Candida albicans

J Infect Dis, 1996 Dec, 174(6), 1369 - 72
Inhibition of Candida albicans growth by calprotectin in the absence of direct contact with the organisms; Sohnle PG et al.; Calprotectin is a calcium- and zinc-binding protein that is present in neutrophil cytoplasm and abscess fluid supernatants . This protein appears to inhibit microbial growth through competition for zinc; however, experiments to show that calprotectin can inhibit growth of microorganisms across filter membranes have yielded conflicting results to date . To prevent recontamination of the filtrate by zinc in this type of experiment, Candida albicans was cultured on filter membranes placed on top of an agarose gel containing calprotectin . In these studies, calprotectin in the gels underneath did suppress growth on top of the filters, an effect reversible by 30 microM ZnSO4 . In other experiments, the protein did not adhere to the organisms and later suppress their growth . These results indicate that calprotectin inhibits C . albicans growth in the absence of direct contact with the organisms; the findings support a zinc-deprivation mechanism of antimicrobial activity for this protein.

Gene, 1996 Nov 21, 180(1-2), 189 - 96
A novel group I intron in Candida dubliniensis is homologous to a Candida albicans intron; Boucher H et al.; In the present study, we determined the sequence of group I self-splicing introns found in the large ribosomal RNA subunit of Candida albicans, Candida stellatoidea and the recently-described species Candida dubliniensis . It was found that both the intron and ribosomal RNA nucleotide sequences are almost perfectly identical between different C . albicans strains as well as between C . albicans and C . stellatoidea strains . Comparisons of ribosomal RNA sequences suggest that local isolates of atypical C . albicans from individuals infected with human immunodeficiency virus can be assigned to the C . dubliniensis species . C . dubliniensis strains also harbor a group I intron in their ribosomal RNA, as observed in about 40% of C . albicans strains and all C . stellatoidea strains . This novel C . dubliniensis group I intron is identical to the C . albicans and C . stellatoidea intron, except for two widely divergent stem-loop regions . Despite these differences, the C . dubliniensis intron possesses self-splicing ability in an in vitro assay . Taken together, these data support the idea that C . albicans and C . stellatoidea should be joined together as variants of the same species while C . dubliniensis is a distinct but closely related microorganism . To our knowledge, the C . albicans and C . dubliniensis introns are the first example of a pair of homologous group I introns differing only by the presence of apparently facultative sequences in some stem-loops suspected to be involved in stabilization of tertiary structure.

Eur J Biochem, 1996 Nov 15, 242(1), 29 - 35
Exchange of regions of the carboxypeptidase Y propeptide . Sequence specificity and function in folding in vivo; Ramos C et al.; The propeptide of carboxypeptidase Y from Saccharomyces cerevisiae is important for folding of the enzyme . Previous work {Ramos, C., Winther, J.R . & Kielland-Brandt, M . C . (1994) J . Biol . Chem . 269, 7006-7012} suggested that the sequences essential for in vivo folding were situated in the COOH-proximal third of the propeptide . Concentrating on this region we have investigated the functionality of propeptide variants . Using a random mutagenesis approach we found that two segments can be defined: one in which there is a fairly high tolerance for substitution with unrelated sequences and another that has a more strict requirement for sequence conservation . Nevertheless, an overall lack of requirement for propeptide sequence conservation was found by substitution of the carboxypeptidase Y propeptide with that of a highly divergent propeptide sequence from an otherwise similar carboxypeptidase from Candida albicans . This propeptide was partially functional when combined with carboxypeptidase Y . Analysis of the biosynthesis of the mutant forms of the zymogen showed that a fraction of the molecules proceeded from the endoplasmic reticulum with fairly rapid kinetics, while the rest was degraded.

Anal Biochem, 1996 Nov 15, 242(2), 165 - 71
Discrimination between acid and alkali-labile phosphorylated residues on Immobilon: phosphorylation studies of nucleoside diphosphate kinase; Biondi RM et al.; We have critically analyzed current methodologies for distinguishing histidine and serine phosphorylated residues in proteins and report a simple technique that assures a reliable discrimination . Electro-transfer of a phosphorylated enzyme to Immobilon membranes and its treatment at pH 1 and 14 in buffers containing 5% methanol allows unambiguous distinction between serine/threonine and histidine phosphorylation (O-phosphomonoesters and phosphoramide, respectively) since under these conditions only one type of residue is dephosphorylated . The addition of 5% methanol to all buffers was indispensable to deplete phosphate from membranes incubated successively under acid and basic conditions . The technique was applied to the study of nucleoside diphosphate kinase (NDP kinase) phosphorylation . In this enzyme, autophosphorylation of active site histidine is an accepted intermediate step in the catalytic phosphate transfer activity of nucleoside diphosphate kinase (NDP kinase) . Nonetheless, a significant degree of autophosphorylation on other residues has been reported by several laboratories, and the hypothesis has been advanced that this nonhistidine phosphorylation may play an important role in NDP kinase cellular function, signaling the suppression of metastasis in the case of human NDP kinase A . Using this improved method, we show that human, Escherichia coli and Candida albicans NDP kinases are only autophosphorylated on histidine residues . In addition, we present evidence that the presence of phosphoserine after strong acid hydrolysis of the histidine autophosphorylated enzyme is in fact a nonenzymatic transphosphorylation from phosphohistidine due to the harsh acid treatment . This methodology was also applied to in vivo phosphorylation studies of C . albicans NDP kinase . We believe that the technique will be generally useful in histidine phosphorylation screenings.

Proc Natl Acad Sci U S A, 1996 Nov 12, 93(23), 13223 - 8
Candida albicans strains heterozygous and homozygous for mutations in mitogen-activated protein kinase signaling components have defects in hyphal development; Kohler JR et al.; The Candida albicans genes, CST20 and HST7, were cloned by their ability to suppress the mating defects of Saccharomyces cerevisiae mutants in the ste20 and ste7 genes, which code for elements of the mating mitogenactivated protein (MAP) kinase pathway . These Candida genes are both structural and functional homologs of the cognate Saccharomyces genes . The pattern of suppression in Saccharomyces is related to their presumptive position in the MAP kinase cascade . Null alleles of these genes were constructed in Candida . The Candida homozygous null mutants are defective in hyphal formation on some media, but are still induced to form hyphae by serum, showing that serum induction of hyphae is independent of the MAP kinase cascade . The Candida heterozygotes CST20/cst20 and HST7/hst7 are also defective in hyphal formation . This lack of dominance of the wild-type allele suggests that gene dosage is important in Candida.

Proc Natl Acad Sci U S A, 1996 Nov 12, 93(23), 13217 - 22
Signal transduction through homologs of the Ste20p and Ste7p protein kinases can trigger hyphal formation in the pathogenic fungus Candida albicans; Leberer E et al.; The CST20 gene of Candida albicans was cloned by functional complementation of a deletion of the STE20 gene in Saccharomyces cerevisiae . CST20 encodes a homolog of the Ste20p/p65PAK family of protein kinases . Colonies of C . albicans cells deleted for CST20 revealed defects in the lateral formation of mycelia on synthetic solid "Spider" media . However, hyphal development was not impaired in some other media . A similar phenotype was caused by deletion of HST7, encoding a functional homolog of the S . cerevisiae Ste7p protein kinase . Overexpression of HST7 partially complemented the deletion of CST20 . Cells deleted for CST20 were less virulent in a mouse model for systemic candidiasis . Our results suggest that more than one signaling pathway can trigger hyphal development in C . albicans, one of which has a protein kinase cascade that is analogous to the mating response pathway in S . cerevisiae and might have become adapted to the control of mycelial formation in asexual C . albicans.

Neuroimmunomodulation, 1996 Nov-Dec, 3(6), 333 - 5
Interaction of cyclophosphamide and ketamine in vivo; Rojavin MA et al.; Cyclophosphamide 100 mg/kg i.p . increased the duration of ketamine-induced anesthesia in BALB/c mice by 39% . However, combined action of these two substances did not change the number of splenocytes, proliferation of T cells, or phagocytic activity of murine peritoneal macrophages against Candida albicans.

J Investig Allergol Clin Immunol, 1996 Nov-Dec, 6(6), 392 - 4
Candidin in immediate hypersensitivity . Comparison of two antigens; Netto CF et al.; One hundred outpatients in a Clinic of Allergy were submitted to intradermic tests with two types of candidins . The main focus of the research was the comparison of two antigens obtained from the same strains of Candida albicans: one a suspension of yeast cells and the other, a polysaccharide . The readings, taken 20 minutes after the intradermic injections, with positive results were considered as hypersensitivity of the immediate type . Positive results were obtained in 74% of the patients with the yeast cell antigen and in 73% with the polysaccharide antigen . This research mainly deals with the advantages that can be obtained by using the polysaccharide antigen in intradermic tests for evaluating hypersensitivity of the immediate type.

Appl Microbiol Biotechnol, 1996 Nov, 46(4), 400 - 4
Evaluation of 2-{N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino}-2-deoxy-D-glucose, a new fluorescent derivative of glucose, for viability assessment of yeast Candida albicans; Yoshioka K et al.; A new fluorescent derivative of D-glucose, 2-{N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino}-2-deoxy-D-glucose (2-NBDG), which had been previously developed for the analysis of glucose uptake activity by living cells, was investigated to evaluate its applicability for assaying the viability of yeast Candida albicans . Lineweaver-Burk plots showed to uptake of 2-NBDG to be competitively inhibited by D-glucose and not by L-glucose, which suggested the involvement of the glucose transporting system of C . albicans in the uptake of 2-NBDG . A good correlation was obtained between the yeast viability, determined by the plate-count method, and the 2-NBDG uptake activity of yeast cells (correlation constant: r = 0.97) . This is expected to lead to the development of a new fluorescent probe for the determination of yeast cell viability.

J Med Vet Mycol, 1996 Nov-Dec, 34(6), 401 - 6
Development of a method to detect secretory mucinolytic activity from Candida albicans; Colina AR et al.; Ultrastructural examinations of sites where Candida albicans invaded the bowel wall after oral intragastric inoculation of infant mice suggested that blastoconidia are capable of progressive extracellular digestion of the intestinal mucus barrier . Microplate assay methods, based on biotin or digoxigenin-labelling systems, were therefore devised for quantitation of protease and glycosidase activities against the glycoprotein mucin . Labelled mucin was adsorbed on microplate wells, incubated with sample to be assayed for enzyme activity, and the remaining labelled mucin was quantitated by spectrophotometry . Proteolytic activity against mucin was demonstrated using concentrated culture filtrate of C . albicans strain LAM-1, grown in yeast nitrogen base medium containing mucin as sole nitrogen source . The activity was inhibited by boiling for 10 min or by incubation with the aspartyl proteinase inhibitor pepstatin A.

J Med Vet Mycol, 1996 Nov-Dec, 34(6), 393 - 400
Molecular cloning of a gene encoding translation initiation factor (TIF) from Candida albicans; Mirbod F et al.; The differential display technique was applied to compare mRNAs from two clinical isolates of Candida albicans with different virulence; high (potent strain, 16240) and low (weak strain, 18084) extracellular phospholipase activities . Complementary DNA fragments corresponding to several apparently differentially expressed mRNAs were recovered and sequenced . A complementary DNA fragment seen distinctly in the potent phospholipase producing strain was highly homologous to the yeast translation initiation factor (TIF) . The selected DNA fragment was then used as a probe to isolate its corresponding complementary DNA clone from a library of C . albicans genomic DNA . The sequence of isolated gene revealed an open reading frame of 1194 nucleotides with the potential to encode a protein of 397 amino acids with a predicted molecular weight of 43 kDa . Over its entire length, the amino acid sequence showed strong homology (78-89%) to Saccharomyces cerevisiae TIF and (63-80%) to mouse eIF-4A proteins . Therefore, our C . albicans gene was identified to be TIF (Ca TIF) . Northern blot analysis in the two strains of C . albicans revealed that Ca TIF expression is 1.5-fold higher in the potent phospholipase producing strain . The restriction endonuclease digestion of genomic DNA from this potent strain revealed at least two hybridized bands in Southern blot analysis, suggesting two or more closely related sequences in the C . albicans genome.

Microbiology, 1996 Nov, 142 ( Pt 11), 3171 - 80
Changes in cell morphology and carnitine acetyltransferase activity in Candida albicans following growth on lipids and serum and after in vivo incubation in mice; Sheridan R et al.; Candida albicans C316, maintained in the yeast form, showed a proliferation of peroxisomes when grown on triolein or serum as sole carbon source but these structures were absent from glucose-grown cells . Peroxisomes were also apparent in C . albicans obtained after injection into mice and recovery from intraperitoneal washings and kidneys; they may therefore be useful markers to assess a potential in vivo response in cells that are growing in vitro . Transcell-wall structures also occurred in C . albicans grown on triolein or serum, and in cells cultured in vivo, but were not seen in cells grown on glucose . These structures consisted of electron-dense fibrillar material penetrating through the cell wall from the plasmalemma side and protruded out to the exterior of the cell . Endoplasmic reticulum, located at the periphery of the cell, was found to be in close proximity with these cell wall structures . Carnitine acetyltransferase (CAT; EC 2.3.1.7), the key enzyme for the translocation of acetyl units between intracellular compartments, was present in low activities in glucose-grown cells; its activity was increased some 100-fold in triolein-grown cells but only 4-fold in serum-grown cells . It was not possible to assess this activity in the in vivo-cultured cells . Two separate CAT proteins, partially purifed from isolated microchondria and peroxisomes, respectively, were identified, with different specificities and kinetic properties.

J Infect, 1996 Nov, 33(3), 221 - 6
Identification of an amphotericin B resistant strain of Candida albicans using a rapid 3H-glucose incorporation microassay; Sweeney JF et al.; Using a 3H-glucose incorporation assay, antifungal sensitivity testing undertaken on an isolate of Candida albicans cultured from the blood of a bone marrow transplant patient documented resistance to amphotericin B but sensitivity to fluconazole and itraconazole . Information obtained from in vitro antifungal sensitivity testing can be used to direct in vivo antifungal therapy . Widespread application of standardized in vitro antifungal sensitivity testing is needed.

Immunopharmacol Immunotoxicol, 1996 Nov, 18(4), 549 - 69
Impairment of non-specific immunity in patients in persistent vegetative state; Munno I et al.; In fourteen patients in persistent vegetative state (PVS) immune responsiveness was investigated . In particular, we studied the relationship between brain lesions following traumatic injury and immune system . In this respect, phagocytosis and killing of Candida albicans by polymorphonuclear cells (PMN) and monocytes were tested . In addition serum levels of Interferon-gamma (IFN-gamma) were evaluated . The patients come out from PVS by 3-4 month were used as control group . Data shown a profound impairement of phagocytosis and killing of monocytes and low serum levels of IFN gamma when compared with normal values . Taken together, these findings suggest that brain lesions, may affect non-specific immune response.

Curr Genet, 1996 Nov, 30(5), 423 - 31
Molecular cloning of a cDNA encoding enolase from the filamentous fungus, Aspergillus oryzae; Machida M et al.; A 1.6-kbp full-length cDNA for the Aspergillus oryzae enolase gene (enoA) was cloned . The sequenced insert contained a continuous open reading frame of 1314 bp encoding a protein of molecular weight 47 405 . Among all enolases sequenced to-date, the deduced amino-acid sequence showed the highest homology (74.9%) with Candida albicans enolase (ENO1) . Strong codon biases and multiple transcription start sites downstream from CT-blocks in the 5'-flanking region suggested strong expression . enoA mRNA was found to occupy approximately 3% (w/w) of total mRNA of A . oryzae by quantitative RT-PCR . This strong transcription was dependent on the carbon source in the medium and correlated with the growth rate of the mycelium.

Arch Microbiol, 1996 Nov, 166(5), 327 - 35
Study of supramolecular structures released from the cell wall of Candida albicans by ethylenediamine treatment; Mormeneo S et al.; Candida albicans cell wall components were analyzed by ethylenediamine (EDA) treatment . Based on their different solubility properties, the cell wall components produced three fractions (A, B, and C) . Fractions B (EDA-soluble, water-insoluble) and C (EDA-insoluble) contained glucan, chitin, and protein in different proportions . After zymolyase (mainly a beta-glucanase complex) or chitinase treatment of fractions B and C, more polysaccharides and proteins were solubilized by a second EDA treatment, suggesting that the solubility of the polymers in EDA depends on the degree of polymer interactions . Western blot analysis using two monoclonal antibodies (1B12 and 4C12) revealed electrophoretic patterns that were similar in mycelial and yeast morphologies, except that in material obtained from mycelial walls, an additional band was detected with MAb 1B12 . Fluorescence microscopy of cell wall fractions treated with FITC-labeled Con-A, Calcofluor white, and FITC-labeled agglutinin showed that glucan and mannoproteins are uniformly distributed in fractions B and C, while chitin is restricted to distinct patches . Transmission electron microscopy demonstrated that fraction C maintained the original shape of the cells, with an irregular thickness generally wider than the walls . When fraction C was treated with chitinase, the morphology was still present and was maintained by an external glucan layer, with an internal expanded fibrillar material covering the entire cellular lumen . Degradation of the glucan skeleton of fraction C with zymolyase resulted in the loss of the morphology.

J Pediatr, 1996 Nov, 129(5), 688 - 94
Candida albicans arthritis one year after successful treatment of fungemia in a healthy infant; Swanson H et al.; Fungal arthritis in pediatric patients is rare and is most often associated with hematogenous spread to the affected joint . It is generally seen concomitant with, or shortly after, fungemia . We report a case of an immunocompetent patient in whom candidal arthritis developed 1 year after initial fungemia . The initial candidiasis was considered to be adequately treated with amphotericin B . The Candida isolates from the neonatal fungemia and subsequent arthritis were the some as identified by electrophoretic karyotype, restriction fragment length polymorphism analysis, and antifungal susceptibility testing . Pediatric candidal fungemia, arthritis, and their treatments are discussed.

Clin Diagn Lab Immunol, 1996 Nov, 3(6), 740 - 5
Production of T-helper cell subsets and cytokines by lymphocytes from patients with chronic mucocutaneous candidiasis; Kobrynski LJ et al.; Chronic mucocutaneous candidiasis (CMC) is a heterogeneous group of disorders characterized by recurrent and persistent superficial candidal infections . Cytokine-induced dysregulation of T-helper cell function has been described in other immune-deficient states but has not been studied in CMC patients . We studied T-helper cell subsets by flow cytometry and cytokine production by stimulated lymphocytes in six CMC patients, two healthy pediatric controls, and five healthy adult controls . Peripheral blood lymphocytes were stimulated in vitro with phytohemagglutinin or Candida albicans extract, and the production of interleukin-2R (IL-2R), IL-4, IL-10, and gamma interferon in the supernatants was measured by enzyme-linked immunosorbent assay . CMC patients had a decrease in the CD29+/CD29+ cell population compared with the numbers in controls (P < 0.02) . The percentage of CD4+/CD45RA+ cells was greater in patients than in controls, but the difference was not significant . There was no difference in the production of IL-10 or gamma interferon by the patient lymphocytes . CMC patients produced more IL-4 than the controls (P < 0.001), whereas the controls tended to produce more IL-2R than the patients (P = 0.19) . These findings support the concept that a decrease in CD4+/CD29+ T-helper inducer cells along with T-helper cell dysregulation may lead to defective memory responses to antigens in CMC patients and a decrease in cell-mediated immunity due to inhibition of TH1 cells by increased levels of IL-4.

