapy, 1998 Nov-Dec, 44(6), 405 - 8
Effects of cefepime, cefixime and ceftibuten on murine gut colonization by Candida albicans; Maraki S et al.; Crl:CD1(ICR) BR mice were fed chow containing Candida albicans or regular chow . Both groups were subsequently given either antibiotics or normal saline . Stool cultures were performed before, at the end of treatment and 1 week after treatment, to determine the effect on the stool yeast concentration . Candida-colonized mice treated with cefepime, cefixime or ceftibuten had higher (however not significantly) counts of the yeast in their stools than control Candida-fed mice treated with saline . A group of Candida-fed mice were treated with ceftriaxone, which is known to increase the yeast stool concentration significantly and served as positive control . Mice fed regular chow and treated with the study drugs or saline did not have any yeasts in their stools . Dissemination of Candida did not occur.

FEBS Lett, 1998 Sep 11, 435(1), 49 - 54
Isolation and characterization of the Candida albicans gene for mRNA 5'-triphosphatase: association of mRNA 5'-triphosphatase and mRNA 5'-guanylyltransferase activities is essential for the function of mRNA 5'-capping enzyme in vivo; Yamada-Okabe T et al.; The amino acid sequence of the Saccharomyces cerevisiae mRNA 5'-triphosphatase (TPase) diverges from those of higher eukaryotes . In order to confirm the sequence divergence of TPases in lower and higher eukaryotes, the Candida albicans gene for TPase was identified and characterized . This gene designated CaCET1 (C . albicans mRNA 5'-capping enzyme triphosphatase 1) has an open reading frame of 1.5 kb, which can encode a 59-kDa protein . Although the N-terminal one-fifth of S . cerevisiae TPase (ScCet1p) is missing in CaCet1p, CaCet1p shares significant sequence similarity with ScCet1p over the entire region of the protein; the recombinant CaCet1p, which was expressed as a fusion protein with glutathione S-transferase (GST), displayed TPase activity in vitro . CaCET1 rescued CET1-deficient S . cerevisiae cells when expressed under the control of the ADH1 promoter, whereas the human capping enzyme derivatives that are active for TPase activity but defective in mRNA 5'-guanylyltransferase (GTase) activity did not . Yeast two-hybrid analysis revealed that C . albicans Cet1p can bind to the S . cerevisiae GTase in addition to its own partner, the C . albicans GTase . In contrast, neither the full-length human capping enzyme nor its TPase domain interacted with the yeast GTase . These results indicate that the failure of the human TPase activity to complement an S . cerevisiae cet1delta null mutation is attributable, at least in part, to the inability of the human capping enzyme to associate with the yeast GTase, and that the physical association of GTase and TPase is essential for the function of the capping enzyme in vivo.

Mycopathologia, 1998, 141(3), 137 - 42
Monoclonal antibodies directed against extracellular matrix proteins reduce the adherence of Candida albicans to HEp-2 cells; Cotter G et al.; The presence of the extracellular matrix (ECM) proteins collagen types I and IV, laminin and fibronectin on the surface of HEp-2 cells was confirmed by flow cytometry using monoclonal antibodies . Monoclonal antibodies directed against these ECM proteins reduced the adherence of C . albicans ATCC 44990 to HEp-2 cells, the greatest reductions being evident in assays which incorporated anti-collagen type IV monoclonal antibody . The ability of sugaramines to inhibit the adherence of C . albicans to a variety of cell types has been demonstrated previously and the most significant reduction in C . albicans-HEp-2 adherence was in assays which incorporated 0.2M galactosamine . The combination of anti-collagen IV monoclonal antibody and galactosamine reduced the adherence of C . albicans to HEp-2 cells by approximately 70% (p < 0.05).

Mycopathologia, 1998, 141(3), 127 - 35
Isolation of a petite mutant from a histidine auxotroph of Candida albicans and its characterization; Roth-Ben Arie Z et al.; Respiration-deficient (petite) mutations have been induced in various yeasts, which are categorized as petite-positive . Candida albicans was classified among the petite-negative yeasts . Since then, a few reports have appeared, describing the isolation of petite mutants in C . albicans . We report in the present study on the isolation of a petite mutant of C . albicans-SAR1 . This mutant was isolated from a histidine auxotroph of C . albicans after mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine, thus our petite mutant carries a double mutation . SAR1 was characterized morphologically, biochemically and ultrastructurally . The results revealed differences from the wild type in respect to morphological, physiological and biochemical characteristics . Electron microscopy showed that the cells of the petite mutant contain only very few mitochondria that looked 'thread like' without any cristae . The significance of the mutation in the virulence of the mutant vs . that of the wild-type is being assessed.

J Dent, 1998 Sep, 26(7), 577 - 83
Adherence of Candida albicans to denture-base materials with different surface finishes; Radford DR et al.; OBJECTIVES: To assess the in vitro adherence of Candida albicans to heat-cured hard and soft denture-base materials with varying surface roughness, and to observe the effect of a mixed salivary pellicle on candidal adhesion to these surfaces . METHODS: In vitro adhesion assays on heat-cured acrylic resin (Trevalon), Molloplast B and Novus using the type strain of C . albicans (NCPF 3153A) . Surfaces for the assays were prepared using clinically appropriate rotary instruments . Unstimulated, pooled and clarified whole saliva was used to assess its effect on adhesion . RESULTS: Significantly greater adhesion of C . albicans to rough rather than smooth surfaces was found (P < 0.001), as well as increased adhesion to the machined soft lining materials compared with acrylic . Pre-coating denture-base materials with saliva reduced candidal adhesion on all materials . CONCLUSIONS: Rough surfaces on denture-base materials promote the adhesion of C . albicans in vitro . However, saliva reduces adhesion of C . albicans and thus diminishes the effect of surface roughness and free surface energy differences between materials.

Gene, 1998 Sep 18, 218(1-2), 85 - 93
The CARE-2 and rel-2 repetitive elements of Candida albicans contain LTR fragments of a new retrotransposon; Goodwin TJ et al.; CARE-2 and Rel-2 are dispersed, repetitive elements of Candida albicans . Hybridisation experiments suggest that they are present at 10-20 copies per genome and appear on most, if not all, of the chromosomes . A high degree of interstrain variation has been demonstrated for CARE-2, making it of use for strain typing . Until now, however, the nature of the repetitive elements within CARE-2 and Rel-2 was unknown . We show here that CARE-2 and Rel-2 contain long terminal repeat (LTR) fragments of a new retrotransposon . These LTRs, which we designate kappa, are partially responsible for the repetitive nature of CARE-2 and Rel-2 . Complete copies of the kappa elements are present elsewhere in the genome and adjacent to some are sequences characteristic of the internal regions of retrotransposons . An apparently high degree of scrambling of the kappa elements suggests that they may represent a hotspot for mutation and recombination in C . albicans.

Mycopathologia, 1998, 141(2), 105 - 9
Activity of hydrolytic enzymes of Candida albicans strains isolated from patients with periodontal and membrane mucosae of oral cavity diseases; Kurnatowska AJ; Fungi are elements of the ontocenosis of the oral cavity and causal factors of inflammatory lesions in its mucous membrane . The objective of the study was to find differences in the activity of hydrolytic enzymes of Candida albicans isolated from patients with diseases of the periodontium and mucous membrane of the oral cavity . Of 235 patients examined, 31 were diagnosed with gingivitis, 38 with glossitis, 28 with leucoplakia, 37 with adult periodontitis, 25 with juvenile periodontitis, 36 stomatitis prothetica and 40 with stomatitis atrophica . In 196 patients (83.4 +/- 2.4%), fungi belonging to Candida species were detected . In the evaluation of Candida albicans strains (146) properties, bioMerieux API ZYM tests containing substrates for the detection of 19 hydrolases were used . All the investigated strains were characterized by the activity of 14 enzymes, i.e . phosphatase alcaline, esterase (C4), esterase lipase (C8), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, alpha galactosidase, beta galactosidase, alpha glucosidase, beta glucosidase, N-acetyl-beta-glucosaminidase, alpha mannosidase and alpha fucosidase . Strains isolated from the oral cavity of patients with disease of periodontium and mucous membrane are characterised by the highest phosphatase acid activity . The greatest enzymatic activity is characteristic of Candida albicans isolated from patients with stomatitis atrophica or stomatitis prothetica, and the lowest in strains from gingivitis or juvenile periodontitis cases . Differences in the activity of hydrolases are statistically significant (p < 0.01) for: esterase (C4), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, beta glucosidase, N-acetyl-beta-glucosaminidase, of fungi isolated from patients with particular clinical diagnoses.

Mycopathologia, 1998, 141(2), 59 - 63
Regulation of superoxide dismutase synthesis in Candida albicans; Gunasekaran U et al.; The synthesis of superoxide dismutase {SOD: EC 1.15.1.1} in response to various cultural conditions was examined in Candida albicans, an opportunistic yeast which causes candidiasis in immunosuppressed patients . SOD plays an important role in protecting cells from teh oxidative damage of superoxide radicals . Maximum SOD activity was found after 72 hrs of yeast growth . The optimum pH and temperature for the SOD activity were 7 and 40 degrees C, respectively . The major SOD activity was found in the cytosol fraction and the level of extracellular SOD was very low . The enzyme was stimulated to varying degrees by cholic acid, procaine and tocopherol . On the basis of inhibitor studies and other enzyme properties, the isolated enzyme from C . albicans is identified as a copper and zinc superoxide dismutase.

Epidemiol Mikrobiol Imunol, 1998 Aug, 47(3), 87 - 92
{Personal experience with classification of yeast microorganisms . I . The combined biotyping method of Mencl and Otcenásek and typing using the "killer" phenomenon}; Hamal P et al.; Using a group of 150 isolates of Candida albicans, C . glabrata, C . krusei, C . tropicalis, C . parapsilosis a C . kefyr the differentiating capacity of two biotyping systems was tested-the combined method according to Mencl and Otcenasek, and typing using the so-called killer phenomenon . With the combined method comparable results with the original work of the authors were obtained . This applies to the number of biotypes as well as to the ratio of the dominant biotype . As regards the differentiating characteristics of different biotypes the two studies differed fundamentally . As to typing, using the "killer" phenomenon, its practical usefulness was tested, the differentiating capacity of the method was, however, very much influenced by the small number of available killer-positive yeast strains.

Ceska Slov Farm, 1998 Jul, 47(4), 186 - 8
{Antimicrobial activity of (N-salicylidene-DL-aspartate- and (N-salicylidene-L-asparaginate)-copper complexes with pyrazole-type ligands}; Sokolik J et al.; By a reaction of salicylaldehyde (Scl) with the corresponding amino acids and by the next complexation reaction of the formed Schiff bases with Cu2+ ions in an aqueous-alcoholic medium, aqua (N-salicylideneaminoalkanoato)copper(II) complex chelates of the composition Cu(Scl-DL-Asp(2-)) (H2O)2, Ip and Cu(Scl-L-Asn(2-)(H2O), In were prepared . The monodiazole complexes with pyrazole IIp and IIn (as monohydrate) as well as with 3,5-dimethylpyrazole IIIp a IIIn were prepared by replacing the molecule of H2O in the parent aquacomplexes with the diazoles under the same reaction conditions . Using a routine dilution micromethod, the antimicrobial activity of the prepared complexes and free diazoles was tested against Staphylococcus aureus, Escherichia coli and Candida albicans . Only a significant antistaphylococcus activity was found (highest for the complex IIn; MIC = 39 micrograms/cm3) . All chelates (Ip,n-IIIp,n) were more effective (MIC = 39-156 micrograms/cm3) than both pyrazole (312 micrograms/cm3)and 3,5-dimethylpyrazole (625 micrograms/cm3) alone . The relationship between the coordination-chemical properties and the biological effects of the complexes studied is discussed.

Infect Immun, 1998 Oct, 66(10), 4845 - 50
Mannan-specific immunoglobulin G antibodies in normal human serum accelerate binding of C3 to Candida albicans via the alternative complement pathway; Zhang MX et al.; Candida albicans activates the classical and alternative complement pathways, leading to deposition of opsonic complement fragments on the cell surface . Our previous studies found that antimannan immunoglobulin G (IgG) in normal human serum (NHS) allows C . albicans to initiate the classical pathway . The purpose of this study was to determine whether antimannan IgG also plays a role in initiation of the alternative pathway . Pooled NHS was rendered free of classical pathway activity by chelation of serum Ca2+ with EGTA alone or in combination with immunoaffinity removal of antimannan antibodies . Kinetic analysis revealed a 6-min lag in detection of C3 binding to C . albicans incubated in EGTA-chelated NHS, compared to a 12-min lag in NHS that was both EGTA chelated and mannan absorbed . The 12-min lag was shortened to 6 min by addition of affinity-purified antimannan IgG . The accelerating effect of antimannan IgG on alternative pathway initiation was dose dependent and was reproduced in a complement binding reaction consisting of six purified proteins of the alternative pathway . Both Fab and F(ab')2 fragments of antimannan IgG facilitated alternative pathway initiation in a manner similar to that observed with intact antibody . Immunofluorescence analysis showed that addition of antimannan IgG to EGTA-chelated and mannan-absorbed serum promoted an early deposition of C3 molecules on the yeast cells but had little or no effect on distribution of the cellular sites for C3 activation . Thus, antimannan IgG antibodies play an important regulatory role in interactions between the host complement system and C . albicans.

Curr Genet, 1998 Sep, 34(3), 192 - 9
Identification of polymorphic mutant alleles of CaMDR1, a major facilitator of Candida albicans which confers multidrug resistance, and its in vitro transcriptional activation; Gupta V et al.; CaMDR1 (Candida albicans Multi Drug Resistance) encodes a major facilitator whose expression in Saccharomyces cerevisiae confers resistance to several unrelated drugs . We describe here the identification and molecular characterization of seven mutant alleles of CaMDR1 (CaMDR1-1 to 1-7) . The complete sequencing of CaMDR1 alleles revealed several in-frame point mutations leading to a change in amino-acid residues where insertion/replacement of an aspartate residue in a serine-asparagine-aspartate-rich domain was most noteworthy . Interestingly, these alleles showed a distinct drug resistance profile . The expression of CaMDR1, or of its alleles, in C . albicans cells was enhanced by benomyl, methotrexate and several other unrelated drugs, and was more pronounced in at least one of the azole-resistant clinical isolates.

Rev Argent Microbiol, 1998 Apr-Jun, 30(2), 100 - 3
{Rapid differentiation and presumptive identificaiton of yeasts using Candida CHROM-agar medium}; Giusiano GE et al.; In order to evaluate faster and cheaper methods for isolation and identification of clinically important yeasts, we used CHROM-agar Candida (CAC), a chromogenic medium, for differentiation and presumptive identification . A total of 546 yeast strains were studied . Strains were previously identified by conventional methods . The colour and texture of Candida albicans, C . tropicalis, C . krusei and Trichosporon beigelii were always constant and particularly distinctive for a reliable identification . C . glabrata, Saccharomyces cerevisiae, C . parapsilopsis colonies also showed constant colour . Because of its contents of cloramphenicol, CAC is also suitable for isolation and observation of mixed yeast species from clinical samples . CAC is a useful medium for reliable differentiation and presumptive identification of clinically important yeasts, particularly form immunocompromised patients.

J Immunol, 1998 Sep 15, 161(6), 2684 - 91
HindIII liposomes suppress delayed-type hypersensitivity responses in vivo and induce epidermal IL-10 in vitro; Nishigori C et al.; Considerable evidence suggests that ultraviolet-B (UV-B) radiation suppresses certain immune responses through the induction of cyclobutane pyrimidine dimers in DNA . To determine whether induction of other forms of DNA damage in the skin mimicked the immunosuppressive effects of UV-B radiation, we produced double-strand breaks in the DNA of epidermal cells with HindIII restriction endonuclease encapsulated in liposomes . Application of these liposomes, but not liposomes containing inactive HindIII or an irrelevant endonuclease, to the skin of C3H mice suppressed the induction of delayed-type hypersensitivity responses to Candida albicans and alloantigen and induced IL-10 production in the epidermis . Treatment of the Pam212 murine keratinocyte cell line with these liposomes in vitro induced immunosuppressive activity and IL-10 in culture supernatants . Unlike UV-B irradiation, however, HindIII in liposomes failed to induce suppressor T cell activity in vivo or in vitro . We conclude that double-strand breaks in DNA of epidermal cells can induce immunosuppression and up-regulate the production of immunomodulatory cytokines; however, either DNA damage alone does not account for all the immunosuppressive properties of UV-B irradiation, or cyclobutane pyrimidine dimers differ qualitatively from double-strand breaks in their biologic consequences . These studies raise the possibility that drugs causing DNA damage may induce cytokine dysregulation and immune suppression in addition to cytotoxicity.

FEMS Microbiol Lett, 1998 Sep 1, 166(1), 135 - 9
Mitogen-activated protein kinase-defective Candida albicans is avirulent in a novel model of localized murine candidiasis; Guhad FA et al.; Candida albicans strains with a deletion of the mitogen-activated protein kinase CEK1 gene are defective in the yeast to hyphal transition on solid surfaces in vitro . The virulence of a cek1 delta/cek1 delta null mutant strain was compared with its wild-type parent strain (WT) in a novel model of localized candidiasis . The mammary glands of lactating mice (at day 5 postpartum) were infected for 2, 4 and 6 days with 50 microliter suspension containing 1 x 10(5), 1 x 10(6) and 1 x 10(7) blastopores before death . Infected and non-infected control glands were evaluated pathologically . All animals infected with cek1 delta/cek1 delta null mutant strains showed no lesions while 65% of animals infected with the WT strain had severe lesions characterized by widespread heterophilic infiltration, necrosis, and abscess formation . As an additional control, animals infected with the disrupted strain complemented with the WT CEK1, on a replicating plasmid, also showed severe pathological changes similar to the WT strain . These results clearly demonstrate that the CEK1 gene codes for a virulence determinant of C . albicans and that the mouse mastitis model is well suited for the discriminative study of the pathogenicity of different C . albicans strains.

Ann Dermatol Venereol, 1997, 124(2), 157 - 8
{Pustular candidiasis in heroin addicts}; Leclech C et al.; INTRODUCTION: Pustular candidiasis in heroin addicts is a rare entity in dermatology . We report a case . CASE REPORT: A 29-year-old female heroin addict developed a painful pustular growth on the scalp . There was no fever . Multiple follicular pustulae measuring 2 to 3 mm were associated with hyperesthesia of the scalp and painful cervical nodes . Biopsy showed acute ostiofolliculitis with a few blastospores and mycelial filaments . Candida albicans was isolated from the pustulae and the buccal cavity . Candida serology was positive (indirect immunofluorescence 1/100, coelectrosyneresis: 4 archs) . Search for other localizations and HIV serology were negative . The last injection of brown heroin had been taken 15 days earlier; lemon had been added . Treatment with flucanazole (400 mg/d) led to improvement within 48 hours . DISCUSSION: Sudden development of pustulae or nodules in pilous zones in a heroin addict should suggest the diagnosis . Outcome depends on early treatment after diagnosis and search for other localizations . Our case presented two particular aspects: ostiofollicular localization of the pustulae and a long delay (15 days) between the (presumably) last injection and the development of the lesion . Folliculitis develops almost exclusively in addicts who use brown heroin . Contamination by Candida albicans results from the lemon used to improve solubility at injection.

APMIS, 1998 Jul, 106(7), 736 - 42
Adhesion to denture acrylic surfaces and relative cell-surface hydrophobicity of Candida parapsilosis and Candida albicans; Panagoda GJ et al.; C . parapsilosis is an opportunistic emerging pathogen which together with C . albicans causes diseases in immunocompromised patients . Adhesion of Candida species to various surfaces is an important event in colonization and pathogenesis, and the relative cell-surface hydrophobicity (CSH) of the organism is a contributory physical force involved . Therefore, in vitro adhesion to acrylic surfaces and relative CSH of 24 isolates of C . parapsilosis and 10 isolates of C . albicans were studied . There was no significant difference in relative adhesion of C . parapsilosis isolates and C . albicans, although the former demonstrated a tendency for increased adhesion . There was significant intra-species variation in adhesion among isolates of C . parapsilosis (p=0.0001), but not C . albicans . In general, C . parapsilosis isolates demonstrated a two-fold greater relative CSH than C . albicans (p=0.0003) . When the relative CSH of superficial and systemic isolates of C . parapsilosis were compared, the former showed a significantly higher (49.15%) relative CSH than their systemic counterparts (p<0.01) . A highly significant positive correlation between adhesion and relative CSH of C . parapsilosis (p=0.74, p<0.0001) was also noted . Taken together, these data suggest that the attributes of adhesion and relative CSH of Candida species may contribute differentially in varying disease states of the human host, such as superficial and systemic Candida infections.

J Clin Microbiol, 1998 Oct, 36(10), 3007 - 12
Detection of Candida dubliniensis in oropharyngeal samples from human immunodeficiency virus-infected patients in North America by primary CHROMagar candida screening and susceptibility testing of isolates; Kirkpatrick WR et al.; Candida dubliniensis has been associated with oropharyngeal candidiasis in patients infected with human immunodeficiency virus (HIV) . C . dubliniensis isolates may have been improperly characterized as atypical Candida albicans due to the phenotypic similarity between the two species . Prospective screening of oral rinses from 63 HIV-infected patients detected atypical dark green isolates on CHROMagar Candida compared to typical C . albicans isolates, which are light green . Forty-eight atypical isolates and three control strains were characterized by germ tube formation, differential growth at 37, 42, and 45 degreesC, identification by API 20C, fluorescence, chlamydoconidium production, and fingerprinting by Ca3 probe DNA hybridization patterns . All isolates were germ tube positive . Very poor or no growth occurred at 42 degreesC with 22 of 51 isolates . All 22 poorly growing isolates at 42 degreesC and one isolate with growth at 42 degreesC showed weak hybridization of the Ca3 probe with genomic DNA, consistent with C . dubliniensis identification . No C . dubliniensis isolate but only 18 of 28 C . albicans isolates grew at 45 degreesC . Other phenotypic or morphologic tests were less reliable in differentiating C . dubliniensis from C . albicans . Antifungal susceptibility testing showed fluconazole MICs ranging from </=0.125 to 64 microgram/ml . Two isolates were resistant to fluconazole (MIC, 64 microgram/ml) and one strain was dose dependent susceptible (MIC, 16 microgram/ml) . MICs of other azoles, including voriconazole, itraconazole, and SCH 56592, for these isolates were lower . C . dubliniensis was identified in 11 of 63 (17%) serially evaluated patients . Variability in phenotypic characteristics dictates the use of molecular and biochemical techniques to identify C . dubliniensis . This study identifies C . dubliniensis in HIV-infected patients from San Antonio, Tex., and shows that C . dubliniensis is frequently detected in those patients by using a primary CHROMagar screen.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2440 - 2
Microbicidal activity of a new silver-containing polymer, SPI-ARGENT II; Kampf G et al.; The survival of three bacterial species and Candida albicans was studied on SPI-ARGENT II . The immediate recovery from silver-impregnated polymer and control polymer (1 cm2) was approximately 10(6) to 10(7) microorganisms . After incubation (37 degreesC) and neutralization of silver with horse serum (5%), surviving organisms were recovered . The survival of the microorganisms on the polymer was not found to be influenced by the silver implantation.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2446 - 8
Susceptibilities of Candida species isolated from the lower gastrointestinal tracts of high-risk patients to the new semisynthetic echinocandin LY303366 and other antifungal agents; Zhanel GG et al.; Fifty-two percent of stool specimens collected from 1,200 high-risk patients were colonized with yeasts, primarily Candida albicans (53 . 6%) and Candida glabrata (35.7%) . Susceptibilities to all antifungal agents tested, including LY303366, were similar to those reported previously for Candida species isolated from blood.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2437 - 9
Activity of liposomal amphotericin B with prolonged circulation in blood versus those of AmBisome and fungizone against intracellular Candida albicans in murine peritoneal macrophages; van Etten EW et al.; Activity against intracellular Candida albicans was assessed in C . albicans-infected murine peritoneal macrophages exposed to long-circulating pegylated amphotericin B liposomes (PEG-AMB-LIP), AmBisome, or Fungizone . The level of antifungal activity of Fungizone is much higher than that of AmBisome or PEG-AMB-LIP, while PEG-AMB-LIP and AmBisome show equivalent activity levels . Previous exposure of uninfected macrophages to PEG-AMB-LIP or AmBisome is advantageous for intracellular antifungal activity.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2434 - 6
Control of Candida albicans murine vaginitis by topical administration of polycarbophil-econazole complex; Ghelardi E et al.; The complexation of econazole with the mucoadhesive polycarbophil was found to significantly improve the therapeutic benefit of the drug in the topical treatment of experimental vaginal candidiasis in mice, while no difference in the antimycotic activity exerted by econazole and polycarbophil-econazole could be detected in vitro.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2431 - 3
Superior efficacy of liposomal amphotericin B with prolonged circulation in blood in the treatment of severe candidiasis in leukopenic mice; van Etten EW et al.; In leukopenic mice with severe systemic candidiasis, single-dose treatment (5 mg of amphotericin B {AMB}/kg of body weight) with long-circulating polyethylene glycol-coated AMB liposomes (PEG-AMB-LIP) resulted in zero mortality and a significant reduction in the number of viable Candida albicans in the kidney, whereas 70% mortality was seen in mice treated with five daily doses of AmBisome (5 mg of AMB/kg . day) . When the first of five daily doses of AmBisome was combined with a single low dose of Fungizone (0.1 mg of AMB/kg), the efficacy was equal to that of PEG-AMB-LIP.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2405 - 9
Pharmacokinetics of pentoxifylline and its metabolites in healthy mice and in mice infected with Candida albicans; Miller K et al.; Pentoxifylline has immunomodulatory properties and has been shown to decrease organ damage and improve survival in animals with gram-negative sepsis or endotoxemia . This effect is mediated by a reduction in endotoxin-induced production of tumor necrosis factor alpha (TNF-alpha) by the host . In earlier studies, we observed an unexpected increase in mortality in mice infected with Candida albicans that were given pentoxifylline even though concentrations of TNF-alpha in serum were not affected . The current study was designed to determine whether the pharmacokinetics of pentoxifylline and its metabolites were altered in C . albicans-infected mice and, if so, whether these changes could have contributed to the increased mortality . Noninfected mice and mice infected with C . albicans were treated with pentoxifylline (60 mg/kg of body weight) intraperitoneally every 8 h . Serum was collected from animals after one (day 0), four (day 1), or seven (day 2) injections of pentoxifylline or saline (controls) . The first dose was administered 6 h after C . albicans infection . Serum was pooled . Concentrations of pentoxifylline and metabolites I, IV, and V were determined by capillary gas chromatography . Renal function and hepatic profiles were assessed . Pharmacokinetic parameters (maximum concentration of pentoxifylline in serum, half-life, and area under the concentration-time curve from 0 h to infinity {AUC(0)-infinity}) for all noninfected mice were similar and did not differ from those for day 0-infected mice . For day 1-infected mice, values of these three pharmacokinetic parameters for pentoxifylline and metabolite I were increased two- to fourfold over values for noninfected and day 0-infected mice . For metabolites IV and V, the AUC(0)-infinity was increased approximately eightfold over control values . In addition, day 1-infected mice demonstrated evidence of renal and hepatic dysfunction . In summary, C . albicans infection produced marked changes in the pharmacokinetics of pentoxifylline and its metabolites in the mice . The high concentrations of pentoxifylline and its metabolites in serum attained in infected mice may have contributed to the increased mortality of mice with systemic candidiasis.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2342 - 6
Synergy of nitric oxide and azoles against Candida species in vitro; McElhaney-Feser GE et al.; The candidacidal activity of nitric oxide (NO) as delivered by a class of compounds termed diazeniumdiolates has been investigated . Diazeniumdiolates are stable agents capable of releasing NO in a biologically usable form at a predicted rate, and three such compounds were examined for activity . One compound, (Z)-1-{N-(2-aminoethyl)-N-(2-ammonioethyl)amino}diazen-1- ium-1, 2-diolate (DETA-NO), proved to be most suitable for examining NO activity due to its relatively long half-life (20 h) and because of limited candidacidal activity of the uncomplexed DETA nucleophile . DETA-NO was active against six species of Candida for which the MICs necessary to inhibit 50% growth (MIC50s) ranged from 0.25 to 1.0 mg/ml . C . parapsilosis and C . krusei were the most susceptible to the compound . In addition to a determination of NO effects alone, the complex was utilized to investigate the synergistic potential of released NO in combination with ketoconazole, fluconazole, and miconazole . Activity was investigated in vitro against representative strains of Candida albicans, C . krusei, C . parapsilosis, C . tropicalis, C . glabrata, and C . dubliniensis . Determination of MIC50, MIC80 and MICs indicated that DETA-NO inhibits all strains tested, with strains of C . parapsilosis and C . krusei being consistently the most sensitive . The combination of DETA-NO with each azole was synergistic against all strains tested as measured by fractional inhibitory concentration indices that ranged from 0.1222 to 0.4583 . The data suggest that DETA-NO or compounds with similar properties may be useful in the development of new therapeutic strategies for treatment of Candida infections.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2279 - 83
Identification of elongation factor 2 as the essential protein targeted by sordarins in Candida albicans; Dominguez JM et al.; The target for sordarins in Candida albicans has been elucidated . Kinetic experiments of sordarin inhibition as well as displacement experiments showed that the formation of a sordarin-target complex follows a reversible mechanism . Binding of tritiated drug to the target is enhanced in the presence of ribosomes . Isolation of the target by classical protein purification methods has allowed us to identify it as elongation factor 2 . This is in agreement with the nature of sordarin derivatives as specific inhibitors of the elongation cycle within protein synthesis in yeasts.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2197 - 205
Molecular mode of action of the antifungal beta-amino acid BAY 10-8888; Ziegelbauer K et al.; BAY 10-8888 is a cyclic beta-amino acid that is related to cispentacin and that has antifungal activity . Candida albicans cells accumulated BAY 10-8888 intracellularly to a concentration about 200 that in the medium when grown in media with a variety of nitrogen sources . In complex growth medium, BAY 10-8888 transport activity was markedly reduced and was paralleled by a decrease in its antifungal activity . Uptake of BAY 10-8888 was mediated by an H+-coupled amino acid transporter with specificity for branched-chain amino acids (isoleucine, leucine, and valine) and showed a KT (Michaelis constant of the transport reaction) of 0.95 mM and a Vmax of 18.9 nmol x min-1 x 10(7) cells-1 . Similar to the transport of natural amino acids in Saccharomyces cerevisiae, the transport of BAY 10-8888 into the cell was unidirectional . Efflux occurred by diffusion and was not carrier mediated . Inside the cell BAY 10-8888 inhibited specifically isoleucyl-tRNA synthetase, resulting in inhibition of protein synthesis and cell growth . Intracellular isoleucine reversed BAY 10-8888-induced growth inhibition . BAY 10-8888 was not incorporated into proteins . BAY 10-8888 inhibited isoleucyl-tRNA synthetase with the same concentration dependency as protein biosynthesis in intact cells assuming 200-fold accumulation.

Immunopharmacol Immunotoxicol, 1998 Aug, 20(3), 421 - 31
Protective effect of oral administration of a traditional medicine, Juzen-Taiho-To, and its components on lethal Candida albicans infection in immunosuppressed mice; Abe S et al.; Protective effects of a kampo medicine, Juzen-taiho-to (TJ-48) and its herbal components against experimental candidiasis in cyclophosphamide-induced immunosuppressive mice were investigated . ICR mice were immunosuppressed by intraperitoneal treatment with cyclophosphamide (day-4) and were orally given TJ-48 or one of its 10 herbal components for 4 consecutive days (day-4--1) . They were then challenged intravenously with a lethal dose of Candida albicans (day 0) . An oral dose of 1 g/kg/day of TJ-48 prolonged their life span . A similar protective effect was obtained by treatment with its component drugs Ginseng radix, Glycyrrhizae radix, Atractylodis lancea rhizoma or Cnidii rhizoma . These herbal components were suggested to have a main role in the protective effect of Juzen-taiho-to against Candida infection.

Bol Asoc Med P R, 1998 Jan-Mar, 90(1-3), 21 - 6
Anti-fungal and cytokine producing activities of CD8 + T lymphocytes from HIV-1 infected individuals; Colon MD et al.; Lymphokine activated killer (LAK) cells are capable of killing not only malignant cells but also hyphal form of Candida albicans in vitro . When peripheral blood mononuclear cells (PBMC) from normal healthy donors were cultured for 72-96 hrs with 1,500 international unit (IU)/ml interleukin-2 (IL-2), marked LAK activity was induced . However, even prior to IL-2 activation, PBMC isolated from some normal subjects and those from almost all individuals who are infected by human immunodeficiency virus type 1 (HIV-1) exhibited significant levels of anti-fungal activity . Such pre-activation ("in situ") antifungal activity of PBMC decreased during the initial 48 hrs of IL-2 activation . PBMC from HIV-1 seropositive subjects showed higher levels of "in situ" anti-fungal activity than normal PBMC did . After a decline of "in situ" activity during the initial 48 hours, LAK activity gradually increased and reached near maximal levels by day 4 and remained more or less constant until day 6 . No significant difference was observed between the LAK activity of normal and HIV-1(+) PBMCs on days 4-6 . In IL-2 activated normal and HIV-1(+) PBMC cultures, both CD4 and CD8 T cells produced IL-2, INF-gamma as well as TNF-alpha . Production of IL-2 by both CD4 and CD8 T cells was suppressed in HIV-1(+) PBMC cultures, but no significant suppression of INF-gamma production was noted . Meanwhile, TNF-alpha production by CD4 was very much suppressed but no significant changes in TNF-alpha production by CD8 T cells was noted in HIV-1(+) PBMC cultures.

Scand J Infect Dis, 1998, 30(2), 137 - 42
Outbreak of Candida albicans fungaemia in a neonatal intensive care unit; Huang YC et al.; During a 4-month period, 9 premature infants hospitalized in a neonatal intensive care unit (NICU) developed Candida albicans fungaemia . All 9 infants received antifungal agents . Fluconazole was administered in 7 patients and successfully eradicated this organism in 6 with no adverse effects . For epidemiological investigation, 64 environmental specimens and hand-washings of all 54 staff members involved in the NICU were examined for the presence of this organism . No C . albicans could be identified from environmental sources, while the hand-washing of 1 nurse was C . albicans-positive . Two genotyping methods, including electrophoretic karyotyping using contour-clamped homogeneous electric field gel electrophoresis and polymerase chain reaction-based direct sequencing of rRNA gene, were used in the analysis of the isolates recovered from blood cultures of the infants, the hand-washing of the nurse and 7 control isolates . Both methods yielded comparable results and revealed that all 13 isolates from infected infants as well as the isolate from hand washing of the nurse were of the same genotype while the control isolates were distinct . These results suggest that the outbreak of C . albicans fungaemia was caused by a particular strain and possibly via cross-infection . In addition, we showed that fluconazole seemed to be safe and effective in treating C . albicans fungaemia in neonates, although the data were limited.

J Infect Dis, 1998 Sep, 178(3), 792 - 802
Early signal transduction induced by Candida albicans in macrophages through shedding of a glycolipid; Jouault T et al.; Cell wall beta-1,2-oligomannosides are involved in Candida albicans binding to macrophages and in their stimulation to produce cytokines . The nature of signaling events occurring during initial interaction of macrophage J774 cell line and C . albicans, together with the nature of molecules containing beta-1,2-oligomannosides released by the yeasts, was examined . Cocultivation led to a herbimycin A-sensitive production of tumor necrosis factor-alpha . Immunofluorescence and Western blotting confirmed tyrosine phosphorylation and revealed an accumulation of 90- to 120-kDa phosphoproteins . Antibodies specific for beta-1,2-oligomannosides showed that these epitopes were shed at an early stage from the yeasts to the macrophage membrane, in association with a glycolipid previously described as C . albicans phospholipomannan . Incubation of macrophages with purified phospholipomannan alone led to a signal transduction pathway identical to that observed with living yeasts . All of these results demonstrate that C . albicans phospholipomannan shedding is involved in C . albicans-macrophage interaction through beta-1,2-oligomannosides.

Eur J Pediatr, 1998 Aug, 157(8), 661 - 2
Pharmacokinetics of oral fluconazole in premature infants; Wenzl TG et al.; Systemic infections with Candida albicans in neonates are a frequent and well recognized problem . The therapeutic gold standard in this situation is the combined intravenous antimycotic treatment with amphotericin B and flucytosine . Potential adverse effects of this regimen have encouraged the search for desirable alternatives . We report on the use of oral fluconazole in neonates with Candida albicans septicaemia . Three premature infants were treated with four courses of therapy . Pharmacokinetic studies were performed during each course . At oral doses of 4.5-6 mg/kg once a day, serum levels of fluconazole were within the therapeutic range during the entire dosage interval . Follow up showed microbiological and clinical cure in all patients with no side-effects . In one patient a dosage of 4 mg/kg per day lead to a microbiological relapse with sub-therapeutic serum levels . CONCLUSIONS: Oral fluconazole seems to be a safe and effective treatment for Candida albicans septicaemia even in premature infants.