Clin Diagn Lab Immunol, 1996 Nov, 3(6), 645 - 50
Evidence for the presence of immunoglobulin E antibodies specific to the cell wall phosphomannoproteins of Candida albicans in patients with allergies; Kanbe T et al.; To determine the major antigenic component of Candida albicans against immunoglobulin E (IgE) antibodies in the sera of patients with allergies who were positive for IgE antibodies to C . albicans crude antigen in a CAP system, phosphomannoproteins (CAMP/A or CAMP/B for serotype A or B strain, respectively) and their acid-stable portions (CAMP-S/A or CAMP-S/B) were isolated from beta-mercaptoethanol (2-ME) extracts of C . albicans cells of serotypes A and B, and IgE antibodies against these components were compared with those against protein complex and enolase (CAE) fractions isolated from C . albicans cells . The dot blot test, which was used to detect IgE antibodies to the C . albicans antigens, showed that IgE antibodies to the 2-ME extract and phosphomannoprotein fractions were present in the sera of 98.0% (2-ME extract), 96.8% (CAMP/A), 93.2% (CAMP-S/A), 97.2% (CAMP/B), and 81.5% (CAMP-S/B) of the patients, whereas IgE antibodies to the protein complex and CAE fractions were found in the sera of 73.6 and 48.8% of the patients, respectively . The extent of IgE binding to the 2-ME extract and phosphomannoproteins was well correlated with the fluorescence intensities estimated with the CAP system . Furthermore, the results obtained from the inhibition experiment with the CAP system indicated that the binding of IgE antibodies to Candida antigens is strongly inhibited by the phosphomannoprotein fraction and is an indication that the serum of the patients contained IgE antibodies specific to the cell wall phosphomannoproteins of C . albicans . Finally, an initial chemical analysis indicated that the epitopes for IgE antibodies on the phosphomannoproteins is a carbohydrate portion, since the ability of CAMP/A to inhibit the binding of IgE antibodies to the homologous CAMP/A was destroyed after oxidation by sodium periodate but not after digestion with proteinase K.

Genetics, 1996 Nov, 144(3), 991 - 1003
Asm-1+, a Neurospora crassa gene related to transcriptional regulators of fungal development; Aramayo R et al.; This report describes the identification, cloning, and molecular analysis of Asm-1+ (Ascospore maturation 1), the Neurospora crassa homologue of the Aspergillus nidulans stuA (stunted A) gene . The Asm-1+ gene is constitutively transcribed and encodes an abundant, nucleus-localized 68.5-kD protein . The protein product of Asm-1+ (ASM-1), contains a potential DNA-binding motif present in related proteins from A . nidulans (StuA), Candida albicans (EFGTF-1), and Saccharomyces cerevisiae (Phd1 and Sok2) . This motif is related to the DNA binding motif of the Swi4/Mbp1/Res family of transcription factors that control the cell cycle . Deletion of Asm-1+ destroys the ability to make protoperithecia (female organs), but does not affect male-specific functions . We propose that the APSES domain (ASM-1, Phd1, StuA, EFGTF-1, and Sok2) defines a group of proteins that constitute a family of related transcription factors involved in the control of fungal development.

Antimicrob Agents Chemother, 1996 Nov, 40(11), 2622 - 5
Effect of fluconazole on viability of Candida albicans over extended periods of time; Sohnle PG et al.; The treatment of chronic mycoses may expose the infecting organisms to antimicrobial agents for extended periods of time . It is possible that an azole antifungal drug such as fluconazole, with primarily fungistatic activity in standard in vitro susceptibility tests, might be able to damage the fungal cells and reduce their viability over prolonged incubations under nonproliferating conditions . To test this possibility, Candida albicans yeast cells were exposed to various concentrations of fluconazole in RPMI 1640 tissue culture medium for 4 h at 37 degrees C, washed free of the drug, and then incubated at 37 degrees C for a 28-day period; enumeration of the remaining CFU at various times during this period revealed no increased loss of viability for the fluconazole-exposed organisms . However, when fluconazole was added to the organisms maintained in distilled water (with or without pretreatment with the drug), a marked reduction of viability was found . At 14 days of incubation with two strains of C . albicans, negative cultures were found for 7 of 10 and 10 of 11 samples, respectively, containing 1.0 microgram of fluconazole per ml versus 0 of 10 and 1 of 11 control samples (P of < 0.01 and 0.001, respectively) . The effect of fluconazole on fungal viability under these conditions became noticeable at approximately 7 days and was greater when the samples were incubated at 37 degrees C rather than 25 degrees C . These findings suggest that fluconazole may have fungicidal effects on fungal cells during prolonged exposures under conditions in which the organisms are prevented from proliferating by lack of nutrients.

Antimicrob Agents Chemother, 1996 Nov, 40(11), 2511 - 6
In vitro interaction between amphotericin B and azoles in Candida albicans; Vazquez JA et al.; The use of azole prophylaxis as a measure to prevent invasive fungal infections in high-risk patients is increasing and is now the standard of care in many institutions . Previous studies disagree on whether preexposure of Candida albicans to azoles affects their subsequent susceptibility to amphotericin B (AmB) . The present in vitro study indicates that azole exposure generates a subpopulation of cells that are not affected by subsequent exposure to AmB . These cells that are phenotypically resistant to AmB tolerated by most cells not exposed to azole . The percentage of cells that convert to phenotypic resistance to AmB varies with the concentration and the azole . Itraconazole is more effective than fluconazole in generating cells that are phenotypically resistant to AmB and that tolerate an otherwise lethal transient exposure to AmB . Until cells that are not exposed to fluconazole are simultaneously challenged with AmB, they are not protected to a significant extent from killing by AmB . Cells that are challenged with continuous exposure to AmB also acquire phenotypic resistance to AmB at increased frequencies by azole preexposure, but this requires that the azole be continuously present during incubation with AmB . In addition, Candida cells taken from mature colonies that are not actively growing are not susceptible to the short-term killing effects of AmB without azole preexposure . The adaptive responses of C . albicans to AmB and potentially other antifungal agents that may result from prior exposure to azoles in vitro or potentially in microenvironments in vivo that induce physiological changes may have major clinical implications.

Artif Organs, 1996 Nov, 20(11), 1191 - 5
Antimicrobial activities of iodinated polystyrene derivatives; Lin KJ et al.; The antimicrobial activities of insoluble halogenated acetamidomethy1-styrene polymers (prepared by covalent bonding of iodine to polystyrene) were assessed as were the factors determining antimicrobial efficacy . The most active materials were selected from chlorinated or iodinated polymers . Antimicrobial activities were assessed for Escherichia coli (ATCC 25922; American Type Culture Collection, Rockville, MD, U.S.A.), Saccharomyces cerevisiae, and Candida albicans by determining time-course changes in microbial counts in vitro . A 2-iodoacetamidomethylstyrene polymer (No.6-I:-CH2I) was found to have the greatest antimicrobial activity against both bacteria and fungi . No.6-I is the first antimicrobial material that did not make an inhibition hollow in the conventional diffusion test or for which conjugated iodine showed antibacterial activity . This material can be introduced into styrene units on the surface of devices by chemical modification . This material was most active at 37 degrees C . For coated dishes, antimicrobial activity depended on the depth or swollen character of the reactive layer . NO.6-I requires not only a minimum width of polymer layer, but also frequent contact with microbes to have an antimicrobial effect . No.6-I is valuable as a new material because it has strong antimicrobial activity by itself but does not release active iodine . This material is expected to have various applications in implantable clinical devices.

South Med J, 1996 Nov, 89(11), 1104 - 7
Infection of a pancreatic pseudocyst due to Candida albicans; Foust RT; A 40-year-old man with diabetes mellitus, congestive heart failure, alcoholic cirrhosis, and chronic pancreatitis had an exacerbation of pancreatitis due to alcohol abuse . His condition deteriorated rapidly with development of apparent sepsis; cultures were negative . He slowly improved with multiple antibiotic therapy and total parenteral nutrition . Serial imaging of the pancreas revealed edematous pancreatitis that evolved initially into a phlegmon and later into multiple pseudocysts . Intermittent fever prompted computed-tomography-directed percutaneous aspiration of the largest pancreatic fluid collection, yielding purulent material that grew only Candida albicans . Subsequently, disseminated candidiasis developed . Despite therapy with amphotericin B and aggressive supportive care, the patient died from multiple organ system failure.

J Am Vet Med Assoc, 1996 Nov 1, 209(9), 1604 - 7
Primary photosensitization related to ingestion of alfalfa silage by cattle; House JK et al.; A herd of 650 Holstein cows was examined for skin disease . Approximately 400 of the lactating adults were affected, but heifers, calves, and nonlactating cows were clinically normal . The condition was characteristic of primary photosensitization . Milk production of the affected cows was normal . Affected cows did not appear to be ill, and none of the cows was icteric . Three of 7 cows had high serum gamma-glutamyltransferase activities, but in the other 4 cows, activity was within the reference range . Serum activities of other hepatic enzymes were within reference ranges in the 7 cows that were examined . Hepatic biopsy specimens from 3 cows were normal . Specimens from 4 other cows had changes that ranged from minimal to mild, chronic, lymphoplasmacytic periportal hepatitis to acute, random, necrotizing hepatitis . Development of photosensitivity was related to ingestion of alfalfa silage . Acetone extracts of the alfalfa silage, but not of other feedstuffs, were found to inhibit growth of Candida albicans under ultraviolet light . Cows experimentally fed a diet composed exclusively of the alfalfa silage developed skin lesions after 6 days, but did not have detectable serum concentrations of phylloerythrin.

J Bacteriol, 1996 Nov, 178(21), 6223 - 6
Isolation and composition of inositolphosphorylceramide-type sphingolipids of hyphal forms of Candida albicans; Wells GB et al.; Hyphal forms of the human pathogen Candida albicans have been found to contain substantial quantities of phosphosphingolipids . These lipids were fractionated into three classes by normal-phase high-performance liquid chromatography . The first class contained equimolar amounts of phosphorus, inositol, phytosphingosines, and fatty acids; their composition and chromatographic behavior suggest that these compounds are inositolphosphorylceramides . The second class contained equimolar amounts of phosphorus, mannosylinositol, phytosphingosines, and fatty acids; their composition and chromatographic behavior indicate that these compounds are mannosylinositolphosphorylceramides . The third class of compounds contained phosphorus, mannosylinositol, inositol, phytosphingosines, and fatty acids in a molar ratio of 2:1:1:1:1; their composition and chromatographic behavior indicate that these compounds are mannosyldiinositolphosphorylceramides . Molecular species in each class differ in the composition of long chain bases and fatty acids; the most abundant long chain bases were C18 and C20 phytosphingosines, and the most abundant fatty acids were hydroxy and nonhydroxy C24-26 . The array of sphingolipids in C . albicans is similar to that of Saccharomyces cerevisiae . Sphingolipids have been shown to be essential in S . cerevisiae, thus these lipids, which are not present in animals, offer a potentially unique target for antifungal chemotherapy against C . albicans.

Infect Immun, 1996 Nov, 64(11), 4714 - 8
Gamma interferon protects endothelial cells from damage by Candida albicans by inhibiting endothelial cell phagocytosis; Fratti RA et al.; Once Candida albicans comes in contact with endothelial cells, it induces cellular injury . This endothelial cell injury may be a mechanism by which blood-borne organisms escape from the intravascular compartment and invade the tissue parenchyma during hematogenous infection . We have been investigating the ability of cytokines to modulate endothelial cell injury caused by C . albicans . Previously we reported that pretreatment of endothelial cells with gamma interferon (IFN-gamma) protects these cells from candidal injury in vitro . In the current study, we examined potential mechanisms of the cytoprotective effects of IFN-gamma . Time course experiments demonstrated that maximal reduction in candidal injury of endothelial cells occurred after the endothelial cells had been exposed to IFN-gamma for at least 72 h . In other studies, we determined that IFN-gamma reduced endothelial cell phagocytosis of C . albicans by 41.3% compared with that of untreated endothelial cells (P < 0.01) . Since endothelial cell phagocytosis of C . albicans is required for damage to occur, inhibition of phagocytosis is likely a mechanism by which IFN-gamma protects endothelial cells from candidal injury . We also found that the cytoprotective effect of IFN-gamma is not mediated by reducing access of the organisms to intracellular endothelial cell iron or by upregulating the synthesis of reactive oxygen intermediates (which could potentially reduce the ability of C . albicans to injure endothelial cells) . Thus, inhibiting endothelial cell phagocytosis of C . albicans may be a mechanism by which IFN-gamma augments the host defense against hematogenously disseminated candidal infections.

Infect Immun, 1996 Nov, 64(11), 4561 - 6
Intravenous injection of Candida-derived mannan results in elevated tumor necrosis factor alpha levels in serum; Garner RE et al.; Intravenous injection of Candida albicans into mice produced elevated serum tumor necrosis factor alpha (TNF-alpha) levels . We hypothesized that immunostimulants released in vivo from C . albicans during fungal sepsis might contribute to the elevated levels of TNF-alpha in serum . We tested this hypothesis in mice with C . albicans mannan (CAM) . Increased serum TNF-alpha levels were observed following intravenous and intraperitoneal injections of CAM . Injection of CAM into mice resulted in increased serum TNF-alpha concentrations that reached 1,200 pg/ml of blood, compared with 2,400 microg/ml of blood following injection of 10 microg of endotoxin . The response to CAM was concentration dependent, requiring a minimum dose of 20 microg of CAM per g of body weight . Sera from mice were tested 30, 60, 90, and 120 min after intravenous injections with CAM . TNF-alpha concentrations were minimal 30 and 120 min after intravenous injection and maximal 60 and 90 min after CAM injection . The relative distribution of CAM in vivo in decreasing order was determined to be as follows: blood > liver > lung > spleen, 90 min following injection of a single 5-mg dose of CAM . CAM was confirmed as the stimulating substance by utilizing anti-CAM antibodies in vivo to block the response . Rabbit anti-mannan antibodies administered by intraperitoneal injection 24 h before CAM injection significantly suppressed (P < 0.05) the accumulation of TNF-alpha in the sera . Dexamethasone administered to mice before intravenous injection of mannan significantly reduced (40 to 90% reduction; P < 0.05) the concentrations of TNF-alpha in the sera of treated mice . Thus, when in vivo CAM clearance mechanisms are exceeded, sufficient CAM may become available to stimulate TNF-alpha production, making CAM an important part of pathogenesis in Candida sepsis.

Infect Immun, 1996 Nov, 64(11), 4514 - 9
Evidence for degradation of gastrointestinal mucin by Candida albicans secretory aspartyl proteinase; Colina AR et al.; A zone of extracellular digestion of the mucin layer around Candida albicans blastoconidia was observed by transmission electron microscopy in the jejunum of mice inoculated intragastrically (G . T . Cole, K . R . Seshan, L . M . Pope, and R . J . Yancey, J . Med . Vet . Mycol . 26:173-185, 1988) . This observation prompted the hypothesis that a putative mucinolytic enzyme(s) may contribute to the virulence of C . albicans by facilitating penetration of the mucus barrier and subsequent adherence to and invasion of epithelial cells . Mucinolytic activity was observed as zones of clearing around colonies of C . albicans LAM-1 grown on agarose containing yeast nitrogen base, glucose, and hog gastric mucin . In addition, concentrated culture filtrate obtained after growth for 24 h in yeast nitrogen base, supplemented with glucose and mucin as the sole nitrogen source, contained proteolytic activity against biotin-labelled mucin which was inhibited by pepstatin A . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the culture filtrate revealed two components of 42 and 45 kDa, with pIs of 4.1 and 5.3, respectively . A zymogram showed that mucin was degraded only by the 42-kDa component, which was also recognized by immunoblotting with an anti-secretory aspartyl proteinase (anti-Sap) 2p monoclonal antibody . The N-terminal sequence of the first 20 amino acids matched that reported for Sap2p . These results demonstrate that Sap2p is responsible for proteolysis of mucin by C . albicans in vitro and may be involved as a virulence factor in the breakdown of mucus and penetration of the mucin barrier by C . albicans.

Proc Natl Acad Sci U S A, 1996 Oct 29, 93(22), 12473 - 7
Molecular markers reveal that population structure of the human pathogen Candida albicans exhibits both clonality and recombination; Graser Y et al.; The life history of Candida albicans presents an enigma: this species is thought to be exclusively asexual, yet strains show extensive phenotypic variation . To address the population genetics of C . albicans, we developed a genetic typing method for codominant single-locus markers by screening randomly amplified DNA for single-strand conformation polymorphisms . DNA fragments amplified by arbitrary primers were initially screened for single-strand conformation polymorphisms and later sequenced using locus-specific primers . A total of 12 single base mutations and insertions were detected from six out of eight PCR fragments . Patterns of sequence-level polymorphism observed for individual strains detected considerable heterozygosity at the DNA sequence level, supporting the view that most C . albicans strains are diploid . Population genetic analyses of 52 natural isolates from Duke University Medical Center provide evidence for both clonality and recombination in C . albicans . Evidence for clonality is supported by the presence of several overrepresented genotypes, as well as by deviation of genotypic frequencies from random (Hardy-Weinberg) expectations . However, tests for nonrandom association of alleles across loci reveal less evidence for linkage disequilibrium than expected for strictly clonal populations . Although C . albicans populations are primarily clonal, evidence for recombination suggests that sexual reproduction or some other form of genetic exchange occurs in this species.

Transplantation, 1996 Oct 27, 62(8), 1182 - 4
Candida albicans osteomyelitis in a liver transplant recipient: a case report and review of the literature; Jonnalagadda S et al.; A 51-year-old man developed fever and back pain 2 months after orthotopic liver transplantation for end-stage liver disease secondary to chronic hepatitis C infection . CT scan demonstrated destructive lesions in T12 suggestive of osteomyelitis . Aspiration biopsy of the vertebra revealed granulomatous inflammation and yeast forms; culture yielded Candida albicans . The patient improved with intravenous amphotericin B and 5-fluorocytosine and did not require surgical intervention . Candida osteomyelitis is a rare condition and to our knowledge it has not been reported before in liver transplant recipients . Awareness of this potential complication may shorten the delay in making the definitive diagnosis, which in turn may increase the likelihood of a response without sequela.