Eur Respir J, 1998 Aug, 12(2), 502 - 4
Eosinophilic lung disease associated with Candida albicans allergy; Pacheco A et al.; A significant number of cases of chronic eosinophilic pneumonia remain idiopathic in spite of a comprehensive search of associated causes . This study reports a patient with a classical clinical presentation of chronic eosinophilic pneumonia and peripheral blood eosinophilia in whom selective sensitization to Candida albicans was demonstrated . This yeast was present in the bronchoalveolar lavage culture and specific serum immunoglobulin (Ig)E and IgG against C . albicans were found in the patient's serum . Levels of these specific immunoglobulins diminished with corticosteroid treatment and increased coinciding with a new outbreak of symptoms after lowering the dosage of corticosteroids . To the author's knowledge, this is the first case described of chronic eosinophilic pneumonia associated with sensitization to C . albicans . Evaluation of allergy to C . albicans should be performed in chronic eosinophilic pneumonia before labelling cases as idiopathic.

J Antibiot (Tokyo), 1998 Jul, 51(7), 665 - 75
Characterization and optimization of in vitro assay conditions for (1,3)beta-glucan synthase activity from Aspergillus fumigatus and Candida albicans for enzyme inhibition screening; Wood RL et al.; (1,3)Beta-D-glucan synthase (E.C.2.4.1.34 . UDP-glucose: 1,3-beta-D-glucan 3-beta-glucosyl transferase) catalyzes the polymerization of glucose ({1-3}-beta-linkages) using UDP-glucose as substrate . We have determined optimal in vitro conditions for the assay of (1,3)beta-glucan synthase activity from Aspergillus fumigatus and Candida albicans . These included lysis of cells in the following for C . albicans, 100 mM HEPES, pH 8.0, 10 microM guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS), 2 mM ethylenediaminetetraacetic acid (EDTA), disodium salt, 5 mM NaF, 250 mM sucrose, and 10 mM NaH2PO4; and for A . fumigatus, 50 mM HEPES, 10mM EDTA, 750 mM sucrose, 10 mM NaH2PO4, 100 mM cellobiose and 50 microM GTPgammaS . Resulting low-speed supernatants were used as enzyme sources to determine the optimal in vitro assay conditions . We have characterized the resulting enzyme activities and tested the optimized assays with known (1,3)beta-glucan synthase inhibitors including cilofungin, papulacandin, aculeacin A, and echinocandin B . We have used both optimized assays to screen > 1000 extracts of marine macroorganisms and, using bioassay-guided purification, have identified (1,3)beta-glucan synthase inhibitors.

Mycopathologia, 1998, 141(1), 1 - 6
Factors influencing the expression in vitro of Candida albicans stress mannoproteins reactive with salivary secretory IgA; Vidotto V et al.; We have examined the influence of subinhibitory concentrations of several antifungals, the different glucose and ammonium sulphate concentrations in the culture medium as well as the strain variability on the expression in vitro of stress mannoproteins reactive with salivary sIgA in C . albicans and other Candida spp isolates . Irrespective of the conditions used, no reactivity with salivary sIgA was observed in yeast cells grown at 25 degrees C . However, when grown at 37 degrees C, all of the 10 C . albicans strains, but only 9 out of 28 non-C . albicans isolates studied showed reactivity with salivary sIgA . Cells grown at 37 degrees C in medium containing maximum concentrations of glucose and ammonium sulphate expressed the antigens reactive with sIgA during longer periods of time than the cells grown in medium with minimal concentrations of the same compounds . The regulatory role showed by the concentration of glucose and ammonium sulphate on the antigenic expression was subordinated, nevertheless, to the most important factor, the temperature of incubation . Only isolates showing low susceptibility expressed the antigens reactive with sIgA under the influence of subinhibitory concentration of antifungals . However, induced resistance to one of the antifungals tested (5 fluorocytosine) allowed the antigenic expression at elevated subinhibitory concentrations even in previous susceptible strains . In conclusion, in addition to the temperature, factors such as characteristics of the strain, the concentration of glucose and ammonium sulphate in the culture medium and the resistance to antifungals played a role on the expression of C . albicans antigens reactive with sIgA, which could be of clinical relevance in the course of infection.

Mol Microbiol, 1998 Aug, 29(3), 753 - 62
Deletion analysis of yeast Sec65p reveals a central domain that is sufficient for function in vivo; Regnacq M et al.; The Saccharomyces cerevisiae SEC65 gene encodes a 32 kDa subunit of yeast signal recognition particle that is homologous to human SRP19 . Sequence comparisons suggest that the yeast protein comprises three distinct domains . The central domain (residues 98-171) exhibits substantial sequence similarity to the 144 residue SRP19 . In contrast, the N-terminal and C-terminal domains (residues 1-97 and 172-273 respectively) share no similarity to SRP19, with the exception of a cluster of positively charged residues at the extreme C-terminus of both proteins . Here, we report the cloning of a Sec65p homologue from the yeast Candida albicans that shares the same extended domain structure as its S . cerevisiae counterpart . This conservation of sequence is reflected at the functional level, as the C . albicans gene can complement the conditional lethal sec65-1 mutation in S . cerevisiae . In order to examine the role of the N- and C- terminal domains in Sec65p function, we have engineered truncation mutants of S . cerevisiae SEC65 and tested these for complementing activity in vivo and for SRP integrity in vitro . These studies indicate that a minimal Sec65p comprising residues 76-209, which includes the entire central SRP19-like domain, is sufficient for SRP function in yeast.

Ann Allergy Asthma Immunol, 1998 Aug, 81(2), 165 - 9
Recurrent vaginal candidiasis and allergic rhinitis: a common association; Moraes PS; BACKGROUND: In a recent series of studies, it has been shown that hypersensitivity is an important factor in recurrent vaginal candidiasis and several referring gynecologists have been asking for our support in the evaluation of women with regard to the possibility of local vaginal hypersensitivity . This study was instituted because, since the first anamnesis, a high incidence of perennial allergic rhinitis, as well as family allergies have been observed in these patients . OBJECTIVE: To study the association between recurrent vaginal candidiasis and perennial allergic rhinitis, and to explore the allergic characteristics of the patients and of the disease . METHODS: For 28 months, we have prospectively studied 95 patients with recurrent vaginal candidiasis referred by gynecologists to a private allergy practice . All of them were unresponsive to all other modalities of therapy, and had no diabetes or immunodeficiency diseases . As a control group, we studied 100 women, who came to the allergy office for other reasons, and had no recurrent vaginal candidiasis . All of these 195 women were submitted to a standard allergy medical history, a complete physical examination, and immediate skin tests with a standard battery of inhalant allergens and Candida albicans . The incidence of allergic diseases in the two groups was compared . RESULTS: Sixty-four patients with recurrent vaginal candidiasis also had allergic rhinitis (71%), while in the control group the incidence of this pathology was 42% . The difference was considered statistically significant, according to the software Epi info (P < .0001) . The study (1) did not show an association between recurrent vaginal candidiasis and asthma and (2) indicated that patients with recurrent vaginal candidiasis have a high incidence of skin tests positive to inhalant allergens (50%), to Candida albicans (55%), and a high incidence of family history of allergies (73%) . In control women, the incidence of skin test positive to inhalant allergens was 72% and to Candida albicans, 10% . There was a family history of allergies in 61% of women . CONCLUSIONS: The data demonstrate that recurrent vaginal candidiasis is statistically associated with perennial allergic rhinitis and that many of these women with recurrent vaginal candidiasis tend to be atopic.

J Biol Chem, 1998 Sep 4, 273(36), 23376 - 80
A functional soluble form of the murine mannose receptor is produced by macrophages in vitro and is present in mouse serum; Martinez-Pomares L et al.; A soluble form of the mannose receptor (sMR) has been found in conditioned medium of primary macrophages in vitro and in mouse serum . sMR was released as a single species, had a smaller size than the cell-associated form, and accumulated in macrophage-conditioned medium, in a cytokine-regulated manner, to levels comparable with those found for cell-associated mannose receptor . Pulse-chase experiments showed that sMR production in culture occurred by constitutive cleavage of pre-existing full-length protein . A binding assay was developed to determine the sugar specificity of sMR and its ability to interact with pathogens and particulate antigens (i.e . Candida albicans and zymosan) . Protease inhibitor studies suggested that sMR was produced by cleavage of an intact mannose receptor by a matrix metalloprotease or ADAM metalloprotease . A role for sMR in the immune response is proposed based on its binding properties, regulation by cytokines, and the previous discovery of putative ligands for the cysteine-rich domain of the mannose receptor in lymph nodes and spleen.

Mol Microbiol, 1998 Jul, 29(2), 605 - 15
Differential expression of secreted aspartyl proteinases in a model of human oral candidosis and in patient samples from the oral cavity; Schaller M et al.; Candida albicans, an opportunistic pathogen in humans, secretes secretory aspartyl proteinases (Saps), which have been correlated with virulence . We examined the temporal regulation of the mRNA expression of seven known members of the SAP gene family by reverse transcription polymerase chain reaction (RT-PCR) in (i) an in vitro model of oral candidosis based on reconstituted human epithelium (RHE); and (ii) clinical samples from patients with oral candidosis . SAP1 and SAP3 transcripts were first detected 42 h after inoculation of RHE, while at the same time, slight morphological alterations in the epithelium were documented by light microscopy . SAP6 expression occurred 6 h later concomitantly with germ tube formation of some infecting Candida cells and severe lesions of the epithelial tissue . SAP2 and SAP8 RT-PCR products were first detected 60 h after infection, while SAP4 and SAP5 transcripts were never discovered . Thus, a temporal progression of SAP expression in the order SAP1 and SAP3 > SAP6 > SAP2 and SAP8 was observed at the same time as increasing RHE damage occurred . At the protein level, Sap antigen was found within the C . albicans yeast cells and the epithelial cells by immunoelectron microscopy using an anti-Sap murine monoclonal antibody directed against the gene products Sap1-3 . Expression of SAP1-3 and 6 was also detected by RT-PCR in samples from patients suffering from oral candidosis . Our results suggest that the pathogenesis of experimental and clinical oral candidosis is associated with the differential and temporal regulation of SAP gene expression.

Clin Exp Allergy, 1998 Jul, 28(7), 808 - 16
Requirement of CD28-CD86 costimulation for allergen-specific T cell proliferation and cytokine expression; Van Neerven RJ et al.; BACKGROUND: Allergen-specific T lymphocytes biased to the production of type 2 cytokines play an important role in the pathophysiology of atopic disease . It is not known whether optimal activation of these T cells requires costimulation via interaction of B7 (CD86/CD80) with CD28 . METHODS: Peripheral blood mononuclear cells (PBMC), isolated from 10 house dust mite Dermatophagoides pteronyssinus (Der p)-allergic asthma patients and 10 non-allergic control individuals, were stimulated with house dust mite (Der p) and the control antigens Candida albicans (CA) and tetanus toxoid (TT) . The role of costimulation in activation for proliferation and cytokine mRNA production of peripheral blood T cells was studied by blocking CD28, CTLA-4, and their ligands CD80 (B7-1) and CD86 (B7-2) . RESULTS: The proliferation and the production of type 1 and 2 cytokine mRNA by T cells in response to Der p as well as the control antigens TT and CA was inhibited by simultaneously masking CD80 and CD86 using CTLA4-Ig, a soluble form of CTLA-4 . Notably, Der p-specific proliferation of T cells from Der p-allergic asthma patients and non-allergic controls were inhibited equally well . Additional experiments with MoAbs revealed that activation of these T cells was optimally inhibited by blocking the interaction of CD28 with CD86 . CONCLUSION: In vitro responses of allergen- and antigen-specific T cells of allergic patients and non-allergic control persons are equally dependent on costimulation via the CD28-CD86 pathway, suggesting that inhibition of this pathway may prevent complete activation of allergen-specific T cells in allergic individuals in vivo.

Microbiology, 1998 Aug, 144 ( Pt 8), 2311 - 21
Disruption studies of a Candida albicans gene, ELF1: a member of the ATP-binding cassette family; Sturtevant J et al.; A 3.6 kb gene (ELF1) with homology to the ATP-binding cassette (ABC) gene family has been isolated from genomic libraries of Candida albicans . Members of this gene family include both membrane transport proteins which confer a drug-resistance phenotype, and proteins whose functions are associated with protein translation . ELF1 (Elongation Like Factor) showed greatest homology with a Saccharomyces cerevisiae ORF (YPL226W), whose function is unknown, and lower homology with fungal elongation factor 3 (EF-3) genes . In comparison, homology with a gene conferring a drug-resistant phenotype (CDR1) was low . To understand the function of ELF1 in C . albicans, gene-knockout experiments were conducted using the hisG-URA3-hisG disruption cassette . Both single-copy (heterozygote) and double-disrupted strains in ELF1 were isolated . Phenotypically, the disrupted strains grew more slowly than wild-type and produced a mixture of large, irregular cells and apparently normal cells.

Microbiology, 1998 Aug, 144 ( Pt 8), 2299 - 310
Overexpression of Candida albicans secretory aspartyl proteinase 2 and its expression in Saccharomyces cerevisiae do not augment virulence in mice; Dubois N et al.; To elucidate the implications of secreted aspartyl proteinase (Sap)2p in the pathogenesis of Candida infections, the SAP2 gene was expressed in Saccharomyces cerevisiae and overexpressed in Candida albicans . The coding region of SAP2, including its signal sequence and propeptide, was amplified by PCR and cloned downstream of the S . cerevisiae or C . albicans ADH1 promoter . Plasmid expression of SAP2 in S . cerevisiae showed that the signal peptide was functional . Integrative transformation of S . cerevisiae and C . albicans was accomplished by homologous recombination within the URA3 locus for S . cerevisiae and the SAP2 locus for C . albicans . Negative control transformants carried plasmids either without the SAP2 insert or with mutated sap2 . S . cerevisiae and C . albicans transformants showed similar growth rates to their parental strains or negative controls, when grown in medium containing amino acids . However, in medium with BSA as sole nitrogen source, constitutive expression of SAP2 enabled S . cerevisiae to grow and increased the growth rate of C . albicans . In both media, only S . cerevisiae transformants harbouring SAP2 secreted the enzyme, as confirmed by proteinase activity assays and immunoblotting . When C . albicans was grown in amino acids medium, the enzyme was detected exclusively in transformants constitutively expressing SAP2 . However, in BSA medium these strains secreted enzyme earlier and secreted higher amounts of enzyme and total proteinase activity . In pathogenicity studies in intact mice, expression of Sap2p as a sole putative virulence factor did not cause S . cerevisiae to become virulent and constitutive overexpression of SAP2 did not augment virulence of C . albicans in experimental oral or systemic infection.

Microbiology, 1998 Aug, 144 ( Pt 8), 2291 - 8
Evaluation of the intranasal challenge route in mice as a mucosal model for Candida albicans infection; Londono P et al.; The intranasal route was used to study Candida albicans infections in mice . Mice from two different inbred strains were challenged intranasally with C . albicans and the level of local and systemic colonization was monitored . DBA/2 mice were highly susceptible to challenge and viable C . albicans disseminated from the lungs to deeper tissues, including kidneys, liver and spleen within 48 h . In contrast, in BALB/c mice challenged in the same manner, C . albicans were retained within the lungs and cleared . Local and systemic anti-C . albicans immune responses were investigated . BALB/c mice exhibited higher titres of serum and mucosal anti-C . albicans IgA than DBA/2 mice . Splenocytes from BALB/c mice, but not from DBA/2 mice, produced detectable levels of interleukin-4 and -5 following stimulation with C . albicans antigens . Both DBA/2- and BALB/c-derived splenocytes produced interferon-gamma and interleukin-10 in response to similar stimulation . In conclusion, the intranasal route provided a simple, non-invasive murine model for investigating C . albicans infection through mucosal surfaces.

FEMS Immunol Med Microbiol, 1998 Jul, 21(3), 223 - 30
Soluble mannan and beta-glucan inhibit the uptake of Malassezia furfur by human monocytic cell line, THP-1; Suzuki T et al.; The uptake of live and heat-killed Malassezia furfur HIC 3321, HIC 3343 and Candida albicans ATCC 10231 by human monocytic cell line, THP-1, was examined . THP-1 was differentiated by PMA for 7 days before use . The uptake of these yeasts by THP-1 was increased in a concentration-dependent manner of yeasts, and the uptake reached plateau level at the E/T (yeast/THP-1) ratio 5 . In addition, a higher percentage of heat-killed cells than live cells was taken in THP-1 . Yeast mannan and beta-1,3-glucan, random coiled conformer, inhibited the uptake of live and heat-killed M . furfur by THP-1, though dextran T-250, that is alpha-glucan, and schizophyllan (SPG), triple helix conformer of beta-glucan, did not . Interestingly, mannan inhibited the uptake of both types, live and heat-killed, of C . albicans, however, laminaran inhibited the uptake of heat-killed C . albicans alone . Opsonization of these yeasts with normal human serum enhanced the uptake of yeasts, although opsonization with heat-inactivated serum, the treatment at 56 degrees C for 30 min, did not enhance . These results suggested that live and heat-killed M . furfur was recognized by THP-1 through mannose receptor, beta-glucan receptor and complement receptor type 3 via the activation of alternative pathway of complement.

Mycoses, 1998, 41 Suppl 1, 71 - 7
{Differentiation and characterization of yeasts pathogenic for humans (Candida albicans, Exophiala dermatitidis) and algae pathogenic for animals (Prototheca spp.) using Fourier transform infrared spectroscopy (FTIR) in comparison with conventional methods}; Schmalreck AF et al.; Due to the Fourier-Transform Infrared Spectroscopy (FT-IR) of strain specific traits demonstrated to be a suitable and efficient method for diagnostic and epidemiological determinations for the yeasts Candida albicans, Exophiala dermatitidis and the chlorophylless algae of the genus Prototheca . FT-IR leads in a rapid and economical way to reproducible results according to the spectral differences of intact cells (IR-fingerprints) . Different genera, species and sub-species respectively, different strains can be recognized and grouped into different clusters and subclusters . The FT-IR analysis of Candida albicans isolates (n = 150) of 22 newborns-at-risk of an intensive care unit showed, that 86% of the children were colonised with several (2-4) different strains in the oral cavities and faeces . Stationary cross-infections could definitely be determined . Exophiala dermatitidis isolates (n = 31), mostly isolated repetitively within a period of 3 years from sputa of patients suffering from cystic fibrosis could be characterized and grouped patient-specifically over the total sampling period . Of 6 from 8 patients (75%) their individual strains remain the same and could be tracked over the three years . Cross-infections during the stationary treatment could be clearly identified by FT-IR . The Prototheca isolate (n = 43) from live-stock and farm environment showed clear distinguishable clusters differentiating the species P . wickerhamii, P . zopfii and P . stagnora . In addition, the biotypes of P . zopfii could be distinguished, especially the subclusters of variants II and III . It could be demonstrated, that FT-IR is suitable for the routine identification and differentiation of yeasts and algae . However, in spite of the gain of knowledge by using FT-IR for the characterization of microorganisms, the conventional phenotyping and/or genetic analysis of yeast or algae strains cannot be replaced completely . For a final taxonomic classification a combination of conventional methods on FT-IR together with more sophisticated molecular genetic procedures is necessary.

Mycoses, 1998, 41 Suppl 1, 47 - 50
{What functions do six different genes for secretory proteinases have in Candida albicans?}; Hube B et al.; Secreted Aspartate Proteinases (Sap) are among those factors of the human pathogen Candida albicans, which promote infections in the immunocompromised host . Sap isoenzymes are encoded by at least nine different genes (SAP1-9), which are differentially regulated in vitro . RT-PCR analysis during experimental infections and from patient samples confirmed the expression of SAP genes in vivo . However, while Sap2 is the dominant isoenzyme under culture conditions, other SAP genes are also expressed during infections . In order to investigate the role of single isoenzymes during the pathogenesis of candidosis, mutants were produced which harbour deletions in SAP1, SAP2, SAP3 and SAP4-6 . Although only SAP2 and SAP4-6 mutants showed a strong reduction of proteolytic activity in vitro, all SAP mutants were significantly attenuated in systemic infections . In addition, SAP2, SAP3 and SAP4-6 mutants were clearly more sensitive to neutrophilic leucocytes compared to the wild type SC5314 . These investigations show that several proteinase isoenzymes are likely to be involved in the pathogenesis of candidosis.

Mycoses, 1998, 41 Suppl 1, 39 - 46
{Qualitative and quantitative studies of autofluorescence in fungi}; Graf B et al.; Fluorescence microscopy is an important method in mycology . It is a common procedure used in immunology or histology and more recently in modern techniques of molecular biology like in-situ hybridization . Since several molds and yeasts show autofluorescence, an interference of this phenomenon with the detection method cannot be excluded . Therefore, we studied autofluorescence in fungi in more detail, in particular with respect to the dependence of this phenomenon from growth conditions, fixing method or mounting medium used . Here we show that moulds cultivated in a liquid medium are strongly autofluorescent which could be considerably reduced by repetitive washing . In moulds, we did not find important differences in autofluorescence levels with the three fixing methods under study . However, this finding cannot be generalized . Thus, in the yeast Candida albicans we found the autofluorescence pattern being largely dependent from the fixing method and the excitation wave length, respectively . In particular, with green excitation we could show that aceton fixation resulted in strong fluorescence of individual cells within a vast population of cells showing little or no autofluorescence . In addition, we could demonstrate that mounting media are able to strongly modify autofluorescence in fungi . Using digital image acquisition with a cooled CCD camera we were able to quantify the influence of different mounting media on fluorescence intensities of Aspergillus fumigatus.

Bone Marrow Transplant, 1998 Jul, 22 Suppl 1, S48 - 51
Detection of cell surface determinants for anti-Leu M3 (CD14), MY9 (CD33) and MY4 (CD14) and phagocytic function of cord blood monocytes in the course of gestational age; Gengenbacher D et al.; To investigate immunological functions of cord blood cells, cord blood monocytes of 84 term and preterm newborn infants were examined in presenting cell surface determinants for the monoclonal antibodies anti-Leu M3 (CD14), My9 (CD33) and My4 (CD14) using an alkaline-phosphatase-anti-alkaline-phosphatase staining method . We also tested the phagocytic function of these cells by incorporation of Candida albicans in a 60 min incubation assay . Mean values for antigen expression ranged from 60 to 70%, whereby My4 showed a statistically significant higher level compared to Anti-Leu M3 and My9 (P = 0.05) . Changes of antigen expression were statistically significant by the time of maturation . Statistically significant differences between male and female newborns were observed in 26-33 and 36-37 weeks of gestation . Considering the modus of delivery, no significantly higher amount of antigen-presenting monocytes were found in newborns of spontaneous birth compared to cesarean sections . However, no statistically significant differences in phagocytosis were observed between term and preterm newborn infants, mature newborns showed higher levels of both antigen expressing and phagocytic monocytes in contrast to a higher number of phagocytic cord blood monocytes only in premature infants.

Mycoses, 1998 May-Jun, 41(5-6), 219 - 21
Extensive skin candidosis in an adult: effective treatment with itraconazole; Daning L et al.; A 34-year-old man with 6 years' untreated erythematous scaling of the skin was diagnosed as having extensive skin candidosis . Oral itraconazole was administered for 4 weeks (400 mg day-1 for the first 3 weeks and 200 mg day-1 for the last week) . At the end of the 4-week treatment period, the rash was completely cleared from all skin sites, and Candida albicans was absent in culture tests on tissue samples . No adverse events were reported by the patient, and laboratory analysis revealed no abnormalities in the liver, kidney and haematological systems . Oral itraconazole is therefore an effective treatment for extensive skin candidosis.

Gene, 1998 Jul 30, 215(2), 311 - 8
Cloning and characterization of mitochondrial methionyl-tRNA synthetase from a pathogenic fungi Candida albicans; Lee SW et al.; A genomic sequence encoding mitochondrial methionyl-tRNA synthetase (MetRS) was determined from a pathogenic fungi Candida albicans . The gene is distinct from that encoding the cytoplasmic MetRS . The encoded protein consists of 577 amino acids (aa) and contains the class I defining sequences in the N-terminal domain and the conserved anticodon-binding amino acid, Trp, in the C-terminal domain . This protein showed the highest similarity with the mitochondrial MetRSs of Saccharomyces cerevisiae and Shizosaccharomyces pombe . The mitochondrial MetRSs of these fungi were distinguished from their cytoplasmic forms . The protein lacks the zinc binding motif in the N-terminal domain and the C-terminal dimerization appendix that are present in MetRSs of several other species . Escherichia coli tRNAMet was a substrate for the encoded protein as determined by genetic complementation and in vitro aminoacylation reaction . This cross-species aminoacylation activity suggests the conservation of interaction mode between tRNAMet and MetRS.

Infect Immun, 1998 Sep, 66(9), 4440 - 9
CD86 (B7-2), but not CD80 (B7-1), expression in the epidermis of transgenic mice enhances the immunogenicity of primary cutaneous Candida albicans infections; Gaspari AA et al.; Transgenic (Tg) mice whose epidermal keratinocytes constitutively overexpress either B7-1 (CD80) or B7-2 (CD86) exhibited exaggerated cutaneous delayed type hypersensitivity (DTH) to haptens compared to non-Tg mice . To determine whether enhanced DTH in these Tg mice is seen in response to cutaneous fungal infections, a primary infection with Candida albicans was established by inoculating this organism on the occluded skin of Tg and non-Tg mice . These infections resolved 7 days after removal of occlusive dressing in all three groups of mice, without evidence of exaggerated inflammation in either the Tg or non-Tg mice . Only B7-2 Tg mice developed enhanced Th1-lymphocyte-mediated immune responses to C . albicans antigens after resolving this infection: enhanced footpad swelling in response to intradermal C . albicans antigens, enhanced production of mRNA encoding Th1 lymphokines in draining lymph nodes, and increased gamma interferon secreted into culture supernatants by lymph node T lymphocytes stimulated with Candida antigens in vitro . Lastly, Western blotting of sera from mice that had resolved this fungal infection indicated that only B7-2 Tg mice recognized a wide range of Candida-associated antigens . These data suggest that these two costimulatory molecules, when expressed by keratinocytes, do not deliver identical signals to C . albicans antigen-reactive Th1 lymphocytes . The enhanced immune response in B7-2 Tg mice to a cutaneous C . albicans infection demonstrates the importance of antigen presentation and costimulation in immune reactivity to fungi . Furthermore, B7-2 Tg mice may be useful in identification of protective Candida antigens.

Infect Immun, 1998 Sep, 66(9), 4331 - 9
Coiling phagocytosis of trypanosomatids and fungal cells; Rittig MG et al.; Coiling phagocytosis has previously been studied only with the bacteria Legionella pneumophila and Borrelia burgdorferi, and the results were inconsistent . To learn more about this unconventional phagocytic mechanism, the uptake of various eukaryotic microorganisms by human monocytes, murine macrophages, and murine dendritic cells was investigated in vitro by video and electron microscopy . Unconventional phagocytosis of Leishmania spp . promastigotes, Trypanosoma cruzi trypomastigotes, Candida albicans hyphae, and zymosan particles from Saccharomyces cerevisiae differed in (i) morphology (rotating unilateral pseudopods with the trypanosomatids, overlapping bilateral pseudopods with the fungi), (ii) frequency (high with Leishmania; occasional with the fungi; rare with T . cruzi), (iii) duration (rapid with zymosan; moderate with the trypanosomatids; slow with C . albicans), (iv) localization along the promastigotes (flagellum of Leishmania major and L . aethiopica; flagellum or posterior pole of L . donovani), and (v) dependence on complement (strong with L . major and L . donovani; moderate with the fungi; none with L . aethiopica) . All of these various types of unconventional phagocytosis gave rise to similar pseudopod stacks which eventually transformed to a regular phagosome . Further video microscopic studies with L . major provided evidence for a cytosolic localization, synchronized replication, and exocytic release of the parasites, extending traditional concepts about leishmanial infection of host cells . It is concluded that coiling phagocytosis comprises phenotypically similar consequences of various disturbances in conventional phagocytosis rather than representing a single separate mechanism.

FEMS Microbiol Lett, 1998 Aug 1, 165(1), 103 - 9
Identification of four chitin synthase genes in the rice blast disease agent Magnaporthe grisea; Vidal-Cros A et al.; Four chitin synthase gene fragments were isolated from Magnaporthe grisea by use of polymerase chain reaction . A pair of degenerate primers based on conserved zymogen-type chitin synthase sequences amplified a approximately 600-bp product containing three chitin synthase gene fragments . A second pair of degenerate primers, based on conserved sequences between the chitin synthases 3 of Saccharomyces cerevisiae and Candida albicans, amplified a single 770-bp fragment . The four corresponding amino acid sequences each fell into one of the chitin synthase classes I-IV, as deduced from sequence analysis . Northern analysis demonstrated that the class II gene was expressed transiently in early phases of growth, whereas the class I and III genes as well as the class IV gene were expressed throughout the entire life cycle . Furthermore, the class III gene was the most expressed.

J Clin Pathol, 1998 May, 51(5), 390 - 1
A novel technique for assessment of adherence of Candida albicans to solid surfaces; Williams DW et al.; A novel approach for the assessment of adherence of Candida albicans to translucent acrylic material is described . The method uses the inverted microscope to visualise yeast adhering to acrylic surfaces while the test material remains immersed in buffer . Adherent cells were not subjected to surface tension forces that can occur during drying processes, so that an even distribution of yeast with no aggregation occurred . The process of counting attached yeast was subsequently performed without difficulty . From the 11 C albicans isolates examined, two groups were evident with respect to acrylic adherence: one group of four isolates with an adherence level of 400 yeast/mm2 acrylic, and one group of seven isolates with adherence levels of 1000 yeast/mm2 acrylic.

Aust Dent J, 1998 Jun, 43(3), 160 - 6
Candida-associated denture stomatitis . Aetiology and management: a review . Part 2 . Oral diseases caused by Candida species; Webb BC et al.; Certain systemic conditions and/or defects in the immune system may predispose the host to oral candidal infection and the commonest form of oral candidosis is candida-associated denture stomatitis . Until recently there has been controversy concerning the aetiology of the disease . Although some earlier investigators linked denture stomatitis with trauma or bacterial infection, others had isolated Candida albicans from the mouths of patients with the condition . Current studies indicate that denture stomatitis lesions are associated with the detection of candida species while other factors such as denture hygiene, trauma, systemic diseases and deficiencies of the immune system may be involved.

Proc Natl Acad Sci U S A, 1998 Aug 18, 95(17), 9825 - 30
The Candida albicans KRE9 gene is required for cell wall beta-1, 6-glucan synthesis and is essential for growth on glucose; Lussier M et al.; We have isolated CaKRE9, a gene from Candida albicans, that is a functional homologue of the Saccharomyces cerevisiae KRE9 gene involved in beta-1,6-glucan synthesis . Disruption of the CaKRE9 gene in C . albicans shows that CaKre9p is required for the synthesis or assembly of this fungal polymer . Homozygous null disruptants of CaKRE9 grow poorly on galactose and fail to form hyphae in serum, and, in growth medium containing glucose, the gene is essential . Thus, the CaKRE9 gene product is a potentially useful candidate as a target for fungal-specific drugs.

Am J Health Syst Pharm, 1998 Aug 1, 55(15), 1584 - 7
Microbial inhibitory properties and stability of topotecan hydrochloride injection; Patel K et al.; The viability of five microorganisms in topotecan 1 mg/mL (as the hydrochloride salt) in sterile water and the stability of the drug were studied . Duplicate portions of topotecan 1 mg/mL were inoculated with Escherichia coli . The process was repeated for Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Aspergillus niger . Samples were removed from each solution initially and after 6, 16, and 24 hours and 3, 7, 14, 21, and 28 days of incubation at 20-25 degrees C . To test stability, vials of reconstituted topotecan hydrochloride injection were stored at each of three temperatures--5, 25, and 30 degrees C--and other vials were used for time zero analysis . For each temperature, vials were removed at 1, 7, and 14 days and the remaining vials at 28 days for analysis by high-performance liquid chromatography and for visual and pH assessment . P . aeruginosa, S . aureus, and E . coli lost viability at 16 hours, 24 hours, and 28 days, respectively . C . albicans and A . niger did not lose viability, but their numbers did not grow . No differences in color or clarity were observed, and pH was constant . In all solutions, the topotecan concentration was > 98% of the initial concentration . Topotecan 1 mg/mL in sterile water stored at 20-25 degrees C for up t 28 days did not support growth of the five microorganisms studied; in solutions stored at 5, 25, or 30 degrees C for up to 28 days, topotecan 1 mg/mL remained stable.

Eur Surg Res, 1998, 30(4), 290 - 6
Measurement of (1-->3)-beta-D-glucan in an experimental model of systemic candidiasis; Kawagoe T et al.; To investigate the utility of measuring blood concentrations of (1-->3)-beta-D-glucan, a component of the fungal cell wall, as an auxiliary diagnostic method for systemic candidiasis, rats were inoculated with Candida albicans and the number of C . albicans in the viscera and glucan in the blood were quantitated . The concentration of blood glucan and the number of C . albicans in the viscera were also measured both under leukopenia and with deteriorated reticuloendothelial system cell function, and when the liver and spleen had been excised . As a result, systemic candidiasis appeared in the group with leukopenia, and the number of living C . albicans increased in the kidney and liver . Together with this increase in the number of C . albicans, there was an increase in blood (1--> 3)-beta-D-glucan . Measurements of blood (1--> 3)-beta-D-glucan well reflect a proliferation of C . albicans in vivo, which would make this a useful auxiliary for the clinical diagnosis of systemic mycosis.

J Clin Microbiol, 1998 Sep, 36(9), 2690 - 5
Stable phenotypic resistance of Candida species to amphotericin B conferred by preexposure to subinhibitory levels of azoles; Vazquez JA et al.; The fungicidal activity of amphotericin B (AmB) was quantitated for several Candida species . Candida albicans and C . tropicalis were consistently susceptible to AmB, with less than 1% survivors after 6 h of exposure to AmB . C . parapsilosis and variants of C . lusitaniae and C . guilliermondii were the most resistant, demonstrating 50 to 90% survivors in this time period and as high as 1% survival after a 24-h exposure time . All Candida species were killed (<1% survivors) after 24 h of exposure to AmB . In contrast, overnight exposure to either fluconazole or itraconazole resulted in pronounced increases in resistance to subsequent exposures to AmB . Most dramatically, C . albicans was able to grow in AmB cultures after azole preexposure . Several other Candida species did not grow in AmB but showed little or no reduction in viability after up to 24 h in AmB . Depending on the growth conditions, Candida cells preexposed to azoles may retain AmB resistance for days after the azoles have been removed . If this in vitro antagonism applies to the clinical setting, treatment of patients with certain antifungal combinations may not be beneficial . The ability of some Candida isolates to survive transient exposures to AmB was not reflected in the in vitro susceptibility changes as measured by standard MIC assays . This finding should be considered in studies attempting to correlate patient outcome with in vitro susceptibilities of clinical fungal isolates . Patients who fail to respond to AmB may be infected with isolates that are classified as susceptible by standard in vitro assays but that may be resistant to transient antifungal exposures which may be more relevant in the clinical setting.

J Ethnopharmacol, 1998 Jul, 61(3), 237 - 41
Antimicrobial and anticonvulsant activities of Viscum capense; Amabeoku GJ et al.; Dichloromethane, methanol and water extracts of Viscum sapense L.f., of the Loranthaceae family, were tested for antimicrobial activities against Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans . Methanol extract was also tested for activity against seizures in albino mice induced by pentylenetetrazole (PTZ), bicuculline and N-methyl-DL-aspartic acid (NMDLA) . Methanol extract of V . capense inhibited the growth of S . aureus . Methanol extract also protected the mice against PTZ- and bicuculline-induced tonic seizures but did not significantly alter NMDLA-induced tonic seizures . The data indicate that the extract of V . capense has antibacterial activity against S . aureus and also anticonvulsant activity.

Gynecol Obstet Invest, 1998 Aug, 46(2), 73 - 4
Breast-feeding, pain and infection; Thomassen P et al.; A syndrome of deep pain in the breast during and immediately after lactation has been ascribed to an infection with Candida albicans . A series of 20 patients with deep pain, another 20 with superficial infection and 20 healthy women were compared with respect to the growth of bacteria and fungi . C . albicans was found twice as often in the milk of women with superficial lesions compared to those with deep pain . Bacteria were often found on the nipple and in the milk of those complaining of deep pain . Thus, if the deep pain syndrome is caused by microbes, this study points to a pathogenic role of bacteria rather than fungi.

Zh Mikrobiol Epidemiol Immunobiol, 1998 May-Jun, (3), 51 - 5
{Proliferative response and suppressor activity of lymphocytes in candidiasis}; Sardyko NV et al.; In 14 practically healthy donors, 22 patients with chronic candidiasis of the skin and mucosa (CCSM) and 14 patients with chronic candidal vulvovaginitis (CCW) immunoregulating factors synthesized by peripheral blood lymphocytes under the conditions of in vitro induction of suppressor cells were studied in comparison with the proliferative response to PHA and Candida albicans cytoplasmic protein . In practically healthy donors the level of lymphocyte response to mitogen coincided with the presence of the inhibiting activity of supernatants . In a more severe form of Candida infection (CCSM) the proliferative reaction of lymphocytes to both PHA and C.albicans antigen was weakened or even inhibited . The level of the proliferative response of lymphocytes was inversely proportional to the immunoregulating activity of cells, nonstimulated and stimulated with ConA and C.albicans cytoplasmic protein . In CCW lymphocytes exhibited good proliferative response to T-mitogen, occurring in combination with weak reaction, or no reaction at all, to C.albicans antigen . A decrease in the level of spontaneous blast transformation was accompanied by an increase in spontaneous suppressor activity.

Dev Comp Immunol, 1998 Jul-Aug, 22(4), 387 - 99
A gloverin-like antibacterial protein is synthesized in Helicoverpa armigera following bacterial challenge; Mackintosh JA et al.; A bacteria inducible antibacterial protein, P2, was isolated from the old world bollworm Helicoverpa armigera . Fifth-instar larvae were injected with live Escherichia coli NCTC 8196 . P2 was isolated by HPLC using reversed-phase and size-exclusion columns . In addition, P2 was isolated by an alternative method of sequential cation-exchange and reversed-phase HPLC . The structure of P2 was determined by N-terminal Edman degradation and mass spectrometry . P2 had similar mass (14.1 kDa) structure and activity to gloverin, an inducible glycine-rich antibacterial protein isolated from Hyalophora gloveri {Axen, A.; Carlsson, A.; Engstrom, A.; Bennich, H . Eur . J . Biochem . 247:614-619; 1997} . At the N-terminus P2 had approximately 60% identity with gloverin . P2 is basic, heat stable, and displayed rapid antibacterial action . P2 was active against the Gram-negative bacteria tested and was inactive against the Gram-positive bacteria, Candida albicans, a bovine turbinate cell line, and pestivirus.

J Infect Dis, 1998 Aug, 178(2), 488 - 96
Recognition of antigenic clusters of Candida albicans by T lymphocytes from human immunodeficiency virus-infected persons; Kunkl A et al.; The fine specificity of the cellular immune response to Candida albicans (i.e., recognition of different antigenic components) between normal controls and human immunodeficiency virus-infected patients in various stages of disease was compared . C . albicans-specific T cells, enriched by antigen stimulation and interleukin-2 expansion, were challenged with antigenic fractions of different molecular weight obtained by SDS-gel fractionation of C . albicans extracts in the presence of autologous mononuclear cells as antigen-presenting cells . Proliferative responses showed similar patterns of reactivity between controls and category A and B seropositive subjects . Category C patients with concurrent C . albicans infections did not give rise to C . albicans-specific T cell lines, confirming the T cell defect . Patients without clinically evident C . albicans infection had a low but broad reactivity pattern of C . albicans-specific T cells . These results suggest that depletion of C . albicans-specific T cells, independent of their fine specificity, occurs along with disease progression.

J Infect Dis, 1998 Aug, 178(2), 478 - 87
Candidiasis in interferon-gamma knockout (IFN-gamma-/-) mice; Balish E et al.; Germ-free C57BL/6 x 129 interferon-gamma knockout (IFN-gamma(-/-)) mice and their immunocompetent (+/-, +/+) counterparts were colonized with a pure culture of Candida albicans to assess their natural susceptibility to mucosal and systemic candidiasis of endogenous origin . Colonization with a pure culture of C . albicans was not lethal for adult or neonatal IFN-gamma(-/-) gnotobiotic mice over the 15-week study . The IFN-gamma(-/-) mice were more susceptible to gastric (cardia-antrum section), anorectal, and acute systemic (intravenous challenge) candidiasis than immunocompetent controls, and some IFN-gamma(-/-) mice developed intestinal adenomas after colonization with C . albicans . The enhanced susceptibility of IFN-gamma(-/-) mice, compared with immunocompetent controls, may be associated with a poor proliferative response of spleen cells to C . albicans antigens and a T helper 2 (IgG1) serum antibody response to C . albicans antigens . Thus, IFN-gamma is important for murine resistance to gastric, anorectal, and acute systemic candidiasis.

Przegl Lek, 1998, 55(3), 133 - 5
{Use of Tienam in treatment of severe acute pancreatitis}; Pruszynski K et al.; Therapeutic results obtained in the management of severe acute pancreatitis (AP) complications are still unsatisfactory, and mortality rate is about 40% . An improvement in treatment effectiveness can be related to successful control of infections and prevention of uncontrollable sepsis . The aim of the study was evaluation of effectiveness of Tienam in the management of severe acute pancreatitis complications . The clinical material included 24 patients, treated for AP between 1994 and 1996 . All of the patients were operated on and 19 of them required at least one reoperation . During primary surgery we performed necrosectomy and continuous peritoneal lavage . The analysis of cultures obtained from infected necrotic tissue, pancreatic abscesses and whole blood showed that the infections were caused mainly by Gram-negative bacteria . The highest efficiency in infections control was observed in the case of Tienam (15% resistant strains) . The authors emphasize also the increasing role of Staphylococcus aureus and Candida albicans in secondary infections.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(3), 173 - 8
Evaluation of antifungal activity of an antifungal drug by in vitro simulation of in vivo pharmacokinetics of the drug against fungal hyphal growth; Oh KB et al.; An automatic drug concentration simulator (DCS) has been developed and its applicability has been demonstrated by in vitro simulation of the human plasma concentration-time curve of fluconazole (FLCZ) against hyphal growth of Candida albicans and Aspergillus fumigatus . The response of hyphal growth to FLCZ was continually monitored and analyzed using an automatic hyphal growth analyzing system (Bio-Cell Tracer) . The simulated concentration of FLCZ by DCS was confirmed by HPLC . The DCS assay was reproducible with a mean coefficient of variation (C.V., n=3) of 5.38 % . When the growth of C . albicans hyphae was tested, there was a lag of onset of FLCZ effect between the time when FLCZ concentration became maximal (C MAX, 7.95 microg/ml ) and the point at which hyphal growth ceased . In contrast, FLCZ was found inactive against A . fumigatus . The newly devised technique could provide clinicians with important information in determining optimal dosing regimens for antifungal drugs.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(3), 161 - 5
{Cooperative inhibitory effects of cervical mucus and neutrophils on growth of Candida albicans}; Okutomi T et al.; Preparations of human cervical mucus were obtained from fifteen female volunteers with or without vaginal candidiasis . Their content of lactoferrin and inhibitory effects on the growth of Candida albicans were estimated . The concentration of lactoferrin in cervical mucus was measured by enzyme linked immunosorbent assay (ELISA) . Lactoferrin concentration of the eleven preparations among them was more than 0.2 mg/ml . The effects of these cervical mucus preparations on Candida growth were examined in vitro . Candida growth was not inhibited by the addition of 1/200 diluted cervical mucus alone, however, anti-Candida activity of murine neutrophils was augmented by such an addition . These results suggest that the combination of neutrophils and cervical mucus may have an important role in defense against Candida infection in the vaginal mucosa.

J Biol Chem, 1998 Aug 14, 273(33), 20837 - 46
Multiple functions of Pmt1p-mediated protein O-mannosylation in the fungal pathogen Candida albicans; Timpel C et al.; Protein mannosylation by Pmt proteins initiates O-glycosylation in fungi . We have identified the PMT1 gene and analyzed the function of Pmt1p in the fungal human pathogen Candida albicans . Mutants defective in PMT1 alleles lacked Pmt in vitro enzymatic activity, showed reduced growth rates, and tended to form cellular aggregates . In addition, multiple specific deficiencies not known in Saccharomyces cerevisiae (including defective hyphal morphogenesis; supersensitivity to the antifungal agents hygromycin B, G418, clotrimazole, and calcofluor white; and reduced adherence to Caco-2 epithelial cells) were observed in pmt1 mutants . PMT1 deficiency also led to faster electrophoretic mobility of the Als1p cell wall protein and to elevated extracellular activities of chitinase . Homozygous pmt1 mutants were avirulent in a mouse model of systemic infection, while heterozygous PMT1/pmt1 strains showed reduced virulence . The results indicate that protein O-mannosylation by Pmt proteins occurs in different fungal species, where PMT1 deficiency can lead to defects in multiple cellular functions.

Genetics, 1998 Aug, 149(4), 1739 - 52
A physical map of chromosome 7 of Candida albicans; Chibana H et al.; As part of the ongoing Candida albicans Genome Project, we have constructed a complete sequence-tagged site contig map of chromosome 7, using a library of 3840 clones made in fosmids to promote the stability of repeated DNA . The map was constructed by hybridizing markers to the library, to a blot of the electrophoretic karyotype, and to a blot of the pulsed-field separation of the SfiI restriction fragments of the genome . The map includes 149 fosmids and was constructed using 79 markers, of which 34 were shown to be genes via determination of function or comparison of the DNA sequence to the public databases . Twenty-five of these genes were identified for the first time . The absolute position of several markers was determined using random breakage mapping . Each of the homologues of chromosome 7 is approximately 1 Mb long; the two differ by about 20 kb . Each contains two major repeat sequences, oriented so that they form an inverted repeat separated by 370 kb of unique DNA . The repeated sequence CARE2/Rel2 is a subtelomeric repeat on chromosome 7 and possibly on the other chromosomes as well . Genes located on chromosome 7 in Candida are found on 12 different chromosomes in Saccharomyces cerevisiae.

J Biol Chem, 1998 Aug 7, 273(32), 20438 - 47
Candidacidal activity of salivary histatins . Identification of a histatin 5-binding protein on Candida albicans; Edgerton M et al.; Candida albicans is the predominant species of yeast isolated from patients with oral candidiasis, which is frequently a symptom of human immunodeficiency virus infection and is a criterion for staging and progression of AIDS . Salivary histatins (Hsts) are potent in vitro antifungal agents and have great promise as therapeutic agents in humans with oral candidiasis . The molecular mechanisms by which Hsts kill yeast cells are not known . We report here, that unlike other antimicrobial proteins, Hsts do not display lytic activities to lipid membranes, measured by release and dequenching of the fluorescent dye calcein . Analysis of the magnitude and time course of Hst-induced calcein release from C . albicans cells further showed that loss of cell integrity was a secondary effect following cell death, rather than the result of primary disruption of the yeast cell membrane . 125I-Hst 5 binding studies indicated that C . albicans expressed a class of saturable binding sites (KD = 1 microM), numbering 8.6 x 10(5) sites/cell . Both Hst 3 and Hst 4 competed for these binding sites with similar affinities, which is consistent with the micromolar concentration of Hsts required for candidacidal activity . Specific 125I-Hst 5 binding was not detected to C . albicans spheroplasts, which were 14-fold less susceptible to Hst 5 killing, compared with intact cells in candidacidal assays . In overlay experiments, 125I-Hst 5 bound to a 67-kDa protein detected in C . albicans whole cell lysates and crude membrane fractions, but not in the yeast cell wall fraction . Consistent with the overlay data, cross-linking of 125I-Hst 5 to C . albicans resulted in the appearance of a specific 73-kDa 125I-Hst 5-containing complex that was not detected in the cell wall . 125I-Hst 5-binding protein of similar size was also observed in susceptible S . cerevisiae strain TI#20 . This is the first description of Hst 5 binding sites on C . albicans which mediate cell killing and identification of a 67-kDa yeast Hst 5-binding protein . The binding characteristics of Hst 5 are in agreement with the observed potency of its biological effect and provide crucial information to the use of Hst 5 as a therapeutic agent . The presence of a specific C . albicans Hst 5-binding protein provides further insight into the potential mechanism of yeast killing and suggests a basis for differential activity between yeast killing and the nontoxic nature of Hsts to humans.

Appl Microbiol Biotechnol, 1998 Jun, 49(6), 766 - 9
Inactivation of bacteriophage lambda, Escherichia coli, and Candida albicans by ozone
Komanapalli IR, Lau BH.
The effects of ozone (O3) on three types of microbes were studied . Test suspensions were exposed to 600 ppm O3 at room temperature . Control experiments were performed under identical conditions using oxygen gas . Bacteriophage lambda was completely inactivated at 10 min while Escherichia coli and Candida albicans were only inactivated by factors of 10(5) and 10(4) respectively at 40 min . Exposure of a mixed microbial suspension to O3 for 5 min resulted in 100% killing of bacteriophages while the viability of E . coli remained unchanged . Various body fluids containing phages were exposed to O3 . Compared to buffered solution, the decrease in phage titers was significantly slower in whole blood, plasma, and albumin . Both E . coli and C . albicans had increased production of thiobarbituric-acid-reactive substances with increased O3 exposure . 3H-labelled amino acids were incorporated into E . coli . O3 treatment resulted in a loss of radioactivity, indicating leakage of cytoplasmic contents . The data indicate that microbes are inactivated by O3 at different rates, possibly related to differential membrane permeability . The milieu in which microbes are present determines the effectiveness and outcome of O3 treatment.

Arch Microbiol, 1998 Aug, 170(2), 113 - 9
Candida albicans HEX1 gene, a reporter of gene expression in Saccharomyces cerevisiae; Niimi K et al.; The Candida albicans HEX1 gene was examined as a reporter of gene expression in Saccharomyces cerevisiae . The galactose-inducible S . cerevisiae GAL1-GAL10 promoter was inserted upstream of the C . albicans HEX1 gene, which encodes N-acetylglucosaminidase . The gene was introduced into S . cerevisiae AH22, which has no background N-acetylglucosaminidase activity . Expression of HEX1 in transformed cells was induced significantly by galactose and was repressed by glucose . The HEX1 gene product was functional in S . cerevisiae cells and was targeted to the periplasm . Both untransformed S . cerevisiae cells and cells expressing HEX1 had similar growth curves and cell morphology indicating that expression of N-acetylglucosaminidase was not toxic to the host strain . These results demonstrate that the HEX1 gene can be a useful reporter of gene expression in S . cerevisiae.

J Bacteriol, 1998 Aug, 180(15), 3809 - 15
Disruption of the Candida albicans TPS1 gene encoding trehalose-6-phosphate synthase impairs formation of hyphae and decreases infectivity; Zaragoza O et al.; The TPS1 gene from Candida albicans, which encodes trehalose-6-phosphate synthase, has been cloned by functional complementation of a tps1 mutant from Saccharomyces cerevisiae . In contrast with the wild-type strain, the double tps1/tps1 disruptant did not accumulate trehalose at stationary phase or after heat shock . Growth of the tps1/tps1 disruptant at 30 degreesC was indistinguishable from that of the wild type . However, at 42 degreesC it did not grow on glucose or fructose but grew normally on galactose or glycerol . At 37 degreesC, the yeast-hypha transition in the mutant in glucose-calf serum medium did not occur . During growth at 42 degreesC, the mutant did not form hyphae in galactose or in glycerol . Some of the growth defects observed may be traced to an unbalanced sugar metabolism that reduces the cellular content of ATP . Mice inoculated with 10(6) CFU of the tps1/tps1 mutant did not show visible symptoms of infection 16 days after inoculation, while those similarly inoculated with wild-type cells were dead 12 days after inoculation.

J Bacteriol, 1998 Aug, 180(15), 3771 - 8
Lack of consistent short sequence repeat polymorphisms in genetically homologous colonizing and invasive Candida albicans strains; Lunel FV et al.; Short sequence repeats (SSRs), potentially representing variable numbers of tandem repeat (VNTR) loci, were identified for the human-pathogenic yeast species Candida albicans by computerized DNA sequence scanning . The individual SSR regions were investigated in different clinical isolates of C . albicans . Most of the C . albicans SSRs were identified as genuine VNTRs . They appeared to be present in multiple allelic variants and were demonstrated to be diverse in length among nonrelated strains . As such, these loci provide adequate targets for the molecular typing of C . albicans strains . VNTRs encountered in other microbial species sometimes participate in regulation of gene expression and function as molecular switches at the transcriptional or translational level . Interestingly, the VNTRs identified here often encode polyglutamine stretches and are frequently located within genes potentially involved in the regulation of transcription . DNA sequencing of these VNTRs demonstrated that the length variability was restricted to the CAA/CAG repeats encoding the polyglutamine stretches . For these reasons, paired C . albicans isolates of similar genotype, either found as noninvasive colonizers or encountered in an invasive state in the same individual, were studied with respect to potentially invasion-related alterations in the VNTR profiles . However, none of the VNTRs analyzed thus far varied systematically with the transition from colonization to invasion . In contrast to the situation described for some prokaryotic species, this finding suggests that VNTRs of C . albicans may not simply function as contingency loci related to straightforward on/off regulation of invasion-related gene expression.

J Oral Pathol Med, 1998 May, 27(5), 213 - 9
The effect of limited exposure to antifungal agents on the germ tube formation of oral Candida albicans; Ellepola AN et al.; Candidal adherence has been implicated as the first step in the pathogenesis of oral candidosis, and germ tube formation by Candida albicans has been attributed as a co-factor that promotes adherence . Oral candidosis is treated with polyenes and the azole group of antifungal agents . As the intraoral concentrations of antifungals fluctuate considerably due to the dynamics of the oral cavity, we investigated the effect of short exposure to sub-lethal concentrations of antifungals on the germ tube formation of Candida albicans . After determining the minimum inhibitory concentration (MIC) of the antifungal agents, ten oral isolates of Candida albicans were exposed to sub-lethal concentrations of nystatin (6xMIC), amphotericin B (8xMIC), 5-fluorocytosine (8xMIC), ketoconazole (4xMIC) and fluconazole (4xMIC), for 1 h . Following removal of the antifungal agent and subsequent incubation in a germ tube-inducing medium, the germ tube formation of these isolates was quantified . When compared with the controls, exposure to nystatin and amphotericin B almost completely inhibited germ tube formation of all the isolates (mean percentage reduction of 97.68 and 97.52%, respectively; P<0.0001), while ketoconazole suppressed this activity to a lesser degree (30.84%; P=0.0174) . However, 5-fluorocytosine- and fluconazole-mediated germ tube suppression was minimal (12.63 and 15.93%, respectively; P=0.3255 and P=0.3791) . In clinical terms, these findings indicate that short exposure to sub-therapeutic levels of commonly prescribed antifungals may modulate candidal germ tube formation, and thereby the clearance of the organisms from the oral cavity.

Immunol Cell Biol, 1998 Jun, 76(3), 209 - 16
The regulation of burn-associated infections with herpes simplex virus type 1 or Candida albicans by a non-toxic aconitine-hydrolysate, benzoylmesaconine . Part 2: Mechanism of the antiviral action; Kobayashi M et al.; In the accompanying paper, the resistance to infections with HSV type 1 (HSV-1) and Candida albicans was improved in thermally injured mice treated with benzoylmesaconine (BEN), an aconitine-hydrolysate isolated from heated Aconiti tuber, or inoculated with splenic CD4+ T cells from BEN-treated mice (BEN T cells) . In this paper, therefore, the antiviral mechanism of BEN T cells (or BEN) on the improved resistance of burned mice to the HSV-1 infection was studied . Burn-associated CD + CD11b+ TCRgamma/delta+ type-2 T cells have been shown to be a key on the increased susceptibility of thermally injured mice to infection with HSV-1 or C . albicans . The susceptibility of T6S-mice, mice inoculated with 1 x 10(6) cells/mouse of T6S cells (a clone of burn-associated type-2 T cells), to HSV-1 infection was similar to that of thermally injured mice . The adoptive transfer of BEN T cells to T6S-mice restores their impaired resistance to HSV-1 infection . The type-2 cytokine levels in sera of T6S-mice were decreased after inoculation of BEN T cells . BEN T cells inhibited the type-2 cytokine production by T6S cells when they were cocultured in vitro . BEN T cells, characterized as CD4+ CD28+ TCRalpha/beta+ Vicia villosa (VV) lectin-adherent T cells, showed non-specific ability to inhibit the cytokine production by various type-2 T cells . From the results of the cytokine-producing profile, BEN T cells were shown to be a different subset of CD4+ T cells from Th1 and Th2 cells, although these three CD4+ T cells had similar properties phenotypically . BEN T cells were induced in normal mice 1-4 days after the oral treatment of BEN (1 microg/kg or more) . These results suggest that, through the induction of antagonistic CD4+ T cells against burn-associated type-2 T cells, BEN may improve the resistance of T6S-mice (or thermally injured mice) to the infection of HSV-1.

Immunol Cell Biol, 1998 Jun, 76(3), 202 - 8
The regulation of burn-associated infections with herpes simplex virus type 1 or Candida albicans by a non-toxic aconitine-hydrolysate, benzoylmesaconine . Part 1: Antiviral and anti-fungal activities in thermally injured mice; Kobayashi M et al.; As compared with normal unburned mice, thermally injured mice have been shown to be 50-100 times more susceptible to HSV type 1 (HSV-1) or Candida albicans infection . Benzoylmesaconine (BEN) improved the resistance of thermally injured mice against infection with HSV-1 or C . albicans to the level observed in normal mice . Mortality rates of normal mice exposed to lethal amounts of these pathogens were not affected by the BEN treatment, while significant survival effects were produced in these mice after treatment with acyclovir (against HSV-1) or amphotericin B (against C . albicans) . Benzoylmesaconine did not inhibit the growth of these pathogens in vitro and did not directly reduce the viability of the pathogens . However, burned mice inoculated with CD4+ T cells from BEN-treated mice resisted infections from these pathogens . These results suggested that, through the generation of CD4+ T cells, BEN recovered the impaired resistance of thermally injured mice to infection by HSV-1 or C . albicans.

Spine, 1998 Jul 15, 23(14), 1600 - 6
Percutaneous suction aspiration and drainage for pyogenic spondylitis; Nagata K et al.; STUDY DESIGN: Retrospective evaluation of results in 23 cases of early-stage pyogenic spondylitis treated with percutaneous suction aspiration and drainage . OBJECTIVES: To evaluate the efficacy of percutaneous suction aspiration and drainage as a treatment method for early-stage pyogenic spondylitis . SUMMARY OF BACKGROUND DATA: Traditional surgical treatment for pyogenic spondylitis has the disadvantage of increased morbidity caused by the extensive exposure required in the presence of infection . Recently, a few case reports have described minimally invasive treatment for pyogenic spondylitis in which percutaneous suction aspiration was used . However, the efficacy of this new treatment has not yet been evaluated . METHODS: All charts, radiographs, and bacteriologic and histologic findings were reviewed . All 23 patients who received the new treatment were observed clinically and radiographically, to evaluate the efficacy of the treatment . RESULTS: To date, all patients have been observed for more than 2 years . Twenty (87%) of the 23 patients have shown good results according to the evaluation . The causative organism was identified using tissue culturing in 12 (52%) of the 23 patients . The causative organism was Staphylococcus aureus in 8, and Staphylococcus epidermidis, Candida albicans, Pseudomonas aeruginosa, and Propionibacterium acnes in 1 each . Back pain as the major symptom in these patients was relieved within an average of 9.4 days after the operation . However, the patient in whom the spondylitis was caused by Candida albicans has received this new treatment twice without success . CONCLUSIONS: Evaluation of percutaneous suction aspiration with drainage shows that it is an effective treatment for early-stage pyogenic spondylitis.

J Biol Regul Homeost Agents, 1998 Jan-Jun, 12(1-2), 33 - 7
Decreased DTH response to recall antigens in mice injected with human immunodeficiency virus type 1-infected U937 cells and infected with Candida albicans; Lande R et al.; In previous studies, we have reported that the intraperitoneal (i.p.) injection of HIV-1 infected human U937 cells into normal mice resulted in long-term persistence of anti-HIV antibodies and in a small percentage (10-20%) of HIV-1 infected animals at 6-12 months after the injection . The study reported here was undertaken to detect T immune defects in U937-HIV-1-injected mice . Eight months after the initial injection, a marked decrease in DTH response against U937 cells was detected in HIV injected animals . In addition, a consistent decrease in DTH response against a soluble mannoprotein antigen of Candida albicans cell wall (MP-F2) was also observed in U937-HIV-1-injected mice, chronically infected with low-virulent strain of the fungus . No decreases in DTH response was observed in control-injected animals . These data indicate that U937-HIV-1-injected mice become unable to mount a normal antigen-specific immune response . Although the mechanisms involved in the generation of these T cell defects remain unclear, these events appear to be somehow related to the HIV-1 infection and should be considered in the current studies of HIV-1 infection with transgenic mice.

FEMS Microbiol Lett, 1998 Jun 15, 163(2), 121 - 7
Adhesion of Candida albicans mutant strains to host tissue; Arie ZR et al.; Adhesion of Candida albicans to host cells is believed to represent a fungal virulence factor and a significant step in the development of candidiasis . As C . albicans strains may differ in their in vitro adhesion ability we initiated a study to investigate whether mutant strains differ in this respect from their parent wild-type . We assessed the in vitro adhesion of C . albicans CBS562 and two mutants obtained by mutagenesis with N'-nitrosoguanidine: a histidine auxotroph, SAG5, derived from CBS562, and a respiratory-deficient strain (a petite mutant), SAR1, derived from SAG5 . The adhesion was tested in vitro using two target cell systems: (1) exfoliated human buccal epithelial cells (BEC); and (2) human keratinocyte tissue line cells (HaCaT cells) . Adhesion to BEC was evaluated microscopically and that to HaCaT cells by a direct ELISA technique . The results indicated a 54% reduction in adhesion to BEC for SAG5 and 30% for SAR1 as compared to the wild-type, and a 25% reduction in adhesion to HaCaT cells for SAG5 and 20% for SAR1 . To verify whether the prototrophy restores the adhesion ability, we complemented the his-negative auxotroph by transforming the strain with the HIS4 gene . Then we assayed the adhesion to BEC of the complemented his-negative mutant in comparison to that of the wild-type, the his-negative mutant (SAG5) and the plasmid-cured transformant . The adhesion values of the complemented his-negative strain were similar to those of the wild-type, whereas the values of the plasmid-cured strain were similar to those of SAG5.

J Antibiot (Tokyo), 1998 May, 51(5), 487 - 91
Antifungal activities of rapamycin and its derivatives, prolylrapamycin, 32-desmethylrapamycin, and 32-desmethoxyrapamycin; Wong GK et al.; The antifungal agent rapamycin is highly effective in inhibiting growth of yeast and mold strains . This study demonstrates that in liquid medium, rapamycin is more active than its derivatives (prolylrapamycin, 32-desmethylrapamycin, 32-desmethoxyrapamycin) against Candida albicans, Saccharomyces cerevisiae, and Fusarium oxysporum . All the rapamycins were more active than amphotericin B . Although four other molds were not inhibited in liquid medium, they were very sensitive to rapamycin and its derivatives when tested on agar . The latter assay showed that rapamycin is the most active and 32-desmethylrapamycin is more active than prolylrapamycin and 32-desmethoxyrapamycin . The conclusion of this study is that rapamycin is the most active antifungal agent of the compounds examined . The unexpected finding of high activity of rapamycin and its derivatives against filamentous fungi when assayed by the agar diffusion assay suggests that rapamycin or a derivative may hold promise for chemotherapy against pathogenic molds as well as yeasts.

Clin Microbiol Rev, 1998 Jul, 11(3), 415 - 29
Onychomycosis: pathogenesis, diagnosis, and management; Elewski BE; Although not life-threatening, onychomycosis (a fungal infection of the nail, usually caused by a dermatophyte) constitutes an important public health problem because of its high prevalence (about 10% of the U.S . population) and associated morbidity . The disease can have certain negative consequences for patients, such as pain, and can potentially undermine work and social lives . This review discusses the etiology, classification, diagnosis, and treatment of onychomycosis . Four types of onychomycosis are recognized based on the site and pattern of fungal invasion . Dermatophyte fungi are the predominant pathogens, but yeasts (especially Candida albicans) and nondermatophyte molds may also be implicated . Accurate diagnosis requires direct microscopy and fungal culture . The differential diagnosis includes psoriasis, lichen planus, onychogryphosis, and nail trauma . Onychomycosis is more difficult to treat than most dermatophytoses because of the inherent slow growth of the nail . Older antifungal agents (ketoconazole and griseofulvin) are unsuitable for onychomycosis because of their relatively poor efficacy and potential adverse effects . Three recently developed antimycotic agents (fluconazole, itraconazole, and terbinafine) offer high cure rates and good safety profiles . In addition, the short treatment times (< 3 months) and intermittent dosing schedules are likely to enhance compliance and reduce the costs of therapy.

Clin Diagn Lab Immunol, 1998 Jul, 5(4), 574 - 7
Comparison of inflammatory events during developing immunoglobulin E-mediated late-phase reactions and delayed-hypersensitivity reactions; Zweiman B et al.; To compare cellular and mediator responses in early developing late-phase skin reactions (LPR) and delayed-hypersensitivity (DH) reactions in the same subjects, responses in skin chambers overlying sites of challenge with pollen antigen and Candida albicans antigens were compared in six humans with demonstrated prominent LPR and DH responses . Histamine levels in overlying chamber fluids at 1 h were much higher at LPR than at DH sites (P = 0.002) . After the next 4 h, leukocyte exudation was higher at LPR than at DH sites (P = 0.005) . Most leukocytes were activated neutrophils with greater frequency of superoxide-secreting cells and released lactoferrin at LPR than at DH sites (P = 0.01 and P = 0.02, respectively) . The frequency of exuding eosinophils was higher, but not significantly so (P = 0.5), at LPR sites . Although significantly more eosinophils at LPR sites were activated (P = 0.02), the levels of released eosinophilic cationic protein were not significantly higher at LPR sites (P = 0.09) . The levels of interleukin-8 (IL-8), but not IL-6, were greater at LPR than at DH sites . During the first 5 h of challenge there was greater mast cell activation and subsequent exudation of activated neutrophils at sites of developing LPR than at DH sites, possibly related to greater local IL-8 levels . The frequency of activated eosinophils was also greater at LPR sites . These different initial inflammatory responses could play a role in determining expression of LPR or DH reactions.

Antimicrob Agents Chemother, 1998 Jul, 42(7), 1587 - 91
Inhibition of hyphal growth of azole-resistant strains of Candida albicans by triazole antifungal agents in the presence of lactoferrin-related compounds; Wakabayashi H et al.; The effects of bovine lactoferrin (LF) or the LF-derived antimicrobial peptide lactoferricin B (LFcin B) on the growth of Candida albicans hyphae, including those of three azole-resistant strains, were investigated by a crystal violet staining method . The hyphae of two highly azole-resistant strains were more susceptible to inhibition by LF or LFcin B than the azole-susceptible strains tested . One moderately azole-resistant strain was defective in the formation of hyphae and showed a susceptibility to LF greater than that of the susceptible strains but a susceptibility to LFcin B similar to that of the susceptible strains . The highly azole-resistant strain TIMM3317 showed trailing growth in the presence of fluconazole or itraconazole, while the extent of growth was reduced by the addition of LF or LFcin B at a sub-MIC . Thus, the addition of LF or LFcin B at a sub-MIC resulted in a substantial decrease in the MICs of fluconazole and itraconazole for two highly azole-resistant strains; e.g., the MIC of fluconazole for TIMM3317 was shifted from > 256 to 0.25 micrograms/ml by LF, but the MICs were not decreased for the susceptible strains . The combination effects observed with triazoles and LF-related compounds in the case of the two highly azole-resistant strains were confirmed to be synergistic by the fractional inhibitory concentration index . These results demonstrate that for some azole-resistant C . albicans strains, LF-related compounds combined with triazoles can inhibit the growth of hyphae, an important form of this organism in pathogenesis.

Analyst, 1998 Mar, 123(3), 503 - 7
Effects of copper and zinc ions on the germicidal properties of two popular pharmaceutical antiseptic agents cetylpyridinium chloride and povidone-iodine; Zeelie JJ et al.; The effects of copper and zinc ions on the rate of killing of Gram-negative bacterium Pseudomonas aeruginosa, Gram-positive bacterium Staphylococcus aureus and fungal yeast Candida albicans by antiseptic agents cetylpyridinium chloride and povidone-iodine (Betadine) were investigated . In the 48 test cases copper and zinc ions clearly potentiated the antiseptic agents in 28 (58.3%) cases and exhibited an improved (not clear potentiation) activity in 15 (31.3%) cases . In five (10.4%) cases there was no change in the antiseptics' antimicrobial activity . In general zinc potentiated the antiseptic agents more than copper . If an 'improved activity' was the only criterion for this study, then a more rapid antimicrobial effect was observed in 43 out of the 48 test cases, i.e., 90%.

Injury, 1998 Jan, 29(1), 35 - 40
Intact autocrine activation and cytokine production by PMNs from injured adults with elevated Candida antigen titres; Sweeney JF et al.; Injured patients with Candida antigen titres have increased mortality due to sepsis . Polymorphonuclear leucocytes (PMNs) from injured patients with elevated Candida antigen titres demonstrate impaired function against Candida albicans growth when compared with PMNs from injury matched controls . To determine if PMN dysfunction is global, PMNs from patients with positive Candida antigen titres were evaluated for their ability to activate the anticandidal function of normal PMNs (autocrine activation) and to produce tumour necrosis factor (TNF) and interleukin 8 (IL8), known activators of PMN anticandidal function, this study demonstrates that the PMN dysfunction is not global, as PMN cytokine production and autocrine activation remain intact.

J Bacteriol, 1998 Jul, 180(14), 3657 - 62
Tca1, the retrotransposon-like element of Candida albicans, is a degenerate and inactive element; Chen J et al.; Candida albicans is an asexual fungus and as such must rely on mechanisms other than sexual recombination to generate genetic diversity . Retrotransposons are ubiquitous genetic elements known to generate multiple types of genomic alterations . We have further investigated the nature of the retrotransposon-like element Tca1 in C . albicans . Tca1 is present at two loci in strain SC5314 . Both loci have now been cloned, and one element was sequenced in its entirety . This element was flanked by alpha elements, or long terminal repeats (LTRs), and contained an intervening region of 5,614 bp . The intervening region was highly degenerate and contained no extended open reading frames, indicating that Tca1 is not a functional element . Partial sequence determination demonstrated that the elements from the two loci were nearly identical . Genetic manipulation of the elements showed that both loci were heterozygous for Tca1, that both were transcriptionally active, and that deletion of both had no effect on growth rate or germ tube formation . Thus, it is unclear why this nonfunctional, highly degenerate element has been maintained in many clinical isolates.

Gerontology, 1998, 44(4), 192 - 7
Influence of age on oxidative bursts (chemiluminescence) of polymorphonuclear neutrophil leukocytes; Braga PC et al.; The release of reactive oxygen species (ROS) during neutrophil oxidative bursts is the last of a sequence of different steps leading to the neutralization of pathogen microorganisms . Using luminol-amplified chemiluminescence (LACL), the oxidative burst activity of neutrophils in elderly people (> or = 75 years) was compared with that in younger controls (39 years on average) after activation with both particulate (Candida albicans) and soluble (formyl-methionyl-leucyl-phenylalanine; fMLP) stimulants . After Candida stimulation, a reduction in LACL was observed in the elderly subjects in comparison with the controls, but the difference did not reach statistical significance . After fMLP stimulation, the reduction in LACL was significant, thus suggesting that the Candida pathway of chemiluminescence production seems to be less affected than the fMLP pathway . This finding raises questions concerning the complex differences in the pathways of cell killing and ROS generation, and their efficacy in the elderly . Various possible explanations are discussed, all of which need further investigation.

Biochim Biophys Acta, 1998 Jul 9, 1398(3), 359 - 64
Molecular cloning of a gene encoding phospholipase D from the pathogenic and dimorphic fungus, Candida albicans; Kanoh H et al.; A phospholipase D gene (CaPLD) has been cloned from the Candida albicans genomic DNA library . The CaPLD is a member of a highly conserved gene family of PLD and has the highest homology to Saccharomyces cerevisiae PLD (SPO14) with an overall homology of 42% . Phylogenetic analysis indicated that fungus PLDs including CaPLD composed one of the three clusters of PLD genes.

Pediatr Infect Dis J, 1998 Jun, 17(6), 504 - 8
Candidemia in a neonatal intensive care unit: trends during fifteen years and clinical features of 111 cases; Kossoff EH et al.; OBJECTIVE: To determine changes in the incidence of candidemia in a neonatal intensive care unit (NICU) during a 15-year period (1981 to 1995) and to compare the prevalence and case fatality rates of Candida albicans and Candida parapsilosis infections . METHODS: A retrospective study was conducted of candidemia occurring in infants in a NICU between January 1, 1981, and December 31, 1995 . Cases were identified through computerized searching of a microbiology blood culture database . Candidemia was considered contributory to mortality if death occurred within 3 days of positive blood cultures or if there was autopsy evidence of disseminated candidiasis . RESULTS: One hundred eleven cases of candidemia occurred in 107 infants, representing 1% of all NICU patients during the study period . The rate of candidemia in the NICU increased from 2.5 cases per 1000 admissions in 1981 to 1985, to 4.6 per 1000 admissions in 1986 to 1990 and to 28.5 per 1000 in 1991 to 1995 (P = 0.001) . C . albicans was the predominant cause of candidemia between 1981 and 1990 . C . parapsilosis was the most prevalent species between 1991 and 1995, causing 53 of 89 cases (60%) . The mortality from C . albicans, 13 of 50 cases (26%), was significantly higher than the mortality from C . parapsilosis, 2 of 54 (4%) (P = 0.002; relative risk, 7; 95% confidence interval, 1.7 to 30) . CONCLUSIONS: The rate of candidemia in our neonatal intensive care unit increased >11-fold in the 15 years from 1981 to 1995; the prevalent Candida species shifted from C . albicans to C . parapsilosis; and candidemia associated with C . albicans has significantly higher mortality than with C . parapsilosis.