J Med Chem, 1996 Oct 25, 39(22), 4471 - 7
Structure-activity relationships in a series of substituted indolocarbazoles: topoisomerase I and protein kinase C inhibition and antitumoral and antimicrobial properties; Pereira ER et al.; A series of compounds structurally related to staurosporine, rebeccamycin, and corresponding aglycones was synthesized, and their activities toward protein kinase C and topoisomerases I and II were tested together with their in vitro antitumor efficiency against murine B16 melanoma and P388 leukemia cells . Their antimicrobial activities were also examined against a Gram-negative bacterium (Escherichia coli), a yeast (Candida albicans), and three Gram-positive bacteria (Bacillus cereus, Streptomyces chartreusis, and Streptomyces griseus) . To avoid side effects expected with protein kinase C inhibitors, we introduced substitution on the maleimide nitrogen and/or a sugar moiety linked to one of the indole nitrogens to obtain specific inhibitors of topoisomerase I with minimal activities on protein kinase C . As expected, these structures were inefficient on topoisomerase II, and some of them exhibited a strong activity against topoisomerase I . Generally, dechlorinated compounds were found to be more active than chlorinated analogues against both purified topoisomerase I and protein kinase C . On the other hand, opposite results were obtained in the cell antiproliferative assays . These results suggest lack of cell membrane permeability in the absence of the chlorine residue or cleavage of carbon-chlorine bonds inside the cell.

Gene, 1996 Oct 24, 177(1-2), 29 - 34
Candidacidal activity of human salivary histatin recombinant variants produced by site-directed mutagenesis; Driscoll J et al.; Histatin 5 (Hst5) is a 24-amino acid (aa) member of the Hst family that is found in human salivary secretions and exhibits candidacidal activity . Hst5 contains a 13-aa region that alone is capable of killing fungal pathogens and is referred to as the functional domain . To investigate the role of specific aa located within the functional domain, the pRSET bacterial expression system was used to produce recombinant Hst5 (re-Hst5) and several re-variants that were generated by site-directed mutagenesis . The vector pRSETC expresses genes of interest as fusion proteins attached to the carboxy end of an N-terminal His6 tag that binds to nickel (Ni2+) . The re-variants were generated using the polymerase chain reaction (PCR) and had Gly substituted for either the His, Glu or Lys/Arg within the functional domain . PCR products that encoded either the wild-type or variant forms of re-Hst5 were inserted into pRSETC and produced as fusion proteins which were affinity purified from cell lysates by Ni(2+)-Sepharose chromatography . Fusion proteins were digested with CNBr and re-Hsts were purified by reversed-phase high performance liquid chromatography (RP-HPLC) . Re-Hsts were tested in bioassays to measure the ability to kill both Candida albicans (C . albicans) blastoconidia and spheroplasts which were generated by removal of the cell wall . In both assays, re-Hst5 displayed dose-dependent candidacidal activity that was nearly identical to that of native Hst5 purified from human salivary secretions . Re-Hst5 variants with either Glu or Lys/Arg substitutions demonstrated significantly lower candidacidal activity in both assays, while the variant with His mutated showed essentially no activity at physiological concentrations . These results indicate that acidic and basic aa within the functional domain contribute to candidacidal activity and that the His are essential for candidacidal activity . Additionally, since C . albicans spheroplasts were also susceptible to Hsts, the cell wall is not an essential component in the Hst mechanism of candidacidal action.

Biochim Biophys Acta, 1996 Oct 23, 1284(2), 181 - 90
Functional complementation between transmembrane loops of Saccharomyces cerevisiae and Candida albicans plasma membrane H(+)-ATPases; Mason AB et al.; Saccharomyces cerevisiae PMA1 sequences encoding a putative antifungal target site comprising transmembrane loops 1 + 2 and/or 3 + 4 were replaced with the homologous sequences from Candida albicans PMA1 by using PCR-mediated domain transfer . The chimeric pma1 mutants and an isogenic wild type S . cerevisiae strain had similar growth rates, growth yields, glucose-dependent proton pumping rates, acid-activated omeprazole sensitivities, salt tolerances and antifungal sensitivities . The yields and kinetic properties of H(+)-ATPases in plasma membranes of mutant and wild type strains were comparable . Single heterologous transmembrane loops caused deleterious phenotypes at low pH and elevated temperature . Inclusion of both heterologous transmembrane loops fully suppressed the temperature sensitivity caused by heterologous transmembrane loop 1 + 2, partially suppressed the pH sensitivity and gave Candida-like in vitro sensitivity to vanadate, suggesting that the loops operate as a domain . The fully functional chimeric H(+)-ATPase containing C . albicans transmembrane loops 1 + 2 and 3 + 4 demonstrates this domain's complementarity to the equivalent region of the S . cerevisiae enzyme and validates the wild type S . cerevisiae H(+)-ATPase as an antifungal screening target.

Indian J Biochem Biophys, 1996 Oct, 33(5), 420 - 4
Changes in the cellular composition of Candida albicans resistant to miconazole; Sharma S et al.; Biochemical changes that accompany acquisition of miconazole resistance in a single step mutant of C . albicans 3153 were analyzed . Experiments show that resistance to this drug was associated with decrease in total lipids, phospholipids and sterol content . Fluorescence polarization studies with 1,6-diphenyl-1,3,5-hexatriene (DPH) showed decrease in polarization value (p) in resistant cells, thus indicating changes in membrane fluidity . Uptake of {3H} proline by intact cells revealed decrease in K(m) and Vmax of high affinity system (S1) of proline transport in cells resistant to miconazole . Results of this study suggest that membrane sensitivity of miconazole is determined by overall membrane organisation rather than by affinity of antifungal drug(s) for a single membrane component.

J Can Dent Assoc, 1996 Oct, 62(10), 808 - 12
{Denture stomatitis: etiology and clinical considerations}; Girard B Jr et al.; Wearing removable dental prosthesis causes an alteration in the oral microflora . For certain individuals, this new environment is responsible for the development of a particular condition: prosthetic stomatitis . This article reviews the pertinent literature regarding the main predisposing factors causing the disease . It targets the different risk groups and identifies the proposed mechanism for the proliferation of Candida albicans on the palatal side of the prosthesis . Various treatments depending on the severity of the disease are also mentioned.

J Chemother, 1996 Oct, 8(5), 351 - 7
In vitro inhibitory activity of citreoviridin against HIV-1 and an HIV-associated opportunist: Candida albicans; Vieta I et al.; Citreoviridin, a mycotoxin produced by some molds of the genera Penicillium and Aspergillus, inhibits the growth of bacteria of the Bacillus genus . Since significant information was not available on the effects of citreoviridin on eukaryotic cells and viruses, this molecule was tested on CD4+ T-lymphoid cell lines, on HIV-1 and on Candida albicans, which sometimes complicates HIV-infection . Antiviral activity was detected in H9 HTLV IIIB cells, a clone chronically infected by HIV-1 . Citreoviridin reduced p24 in the supernatant of H9 HTLV IIIB in a dose-dependent manner with a pharmacological selectivity index of 2.6 . In C . albicans, the effects of the mold-derivative were evaluated on some parameters associated with pathogenicity and virulence: cellular proliferation, germ tube production, expression of heat shock mannoproteins, release of proteases and phospholipases . At a 12.5 microM dose, citreoviridin showed a marked inhibitory effect on all parameters analyzed . As regards the mechanism of action, it is possible to hypothesize that the effects of citreoviridin may be due to a reduction of protein synthesis, since it inhibited the replication of HIV-1 at post-integrational stages and reduced the intracellular RNA and protein content in C . albicans.

J Clin Pathol, 1996 Oct, 49(10), 861 - 3
Sensitive and universal method for microbial DNA extraction from blood products; Golbang N et al.; A new method of extracting bacterial and yeast DNA from blood products dependent on guanidinium thiocyanate acid extraction and proteinase K treatment is described . In spiked samples the sensitivity per 0.1 ml of serum and blood, respectively, was 26 and 150 colony forming units (cfu) for Escherichia coli, 80 and 120 cfu for Staphylococcus aureus and 20 and 26 cfu for Candida albicans . This compared well with existing methodologies, worked on limited clinical samples and was not pathogen specific.

Immunology, 1996 Oct, 89(2), 295 - 300
The involvement of nitric oxide in the anti-Candida albicans activity of rat neutrophils; Fierro IM et al.; Rat peritoneal neutrophils (PMN) spontaneously release nitric oxide (NO) when incubated in vitro . Addition of the NO synthase inhibitor L-monomethylarginine (L-NMMA) to the PMN reduces NO production and impairs the killing of the yeast Candida albicans, both effects being reversed by L-arginine . These data strongly suggest that oxidative metabolism of L-arginine by PMN is involved in the candidacidal activity of these cells . Rat blood PMN, which do not produce significant amounts of NO, exhibit a reduced killing capacity compared with peritoneal cells, except when they are obtained from lipopolysaccharide (LPS)-treated rats . In this case they produce measurable amounts of nitrite and express high fungicidal activity in vitro . Confirming the candidacidal activity of NO, the exposure of the C . albicans cultures to different concentrations of NO donors leads to a reduction in their survival . The candidacidal activity related to the NO pathway in rat PMN is phagocytosis dependent, since the activity can be inhibited by cytochalasin B . However, the oxidative products of oxygen released by rat PMN do not seem to be involved in their candidacidal activity, as incubation of the cells with phorbol myristate acetate (PMA) increases release of superoxide anion but does not affect the pattern of killing . Our results suggest that NO could be an important candidacidal pathway in rat neutrophils.

J Antimicrob Chemother, 1996 Oct, 38(4), 671 - 7
ZD0870 treatment of murine candidiasis caused by fluconazole resistant isolates of Candida albicans; Najvar LK et al.; Mice were infected intravenously with three fluconazole susceptible and ten fluconazole resistant isolates of Candida albicans then treated with escalating doses of 0.25, 0.5, 1, 2.5, 10 and 40 mg/kg/day of the new antifungal triazole, ZD0870, for 10 days . A minimum protective dose of < 0.25 mg/kg was determined for infections introduced by the three fluconazole susceptible C . albicans and one of the fluconazole resistant isolates whereas doses ranging from 2.5 to 10 mg/kg/day were required for infections induced by seven of the resistant isolates and > or = 40 mg/kg/day for the remainder . Thus, infections caused by fluconazole resistant C . albicans may be successfully treated with ZD0870, though higher doses than those used to treat infections due to susceptible yeast may be required.

J Antimicrob Chemother, 1996 Oct, 38(4), 579 - 87
The effects of antifungal agents on the morphogenetic transformation by Candida albicans in vitro; Hawser S et al.; The effects of antifungal agents, with different mechanisms of action, on the morphogenetic transformation by synchronised yeast-phase Candida albicans cells in vitro and their respective anti-Candida activities are described . MIC data demonstrated that the azoles, amphotericin B and echinocandin were the most active agents against four C . albicans strains . Morphogenetic transformation experiments demonstrated that amphotericin B was significantly better at preventing the transformation, under a variety of test conditions, than the azoles and flucytosine: amphotericin B abolished the transformation at low concentrations while the azoles only prevented the morphogenetic transformation at much higher concentrations (> 100 x MIC).

Infect Immun, 1996 Oct, 64(10), 4406 - 8
Ubiquitin-like epitopes associated with Candida albicans cell surface receptors; Sepulveda P et al.; We have recently reported the cloning of a Candida albicans polyubiquitin gene and the presence of ubiquitin in the cell wall of this fungus . The polyubiquitin cDNA clone was isolated because of its reactivity with antibodies generated against the candidal 37-kDa laminin-binding protein . In the present study, we have further investigated the relationship between ubiquitin and cell wall components displaying receptor-like activities, including the 37-kDa laminin receptor, the 58-kDa fibrinogen-binding mannoprotein, and the candidal C3d receptor . Two-dimensional electrophoretic analysis and immunoblot experiments with antibodies against ubiquitin and the individually purified receptor-like molecules confirmed that these cell surface components are ubiquitinated . In an enzyme-linked immunosorbent assay, polyclonal antisera to each receptor reacted with ubiquitin, thus demonstrating that the purified receptor preparations used as immunogens contained ubiquitin-like epitopes . It is proposed that ubiquitin may play a role in modulating the activity of these receptors and in the interaction of C . albicans cells with host structures.

New Microbiol, 1996 Oct, 19(4), 335 - 43
Morphotypes of Candida albicans and their associations with underlying diseases and source of samples; Riviera L et al.; The morphotyping method was applied to differentiate a series of 276 Candida albicans strains recovered from hospitalized patients, by using a three-digit code based on the characteristics of the fringe outgrowth . By this scheme 32 different morphotypes were identified . An 86% reproducibility was achieved . The aim of this study was to investigate whether significant correlations exist between the morphotypes and: i) some underlying diseases of the patients, ii) the anatomical sources of the samples . The most striking associations were observed between fringeless strains and non-neoplastic diseases of the respiratory tract, and again between continuous filamentous fringe with parallel outgrowth and AIDS . With a significantly high frequency, samples from the genitourinary tract had a very coarse fringe texture.

New Microbiol, 1996 Oct, 19(4), 327 - 34
Observation on the nucleic acids in the chlamydospores of Candida albicans; Vidotto V et al.; The presence of red (RNA) and green (DNA) fluorescent material identifying nucleic acids in the chlamydospores of Candida albicans has been well documented . Red fluorescence in chlamydospores is relatively diffused and ranges from small spots, observed in hyphal cells, to the entire protoplasmic content . Green fluorescence is rarely visible in these structures and, when present it can be observed next to the plasmalemma . The initial percentage values of the two curves related to the cell counts of red fluorescence of the suspensor cells and chlamydospores showed remarkable differences between the two structures . About 54% of the chlamydospores showed red fluorescence compared to about 28% of the suspensor cells . It seems from the results obtained in this study that much RNA was produced and/or accumulated in the chlamydospores and suspensor cells, rather than in mycelium where red fluorescence was not observed . The results obtained sustain the hypothesis that a chlamydospore should he considered a fully functional cell that is morphologically and physiologically active and independent from mycelium.

AIDS, 1996 Oct, 10(12), 1369 - 76
Itraconazole cyclodextrin solution for fluconazole-refractory oropharyngeal candidiasis in AIDS: correlation of clinical response with in vitro susceptibility; Phillips P et al.; OBJECTIVE: To evaluate the efficacy of itraconazole cyclodextrin solution in fluconazole-refractory oropharyngeal candidiasis (OPC), and to correlate clinical outcome with in vitro susceptibility and serum azole levels . DESIGN: A prospective, open-label, intervention study . SETTING: A university hospital, which serves as the provincial HIV referral center . PATIENTS AND INTERVENTIONS: Thirty six HIV-infected individuals referred for fluconazole-refractory OPC were evaluated prospectively between May 1993 and March 1995, including clinical assessment, serum azole levels, and susceptibility testing of Candida spp, isolates . Itraconazole solution was administered orally at a daily dose of 200 mg for 14 days, followed by suppressive therapy . Thirty-four patients were evaluable . MAIN OUTCOME MEASURE: Resolution of oral pseudomembranous lesions . RESULTS: Initial isolates were Candida albicans (n = 33), C . glabrata (n = 1), C . krusei (n = 1), and mixed infection with C . albicans and C . krusei (n = 1) . Fluconazole serum levels obtained at the time of failed therapy ranged from 4.7 to 40 mg/l (median, 12.9 mg/l) . Itraconazole was generally well tolerated . Clinical responses were observed in 65% (22 out of 34) of evaluable cases . Among the responders, relapse had occurred within 2 months for four (36%) out of 11 cases who continued with follow-up . The median fluconazole minimal inhibitory concentration (MIC) was 64 mg/l for isolates from fluconazole-refractory cases, compared with a median of 0.5 mg/l for control isolates (P = 0.002) . The median itraconazole MIC for isolates from fluconazole-refractory cases was 1.25 mg/l, compared with a median of 0.078 mg/l for controls (P = 0.011) . CONCLUSION: A correlation between clinical response and in vitro susceptibility was clearly demonstrated for fluconazole, but not for itraconazole . Itraconazole cyclodextrin solution may be effective for fluconazole-refractory OPC and should be considered prior to salvage therapy with intravenous amphotericin B.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1996 Oct, 82(4), 402 - 7
Clinical characteristics and management responses in 85 HIV-infected patients with oral candidiasis; Silverman S Jr et al.; Eighty-five consecutively seen HIV-positive persons with oral candidiasis were evaluated for clinical characteristics, staging of HIV disease, quantitation of candidal colony formation, and response to systemic antifungal treatment with Nizoral (ketoconazole) . Fifty-five had CD4 counts less than 200 . There was an inconsistent association between clinical signs, patient symptoms, CD4 counts, and candidal colony-forming units . However, there was a trend toward higher colony-forming unit counts (> 500) in patients with lower CD4 cells (< 200) . Sixty-five patients had a complete clinical response to the ketoconazole treatment (200 mg daily for 7 days), even though 81% of posttreatment cultures remained positive . Nonsmokers were more likely to respond to antifungal treatment when compared with smokers, and there was a slight tendency for complete responses when colony-forming unit counts were low . The most common lesion presentation was a combination of the white (pseudomembranous) and red (erythematous) forms . Forty-nine percent had complaints of pain . The variable responses indicated the importance of flexible dose-time and drug considerations in antifungal management . Candida albicans was the predominant species.

Antimicrob Agents Chemother, 1996 Oct, 40(10), 2300 - 5
Susceptibilities of Candida albicans multidrug transporter mutants to various antifungal agents and other metabolic inhibitors; Sanglard D et al.; Some Candida albicans isolates from AIDS patients with oropharyngeal candidiasis are becoming resistant to the azole antifungal agent fluconazole after prolonged treatment with this compound . Most of the C . albicans isolates resistant to fluconazole fail to accumulate this antifungal agent, and this has been considered a cause of resistance . This phenomenon was shown to be linked to an increase in the amounts of mRNA of a C . albicans ABC (ATP-binding cassette) transporter gene called CDR1 and of a gene conferring benomyl resistance (BENr), the product of which belongs to the class of major facilitator multidrug efflux transporters (D . Sanglard, K . Kuchler, F . Ischer, J . L . Pagani, M . Monod, and J . Bille, Antimicrob . Agents Chemother . 39:2378-2386, 1995) . To analyze the roles of these multidrug transporters in the efflux of azole antifungal agents, we constructed C . albicans mutants with single and double deletion mutations of the corresponding genes . The mutants were tested for their susceptibilities to these antifungal agents . Our results indicated that the delta cdr1 C . albicans mutant was hypersusceptible to the azole derivatives fluconazole, itraconazole, and ketoconazole, thus showing that the ABC transporter Cdr1 can use these compounds as substrates . The delta cdr1 mutant was also hypersusceptible to other antifungal agents (terbinafine and amorolfine) and to different metabolic inhibitors (cycloheximide, brefeldin A, and fluphenazine) . The same mutant was slightly more susceptible than the wild type to nocodazole, cerulenin, and crystal violet but not to amphotericin B, nikkomycin Z, flucytosine, or pradimicin . In contrast, the delta ben mutant was rendered more susceptible only to the mutagen 4-nitroquinoline-N-oxide . However, this mutation increased the susceptibilities of the cells to cycloheximide and cerulenin when the mutation was constructed in a delta cdr1 background . The assay used in the present study could be implemented with new antifungal agents and is a powerful tool for assigning these substances as putative substrates of multidrug transporters.