J Infect Dis, 1998 Jul, 178(1), 227 - 34
Genetic basis for protection against experimental vaginal candidiasis by peripheral immunization; Mulero-Marchese RD et al.; In these studies, significant protection against experimental vaginal candidiasis after a subcutaneous immunization with Candida albicans extract was achieved in BALB/c mice but not in C57BL/6 (B6) mice . Protection from vaginal candidiasis was transferred to naive BALB/c mice by a population of spleen cells derived from immunized BALB/c mice . Removal of CD3 or CD4 but not CD8 T cells before transfer completely abrogated resistance to vaginal candidiasis . Recombinant inbred (RI) strains of mice derived from BALB/c and B6 strains were used for mapping loci that might be responsible for regulating vaginal protection after subcutaneous immunization . Linkage analysis using microsatellite-based genome mapping in these RI strains revealed four candidate loci on chromosomes 3, 7, 8, and 18 that exhibit statistically significant linkage to the strain distribution pattern . These results may contribute to the understanding of host genetic factors controlling the immune response to vaginal infections.

J Clin Microbiol, 1998 Jul, 36(7), 2093 - 5
Simple, inexpensive, reliable method for differentiation of Candida dubliniensis from Candida albicans; Pinjon E et al.; Candida dubliniensis is a recently described pathogenic species which shares many phenotypic features with Candida albicans, including the ability to form germ tubes and chlamydospores . These similarities have caused significant problems in the identification of C . dubliniensis by the average clinical mycology laboratory . To facilitate the differentiation of these species, we investigated the growth of 120 isolates of C . dubliniensis and 98 C . albicans isolates at 42 and 45 degrees C on Emmons' modified Sabouraud glucose agar (SGA) and 10 isolates of each species in yeast-peptone-dextrose broth . None of the C . dubliniensis isolates grew on the agar or in the broth medium at 45 degrees C, while 11 isolates were capable of growing on SGA at 42 degrees C . In contrast, all of the C . albicans isolates but one grew at 45 degrees C on or in either medium . These reproducible results clearly demonstrate that the incubation of isolates suspected to be C . dubliniensis or C . albicans at 45 degrees C provides a simple, reliable, and inexpensive method for the differentiation of the two species.

J Clin Microbiol, 1998 Jul, 36(7), 1886 - 9
International surveillance of bloodstream infections due to Candida species: frequency of occurrence and antifungal susceptibilities of isolates collected in 1997 in the United States, Canada, and South America for the SENTRY Program . The SENTRY Participant Group; Pfaller MA et al.; An international program of surveillance of bloodstream infections (BSIs) in the United States, Canada, and South America between January and December 1997 detected 306 episodes of candidemia in 34 medical centers (22 in the United States, 6 in Canada, and 6 in South America) . Eighty percent of the BSIs were nosocomial and 50% occurred in patients hospitalized in an intensive care unit . Overall, 53.3% of the BSIs were due to Candida albicans, 15.7% were due to C . parapsilosis, 15.0% were due to C . glabrata, 7.8% were due to C . tropicalis, 2.0% were due to C . krusei, 0.7% were due to C . guilliermondii, and 5.8% were due to Candida spp . However, the distribution of species varied markedly by country . In the United States, 43.8% of BSIs were due to non-C . albicans species . C . glabrata was the most common non-C . albicans species in the United States . The proportion of non-C . albicans BSIs was slightly higher in Canada (47.5%), where C . parapsilosis, not C . glabrata, was the most common non-C . albicans species . C . albicans accounted for 40.5% of all BSIs in South America, followed by C . parapsilosis (38.1%) and C . tropicalis (11.9%) . Only one BSI due to C . glabrata was observed in South American hospitals . Among the different species of Candida, resistance to fluconazole (MIC, > or = 64 microg/ml) and itraconazole (MIC, > or = 1.0 microg/ml) was observed with C . glabrata and C . krusei and was observed more rarely among other species . Isolates of C . albicans, C . parapsilosis, C . tropicalis, and C . guilliermondii were all highly susceptible to both fluconazole (99.4 to 100% susceptibility) and itraconazole (95.8 to 100% susceptibility) . In contrast, 8.7% of C . glabrata isolates (MIC at which 90% of isolates are inhibited {MIC90}, 32 microg/ml) and 100% of C . krusei isolates were resistant to fluconazole, and 36.9% of C . glabrata isolates (MIC90, 2.0 microg/ml) and 66.6% of C . krusei isolates were resistant to itraconazole . Within each species there were no geographic differences in susceptibility to fluconazole or itraconazole.

Clin Exp Immunol, 1998 May, 112(2), 334 - 40
Effects of granulocyte colony-stimulating factor (G-CSF) treatment on granulocyte function and receptor expression in patients with ventilator-dependent pneumonia; Hustinx WN et al.; Considerable experimental evidence in animals suggests that treatment with G-CSF may have a beneficial effect in the management of severe infections in non-neutropenic hosts . This beneficial effect is attributed to an enhancement of granulopoiesis and neutrophil function, the latter possibly involving up-regulation of receptors on neutrophils that are involved in antibody-mediated cytotoxicity and killing of microorganisms . We compared neutrophil function and phenotype in blood and bronchoalveolar lavage fluid (BALF) of 10 patients with severe ventilator-dependent pneumonia, at baseline and following initiation of G-CSF treatment as adjunct to standard therapy . G-CSF treatment was associated with three-fold increased blood neutrophil counts at day 3 of treatment compared with baseline counts . Mean serum G-CSF concentration increased from 313 to 2007 pg/ml . After correction for lavage dilution effects, BALF G-CSF levels did not differ significantly from baseline, nor did neutrophil receptor expression (FcgammaRI, FcgammaRII, FcgammaRIII, CR3, and L-selectin) or indicators of neutrophil function such as respiratory burst activity, phagocytosis and killing of Candida albicans in BALF or blood . The mortality in this group of patients was 30% and compared favourably to the APACHE II-derived predicted mortality of 60% . We conclude that the possible therapeutic benefit of G-CSF administration in the early phase of severe bacterial pneumonia is not readily explained by its effect on baseline indicators of neutrophil function or receptor expression.

Biosci Biotechnol Biochem, 1998 May, 62(5), 1025 - 7
Insecticidal and antifungal activities of aminorhodanine derivatives; Inamori Y et al.; Aminorhodanine (1) showed strong insecticidal activity against Culex pipiens pallens and Musca domestica, with respective LD50 values of 0.21 microgram/insect and 0.87 microgram/insect . Compound 1 had antifungal activity against Aspergillus niger ATCC-16404, Trichophyton mentagrophytes IFO-32412, Candida albicans ATCC-10231, Hansenula anomala OPS-308 and Penicillium expansum IFO-8800 . In particular, 1 had potent antifungal activity against Aspergillus niger ATCC-16404, its minimal inhibitory concentration (MIC) being 6.25 micrograms/ml . Both activities of 1 were much higher than those of rhodanine (4), suggesting that the introduction of an amino group into N-3 of 4 plays an important role in the biological activity of rhodanine-related compounds . On the other hand, N-acetylaminorhodanine (2) and N-benzoylaminorhodanine (3) did not show either activity, suggesting that the free amino group at N-3 of 1 is closely related to the inhibitory activity of rhodanine derivatives.

J Endocrinol Invest, 1998 May, 21(5), 329 - 33
Cushing's syndrome complicated by multiple opportunistic infections; Bakker RC et al.; The case history of a 56-year-old man is described who suffered from severe adrenocorticotrophic hormone (ACTH)-dependent Cushing's syndrome . The clinical course was complicated by simultaneous infections with Pneumocystis carinii, Staphylococcus aureus, Candida albicans, Aspergillus fumigatus and Herpes simplex, which proved to be fatal . A study of the literature shows that opportunistic infections in endogenous Cushing's syndrome are associated with severe cortisol excess and carry a high mortality . Opportunistic infections are most prevalent in the ectopic ACTH syndrome explained by the very high plasma cortisol concentrations in this condition . Infections with Aspergillus species, Cryptoccus neoformans, Pneumocystis carinii and Nocardia asteroides predominated . Cushing's syndrome with a very high plasma cortisol concentration causes a severe immunocompromized state . Prompt evaluation of the cause of the hypercortisolism, initiation of cortisol lowering therapy, primary prophylaxis for Pneumocystis carinii infection when plasma cortisol exceeds 2500 nmol L-1 and a search for concomitant infectious disease is recommended.

Rocz Akad Med Bialymst, 1997, 42 Suppl 2, 208 - 11
Ultrastructural changes of Candida albicans under influence of cyclosporin A; Krajewska-Kulak E et al.; The aim of this study was the assessment of influence of cyclosporin A on ultrastructural changes of Candida albicans in vitro . Strains of Candida albicans isolated from woman with chronic vulvovaginitis were used . At the first part of experiment MIC (minimal inhibitory concentration) of cyclosporin on Sabouraud medium was assessed . Control plates were also prepared . From plates with Candida albicans there was observed its visual growth, the yeasts were prepared for electron transmission microscopic examines . It was found that ultrastructural changes of Candida albicans cells in the presence of Cyclosporin A . In particular, the cell wall was thicker and deteriorated . The organelles were altered in varying degrees and autolysis was observed.

Mycopathologia, 1997-98, 140(2), 89 - 97
Experimental candidal mastitis in goats: clinical, haematological, biochemical and sequential pathological studies; Singh P et al.; The present study, first of its kind, was conducted with the objectives to understand hitherto little known aspects of candidal mastitis, like its sequential pathology, pathogenesis and clinico-biochemical changes . For this purpose, unilateral intramammary inoculation of 10 goats with Candida albicans (1.2 x 10(7) yeast cells) resulted in the development of mastitis, with gross and microscopic lesions being restricted to the infected udder halves only and without dissemination of infection to the opposite uninfected udder halves as well as other organs of the body . The experiment was continued for 40 days and after infection, there was sharp fall in milk yield and Candida albicans was directly demonstrated in the milk and re-isolated from the milk and udder tissues up to 30th day after inoculation . An increase in total immunoglobulins in the milk and plasma along with increase in total plasma proteins were also observed . Haematology revealed leukocytosis and neutrophilia . Microscopically, there was acute purulent mastitis, which later became chronic, nonpurulent and interstitial with formation of granulomas . It was concluded that Candida albicans was highly pathogenic to the lactating goat mammary gland even without immunosuppression or antibiotic treatment, resulting in severe irreversible tissue damage and nearly complete agalactia.

Mycopathologia, 1997-98, 140(2), 85 - 7
Prevalence of otomycosis in malnourished children in Edo State, Nigeria; Enweani IB et al.; Out of the total number at 200 suspected cases of otomycoses consisting of 40 malnourished and 160 apparently healthy children examined in this study between the months of July and August in Edo State, 64 Cases (32%) were identified to be of fungal aetiology on the basis of positive culture and careful microscopic examination . The state at protein energy malnourishment was deterwined using physicians' comments in their case files . The fungal agents isolated were Aspergillus niger 28 (43.8%); A . fumigatus 4 (25%); Fusarium solari 4 (6.3%); Candida albicans 8 (12.5%); and Hendersonula teruloidea types torn B 5 (6.3%) . Of these isolates, A . niger having an solation rate of (43.8%) was found to be the most predominant fungal species associated with otomycosis.

Mycopathologia, 1997-98, 140(2), 69 - 76
Effect of sub-inhibitory concentration of chlorhexidine on lipid and sterol composition of Candida albicans; Abu-Elteen KH et al.; The effect of a sub-inhibitory concentration of chlorhexidine on lipid and sterol composition of Candida albicans was investigated . The total lipid content of this yeast grown in the presence of chlorhexidine was reduced whilst the total sterol content was increased compared with control-grown cells . Lipids and sterol analyses of this yeast grown in the presence and absence of chlorhexidine are presented . Chlorhexidine-grown yeast had a higher level of phosphatidylethanolamine, phosphatidylcholine and monogalactosyldiacylglycerol . Lower proportions of phosphatidylinositol plus phosphatidylserine, phosphatidic acid and cardiolipin were found in C . albicans grown in the presence of the drug when compared with control-grown yeast . The major fatty acids in control-grown cells were C16 and C18 . Drug grown-cells had higher proportions of palmitic acid (16:0) and stearic acid (18:0), but lower proportions of palmitoleic acid (16:1) and oleic acid (18:1) . Chlorhexidine also decreased the unsaturated-to-saturated fatty acid ratio, while the C16/C18 ratios increased compared to control-grown cells . Differences in the fatty acid composition of major phospholipids and neutral lipids between drug and control-grown yeast were also detected . Sterol analysis of control-grown cells showed that the major sterol present was ergosterol (55.4% wt) . A significant increase in ergosterol and obtusifoliol was observed in chlorhexidine-treated cells and a significant decrease in squalene and lanosterol . Our results suggested that chlorhexidine affected the lipid and sterol composition of C . albicans.

Am J Ophthalmol, 1998 Jun, 125(6), 873 - 5
Endogenous Candida endophthalmitis after induced abortion; Chen SJ et al.; PURPOSE: To report two young healthy women who developed endogenous Candida endophthalmitis after undergoing surgically induced abortion . METHOD: Case reports . RESULTS: In two eyes of two patients, a diagnosis of Candida endophthalmitis was established by typical fundus appearance, positive vaginal culture, and, in one case, positive vitreous culture . After vitrectomy and intravitreal amphotericin B injection, one eye of one patient had a best-corrected visual acuity of 20/200, whereas one eye of one patient, who had systemic corticosteroid treatment before the correct diagnosis, developed recurrent retinal detachment and a best-corrected visual acuity of counting fingers . CONCLUSIONS: Induced abortion may cause endogenous Candida endophthalmitis in young healthy pregnant women . Systemic corticosteroid treatment may increase the risk of endophthalmitisPIP: Reported, in this article, are the cases of two young women who developed endogenous Candida endophthalmitis after induced abortion . Both women experienced transient fever, chills, and abdominal pain after the abortion and were given antibiotics . The diagnosis of endophthalmitis was established on the basis of typical fundus appearance, positive vaginal culture, and (in one case) positive vitreous culture . In the first woman, who received vitrectomy and intravitreal amphotericin B injection, the affected eye had a best corrected visual acuity of 20/200 . In the second woman, who was given systemic corticosteroid treatment before the correct diagnosis was reached, recurrent retinal detachment developed and the best corrected visual acuity was counting fingers . It appears that Candida organisms harbored in the genital tract are directly inoculated into the venous system during induced abortion . Once in the blood, if sufficient fungal load is present, Candida albicans tends to localize in the choroid and to spread toward the retina and vitreous cavity . The immunosuppressive effect of corticosteroids further increases the risk of endophthalmitis .

Curr Genet, 1998 Jun, 33(6), 451 - 9
Candida albicans ALS3 and insights into the nature of the ALS gene family; Hoyer LL et al.; The ALS1 (agglutinin-like sequence) gene of Candida albicans encodes a protein similar to alpha-agglutinin, a cell-surface adhesion glycoprotein of Saccharomyces cerevisiae (Hoyer et al . 1995) . A central domain of a tandemly repeated 108-bp sequence is found in the ALS1 coding region . This tandem-repeat motif hybridizes to multiple C . albicans genomic DNA fragments, indicating the possibility of other ALS1-like genes in C . albicans (Hoyer et al . 1995) . To determine if these fragments constitute a gene family, tandem-repeat-hybridizing genomic fragments were isolated from a fosmid library by PCR screening using primers based on the consensus tandem-repeat sequence of ALS1 (Hoyer et al . 1995) . One group of fosmids, designated ALS3, encodes a gene with 81% identity to ALS1 . The sequences of ALS1 and ALS3 are most conserved in the tandem-repeat domain and in the region 5' of the tandem repeats . Northern-blot analysis using unique probes from the 3' end of each gene demonstrated that ALS1 expression varies, depending on which C . albicans strain is examined, and that ALS3 is hyphal-specific . Both genes are found in a variety of C . albicans and C . stellatoidea strains examined . The predicted Als1p and Als3p exhibit features suggesting that both are cell-surface glycoproteins . Southern blots probed with conserved sequences from the region 5' of the tandem repeats suggest that other ALS-like sequences are present in the C . albicans genome and that the ALS family may be larger than originally estimated.

Quintessence Int, 1998 Feb, 29(2), 107 - 13
A comparison of three methods for detecting Candida albicans in patients with Sjögren's syndrome; Rhodus NL et al.; OBJECTIVE: An immediate chairside technique (Latex Candida) for the detection of Candida albicans was compared with a simple tube culturing technique (Oricult) and the traditional laboratory culturing technique in patients with Sjogren's syndrome . METHOD AND MATERIALS: Subjects with primary (n = 9) and secondary (n = 9) Sjogren's syndrome (mean age of 56.7 years; all female) and an age- and sex-matched group of control subjects (n = 9) were selected . Three different methods for culturing Candida albicans were performed for each subject . One culturette was plated on a trypticase soy-agar streptomycin-vancomycin medium plate and incubated for 48 hours at 37 degrees C . Another swab was plated on a reagent paper with the Latex Candida test kit . The third swab was placed in a culture media tube using the Oricult kit and incubated for 48 hours at 37 degrees C . RESULTS: All three techniques indicated a significant difference in the prevalence of Candida between the control group and both Sjogren's groups . The Latex Candida technique indicated that 78% of all Sjogren's subjects were positive for Candida, while the other two tests indicated that 83% were positive . CONCLUSION: The Latex Candida technique was comparable to Oricult and streptomycin-vancomycin culturing techniques for negative results and was correctly positive for 90% of cases.

Farmaco, 1998 Mar, 53(3), 224 - 32
Synthesis, antimicrobial activity and bleaching effect of some reaction products of 4-oxo-4H-benzopyran-3-carboxaldehydes with aminobenzothiazoles and hydrazides; el-Shaaer HM et al.; The synthesis of the biologically active novel systems derived from reaction of 3-formylchromones with three types of amino derivatives, 6-R2-2-aminobenzothiazoles, 6-amino-2-R3-thiobenzothiazoles and hydrazide derivatives (derived from cyanoacetic, isonicotine, salicylic and gallic acids) was carried out . The structures of the prepared compounds have been proved by elemental analysis, 1H NMR and IR spectra . Antimicrobial activity was studied against the following microorganisms--bacteria G+ (Staphylococcus aureus 29/58, Bacillus subtilis 18/66), G- (Escherichia coli 326/71, Pseudomonas aeruginosa); yeasts: Candida albicans, Saccharomyces cerevisiae; moulds: Microsporum gypseum, Aspergillus niger, Scopulariopsis brevicaulis; and against typical and atypical mycobacteria: Mycobacterium tuberculosis (H37Rv), Mycobacterium kansasii (PFG 8), Mycobacterium avium (My 80/72), Mycobacterium fortuitum (1021) . The hereditary bleaching effect on the plastid system of Euglena gracilis, a unique phenomenon of the biological activity of chromone derivatives, is reported . The bleaching test on E . gracilis is used for detecting extranuclear mutations.

Wei Sheng Wu Xue Bao, 1996 Jun, 36(3), 161 - 7
{A retrotransposon-like element Tca1 was used for taxonomic determination of Candida albicans}; Chen J et al.; We had isolated from Candida albicans a moderately repetitive sequence designated alpha and a retrovirus-like transposable element Tcal (Transposon Candida albicans) . The Tcal consisted of two 388bp direct repeats of the alpha element, called LTR (Long Termination Repeat), which was separated by approximately 5.5kb of DNA . A large number of strains from America and China have been grouped based on patterns of hybridization bands visualized on Southern blots of EcoRI digested genomic DNA probed with alpha and Tcal element internal sequence . Strains from same area have higher relatedness than those from different area . The hybridization patterns with URA3 and other DNA probes were also conserved within the groups . alpha element are species specific, no hybridization was observed with genomic DNA of other yeast species . The data presented here indicate that the alpha element can be employed to distinguish between species and to assess strain relatedness within C . albicans, we suggest that Tcal may be relevant to the genomic evolutions of C . albicans and the pathogenic potential of the organism.

Yeast, 1998 May, 14(7), 681 - 6
Genetic organization and sequence analysis of the hypha-specific cell wall protein gene HWP1 of Candida albicans; Staab JF et al.; A previously isolated partial cDNA encoding a cell wall protein antigen found on hyphal surfaces of the opportunistic fungal pathogen, Candida albicans (Staab et al., 1996) was used to clone the complete hyphal wall protein 1 gene (HWP1) . Hyphal forms of C . albicans invade mucosal surfaces of immunocompromised patients such as those with AIDS . HWP1 consisted of an open reading frame predicting an acidic protein (pI of 3.37) with a calculated molecular size of 61,122 . The antigenic domain was located in the N-terminal third of the protein . The remainder of the protein contained abundant hydroxy amino acids, and terminated with a string of 15 amino acids typical of sequences specifying post-translational modification with glycosylphosphatidylinositol (6PI) . The analyses suggested that Hwp1 is a glucan-linked protein with serine/threonine-rich regions that are predicted to function in extending a ligand-binding domain into the extracellular space.

Yeast, 1998 May, 14(7), 675 - 80
Isolation and characterization of the Candida albicans SEC4 gene; Clement M et al.; The SEC4 gene product is a major component of the protein secretion machinery . More specifically, it is believed to play a pivotal role in targeting and fusion of secretory vesicles to the plasma membrane . Its recently described implication with the Saccharomyces cerevisiae Rho3p, which is required for directing growing points during bud formation, has prompted us to investigate the role and function of Sec4p in the morphological changes of the yeast pathogen Candida albicans . We have therefore cloned the C . albicans SEC4 gene . It encodes a 210 amino acids long protein sharing up to 75% homology to the S . cerevisiae homolog, when conserved changes are allowed . Its RNA is constitutively expressed in C . albicans grown under various physiological conditions . We also show that it can functionally complement a S . cerevisiae sec4 thermosensitive mutant.

Yeast, 1998 May, 14(7), 665 - 74
Identification of a putative histidine kinase two-component phosphorelay gene (CaHK1) in Candida albicans; Calera JA et al.; We have cloned and analysed the sequence of a putative histidine kinase, two-component gene (CaHK1) from Candida albicans . This gene encodes a 2471 amino acid protein (Cahk1p) with an estimated molecular mass of 281.8 kDa . A homology search of Cahk1p with other proteins in the databases showed that Cahk1p exhibits the greatest homology at its C-terminus with both the sensor and regulator components of prokaryotic and eukaryotic two-component histidine kinases . A further analysis of this homology showed that the Cahk1p possessed both sensor and regulator domains in the same polypeptide . Also, Cahk1p is likely to be a soluble protein . The sensor kinase domain of Cahk1p contains conserved motifs that are characteristic of all histidine kinase proteins, including the putative histidine which is believed to be autophosphorylated during activation, ATP binding motifs and others (F- and N-motifs), with unknown function . The Cahk1p regulator domain also contains conserved aspartate and lysine residues and the putative aspartate, which is secondarily phosphorylated by the autophosphorylated histidine . Finally, according to the codon usage frequency of the CaHK1 gene in comparison with other genes from C . albicans, there would appear to be a low level of expression of the gene.

Zentralbl Bakteriol, 1998 May, 287(4), 501 - 8
Characterization of pathogenetic determinants of Candida albicans strains; Macura AB et al.; The study was an attempt to correlate phenotypic pathogenetic determinants of clinical Candida albicans strains with their genotype as determined by PCR fingerprinting . A total of 25 C . albicans strains was tested . Adherence capacity, hydrophobicity and proteinase production were compared with the genotypes of the particular Candida strains . The fungal strains represented eleven genotypes . No correspondence relationship was found between genotype and the markers of pathogenicity.

Cell Immunol, 1998 May 25, 186(1), 28 - 38
Interaction between human interleukin-2-activated natural killer cells and heat-killed germ tube forms of Candida albicans; Arancia G et al.; Human interleukin-2-activated natural killer (LAK) cells are able to recognize and to bind to both live and heat-killed germ tube forms of Candida albicans, establishing a wide and intimate contact as revealed by electron microscopic observations . Following the interaction, LAK cells are activated: an increased expression of some cytokine mRNA (in particular, TNF-alpha, GM-CSF, and IFN-gamma) has been revealed by RT-PCR and perforin secretion has been suggested by immunofluorescence microscopy . Nonetheless, neither morphological damage or growth inhibition of fungal target cells have been detected . Instead, evident signs of cell damage could be noticed in interacting LAK cells . Moreover, the observation by transmission electron microscopy of LAK cell-germ tube conjugates revealed the presence of apoptotic cells . The analysis of LAK cell cytotoxic activity against DAUDI cells showed that the lymphocytic effector underwent a significant reduction in its lytic capability after the interaction with C . albicans . The results obtained in this in vitro study seem to indicate that in such an interaction LAK cells cannot directly inhibit or kill the fungal pathogen by using their lytic machinery but they secrete those cytokines which have stimulatory effects on phagocytic cells . The ultimate results are the programmed death of LAK cells and the enhancement of the fungicidal activity exerted by competent cells.

Cell Immunol . 1998 Apr 10;185(1):82.
Papers to Appear in Forthcoming Issues; The pH of the host niche controls gene expression in and virulence of Candida albicans; Laboratory of Bacteriology and Medical Mycology, Instituto Superiore di Sanita, 00161 Rome, ItalyLittle is known of the biological attributes conferring pathogenicity on the opportunistic fungal pathogen Candida albicans . Infection by this pathogen, as for bacterial pathogens, may rely upon environmental signals within the host niche to regulate the expression of virulence determinants . To determine if C . albicans responds to the pH of the host niche, we tested the virulence of strains with mutations in either of two pH-regulated genes, PHR1 and PHR2 . In vitro, PHR1 is expressed when the ambient pH is at 5.5 or higher and deletion of the gene results in growth and morphological defects at neutral to alkaline pHs . Conversely, PHR2 is expressed at an ambient pH below 5.5, and the growth and morphology of the null mutant is compromised below this pH . A PHR1 null mutant was avirulent in a mouse model of systemic infection but uncompromised in its ability to cause vaginal infection in rats . Since systemic pH is near neutrality and vaginal pH is around 4.5, the virulence phenotype paralleled the pH dependence of the in vitro phenotypes . The virulence phenotype of a PHR2 null mutant was the inverse . The mutant was virulent in a systemic-infection model but avirulent in a vaginal-infection model . Heterozygous mutants exhibited partial reductions in their pathogenic potential, suggesting a gene dosage effect . Unexpectedly, deletion of PHR2 did not prevent hyphal development in vaginal tissue, suggesting that it is not essential for hyphal development in this host niche . The results suggest that the pH of the infection site regulates the expression of genes essential to survival within that niche . This implies that the study of environmentally regulated genes may provide a rationale for understanding the pathobiology of C . albicans.

APMIS, 1998 Apr, 106(4), 441 - 8
Adherence of Staphylococcus aureus, Klebsiella pneumoniae and Candida albicans to human buccal epithelial cells, from healthy persons and HIV carriers, under the influence of Broncho Vaxom in vitro and ascorbic acid in vivo; Fortis AA et al.; We examined the in vitro effect of Broncho Vaxom (BV) (an immunobiotherapeutic preparation containing a lysate made from bacteria often involved in respiratory tract infections) on adherence of Staphylococcus aureus, Klebsiella pneumoniae and Candida albicans to human buccal epithelial cells (BEC) of healthy volunteers and HIV carriers . We also examined the ex vivo effect of ascorbic acid on the adherence of the same microorganisms to BEC of HIV carriers . The study reached the following conclusions: The presence of BV in vitro significantly reduces the adherence of the tested strains to BEC from healthy persons and HIV carriers . No significant difference was observed between healthy persons and HIV carriers regarding the adherence of the tested strains to BEC . Significant difference in the adherence of the tested strains to BEC was observed between HIV carriers who had been taking ascorbic acid over a 3-month period and those who had not . There was no further reduction in the adherence of the tested strains to BEC from HIV carriers who had been taking ascorbic acid in the presence of BV in vitro.

Clin Infect Dis, 1998 Jun, 26(6), 1330 - 4
Comparison of two methods for the assessment of delayed-type hypersensitivity skin responses in patients with human immunodeficiency virus infection; Martinez-Marcos FJ et al.; We compared two techniques for detecting delayed-type hypersensitivity (DTH) skin responses in 359 patients infected with human immunodeficiency virus (HIV) (mean CD4+ lymphocyte count, 387/microL) . DTH responses were assessed with use of two antigenic panels administered simultaneously: tuberculin purified protein derivative (PPD) plus three control antigens (Candida albicans, mumps antigen, and tetanus toxoid) administered by the Mantoux method and by a multiple-puncture device delivering seven antigens percutaneously (MULTITEST CMI; Institut Merieux, Lyon, France) . Eighty-three patients (23%) were anergic, 216 (60%) reacted to both panels, 55 (15%) did not react to MULTITEST CMI but did react to the antigens administered by Mantoux method, and only five (1%) reacted to MULTITEST CMI without reacting to antigens administered by the Mantoux method (P < .001, McNemar's test) . Each of the three possible combinations of PPD plus two control antigens administered by the Mantoux method were also superior to MULTITEST CMI for classifying patients as nonanergic (P < .001, McNemar's test) . We conclude that the application of antigens by the Mantoux method is more efficient than MULTITEST CMI for detecting DTH skin responses in HIV-infected patients.

Proc Natl Acad Sci U S A, 1998 Jun 23, 95(13), 7670 - 5
Molecular analysis of CaMnt1p, a mannosyl transferase important for adhesion and virulence of Candida albicans; Buurman ET et al.; There is an immediate need for identification of new antifungal targets in opportunistic pathogenic fungi like Candida albicans . In the past, efforts have focused on synthesis of chitin and glucan, which confer mechanical strength and rigidity upon the cell wall . This paper describes the molecular analysis of CaMNT1, a gene involved in synthesis of mannoproteins, the third major class of macromolecule found in the cell wall . CaMNT1 encodes an alpha-1, 2-mannosyl transferase, which adds the second mannose residue in a tri-mannose oligosaccharide structure which represents O-linked mannan in C . albicans . The deduced amino acid sequence suggests that CaMnt1p is a type II membrane protein residing in a medial Golgi compartment . The absence of CaMnt1p reduced the ability of C . albicans cells to adhere to each other, to human buccal epithelial cells, and to rat vaginal epithelial cells . Both heterozygous and homozygous Camnt1 null mutants of C . albicans showed strong attenuation of virulence in guinea pig and mouse models of systemic candidosis, which, in guinea pigs, could be attributed to a decreased ability to reach and/or adhere internal organs . Therefore, correct CaMnt1p-mediated O-linked mannosylation of proteins is critical for adhesion and virulence of C . albicans.

Vopr Med Khim, 1998 Mar-Apr, 44(2), 172 - 8
{Determination of a Candida albicans antigen using an amperometric immunoenzyme sensor}; Kutyreva MP et al.; Determination new variant enzyme immunoassay with amperometric enzyme immunosensor, including the immobilizing enzyme-choline esterase and antibodies against Candida albicans (CA) in biosensitivity part of sensor, for diagnose disease of CA . The method for determination of CA based on combination immunochemical reactions and voltammetric indication of analytical signal was developed . Amperometric enzyme immunosensor developed has been used as detector . Differences dilutions of antibody (Ab) against antigen (Ag) of CA immobilizing in common with choline esterase (CE) . The method of immobilization developed allows to receive the sensor with including the immobilized CE and Ab in common . The method of determination of CA based on combination the reaction of forming immune complex tAb-AgI with enzyme immunosensor for its detection . The dynamic range of concentrations determined of Ag depends on degree of dilution of Ab used for manufactory biosensitivity part of sensor . The data indicate that the {Ab-Ag} immune complexes are stable . This is also confirmed by the values of {Ab-Ag} binding constants, obtained in Scatchard coordinates . This method of determination doesn't require special preparation of a sample . Selectivity, sensitivity, simplicity and quickness are characterize of this method which could be used for manufacturing test-sistem for determination CA in blood.

Aten Primaria, 1998 Apr 15, 21(6), 395 - 8
{Vaginal candidiasis: diagnosis and treatment in a primary care clinic}; Gonzalez-Pedraza Aviles A et al.; OBJECTIVE: To assess the frequency of Candida species others than Candida albicans isolation, in patients with chronic as well as relapsing vaginitis . To analyze the clinical pictures and the treatments routinely employed in patients with Candida albicans infection, compared to those with other Candida species infection, in a Primary Care Clinic . DESIGN: Prospective, comparative, transverse study . SETTING: Primary health center from January, 1995 to May, 1997 . MATERIAL: Two hundred and thirty four women (234) in ages between 16 and 55 years, with cervicovaginitis and a positive culture to any species of Candida, were the subject of the present study . RESULTS: From the total of 234 patients studied, 142 (60.5%) were infected by Candida albicans and 92 (39.5%) by other Candida species; no difference in symptomatology was recorded in both groups . Nistatine alone or combined with imidazole derivatives were the antimycotics more commonly employed, with Candida albicans resistance of 9.6% in the first case and 11.2% in the second one . CONCLUSIONS: It is stressed the importance that has the identification of Candida species causing vaginitis . It is also acknowledged the well handled treatment, despite of the species to be treated.

Clin Microbiol Rev, 1998 Apr, 11(2), 382 - 402
Clinical, cellular, and molecular factors that contribute to antifungal drug resistance; White TC et al.; In the past decade, the frequency of diagnosed fungal infections has risen sharply due to several factors, including the increase in the number of immunosuppressed patients resulting from the AIDS epidemic and treatments during and after organ and bone marrow transplants . Linked with the increase in fungal infections is a recent increase in the frequency with which these infections are recalcitrant to standard antifungal therapy . This review summarizes the factors that contribute to antifungal drug resistance on three levels: (i) clinical factors that result in the inability to successfully treat refractory disease; (ii) cellular factors associated with a resistant fungal strain; and (iii) molecular factors that are ultimately responsible for the resistance phenotype in the cell . Many of the clinical factors that contribute to resistance are associated with the immune status of the patient, with the pharmacology of the drugs, or with the degree or type of fungal infection present . At a cellular level, antifungal drug resistance can be the result of replacement of a susceptible strain with a more resistant strain or species or the alteration of an endogenous strain (by mutation or gene expression) to a resistant phenotype . The molecular mechanisms of resistance that have been identified to date in Candida albicans include overexpression of two types of efflux pumps, overexpression or mutation of the target enzyme, and alteration of other enzymes in the same biosynthetic pathway as the target enzyme . Since the study of antifungal drug resistance is relatively new, other factors that may also contribute to resistance are discussed.

J Antimicrob Chemother, 1998 May, 41(5), 541 - 8
Fluconazole susceptibility and strain variation of Candida albicans isolates from HIV-infected patients with oropharyngeal candidosis; Barchiesi F et al.; Over a 16 month period we conducted a prospective study in a cohort of 45 HIV-positive patients to detect the development of resistance to fluconazole and to analyse the epidemiology of oropharyngeal candidosis (OPC) . Each episode was treated with fluconazole 100 mg/day po for 10 days . All yeast isolates were tested for their in-vitro susceptibility to fluconazole . Multiple strains of Candida albicans simultaneously isolated from a given patient were typed by electrophoretic karyotyping . Overall, 106 episodes of OPC were diagnosed among the 45 patients: 18/45 patients (40%) had only one episode, 11/45 (24%) had two episodes, and the remaining 16/45 (36%) had three or more episodes (range 3-7) . Cure (complete resolution of signs and symptoms and negative post-treatment cultures) and improvement (complete resolution of signs and symptoms but positive post-treatment cultures) were observed in 30/106 (28%) and 69/106 (65%) episodes of OPC, respectively . Failure (absence of improvement or exacerbation of signs and symptoms) was observed in seven episodes (7%) from four patients . In two of these four patients a significant and progressive increase in fluconazole MICs was observed: from 0.25 to 16 mg/L in one patient, and from < or = 0.125 to 32 mg/L in the second one . Tests on multiple colonies from individual isolation plates showed that it was not unusual to obtain different fluconazole MICs, indicating that, in order to avoid misleading results, one should perform in-vitro susceptibility testing by using a multiple colony inoculum rather than an inoculum made from a single colony . A total of 213 strains of C . albicans isolated from seven patients who suffered from four or more episodes of OPC through the course of the study were typed by electrophoretic karyotyping . Five individuals (71%) were infected with yeasts with only one DNA type, while the other two patients showed the presence of two or three different DNA types . The simultaneous presence of multiple types was found only in one of the seven subjects . Our data confirm the efficacy of fluconazole 100 mg/day for the treatment of OPC in HIV patients . Isolation of fluconazole-resistant strains of C . albicans with this regimen is rare . The vast majority of HIV patients are infected with a unique strain of C . albicans throughout each episode of infection . A minority of patients, however, can harbour strains of C . albicans with variable patterns of fluconazole susceptibility simultaneously.

Electrophoresis, 1998 May, 19(5), 675 - 8
Preparative isoelectric focusing and preparative electrophoresis of hydrophobic Candida albicans cell wall proteins with in-line transfer to polyvinylidene difluoride membranes for sequencing; Masuoka J et al.; Hydrophobic proteins in the cell wall of the opportunistic fungal pathogen Candida albicans are involved in adhesion of this organism to host tissue and thus play a role in its pathogenicity . The hydrophobic nature of these proteins results in their loss during purification due to adsorption to apparatus surfaces . This problem, combined with their low abundance, has made it problematic to purify the hydrophobic proteins in sufficient quantity for sequencing or biochemical analysis . We describe a system that combines preparative isoelectric focusing with continuous elution preparative electrophoresis . The system provides a two-dimensional protein separation while maintaining protein solubility and minimizing protein loss due to adsorption . In addition, we have added an in-line transfer of electrophoretic fractions directly to polyvinylidene difluoride (PVDF) membranes, which further reduces both exposure to apparatus surfaces and purification time.

Andrologia, 1998, 30 Suppl 1, 55 - 9
Influence of different uropathogenic microorganisms on human sperm motility parameters in an in vitro experiment; Huwe P et al.; The influence of different uropathogenic microorganisms (E . coli, enterococcus, Pseudomonas aeruginosa, Staphylococcus saprophyticus, Candida albicans) on human sperm motility was studied in vitro with a computer-assisted sperm analyser (CASA) . Native ejaculates were prepared with the swim-up technique and adjusted to 22 x 10(6) spermatozoa ml-1 . The sperm suspension was artificially infected with microorganisms in concentrations varying from 2 x 10(3) to 2 x 10(7) . Sperm motility was examined directly after incubation, 2, 4 and 6 h later using the Mika motion analysis, a computer-based, automatic motility analysis . Former results with E . coli (serotype 06) could be confirmed that a significant inhibitory effect on sperm motility was associated with bacterial growth . Experiments with the enterococcus strain and Staphylococcus saprophyticus indicated no significant influence on sperm motility parameters . Tests with Pseudomonas aeruginosa showed a decrease of progressive motility according to time, but not to different bacterial concentrations . A significant inhibitory effect of Candida albicans was only detected in the samples with the initial bacterial concentration of 2 x 10(7) microorganisms ml-1.

Am J Clin Oncol, 1998 Jun, 21(3), 308 - 12
Comparison of neutrophil and monocyte function by microbicidal cell-kill assay in patients with cancer receiving granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, or no cytokine after cytotoxic chemotherapy: a phase II trial; Nemunaitis J et al.; Functional effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) were prospectively measured by harvesting blood samples from 51 oncology patients (21 who were receiving no cytokines, 14 receiving rhGM-CSF, and 16 who were receiving rhG-CSF) just before cytotoxic chemotherapy (baseline) immediately before the last cytokine dose (pre), 2 hours after the last cytokine dose (post), and 48 hours after the pre period (follow-up) . Neutrophils and monocytes were separated and functional effects were measured by comparing cell-kill percentages, as determined by a microbial cell-kill assay against Staphylococcus aureus and Candida albicans . Optimal cell concentrations (2 x 10(6) monocytes/ml; 4 x 10(6) neutrophils/ml) and effector-to-cell ratios (1:50) were initially determined with blood samples harvested from 23 healthy volunteers . Results in oncology patients indicated that rhGM-CSF improved monocyte-killing activity against S . aureus at follow-up, compared with controls (p = 0.0094) and compared with monocytes from rhG-CSF-treated patients at the post period (p = 0.014) . Cell-killing percentage of the rhGM-CSF-treated patients was also enhanced against C . albicans during the post period, compared with controls (p = 0.011) and rhG-CSF-treated patients (p = 0.067) . Neutrophil activity was not altered by either cytokine . In conclusion, monocyte-induced microbial killing was enhanced in oncology patients receiving rhGM-CSF after cytotoxic chemotherapy, compared with patients receiving rhG-CSF or no cytokines . No differences in neutrophil activity were observed between patients receiving either cytokine.

Proc Natl Acad Sci U S A, 1998 Jun 9, 95(12), 7069 - 73
COS1, a two-component histidine kinase that is involved in hyphal development in the opportunistic pathogen Candida albicans; Alex LA et al.; Two-component histidine kinases recently have been found in eukaryotic organisms including fungi, slime molds, and plants . We describe the identification of a gene, COS1, from the opportunistic pathogen Candida albicans by using a PCR-based screening strategy . The sequence of COS1 indicates that it encodes a homolog of the histidine kinase Nik-1 from the filamentous fungus Neurospora crassa . COS1 is also identical to a gene called CaNIK1 identified in C . albicans by low stringency hybridization using CaSLN1 as a probe {Nagahashi, S., Mio, T., Yamada-Okabe, T., Arisawa, M., Bussey, H . & Yamada-Okabe, H . (1998) Microbiol . 44, 425-432} . We assess the function of COS1/CaNIK1 by constructing a diploid deletion mutant . Mutants lacking both copies of COS1 appear normal when grown as yeast cells; however, they exhibit defective hyphal formation when placed on solid agar media, either in response to nutrient deprivation or serum . In constrast to the Deltanik-1 mutant, the Deltacos1/Deltacos1 mutant does not demonstrate deleterious effects when grown in media of high osmolarity; however both Deltanik-1 and Deltacos1/Deltacos1 mutants show defective hyphal formation . Thus, as predicted for Nik-1, Cos1p may be involved in some aspect of hyphal morphogenesis and may play a role in virulence properties of the organism.

Blood, 1998 Jun 15, 91(12), 4652 - 61
The role of tumor necrosis factor alpha in modulating the quantity of peripheral blood-derived, cytokine-driven human dendritic cells and its role in enhancing the quality of dendritic cell function in presenting soluble antigens to CD4+ T cells in vitro; Chen B et al.; Because dendritic cells (DC) are critically involved in both initiating primary and boosting secondary host immune responses, attention has focused on the use of DC in vaccine strategies to enhance reactivity to tumor-associated antigens . We have reported previously the induction of major histocompatibility complex class II-specific T-cell responses after stimulation with tumor antigen-pulsed DC in vitro . The identification of in vitro conditions that would generate large numbers of DC with more potent antigen-presenting cell (APC) capacity would be an important step in the further development of clinical cancer vaccine approaches in humans . We have focused attention on identifying certain exogenous cytokines added to DC cultures that would lead to augmented human DC number and function . DC progenitors from peripheral blood mononuclear cells (PBMC) were enriched by adherence to plastic, and the adherent cells were then cultured in serum-free XVIVO-15 medium (SFM) for 7 days with added granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) . At day 7, cultures contained cells that displayed the typical phenotypic and morphologic characteristics of DC . Importantly, we have found that the further addition of tumor necrosis factor alpha (TNFalpha) at day 7 resulted in a twofold higher yield of DC compared with non-TNFalpha-containing DC cultures at day 14 . Moreover, 14-day cultured DC generated in the presence of TNFalpha (when added at day 7) demonstrated marked enhancement in their capacity to stimulate a primary allogeneic mixed leukocyte reaction (8-fold increase in stimulation index {SI}) as well as to present soluble tetanus toxoid and candida albicans (10- to 100-fold increases in SI) to purified CD4+ T cells . These defined conditions allowed for significantly fewer DC and lower concentrations of soluble antigen to be used for the pulsing of DC to efficiently trigger specific T-cell proliferative responses in vitro . When compared with non-TNFalpha-supplemented cultures, these DC also displayed an increased surface expression of CD83 as well as the costimulatory molecules, CD80 and CD86 . Removal of TNFalpha from the DC cultures after 2 or 4 days reduced its enhancing effect on DC yield, phenotype, and function . Thus, the continuous presence of TNFalpha over a 7-day period was necessary to achieve the maximum enhancing effect observed . Collectively, our findings point out the importance of exogenous TNFalpha added to cultures of cytokine-driven human DC under serum-free conditions, which resulted in an enhanced number and function of these APC . On the basis of these results, we plan to initiate clinical vaccine trials in patients that use tumor-pulsed DC generated under these defined conditions.

Int J Antimicrob Agents, 1998 Apr, 10(1), 83 - 6
Antimicrobial activity of antiseptic-coated orthopaedic devices; Darouiche RO et al.; Antimicrobial coating of medical devices, including fracture fixation devices, has evolved as a potentially effective method for preventing device-related infections . We examined the in vitro antimicrobial activity of titanium cylinders coated with the antiseptic combination of chlorhexidine and chloroxylenol . The coated devices provided zones of inhibition against Staphylococcus epidermidis, S . aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans, at baseline and up to 8 weeks after incubation of the coated cylinders in human serum at 37 degrees C . This durable antimicrobial activity was attributed to the relatively slow leaching of chlorhexidine and chloroxylenol from the coated cylinders as measured by high-performance liquid chromatography . These results suggest that antiseptic-coated orthopaedic devices may provide broad-spectrum and durable antimicrobial protection against device-related infection.

Antimicrob Agents Chemother, 1998 Jun, 42(6), 1424 - 7
Antifungal activity of 3'-deoxyadenosine (cordycepin); Sugar AM et al.; The antifungal activity of the nucleoside analog 3'-deoxyadenosine (cordycepin) was studied in a murine model of invasive candidiasis . When protected from deamination by either deoxycoformycin or coformycin, both of which are adenosine deaminase inhibitors, cordycepin exhibited potent antifungal efficacy, as demonstrated by prolongation of survival and a decrease in CFU in the kidneys of mice treated with cordycepin plus an adenosine deaminase inhibitor . The antifungal effect was seen with three different Candida isolates: Candida albicans 64, a relatively fluconazole-resistant clinical isolate of C . albicans (MIC, 16 micrograms/ml), and the fluconazole-resistant Candida krusei . Cordycepin and related compounds may provide another avenue for the discovery of clinically useful antifungal drugs.

Antimicrob Agents Chemother, 1998 Jun, 42(6), 1382 - 6
Assessment of antifungal activities of fluconazole and amphotericin B administered alone and in combination against Candida albicans by using a dynamic in vitro mycotic infection model; Lewis RE et al.; We evaluated the pharmacodynamic activities of fluconazole and amphotericin B given alone and in combination against Candida albicans by using an in vitro model of bloodstream infection that simulates human serum pharmacokinetic parameters for these antifungals . Fluconazole was administered as a bolus into the model to simulate regimens of 200 mg every 24 h (q24 h) and 400 mg q24 h . Amphotericin B was administered at doses producing the peak concentration (2.4 micrograms/ml) observed with a regimen of 1 mg/kg of body weight q24 h . A combination regimen of fluconazole (400 mg q24 h) and amphotericin B (1 mg/kg q24 h) administered simultaneously and as a staggered regimen (amphotericin B bolus given 8 h after fluconazole bolus) was also simulated in the model to characterize possible antagonism between these agents . Fluconazole alone and amphotericin B alone demonstrated fungistatic (< 99.9% reduction in numbers of CFU per milliliter from the starting inoculum) and fungicidal (> 99.9% reduction) activity, respectively . When fluconazole and amphotericin B were administered simultaneously, fungicidal activity similar to that observed with amphotericin B alone was observed . Staggered administration of fluconazole and amphotericin B, however, resulted in a substantial reduction of the fungicidal activity of amphotericin B, producing fungistatic activity similar to that observed with noncombination fluconazole regimens . These results demonstrate the usefulness of this model for comparing the in vitro pharmacodynamic characteristics of different antifungal regimens and support the theory of azole-polyene antagonism . The effects of this antagonism on the in vivo activity and clinical usefulness of combination antifungal therapy, however, remain to be determined.

Arch Biochem Biophys, 1998 May 1, 353(1), 85 - 92
Role of Mg2+ in nucleoside diphosphate kinase autophosphorylation; Biondi RM et al.; Nucleoside diphosphate (NDP) kinase is a ubiquitous enzyme that has been described to have regulatory functions . In addition to its classical enzymatic activity, NDP kinases have been characterized as inhibitors of metastasis, as a factor stimulating gene transcription, and as a protein kinase . In this report we show some characteristics of the autophosphorylation of homogeneous NDP kinase and make a comparison with that of other proteins in crude extracts . By using labeled substrates and fluorescence quenching analysis, we prove that Mg2+ is indeed necessary for the two steps of the ping-pong reaction to take place and present evidence that NTPs or NDPs, when uncomplexed to divalent cations, may not bind the active site in a comparable way to NTP . Mg2+ and NDP . Mg2+ . However, even extremely small concentrations of Mg2+ suffice for maximal autophosphorylation which is obtained with Mg2+ in the nanomolar range and 100 microM ATP using homogeneous enzyme . Moreover, lower autophosphorylation levels were observed with increasing concentrations of Mg2+ . The autophosphorylation equilibrium varied from 0.19 to 1.6 upon the inclusion of 10 mM EDTA to produce low Mg2+ concentrations . Under optimal conditions (low Mg2+ concentrations and short incubation times) NDP kinase was the only protein phosphorylated in crude extracts from Candida albicans, indicating that the autophosphorylation properties of the enzyme are very singular .

Arerugi, 1998 Apr, 47(4), 449 - 56
{Measurement of Candida-specific lymphocyte proliferation by flow cytometry in children with atopic dermatitis}; Kimura M et al.; To study a role of Candida albicans in the development of atopic dermatitis (AD) from the viewpoint of cellular responses, we measured Candida-specific lymphocyte proliferation by flow cytometry in children with AD . There was no apparent age-dependent change in the level of Candida-SIF (stimulation index measured by flow cytometry) in either AD or non-atopic control subjects . The level of Candida-SIF was significantly higher in AD patients than in non-atopic controls (178.0 +/- 89.3 vs 137.9 +/- 37.6, p < 0.02), and the incidence of subjects with the elevated Candida-SIF level (> or = 200) was significantly higher in AD patients than in non-atopic controls (27.9% (17/61) vs 2.6% (1/38), p < 0.005) . There was no correlation between the levels of Candida-SIF and Candida-specific IgE antibody . These results suggest that Candida albicans contributes to the development of AD in some patients not only by Type I, but also by Type IV hypersensitivity reactions.

J Biol Chem, 1998 Jun 5, 273(23), 14392 - 7
The eukaryotic UDP-N-acetylglucosamine pyrophosphorylases . Gene cloning, protein expression, and catalytic mechanism; Mio T et al.; A search of the yeast data base for a protein homologous to Escherichia coli UDP-N-acetylglucosamine pyrophosphorylase yielded UAP1 (UDP-N-acetylglucosamine pyrophosphorylase), the Saccharomyces cerevisiae gene for UDP-N-acetylglucosamine pyrophosphorylase . The Candida albicans and human homologs were also cloned by screening a C . albicans genomic library and a human testis cDNA library, respectively . Sequence analysis revealed that the human UAP1 cDNA was identical to previously reported AGX1 . A null mutation of the S . cerevisiae UAP1 (ScUAP1) gene was lethal, and when expressed under the control of ScUAP1 promoter, both C . albicans and Homo sapiens UAP1 (CaUAP1 and HsUAP1) rescued the ScUAP1-deficient S . cerevisiae cells . All the recombinant ScUap1p, CaUap1p, and HsUap1p possessed UDP-N-acetylglucosamine pyrophosphorylase activities in vitro . The yeast Uap1p utilized N-acetylglucosamine-1-phosphate as the substrate, and together with Agm1p, it produced UDP-N-acetylglucosamine from N-acetylglucosamine-6-phosphate . These results demonstrate that the UAP1 genes indeed specify eukaryotic UDP-GlcNAc pyrophosphorylase and that phosphomutase reaction precedes uridyltransfer . Sequence comparison with other UDP-sugar pyrophosphorylases revealed that amino acid residues, Gly112, Gly114, Thr115, Arg116, Pro122, and Lys123 of ScUap1p are highly conserved in UDP-sugar pyrophosphorylases reported to date . Among these amino acids, alanine substitution for Gly112, Arg116, or Lys123 severely diminished the activity, suggesting that Gly112, Arg116, or Lys123 are possible catalytic residues of the enzyme.

J Leukoc Biol, 1998 Jun, 63(6), 723 - 31
Burn-associated Candida albicans infection caused by CD30+ type 2 T cells; Kobayashi M et al.; Burn injury is associated with the greatly increased susceptibility of thermally injured patients to infection from a variety of pathogens . In this study we investigated the role of burn-associated type 2 T cells on the increased susceptibility of burned patients to Candida albicans infection using SCID mice and peripheral blood lymphocytes (PBL) from thermally injured patients . When SCID mice that were inoculated with PBL from healthy donors were resistant to C . albicans infection, the SCID mice that were inoculated with PBL from burned patients did not show any resistance to the infection . All SCID mice exposed to the pathogen, however, survived after inoculation with patient PBL that were previously depleted of CD30+ cells . The predominance of type 2 T cell responses was demonstrated in PBLs of thermally injured patients . As burn-associated type 2 T cells, CD3+ CD8+ CD30+ IL-4 and IL-10-producing cells were demonstrated in burned patient PBL . These results suggest that burn-associated CD30+ type 2 T cells may play a role on the increased susceptibility of burned patients to C . albicans infection.

J Clin Microbiol, 1998 Jun, 36(6), 1578 - 83
Comparison of three methods for testing azole susceptibilities of Candida albicans strains isolated sequentially from oral cavities of AIDS patients; Tortorano AM et al.; Three susceptibility testing procedures were compared to determine fluconazole, itraconazole, and ketoconazole MICs against 47 Candida albicans strains isolated sequentially from the oral cavities of five AIDS patients undergoing azole therapy . They included the broth microdilution method (BM), performed according to the National Committee for Clinical Laboratory Standards' tentative standard, the agar dilution method (AD), and the Etest; the latter two tests were performed both in Casitone agar (AD-Cas and Etest-Cas) and in RPMI (AD-RPMI and Etest-RPMI) . Twenty-four- and 48-h MICs obtained by AD and Etest were compared with 48-h MICs obtained by BM . The MICs of all the azoles determined by BM were usually lower than those obtained by the other methods, mainly due to different reading criteria . In order to assess the most appropriate way of evaluating the agreement of MICs obtained by different methods with those produced by the proposed reference method (BM), we used the mean differences calculated according to Bland and Altman's method . Comparison of fluconazole MICs obtained by BM and AD-Cas yielded a mean difference of 3, and the percentages of agreement within +/-2 dilutions were 98 and 100% at 24 and 48 h, respectively . For ketoconazole and itraconazole MICs, lower mean differences were noted, and agreement ranged from 96 to 100% . Agreement between the AD-RPMI and BM results was poor for all azoles, and an increase in MICs was always observed between the 1st- and 2nd-day readings . Similarly, Etest-Cas gave better agreement with BM than did Etest-RPMI for all the azoles . BM, AD-Cas, and Etest-Cas each demonstrated a progressive increase in fluconazole MICs against strains isolated sequentially from a given patient, in accordance with the decreased clinical response to fluconazole.

J Clin Microbiol, 1998 Jun, 36(6), 1518 - 29
Hospital specificity, region specificity, and fluconazole resistance of Candida albicans bloodstream isolates; Pfaller MA et al.; In a survey of bloodstream infection (BSI) isolates across the continental United States, 162 Candida albicans isolates were fingerprinted with the species-specific probe Ca3 and the patterns were analyzed for relatedness with a computer-assisted system . The results demonstrate that particular BSI strains are more highly concentrated in particular geographic locales and that established BSI strains are endemic in some, but not all, hospitals in the study and undergo microevolution in hospital settings . The results, however, indicate no close genetic relationship among fluconazole-resistant BSI isolates in the collection, either from the same geographic locale or the same hospital . This study represents the first of three fingerprinting studies designed to analyze the origin, genetic relatedness, and drug resistance of Candida isolates responsible for BSI.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1998 May, 85(5), 558 - 64
Fluconazole-resistant Candida species in the oral flora of fluconazole-exposed HIV-positive patients; Hunter KD et al.; The purpose of this study was to examine the effect of preceding fluconazole treatment on the oral mycologic flora and on the sensitivity of oral Candida albicans isolates to fluconazole . Saline oral rinses were collected from 89 HIV-positive patients, of whom 48 had been exposed to fluconazole and 41 were fluconazole-naive . The rinses were cultured on Sabouraud's and Pagano Levin agars, and yeasts were identified by standard methods . Fluconazole sensitivity of C . albicans isolates was measured by disk diffusion assay . C . albicans was isolated from 69% of patients who had received fluconazole and from 93% of the patients who were fluconazole-naive (p < 0.05) . Nine other species of yeasts were also isolated, most commonly C . glabrata . Five patients previously exposed to fluconazole harbored fluconazole-resistant C . albicans, whereas no resistance was detected among the patients who were fluconazole-naive (p < 0.01) . Sixteen of the patients who were fluconazole-exposed carried yeasts other than C . albicans, compared with only five patients in the fluconazole-naive group (p < 0.01) . All of the fluconazole-resistant strains were isolated from patients with low CD4 counts (less than 100 cells/ml) and after lengthy fluconazole exposures . Nevertheless, patients in Charlotte, N.C., who had a greater mean fluconazole exposure time (10.25 +/- 1.41 months) than patients in Glasgow, UK, (0.65 +/- 0.18 months; p < 0.005), did not develop significantly more in vitro resistance or species diversity . This study indicates that long-term fluconazole treatment can have significant effects on the yeast flora of the mouth, particularly in a patient with a CD4 count of less than 100 cells/ml.

Proc Natl Acad Sci U S A, 1998 Apr 28, 95(9), 5150 - 5
Monosomy of a specific chromosome determines L-sorbose utilization: a novel regulatory mechanism in Candida albicans; Janbon G et al.; We report the identification of the gene, SOU1, required for L-sorbose assimilation in Candida albicans . The level of the expression of SOU1 is determined by the copy number of chromosome III (also denoted chromosome 5), such that monosomic strains assimilate L-sorbose, whereas disomic strains do not, in spite of the fact that SOU1 is not on this chromosome . We suggest that C . albicans contains a resource of potentially beneficial genes that are activated by changes in chromosome number, and that this elaborate mechanism regulates the utilization of food supplies and possibly other important functions, thus representing a novel general means for regulating gene expression in microbes.

J Dent, 1998 May, 26(4), 299 - 304
Denture plaque--past and recent concerns; Nikawa H et al.; OBJECTIVE: This paper critically reviews the history of denture plaque and identifies some concerns with the presence of Candida in the mouth . DATA SOURCES: This review covers literature sources related to Candida albicans and its relationship to denture plaque . STUDY SELECTION: The articles selected for this review are from referred journals and describe C . albicans and its relationship to oral, gastrointestinal and pleuropulmonary infections . The relationship to caries, root caries and periodontal disease is also covered . CONCLUSIONS: Denture plaque containing Candida could cause not only oral candidiasis, like oral thrush or denture-induced stomatitis, but also caries, root caries and periodontitis of abutment teeth . However, there is only limited experimental evidence or information available on the cariogenicity of Candida . The continuous swallowing or aspiration of micro-organisms from denture plaque exposes patients, particularly the immunocompromised host or medicated elderly, to the risks of unexpected infections . The term, 'denture plaque' has been used throughout the review . However, the term 'plaque on denture' should be used because the microbial flora and its pathogenicity of denture plaque resembles those of plaque formed on the tooth surface, so called dental plaque . In addition, the term 'denture related stomatitis' would be preferable to 'denture induced stomatitis', since the inflammation of (palatal) mucosa is not induced by the denture, but by wearing the denture or by plaque on the denture.

Boll Chim Farm, 1998 Mar, 137(3), 93 - 6
Studies on arylmethyltriazinone derivatives-Part II . Synthesis and antifungal properties of 2-aminomethyl-4-(arylidene)amino-6-arylmethyl- 3-mercapto-1,2,4-triazin-5(4H)-ones; Holla BS et al.; The preparation of twenty new 2-aminomethyl-4-(arylidene)amino- 6-arylmethyl-3-mercapto-1,2,4-triazin-5(4H)-ones(4) is described . The structural elucidation of all the compounds is carried out on the basis of analytical and spectral data . The newly synthesized compounds are evaluated for their antifungal activity against Candida albicans and Paecileomyces variotti.

Phytochemistry, 1998 Apr, 47(7), 1223 - 6
Secoiridoid glycosides and an antifungal anthranilate derivative from Gentiana tibetica; Tan RX et al.; Repetitive chromatography of the methanol extract of the roots of Gentiana tibetica afforded two new secoiridoid glycosides and a novel antifungal anthranilic acid derivative, together with beta-sitosterol, daucosterol, oleanolic acid, loganic acid, gentiopicroside, sweroside, 2'-(2,3-dihydroxybenzoyl)sweroside, trifloroside, rindoside and macrophylloside A . The structures of the new products were determined mainly by spectroscopic methods as 8-hydroxy-10-hydrosweroside, isomacrophylloside and ethyl N-docosanoylanthranilate . Ethyl N-docosanoylanthranilate inhibited the growth of the human pathogenic fungi Candida albicans and Aspergillus flavus . The taxonomic significance of the constituent is discussed briefly.

Urology, 1998 May, 51(5), 846 - 8
Hepatic subcapsular extension of pelvic lymphocele after radical retropubic prostatectomy; Bauer JJ et al.; Lymphoceles are a rare symptomatic complication after radical prostatectomy occurring in less than 5% of cases . We present a case of a symptomatic lymphocele that occurred after a radical retropubic prostatectomy and obturator lymphadenectomy . The lymphocele dissected along the retroperitoneum and extended into the hepatic subcapsular space and became secondarily infected with Candida albicans.

J Infect Dis, 1998 Jun, 177(6), 1660 - 3
Interactions of itraconazole with amphotericin B in the treatment of murine invasive candidiasis; Sugar AM et al.; The interactions of amphotericin B and itraconazole were studied in murine invasive candidiasis . Candida albicans-infected mice were treated for 10 consecutive days, 24 h after infection . Survival was monitored over 30 days and kidney cultures were done . Mice treated with amphotericin B (0.2 mg/kg/day intraperitoneally) or itraconazole (100 mg/kg/day by oral gavage in two divided doses/ day) had a 30-day survival of 20% or 40% . Concomitant administration of both drugs resulted in 100% mortality; 90% of mice treated with amphotericin B (1 mg/kg/day) survived . With the combination, 100% were dead by day 28 (P < or = .001 vs . amphotericin B) . With sequential therapy (i.e., 5 days with one drug and then 5 days with the other), survival was inferior to that with amphotericin B alone but similar to that with itraconazole alone . Kidney culture results confirmed the antagonism of the combination compared with amphotericin B alone . In treatment of murine invasive candidiasis, the concomitant or sequential use of amphotericin B and itraconazole results in a negative interaction.

J Inorg Biochem, 1998 Feb 1, 69(1-2), 15 - 23
Antifungal nickel(II) complexes derived from amino sugars against pathogenic yeast, Candida albicans; Yano S et al.; Nickel(II) complexes containing N-glycosides derived from D-glucosamine (D-GlcN) and ethylenediamine (en) and trimethylenediamine (tn), {Ni(D-GlcN-en)2}Cl2.H2O (1) (D-GlcN-en = 1- inverted question mark(2-aminoethyl)amino inverted question mark-2-amino-1,2-dideoxy-D-glucose) and {Ni(D-GlcN-tn)2}Cl2.4H2O (2) (D-GlcN-tn = 1- inverted question mark(3-aminopropyl)amino inverted question mark-2-amino-1,2-dideoxy-D-glucose), are fairly stable in water at room temperature and showed effective antifungal activity against pathogenic yeast, Candida albicans, with the MIC (minimal concentration of inhibition) values of the complexes being 0.25 mM . The results obtained enzyme assays by using preparations of C . albicans chitinase fraction suggested that the sugar complexes 1 and 2 played a role of novel chitinase (chitin-degradation enzyme) inhibitor, where the modes of inhibition were competitive (Ki = 1.3 mM for 1, Ki = 1.8 mM for 2) . The newly prepared nickel(II) complex 2 was characterized by elemental analysis, magnetic susceptibility, electronic absorption and circular dichroism spectroscopies, and an X-ray crystallographic analysis.

Z Arztl Fortbild Qualitatssich, 1998 Apr, 92(3), 175 - 9
{Vulvovaginal mycoses}; Mendling W; Due to its pathogenety Candida albicans is the most frequent yeast in cases of vaginal candidosis, probably mostly caused by local immunological weakness . In 5-30% one can expect a vaginal yeast colonisation depending on age, estrogen influence, pregnancy and dispositions by illness . Prepartal vaginal yeast colonisation should be treated to protect the newborn . The only typical symptom of acute vaginal candidosis is itching . Beside history and clinical symptoms, examination of vaginal secretion by phase contrast microscopy and the yeast culture are cornerstones of the diagnosis . Antimycotic resistance should be investigated only by specialists . Acute Candida albicans vaginitis should be treated locally by one or three day therapy . Candida glabrata vaginitis can be treated with high doses of oral fluconazole.

Z Arztl Fortbild Qualitatssich, 1998 Apr, 92(3), 163 - 8
{Candida infections in infancy and early childhood}; Schwarze R et al.; Candida infections in infancy can manifest themselves as skin, mucosal or systemic candidiasis . Eighty to nintey percent of all candida infections in this age group are caused by Candida albicans . Whereas in neonates, infections mostly occur sub partu, in older children predisposing underlying diseases get an increasing etiological importance . The diagnosis is based on microscopic and cultural detection of yeast as well as on the course of the titers of Candida antigen and antibodies . For topical antifungal treatment of skin and mucosa infections, different preparations of the polyenes nystatin and amphotericin B have been proven to be most effective . In systemic candidiasis the combination of amphotericin B and 5-flucytosin is the treatment of choice . In view of the potential severe side effects of this combination therapy, fluconazol as a sole treatment represents an effective alternative . Prophylaxis against Candida infections comprises sticking to hygienic regimes, mycological surveillance of risk groups and oral application of antimycotics.

Z Arztl Fortbild Qualitatssich, 1998 Apr, 92(3), 157 - 62
{Fungi in the oro-intestinal tract and their scientifically founded status}; Knoke M; The orointestinal tract is a reservoir for facultatively pathogenic fungi, especially Candida albicans . In all of its sections in immunocompromised hosts, the occurrence of a mucosal mycosis is possible which may be the starting point of an infection of internal organs . The mouth and esophagus are the most often affected locations . A synopsis of clinical (including endoscopic) findings, mycological cultivation and mycoserology is important in diagnostics . There is no connection between the incidence of Candida in the orointestinal tract and multiple local symptoms like fatigue, headache, heartburn and others called "candidiasis hypersensitivity syndrome" or "mycophobia".

Yeast, 1998 Apr 30, 14(6), 565 - 71
An improved protocol for the preparation of yeast cells for transformation by electroporation; Thompson JR et al.; Pretreatment of yeast cells with lithium acetate (LiAc) and dithiothreitol (DTT) enhances the frequency of transformation by electroporation . The method shows improvements of 6-67-fold in wild-type strains derived from commonly used Saccharomyces cerevisiae genetic backgrounds . In addition, 15-300-fold improvement in transformation frequency was achieved with several mutant strains of S . cerevisiae that transformed poorly by conventional procedures . Both DTT and lithium acetate were necessary for maximal transformation frequencies . Pretreatment with lithium and DTT also resulted in an approximately 3-5-fold increase in the electroporation transformation frequency of the pathogenic fungus Candida albicans.

Yeast, 1998 Apr 30, 14(6), 535 - 50
Deletion of transmembrane domain 12 of CDR1, a multidrug transporter from Candida albicans, leads to altered drug specificity: expression of a yeast multidrug transporter in baculovirus expression system; Krishnamurthy S et al.; Cdr1p, an ATP-binding cassette transporter from the pathogenic yeast Candida albicans, confers resistance to several unrelated drugs including anti-Candida drugs (Prasad et al., 1995b) . We demonstrate that the deletion of 237 bp (79 aa) from the 3' end of CDR1 (which encompasses the transmembrane domain (TM) 12 of the putative transporter) did not result in the total loss of its ability to efflux cytotoxic agents . While the expression of deltaCDR1 in yeast resulted in impaired sensitivity to drugs like cycloheximide, anisomycin, sulfomethuron methyl and antifungal nystatin, its ability to confer resistance remained unaltered to drugs such as o-phenanthroline, 4-nitroquinoline-N-oxide, cerulenin, azoles, oligomycin, erythromycin, and benomyl . Similar to human MDR1p . Cdr1p might also have localized drug binding sites in TM 12, but that might not be the case for all the drugs . The TM 12 deletion also did not lead to any significant impairment in NTPase activities . Both ATPase and UTPase activities of complete Cdr1p and deltaCdr1p were not significantly altered, as was the case with respect to their ability to efflux Rh123 and steroid hormone like {3H}-beta-estradiol . To further dissect the functionality of Cdr1p, its truncated version was overexpressed in a baculovirus-insect cell expression system . The synthesis of deltaCdr1p in Sf9 cells was temporally regulated as a function of the baculovirus polyhedrin gene promoter . The Sf9 derived deltaCdr1p was approximately 130 kDa, which was lower than the expected size, probably due to the differences in glycosylation . This, however, did not affect the functionality of deltaCdr1p . The deletion of TM 12 did not affect the targeting of the protein and deltaCdr1p was exclusively localized in plasma membrane of Sf9 cells as detected by immunofluorescence . The expression of deltaCdr1p in the baculovirus-insect expression system generated a high drug-stimulated plasma membrane-bound ATPase activity which was not demonstrable when deltaCdr1p was expressed in yeast.

Yeast, 1998 Apr 30, 14(6), 517 - 26
Induced expression of the Candida albicans multidrug resistance gene CDR1 in response to fluconazole and other antifungals; Hernaez ML et al.; The Candida albicans CDR1 gene encodes a member of the ABC-type family of multidrug transporters which has been shown to be involved in azole resistance . Using an in-frame gene fusion between the CDR1 open reading frame and the green fluorescent protein allele yEGFP3, an optimized derivative for its use in C . albicans, we show here how the CDR1-yEGFP3 gene expression is induced in response to azoles as well as to other structurally unrelated drugs like cycloheximide . Moderate increases were observed for calcofluor, canavanine, 5'-fluorcytosine, cilofungin and caffeine, while no induction was found for the antifungals benomyl and amphotericin B or hydrogen peroxide at subinhibitory concentrations . The use of confocal microscopy enabled us to localize the Cdr1p fusion protein at the cell periphery, thus suggesting a cytoplasmic membrane localization . These results suggest deregulation of CDR1 gene as a putative mechanism for the generation of azole resistance in this clinically important pathogenic fungus.

Clin Diagn Lab Immunol, 1998 May, 5(3), 369 - 74
Detection of antibodies to Candida albicans germ tubes during experimental infections by different Candida species; Bikandi J et al.; Identification and characterization of Candida albicans germ tube-specific antigens may be of relevance for the serodiagnosis of invasive candidiasis since they could be the basis for the development of new diagnostic tests . In this study, we have identified two antigens of 180 and >200 kDa in the cell wall of C . albicans germ tubes which are responsible for the induction of antibodies to C . albicans germ tubes . Antigens of similar molecular masses have been demonstrated in the cell walls of the Candida species C . stellatoidea, C . parapsilosis, C . guilliermondii, C . tropicalis, and C . krusei, but not C . glabrata . The kinetics of the antibody responses to C . albicans germ tubes were studied in rabbits infected with different Candida species . Although these antibodies were detected in rabbits infected with all Candida species except C . glabrata, the kinetics of the antibody responses to C . albicans germ tubes induced by the Candida species studied were different . Both the highest titer and the earliest response of antibodies to C . albicans germ tubes were observed in rabbits infected with either of the two serotypes of C . albicans used . However, the time needed to elicit the antibodies to C . albicans germ tubes can be reduced as the result of an anamnestic antibody response . The results presented in this study show that a test designed to detect antibodies against C . albicans germ tube antigens may be suitable for the diagnosis of infections caused by most of the medically important Candida species.

Clin Diagn Lab Immunol, 1998 May, 5(3), 335 - 40
Cytokine gene expression in normal human lymphocytes in response to stimulation; Fan J et al.; Sequential gene expression of two type 1 cytokines (interleukin 2 {IL-2} and gamma interferon), one type 2 cytokine (IL-10), two monokines (IL-6 and tumor necrosis factor alpha), and one cytokine receptor (IL-2 receptor {IL-2R}) in normal human peripheral blood mononuclear cells (PBMC) following in vitro stimulation was investigated by reverse transcription-PCR methods . Two stimuli were utilized: phytohemagglutinin (PHA), which acts on the CD2 molecule and T-cell receptors, and anti-CD3 monoclonal antibody, which acts on the CD3 molecule and on T-cell receptors . Increased expression of all studied genes occurred between 1 and 4 hours after stimulation, except for that of the gene encoding IL-10, which was delayed . Expression of all but one of the genes was transient, with a maximal mRNA accumulation at about 8 h on average . IL-2R mRNA expression was an exception, showing a prolonged increase (72 h) . The general profiles of expression of the five cytokine genes were similar but not identical, suggesting some shared regulatory mechanisms . When responses to four additional stimuli (pokeweed mitogen, Candida albicans, and IL-2 at high and low doses) were compared, similar profiles of cytokine gene expression were found . Thus, the various stimuli caused induction of all cytokines with quantitative, not qualitative, differences . Altogether, the present data are useful for defining the kinetics of gene expression for key cytokines in response to standard immune-cell stimuli.