Microbiology, 1996 Oct, 142 ( Pt 10), 2741 - 6
Candida albicans adherence to a human oesophageal cell line; Enache E et al.; The oesophageal epithelium appears to be one of the primary cell targets of Candida albicans in AIDS patients . To study this interaction, we have established an in vitro adherence assay using a human epithelial oesophageal cell line (HET1-A) . When yeast cells were grown in 500 mM D-galactose, adherence increased significantly over cultures prepared in 500 mM D-glucose . In addition to HET1-A cells, adherence of the organism grown in D-galactose to human buccal epithelial cells and a murine alveolar macrophage cell line was also higher . Adherence of yeast cells to HET1-A cells was partially inhibited in the presence of D-glucosamine or N-acetyl-D-glucosamine, but not with D-mannose, D-glucose, L-fucose or D-galactose . Attachment to HET1-A cells was studied using scanning and transmission electron microscopy . Partial phagocytosis of adhering yeast cells was observed occasionally within the first 90 min following infection, as evidenced by the formation of HET1-A pseudopodia in instances of close contact with yeast cells . The influence of D-galactose on cell surface proteins was studied by analysing beta-mercaptoethanol-extracted proteins from yeast cells grown in either 500 mM D-galactose or D-glucose . From D-galactose-grown cells only, a glycoprotein of approximately 190 kDa was observed in Aurodye-stained SDS-PAGE gels and in Western blots using an immunoglobulin fraction (IgG) prepared from sera of rabbits infected with the organism . These studies demonstrate that C . albicans adheres to human oesophageal cells and may utilize cell surface proteins whose synthesis is nutritionally regulated.

J Clin Microbiol, 1996 Oct, 34(10), 2408 - 10
Evaluation of the Microbial Identification System for identification of clinically isolated yeasts; Crist AE Jr et al.; The Microbial Identification System (MIS; Microbial ID, Inc., Newark, Del.) was evaluated for the identification of 550 clinically isolated yeasts . The organisms evaluated were fresh clinical isolates identified by methods routinely used in our laboratory (API 20C and conventional methods) and included Candida albicans (n = 294), C . glabrata (n = 145), C . tropicalis (n = 58), C . parapsilosis (n = 33), and other yeasts (n = 20) . In preparation for fatty acid analysis, yeasts were inoculated onto Sabouraud dextrose agar and incubated at 28 degrees C for 24 h . Yeasts were harvested, saponified, derivatized, and extracted, and fatty acid analysis was performed according to the manufacturer's instructions . Fatty acid profiles were analyzed, and computer identifications were made with the Yeast Clinical Library (database version 3.8) . Of the 550 isolates tested, 374 (68.0%) were correctly identified to the species level, with 87 (15.8%) being incorrectly identified and 89 (16.2%) giving no identification . Repeat testing of isolates giving no identification resulted in an additional 18 isolates being correctly identified . This gave the MIS an overall identification rate of 71.3% . The most frequently misidentified yeast was C . glabrata, which was identified as Saccharomyces cerevisiae 32.4% of the time . On the basis of these results, the MIS, with its current database, does not appear suitable for the routine identification of clinically important yeasts.

J Am Acad Dermatol, 1996 Oct, 35(4), 539 - 42
A U.S . epidemiologic survey of superficial fungal diseases; Kemna ME et al.; BACKGROUND: Large-scale studies performed outside the United States have demonstrated that most cases of onychomycosis and tinea pedis are caused by dermatophytes, primarily Trichophyton rubrum . However, other studies have suggested that yeasts and nondermatophytic molds may play a role, particularly in onychomycosis . OBJECTIVE: This study was undertaken to determine the epidemiology of superficial fungal infections in a U.S . population . METHODS: Fungal cultures were performed on patients with clinically suspected tinea cruris, tinea corporis, tinea capitis, tinea pedis, and onychomycosis . RESULTS: Dermatophytes were the most commonly isolated fungi in each type of superficial fungal disease studied . T . rubrum was the most commonly isolated dermatophyte species, although Trichophyton tonsurans was more common in tinea capitis and equally common in tinea corporis/tinea cruris . In tinea pedis and onychomycosis, dermatophytes appeared in approximately 95% and 82% of isolates, respectively . Candida albicans and nondermatophyte molds played only a minor role in onychomycosis; C . albicans was isolated in 7% of nail cultures and nondermatophytic molds were isolated in 11% . CONCLUSION: These results are in general agreement with other major epidemiologic studies performed outside the United States . Dermatophyte fungi cause most superficial fungal infections.

J Infect Dis, 1996 Oct, 174(4), 888 - 90
Role of extended culture time on synergy of fluconazole and human monocyte-derived macrophages in clearing Candida albicans; Gujral S et al.; Possible enhanced candidacidal synergy of fluconazole and human monocyte-derived macrophages by extended culture time was studied . Synergistic candidacidal activity increased from 1 day to 3 days and was maximal at 6 days . Enhanced synergistic candidacidal activity at 144 h was observed over a 10-fold challenge dose range . Enhanced synergy with culture time was seen with fluconazole-susceptible and -resistant isolates of Candida albicans . These findings emphasize the importance of dosage and an adequate period of therapy.

J Infect Dis, 1996 Oct, 174(4), 821 - 7
Detection and significance of fluconazole resistance in oropharyngeal candidiasis in human immunodeficiency virus-infected patients; Revankar SG et al.; The epidemiology and clinical significance of fluconazole resistance were assessed in a cohort of advanced human immunodeficiency virus (HIV)-infected patients with recurrent oropharyngeal candidiasis . Fifty patients were prospectively evaluated using a novel method of detecting fluconazole resistance with chromogenic media containing fluconazole; results were confirmed with macrobroth testing . Resistant yeasts, defined as MICs > or = 8 micrograms/mL, were detected in 16 (32%) of 50 patients: 7 (14%) had resistant Candida albicans, 7 (14%) had resistant non-C . albicans yeast, and 2 (4%) had mixed resistant yeasts . MICs were > or = 32 in 11 of 16 isolates . Previous fluconazole use and severe immunosuppression were risk factors for resistance . However, 5 of 26 patients had resistant isolates with no prior fluconazole use, and all were severely immunosuppressed . Despite the high prevalence of resistance, 48 patients clinically responded to fluconazole . Fluconazole-resistant C . albicans and non-C . albicans yeast infections are common in patients with advanced immunodeficiency, but clinical efficacy of fluconazole remains high.

Obstet Gynecol, 1996 Oct, 88(4 Pt 2), 704 - 6
Recurrent vulvovaginal candidiasis associated with long-term tamoxifen treatment in postmenopausal women; Sobel JD et al.; BACKGROUND: Symptomatic vulvovaginal candidiasis is rare in postmenopausal subjects because of the estrogen-dependence of this infection . Tamoxifen, a breast-cancer cell estrogen-antagonist, has not previously been reported to predispose to vulvovaginal candidiasis . CASES: Three postmenopausal women, age range 60-81 years (mean 71), were identified with recurrent vulvovaginal candidiasis . In all three cases, new onset of recurrent vulvovaginal candidiasis followed daily tamoxifen therapy . The duration of prior tamoxifen therapy was 1-7 years (mean 3.5) . One patient had diabetes mellitus, an additional risk factor for vulvovaginal candidiasis . In all three patients, Candida glabrata was identified as the causal pathogen, although in two patients symptomatic episodes caused by Candida albicans also occurred . In all cases, diagnosis was easily established using conventional investigations, and eradication of vulvovaginal candidiasis was possible without cessation of tamoxifen . CONCLUSION: Long-term tamoxifen treatment may be complicated by recurrent vulvovaginal candidiasis in postmenopausal women.

J Bacteriol, 1996 Oct, 178(19), 5850 - 2
The mitogen-activated protein kinase homolog HOG1 gene controls glycerol accumulation in the pathogenic fungus Candida albicans; San Jose C et al.; The Candida albicans HOG1 gene (HOG1CA) was cloned by functional complementation of the osmosensitive phenotype associated with Saccharomyces cerevisiae hog1 delta mutants . HOG1CA codes for a 377-amino-acid protein, 78% identical to S . cerevisiae Hog1p . A C . albicans hog1 null mutant was found to be sensitive to osmotic stress and failed to accumulate glycerol on high-osmolarity media.

Arch Microbiol, 1996 Oct, 166(4), 260 - 8
Differential profiles of soluble proteins during the initiation of morphogenesis in Candida albicans; Niimi M et al.; Candida albicans is a dimorphic fungus that can grow either as yeast or as mycelia . The mycelial form may be required for tissue penetration and therefore may have a role in pathogenesis . The protein profiles of the cell-free S100 fraction from budding yeast cells and germ tube-forming cells (an early stage of the transition between yeast and mycelia) were evaluated using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) . Yeast growth or germ tube formation was induced in carbon-starved cells at 37 degrees C by either glucose, galactose or N-acetylglucosamine at pH 4.5 or pH 6.7 . More than 400 constitutively synthesised polypeptides were identified on 2-D PAGE by silver staining . A few polypeptides which seem to reflect the release from carbon starvation were detected, but no polypeptides unique to either morphology were observed . Fractionation of S100 preparations by polyethylenimine or heparin-agarose affinity chromatography, which have been used to detect DNA-binding proteins, revealed several proteins that were synthesised on the resumption of cell growth or in response to pH difference . Heparin-agarose also bound novel polypeptides in the size range 130-200 kDa that were preferentially synthesised in germ tube-forming cells . These results suggest that any protein factors that might exert a regulatory role early in germ tube formation are of low abundance, and that a minor group of soluble proteins involved in C . albicans morphogenesis may be differentially synthesised.

Br Dent J, 1996 Sep 21, 181(6), 209 - 11
Nystatin pastilles and suspension in the treatment of oral candidosis; Millns B et al.; Clinical audit revealed that the treatment of oral candidosis was more successful with nystatin pastilles than with nystatin suspension . The purpose of this investigation was to determine the reasons for this observation . The concentration of nystatin needed to kill 49 consecutive clinical isolates of Candida albicans was measured . The isolates varied in cidal concentrations from 1.875 to 30 U/ml . The time taken to kill these isolates at their cidal concentrations was found to vary from 120 to 300 min . Volunteer studies showed that antifungal activity in the oral cavity was eliminated rapidly after the use of nystatin suspension . In contrast, the polyene could be detected for at least 5 hours after use of the nystatin pastille . The nystatin pastille can be expected to be more effective at killing Candida albicans than the suspension due to its persistent effects.

Fortschr Med, 1996 Sep 20, 114(26), 319 - 21
{Pathogenicity of fungi in the intestines--current status of the discussion}; Scheurlen M; The hypothesis that colonization of the intestinal tract by yeasts (e.g . Candida albicans) can lead to disease in immunocompromised individuals is currently being discussed controversially . Proponents assume that toxins produced by the fungi can trigger such complaints as irritable bowel syndrome of the chronic fatigue syndrome, and that such chronic or recurrent infections may be caused by an intestinal reservoir of yeasts . Opponents of the hypothesis, however, point out that no hard data on the pathogenetic significance of an intestinal reservoir of yeasts are available, controlled studies have failed to demonstrate the effectiveness of antifungal treatment . Discussions are however, hampered by a lack of objective data . The postulated pathomechanisms therefore need to be clarified, diagnostic criteria developed, and the efficacy of the proposed therapeutic measures shown by controlled studies . Until this has been done, assumption about the pathogenicity of yeasts in the bowel, cannot be taken as a basis for binding therapeutic recommendations.

EMBO J, 1996 Sep 16, 15(18), 5060 - 8
Transfer RNA structural change is a key element in the reassignment of the CUG codon in Candida albicans; Santos MA et al.; The human pathogenic yeast Candida albicans and a number of other Candida species translate the standard leucine CUG codon as serine . This is the latest addition to an increasing number of alterations to the standard genetic code which invalidate the theory that the code is frozen and universal . The unexpected finding that some organisms evolved alternative genetic codes raises two important questions: how have these alternative codes evolved and what evolutionary advantages could they create to allow for their selection? To address these questions in the context of serine CUG translation in C.albicans, we have searched for unique structural features in seryl-tRNA(CAG), which translates the leucine CUG codon as serine, and attempted to reconstruct the early stages of this genetic code switch in the closely related yeast species Saccharomyces cerevisiae . We show that a purine at position 33 (G33) in the C.albicans Ser-tRNA(CAG) anticodon loop, which replaces a conserved pyrimidine found in all other tRNAs, is a key structural element in the reassignment of the CUG codon from leucine to serine in that it decreases the decoding efficiency of the tRNA, thereby allowing cells to survive low level serine CUG translation . Expression of this tRNA in S.cerevisiae induces the stress response which allows cells to acquire thermotolerance . We argue that acquisition of thermotolerance may represent a positive selection for this genetic code change by allowing yeasts to adapt to sudden changes in environmental conditions and therefore colonize new ecological niches.

Biochim Biophys Acta, 1996 Sep 13, 1297(1), 1 - 8
D-arabinose dehydrogenase and biosynthesis of erythroascorbic acid in Candida albicans; Kim ST et al.; D-Arabinose dehydrogenase was purified 2750-fold from the cytosolic fraction of Candida albicans to apparent homogeneity, with an overall yield of 3%, by a purification procedure consisting of ammonium sulfate precipitation and DEAE-Sepharose A-50, Sephacryl S-200, Cibacron blue and phenyl-Sepharose CL-4B chromatographies . Gel-filtration chromatography gave an apparent molecular mass of 41 kDa and SDS-PAGE showed only one protein band corresponding to a molecular mass of 42 kDa, indicating that the enzyme is a single polypeptide . The enzyme was optimally active at pH 8.0 and the pI value of the enzyme was 5.0 . The enzyme was relatively stable from pH 4.5 to 7.5 . The optimal temperature for the enzyme activity was 30 degrees C . The activity of the enzyme was inhibited by Hg2+, Fe2+, Zn2+, Cu2+, Mg2+, Mn2+, N-ethylmaleimide and p-chloromercuribenzoic acid . The enzyme catalysed the oxidation of D-arabinose, L-fucose, L-xylose and L-galactose, which have the same configurations of hydroxyl groups at C2- and C3-positions, with apparent K(m) values of 29.2, 28.9, 37.1 and 91.3 mM at pH 8.0, respectively, with 50 microM NADP+ . The enzyme used NADP+ as a coenzyme . Apparent K(m) value at 60 mM D-arabinose for NADP+ was 44.6 microM . NADPH inhibited the enzyme activity competitively with respect to NADP+ (Ki = 78.6 microM) . The amino-terminal sequence of the enzyme was Met-Lys-Leu-Ala-Thr-Glu-Ile-Asp-Phe-X-Leu-Asn-Asn-Gly- . The reaction product was D-arabinono-1,4-lactone, judged from gas-liquid chromatography/mass spectrometry . In C . albicans, D-erythroascorbic acid was formed from D-arabinose by D-arabinose dehydrogenase and D-arabinono-1,4-lactone oxidase.

Mycoses, 1996 Sep-Oct, 39(9-10), 353 - 6
Increased prevalence of oral Candida albicans serotype B in homosexual men: a comparative and longitudinal study in HIV-infected and HIV-negative patients; Torssander J et al.; Several investigators have shown a comparatively high prevalence of Candida albicans serotype B among HIV-infected individuals . We serotyped oral C . albicans strains from 50 HIV-infected homosexual men, 39 HIV-seronegative homosexual men and 40 clinical oral isolates of a control group . The prevalence of serotype B was significantly higher in homosexual men, regardless of HIV serostatus, than in the control subjects . We suggest that the reported high prevalence of serotype B among AIDS patients in Europe and the USA simply reflects the high proportion of homosexual men among HIV-infected patients . In 22 subjects, oral C . albicans isolates were obtained at two or more time points, up to 8 years apart . No change in serotype was observed over time . The serotype prevalences in HIV-infected patients with oral thrush or AIDS-defining illness were similar to the group of homosexual men as a whole, indicating that there is no serotype-related variation in pathogenicity.

Mycoses, 1996 Sep-Oct, 39(9-10), 347 - 51
Fluorometric determination of acid proteinase activity in vulvovaginal candidosis; Kilic N et al.; Vulvovaginal candidosis is one of the most frequent disorders in obstetrics and gynaecology . Candida albicans is commonly considered to be the true vaginopathic agent . The secreted acid proteinase might be especially relevant in the pathogenesis of vulvovaginal candidosis . A fluorometric determination of acid proteinase activity of clinical C . albicans isolates was carried out during the present work using fluorescamine . L-Leucyl-L-alanine was included as an internal standard and the results were expressed as nmoles of leucylalanine equivalents h-1 per 2 x 10(4) cells . The 13 isolates were taken from non-diabetic, non-pregnant women aged 22-35 years with vulvovaginal candidosis . Candida albicans ATCC 44858 was used as a control . The proteinase activity in culture supernatants was detectable starting from the mid- to late- exponential phase of growth, peaked between 30 and 46 h, and then declined . The control had an activity of 2.72 nmol h-1 per 2 x 10(4) cells, whereas eight of the samples had a lower activity (1.05 nmol h-1 per 2 x 10(4) cells on average) and five of the samples had a higher activity (6.53 nmol h-1 per 2 x 10(4) cells on average) . The fluorometric determination of acid proteinase activity was found to be more reproducible and sensitive than the previously used spectrophotometric determinations.

Mycoses, 1996 Sep-Oct, 39(9-10), 341 - 6
Detection of Candida albicans DNA with a yeast-specific primer system by polymerase chain reaction; Wildfeuer A et al.; The in vitro and in vivo selectivity and sensitivity of a yeast-specific primer system was investigated . A two-step polymerase chain reaction (PCR) was used: the first amplified a 245-bp fragment of the gene for cytochrome P450L1A1 and the second a product of 193 bp . This nested PCR produced an approximately 1000-fold increase in the sensitivity of the test for Candida albicans DNA compared with the first primer pair . The lower level of sensitivity of the test in physiological saline and tissue homogenate was about 10 C . albicans cells ml-1 . On the other hand, the sensitivity of the nested PCR method was reduced by a factor of more than 1000 when C . albicans was fixed with 4% formalin . After i.v . injection of different doses of C . albicans into mice, the yeast could be demonstrated in blood and in six different organs . The nested PCR was to some extent more sensitive than culturing for the detection of the yeast in the specimens of organs such as lung, cardiac muscle, liver, kidneys and brain . In contrast, in blood and spleen the culture was superior to the PCR technique used . Nested PCR is thus a useful additional method for the demonstration of yeasts.