Proc Natl Acad Sci U S A, 1998 May 26, 95(11), 5857 - 64
SMART, a simple modular architecture research tool: identification of signaling domains; Schultz J et al.; Accurate multiple alignments of 86 domains that occur in signaling proteins have been constructed and used to provide a Web-based tool (SMART: simple modular architecture research tool) that allows rapid identification and annotation of signaling domain sequences . The majority of signaling proteins are multidomain in character with a considerable variety of domain combinations known . Comparison with established databases showed that 25% of our domain set could not be deduced from SwissProt and 41% could not be annotated by Pfam . SMART is able to determine the modular architectures of single sequences or genomes; application to the entire yeast genome revealed that at least 6.7% of its genes contain one or more signaling domains, approximately 350 greater than previously annotated . The process of constructing SMART predicted (i) novel domain homologues in unexpected locations such as band 4.1-homologous domains in focal adhesion kinases; (ii) previously unknown domain families, including a citron-homology domain; (iii) putative functions of domain families after identification of additional family members, for example, a ubiquitin-binding role for ubiquitin-associated domains (UBA); (iv) cellular roles for proteins, such predicted DEATH domains in netrin receptors further implicating these molecules in axonal guidance; (v) signaling domains in known disease genes such as SPRY domains in both marenostrin/pyrin and Midline 1; (vi) domains in unexpected phylogenetic contexts such as diacylglycerol kinase homologues in yeast and bacteria; and (vii) likely protein misclassifications exemplified by a predicted pleckstrin homology domain in a Candida albicans protein, previously described as an integrin.

J Med Chem, 1998 May 7, 41(10), 1631 - 40
Syntheses and biological evaluation of indolocarbazoles, analogues of rebeccamycin, modified at the imide heterocycle; Moreau P et al.; A series of 10 indolocarbazole derivatives, analogues to the antitumor antibiotic rebeccamycin, bearing modifications at the imide heterocycle were synthesized . They bear an N-methyl imide, N-methyl amide, or anhydride function instead of the original imide . Their inhibitory potencies toward topoisomerase I were examined using a DNA relaxation assay and by analyzing the drug-induced cleavage of 32P-labeled DNA . Protein kinase C (PKC) inhibition and interaction with DNA were also studied together with the in vitro antiproliferative activities against B16 melanoma and P388 leukemia cells . The antimicrobial activities against two Gram-positive bacteria (Bacillus cereus and Streptomyces chartreusis), a Gram-negative bacterium (Escherichia coli), and a yeast (Candida albicans) were tested as well as their antiviral activities toward HIV-1 . The efficiency of the anhydride compounds was compared to that of the parent compound rebeccamycin and its dechlorinated analogue . All the compounds studied were inactive against PKC . The structural requirements for PKC and topoisomerase I inhibition are markedly different . In sharp contrast with the structure-PKC inhibition relationships, we found that an anhydride function does not affect topoisomerase I inhibition, whereas a methyl group on the indole nitrogen prevents the poisoning of topoisomerase I . The compounds exhibiting a marked toxicity to P388 leukemia cells had little or no effect on the growth of P388CPT5 cells which are resistant to the topoisomerase I inhibitor camptothecin . This study reinforces the conclusion that the DNA-topoisomerase I cleavable complex is the primary cellular target of the indolocarbazoles and significantly contributes to their cytotoxicity and possibly to their weak but noticeable anti-HIV-1 activities . The structure-activity relationships are also discussed.

Infect Immun, 1998 Jun, 66(6), 3003 - 5
Secreted aspartyl proteinases and interactions of Candida albicans with human endothelial cells; Ibrahim AS et al.; The endothelial cell interactions of homozygous null mutants of Candida albicans that were deficient in secreted aspartyl proteinase 1 (Sap1), Sap2, or Sap3 were investigated . Only Sap2 was found to contribute to the ability of C . albicans to damage endothelial cells and stimulate them to express E-selectin . None of the Saps studied appears to play a role in C . albicans adherence to endothelial cells.

Infect Immun, 1998 Jun, 66(6), 2750 - 4
Augmentation of human macrophage candidacidal capacity by recombinant human myeloperoxidase and granulocyte-macrophage colony-stimulating factor; Marodi L et al.; Phagocyte myeloperoxidase (MPO) is believed to be particularly important in defense against candida infection . We reported earlier that monocytes, rich in MPO, killed Candida albicans at a significantly higher rate and extent than did monocyte-derived macrophages, known to lack MPO, and that C . albicans is less resistant to MPO-dependent oxidants than less pathogenic Candida species . We hypothesized, therefore, that the capacity of macrophages to kill C . albicans might be improved in the presence of MPO . In this study, we evaluated the ability of recombinant human MPO (rhMPO) to augment the killing of C . albicans by resident macrophages and macrophages activated by recombinant human granulocyte-macrophage colony-stimulating factor . Addition of rhMPO (concentration range, 0.8 to 6.4 U/ml) to suspensions of resident and activated macrophages and opsonized C . albicans resulted in concentration-dependent and significant increases in candida killing . This enhancement was particularly pronounced with activated macrophages, whether C . albicans was opsonized or unopsonized and ingested through the macrophage mannose receptor . rhMPO did not affect the killing of C . albicans by monocytes, nor did it affect phagocytosis of opsonized or unopsonized C . albicans . These results indicate that exogenous rhMPO can augment the candidacidal capacity of both resident and activated macrophages, with a more profound effect on activated cells . We suggest that rhMPO may be effective in the treatment of invasive candidiasis.

Infect Immun, 1998 Jun, 66(6), 2713 - 21
Roles of the Candida albicans mitogen-activated protein kinase homolog, Cek1p, in hyphal development and systemic candidiasis; Csank C et al.; Extracellular signal-regulated protein kinase (ERK, or mitogen-activated protein kinase {MAPK}) regulatory cascades in fungi turn on transcription factors that control developmental processes, stress responses, and cell wall integrity . CEK1 encodes a Candida albicans MAPK homolog (Cek1p), isolated by its ability to interfere with the Saccharomyces cerevisiae MAPK mating pathway . C . albicans cells with a deletion of the CEK1 gene are defective in shifting from a unicellular budding colonial growth mode to an agar-invasive hyphal growth mode when nutrients become limiting on solid medium with mannitol as a carbon source or on glucose when nitrogen is severely limited . The same phenotype is seen in C . albicans mutants in which the homologs (CST20, HST7, and CPH1) of the S . cerevisiae STE20, STE7, and STE12 genes are disrupted . In S . cerevisiae, the products of these genes function as part of a MAPK cascade required for mating and invasiveness of haploid cells and for pseudohyphal development of diploid cells . Epistasis studies revealed that the C . albicans CST20, HST7, CEK1, and CPH1 gene products lie in an equivalent, canonical, MAPK cascade . While Cek1p acts as part of the MAPK cascade involved in starvation-specific hyphal development, it may also play independent roles in C . albicans . In contrast to disruptions of the HST7 and CPH1 genes, disruption of the CEK1 gene adversely affects the growth of serum-induced mycelial colonies and attenuates virulence in a mouse model for systemic candidiasis.

Infect Immun, 1998 Jun, 66(6), 2640 - 7
Interleukin-15 activates proinflammatory and antimicrobial functions in polymorphonuclear cells; Musso T et al.; Interleukin-15 (IL-15) is a recently discovered cytokine produced by a wide range of different cell types including fibroblasts, keratinocytes, endothelial cells, and macrophages in response to lipopolysaccharide or microbial infection . This suggests that IL-15 may play a crucial role in the activation of phagocytic cells against pathogens . We studied polymorphonuclear leukocyte (PMN) activation by IL-15, evaluated as enhancement of PMN anti-Candida activity as well as IL-8 production, following stimulation with the cytokine . The PMN response to IL-15 depends on binding to the IL-15 receptor . Our experiments show that binding of a biotinylated human IL-15-immunoglobulin G2b IgG2b fusion protein was competed by the addition of human recombinant IL-15 (rIL-15) or of human rIL-2, suggesting that IL-15 binding to PMN might involve the IL-2Rbeta and IL-2Rgamma chains, which have been shown to be constitutively expressed by PMN . In addition, we show by reverse transcription-PCR and by flow cytometry with a specific anti-IL-15Ralpha chain monoclonal antibody that PMN express the IL-15Ralpha chain at the mRNA and protein levels . Incubation with IL-15 activated PMN to secrete the chemotactic factor IL-8, and the amount secreted was increased by costimulation with heat-inactivated Candida albicans . In addition, IL-15 primed the metabolic burst of PMN in response to formyl-methionyl-leucyl-phenylalanine but was not sufficient to trigger the respiratory burst or to increase the production of superoxide in PMN exposed to C . albicans . IL-15 also increased the ability of PMN to phagocytose heat-killed C . albicans organisms in a dose-dependent manner, without opsonization by antibodies or complement-derived products . In the same concentration range, IL-15 was as effective as gamma interferon (IFN-gamma) and IL-2 in increasing the C . albicans growth-inhibitory activity of PMN . Taken together, these results suggest that IL-15 is a potent stimulant of both proinflammatory and antifungal activities of PMN, activating several antimicrobial functions of PMN involved in the cellular response against C . albicans.

Infect Immun, 1998 Jun, 66(6), 2486 - 93
Activity of protegrins against yeast-phase Candida albicans; Cho Y et al.; We used a two-stage radial diffusion assay to perform a structure-activity study of the antifungal effects of protegrin-1 (PG-1) on yeast-phase Candida albicans . While doing so, we computed MICs from the radial diffusion assay data by three methods and compared the respective values with results from colony count and broth microdilution assays . This allowed us to identify several technical modifications that improved the sensitivity and accuracy of radial diffusion assays . We found that both PG-1 and enantiomeric PG-1 (composed exclusively of D-amino acids) were potently fungicidal for yeast-phase C . albicans . The protegrins PG-2, -3, and -5, but not PG-4, were as effective as PG-1 . At least one intramolecular disulfide bond was required to retain optimal candidacidal activity at physiological NaCl concentrations . Truncated variants of PG-1 that lacked its first four residues showed decreased candidacidal activity, although their activity against bacteria was substantially intact . Altering the beta-turn region (residues 9 to 12) of PG-1 or its variants further decreased candidacidal activity . These studies suggest that only 12 residues are needed to endow protegrin molecules with strong antibacterial activity and that at least 4 additional residues are needed to add potent antifungal properties . Thus, the 16-residue protegrin PG-2 likely represents the minimal structure needed for broad-spectrum antimicrobial activity encompassing bacteria and fungi.

Radiats Biol Radioecol, 1997 Jul-Aug, 37(4), 649 - 56
{Fungal infection of human organs by resistant melanin-synthesizing species is one of the pathogenic factors and one of the real consequences of the accident at Chernobyl power plant}; Pulatova MK et al.; Free radical melanin centers have been detected in the cell concentrate of bronchoalveolar lavage (BAL) of liquidator of Chernobyl NPP accident . To identify the nature of these centers the EPR technique and the fluorescent technique were used to study BAL of liquidators with lung chronic pathology, their blood, blood components as well as model melanin- and lipofuscin-containing systems: synthetic DOPA-melanin, human melanosome, human lipofuscin, human melanolipofuscin . I Besides that we have investigated the samples of fungi, extracted from lung phlegm of liquidators (Aspergillus niger, Aspergillus fumigatus, Aspergillus flavus, Penicillium Sp., Candida albicans) as well as the melanin, extracted from fungal conidium . It has been shown that the melanin centers found in BAL cells of liquidators is the melanin of melanin-synthesizing mutant fungi Aspergillus fumigatus and Aspergillus flavus . The prolonged gamma-irradiation at low dose rate and the effects of inhaled radioactive particles cause the adaptive mutation of micromycetes producing the chemo- and radioresistant population . We think that the radioactive dust and pathogenic mutant micromycetes were inhaled in lungs of liquidators during their work at the Chernobyl NPP . Thus, one of valid consequences of Chernobyl accident may be the wide fungous of human organs, in particularly, by Aspergillus mutant . The radiation-induced weakening of immune reactions of liquidators promotes the resistance of this fungus mutant infection.

Curr Genet, 1998 Apr, 33(4), 268 - 75
Molecular analysis of the LYS2 gene of Candida albicans: homology to peptide antibiotic synthetases and the regulation of the alpha-aminoadipate reductase; Suvarna K et al.; The unique alpha-aminoadipate pathway for lysine biosynthesis is present only in fungi and involves eight enzyme steps . alpha-Aminoadipate semialdehyde dehydrogenase, commonly called alpha-aminoadipate reductase (AAR), catalyzes the conversion of alpha-aminoadipic acid to alpha-aminoadipic semialdehyde by a novel mechanism . Two genes, LYS2 and LYS5, encode the heterodimeric enzyme in Saccharomyces cerevisiae . The LYS2 gene of Candida albicans was shown to be contained in the 4.8-kb insert of the plasmid pCaLYS2 . This plasmid complemented lys2 mutants of both S . cerevisiae and C . albicans . The S . cerevisiae and C . albicans Lys2(+) transformants exhibited 138% and 160% of wild-type AAR activity, respectively . The DNA-sequence analysis of the 4.8-kb region in plasmid pCaLYS2 and a PCR product from genomic DNA which overlapped with the 4.8-kb insert revealed a continuous ORF of 4173 nucleotides encoding 1391 amino-acid residues . The C . albicans LYS2 ORF exhibited 63.0% identity at the nucleotide level and 56.2% identity at the amino-acid level to the LYS2 gene of S . cerevisiae . The ORF is preceded by consensus sequences for the TATA-, CAAT- and GCN4-box elements . An S . cerevesiae-type transcription termination signal is seen in the 3' flanking region . The deduced amino-acid sequence revealed a motif for an AMP-binding site and also the highly conserved core sequences common to peptide antibiotic synthetases . The LYS2 mRNA and alpha-aminoadipate reductase activity were repressed to a higher level in YEPD-grown cells than in cells grown in the presence of lysine or minimal medium . Additionally, AAR was shown to be feedback-inhibited by lysine and the lysine analog, thialysine . The results of the present report reveal the molecular characteristics of the LYS2 gene of C . albicans, its homology to peptide antibiotic synthetases, its divergence from the LYS2 gene of S . cerevisiae, and the regulation of AAR in C . albicans.

Diagn Microbiol Infect Dis, 1998 May, 31(1), 327 - 32
National surveillance of nosocomial blood stream infection due to Candida albicans: frequency of occurrence and antifungal susceptibility in the SCOPE Program; Pfaller MA et al.; Surveillance of nosocomial blood stream infections (BSI) in the USA between April 1995 and June 1996 revealed that Candida was the fourth leading cause of nosocomial BSI, accounting for 8% of all infections . Fifty-two percent of 379 episodes of candidemia were due to Candida albicans . In vitro susceptibility studies using the 1997 National Committee for Clinical Laboratory Standards reference method demonstrated that 92% of C . albicans isolates were susceptible to 5-fluorocytosine and 90% were susceptible to fluconazole and itraconazole . Geographic variation in susceptibility of fluconazole and itraconazole was observed . Isolates from the Northwest and Southeast regions were more frequently resistant to fluconazole (13.3-15.5%) and to itraconazole (17.2-20.0%) than those from the Northeast and Southwest regions (2.9-5.5% resistant to fluconazole and itraconazole) . Continued surveillance for infections caused by C . albicans and other species of Candida among hospitalized patients is recommended.

Diagn Microbiol Infect Dis, 1998 May, 31(1), 297 - 300
In vitro activity of voriconazole against Candida species; Kauffman CA et al.; The in vitro activity of voriconazole was compared with that of itraconazole and fluconazole against 181 isolates of Candida albicans, 124 isolates of Candida glabrata, and 20 isolates of Candida krusei obtained from the early 1980s through the mid-1990s . Voriconazole had greater intrinsic activity than fluconazole or itraconazole against all three Candida species . For C . glabrata, C . krusei, and C . albicans, the MIC50 values for voriconazole were 1 microgram/mL, 0.5 microgram/mL, and 0.01 microgram/mL, respectively compared with fluconazole MIC50 values of 8 micrograms/mL, 64 micrograms/mL, and 0.25 microgram/mL, respectively . If isolates from AIDS patients were excluded, MIC values for isolates from the 1990s were no higher than those noted for isolates from the 1980s . Voriconazole, a new triazole antifungal agent, appears to have enhanced activity against these three species of Candida; the clinical relevance of these findings should be studied in treatment trials.

Clin Infect Dis, 1998 May, 26(5), 1134 - 41
Candidemia: a nosocomial complication in adults with late-stage AIDS; Launay O et al.; We retrospectively analyzed 13 episodes of candidemia observed between July 1990 and July 1995 in human immunodeficiency virus (HIV)-infected adults . Candidemia was nosocomially acquired by 11 patients, among whom nine had a central venous catheter (CVC) . Twelve cases were of stage C2/C3 according to the 1993 classification of the Centers for Disease Control and Prevention . The median CD4+ cell count was 10/mm3 (range, 3-400/mm3) . Causative species were Candida albicans in nine episodes and Candida glabrata and Candida krusei in two episodes each . Eleven episodes occurred in 11 patients who had previously received fluconazole (mean total dose, 7.4 g), including the four episodes caused by non-albicans species . Outcome did not differ according to the administered antifungal therapy . CVCs were removed from seven patients (78%) . The overall mortality was 38% . Candidemia is a potentially lethal nosocomial complication during late-stage AIDS and can be due to C . albicans and non-albicans strains.

J Clin Microbiol, 1998 Apr, 36(4), 1157 - 9
Chromogenic tube test for presumptive identification or confirmation of isolates as Candida albicans; Merlino J et al.; This report describes a new, modified, simple, and cost-effective method for the use of CHROMagar Candida (CHROMagar Company, Paris, France) for the presumptive identification of isolates as Candida albicans after preliminary growth . Sixty randomly selected clinical isolates were evaluated, including 38 of C . albicans . With incubation at 37 degrees C for 24 h, the sensitivity and specificity appeared to be excellent and the test performed better than the traditional germ tube test . However, at earlier times, C . tropicalis isolates gave false-positive results.

J Clin Microbiol, 1998 Apr, 36(4), 1035 - 8
Intergenic transcribed spacer PCR ribotyping for differentiation of Saccharomyces species and interspecific hybrids; McCullough MJ et al.; The taxonomy of the genus Saccharomyces has undergone significant changes recently with the use of genotypic rather than phenotypic methods for the identification of strains to the species level . The sequence of rRNA genes has been utilized for the identification of a variety of fungi to the species level . This methodology, applied to species of Saccharomyces, allows unknown Saccharomyces isolates to be assigned to the type strains . It was the aim of the present study to assess whether typing of the intergenic spacer region by using restriction fragment length polymorphisms of PCR products (intergenic transcribed spacer PCR {ITS-PCR} ribotyping) could distinguish among type strains of the 10 accepted species of Saccharomyces and further to assess if this method could distinguish strains that were interspecific hybrids . Cellular DNA, isolated after the lysis of protoplasts, was amplified by PCR using ITS1 and ITS4 primers, purified by liquid chromatography, and digested with restriction endonucleases . Ribotyping patterns using the restriction enzymes MaeI and HaeIII could distinguish all species of Saccharomyces from each other, as well as from Candida glabrata, Candida albicans, and Blastomyces dermatitidis . The only exception to this was the inability to distinguish between Saccharomyces bayanus and S . pastorianus (S . carlsbergensis) . Furthermore, interspecific hybrids resulting from the mating of sibling species of Saccharomyces were shown to share the ITS-PCR ribotyping patterns of both parental species . It should now be possible, by this simple PCR-based technique, to accurately identify these strains to the species level, thereby allowing an increase in our understanding of the characteristics required by these interspecific hybrids for their particular ecological niches.

Ann Ist Super Sanita, 1997, 33(3), 343 - 5
Diabetes and pregnancy: prophylaxis of genital infections; Mazziotti F et al.; Diabetic pregnant women have many potential reasons to have genital infections such as poor metabolic control and impairement of leucocyte function . Relative immune deficiency exists in pregnancy . This study was designed to test the hypotheses that pregnant patients with insulin-dependent diabetes have a higher rate of ante partum genital infections when compared with a pair-matched control population . Two groups of pregnant women consisting of 23 patients with and 23 without diabetes mellitus, underwent colposcopy and cytology between 16th and 24th week of gestation to detect the presence of human papilloma virus (HPV), Gardnerella vaginalis, Candida albicans and aspecific infections . No significant differences were observed between the two groups.

J Am Acad Dermatol, 1998 May, 38(5 Pt 3), S32 - 36
Causative pathogens in onychomycosis and the possibility of treatment resistance: a review; Evans EG; Onychomycosis is caused primarily by dermatophytes, Candida species, and nondermatophytic molds . Dermatophytes, particularly Trichophyton rubrum, are by far the most common pathogens . There is some question as to whether Candida actually breaks down nail material or only invades the proximal nail bed . Similarly, the clinical significance of molds is unknown, because they may be colonizing organisms rather than destructive pathogens . It is, therefore, important to identify the pathogen in the array of organisms that may be isolated in culture . The increasing use of fluconazole in prophylaxis and treatment of systemic yeast infections and infections in patients with AIDS has been associated with the emergence of resistant Candida albicans, as well as previously minority species of Candida, such as C . glabrata and C . krusei . This may be of relevance to the treatment of onychomycosis with azole antifungals.

Antimicrob Agents Chemother, 1998 May, 42(5), 1213 - 6
In vitro antimicrobial activity of MSI-78, a magainin analog; Fuchs PC et al.; MSI-78 is a cationic peptide with broad-spectrum antimicrobial activity and is being developed as a topical agent . We compared the in vitro activity of MSI-78 with those of ofloxacin and other antibiotics against fresh clinical isolates . Based on MIC distribution statistics, strains for which the MSI-78 MIC was < or = 64 micro/ml were assumed to be susceptible for purposes of this report . Of 411 aerobic isolates tested, 91% were susceptible to MSI-78, compared to 91% for ofloxacin and 92% for ciprofloxacin . Only enterococci consistently required > or = 64 microg of MSI-78/ml for inhibition . MSI-78 demonstrated bactericidal activity equivalent to that of ofloxacin . Of 61 anaerobes, 97% were susceptible to MSI-78 . Of 10 isolates of Candida albicans, 3 were inhibited by MSI-78 at 24 h . Further studies of this compound appear to be warranted.

Antimicrob Agents Chemother, 1998 May, 42(5), 1187 - 94
Photoaffinity analog of the semisynthetic echinocandin LY303366: identification of echinocandin targets in Candida albicans; Radding JA et al.; The echinocandins are a family of cyclic lipopeptides with potent antifungal activity . These compounds inhibit the synthesis of BETA-1,3-glucan in fungi . The new semisynthetic echinocandin LY303366 was derivatized to produce a photoactivatable cross-linking echinocandin analog with antifungal activity . This analog was radioiodinated and used as a probe in microsomal membrane preparations of Candida albicans which contain glucan synthase activity . The photoaffinity probe identified two major proteins of 40 and 18 kDa in both membrane preparations . Labeling of these proteins was specific in that it required irradiation with UV light and was effectively competed against with unlabeled echinocandin analogs . In addition, the abilities of echinocandin analogs to compete with the photoaffinity probe correlated to their relative antifungal potencies and glucan synthase inhibition . The 40-kDa protein was isolated, and partial sequences were obtained from internal peptide fragments of the protein . Analysis of the sequences of these internal peptides of the 40-kDa protein revealed that it was a new protein not previously described as being involved in glucan synthesis or the mode of action of echinocandins.

Antimicrob Agents Chemother, 1998 May, 42(5), 1160 - 7
Sequencing, disruption, and characterization of the Candida albicans sterol methyltransferase (ERG6) gene: drug susceptibility studies in erg6 mutants; Jensen-Pergakes KL et al.; The rise in the frequency of fungal infections and the increased resistance noted to the widely employed azole antifungals make the development of new antifungals imperative for human health . The sterol biosynthetic pathway has been exploited for the development of several antifungal agents (allylamines, morpholines, azoles), but additional potential sites for antifungal agent development are yet to be fully investigated . The sterol methyltransferase gene (ERG6) catalyzes a biosynthetic step not found in humans and has been shown to result in several compromised phenotypes, most notably markedly increased permeability, when disrupted in Saccharomyces cerevisiae . The Candida albicans ERG6 gene was isolated by complementation of a S . cerevisiae erg6 mutant by using a C . albicans genomic library . Sequencing of the Candida ERG6 gene revealed high homology with the Saccharomyces version of ERG6 . The first copy of the Candida ERG6 gene was disrupted by transforming with the URA3 blaster system, and the second copy was disrupted by both URA3 blaster transformation and mitotic recombination . The resulting erg6 strains were shown to be hypersusceptible to a number of sterol synthesis and metabolic inhibitors, including terbinafine, tridemorph, fenpropiomorph, fluphenazine, cycloheximide, cerulenin, and brefeldin A . No increase in susceptibility to azoles was noted . Inhibitors of the ERG6 gene product would make the cell increasingly susceptible to antifungal agents as well as to new agents which normally would be excluded and would allow for clinical treatment at lower dosages . In addition, the availability of ERG6 would allow for its use as a screen for new antifungals targeted specifically to the sterol methyltransferase.

Antimicrob Agents Chemother, 1998 May, 42(5), 1105 - 9
Pharmacodynamics of fluconazole in a murine model of systemic candidiasis; Louie A et al.; In this study we defined the pharmacodynamic parameter that optimizes outcome in deep-seated Candida albicans infections treated with fluconazole . Using a murine model of systemic candidiasis, we conducted single-dose dose-ranging studies with fluconazole to determine the dosage of this drug that resulted in a 50% reduction in fungal densities (50% effective dose {ED50}) in kidneys versus the fungal densities in the kidneys of untreated controls . We found that the ED50 of fluconazole given intraperitoneally was 4.56 mg/kg of body weight/day (95% confidence interval, 3.60 to 5.53 mg/kg/day), and the dose-response relationship was best described by an inhibitory sigmoid maximal effect (Emax) curve . To define the pharmacodynamics of fluconazole, we gave dosages lower than, approximating, and higher than the ED50 of fluconazole (range, 3.5 to 5.5 mg/kg/day, equivalent to the ED16 to the ED75) to various groups of infected animals using three dose-fractionation schedules . For each total dose of fluconazole examined, the dose-fractionation schedules optimized the ratio of the area under the concentration-time curve (AUC) to the MIC (the AUC/MIC ratio), the ratio of the maximum concentration of drug in serum (Cmax) to the MIC, and the time that the drug remained above the MIC for the infecting C . albicans isolate . Similar reductions in fungal densities in kidneys were seen between groups that received the same total dose of fluconazole in one, two, or four equally divided doses . Thus, dose-fractionation studies demonstrated that the pharmacodynamic parameter of fluconazole that best predicted outcome was the AUC/MIC ratio.

J Antibiot (Tokyo), 1997 Nov, 50(11), 937 - 43
In vitro antifungal activity of CAN-296: a naturally occurring complex carbohydrate; Ben-Josef AM et al.; The in vitro activity of a naturally occurring complex carbohydrate, CAN-296, was evaluated by testing 132 clinical and ATCC isolates of yeast and Aspergillus fumigatus, many of which were azole-resistant . The in vitro susceptibility tests were performed by standardized broth micro- and macrodilution methods and results were compared with those obtained for amphotericin B, fluconazole, ketoconazole, flucytosine and the pneumocandin L-733,560 . All tested Candida species showed highly uniform susceptibility to CAN-296 at concentrations of 0.078 to 0.312 microgram/ml; non-albicans Candida were as susceptible to CAN-296 as the Candida albicans strains . Multi-azole-resistant Candida species were highly sensitive to CAN-296 . Minimum inhibitory concentration measurements did not differ from minimum lethal concentrations by more than two-fold for all tested Candida species . Aspergillus fumigatus, on the other hand, showed only moderate susceptibility to CAN-296 . The kinetics of the anti-Candida activity of CAN-296 was investigated by kill-curve experiments using C . albicans and C . glabrata and the results were compared with those obtain for amphotericin B . CAN-296 was found to be rapidly fungicidal in concentrations ranging from 4-16 fold the mean MIC value . The broad spectrum of anti-Candida activity together with the rapid fungicidal effect make this complex carbohydrate a promising agent for clinical use.

Int J Prosthodont, 1998 Jan-Feb, 11(1), 75 - 81
Adherence of phenotypically switched Candida albicans to denture base materials; Radford DR et al.; PURPOSE: The purpose of this study was to investigate in vitro the levels of adhesion of phenotypically switched and nonswitched Candida albicans to the smooth and rough surfaces of heat-cured acrylic resin, Molloplast B, and Novus . MATERIALS AND METHODS: Nonswitched and switched cells of C . albicans 3153A were prepared and adhesion assays were performed . RESULTS: There was no difference in the level of adhesion of blastospores in their nonswitched or switched state . The adherence of hyphae to all three denture base materials was statistically significantly increased when in a switched form . There was greater adhesion to the two soft lining materials than to the acrylic resin . CONCLUSIONS: Rough surfaces promote adhesion . There was no significant difference in the adhesion of switched and nonswitched blastospores, but there was increased adhesion of hyphal cells.

Egypt Dent J, 1994 Jul, 40(3), 785 - 90
Effect of addition antimicrobial agents to denture reliners; el-Charkawi H et al.; This study aimed at evaluating the incorporation of two antimicrobial drugs (nystatin and polynoxylin) as regards: the effect of the liner on the activity of the drug, determination of the least effective concentration of each drug and its duration of action, as well as assessment of the effect of the drug on the mechanical properties and the chemical composition of the liner . Results showed that nystatin added to denture liners in three different concentrations by weight (3%, 5%, 10%) acted effectively against Candida albicans, and that there was a direct relationship between concentration of Nystatin and its duration of action . The inhibitory effect of nystatin (10%) lasted for at least 32 weeks (end of study period) . Furthermore, this concentration did not affect the strength properties of the liner . On the other hand, polynoxylin inhibited a number of strains of bacteria and Candida only in high concentrations (40-60%), and these concentrations adversely affected the strength properties of the liner.

Zentralbl Veterinarmed B, 1998 Apr, 45(3), 129 - 32
Relationship between cell counts in bovine milk and the presence of mastitis pathogens (yeasts and bacteria); Moretti A et al.; Bovine mastitis is an economically important disease in dairy industry and a variety of pathogen microorganisms are involved . The AA carried out a study to investigate the prevalence of yeasts and bacteria in milk samples (794) from dairy cows of Umbria (Central Italy), belonging to 19 herds, and the relationship between cell counts in bovine milk and the presence of these pathogens . 29.7% milk samples were positive for pathogen microorganisms, of those 4.9% were positive for yeasts and bacteria, 4.4% for yeasts and 20.4% for bacteria . The species of yeasts and bacteria most frequently encountered were Trichosporon capitatum (31.2%), T . beigelii (18.72%) and Candida albicans (12.48%), C . guillermondii (12.48%), C . tropicalis (12.48%); with regard to bacteria were Staphylococcus aureus (34.3%) and S . albo (19.8%) . The presence of yeasts and bacteria in milk samples are correlated to an increase of somatic cell counts even if with different degree . Epidemiological and sanitary correlation were carried out.

J Nat Prod, 1998 Apr, 61(4), 542 - 5
Novel bioactivities of Curcuma longa constituents; Roth GN et al.; Bioassay-directed fractionation of ethyl acetate extract from Curcuma longa Linn . rhizomes yielded three curcuminoids, which displayed topoisomerase I and II enzyme inhibition activity . Curcumin III (3) was the most active curcuminoid, inhibiting topoisomerase at 25 micrograms mL-1 . Curcumin I (1) and curcumin II (2) inhibited the topoisomerases at 50 micrograms mL-1 . Fractionation of the volatile oil from the rhizomes afforded ar-turmerone (4), which displayed mosquitocidal activity with an LD100 of 50 micrograms mL-1 on Aedes aegyptii larvae . Bioassay-directed fractionation of hexane extract from the turmeric leaves yielded labda-8(17),12-diene-15,16 dial (5) with antifungal activity against Candida albicans at 1 micrograms mL-1 and inhibited the growth of Candida kruseii and Candida parapsilosis at 25 micrograms mL-1 . In addition, 5 displayed 100% mosquitocidal activity on A . aegyptii larvae at 10 micrograms mL-1.

Aust Dent J, 1998 Feb, 43(1), 45 - 50
Candida-associated denture stomatitis . Aetiology and management: a review . Part 1 . Factors influencing distribution of Candida species in the oral cavity; Webb BC et al.; Candida species are yeasts and within the oral cavity, Candida albicans is the most frequently isolated . There is clear evidence that C . albicans adheres to oral surfaces including acrylic dentures and mucosa . The mechanisms of attachment differ, with candidal adhesion to inert surfaces under the control of hydrophobic and electrostatic forces and adhesion to mucosa dependent on a number of complex ligand-recognition systems . Other factors within the oral environment such as saliva, pH, bacteria and hyphal formation have been shown to influence adhesion of candida species to surfaces in the mouth.

Genitourin Med, 1997 Dec, 73(6), 475 - 6
Women with recurrent vaginal candidosis have normal peripheral blood B and T lymphocyte subset levels; White DJ et al.; OBJECTIVE: To compare the B and T lymphocyte subset levels of otherwise healthy women suffering from frequently recurrent vaginal candidosis with a healthy control group . SUBJECTS: 26 unselected otherwise healthy women of reproductive age with at least four attacks of vaginal candidosis in the past year and more than three vaginal isolates of a moderate or heavy growth of Candida albicans . Controls were 26 patients or clinical and laboratory staff (asymptomatic for genital infection) matched for time of day and age within 5 years . Only three patients accepted an HIV test . All proved HIV negative . No controls were tested . MAIN OUTCOME MEASURES: T lymphocyte subsets (CD4 and 8) and B lymphocytes (CD 19) as estimated from the total lymphocyte count and flow cytometry . RESULTS: No statistically significant difference between patients and controls . CONCLUSION: No significant difference was found between patients and controls in levels of lymphocyte subsets.

Int J Dermatol, 1998 Feb, 37(2), 145 - 9
Epidemiology and in vitro activity of antimycotics against candidal vaginal/skin/nail infections in Singapore; Kwok YK et al.; BACKGROUND: Candidal infections of the skin/nails and vagina are very common worldwide . Various in vitro test systems are available to help to determine the antifungal activity of drugs . The minimum inhibitory concentration (MIC) is a standard measure of the in vitro potency of drugs against yeasts . METHODS: Vaginal smears and skin/nail scrapings of 50 consecutive patients with candidal vaginitis and 46 consecutive patients (28 women, 18 men) with cutaneous/nail candidosis were used in the study . Direct microscopy and culture from vaginal smears and skin scrapings were performed on all patients . The MICs were determined using the broth dilution method . RESULTS: For vaginal candidosis, the mean age of the patients was 28.2 years (range, 9-49 years) . Candida albicans accounted for 58% of the isolates, C . glabrata for 32%, C . tropicalis for 6%, and C . parasilosis for 4% . At the MIC of < or = 4 mg/L, 65-95% of C . albicans, 66-94% of C . glabrata, 33-100% of C . tropicalis, and 0-50% of C . parasilosis were susceptible to the drugs tested (ketoconazole, itraconazole, nystatin, amorolfine, clotrimazole, and miconazole) . For cutaneous/nail candidosis, the mean age of the patients was 45 years (range, 19-82 years) . C . albicans made up 59% of the isolates, C . parasilosis 20%, C . krusei 13%, C . glabrata 4%, and C . tropicalis 4% . At the MIC of < or = 4 mg/L, 59-96% of C . albicans, 100% of C . glabrata, 83-100% of C . krusei, 89-100% of C . parasilosis, and 100% of C . tropicalis were susceptible to the drugs tested (ketoconazole, itraconazole, nystatin, amorolfine, clotrimazole, and miconazole) . CONCLUSIONS: C . albicans is the most common Candida species causing cutaneous/nail and vaginal candidosis in Singapore . The in vitro antifungal activities of ketoconazole, itraconazole, nystatin, amorolfine, clotrimazole, and miconazole are similar against the various Candida species . C . parasilosis in vaginal candidosis appears to be less susceptible . Here, itraconazole and amorolfine may be more effective.

Glycobiology, 1998 Mar, 8(3), 221 - 5
Differential binding of lectins IL-2 and CSL to candida albicans and cancer cells; Zanetta JP et al.; The demonstration that interleukin 2 (IL-2) is a lectin specific for oligomannosides allows to understand a new function for this cytokine: as a bifunctional molecule when bound to its receptor ss, IL-2 associates the latter which the CD3/TCR complex, interacting with oligosaccharides of CD3 through its carbohydrate-recognition domain (Zanetta et al . , 1996, Biochem . J., 318, 49-53) . This induces the tyrosine phosphorylation of the IL-2R beta by ++p56(lck) , the first step of the IL-2-dependent signaling . Since this specific association is disrupted in vitro by oligomannosides with five and six mannose residues, we made the hypothesis that pathogenic cells or microorganisms could bind IL-2, consequently disturbing the IL-2-dependent response . This study shows that the pathogenic yeast Candida albicans (in contrast with nonpathogenic yeasts) binds high amounts of IL-2 as did cancer cells . In contrast with cancer cells, yeasts do not bind the Man6GlcNAc2-specific lectin CSL, an endogenous "amplifier of activation signals" (Zanetta et al . , 1995, Biochem . J., 311, 629-636).

Nippon Ishinkin Gakkai Zasshi, 1998, 39(2), 103 - 7
{Anti-Candida activities of azole antifungals in the presence of lysozyme in vitro}; Abe S et al.; The combined effects of lysozyme and various antifungal agents were investigated by microbroth dilution method against Candida albicans in vitro . Synergistic anti-Candida activity was observed between egg white lysozyme and itraconazole, clotrimazole, miconazole and lanoconazole . Similar, although not as clear, combination anti-Candida activities were seen in the cases of ketoconazole, bifonazole, amphotericin B and nystatin . Anti-Candida activity of fluconazole was not affected by the addition of lysozyme, however . Physiological roles of this combination effect in anti-Candida therapy by azole antifungals were discussed.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(2), 79 - 83
{Molecular cloning of Candida albicans phospholipase D}; Kanoh H et al.; Phospholipase D (PLD) catalyses the hydrolysis of phosphatidylcholine, a major substrate, to phosphatidic acid and choline, and its activity is regulated by a variety of hormones, growth factors, and other extracellular signals in mammalian cells . Thus, it is now recognized as a signal transducing enzyme such as phosphatidylinositol-specific phospholipase C, adenylate cyclase, or protein tyrosine kinases . Furthermore, recent findings that regulation by members of the ADP-ribosylation factor (ARF) and Rho families of monomeric GTP-binding protein suggest roles of PLD in intracellular vesicle traffi-cking, morphological changes, and mitogenic signaling process . In Saccharomyces cerevisiae, PLD gene has been cloned and revealed to be essential for meiosis . In contrast, little is known about PLD in Candida albicans . As a first step to understand possible physiological roles of PLD in C . albicans, we cloned a PLD gene from a C . albicans genomic DNA library . Deduced amino acid sequence analysis showed the structural similarity to mammalian, yeast, and plant PLDs . It was also suggested employing RT-PCR (reverse transcriptase polymerase chain reaction) that an isozyme of C . albicans PLD was present.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(2), 73 - 8
Drug pumping mechanisms in Candida albicans; Cannon RD et al.; Multiple drug resistance is becoming a major problem in the treatment of AIDS patients with oropharyngeal candidosis . Candida albicans strains isolated from candidosis patients who do not respond to fluconazole therapy often show azole drug resistance which usually correlates with the expression of C . albicans CDR1, CDR2 or BENr genes, encoding potential drug efflux pumps . The objective of this study was to develop a yeast secretory vesicle transport assay and use this system to study the pumping function of Cdr1 and Benr . The C . albicans CDR1 and BEN r genes were cloned separately into plasmid pVT101-U, to form plasmids pKY1011 and pKN5001 respectively . Plasmids pVT101-U, pKY1011 and pKN5001 were transformed into Saccharomyces cerevisiae SY1, a sec6-4 mutant with a temperature-sensitive mutation in the secretory pathway . SY1 cells transformed with pKY1011 or pKN5001, were more resistant to fluconazole (MICs in both cases 64 microg/ml) than SY1 cells (MIC 32 microg/ml) . In addition, cells transformed with pKY1011 were more resistant to cycloheximide (MIC 16 microg/ml) than SY1 cells (MIC 2 microg/ml) . Intact secretory vesicles were isolated from SY1 cells expressing Cdr1 and these vesicles accumulated fluconazole in a time dependent manner . These experiments demonstrated that S . cerevisiae secretory vesicles can be used to examine the mechanism of fluconazole transport by putative C . albicans membrane pumps.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(2), 67 - 71
{Rbf1 (RPG-box binding factor), a transcription factor involved in yeast-hyphal transition of Candida albicans}; Aoki Y et al.; The major fungal pathogen for fungal diseases which have become a major medical problem in the last few years is Candida albicans, which can grow both in yeast and hyphae forms . This ability of C . albicans is thought to contribute to its colonization and dissemination within host tissues . In a recent few years, accompanying the introduction of molecular biological tools into C . albicans organism, several factors involved in the signal transduction pathway for yeast-hyphal transition have been identified . One MAP kinase pathway in C . albicans, similar to that leading to STE12 activation in Saccharomyces cerevisiae, has been reported . C . albicans strains mutant in these genes show retarded filamentous growth on a solid media but no impairment of filamentous growth in mice . These results suggest two scenarios that a kinase signaling cascade plays a part in stimulating the morphological transition in C . albicans, and that there would be another signaling pathway effective in animals . In this latter true hyphal pathway, although some candidate proteins, such as Efg1 (transcription factor), Int1 (integrin-like membrane protein), or Phr1 (pH-regulated membrane protein), have been identified, it is still too early to say that we understand the whole picture of that cascade . We have cloned a C . albicans gene encoding a novel DNA binding protein, Rbf1, that predominantly localizes in the nucleus, and shows transcriptional activation capability . Disruption of the functional RBF1 genes of C . albicans induced the filamentous growth on all solid and liquid media tested, suggesting that Rbf1 might be another candidate for the true hyphal pathway . Relationships with other factors described above, and the target (regulated) genes of Rbf1 is under investigation.

J Invest Dermatol, 1998 May, 110(5), 811 - 7
Aloe barbadensis extracts reduce the production of interleukin-10 after exposure to ultraviolet radiation; Byeon SW et al.; Cutaneous exposure to ultraviolet radiation suppresses the induction of T cell mediated responses such as contact and delayed type hypersensitivity (DTH) by altering the function of immune cells in the skin and causing the release of immunoregulatory cytokines . Extracts of crude Aloe barbadensis gel prevent this photosuppression . Because the regulation of contact hypersensitivity and DTH responses differ, we investigated whether protection was afforded by a single or multiple agents in Aloe and the mechanism by which this material prevents suppression of DTH immunity . The ability of Aloe gel to prevent suppression of contact hypersensitivity responses to hapten decayed rapidly after manufacture . In contrast, agents that protected against systemic suppression of DTH responses to Candida albicans were stable over time . Oligosaccharides prepared from purified Aloe polysaccharide prevented suppression of DTH responses in vivo and reduced the amount of IL-10 observed in ultraviolet irradiated murine epidermis . To assess the effect of Aloe extracts on keratinocytes, Pam 212 cells were exposed in vitro to ultraviolet radiation and treated for 1 h with Aloe oligosaccharides . Culture supernatants were collected 24 h later and injected into mice . Supernatants from ultraviolet irradiated keratinocytes suppressed the induction of DTH responses, whereas Aloe oligosaccharide treatment reduced IL-10 and blocked the suppressive activity of the supernatants . These results indicate that Aloe contains multiple immunoprotective factors and that Aloe oligosaccharides may prevent ultraviolet induced suppression of DTH by reducing keratinocyte derived immunosuppressive cytokines.

Microbiology, 1998 Apr, 144 ( Pt 4), 849 - 57
Repetitive sequences (RPSs) in the chromosomes of Candida albicans are sandwiched between two novel stretches, HOK and RB2, common to each chromosome; Chindamporn A et al.; A novel sequence designated HOK, which is next to the RPS, a repetitive sequence specific to Candida albicans, was cloned and sequenced . HOK hybridized with all of the chromosomes on which the RPSs were located, but did not hybridize with chromosome 3, which does not harbour any RPSs . Sequence determination revealed that a portion of HOK has significant homology with the B and C1 fragments of Ca3, which is used as a molecular epidemiological probe . A homology search of the deduced amino acids of HOK against the protein database showed partial homology with an isocitrate dehydrogenase of Saccharomyces cerevisiae, although an ORF large enough to encode the enzyme was not detected . To verify the existence of other sequences homologous with HOK, a portion of the HOK sequence was amplified using PCR . Sequence determination of the 41 clones from the PCR products resulted in at least six HOK-homologous clones . Another RPS-containing clone, RB2, was isolated from the Pstl-digested chromosome R or 1 . It was determined that RB2a, one of the subclones from RB2, hybridized with all of the chromosomes, including chromosome 3, with which neither HOK nor RPS hybridized . The hybridization profile also showed that RPS is located between HOK and RB2a on chromosomes other than chromosome 3.

Microbiology, 1998 Apr, 144 ( Pt 4), 839 - 47
A Candida albicans 37 kDa polypeptide with homology to the laminin receptor is a component of the translational machinery; Montero M et al.; A cDNA encoding a 37 kDa protein was isolated from an expression library using antibodies raised against mycelial cell walls from Candida albicans . The 37 kDa protein has over 60% sequence identity with the 37 kDa laminin-binding protein (LBP) from humans and over 80% identity with the Yst proteins of Saccharomyces cerevisiae . The C . albicans protein was named CaYst1 . It was found in membrane and ribosome fractions but surprisingly, was not found in cell walls . Unlike the human LBP, CaYst1p does not bind laminin . These data indicate that CaYst1p is not a cell-surface receptor for laminin as has been proposed for the human LBP . Instead, like the S . cerevisiae Yst proteins, it appears to be a ribosomal protein . This conclusion is supported by the finding that CaYST1-cDNA complements the lethal phenotype linked to the disruption of both YST genes in S . cerevisiae.

Microbiology, 1998 Apr, 144 ( Pt 4), 829 - 38
Candida dubliniensis: phylogeny and putative virulence factors; Gilfillan GD et al.; Candida dubliniensis is a recently identified species which is implicated in oral candidosis in HIV-infected and AIDS patients . The species shares many phenotypic characteristics with, and is phylogenetically closely related to, Candida albicans . In this study the phylogenetic relationship between these two species was investigated and a comparison of putative virulence factors was performed . Four isolates of C . dubliniensis from different clinical sources were chosen for comparison with two reference C . albicans strains . First, the distinct phylogenetic position of C . dubliniensis was further established by the comparison of the sequence of its small rRNA subunit with representative Candida species . The C . dubliniensis isolates formed true unconstricted hyphae under most induction conditions tested but failed to produce true hyphae when induced using N-acetylglucosamine . Oral C . dubliniensis isolates were more adherent to human buccal epithelial cells than the reference C . albicans isolates when grown in glucose and equally adherent when grown in galactose . The C . dubliniensis isolates were sensitive to fluconazole, itraconazole, ketoconazole and amphotericin B . Homologues of seven tested C . albicans secretory aspartyl proteinase (SAP) genes were detected in C . dubliniensis by Southern analysis . In vivo virulence assays using a systemic mouse model suggest that C . dubliniensis is marginally less virulent than C . albicans . These data further confirm the distinct phenotypic and genotypic nature of C . dubliniensis and suggest that this species may be particularly adapted to colonization of the oral cavity.

J Antimicrob Chemother, 1998 Mar, 41(3), 357 - 66
The effect of antifungal drugs in combination on the growth of Candida glabrata in solid and liquid media; Siau H et al.; The effect of drugs in combination on the growth of Candida glabrata was studied in solid medium by demonstration of reduced or enhanced growth, and in liquid medium by determination of interaction indices . Amphotericin B and 5-fluorocytosine showed a synergic effect, while combinations of amphotericin B and miconazole, and miconazole and 5-fluorocytosine exhibited antagonistic effects . In addition, concentration-dependent antagonism was observed amongst combinations of metabolic inhibitors, between inhibitors of sterol biosynthesis (terbinafine, miconazole, ketoconazole, clotrimazole, econazole, fluconazole, itraconazole and amorolfine) and inhibitors of nucleic acid or protein biosynthesis (5-fluorocytosine, 5-fluorouracil, rifampicin and chlortetracycline) . This antagonism was strain-specific, occurring with C . glabrata strain 4, Saccharomyces cerevisiae strain 237 and Candida albicans strain 72R, but not with C . albicans strain 6406 or Candida parapsilosis strain 3104.

Prostaglandins Leukot Essent Fatty Acids, 1998 Feb, 58(2), 85 - 90
Role of arachidonic acid metabolites in the action of a beta adrenergic agonist on human monocyte phagocytosis; Borda ES et al.; The mechanisms by which beta adrenergic stimulation regulates phagocytosis of Candida albicans by human peripheral monocytes (HPM) are characterized . Isoproterenol (ISO) inhibits phagocytosis in a concentration-dependent manner . This effect was blunted by propranolol, inhibitors of phospholipase A2 (PLA2), cyclooxygenase and verapamil, pointing to a participation of arachidonic acid (AA) metabolites and calcium in the phenomenon . Prostaglandin E2 (PGE2) and dibutyryl cyclic AMP (db-cAMP) also exerted the same inhibitory effect on phagocytosis . ISO interacts with beta adrenergic receptors of HPM increasing PGE2 and cAMP . We conclude that the mechanisms by which beta adrenergic stimulation regulates phagocytosis of Candida albicans by HPM appear to be secondary to beta adrenoceptor-mediated hydrolysis of AA accompanied by an increase in PGE2 generation and cAMP production . Both PGE2 and cAMP could act as mediators of the inhibitory action of beta agonists on the HPM-phagocytosis process.

J Oral Rehabil, 1998 Feb, 25(2), 135 - 8
In vivo and in vitro study of fungal presence and growth on three tissue conditioning materials on implant supported complete denture wearers; Kulak Y et al.; In this study three type of tissue conditioning materials were used in vitro and in vivo to investigate the presence and growth of Candida albicans, considered to be the pathogenic member of the genus . In vitro test results of different tissue conditioners showed that they have a tendency to have an inhibiting effect on C . albicans at the third day of incubation . For the in vivo tests tissue conditioners were placed in existing maxillary prosthesis of 21 patients who had been treated with endosseous dental implants (seven in each group) . The results showed that yeast forms were observed after 3 days in two patients' dentures which were relined with Fitt . Yeasts forms were also seen in three patients' dentures after 6 days, two of them relined with Fixo-gel and the other one relined with Visco-gel . The hyphal form of Candida was seen in four patients' dentures, relined with Fitt, and also in three patients' dentures, relined with Fixo-gel and Visco-gel.

J Prosthet Dent, 1998 Apr, 79(4), 454 - 8
Use of microwave energy to disinfect a long-term soft lining material contaminated with Candida albicans or Staphylococcus aureus; Baysan A et al.; STATEMENT OF PROBLEM: Soft lining materials have been found to be more susceptible to microbial adhesion than acrylic resin base materials . Denture hygiene is essential to maintain the serviceability of the denture, and microwave energy has been suggested for denture disinfection . PURPOSE: The purpose of this study was to determine the effectiveness of microwave energy in the disinfection of a long-term soft lining material . MATERIAL AND METHODS: A long-term soft lining material was contaminated with known microorganisms and the reduction of organism counts after test disinfection regimes calculated . The disinfection regimes were microwaving for 5 minutes, leaving dry overnight, and soaking overnight in a dilute sodium hypochlorite solution . The test microorganisms were Candida albicans or Staphylococcus aureus . RESULTS: For both organisms, soaking in sodium hypochlorite reduced the number of viable adherent microorganisms recovered significantly more than exposure to microwave energy, which led to greater reduction than leaving the lining material dry overnight (p < 0.001, Wilcoxon nonparametric signed rank test) . CONCLUSION: With reference to the tested microorganisms, disinfection of Molloplast-b soft lining material in dilute sodium hypochlorite solution proved to be more effective than exposure to microwave energy, which in turn was more effective than leaving the lining dry overnight.

Infect Immun, 1998 May, 66(5), 2078 - 84
Cloning and characterization of CAD1/AAF1, a gene from Candida albicans that induces adherence to endothelial cells after expression in Saccharomyces cerevisiae; Fu Y et al.; Adherence to the endothelial cell lining of the vasculature is probably a critical step in the egress of Candida albicans from the intravascular compartment . To identify potential adhesins that mediate the attachment of this organism to endothelial cells, a genomic library from C . albicans was used to transform a nonadherent strain of Saccharomyces cerevisiae . The population of transformed yeasts was enriched for highly adherent clones by repeated passages over endothelial cells . One clone which exhibited a fivefold increase in endothelial cell adherence, compared with S . cerevisiae transformed with vector alone, was identified . This organism also flocculated . The candidal DNA fragment within this adherent/flocculent organism was found to contain a single 1.8-kb open reading frame, which was designated CAD1 . It was found to be identical to AAF1 . The predicted protein encoded by CAD1/AAF1 contained features suggestive of a regulatory factor . Consistent with this finding, immunoelectron microscopy revealed that CAD1/AAF1 localized to the cytoplasm and nucleus but not the cell wall or plasma membrane of the transformed yeasts . Because yeasts transformed with CAD1/AAF1 both flocculated and exhibited increased endothelial cell adherence, the relationship between adherence and flocculation was examined . S . cerevisiae expressing either of two flocculation phenotypes, Flo1 or NewFlo, adhered to endothelial cells as avidly as did yeasts expressing CAD1/AAF1 . Inhibition studies revealed that the flocculation phenotype induced by CAD1/AAF1 was similar to Flo1 . Thus, CAD1/AAF1 probably encodes a regulatory protein that stimulates endothelial cell adherence in S . cerevisiae by inducing a flocculation phenotype . Whether CAD1/AAF1 contributes to the adherence of C . albicans to endothelial cells remains to be determined.

Allergy, 1998 Apr, 53(4), 359 - 66
In-house reference (IHR) preparation of Candida albicans allergen extract . A standardized extraction procedure; Savolainen J et al.; A standardized, controlled procedure for preparation of an in-house reference (IHR) preparation of an allergen extract of Candida albicans is described . The procedure, based on previous studies of allergens of C . albicans, is designed to yield a maximum of allergens in optimum extraction conditions and to provide a reference preparation for further extract production . The SDS-PAGE, IgE-immunoblotting, and crossed radioimmunoelectrophoresis (CRIE) analyses showed that the procedure is reproducible with acceptable batch-to-batch variation . The variation in the content of the most important allergens, namely, proteins with molecular weights of 46, 29, and 27 kDa in the pooled final batches, is acceptable (coeff . of variation < 15%), although in the intermediate batches of different strains, the coefficient of variation may occasionally exceed 20% . A comparison with other C . albicans allergen preparations used in our previous studies is also presented . The resulting extract can be used as a reference in further extract production and also in experimental in vitro and in vivo studies.

J Clin Microbiol, 1998 May, 36(5), 1255 - 9
Nonperinatal nosocomial transmission of Candida albicans in a neonatal intensive care unit: prospective study; Reef SE et al.; Nosocomial Candida albicans infections have become a major cause of morbidity and mortality in neonates in neonatal intensive care units (NICUs) . To determine the possible modes of acquisition of C . albicans in hospitalized neonates, we conducted a prospective study at Grady Memorial Hospital, Atlanta, Ga . Clinical samples for fungal surveillance cultures were obtained at birth from infants (mouth, umbilicus, and groin) and their mothers (mouth and vagina) and were obtained from infants weekly until they were discharged . All infants were culture negative for C . albicans at birth . Six infants acquired C . albicans during their NICU stay . Thirty-four (53%) of 64 mothers were C . albicans positive (positive at the mouth, n = 26; positive at the vagina, n = 18; positive at both sites, n = 10) at the time of the infant's delivery . A total of 49 C . albicans isolates were analyzed by restriction endonuclease analysis and restriction fragment length polymorphism analysis by using genomic blots hybridized with the CARE-2 probe . Of the mothers positive for C . albicans, 3 of 10 were colonized with identical strains at two different body sites, whereas 7 of 10 harbored nonidentical strains at the two different body sites . Four of six infants who acquired C . albicans colonization in the NICU had C . albicans-positive mothers; specimens from all mother-infant pairs had different restriction endonuclease and CARE-2 hybridization profiles . One C . albicans-colonized infant developed candidemia; the colonizing and infecting strains had identical banding patterns . Our study indicates that nonperinatal nosocomial transmission of C . albicans is the predominant mode of acquisition by neonates in NICUs at this hospital; mothers may be colonized with multiple strains of C . albicans simultaneously; colonizing C . albicans strains can cause invasive disease in neonates; and molecular biology-based techniques are necessary to determine the epidemiologic relatedness of maternal and infant C . albicans isolates and to facilitate determination of the mode of transmission.

Oral Microbiol Immunol, 1997 Dec, 12(6), 358 - 65
Phenotypic and genotypic characterization of oral yeasts from Finland and the United States; Hannula J et al.; A total of 4-22 isolates of oral yeasts per subjects from 48 yeast-positive Finnish and American subjects (25 females and 23 males) were phenotyped and genotyped to determine the frequency of simultaneous oral carriage of multiple yeast taxa . An oral sample from either periodontal pockets, oral mucosa or saliva was obtained . All subjects yielded Candida albicans and 3 subjects an additional yeast species (Candida krusei, Candida glabrata or Saccharomyces cerevisiae) . The API 20C Aux kit distinguished 9 different carbohydrate assimilation profiles among the C . albicans isolates . Thirty-eight of 46 C . albicans biotype I isolates were categorized in a single numerical profile . PCR analysis, using a random primer OPA-03 and a repetitive primer (GACA)4, detected 2 major genotypic groups among the C . albicans isolates; 44 subjects showing isolates with a "typical" PCR-profile and 4 subjects isolates with an "atypical" PCR-profile . The "atypical" PCR-profile was similar to that of Candida dubliniensis . All C . albicans isolates assimilated xylose, except 5, including the 4 with an "atypical" PCR-profile . No difference was found in distribution of oral yeast species, and of C . albicans phenotypes and genotypes between Finnish and American subjects . The present PCR method may offer a rapid and easy means of distinguishing oral Candida species.

Infect Immun, 1998 May, 66(5), 2154 - 62
A 70-kilodalton recombinant heat shock protein of Candida albicans is highly immunogenic and enhances systemic murine candidiasis; Bromuro C et al.; The 70-kDa recombinant Candida albicans heat shock protein (CaHsp70) and its 21-kDa C-terminal and 28-kDa N-terminal fragments (CaHsp70-Cter and CaHsp70-Nter, respectively) were studied for their immunogenicity, including proinflammatory cytokine induction in vitro and in vivo, and protection in a murine model of hematogenous candidiasis . The whole protein and its two fragments were strong inducers of both antibody (Ab; immunoglobulin G1 {IgG1} and IgG2b were the prevalent isotypes) and cell-mediated immunity (CMI) responses in mice . CaHsp70 preparations were also recognized as CMI targets by peripheral blood mononuclear cells of healthy human subjects . Inoculation of CaHsp70 preparations into immunized mice induced rapid production of interleukin-6 (IL-6) and tumor necrosis factor alpha, peaking at 2 to 5 h and declining within 24 h . CaHsp70 and CaHsp70-Cter also induced gamma interferon (IFN-gamma), IL-12, and IL-10 but not IL-4 production by CD4+ lymphocytes cocultured with splenic accessory cells from nonimmunized mice . In particular, the production of IFN-gamma was equal if not superior to that induced in the same cells by whole, heat-inactivated fungal cells or the mitogenic lectin concanavalin A . In immunized mice, however, IL-4 but not IL-12 was produced in addition to IFN-gamma upon in vitro stimulation of CD4+ cells with CaHsp70 and CaHsp70-Cter . These animals showed a decreased median survival time compared to nonimmunized mice, and their mortality was strictly associated with organ invasion by fungal hyphae . Their enhanced susceptibility was attributable to the immunization state, as it did not occur in congenitally athymic nude mice, which were unable to raise either Ab or CMI responses to CaHsp70 preparations . Together, our data demonstrate the elevated immunogenicity of CaHsp70, with which, however, no protection against but rather some enhancement of Candida infection seemed to occur in the mouse model used.

Infect Immun, 1998 May, 66(5), 2052 - 9
The cell wall-associated glyceraldehyde-3-phosphate dehydrogenase of Candida albicans is also a fibronectin and laminin binding protein; Gozalbo D et al.; By immunoelectron microscopy with a polyclonal antibody against the cytosolic glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Candida albicans (anti-GAPDH PAb), the protein was clearly detected at the outer surface of the cell wall, particularly on blastoconidia, as well as in the cytoplasm . Intact blastoconidia were able to adhere to fibronectin and laminin immobilized on microtiter plates, and this adhesion was markedly reduced by both the anti-GAPDH PAb and soluble GAPDH from Saccharomyces cerevisiae . In addition, semiquantitative flow cytometry analysis with the anti-GAPDH PAb showed a decrease in antibody binding to cells in the presence of soluble fibronectin and laminin . Purified cytosolic C . albicans GAPDH was found to bind to fibronectin and laminin in a ligand Western blot assay . These observations suggest that the cell wall-associated form of the GAPDH in C . albicans could be involved in mediating adhesion of fungal cells to fibronectin and laminin, thus contributing to the attachment of the microorganism to host tissues and to the dissemination of Candida infection.

Infect Immun, 1998 May, 66(5), 1953 - 61
Cloning and sequencing of a Candida albicans catalase gene and effects of disruption of this gene; Wysong DR et al.; Catalase plays a key role as an antioxidant, protecting aerobic organisms from the toxic effects of hydrogen peroxide, and in some cases has been postulated to be a virulence factor . To help elucidate the function of catalase in Candida albicans, a single C . albicans-derived catalase gene, designated CAT1, was isolated and cloned . Degenerate PCR primers based on highly conserved areas of other fungal catalase genes were used to amplify a 411-bp product from genomic DNA of C . albicans ATCC 10261 . By using this product as a probe, catalase clones were isolated from genomic libraries of C . albicans . Nucleotide sequence analysis revealed an open reading frame encoding a protein of 487 amino acid residues . Construction of a CAT1-deficient mutant was achieved by using the Ura-blaster technique for sequential disruption of multiple alleles by integrative transformation using URA3 as a selectable marker . Resulting mutants exhibited normal morphology and comparable growth rates of both yeast and mycelial forms . Enzymatic analysis revealed an abundance of catalase in the wild-type strain but decreasing catalase activity in heterozygous mutants and no detectable catalase in a homozygous null mutant . In vitro assays showed the mutant strains to be more sensitive to damage by both neutrophils and concentrations of exogenous peroxide that were sublethal for the parental strain . Compared to the parental strain, the homozygous null mutant strain was far less virulent for mice in an intravenous infection model of disseminated candidiasis . Definitive linkage of CAT1 with virulence would require restoration of activity by reintroduction of the gene into mutants . However, initial results in mice, taken together with the enhanced susceptibility of catalase-deficient hyphae to damage by human neutrophils, suggest that catalase may enhance the pathogenicity of C . albicans.

Infect Immun, 1998 May, 66(5), 1904 - 9
Hemoglobin-induced binding of Candida albicans to the cell-binding domain of fibronectin is independent of the Arg-Gly-Asp sequence; Yan S et al.; Hemoglobin specifically induces fibronectin (FN) binding to the pathogenic yeast Candida albicans . When grown in the complex medium Sabouraud broth, C . albicans expresses receptors that bind to several domains of FN . Growth in defined medium supplemented with 0.1% hemoglobin, however, enhanced the binding of FN to a single class of receptors, with a Kd = 4.6 x 10(-8) M . Competitive binding assays using recombinant and proteolytic fragments of FN revealed that the cell-binding domain mediated this interaction . A recombinant 40-kDa fragment of FN consisting of type III repeats 9 to 13 had an inhibitory activity similar to that of the entire 120-kDa cell-binding domain, indicating that the C-terminal portion of the cell-binding domain contains the binding site . A recombinant 33-kDa fragment of the cell-binding domain and a 33-kDa fragment with the RGD sequence deleted had the same inhibitory activities, demonstrating that the RGD sequence recognized by some mammalian integrins is not required . The addition of hemoglobin to the culture medium also enhanced Candida cell adhesion to immobilized FN and to 120- and 40-kDa fragments of FN but not to the collagen-binding or fibrin I domains . Using ligand protection, we identified a surface protein from C . albicans with an apparent molecular mass of 55 kDa that was protected by both FN and the 40-kDa fragment derived from the cell-binding domain . Therefore, hemoglobin both induces FN binding and changes the relative affinities of C . albicans for the cell- and collagen-binding domains of FN.

Phytochemistry, 1998 Mar, 47(5), 729 - 34
Antifungal and larvicidal meroterpenoid naphthoquinones and a naphthoxirene from the roots of Cordia linnaei; Ioset JR et al.; Three new meroterpenoid naphthoquinones, the known cordiaquinone B and a new naphthoxirene have been isolated from the roots of Cordia linnaei . Their structures were established by spectrometric methods including EI, D/CI and FAB mass spectrometry, 1H, 13C and 2D NMR experiments . The naphthoquinones showed activity against Cladosporium cucumerinum, Candida albicans and the larvae of the yellow fever-transmitting mosquito Aedes aegypti, while the naphthoxirene derivative was found to be inactive in the same bioassays.

Mol Cells, 1998 Feb 28, 8(1), 68 - 74
Characterization and intracellular localization of the Rok1 protein involved in yeast cell division; Rhee JY et al.; The ROK1 gene is essential for the cell cycle progression in Saccharomyces cerevisiae . ROK1 has been predicted to encode an ATP-dependent RNA helicase of the DEAD-box family . We have analyzed the ROK1 gene expression both at the protein and RNA levels . Polyclonal antibodies were raised against trpE::rok1 hybrid proteins and were affinity purified by using lacZ::rok1 hybrid proteins . Western blot experiments using anti-Rok1 antibodies revealed a single protein band of 64 kDa which is an expected size from the Rok1 amino acid sequence . Indirect immuno-fluorescence showed that the Rok1 protein is localized predominantly to the cytoplasm of the vegetatively growing cells . We have detected immunocross-reactive homologs of Rok1p in Candida albicans and Drosophila melanogaster.

Curr Opin Ophthalmol, 1998 Jun, 9(3), 66 - 70
Treatment outcomes of endogenous fungal endophthalmitis; Smiddy WE; Endogenous fungal endophthalmitis may present in a debilitated or otherwise healthy host, may be increasing in prevalence, and is most commonly caused by Candida albicans . Recognizing the potential for complications with systemic amphotericin B use, recent investigators have developed several newer principles governing the treatment of such cases . For cases of simple choroiditis or very minimal endophthalmitis (i.e., vitritis), systemic treatment with oral fluconazole may be effective; however, if vitritis symptoms persist or progress, vitrectomy allows for better clearing of the organism . Intravitreal amphotericin B in conjunction with vitrectomy has been advocated by many . The most recent series have shown that an extended course of oral fluconazole following vitrectomy without intravitreal amphotericin B affects resolution of infection in the vast majority of patients . Final visual acuity outcomes depend most on the site of initial choroiditis . If the macula is spared and preretinal membranes can be effectively removed, visual acuity results can be exceedingly good.

Microbiol Immunol, 1998, 42(3), 227 - 30
Discrimination among the clinical isolates of Candida albicans by amplification of the repetitive sequences, alts; Doi M et al.; A primer pair, PB and BSH, which amplified alts, a portion of Candida albicans-specific repetitive sequence, RPS, gave stable and reproducible fingerprint patterns of the strains by polymerase chain reaction (PCR) . We applied this method to clinical isolates of C . albicans for strain discrimination . Using PCR fingerprint patterns, we could analyze the relatedness of C . albicans strains including those isolated from children with leukemia and their bedside parents . The results indicated that PCR analysis targeting an alt region gives rise to the same conclusion as the previous study obtained by SmaI RFLP analysis.

J Marmara Univ Dent Fac, 1997 Sep, 2(4), 682 - 4
In vitro study of fungal presence and growth on three tissue conditioner materials; Kulak Y et al.; In this study three type of tissue conditioning materials (Visco-gel, Fixo-gel, Fitt) are used in vitro to investigate the presence and growth of Candida albicans . In vitro test results of different tissue conditioners showed no inhibiting effect on Candida albicans.

J Med Microbiol, 1998 Apr, 47(4), 359 - 63
Effect of granulocyte-macrophage colony-stimulating factor on candidacidal activity of neutrophils, monocytes or monocyte-derived macrophages and synergy with fluconazole; Natarajan U et al.; The effect of in-vitro granulocyte-macrophage colony-stimulating factor (GM-CSF) treatment of neutrophils, monocytes or monocyte-derived macrophages (MDM) on candidacidal activity was tested . Synergy of effector cells with fluconazole (FCZ) for enhanced killing was also investigated . Incubation of neutrophils with GM-CSF 0.67 microg/L plus Candida albicans Sh27 for 24 h significantly increased candidacidal activity (36% versus 74%) . Synergy with FCZ for killing was also significantly increased from 93% to 97% when neutrophils were incubated with GM-CSF . Monocytes cultured with GM-CSF and C . albicans for 24 h had significantly increased fungistatic activity compared to controls and synergy with FCZ for killing was significantly enhanced from 40% to 59% . Monocytes cultured for 3 days with GM-CSF had increased fungistatic activity compared to control MDM and showed synergy with FCZ for significantly enhanced killing (46%) compared to control MDM (12%).

Diabetes, 1998 Apr, 47(4), 566 - 9
The relationship between humoral and cellular immunity to IA-2 in IDDM; Ellis TM et al.; Autoantibodies to the neuroendocrine protein insulinoma-associated protein 2 (IA-2), a member of the tyrosine phosphatase family, have been observed in individuals with or at increased risk for IDDM . Because this disease is thought to result from a T-cell-mediated autoimmune destruction of the insulin-producing pancreatic beta-cells, we analyzed humoral and cellular immune reactivity to this autoantigen to further define its role in the pathogenesis of IDDM . Peripheral blood mononuclear cells (PBMC) from individuals with newly diagnosed IDDM or at varying levels of risk for the disease were stimulated in vitro with the entire 42-kDa internal domain of IA-2 (amino acids 603-979), a series of control antigens (glutathionine-S-transferase, tetanus toxoid, Candida albicans, mumps, bovine serum albumin), and a mitogen (phytohemagglutinin) . The frequency and mean stimulation index of PBMC proliferation against IA-2 was significantly higher in newly diagnosed IDDM subjects (14 of 33 {42%}; 3.8+/-4.5 at 10 microg/ml) and autoantibody-positive relatives at increased risk for IDDM (6 of 9 {66%}; 3.9+/-3.2) compared with autoantibody-negative relatives (1 of 15 {7%}; 1.8+/-1.0) or healthy control subjects (1 of 12 {8%}; 1.5+/-1.0) . The frequencies of cellular immune reactivities to all other antigens were remarkably similar between each subject group . Sera from 58% of the newly diagnosed IDDM patients tested were IA-2 autoantibody positive . Despite investigations suggesting an inverse association between humoral and cellular immune reactivities against islet-cell-associated autoantigens, no such relationship was observed (rs=0.18, P=0.39) with respect to IA-2 . These studies support the autoantigenic nature of IA-2 in IDDM and suggest the inclusion of cellular immune responses as an adjunct marker for the disease.

J Indian Med Assoc, 1997 Nov, 95(11), 573 - 5
A clinicopathological study of infections in burn patients and importance of biopsy; Bariar LM et al.; Thirty-four patients with burns were subjected to the clinicopathological study with special reference to culture of the wound and histological examination of the burned tissue . The commonest cause of burn was thermal burn (91.18%) followed by electric burn (5.88%) and chemical burn (2.94%) . The maximum number of patients belonged to second and third decades of life (61.78%) . Females were commonest victims, with male:female ratio was 1:1.6 . On the 1st to 3rd postburn day (PBD) most of the wound remained sterile and Strept haemolyticus was first bacteria isolated on 1st PBD . Gram-positive and Gram-negative bacteria, specially pseudomonas, invaded the burn wound as early as 3rd PBD, it was more so with the patients of extensive burn . Among the Gram-positive bacteria Staph aureus was most notorious and invaded burn wound very early . Pseudomonas had maximum growth followed by klebsiella and Esch coli, multidrug resistance was more common with pseudomonas . Among the fungal infections Candida albicans had maximum incidence . Positive blood cultures for bacteria were seen during 2nd, 3rd and 4th postburn weeks . Pseudomonas was the commonest bacteria isolated . Biopsies were done in 17 patients and showed maximum incidence of bacterial infection followed by fungal infection . Patients with burn more than 60% of total body surface area (TBSA) had 100% mortality, while patients with 20-30% of TBSA burn had 20% mortality, the overall mortality was 50% . Biopsies of the burn wound played an important role in the accurate diagnosis and thus helped in starting early specific therapy to prevent death from sepsis.

Eur J Surg, 1998 Mar, 164(3), 217 - 22
Severe sepsis in cardiac surgical patients; Michalopoulos A et al.; OBJECTIVE: To elucidate the incidence, determinants, and consequences of severe sepsis after cardiac surgery . DESIGN: Prospective study . SETTING: Cardiac surgical unit, Greece . SUBJECTS: 2615 adult patients having cardiac operations . MAIN OUTCOME MEASURES: Microbiological evidence of sepsis, mortality, and duration of stay in the intensive care unit (ICU) and hospital . RESULTS: Severe sepsis developed in 41/2615 patients (2%), all during their stay in the ICU: there were 30 men and 11 women, mean (SD) age 65 (10) years . It was most common after combined coronary artery bypass grafting and valve-related operations (7/95, 7%), followed by miscellaneous cardiac operations (7/147, 5%), valve replacement (8/359, 2%), and coronary artery bypass grafting (19/2014, 1%) . When the 41 patients who developed severe sepsis were compared with those who did not (n = 2574) by univariate analysis, there were significant differences in age (p = 0.004); type of operation (p < 0.0001); duration of operation (p < 0.001); bleeding that necessitating either reoperation or significantly more blood transfused (p < 0.0001); and the incidence of low cardiac output syndrome (p = 0.0001) . Of the 41 patients with severe sepsis, 19 (46%) had serious operative complications, 40 (98%) had severe complications in the ICU, and 16 (39%) required reintubation for hypoxaemia . Among the 41 there were 54 bacteraemic episodes of which 37 (69%) were caused by gram positive cocci, 6 (11%) by gram negative bacteria, and 11 (20%) by Candida albicans . Staphylococcus epidermidis was the most common pathogen isolated (n = 26, 48%) . Sepsis associated with bacterial infection usually developed during the first two weeks, and that caused by fungal infection was most common after the twentieth postoperative day . Patients with severe sepsis required longer mechanical ventilation (31 (21) days compared with 0.9 (0.1) days); longer stay in the ICU (40 (25) days) compared with 2 (1) days); longer stay in hospital (48 (27) days compared with 10 (2) days); and significantly more of them died (13 (32%) compared with 41 (2%), p < 0.0001 in each case) . CONCLUSIONS: We concluded that severe sepsis mainly developed in cardiac surgery patients with serious operative and postoperative complications and was associated with a longer stay in both ICU and hospital, and a higher mortality.