Mycoses, 1996 Sep-Oct, 39(9-10), 329 - 39
Clinical use of oral nystatin in the prevention of systemic candidosis in patients at particular risk; Schafer-Korting M et al.; Systemic candidosis is currently a major concern among certain groups of patients at particular risk because of recent treatment modalities . To prevent spread of Candida albicans, in particular, from the orogastrointestinal tract antimycotic treatment would appear beneficial . So far, however, suitable drugs are rare . Polyenes, and in particular oral nystatin, are the main ones considered so far . More recently, the oral azoles have provided therapeutic alternatives . In this review the current role of nystatin and, in particular nystatin tablets, which are better accepted than suspensions at higher dose levels, is described, focusing on efficacy and safety as determined in controlled trials . Recent evidence suggests that oral application of nystatin tablets can be considered both efficacious and safe in the appropriate context . The relative potency of oral nystatin and systemic azoles, particularly ketoconazole and fluconazole, awaits final determination.

Minerva Med, 1996 Sep, 87(9), 433 - 5
{Symptomatic efficacy of clotrimazole combined with surgery in the treatment of recurring anal and genital condylomatosis in seropositive and AIDS patients}; Erenbourg L et al.; During their daily surgical activity in Hiv-positive and AIDS patients the authors have seen in surgical treatment of anogenital recurring acuminate condylomata (treated with diatermocoagulation) in 1995 that prescribing elotrimazole ointment in Candida albicans infection of the same regions there was and amelioration or the healing of the mycotic infection but also a net lowering of the condylomata and persistent latency (disease-free time) till to the next local recurrence . So they have hypothesized and antiviral and antiblastic effect of elotrimazole . Under this heading they have done a literature research discovering that in 1995, only some months ago, two fundamental works have been published on this intriguing topic . They demonstrate at an experimental level in mice a strong antiviral and antiblastic activity of clotrimazole . This new fascinating field is completely open to future research and the consequences at a therapeutic level will be of utmost importance.

APMIS, 1996 Sep, 104(9), 623 - 8
Biotypes of oral Candida albicans isolates in a Tanzanian child population; Matee MI et al.; Although biotypes of Candida albicans from adult populations, especially in the West, have been described, there are no data either from a child population, or from the African continent . Hence a total of 200 oral C . albicans isolates from Tanzanian children aged 6-24 months were biotyped using two commercially available API micromethod kit systems and a boric acid resistance test . The predominant biotypes, which comprised two thirds of the organisms isolated, were J1S (19.5%), A1S (16.0%), J1R (14.5%), A1R (9.5%) and P1R (7.5%) . In total, 16 new biotypes comprising 44 (22%) isolates which have not hitherto been described were found in this Tanzanian population and, of these, the P1R biotype predominated with 15 (7.5%) isolates . There was no significant association between predominant biotypes (with clusters > or = 15 isolates) and age, gender, breast feeding and malnutrition . These data indicate that the biotype profile of C . albicans isolates may differ in paediatric and adult populations, and/or global distribution of various subtypes of this common opportunistic pathogen.

Intern Med, 1996 Sep, 35(9), 707 - 11
Changes in clinical features of fungemia in a Japanese University Hospital over a 12-year period; Mizushima Y et al.; Forty-five patients with fungemia during 1982-1993 (periods I = 1982, II = 1986-1989, III = 1990-1993) in a Japanese university hospital were reviewed to follow changes in the clinical features of fungemia . The percentage of fungi among microorganisms isolated from blood cultures was almost constant (6-10%) throughout the study period . Fungemia was highly associated with use of intravascular catheters, and some changes in clinical features were observed: 1) Candida albicans, C . parapsilosis and C . glabrata were the main isolates, and the number of fungal species showed a tendency to increase . 2) The percentage of patients over 65 years old increased from 36 to 50% . 3) The percentage of patients who were treated with anti-fungal agents and/or removal of catheter increased from 50 to 89 and to 86% . 4) The percentage of patients who died within 28 days after isolation of fungus decreased from 64 to 27% . The improved prognosis was thought to be due to the development of new anti-fungal agents and faster removal of intravascular catheter when infection was suspected.

J Med Vet Mycol, 1996 Sep-Oct, 34(5), 337 - 9
Role of antifungal drug resistance in the pathogenesis of recurrent vulvovaginal candidiasis; Lynch ME et al.; The aetiology of recurrent vulvovaginal candidiasis (RVVC) caused by Candida albicans remains unclear . To adequately address the role of antifungal resistance as a potential mechanism for RVVC, a longitudinal susceptibility analysis of 177 C . albicans isolates collected from 50 C . albicans RVVC patients over a period of 3 months to 7 years was performed . Antifungals tested included clotrimazole, ketoconazole, miconazole, itraconazole and fluconazole . Results showed that all vaginal isolates were uniformly susceptible to all drugs tested and that successive isolates from individual patients did not show increased resistance to any drug despite long-term exposure to azoles . These results suggest that episodes of RVVC caused by C . albicans are rarely of ever attributable to azole antifungal resistance.

J Med Vet Mycol, 1996 Sep-Oct, 34(5), 315 - 22
Cloning and characterization of a cDNA coding for Candida albicans polyubiquitin; Sepulveda P et al.; Immunoscreening of a Candida albicans cDNA library in the expression vector lambda gt11 with rabbit polyclonal antibodies against the 37 kDa cell surface laminin receptor of C albicans resulted in the isolation of a cDNA clone of 0.9 kb . Sequencing of this clone demonstrated a full length open reading frame encoding the polyubiquitin, which contains three tandem copies, head-to-tail spacerless repeats, of the 228 nucleotides coding for the 76 amino acids of the ubiquitin protein, which is identical to that of Saccharomyces cerevisiae . The third copy possesses an extra C-terminal amino acid which is distinct to that found in S . cerevisiae . Northern blot analysis revealed a single mRNA population of about 1 kb present in similar amounts in both yeast and mycelial cells . This indicates that the C . albicans polyubiquitin gene (UBI1) encodes a polyubiquitin precursor protein containing three ubiquitin repeats . Immunofluorescence and Western immunoblotting experiments with polyclonal antibodies against mammalian ubiquitin suggest the presence of ubiquitinated protein moieties in the wall of C . albicans cells.

Microbiologia, 1996 Sep, 12(3), 443 - 8
A Candida albicans gene expressed in Saccharomyces cerevisiae results in a distinct pattern of mRNA processing; Iborra A et al.; Two plasmids (derived from YCplac22 and YEplac112) carrying a Candida albicans gene (including the 5' non-coding promoter sequences) coding for a 30 kDa membrane-bound protein, were used to transform Saccharomyces cerevisiae cells . A 30 kDa protein was immunodetected by Western blot in the membrane fraction of transformants . Northern analysis showed the presence of three mRNA species (of about 1.1, 0.7 and 0.5 kb) hybridizing with the C . albicans gene as a probe . The same result was obtained using the 5' and 3' regions of the gene as probes, whereas only a 1.1 kb mRNA was found in C . albicans and none was detected in S . cerevisiae control transformants . Thus, heterologous expression of this gene in S . cerevisiae results in a distinct pattern of mRNA processing, either due to the location on plasmid vectors and/or to differences in the mRNA processing systems in the two microorganisms.

Comp Immunol Microbiol Infect Dis, 1996 Sep, 19(4), 267 - 74
Effect of beta-endorphin on adherence, chemotaxis and phagocytosis of Candida Albicans by peritoneal macrophages; Ortega E et al.; There is growing evidence about the role of neuroendocrine hormones in the regulation of the immune system . In the present study we have examined effects on different stages of phagocytic function of peritoneal macrophages from BALB/c mice induced by beta-endorphin . Peritoneal macrophages were incubated (30 min at 37 degrees C) in vitro with 0.22, 0.5 or 2200 ng/ml of this hormone . Adherence capacity was evaluated by means of a substrate adherence technique, chemotaxis in Boyden chambers, and ingestion of Candida albicans on migration inhibitory factor (MIF) dishes . No changes in adherence capacity were found . Chemotaxis, however, increased, and concentration of beta-endorphin correlated directly with stimulation . Incubation of macrophages with 0.5 ng/ml of beta-endorphin also stimulated phagocytosis of Candida albicans . These results indicate that beta-endorphin acts on peritoneal marine macrophages, stimulating some stages of their phagocytic function.

J Antimicrob Chemother, 1996 Sep, 38(3), 485 - 97
In-vitro and in-vivo evaluation of a new amphotericin B emulsion-based delivery system; Tabosa Do Egito ES et al.; The in-vitro and in-vivo toxicity and activity of a new emulsion-based delivery system for amphotericin B (AmB-E) and of deoxycholate-amphotericin B (Fungizone) were studied . In vitro, Candida albicans and human red blood cells (RBCs) were treated with either product and dose-response curves for various cellular effects (changes in potassium cell content, haemoglobin leakage from RBCs and colony-forming ability of fungal cells) were obtained . AmB-E was less toxic than Fungizone against human RBCs and equally active against C . albicans cells . In-vivo studies showed that the LD50 of AmB-E and Fungizone in noninfected OF1 mice were 7.24 and 3.46 mg/kg, respectively . The therapeutic efficacy of AmB-E was assessed in murine candidiasis . Firstly, the efficacy of equal doses (0.8 mg/kg) of AmB-E and Fungizone was evaluated in infected mice . Both formulations increased the survival time compared to the control and were equally effective in reducing the cfu counts in the kidney . In the same model of infection, the maximum tolerated doses (MTD) of Fungizone and AmB-E were determined in order to study the efficacies of Fungizone and AmB-E at their respective MTD . AmB-E significantly increased the number of long-term survivors compared with Fungizone (MTD:2 and 1 mg/kg, respectively) . Thus, AmB-E was more effective than Fungizone for treatment of systemic mycoses at the MTD.

J Ethnopharmacol, 1996 Sep, 53(3), 143 - 7
Antimicrobial evaluation of some plants used in Mexican traditional medicine for the treatment of infectious diseases; Navarro V et al.; Twelve methanolic plant extracts from botanical species used in traditional medicine in Morelos, Mexico to cure infectious diseases have been subjected to a screening study to detect potential antimicrobial activity against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans . The antimicrobial activity of the products was evaluated using colonies growing in solid medium, establishing the minimal concentration required to inhibit their in vitro growth (MIC) . The results showed that extracts from Eucalyptus globolus Labill, Punica granatum L., Artemisia mexicana Wild., and Bocconia arborea Watt . possess strong in vitro antimicrobial activity against the tested microorganisms.

Eur J Obstet Gynecol Reprod Biol, 1996 Sep, 68(1-2), 209 - 12
Fetal Candida infection associated with an intrauterine contraceptive device; Marelli G et al.; Fetal Candida infection is rarely described but is often associated with a retained intrauterine contraceptive device (IUCD) . A case of abortion due to Candida infection in a patient wearing an IUCD is reportedPIP: Although fetal intra-amniotic infection with Candida albicans is a rare event, a retained IUD during pregnancy is a major risk factor . Reported in this paper is the case of a 25-year-old woman admitted at 15 weeks' gestation with uterine contractions and vaginal discharge . She reported a history of genital condylomata . Examination revealed premature rupture of the membranes and evidence of an abruptio placentae . An IUD was visible in the uterine cavity . Histologic examination of the placenta revealed growth of Candida pseudohyphae on the surface of the amniotic membranes and at the fetal side of the chorion . A heavy polymorphonuclear infiltrate with areas of necrosis and hemorrhage was noted . Vaginal cultures revealed Candida albicans infection . Most other cases of IUD-associated intra-amniotic candidiasis reported in the literature failed to note whether Candida was present in the vagina or cervix as well . Diagnostic amniocentesis is recommended in all pregnant women with preterm labor and a retained IUD; also indicated is prompt topical antifungal treatment to prevent the spread of Candida to the amniotic cavity .

FEMS Immunol Med Microbiol, 1996 Sep, 15(2-3), 73 - 9
Use of polymorphic short and clustered coding-region microsatellites to distinguish strains of Candida albicans; Field D et al.; We describe the identification of polymorphic microsatellite loci in the pathogenic yeast, Candida albicans . A search for all coding-region microsatellites with more than four repeats that can be found in Candida sequences in GenBank was conducted . Nine such microsatellite sequences consisting of trinucleotide motifs were found . Three of these were perfect microsatellites while the remaining six sequences were found in one imperfect microsatellite and two compound microsatellites . Because of the close proximity of some of these repeats, all could be assayed with six PCR primer pairs . All of these microsatellite sequences were found in five nuclear genes, ZNF1, CCN1, CPH1, EFG1, and MNT2 . Except for a single (CTT)5 serine tract, all coded for polyglutamine tracts . Another locus with seven alleles, a region of the ERK1 protein kinase gene, was also examined, and may be a representative of a new class of highly polymorphic "clustered' microsatellites . Such loci, in which several non-contiguous but closely linked microsatellites are clustered together, may be a useful source of DNA polymorphisms in microorganisms in which long microsatellite sequences are unavailable . All seven regions amplified were polymorphic, having between two and seven variable length alleles in the 11 strains of Candida albicans examined . The results of this and similar searches will facilitate epidemiological and evolutionary studies of Candida and other microorganisms.

Bone Marrow Transplant, 1996 Sep, 18(3), 533 - 40
Bone marrow transplantation for the treatment of haematological disorders in Down's syndrome: toxicity and outcome; Rubin CM et al.; We report 18 patients with Down's syndrome who underwent bone marrow transplantation, and review nine previously published patients . The indications for transplant in the combined group of 27 patients were acute lymphoblastic leukaemia in 14 cases (52%), acute myeloid leukaemia in 11 cases (41%) and aplastic anemia in two cases (7%) . Transplants were autologous in five cases (19%) and allogeneic in 22 cases (81%); of the 22 allogeneic transplants, 16 donors were HLA-matched siblings . In all patients the conditioning regimen included total body irradiation of 7.5 Gy or more, and/or contained cyclophosphamide of 120 mg/kg or more . Seven patients (26%) had fatal pulmonary disease including pneumonitis and pulmonary hemorrhage . Five patients (19%) had significant airway problems including three with severe mucositis who required intubation for airway protection, one with severe mucositis with partial airway obstruction that required observation in the intensive care unit but did not require intubation, and one with Candida albicans laryngitis with development of a glottic web . Nineteen patients (70%) survived beyond 100 days post-transplant . There was no clear association between 100-day survival and the use of any particular agent or regimen used for conditioning or graft-versus-host disease prophylaxis, and the majority of patients tolerated high-dose cyclophosphamide, high-dose cytosine arabinoside, high-dose busulfan, total body irradiation, cyclosporin A, and methotrexate . There appeared to be more early deaths in patients who received the combination of cyclophosphamide and total body irradiation, compared with those receiving the combination of busulfan and cyclophosphamide or those receiving the combination of cytosine arabinoside and total body irradiation . Also, the use of methotrexate was associated with a greater number of early deaths, compared with cyclosporin A . At 3 years, life table estimates of freedom from relapse, relapse-free survival and survival were 75%, 44% and 48%, respectively . The estimated cumulative risk of death due to a non-leukaemic cause at 3 years was 39% . The data show that Down syndrome patients can tolerate the commonly used transplant conditioning regimens with acceptable toxicity; however, there is a strong suggestion in the data that the rates of life-threatening and fatal toxicity are higher than would be expected to occur in patients without Down's syndrome . Patients with Down's syndrome may have a predisposition to fatal pulmonary complications and reversible airway problems during the immediate post-transplant period.

Antimicrob Agents Chemother, 1996 Sep, 40(9), 2221 - 3
Effects of cyclophosphamide and ceftriaxone on gastrointestinal colonization of mice by Candida albicans; Samonis G et al.; Adult male Crl:CD1 (ICR) mice were fed chow containing Candida albicans to induce sustained gastrointestinal colonization by the yeast . Groups of mice were rendered neutropenic with cyclophosphamide and subsequently received ceftriaxone, while other groups received normal saline and served as controls . Stool cultures were obtained immediately before and at the end of treatment . The administration of cyclophosphamide substantially increased the C . albicans counts in the stools of mice . The addition of ceftriaxone to the cyclophosphamide regimen did not significantly increase the level of gastrointestinal colonization by C . albicans . There was no evidence of Candida dissemination to internal organs.

Antimicrob Agents Chemother, 1996 Sep, 40(9), 1998 - 2003
Comparison of a spectrophotometric microdilution method with RPMI-2% glucose with the National Committee for Clinical Laboratory Standards reference macrodilution method M27-P for in vitro susceptibility testing of amphotericin B, flucytosine, and fluconazole against Candida albicans; Rodriguez-Tudela JL et al.; The National Committee for Clinical Laboratory Standards has proposed a reference broth macrodilution method for in vitro antifungal susceptibility testing of yeasts (the M27-P method) . This method is cumbersome and time-consuming and includes MIC endpoint determination by visual and subjective inspection of growth inhibition after 48 h of incubation . An alternative microdilution procedure was compared with the M27-P method for determination of the amphotericin B, flucytosine, and fluconazole susceptibilities of 8 American Type Culture Collection strains (6 of them were quality control or reference strains) and 50 clinical isolates of candida albicans . This microdilution method uses as culture medium RPMI 1640 supplemented with 18 g of glucose per liter (RPMI-2% glucose) . Preparation of drugs, basal medium, and inocula was done by following the recommendations of the National Committee for Clinical Laboratory Standards . The MIC endpoint was calculated objectively from the turbidimetric data read at 24 h . Increased growth of C . albicans in RPMI-2% glucose and its spectrophotometric reading allowed for the rapid (24 h) and objective calculation of MIC endpoints compared with previous microdilution methods with standard RPMI 1640 . Nevertheless, good agreement was shown between the M27-P method and this microdilution test . The MICs obtained for the quality control or reference strains by the microdilution method were in the ranges published for those strains . For clinical isolates, the percentages of agreement were 100% for amphotericin B and fluconazole and 98.1% for flucytosine . These data suggest that this microdilution method may serve as a less subjective and more rapid alternative to the M27-P method for antifungal susceptibility testing of yeasts.

Arzneimittelforschung, 1996 Sep, 46(9), 934 - 6
In vitro synergistic activity of ketoconazole with valproic acid against Candida species; Krajewska-Kulak E et al.; The susceptibility of 148 strains of Candida albicans and 20 strains of Candida species from patients was tested against ketoconazole (CAS 65277-42-1, KTZ), valproic acid (CAS 99-66-1, VPA) and the combination of KTZ and VPA, using Sabouraud's and YNB (yeast nitrogen base) media . Minimal inhibitory concentrations (MIC) with regard of C . albicans determined on diagnostic plates which contained Sabouraud's medium gave values of 49.84 +/- 5.83 mg/l (KTZ) and 202.97 +/- 17.70 mg/l (VPA), and on plates which contained YNB agar, values of 8.06 +/- 0.99 mg/l (KTZ) and 122.57 +/- 12.08 mg/l (VPA) . The combination of KTZ and VPA in various ratios (1:1, 1:2, 2:1) was found to exert synergistic effects against C . albicans on Sabouraud's medium and the mean values of the combinations were: 4.29 +/- 0.37, 5.29 +/- 0.37, 5.02 +/- 0.38 mg/l . These results were significantly different (p < 0.001) compared with KTZ . The findings indicate that VPA, an antiepileptic drug, increases the antifungal activity of KTZ against C . albicans and Candida species in vitro.