FEMS Microbiol Lett, 1998 Apr 1, 161(1), 179 - 85
Cloning and characterization of the phenylalanyl-tRNA synthetase beta subunit gene from Candida albicans; Marcilla A et al.; A Candida albicans expression library was constructed from RNA isolated from regenerating protoplasts . A 1.4-kb cDNA clone was used to isolate a genomic fragment . Sequence analysis revealed an open reading frame of 593 amino acids with an overall identity of 63.6% with the phenylalanyl-tRNA synthetase beta subunit (FRS1) of Saccharomyces cerevisiae . We named it CaFRS1 . It is located in a single copy in chromosome R, SfiI fragment M . Its expression showed a decrease during the cell wall regeneration process in protoplasts of both yeast and mycelial cells of C . albicans, suggesting its requirement thereof in initial steps of the cell wall synthesis.

Nurse Pract, 1998 Mar, 23(3), 44 - 6,49-53
Managing patients with vulvovaginal candidiasis; Elliott KA; Epidemiologic studies have demonstrated a continuing increase in the prevalence of vulvovaginal candidiasis . Although in the past most of these infections were caused by Candida albicans, an increasing percentage are caused by non-albicans Candida species that are less sensitive to the most frequently used antifungal agents . An accurate diagnosis of these infections and the subsequent choice of the most appropriate therapy can only be made after a thorough evaluation of the patient . Successful treatment of vulvovaginal candidiasis is dependent on compliance with therapy; thus, the treatment regimen chosen should fit the patient's daily lifestyle . Newer single-dose regimens offer the option of completing therapy with a single treatment for most patients with uncomplicated vaginal candidiasis . Use of topical agents avoids the potential systemic adverse effects and drug interactions that have been noted with oral antifungals . Patient education and support can also enhance satisfaction with the treatment plan and promote compliance.

Yeast, 1998 Mar 15, 14(4), 335 - 45
Purified arginine permease of Candida albicans is functionally active in a reconstituted system; Mukherjee PK et al.; We have for the first time purified arginine permease from a pathogenic yeast, Candida albicans, to homogeneity by affinity chromatography using L-arginine-linked agarose matrix as affinity column . The purified protein (PP) was of 66 kDa with no subunit structure . Two kinetically distinct binding affinities of PP were evident where high affinity binding (S1) revealed a dependence on acidic pH while pH did not have dramatic effect on low affinity (S2) binding . The specificity of L-arginine binding to PP with regard to other amino acids, structural analogues and inhibitors, was essentially similar to arginine transport observed in the intact cells of C . albicans (Rao et al., 1986) . The purified arginine permease was reconstituted into proteoliposomes and its functionality was tested by imposing a valinomycin-induced membrane potential . All the characteristic features of L-arginine transport displayed by the reconstituted system were similar to those observed in intact cells . Thus homogeneous purified arginine permease was also functionally active.

Antimicrob Agents Chemother, 1998 Apr, 42(4), 907 - 10
Activity of voriconazole, a new triazole, combined with neutrophils or monocytes against Candida albicans: effect of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor; Vora S et al.; The antifungal activity of voriconazole (VCZ) was tested against Candida albicans in the absence or presence of polymorphonuclear neutrophils (PMN) or monocytes . In some experiments, VCZ was compared to fluconazole (FCZ) . On a weight basis, VCZ was 10-fold more efficacious than FCZ against C . albicans Sh27 . Against an FCZ-resistant isolate, VCZ at 1 microg/ml produced the same fungistasis as FCZ at 20 microg/ml . VCZ at 0.1 microg/ml collaborated with PMN for enhanced killing to the same extent as FCZ at 1.0 microg/ml . Granulocyte-colony-stimulating factor (G-CSF) enhanced the candidacidal activity of PMN, and it increased the collaboration of PMN with VCZ for killing . Granulocyte-macrophage (GM)-CSF also significantly enhanced both the killing by PMN and the collaboration of PMN with VCZ for killing . VCZ collaborated with GM-CSF-activated monocytes {corrected} for enhanced killing of C . albicans Sh27, and GM-CSF increased this collaboration . Taken together, these data show that VCZ is more potent than FCZ against C . albicans isolates, alone and in collaboration with PMN or monocytes for enhanced killing . In addition, G-CSF- or GM-CSF-activated PMN and monocytes have enhanced collaboration with VCZ compared to that of unstimulated phagocytes with VCZ.

Antimicrob Agents Chemother, 1998 Apr, 42(4), 903 - 6
Pharmacokinetics of two multiple-dosing regimens of D0870 in human immunodeficiency virus-positive patients: a phase I study; De Wit S et al.; D0870 is a triazole with a broad antifungal spectrum, and it has been shown to have both in vitro and in vivo activities against wild-type and fluconazole-resistant strains of Candida albicans . Twenty-two human immunodeficiency virus (HIV)-positive male subjects were enrolled in an open, nonrandomized trial investigating the pharmacokinetics of two different dosing regimens of D0870 and assessing the safety of multiple oral doses of D0870 in HIV-positive subjects and their ability to tolerate multiple oral doses . Nine subjects received an initial loading dose of 50 mg, followed by four once-daily maintenance doses of 10 mg . A further nine subjects received an initial 200-mg loading dose followed by four daily maintenance doses of 25 mg . All subjects were fasting . A single loading dose of 50 mg of D0870 resulted in a mean maximum concentration in serum (Cmax) of 107 +/- 32 ng/ml . Concentrations in plasma were maintained by the 10-mg once-daily dosing regimen as seen by the similar values of the area under the concentration-time curve from 0 to 24 h following dosing on days 1 and 5 and a mean accumulation ratio close to unity (0.90) . The terminal plasma half-life of D0870 in plasma following dosing on day 5 ranged from 23 to 85 h (mean, 49 h) . A single loading dose of 200 mg of D0870 resulted in a Cmax of 431 +/- 186 ng/ml . Concentrations in plasma were again maintained by the 25-mg daily dosing regimen, with the mean accumulation ratio being close to unity (1.17) . The terminal half-life of D0870 in plasma following dosing on day 5 of phase II of the study ranged from 34 to 137 h (mean, 71 h) . In addition, the concentrations achieved in the plasma of these HIV-positive subjects were similar to the values predicted from simulations based on data derived from normal, healthy subjects . D0870 was well tolerated . No serious adverse events were experienced during the course of the study, and all volunteers completed the trial . A total of 15 adverse events were reported, but none were considered to be related to the administration of D0870 and all had resolved by the end of the trial . No changes in the hematology, clinical chemistry, or urinalysis parameters were considered to be related to dosing with D0870 . No clinically significant changes in the electrocardiogram parameters were noted during the trial . The data generated in this trial support further investigation of these regimens with HIV-positive subjects with fluconazole-susceptible or -resistant oropharyngeal candidosis.

Antimicrob Agents Chemother, 1998 Apr, 42(4), 755 - 61
Role of ABC transporters in aureobasidin A resistance; Ogawa A et al.; Aureobasidin A (AbA) has strong antifungal effects arising from an unusual mechanism . We show that AbA interacts with ATP-binding cassette (ABC) transporters in yeast and mammalian cells . We isolated a gene of Saccharomyces cerevisiae that conferred resistance to AbA when the gene was present in multiple copies . The gene was identical to YOR1/YRS1, which confers resistance to oligomycin, reveromycin, and organic anions, none of which have structures similar to that of AbA . We also isolated an aur3R recessive mutant of S . cerevisiae with increased resistance to AbA . Northern hybridization showed that the aur3R mutant expressed not only YOR1 but also the ABC transporter-encoding gene PDR5 at high levels . Genetic studies showed that the aur3R mutant had a mutation in the PDR1 gene, which encodes a transcriptional regulator of PDR5 and YOR1 . Analysis of a yor1 disruptant of the aur3/pdr1 mutant showed that both the functional YOR1 gene and the mutation in PDR1 were necessary for AbA resistance . These results suggest that YOR1 is more important than PDR5 for AbA resistance . We found in Candida albicans a novel gene whose sequence was similar to the sequence of YOR1 in S . cerevisiae . The amino acid sequence of the C . albicans YOR1 homolog showed no significant similarity to the sequences of CDR1 and CDR2, which are ABC transporters of C . albicans . Furthermore, AbA inhibited the efflux of the anticancer agent vincristine through P glycoproteins in cancer cells with multidrug resistance.

Antimicrob Agents Chemother, 1998 Apr, 42(4), 734 - 8
Thirteen-year evolution of azole resistance in yeast isolates and prevalence of resistant strains carried by cancer patients at a large medical center; Boschman CR et al.; Drug resistance is emerging in many important microbial pathogens, including Candida albicans . We performed fungal susceptibility tests with archived isolates obtained from 1984 through 1993 and fresh clinical isolates obtained from 1994 through 1997 by testing their susceptibilities to fluconazole, ketoconazole, and miconazole and compared the results to the rate of fluconazole use . All isolates recovered prior to 1993 were susceptible to fluconazole . Within 3 years of widespread azole use, we detected resistance to all agents in this class . In order to assess the current prevalence of resistant isolates in our hematologic malignancy and transplant patients, we obtained rectal swabs from hospitalized, non-AIDS, immunocompromised patients between June 1995 and January 1996 . The swabs were inoculated onto sheep's blood agar plates containing 10 microg of vancomycin and 20 microg of gentamicin/ml of agar . One hundred one yeasts were recovered from 97 patients and were tested for their susceptibilities to amphotericin B, fluconazole, flucytosine, ketoconazole, and miconazole . The susceptibility pattern was then compared to those for all clinical isolates obtained throughout the medical center . The antifungal drug histories for each patient were also assessed . The yeasts from this surveillance study were at least as susceptible as the overall hospital strains . There did not appear to be a direct linkage between prior receipt of antifungal agent therapy and carriage of a new, drug-resistant isolate . Increased resistance to newer antifungal agents has occurred at our medical center, but it is not focal to any high-risk patient population that we studied . Monitoring of susceptibility to antifungal agents appears to be necessary for optimizing clinical therapeutic decision making.

Am J Ophthalmol, 1998 Apr, 125(4), 552 - 4
Aerobic and anaerobic microbiology of dacryocystitis; Brook I et al.; PURPOSE: To investigate the aerobic and anaerobic microbiology of dacryocystitis . METHOD: Retrospective review of the 62 clinical and microbiologic records collected between 1980 and 1990 . RESULTS: Aerobic or facultative bacteria were recovered in 32 cases (52%), anaerobic bacteria only in 20 cases (32%), mixed aerobic and anaerobic bacteria in seven cases (11%), and fungi in three cases (5%) . A total of 94 organisms (1.5 per specimen), which included 56 aerobic or facultative anaerobic organisms, 35 anaerobic organisms, and three fungi, were recovered . The predominant aerobic and facultative bacteria were Staphylococcus aureus (15 isolates), Staphylococcus epidermidis (13 isolates), and Pseudomonas species (seven isolates) . The most frequently recovered anaerobes were Peptostreptococcus species (13 isolates), Propionibacterium species (12 isolates), Prevotella species (four isolates), and Fusobacterium species (three isolates) . The predominant fungus was Candida albicans (two isolates) . Polymicrobial infection was present in 28 cases (45%) . CONCLUSION: These data highlight the potential importance of anaerobic bacteria in dacryocystitis.

FEBS Lett, 1998 Mar 27, 425(2), 263 - 5
Molecular diversity of sterol 14alpha-demethylase substrates in plants, fungi and humans; Lamb DC et al.; Metabolism of lanosterol (LAN), 24-methylene-24,25-dihydrolanosterol (24-methyleneDHL), dihydrolanosterol (DHL) and obtusifoliol (OBT) by purified human, plant (Sorghum bicolor) and fungal (Candida albicans) sterol 14alpha-demethylase (CYP51; P450(14DM)) reconstituted with NADPH cytochrome P450 reductases was studied in order to elucidate the substrate specificity and sterol stereo- and regio-structural requirements for optimal CYP51 activity . Both human and C . albicans CYP51 could catalyse 14alpha-demethylation of each substrate with varying levels of activity, but having slightly higher activity for their respective endogenous substrates in vivo, dihydrolanosterol for human CYP51 (Vmax = 0.5 nmol/min/nmol CYP51) and 24-methylene-24,25-dihydrolanosterol for C . albicans CYP51 (Vmax = 0.3 nmol/min/nmol CYP51) . In contrast, S . bicolor CYP51 showed strict substrate specificity and selectivity towards its own endogenous substrate, obtusifoliol (Vmax = 5.5 nmol/min/nmol CYP51) and was inactive towards 14alpha-demethylation of lanosterol, 24-methylene-24,25-dihydrolanosterol and dihydrolanosterol . These findings confirm that the presence of the 4beta-methyl group in the sterol molecule renders the plant CYP51 incapable of 14alpha-demethylation thus revealing the strict active site conservation of plant CYP51 during evolution.

Biospectroscopy, 1998, 4(2), 75 - 91
FTIR microspectroscopic study of cell types and potential confounding variables in screening for cervical malignancies; Wood BR et al.; FTIR microscopy was applied to the analysis of cell types and other variables present in Pap smears to ascertain the limitations of infrared spectroscopy in the diagnosis of cervical cancer and dysplasia . It was found that leukocytes, and in particular lymphocytes, have spectral features in the phosphodiester region (1300-900 cm{-1}) suggestive of what has previously been described as changes indicative of malignancy . Endocervical cells and fibroblasts have similar spectral features to HeLa cells and consequently could also confound diagnosis . The use of ethanol as a fixative and dehydrating agent results in retention of glycogen in cervical cell types and thus minimizes spectral changes in the glycogen region due to sampling technique . Spectra of seminal fluids exhibit strong bands in the phosphodiester/carbohydrate region; however, sperm contamination should be easily detectable by the presence of a distinctive doublet at 981/968 cm(-1) . Erythrocyte spectra exhibit a reduction in glycogen band intensity, but can be discerned by a relatively low-intensity nu(s) PO2- band . Endocervical mucin spectra exhibit a reduction in glycogen bands and a very pronounced nu(s) PO2- band, which is similar in intensity to the corresponding band in HeLa cells . Thrombocytes have strong bands in the phosphodiester region, but thrombocytes can be discerned from other cell types by the presence of two small broad bands at 980 and 935 cm(-1) . Candida albicans is characterized by strong bands in the polysaccharide region which could potentially obscure diagnostic bands if C . albicans is present in large numbers . Spectra of bacteria common to the female genital tract, in general, also have strong absorptions in the polysaccharide region; however, bacterial contamination is usually minimal and would not be expected to obscure cervical cell spectra . Nylon threads and bristles from cervical sampling implements produce characteristic IR profiles which allow for easy identification . Given the number of potential confounding variables associated with cervical cytology, a multivariate statistical or neural network analysis would appear to be necessary before the implementation of FTIR technology in clinical laboratories.

Fortschr Med, 1998 Feb 28, 116(6), 22 - 8
{Candida and the gastrointestinal tract . A medical-research evaluation}; Nolting S et al.; In immunocompetent persons, Candida species are members of the normal flora of the gastrointestinal tract . Budding yeasts, in particular Candida albicans, can, however, in patients with a corresponding disposition, spread topically and systemically, that is, they may become pathogenic . In hematological/oncological patients with severe immunodeficiency, for example, the mycelium may infiltrate the muscularis mucosae, with involvement also of the vascular system . The relationships between recurrent diarrhea and Candida are still discussed controversial; various data do, however, suggest that massive colonization with Candida might well represent a(n additional) diarrhea-provoking factor . Similar considerations may also be assumed to apply to diarrhea induced by antibiotic therapy . For immunocompetent persons, guidelines exist for the yeast cell count in the stools . The interpretation of quantitative findings must, however, always be made on an individual basis and against the background of clinical symptoms and/or any particular predisposition of the patient . Reliable treatment of superficial candidasis can be achieved with oral polyene antifungal antibiotics (nystatin, amphotericin B).

J Bacteriol, 1998 Apr, 180(8), 2079 - 86
Isolation and characterization of EPD1, an essential gene for pseudohyphal growth of a dimorphic yeast, Candida maltosa; Nakazawa T et al.; Additional copies of the centromeric DNA (CEN) region induce pseudohyphal growth in a dimorphic yeast, Candida maltosa (T . Nakazawa, T . Motoyama, H . Horiuchi, A . Ohta, and M . Takagi, J . Bacteriol . 179:5030-5036, 1997) . To understand the mechanism of this transition, we screened the gene library of C . maltosa for sequences which could suppress this morphological change . As a result, we isolated the 5' end of a new gene, EPD1 (for essential for pseudohyphal development), and then cloned the entire gene . The predicted amino acid sequence of Epdlp was highly homologous to those of Ggp1/Gas1/Cwh52p, a glycosylphosphatidylinositol-anchored protein of Saccharomyces cerevisiae, and Phr1p and Phr2p of Candida albicans . The expression of EPD1 was moderately regulated by environmental pH . A homozygous EPD1 null mutant showed some morphological defects and reduction in growth rate and reduced levels of both alkali-soluble and alkali-insoluble beta-glucans . Moreover, the mutant could not undergo the transition from yeast form to pseudohyphal form induced by additional copies of the CEN sequence at pH 4 or by n-hexadecane at pH 4 or pH 7, suggesting that EPD1 is not essential for yeast form growth but is essential for transition to the pseudohyphal form . Overexpression of the amino-terminal part of Epd1p under the control of the GAL promoter suppressed the pseudohyphal development induced by additional copies of the CEN sequence, whereas overexpression of the full-length EPD1 did not . This result and the initial isolation of the 5' end of EPD1 as a suppressor of the pseudohyphal growth induced by the CEN sequence suggest that the amino-terminal part of Epd1p may have a dominant-negative effect on the functions of Epd1p in the pseudohyphal growth induced by the CEN sequence.

Diagn Microbiol Infect Dis, 1998 Feb, 30(2), 121 - 9
National surveillance of nosocomial blood stream infection due to species of Candida other than Candida albicans: frequency of occurrence and antifungal susceptibility in the SCOPE Program . SCOPE Participant Group . Surveillance and Control of Pathogens of Epidemiologic; Pfaller MA et al.; A national surveillance program of nosocomial blood stream infections (BSI) in the USA between April 1995 and June 1996 revealed that Candida was the fourth leading cause of nosocomial BSI, accounting for 8% of all infections . Forty-eight percent of 379 episodes of candidemia were due to species other than Candida albicans . The rank order of non-C . albicans species was C . glabrata (20%) > C . tropicalis (11%) > C . parapsilosis (8%) > C . krusei (5%) > other Candida spp . (4%) . The species distribution varied according to geographic region, with non-C . albicans species predominating in the Northeast (54%) and Southeast (53%) regions, and C . albicans predominating in the Northwest (60%) and Southwest (70%) regions . In vitro susceptibility studies demonstrated that 95% of non-C . albicans isolates were susceptible to 5-fluorocytosine, and 84% and 75% were susceptible to fluconazole and itraconazole, respectively . Geographic variation in susceptibility to itraconazole, but not other agents, was observed . Isolates from the Northwest and Southeast regions were more frequently resistant to itraconazole (29-30%) than those from the Northeast and Southwest regions (17-18%) . Molecular epidemiologic studies revealed possible nosocomial transmission (five medical centers) . Continued surveillance for the presence of non-C . albicans species among hospitalized patients is recommended.

Pharm Dev Technol, 1997 Aug, 2(3), 275 - 84
Hemolytic and antifungal activity of liposome-entrapped amphotericin B prepared by the precipitation method; Kim JC et al.; A new method of preparing liposomes containing amphotericin B (AmB) was developed with the purpose of reducing the toxicity of AmB without causing a loss in its antifungal activity . The procedure involved the precipitation of AmB and egg phosphatidylcholine (PC) in phosphate buffered saline (PBS, pH 7.4) or tris buffered saline (TBS, pH 7.4) by evaporating methanol and chloroform, which had been previously mixed in the buffer solution, at 4 degrees C and 600 mm Hg . The in vitro toxicity of the precipitated liposomes containing 3, 6, 9, 12, and 15 wt% AmB was compared with that of the film-swollen liposomes containing the equivalent contents of the drug . The hemolytic ability of the precipitated liposomes at 37 degrees C was 50.3% at maximum of the film-swollen liposomes at a dose of 30 micrograms AmB/ml, as measured after 17-hr incubation . The significant reduction in the hemolysis effect may in fact be attributed to the reduced rate of drug release from the precipitated liposomes . The precipitated liposomes were multilayered and aggregates of AmB were embedded in the bilayers . These aggregates of AmB would be responsible for an intensive positive peak around 330 nm and reduced toxicity . Despite the decrease in toxicity, the activity of the precipitated liposomes against Candida albicans remained almost equipotent to that of the film-swollen liposomes . Therefore, liposomes prepared by the precipitation method are less toxic but equally as active.

J Immunol, 1998 Jan 1, 160(1), 284 - 92
The role of recombinant murine IL-12 and IFN-gamma in the pathogenesis of a murine systemic Candida albicans infection; Lavigne LM et al.; Studies on murine candidiasis suggest that resistance to disease is linked to a Th1 response and production of IFN-gamma, while failure to elicit protection is associated with a Th2 response and production of IL-4 and IL-10 . Experimental infection of C57BL/6 mice, IL-12 treatment of these mice, or both infection and IL-12 treatment resulted in a characteristic Th1 cytokine mRNA profile as measured by quantitative competitive PCR . Specifically, little or no IL-4 transcripts were detected, while IFN-gamma message was elevated, particularly with IL-12 treatment . Despite its role in driving increased IFN-gamma expression and production, IL-12 treatment, paradoxically, promoted disease progression in our model . Therefore, we examined the effect of IFN-gamma neutralization on IL-12-induced susceptibility to infection . None of the systemically infected mice receiving IL-12 alone survived, while IL-12- and anti-IFN-gamma-treated mice had a 70% survival rate, similar to that after infection alone . These results suggested that IFN-gamma induced by IL-12 treatment contributed to lethality . However, in separate studies, IFN-gamma knockout mice were more susceptible to infection than their wild-type counterparts, suggesting that IFN-gamma is required for resistance . Nonetheless, infected IFN-gamma knockout mice treated with recombinant murine IL-12 exhibited enhanced resistance, suggesting that the toxicities observed with IL-12 are directly attributable to IFN-gamma and that an optimal immune response to Candida infections necessitates a finely tuned balance of IFN-gamma production . Thus, we propose that although IFN-gamma can drive resistance, the overproduction of IFN-gamma during candidiasis, mediated by IL-12 administration, leads to enhanced susceptibility.

Eur J Obstet Gynecol Reprod Biol, 1998 Mar, 77(1), 107 - 9
Fetal Candida sepsis at midgestation: a case report; Engelhart CM et al.; A patient presented with intrauterine fetal death at 21 weeks . A Candida vaginitis was treated at 18 weeks of gestation . Fatal fetal Candida sepsis caused by Candida albicans can occur in the absence of known risk factors such as prolonged rupture of membranes, the presence of an intrauterine contraceptive device or cervical cerclage.

Dent Mater J, 1996 Dec, 15(2), 220 - 5
Disinfection of removable dentures using ozone; Murakami H et al.; Over time, removable dentures tend to become unsanitary and emit unpleasant odors, and oral mucosa sometimes becomes inflamed or denture stomatitis is caused by denture plaque . Recently, various cleaning products designed to keep removable dentures sanitary have appeared on the market . It is known that denture plaque is mainly composed of Candida albicans (C . albicans), and that ozone seems to inhibit these micro-organisms . Accordingly, a denture cleaner using ozone bubbles (ozone concentration of about 10 ppm) was considered as clinically appropriate because of its strong disinfecting and deodorizing power, and high biological safeness . The effectiveness of this cleaner against C . albicans was investigated using . Results showed that C . albicans decreased to about 1/10 after 30 min and to 1/10(3) after 60 min.

Eur J Cell Biol, 1998 Feb, 75(2), 118 - 27
Characterization of monoclonal antibodies against cell wall epitopes of the insect pathogenic fungus, Nomuraea rileyi: differential binding to fungal surfaces and cross-reactivity with host hemocytes and basement membrane components; Pendland JC et al.; Monoclonal antibodies (MAbs) were generated against epitopes on yeast-like hyphal bodies and hyphae of the entomopathogenic hyphomycete, Nomuraea rileyi . Two MAbs (4C10, 2H4) bind to epitopes common to both hyphal bodies and hyphae, whereas MAb 4E9 binds only to hyphal surfaces . 4C10 and 2H4 appear to be directed towards carbohydrate portions of cell surface mannoproteins, as evidenced by similarities in staining patterns between these MAbs and Concanavalin A on Western blots of N . rileyi cell wall extracts . These MAbs cross-react with antigens on blastospore and hyphal surfaces of two other entomopathogenic fungi, Beauveria bassiana and Paecilomyces farinosus in fluorescence microscopy assays, but do not cross-react with a non-entomopathogenic strain of Candida albicans or with Saccharomyces cerevisiae yeasts . MAb 4C10 also cross-reacts with immunocompetent granular hemocytes from Spodoptera exigua (beet armyworm) and Trichoplusia ni (cabbage looper) larvae and with S . exigua plasmatocytes . Electron microscopy revealed that this MAb binds to a component in cytoplasmic granules in the hemocytes, and that surface labeling may be due to the release of this MAb-positive component upon degranulation . MAb 2H4 does not cross-react with granular hemocytes, but does bind to plasmatocytes and hemocytes that tightly adhere to the substrate in monolayer assays . Additionally, MAb 4C10 specifically labels a basement membrane epitope on S . exigua fat body, suggesting that this antibody binds to mannose residues on extracellular matrix glycoproteins . Cross-reactivity of these N . rileyi MAbs with insect hemocyte and tissue components indicates that fungal surface epitopes can mimic host surface molecules, which could explain why N . rileyi hyphal bodies are not recognized by granulocytes and are able to circulate freely in the hemolymph without binding to basement membranes lining the hemocoel.

Kansenshogaku Zasshi, 1998 Feb, 72(2), 105 - 13
{Survey of fungemia cases during the past seventeen years at Teikyo University Hospital}; Kawakami S et al.; Fungi were isolated from 642 cases (3.5%) of 18,403 blood samples at Teikyo University Hospital during the 17-years period between 1979 and 1995 . The number of fungemia cases began to increase around 1985, reached a peak in 1988, and since then, it has been gradually decreasing . The fungal species of isolates were: (1) Candida albicans in 224 cases (34.9%), (2) C . parapsilosis in 149 cases (23.2%), (3) C . tropicalis in 87 cases (13.6%), (4) C . glabrata in 65 cases (10.1%), (5) Hansenula anomala in 58 cases (9.0%), (6) C . guilliermondii in 24 cases (3.7%), (7) C . famata in 14 cases (2.1%), (8) Trichosporon beigelii in 11 cases (1.7%), (9) C . inconspicua in 5 cases (0.8%) and C . lusitaniae in 5 cases (0.8%), and other yeasts in 33 causes (5.1%) . The number of isolates of C . albicans has been decreasing since 1989, concomitant with the clinical introduction of fluconazole in this hospital . However, the number of fluconazole-insusceptible fungi such as non-albicans Candida and Trichosporon spp . has increased . Fungemia cases infected concomitantly or sequentially with two or more different fungal species have been found occasionally since 1983 and have shown a high mortality rate . The spectrum of the causative organisms of fungemia appears to be, at least, partly influenced, by the usage of antifungal agents, particularly fluconazole.

FEMS Immunol Med Microbiol, 1998 Feb, 20(2), 153 - 7
Antifungal antibiotic hamycin increases susceptibility of Candida albicans to phagocytosis by murine macrophages; Dhuley JN; Hamycin is an antifungal antibiotic produced by Streptomyces pimprina Thirum . In the present study, the effect of hamycin on (a) the phagocytosis of Candida albicans by murine peritoneal macrophages and (b) the cell surface hydrophobicity (CSH) of C . albicans was investigated . Addition of hamycin to the culture of macrophages and Candida cells increased the susceptibility of Candida cells to the phagocytosis by macrophages . Pretreatment of Candida cells with hamycin increased their vulnerability to killing by macrophages . Examination of physico-chemical properties of Candida cell surface showed a significant decrease in the CSH . These findings suggest that the binding of hamycin to Candida cells induces biochemical/physico-chemical alterations of the surface, so that it becomes more susceptible to phagocytosis by murine macrophages.

FEMS Immunol Med Microbiol, 1998 Feb, 20(2), 103 - 9
Sequence analysis of a compound coding-region microsatellite in Candida albicans resolves homoplasies and provides a high-resolution tool for genotyping; Metzgar D et al.; Sequence diversity at a coding-region microsatellite locus of two diploid Candida species was surveyed . Twenty-one alleles from fourteen strains of Candida albicans and three alleles from two strains of the closely related Candida dubliniensis were sequenced . Results show independent length variation in two contiguous hexanucleotide repeats, one non-contiguous hexanucleotide repeat, and two non-contiguous trinucleotide repeats within a 120 bp coding region . A neighboring, non-repetitive 120 bp region showed no variation . The information density of sequence polymorphisms in this region provides a powerful tool for genotyping microorganisms in epidemiological studies, yielding detailed resolution of closely related strains, and clearly distinguishing the two species studied here . The individual length-variable repeat regions are very short (2-8 repeats), demonstrating that even very short microsatellites can show high levels of length variability when surrounded by similarly repetitive DNA . Extensive homoplasy was discovered among the C . albicans alleles, with the majority of overall length categories consisting of alleles with more than one sequence . Our results show that microsatellite length alone should not be used to assume either sequence identity or identity by descent . Microsatellite length mutations appear to have generated the high degree of both inter- and intraspecific polymorphism seen at the ERK1 locus, and form an island of variability in an otherwise well-conserved gene.

Pharm Acta Helv, 1998 Jan, 72(5), 271 - 7
Construction of a model of the Candida albicans lanosterol 14-alpha-demethylase active site using the homology modelling technique; Holtje HD et al.; On the basis of all hitherto known P450 X-ray structures and applying standard homology modelling procedures a three-dimensional model of the lanosterol-14 alpha-demethylase active site was constructed . The modelled active site nicely hosts the natural substrate lanosterol and the substrate-enzyme complex displayed stability in a 70 ps molecular dynamics simulation . The importance of Thr 122 of lanosterol 14 alpha-demethylase for hydrogen bond formation with the 3-hydroxyl group of lanosterol was found to be a characteristic feature of the interaction geometry.

J Bacteriol, 1998 Apr, 180(7), 1771 - 6
The TATA-binding protein (TBP) from the human fungal pathogen Candida albicans can complement defects in human and yeast TBPs; Leng P et al.; Candida albicans is the major fungal pathogen in humans, yet little is known about transcriptional regulation in this organism . Therefore, we have isolated, characterized, and expressed the C . albicans TATA-binding protein (TBP) gene (TBP1), because this general transcription initiation factor plays a key role in the activation and regulation of eukaryotic promoters . Southern and Northern blot analyses suggest that a single C . albicans TBP1 locus is expressed at similar levels in the yeast and hyphal forms of this fungus . The TBP1 open reading frame is 716 bp long and encodes a functional TBP of 27 kDa . C . albicans TBP is capable of binding specifically to a TATA box in vitro, substituting for the human TBP to activate basal transcription in vitro, and suppressing the lethal delta spt15 mutation in Saccharomyces cerevisiae . The predicted amino acid sequences of TBPs from C . albicans and other organisms reveal a striking pattern of C-terminal conservation and N-terminal variability: the C-terminal DNA-binding domain displays at least 80% amino acid sequence identity to TBPs from fungi, flies, nematodes, slime molds, plants, and humans . Sequence differences between human and fungal TPBs in the DNA-binding domain may represent potential targets for antifungal therapy.

Scand J Immunol, 1980, 11(3), 321 - 5
Lymphocyte transformation with mitogens and antigens during normal human pregnancy: a longitudinal study; Birkeland SA et al.; In a longitudinal analysis of immunological changes in normal human pregnancy, twenty-two pregnant women were studied, with blood samples taken before pregnancy, during its course, at delivery, and 3-5 months after delivery . The blood samples were frozen successively, using a cryobiological freezing system, and were stored until the whole longitudinal series was obtained . The collected material for a longitudinal series was then thawed and tested in one seance . Lymphocyte transformation tests were performed with stimulation with the mitogens phytohaemagglutinin (PHA), pokeweed mitogen (PWM), concanavalin A (Con A) and the specific antigens tuberculin purified protein derivative (PPD), Candida albicans (CA), Staphylococcus aureus (SA) and streptokinase/streptodornase (SK/SD) . No changes were found in PHA or in PWM responses, whereas there were significant changes in the Con A response, with lowest values in the middle of pregnancy . PPD, CA, SA and SK/SD responses all reached lowest values towards the end of pregnancy, with increases subsequently, after delivery . These findings seem to show that T suppressor function is increased and B-lymphocyte function decreases during the course of pregnancy.

J Infect Dis, 1998 Apr, 177(4), 1057 - 63
Human immunodeficiency virus type 1 gp160 and gp41 binding to Candida albicans selectively enhances candidal virulence in vitro; Gruber A et al.; Previously, it has been shown that human immunodeficiency virus (HIV)-1 envelope proteins gp160 and gp41 bind to Candida albicans . Whether this interaction affects candidal virulence in vitro was investigated . HIV-1 gp160 or gp120 treatment of C . albicans significantly altered neither growth nor phospholipase activity of the fungus . However, treatment of C . albicans with gp160, but not with gp120, led to an elevation of free and cell-bound aspartate proteinase . In addition, culture supernatants obtained from C . albicans treated with gp160 or gp41, but not with gp120, showed a strong increase in proteinase activity . Finally, C . albicans viable yeast cells treated with gp160 or gp41 and serum were phagocytosed by polymorphonuclear leukocytes to a lesser extent than was C . albicans treated with gp120 and serum or serum alone . These findings suggest that the interaction between HIV-1 gp160 and C . albicans may promote the virulence of C . albicans in HIV-1-positive patients.

Allergy, 1998 Feb, 53(2), 173 - 9
Assay for detecting IgE and IgG antibodies against Candida albicans cell-wall mannan; Akiyama K et al.; In the present study, we assayed mannan-specific IgE and IgG antibodies in samples of serum isolated from blood collected from adult patients with bronchial asthma, using a liquid-phase method with a polysaccharide, mannan (Mn), purified from Candida albicans (C . alb), and investigated the relationships of allergenicity among a crude extract of C . alb, purified Mn, and acid protease (AP), The correlations between the titers of anti-Mn A and anti-Mn B IgE and IgG were very strong, and the levels of inhibition of anti-Mn A IgE and IgG reactions by Mn A and Mn B were almost identical . Although no common allergenicity was observed between Mn A and AP because there was no correlation between the titers of anti-Mn A and anti-AP IgE, and no inhibition of the anti-Mn A IgE reaction by AP, both antigens were found to exist in crude C . alb . The level of inhibition of anti-crude C . alb IgG reaction by Mn A or Mn B was about 60% . Approximately 70% inhibition of the anti-Mn A IgE reaction was observed for eight different fungal allergen extracts, but no inhibition was observed for 11 of the other fungal allergen extracts tested . The above results indicate that common antigenicity was observed between Mn A and Mn B in the human IgE and IgG antibody production system, and the cross-allergenicity observed among some fungi was considered to be the result of the common antigenicity of Mn isoforms.

Microbiology, 1998 Mar, 144 ( Pt 3), 689 - 95
Thigmotropism and stretch-activated channels in the pathogenic fungus Candida albicans; Watts HJ et al.; The direction of growth of hyphae of the pathogenic fungus Candida albicans responds thigmotropically to surface contours by following scratches, ridges and grooves and by penetrating pores . Here it is shown that the thigmotropic response to ridges is attenuated by GdCl3 and verapamil {blockers of stretch-activated (SA) ion channels and L-type calcium channels, respectively} . At low concentrations, both compounds reduced the percentage of hyphae reorienting on contact with a ridge without markedly affecting hyphal extension rate, suggesting a possible role for SA or other calcium channels in the transduction of the thigmotropic response . In addition, patch-clamp recordings demonstrated SA channel activity in the plasma membrane of both yeast and hyphal cells of C . albicans . Two distinct SA channels with conductances of 54 pS and 20-25 pS in 200 mM KCl were observed in protoplasts from yeast cells and one channel of 51 pS was found in protoplasts from hyphal cells.

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