Eur J Pediatr, 1996 Sep, 155(9), 756 - 8
Tribological and mycological consequences of the use of a miconazole nitrate-containing paste for the prevention of diaper dermatitis: an open pilot study; Pierard-Franchimont C et al.; The diaper environment increases the coefficient of skin friction and compromises the function of the stratum corneum . In this study, the tribological and mycological benefit of the use of a miconazole nitrate-containing paste on diapered skin was evaluated . A total of 135 instrumental measurements of both erythema and stratum corneum alterations were made on alternate days for 3 weeks in 15 infants . Biometrological parameters were the chromacity a* of the skin and the squamometry index . Cultures testing for Candida albicans were also performed . In the days following the use of the paste, the chromacity a*, the squamometry index and the number of positive cultures of C . albicans were significantly reduced compared to the same evaluations made off treatment . Conclusion: Miconazole nitrate-containing paste reduces the tribological interference between cloth diapers and skin as well as providing the diapered skin with an improved microbial environment.

J Clin Microbiol, 1996 Sep, 34(9), 2246 - 54
Frequency, intensity, species, and strains of oral Candida vary as a function of host age; Kleinegger CL et al.; While the age of the host has been suggested as a determining factor in yeast carriage, no studies in which the genetic relatedness of isolates has been assessed in combination with the frequency and intensity of carriage as a function of host age have been performed in a single geographical locale and over a short time window . Therefore, by using a simple plating protocol to determine the frequency and intensity of carriage, sugar assimilation patterns to determine species, and Southern blot hybridization with the DNA fingerprinting probe Ca3 combined with computer-assisted analysis to determine the genetic relatedness of strains of Candida albicans, yeast carriage was analyzed as a function of age . All test individuals lived in the Iowa City, Iowa, locale and, except for some of the 0.5- to 1.5-year-olds, were dentate . The results demonstrate that for this test population, the frequency, average intensity, predominant species, and genetic relatedness of C . albicans strains varied as a function of host age . In addition, comparison with oral commensal organisms from the Ann Arbor, Mich., locale confirms the geographical specificity of C . albicans strains and the existence of an Iowa City-enriched strain which is most prevalent in elderly individuals.

J Clin Microbiol, 1996 Sep, 34(9), 2106 - 12
Candida albicans serotype analysis by flow cytometry; Mercure S et al.; Candida albicans strains can be assigned to either of two major serogroups, A or B . Antigenic surface determinants present only in serotype A strains allow such a distinction, which has epidemiologic relevance . Reports have established that the relative distributions of the two serotypes can vary depending on the geographic origin of the isolates . A prevalence of susceptibility to an antifungal agent, flucytosine, was also observed with isolates of serotype A . More recently, it was suggested that the occurrence of serotype B isolates in various clinical forms of candidiasis is increasing . However, this latest finding remains controversial since serotyping results vary widely from one laboratory to another because of the lack of standardized methodologies . Difficulty in interpretation of results, which may lead to erroneous serotype identification, is the major setback associated with current methods . For this study, we thus devised a procedure that relies on flow cytometry and that may eliminate ambiguities in serotype determination . The validation of results was achieved with two types of serotype A-specific antisera, Iatron Factor 6 antiserum and an anti-C . albicans antiserum adsorbed on serotype B yeast cells . Agreement between results obtained with these two reagents was 100% with a wide array of Candida strains . These results confirmed the potential of the flow cytometric procedure as a reliable and reproducible method to establish the serotypes of C . albicans strains . Furthermore, some applications of this procedure to the epidemiological study of this human pathogen are presented.

Phytochemistry, 1996 Sep, 43(2), 513 - 20
Xanthones from Hypericum roeperanum; Rath G et al.; Four new xanthones have been isolated from the roots of Hypericum roeperanum . Their structures have been established by a combination of spectroscopic and chemical methods as 1,6-dihydroxy-5-methoxy-4',5'-dihydro-4',4',5'-trimethylfurano- (2',3':3,4)-xanthone (5-O-methyl-2-deprenylrheediaxanthone B), 1,6-dihydroxy-5-methoxy-6',6'-dimethylpyrano-(2',3':3,4)-xanthone (5-O-methylisojacareubin), 1,3,5,6-tetrahydroxy-2-(2',2'-dimethyl-4'-isopropenyl)-cyclopen tanylxanthone (5-O-demethylpaxanthonin) and 1,3,5,6-tetrahydroxy-4-trans-sesquilavandulylxanthone (roeperanone) . In addition, 2-hydroxyxanthone, 5-hydroxy-2-methoxyxanthone, 1,5-dihydroxy-2-methoxyxanthone, 2-deprenyl rheediaxanthone B, isojacareubin and calycinoxanthone D have been isolated and characterized . Some of the isolated xanthones exhibited antifungal activity against Candida albicans.

Lett Appl Microbiol, 1996 Sep, 23(3), 159 - 62
Potential chitinase activating factor from yeast cells of Candida albicans; Jackson DJ et al.; Microsomal chitinase from yeast and hyphal cells of Candida albicans was activated endogenously by incubation at 30 degrees C and exogenously by trypsin . The putative activating factor of yeast cells was separated from chitinase activity by fractionation of lysed protoplasts on an Iodixanol density gradient . The vacuole fraction contained no significant chitinase activity, but was enriched in chitinase activating factor . Activity of microsomal chitinase increased upon incubation with this, but no other gradient factor . Results suggest that the regulatory system governing microsomal chitinase activity, like that governing chitin synthase, involves a 'vacuolar' activating factor in Candida albicans.

Microbiology, 1996 Sep, 142 ( Pt 9), 2515 - 23
Isolation of the mRNA-capping enzyme and ferric-reductase-related genes from Candida albicans; Yamada-Okabe T et al.; The mRNA-capping enzyme (mRNA 5'-guanylyltransferase) gene was cloned from a Candida albicans genomic DNA library by functional complementation of a Saccharomyces cerevisiae ceg1 delta null mutation . This gene, referred to as CGT1 (C . albicans guanylyltransferase 1), can encode a 52 kDa protein that is highly homologous to S . cerevisiae Ceg1p . CGT1 in a single-copy plasmid complemented the lethality of the S . cerevisiae ceg1 delta null mutation and, like S . cerevisiae Ceg1p, bacterially expressed Cgt1p was able to form a stable complex with the GMP moiety of GTP and to synthesize the cap structure in vitro, demonstrating that CGT1 is the C . albicans mRNA 5'-guanylyltransferase gene . CGT1 seemed to exist as a single copy in the C . albicans genome and was actively transcribed into mRNA . Another ORF was found in an opposite strand very close to the CGT1 locus . This gene shared significant sequence homology with S . cerevisiae FRE1, the gene encoding ferric reductase, and therefore was designated CFL1 (C . albicans ferric-reductase-like gene 1) . Despite its sequence homology with S . cerevisiae FRE1, CFL1 mRNA was not induced by iron deprivation, and CFL1 did not complement the slow growth of a S . cerevisiae fre1 delta null mutant in the absence of iron, suggesting that CFL1 is functionally distinct from S . cerevisiae FRE1.

Microbiology, 1996 Sep, 142 ( Pt 9), 2509 - 14
Requirement for the Candida albicans FAS2 gene for infection in a rat model of oropharyngeal candidiasis; Zhao XJ et al.; The virulence of Candida albicans strains deficient in fatty acid synthase activity by virtue of disruption/deletion of the FAS2 gene was examined in a rat model of oropharyngeal candidiasis . The FAS2 alleles of C . albicans CAI4 (delta ura3::imm434/delta ura3::imm434) were sequentially disrupted with a cassette that included a portion of FAS2 from which a 984 bp fragment containing the FAS condensing reaction domain was deleted and replaced with hisG-URA3-hisG sequences . Verification of fatty acid synthase inactivation was obtained from assays of enzyme activity . Strains in which a single allele was disrupted (CFD1 and CFD3) exhibited an approximately 20% reduction in activity, when compared to wild-type . In addition, fatty acid synthase activity was abolished in a FAS2 null mutant strain (CFD2), and growth of CFD2 occurred only when the growth medium was supplemented with Tween 40 and certain fatty acids . Strain CFD2 was avirulent in the rat model, indicating that fatty acid synthase activity is required for C . albicans oropharyngeal infection . Strains with a single FAS2 allele disruption colonized the oral cavity, but the number of cells recovered from infected animals was approximately fivefold less than for the parental strain . The results suggest that FAS may be exploited as a possible target for the development of new antifungal agents.

Biochem J, 1996 Sep 1, 318 ( Pt 2), 437 - 42
In vivo and in vitro folding of a recombinant metalloenzyme, phosphomannose isomerase; Proudfoot AE et al.; Phosphomannose isomerase (PMI) catalyses the interconversion of mannose 6-phosphate and fructose 6-phosphate in prokaryotic and eukaryotic cells . The enzyme is a metalloenzyme which contains 1 mol of zinc per mol of enzyme . Heterologous expression of the cDNA coding for the Candida albicans enzyme in the prokaryotic host Escherichia coli results in an expression level of up to 30% of total E . coli protein . Ten percent of recombinant PMI is expressed in the soluble fraction and 90% in inclusion bodies . Inclusion of a high level of zinc in the fermentation medium resulted in a fourfold increase in soluble protein . Co-expression of the bacterial chaperones, GroES and GroEL, resulted in a proportional twofold increase in soluble PMI while causing an overall decrease in the PMI expression level . Folding denatured PMI in vitro required reductant and zinc ions . The yield of renatured protein was increased by folding in the presence of GroEL and DnaK in an ATP-independent manner . The refolding yield of denatured soluble enzyme from a guanidine solution was threefold higher than that of folding monomerized inclusion body protein solubilized in guanidine hydrochloride . This suggests that a proportion of recombinant protein expressed in E.coli inclusion bodies may be irreversibly denatured.

J Bacteriol, 1996 Sep, 178(18), 5353 - 60
The Candida albicans HYR1 gene, which is activated in response to hyphal development, belongs to a gene family encoding yeast cell wall proteins; Bailey DA et al.; A hyphally regulated gene (HYR1) from the dimorphic human pathogenic fungus Candida albicans was isolated and characterized . Northern (RNA) analyses showed that the HYR1 mRNA was induced specifically in response to hyphal development when morphogenesis was stimulated by serum addition and temperature elevation, increases in both culture pH and temperature, or N-acetylglucosamine addition . The HYR1 gene sequence revealed a 937-codon open reading frame capable of encoding a protein with an N-terminal signal sequence, a C-terminal glycosylphosphatidylinositol-anchoring domain, 17 potential N glycosylation sites, and a large domain rich in serine and threonine (51% of 230 residues) . These features are observed in many yeast cell wall proteins, but no homologs are present in the databases . In addition, Hyr1p contained a second domain rich in glycine, serine, and asparagine (79% of 239 residues) . The HYR1 locus in C . albicans CAI4 was disrupted by "Ura-blasting," but the resulting homozygous delta hyr1/delta hyr1 null mutant displayed no obvious morphological phenotype . The growth rates for yeast cells and hyphae and the kinetics of germ tube formation in the null mutant were unaffected . Aberrant expression of HYR1 in yeast cells, when an ADH1-HYR1 fusion was used, did not stimulate hyphal formation in C . albicans or pseudohyphal growth in Saccharomyces cerevisiae . HYR1 appears to encode a nonessential component of the hyphal cell wall.

Scand J Immunol, 1996 Sep, 44(3), 204 - 14
Activity inhibition of cytolytic lymphocytes by omeprazole; Scaringi L et al.; This study examined the in vitro effect of omeprazole (OM) on various types of murine cytocidal lymphocytes . The results show that OM caused a strong inhibition of basal natural killer (NK) activity in spleen cells (SC) from untreated CD2F1 mice; in peritoneal exudate cells and SC activated in vivo by injection of maleic anhydride divinyl ether 1,2-copolymer (MVE-2) or inactivated Candida albicans (CA); in lymphokine-activated killer (LAK) activity generated in vitro from splenocytes cultured with rhIL-2 and in allo-specific cytotoxic lymphocyte-mediated lysis generated in vitro . A significant inhibition of cytotoxic activity of all types of effector cells after 30 min incubation was already induced by OM at 1 x 10(-3) M concentration, after 1 h incubation at 5 x 10(-4) M and after 4 h incubation at 1 x 10(-4) M OM . Complete inhibition of lytic activity was obtained after 4 h incubation of effector cells with 1 x 10(-3) M OM . No inhibitory effect was observed at 5 x 10(-5) M OM concentration . Indomethacin did not abrogate the OM inhibitory effect on NK/LAK activity, suggesting that prostaglandins are not involved in the process leading to suppression of cytocidal activity . When effector cells were incubated with OM in presence of rhIL-2 (500 U/ml), the cytokine failed to antagonize the inhibitory effect of the drug . On the contrary, if OM pretreated cells were incubated with rhIL-2 for a further 18 h after drug removal, this cytokine was able to restore NK activity, but only when NK inhibition was incomplete . These results demonstrate for the first time that in vitro OM causes a rapid, strong effect on various types of cytotoxic lymphocytes ranging from cytotoxicity inhibition to irreversible cell damage.

Curr Genet, 1996 Sep, 30(4), 325 - 31
Molecular cloning and expression of the nag1 gene (N-acetyl-beta-D-glucosaminidase-encoding gene) from Trichoderma harzianum P1; Peterbauer CK et al.; A 72-kDa N-acetyl-beta-D-glucosaminidase was purified from the mycoparasitic fungus Trichoderma harzianum P1; antibodies were raised against it, and aa-sequences were obtained . The antibody reacted with a single 72-kDa protein band in culture filtrates of T . harzianum grown on chitin, and was subsequently used to clone the corresponding nag1 gene from a lambdagt11 cDNA expression library . It was interrupted by two short introns and encoded a protein of 580 amino acids . The deduced protein sequence contained aa-sequence areas of high similarity to N-acetyl-glucosaminidases from other eukaryotes such as Candida albicans, and invertebrate and vertebrate animal tissues . The highest similarity was observed with the corresponding gene from the silkworm . The aa-sequence of a tryptic fragment of purified N-acetyl-beta-D-glucosaminidase from T . harzianum corresponded to a deduced aa sequence from a portion of the cloned gene, thus verifying that the protein is encoded by nag 1 . Southern analysis showed that nag 1 is present as a single-copy gene in T . harzianum . Expression of nag1-mRNA was strongly induced upon growth on chitin, N-acetyl-glucosamine and the cell walls of Botrytis cinerea used as a carbon source . The appearance of the corresponding N-acetyl-beta-D-glucosaminidase protein, as determined by Western analysis, paralleled the pattern of nag 1 expression, thereby suggesting that its formation is regulated at the level of transcription.

J Infect Dis, 1996 Sep, 174(3), 589 - 97
B cell knockout mice are resistant to mucosal and systemic candidiasis of endogenous origin but susceptible to experimental systemic candidiasis; Wagner RD et al.; Germfree J(H)D mice, which lack functional B cells and antibodies, were as resistant to orogastric and disseminated candidiasis of endogenous origin as were immunocompetent controls . Newborn J(H)D mice, in contrast to adult mice, were resistant to alimentary tract colonization by Candida albicans for 5-7 days after birth . C . albicans-colonized J(H)D mice were more resistant to intravenous challenge with C . albicans and had greater splenocyte proliferative responses to C . albicans antigens than did germfree mice or conventional controls . Thus, innate and acquired T cell-mediated immune responses induced after oral immunization are sufficient to protect J(H)D mice from mucosal and systemic candidiasis of endogenous origin; however, functional B cells may be required to protect mice from a primary intravenous challenge with C . albicans.

J Immunol, 1996 Sep 1, 157(5), 2155 - 9
TNF receptors in murine Candida albicans infection: evidence for an important role of TNF receptor p55 in antifungal defense; Steinshamn S et al.; TNF mediates multiple biologic activities through two distinct cell surface receptors, TNFR-p55 and TNFR-p75 . TNF plays an important role in nonspecific resistance against the fungus Candida albicans . We used transgenic mice deficient for TNFR-p55 or TNFR-p75 to investigate the role of the TNFR in antifungal defense . Mice deficient for TNFR-p55 have highly impaired ability to clear infection with C . albicans and readily succumb to the infection . Also mice deficient for TNFR-p75 had a significant reduction in their ability to clear the fungus although lethality was not increased . These data demonstrate that TNFR-p55 in particular, but also TNFR-p75, plays a definite role in defense against infection with C . albicans . In NMRI mice, infection with C . albicans resulted in a significant systemic release of soluble (s)TNFR-p75 . Cyclophosphamide-induced granulocytopenia led to a reduction of sTNFR-p75 release, whereas levels of bioactive TNF in response to fungal infection were increased . Release of sTNFR-p55 was not affected by induction of granulocytopenia . These observations suggest that granulocytes are a source of sTNFR-p75, possibly contributing to regulation of TNF activity during infection with C . albicans.

Infect Immun, 1996 Sep, 64(9), 3793 - 9
T lymphocytes in the murine vaginal mucosa are phenotypically distinct from those in the periphery; Fidel PL Jr et al.; The results from both clinical studies of women with recurrent vulvovaginal candidiasis and a murine model of experimental vaginitis indicate that systemic cell-mediated immunity may not represent a dominant host defense mechanism against vaginal infections by Candida albicans . Recent experimental evidence indicates the presence of local vaginal immune reactivity against C . albicans . The present study was designed to examine T-lymphocyte subpopulations in the vaginal mucosae of naive CBA/J mice . Vaginal lymphocytes (VL) were isolated by collagenase digestion of whole vaginal tissues . Cell populations were identified by flow cytometry, and the results were compared with those for both lymph node cells (LNC) and peripheral blood lymphocytes (PBL) . The results of flow cytometry showed that 45% +/- 10% of lymphocytes in the vaginal mucosa are CD3+ compared with 75% +/- 5% in LNC and 50% +/- 5% in PBL . The majority (85%) of CD3+ VL are CD4+ and express the alpha/beta T-cell receptor (TCR), similar to the results for LNC and PBL . In contrast to LNC and PBL, VL contain a significantly higher percentage (15 to 20%) of gamma/delta TCR+ cells, 80% or more of which appear to express CD4 . In addition, while CD4-CD8 cell ratios in LNC and PBL were 3:1 and 6:1, respectively, only 1% of VL expressed CD8, resulting in a CD4-CD8 cell ratio of > 100:1 . Finally, while LNC and PBL recognized two epitope-distinct (GK 1.5 and 2B6) anti-CD4 antibodies, VL recognized only 2B6 anti-CD4 antibodies . Further analysis of VL showed that Thy-1 cells, but not CD4 cells, were reduced after intravaginal injection of complement-fixing anti-Thy-1.2 and GK 1.5 anti-CD4 antibodies, respectively . Taken together, these data suggest that T lymphocytes in the vaginal mucosae of mice are phenotypically distinct from those in the periphery and that CD4+ VL have an uncharacteristic or atypical expression of the CD4 receptor.

Infect Immun, 1996 Sep, 64(9), 3752 - 7
Binding of resting platelets to Candida albicans germ tubes; Robert R et al.; The binding of resting platelets to Candida albicans germ tubes was studied by means of an affinity column in which germ tubes were physically immobilized . Adhesion of platelets to the column was dependent on both the germ tube concentration and the number of platelets applied . It was found that the interaction of C . albicans germ tubes with platelets is specific and should be mediated by a fungal protein receptor . The results obtained by scanning electron microscopy confirmed that resting platelets can fix directly onto germ tubes . In addition, this study showed that attachment of platelets onto C . albicans is associated with morphological changes . Platelets lost their discoid shape, became globular, generated spikes or pseudopods, and then flattened on the yeast cells.

Fertil Steril, 1996 Sep, 66(3), 481 - 3
Yeast infection of sperm, oocytes and embryos after intravaginal culture for embryo transfer; Mahadevan MM et al.; OBJECTIVE: To report a case of an embryo culture infected with Candida albicans after intravaginal culture . DESIGN: Case report . SETTING: Private infertility practice and university medical center . PATIENT: A couple with tubal and male factor infertility . INTERVENTIONS: Superovulation, oocyte recovery, Percoll sperm preparation, and intravaginal culture of sperm and oocytes in a tissue culture tube . MAIN OUTCOME MEASURES: Yeast infection of sperm, oocyte, and embryo culture . RESULTS: Candida albicans infection occurred in the sperm, oocyte, and embryo culture when cultured in a sealed tube in the vagina . Candida albicans also was found in the prepared sperm suspension culture in a separate tube kept in a 37 degrees C incubator . CONCLUSIONS: Infection of the embryo culture with C . albicans probably occurred when contaminated sperm was added at the time of insemination . Sperm preparation by the Percoll gradient centrifugation failed to eliminate C . albicans in the semen.

Eur J Biochem, 1996 Aug 15, 240(1), 37 - 44
Characterization of alpha-1,6-mannosyltransferase responsible for the synthesis of branched side chains in Candida albicans mannan; Suzuki A et al.; A particulate insoluble fraction from Candida albicans NIH B-792 (serotype B) strain cells was obtained as the residue after extracting a 105000 x g pellet of cell homogenate with 1% Triton X-100 . Incubation of this fraction with a mannopentaose, Man alpha 1-->3Man alpha 1-->2Man alpha 1-->Man alpha 1-->2Man, in the presence of GDP-mannose and Mn2+ at pH 6.0 gave a branched mannohexaose, {sequence: see text} 6 the structure of which was identified by means of sequential off assignment . However, the enzyme fraction obtained from Candida parapsilosis gave Man alpha 1-->2Man alpha 1-->3Man alpha 1-->2Man alpha 1-->2 Man alpha 1-->2Man under the same conditions . These results demonstrate the finding that the structural difference in the mannans of these two species is due to the presence of alpha-1.6-linked branching mannose units in the C . albicans mannan {Shibata, N., Ikuta, K., Imai, T., Satoh, Y., Satoh, R., Suzuki, A., Kojima, C., Kobayashi, H., Hisamichi, K . & Suzuki, S . (1995) J . Biol . Chem . 270, 1113-1122} . The substrate-specificity study of the enzyme indicated that the structural requirement of the alpha-1,6-mannosyltransferase is Man alpha 1-->3Man alpha 1--> . The alpha-1,6-mannosyltransferase also transferred the alpha-1,6-linked branching mannose unit to the mannan of Saccharomyces cerevisiae . The transformation of the mannan was detected by the appearance of antigenic factor 4 using an enzyme-linked immunosorbent assay and two-dimensional homonuclear Hartmann-Hahn spectroscopy.

FEMS Microbiol Lett, 1996 Aug 15, 142(1), 117 - 22
Expression of the fibrinogen binding mannoprotein and the laminin receptor of Candida albicans in vitro and in infected tissues; Lopez-Ribot JL et al.; We have previously reported a 37 kDa laminin-binding protein (p37) and a 58 kDa fibrinogen-binding mannoprotein (mp58) on the surface of Candida albicans . A few yeast cells expressed both functional receptors at the surface while germ tubes expressed a functional mp58 fibrinogen but not a functional p37 laminin receptor . These receptors were heterogeneously dispersed at the surface as shown by binding of rabbit antiserum to mp58 (PAb anti-mp58) and antiserum to the human high affinity laminin receptor . In this report we have used a dual fluorescence technique to determine if the two receptors colocalize, perhaps as part of a receptor complex . Fibrinogen was used as a probe for mp58 and polyclonal antiserum generated to the p37 (PAb anti-p37) was used as a probe for the 37 kDa laminin-binding protein . Both receptors were heterogeneously distributed, but the receptors were not colocalized as the areas of concentration of each receptor were different . Immunohistochemical analysis of tissue sections from patients with disseminated and superficial candidiasis with PAb anti-p37 and PAb anti-mp58 revealed that both receptors were also expressed in infected tissues . The patterns of morphological expression were similar to the in vitro patterns detected by immunofluorescence.

J Acquir Immune Defic Syndr Hum Retrovirol, 1996 Aug 15, 12(5), 470 - 6
Prophylaxis with nystatin pastilles for HIV-associated oral candidiasis; MacPhail LA et al.; To determine whether daily use of nystatin pastilles can prevent initial outbreak or recurrence of oral candidiasis in HIV-infected patients and to identify factors associated with outbreaks during 20-week follow-up, a randomized, double-blind, placebo-controlled clinical trial was conducted . Subjects were 128 HIV-infected men (aged 27-60 years) who either had had no documented episode of oral candidiasis in the previous year or had been clinically clear of oral candidiasis for at least 72 h before randomization . Study arms were two placebo pastilles, one nystatin (200,000 U) and one placebo pastille, or two nystatin pastilles daily for 20 weeks . The main outcome measure was time to oral candidiasis, as determined by potassium hydroxide (KOH) smear and fungal culture . A multivariate proportional hazards model showed that four factors were significant (p < 0.001) in predicting time to oral candidiasis: nystatin treatment (hazard ratio 0.59), history of oral candidiasis (3.58), Candida albicans carriage (2.79), and CD4 count at randomization (0.65) . In this small group of subjects, nystatin appeared to be effective in delaying onset of oral candidiasis . Patients with CD4 counts < 200 who are carriers of C . albicans and have a history of oral candidiasis may be most likely to benefit from antifungal prophylaxis.

Biochem Biophys Res Commun, 1996 Aug 5, 225(1), 47 - 53
Stabilization of helix by side-chain interactions in histatin-derived peptides: role in candidacidal activity; Ramalingam K et al.; Candida albicans is an opportunistic pathogen prevalent in AIDS patients and oral candidiasis . Azolebased drugs are currently used in the treatment of candidiasis . Histidine-rich peptides (histatins), are the natural inhibitors of candida species present in human salivary secretions . Sequence comparison of histatins revealed the common motif--KRKFHE--in active peptide fragments . Molecular modeling analysis showed structural similarity between this segment of histatins and azole-based drugs . The helical conformation adopted by histatin-5 may be stabilized by two side chain-side chain interactions (Phe.. . His and Arg .. . Glu) . Based on sequence comparison of histatin peptides and molecular modeling, a synthetic 10-residue peptide derived from histatin-5 was helical and possessed significant anti candida activity . This peptide may be used as a template to develop histatin-based drugs for treating oral candidiasis.

Genes Cells, 1996 Aug, 1(8), 727 - 40
Analysis of the cell cycle in the budding yeast Candida albicans by positioning of chromosomes by fluorescence in situ hybridization (FISH) with repetitive sequences; Chibana H et al.; BACKGROUND: In the budding yeasts, including Saccharomyces cerevisiae, in which individual chromosomes cannot be visualized by microscopy, the mitotic phases in the cell cycle have not been correlated with the chromosome behaviour . We used various repetitive sequences, namely, rDNA, telomeric sequences and RPSs, which are localized in limited regions in almost all chromosomes, as probes for fluorescence in situ hybridization (FISH) to analyse the cell cycle phases in a pathogenic yeast Candida albicans . The positioning of the FISH signals was analysed quantitatively in relation to the length of spindle microtubules in the nuclear domain . RESULTS: RPSs were randomly distributed in the interphase nucleus, and they formed aggregates with the development of the spindle . DNA synthesis was complete before RPSs came closest to the spindle . As the spindle elongated, they were scattered along the spindle and then separated into two clusters at the spindle poles at the end of anaphase . rDNA was localized in the nucleolar domain, and telomere signals were randomly distributed throughout mitosis . CONCLUSION: By estimating quantitatively the proportions of mitotic cells with particular configurations of both microtubules and chromosomes in a population of rapidly proliferating cells, we were able to define various stages in the progression of mitosis . The S phase and pro-to-prometaphase were overlapping and the G2 phase was lacking . Unexpectedly, the pole-to-pole elongation of the spindle (anaphase B) was predominating and was followed by movement of chromosomes to the poles (anaphase A).

Jpn J Antibiot, 1996 Aug, 49(8), 818 - 23
{In vitro antifungal activity of omoconazole nitrate, a novel imidazone antimycotic drug, against clinical isolates from patients with cutaneous mycosis}; Uchida K et al.; In vitro antifungal activities of omoconazole nitrate (OMZ), a novel antifungal imidazole antimycotic drug, are examined against clinical isolates obtained from patients with cutaneous mycosis and its activity was compared with that of bifonazole (BFZ) . The clinical isolates tested were 70 of dermatophytes including Trichophyton rubrum (47 isolates), T . mentagrophytes (22 isolates), Microsporum gypseum (1 isolate), and 27 isolates of Candida albicans . MIC values of OMZ to dermatophytes distributed in a range of < or = 0.04 to 0.63 microgram/ml were similar to those of BFZ (< or = 0.04 to 1.25 micrograms/ml) . MIC values of OMZ to C . albicans were in a range of 0.16 to 2.5 micrograms/ml indicating that OMZ had more potent activities than BFZ (1.25 to 5 micrograms/ml) . These results showed that in vitro antifungal activities of OMZ against clinical isolates of dermatophytes and C . albicans were greater than or similar to those of BFZ.

Arch Dermatol Res, 1996 Aug, 288(9), 495 - 9
Reduced proliferative responses of peripheral blood mononuclear cells specifically to Candida albicans antigen in patients with atopic dermatitis--comparison with their normal reactivity to bacterial superantigens; Tanaka M et al.; Although reduced cutaneous reactivities to Candida albicans have been reported in patients with atopic dermatitis (AD), there is still controversy as to whether the in vivo lymphocyte proliferation response is normal or reduced . We have also reported that patients with AD manifest a decreased cutaneous response only to C . albicans antigen in scarification patch tests . The purpose of this study was to examine whether patients with AD show normal lymphocyte transformation responses to C . albicans antigen . Peripheral blood leucocytes (PBL) from 21 patients with AD and 14 healthy control (HC) subjects were cocultured with optimal concentrations of C . albicans antigen and of superantigens (staphylococcal enterotoxin A and B) . PBL from the patients with AD showed a significantly lower response to C . albicans antigen, but there was no statistically significant difference in PBL responses to superantigens between the patients with AD and the HC subjects . This decreased proliferative response of PBL was particularly noticeable in those whose RAST scores for C . albicans antigen were high . These results suggest the development of a specific anergy to C . albicans antigen in patients with AD.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1996 Aug, 82(2), 161 - 5
Oral pathoses caused by Candida albicans during chemotherapy: update on development mechanisms; Bunetel L et al.; Oral candidiasis occurs at a high frequency among immunocompromised hosts . The development mechanisms of oral pathoses associated with Candida are complex and certainly multifactorial . In immunocompromised patients, they include the evolution of the buccal flora associated with the influence of antineoplastic treatments and immunosuppression . They also include adherence of Candida to epithelial cells of the oral cavity as a function of host cell-related and yeast-related factors . Interaction and cooperation between Candida and bacteria could be a third influence in the development of oral candidiasis . It seems important to determine these mechanisms more precisely so as to improve preventive and therapeutic measures.

J Dermatol, 1996 Aug, 23(8), 572 - 6
Pruritic papular eruptions and candidiasis due to HIV infection; Uchigasaki S et al.; We present two patients with refractory papular eruptions and severe candidiasis . Both of them are positive for treponema pallidum and have suffered from pruritic papular eruptions (PPE) that had resisted therapy for years . Also, candidiasis appeared in the mouth, at intertriginous sites, and on the feet . The clinical features suggested immunodeficiency, and HIV tests were positive . Histologically, the specimen from the PPE lesion showed perivascular and perifollicular mixed cell infiltration . The fungus was identified by both Parker-KOH-mount examination and mycologic culture as Candida albicans . The pruritic papules were healed almost completely with oral antihistamine and topical corticosteroid treatment, and the candidiasis mostly disappeared after treatment with topical antifungal agents alone . We learned from these two cases that refractory PPE and severe candidiasis indicate a need for HIV testing.

AIDS, 1996 Aug, 10(9), 983 - 7
Dysregulation of the polymorphonuclear leukocyte--Candida spp . interaction in HIV-positive patients; Wenisch C et al.; OBJECTIVE: In HIV-infected patients there is an increased frequency of fungal infections . Dysregulation of the response of phagocytic cells to fungal pathogens may be involved . DESIGN: Phagocytosis of Candida spp., consecutive intracellular production of reactive oxygen species, and candicidal activity were analysed in polymorphonuclear leukocytes (PML) from HIV-1-infected patients, who were at stage C3 of the 1993 revised Centers for Disease Control and Prevention classification system, by means of flow cytometry . METHODS: Phagocytic ability was assessed by measuring uptake of fluorescein isothiocyanate-labelled Candida albicans, C . krusei and C . glabrata . Reactive oxygen intermediate production was estimated by the quantity of dihydrorhodamine-123 converted to rhodamine-123 intracellularly . The candicidal effect was assessed by the propidium iodide uptake of killed yeast cells . RESULTS: As compared to PML of healthy, HIV-negative controls, PML of AIDS patients exhibited an increased phagocytic activity and a similar ability to generate reactive oxygen products . In contrast, PML of AIDS patients displayed a decreased candicidal activity (P < 0.05 compared to controls) . CONCLUSION: These results suggest that in patients with advanced HIV-1 infection the impairment of non-oxidative killing mechanisms of phagocytic cells may contribute to the high incidence of fungal infections.

Antimicrob Agents Chemother, 1996 Aug, 40(8), 1806 - 10
Evaluation of renal toxicity and antifungal activity of free and liposomal amphotericin B following a single intravenous dose to diabetic rats with systemic candidiasis; Wasan KM et al.; Since fungal infections are prevalent in diabetic patients, in whom treatment is often complicated by underlying renal disease and dyslipidemias, the purpose of the present study was to determine if the antifungal activity and nephrotoxic effects of amphotericin B (AmB) and liposomal AmB (L-AmB) are different in nondiabetic (normolipidemic) rats compared with those in diabetic (dyslipidemic) rats with systemic candidiasis . Non diabetic and diabetic rats infected with Candida albicans received a single intravenous dose of either AmB (0.8 mg of AmB per kg of body weight), L-AmB (0.8, 2, or 4 mg of AmB per kg), or an equivalent volume of normal saline (1 ml) . Renal function was assessed by insulin clearance, and antifungal activity was determined by measuring the numbers of CFU of C . albicans that were present in the right kidney following drug treatment . AmB at 0.8 mg/kg and L-AmB at 0.8, 2, and 4 mg/kg are effective antifungal agents in both diabetic and nondiabetic rats . However, while there was approximately a 4-fold decline in the mean number of CFU per gram of kidney in nondiabetic rats, there was only approximately a 2.5-fold decline for the comparable dose (AmB, 0.8 mg/kg) in diabetic rats . There also appeared to be a similar fold reduction of L-AmB at all of the dosages tested . AmB treatment significantly improved renal function in diabetic and nondiabetic rats with systemic candidiasis . Although L-AmB at all doses tested significantly improved renal function in diabetic rats with systemic candidiasis, only L-AmB at doses of 2 and 4 mg/kg significantly improved renal function in nondiabetic rats with systemic candidiasis . These findings suggest that following administration of a single intravenous dose, AmB and L-AmB appear to be less effective in killing C . albicans isolates in diabetic than in nondiabetic rats, while they were found to improve the renal functions of rats in both treatment groups.

J Clin Microbiol, 1996 Aug, 34(8), 2039 - 41
Colorimetric susceptibility testing for Aspergillus fumigatus: comparison of menadione-augmented 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide and Alamar blue tests; Jahn B et al.; Two colorimetric methods that use Alamar Blue or 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) for assaying the in vitro activities of antifungal agents have been described . We report that both tests performed similarly when the antifungal activity of amphotericin B against Candida albicans was determined . However, only the MTT test generated interpretable data when Aspergillus fumigatus was used.

Planta Med, 1996 Aug, 62(4), 308 - 13
Antifungal activity from Alpinia galanga and the competition for incorporation of unsaturated fatty acids in cell growth; Haraguchi H et al.; An antimicrobial diterpene was isolated from Alpinia galanga in the screening for potentiators of phytochemical antibiotic action . The structure was elucidated by spectral data and identified as (E)-8 beta, 17-epoxylabd-12-ene-15, 16-dial (1) . Diterpene 1 synergistically enhanced the antifungal activity of quercetin and chalcone against Candida albicans . Antifungal activity of 1 was reversed by unsaturated fatty acids . Protoplasts of C . albicans were lysed by 1 . These results suggest that antifungal activity of 1 is due to a change of membrane permeability arising from membrane lipid alternation.

DNA Cell Biol, 1996 Aug, 15(8), 617 - 24
Uptake of recombinant myeloperoxidase, free or fused to Fc gamma, by macrophages enhances killing activity toward micro-organisms; Tournay C et al.; A chimeric antibody-like molecule consisting of the human myeloperoxidase (rMPO) fused to the second and third constant-sequence (CH2 and CH3) Fc domains of human immunoglobulin G-1 has been constructed and expressed in Chinese hamster ovary (CHO) cells . This fusion molecule was designed to combine the binding specificity of Fc with the antimicrobial properties of rMPO . The rMPO-Fc fusion dimerized through the Fc fragment, while retaining the enzymatic activity of rMPO . The chimeric molecule was glycosylated and most of the propeptide was eliminated, indicating a better processing of the polypeptide than for rMPO alone . Both rMPO and rMPO-Fc bound to and were internalized by macrophage-like U937 promonocytic cells . Unexpectedly, the chimera failed to bind to the Fc receptor but interacted with a higher affinity than rMPO with the same binding sites . The presence of the Fc fragment in the chimera, in addition, did not extend the plasma half-life of the fusion protein . In vitro, rMPO-Fc exhibited a stronger killing effect than rMPO toward Candida albicans in the presence of either H202 alone or human macrophages . In vivo, rMPO-Fc similarly conferred a better protection than rMPO in a lethal model of murine cowdriosis . These properties could be related to the Fc-induced dimerization of the fusion protein in CHO cells.

J Pediatr, 1996 Aug, 129(2), 245 - 50
Delayed-type hypersensitivity skin testing in human immunodeficiency virus-infected pediatric patients; Raszka WV et al.; OBJECTIVE: To evaluate whether pediatric patients infected with human immunodeficiency virus (HIV) can mount appropriate delayed-type hypersensitivity (DTH) skin responses to recall antigens and whether these responses can be correlated with clinical or immunologic parameters . DESIGN: Prospective evaluation of DTH responses in HIV-infected children . Uninfected children born to HIV-infected mothers served as control subjects . Antigens used for yearly DTH testing included Candida albicans (1:100, 1:10); mumps virus; Trichophyton; purified protein derivative of tuberculin; and tetanus toxoid (1:100, 1:10) . At the time of each DTH test, patients were staged according to two Centers for Disease Control and Prevention pediatric HIV classification systems, and T-cell subsets were obtained . RESULTS: Twenty-seven HIV-infected patients with a median age at entry of 74.1 (range, 12 to 156) months were followed . Forty-four DTH skin tests in 21 symptom-free HIV-infected patients (PI) and 18 tests in 10 HIV-infected patients with symptoms (P2), as well as 43 DTH skin tests in 18 patients who had either mild or moderate clinical symptoms or immunosuppression and 19 tests in 13 patients with severe symptoms or immunosuppression, were evaluated . Sixteen DTH skin tests were performed in 14 uninfected patients . HIV-infected patients tended to have fewer DTH responses to antigens and of smaller size than did uninfected patients . When controlled for age, few differences in DTH responsiveness were seen between HIV-infected and uninfected patients . Anergy was associated with symptomatic disease, evidence of advanced clinical or immunologic disease, and low CD4+ percentages (p <0.05) . CONCLUSIONS: HIV-infected children are able to mount antigen-specific cell-mediated immune responses that are qualitatively similar to those of age-matched control subjects . Loss of DTH responsiveness correlates with both clinical and immunologic evidence of HIV disease progression.

Microbiology, 1996 Aug, 142 ( Pt 8), 2271 - 7
Effect of monoclonal antibodies directed against Candida albicans cell wall antigens on the adhesion of the fungus to polystyrene; San Millan R et al.; The adhesion of Candida albicans to polystyrene and the effect of three monoclonal antibodies (mAbs) reactive with C . albicans cell wall surface antigens on this process was assessed in vitro with several C . albicans strains . In the absence of mAbs, adhesion of C . albicans to polystyrene increased in parallel with germ-tube formation . However, the growth of the strains in the yeast phase at 25 degrees C or the use of an agerminative mutant inhibited adhesion to polystyrene . Serotype A and B strains showed similar kinetics of adhesion to polystyrene and no statistically significant differences in germination or adhesion were observed when strains from the two serotypes were compared . The three mAbs had different effects on both germination and adhesion of C . albicans . mAbs 3D9 showed no influence on either germination or adhesion to polystyrene in two C . albicans strains . mAb B9E decreased both adhesion (45.6%) and filamentation (52.6%), and mAb 21E6 decreased filamentation (34.0%) but enhanced adhesion by 23.3% . This enhancement was also observed with the agerminative mutant and it was dose-dependent . It was not related to the binding capacity of the MAb to polystyrene nor to an increase in cell surface hydrophobicity of the antibody-treated cells . In conclusion, both growth phases of C . albicans can adhere to polystyrene, although the conditions for this process seem to be different in each phase . The two types of adhesion of C . albicans to polystyrene might have a role in the colonization of medical implants . The disparate effects shown by mAbs directed against cell wall mannoproteins of C . albicans on the adhesion of the fungus to polystyrene should be taken into consideration when designing strategies to block the adhesion of C . albicans to plastic materials with mAbs.

Microbiology, 1996 Aug, 142 ( Pt 8), 2263 - 70
Evidence for different mannosylation processes involved in the association of beta-1,2-linked oligomannosidic epitopes in Candida albicans mannan and phospholipomannan; Trinel PA et al.; A monoclonal antibody specific for beta-1,2-linked oligomannosides was used to study the association of these residues with Candida albicans mannan and phospholipomannan (PLM) in relation to growth conditions and in mannan mutant strains . Double immunofluorescence assays performed on cells grown under standard conditions indicated a highly heterogeneous cell surface expression of these epitopes in comparison with the homogeneous expression of alpha-linked oligomannosidic epitopes . Growth in the presence of tunicamycin, which inhibits mannan N-glycosylation, resulted in an absence of beta-1,2-oligomannosidic epitopes on the cell surface, although PLM synthesis still occurred as shown by autoradiography . Similarly, growth in acidic conditions, which inhibits the incorporation of beta-1,2-oligomannosides in mannan, resulted in an absence of beta-1,2-oligomannosidic epitopes at the cell surface, although they still associated with PLM as shown by Western blotting . Western blots of C . albicans mutant strains with reduced amounts or an absence of phosphorus and acid-labile beta-1,2-oligomannosides in their mannan confirmed that the association of beta-1,2-linked oligomannosides with mannan and with PLM involves different mannosylation processes.

Microbiology, 1996 Aug, 142 ( Pt 8), 2255 - 62
A comparative study of the incorporation of a 1,6-beta-glucan and an O-glycosylated protein epitope into the cell wall of Candida albicans; Sanjuan R et al.; The topological distribution of two epitopes in the cell wall of Candida albicans, the kinetics of their incorporation into the regenerating protoplast wall, and the effect of different antibiotics upon their incorporation and localization have been studied . To do so, two monoclonal antibodies that react against an O-glycosylated mannoprotein (1B12) and against a 1,6-beta-glucan epitope (JRR1) were used . The results show that the JRR1 epitope is localized in an internal layer of the cell wall, in contrast to the 1B12 epitope, which is superficial, and that the incorporation of the JRR1 epitope into walls of regenerating protoplasts precedes that of the 1B12 epitope . The JRR1 epitope is normally found in the culture medium of control cells, but not in that of papulacandin-B-treated cells, and tunicamycin interferes with the incorporation of the 1B12 epitope into the cell walls . Finally, the results support the hypothesis that mannoproteins are not 1,6-beta-glycosylated before their secretion.

Microbiology, 1996 Aug, 142 ( Pt 8), 2245 - 54
Cytoplasmic localization of the white phase-specific WH11 gene product of Candida albicans; Schroppel K et al.; Cells of Candida albicans WO-1 switch frequently, spontaneously and reversibly between a white and opaque phase . The white-opaque transition involves the regulation of phase-specific genes . In the white budding phase, cells express the white phase-specific gene WH11, which encodes a protein with homology to the heat shock protein Hsp12 of Saccharomyces cerevisiae . A recombinant Wh11 protein has been synthesized, purified to apparent homogeneity and used to generate a rabbit polyclonal antiserum . The antiserum was used to localize the Wh11 protein in white phase cells . Wh11 is distributed throughout the cytoplasm but appears to be excluded from vesicles, plasma membrane and nucleus . An analysis by Western blotting of Wh11 expression in a number of C . albicans strains and related species suggests a correlation between round budding cell shape and expression.

Infect Immun, 1996 Aug, 64(8), 3333 - 40
Evidence for presence in the cell wall of Candida albicans of a protein related to the hsp70 family; Lopez-Ribot JL et al.; We have previously reported the isolation of several clones from a cDNA expression library from Candida albicans, one of which was associated with a constitutively expressed 70-kDa protein . The moiety was present in the beta-mercaptoethanol extracts of cell walls from both blastoconidia and germ tubes . The surface expression of this moiety was revealed by an indirect immunofluorescence assay using affinity-purified antibody to the fusion protein produced by the clone . The 0.68-kb cDNA insert was sequenced . A database search revealed extensive homology with the 70-kDa family of stress or heat shock proteins (hsps) . The 77% homology with another C . albicans HSP70 sequence suggested that this fragment represented a second member of the HSP70 family in this organism . Homology ranging from 65 to 76% was observed with members of four subfamilies (SSA, SSB, SSC, and SSD) of the Saccharomyces cerevisiae HSP70 gene family . The nucleic acid sequence and the deduced amino acid sequence of the open reading frame showed greatest homology with SSA1 and SSA2 sequences, and the gene corresponding to the cDNA clone was designated C . albicans SSA2 . The relationship with the SSA family was supported by reactivity of the 70-kDa component with antibody recognizing the Ssa proteins of S . cerevisiae . The presence of an hsp70 in the cell wall was confirmed by two additional methods . Cell wall proteins were biotinylated with a non-membrane-permeable derivative to distinguish extracellular from cytosolic proteins . Biotinylated hsp70 was detected by Western blotting (immunoblotting) among the biotinylated components affinity purified by chromatography on streptavidin, thereby establishing its presence in the cell wall . Immunoelectron microscopy showed that the 70-kDa component was present at the cell surface as well as the outer surface of the plasma membrane and extended through the cell wall, occasionally appearing to reach the cell surface through channels . Northern (RNA) blot analysis showed that the gene was expressed in yeast cells growing in yeast extract-peptone medium at both 25 and 37 degrees C and in Lee medium at 25 degrees C and during formation of germ tubes in Lee medium 37 degrees C . No obvious increase in the expression level was detected after the temperature shift . Members of the hsp70 family have been reported to be immunoreactive . The fusion protein produced by the cDNA clone was recognized by serum from healthy individuals and patients with candidiasis . Since members of the hsp70 family of eucaryotic proteins are associated with chaperone and translocation functions, in addition to being immunogenic, this protein may play a role in the assembly and function of other cell wall proteins.

Infect Immun, 1996 Aug, 64(8), 3244 - 51
Characterization of echinocandin-resistant mutants of Candida albicans: genetic, biochemical, and virulence studies; Kurtz MB et al.; The pneumocandins are potent antifungal agents of the echinocandin class which are under development for use as broad-spectrum antimycotic therapy . One important consideration for any new therapeutic class for treating serious fungal infections is the potential for drug resistance development . In this study we have isolated and characterized four independent spontaneous Candida albicans mutants resistant to the potent semisynthetic pneumocandin L-733,560 . These mutants have many of the properties of FKS1/ETG1 echinocandin-resistant mutants of Saccharomyces cerevisiae, including (i) cross-resistance to other 1,3-beta-D-glucan synthase inhibitors, such as papulacandin and echinocandins, but no change in sensitivity to other antifungal agents; (ii) in vitro glucan synthase activity that is more resistant to pneumocandins than the wild-type parent enzyme; and (iii) semidominant drug resistance in spheroplast fusion strains . The mutants were compared with C . albicans echinocandin-resistant mutants isolated by mutagenesis by L . Beckford and D . Kerridge (mutant M-2) (abstr . PS3.11, in Proceedings of the XI Congress of the International Society for Human and Animal Mycology, Montreal, Canada, 1992) and by A . Cassone, R . E . Mason, and D . Kerridge (mutant CA-2) (Sabouraudia 19:97-110, 1981) . All of the strains had resistant enzyme activity in vitro . M-2 grew poorly and had low levels of enzyme activity . In contrast, CA-2 and the spontaneous mutants grew as well as the parents and had normal levels of glucan synthase activity . These results suggest that these resistant mutants may have alterations in glucan synthase . CA-2 was unable to form germ tubes, an ability retained by the spontaneous mutants . The virulence of the spontaneous mutants was unimpaired in a mouse model of disseminated candidiasis, while M-2 and CA-2 were 2 orders of magnitude less virulent than their parent strains . Significantly, mice challenged with the spontaneous mutant CAI4R1 responded therapeutically to lower levels of L-733,560 than would he predicted by the increase in in vitro susceptibility.

Infect Immun, 1996 Aug, 64(8), 3127 - 33
Peroxynitrite contributes to the candidacidal activity of nitric oxide-producing macrophages; Vazquez-Torres A et al.; Nitric oxide (NO) is associated with functions as diverse as peristalsis, blood flow, neuroendosecretion, visual transduction, smooth muscle relaxation, and microbial killing (H . H . W . H . Schmidt and V . Walter, Cell 78:919-925, 1994) . Despite the well-established role of NO in macrophage candidacidal activity (E . Cenci, L . Romani, A . Mancacci, R . Spaccapelo, E . Schiaffella, P . Puccetti, and F . Bistoni, Eur . J . Immunol . 23:1034-1038, 1993; J . Jones-Carson, A . Vazquez-Torres, H . Van der Heide, R . D . Wagner, T . Warner, and E . Balish, Nature Med . 1:552-557, 1995; and A . Vazquez-Torres, J . Jones-Carson, T . Warner, and E . Balish, J . Infect . Dis . 172:192-198, 1995), NO is not directly candidacidal for Candida albicans (A . Vazquez-Torres, J . Jones-Carson, and E . Balish, Infect . Immun . 63:1142-1144, 1995) . Because macrophages can produce both NO and superoxide anion (02-), we postulated that peroxynitrite (ONOO-), a product of the dilution-limited reaction of NO and O2-, is the candidacidal molecule of activated macrophages . We now report that ONOO-, in addition to being candidacidal in vitro, is responsible for the candidacidal activity of NO-producing macrophages . ONOO- synthesis by NO-producing macrophages was triggered by two independent mechanisms: one was nonopsonic and dependent on fungal cell wall glucan moieties, and the other was dependent on opsonic antibodies . As we have demonstrated for the pathogenic fungus C . albicans, ONOO- may also be the molecule that enables macrophages to kill other microbes that are resistant to both O2- and NO.

Infect Immun, 1996 Aug, 64(8), 2936 - 40
Induction of an extracellular esterase from Candida albicans and some of its properties; Tsuboi R et al.; An extracellular esterase from Candida albicans A-714 was found to be induced in a medium containing 0.7% yeast nitrogen base and 2.5% Tween 80 (polyoxyethylenesorbitan compounds) . Enzyme activity, which exists predominantly in the extracellular space, was measured by a colorimetric method using alpha-naphthyl palmitate as a substrate . The induction level of the esterase activity was found to be well correlated with fungal growth and was dependent on the Tween 80 concentration . Such esterase activity was observed only in medium containing Tween 80 or other Tweens as the sole carbon source and therefore was not observed in either peptone-glucose medium or peptone-glucose medium supplemented with Tween 80 . The induced esterase was heat labile and had maximum activity at pH 5.5 . Enzyme activity was stimulated by the addition of sodium taurocholate, an activator of lipase . Thin-layer chromatography revealed that this enzyme does not hydrolyze triolein and L-alpha-lecithin, suggesting that it is a monoester hydrolase (not a lipase in the strict sense of the word) . Esterase activity was examined in 85 clinical isolates of Candida species; C . albicans, C . tropicalis, and C . parapsilosis tended to have higher enzyme activities than C . kefyr, C . krusei, C . glabrata, and C . guilliermondii . Although the physiological properties of this esterase are not clear at present, it was found to be crucial for fungal growth under specific conditions.

Infect Immun, 1996 Aug, 64(8), 2930 - 5
Specific induction of fibronectin binding activity by hemoglobin in Candida albicans grown in defined media; Yan S et al.; Fibronectin (FN) is a major component of host extracellular matrix that may play an important role in the initiation and dissemination of Candida albicans infections . Expression of FN binding requires growth of C albicans blastoconidia in complex medium, and the regulation of FN receptor expression is poorly understood . We now demonstrate that hemoglobin is a potent and specific inducer of FN receptor expression and describe a defined medium supplemented with hemoglobin that greatly and stably enhances the binding activity of C . albicans for soluble FN . Enhancement of FN binding by hemoglobin in strain 44807 was concentration dependent and was maximal at 0.1% hemoglobin with 20- to 80-fold enhancement . The hemoglobin-induced FN binding to C . albicans was saturable, with a Kd of 2.7 X 10(-8) M . Enhancement required growth of C . albicans in hemoglobin-containing medium, since simply exposing blastoconidia to hemoglobin in a nongrowing status did not enhance binding . Induction was reversible following removal of hemoglobin from the growth medium and not associated with germination . Inorganic or protein-bound iron was not sufficient for the induction, since other iron-containing proteins or inorganic iron salts were inactive . Growth in the simple medium yeast nitrogen base supplemented with hemoglobin increased cell adhesion to immobilized FN and to cultured monolayers of bovine corneal endothelial cells . These data suggest that hemoglobin may be an important regulator of FN binding activity in C . albicans and thus may play a role in its pathogenesis.

J Bacteriol, 1996 Aug, 178(15), 4635 - 42
Beta-glucan synthesis in Bradyrhizobium japonicum: characterization of a new locus (ndvC) influencing beta-(1-->6) linkages; Bhagwat AA et al.; Bradyrhizobium japonicum synthesizes periplasmic cyclic beta-(1-->3),beta-(1-->6)-D-glucans during growth in hypoosmotic environments, and evidence is growing that these molecules may have a specific function during plant-microbe interactions in addition to osmoregulation . Site-directed Tn5 mutagenesis of the DNA region upstream of ndvB resulted in identification of a new gene (ndvC) involved in beta-(1--> 3), beta-(1-->6)-glucan synthesis and in nodule development . The predicted translation product was a polypeptide (ca . 62 kDa) with several transmembrane domains . It contained a sequence characteristic of a conserved nucleoside-sugar-binding motif found in many bacterial enzymes and had 51% similarity with a beta-glucanosyltransferase from Candida albicans . B . japonicum carrying a Tn5 insertion in ndvC resulted in synthesis of altered cyclic beta-glucans composed almost entirely of beta-(1--> 3)-glycosyl linkages . The mutant strain was only slightly sensitive to hypoosmotic growth conditions compared with the ndvB mutant, but it was severely impaired in symbiotic interactions with soybean (Glycine max) . Nodulation was delayed by 8 to 10 days, and many small nodule-like structures apparently devoid of viable bacteria were formed . This finding suggests that the structure of the beta-glucan molecule is important for a successful symbiotic interaction, and beta-glucans may have a specific function in addition to their role in hypoosmotic adaptation.

Crit Care Med, 1996 Aug, 24(8), 1311 - 5
Administration of amphotericin B in lipid emulsion decreases nephrotoxicity: results of a prospective, randomized, controlled study in critically ill patients; Sorkine P et al.; OBJECTIVES: To evaluate the differences in efficacy and in clinical and biochemical tolerance to amphotericin B administered in a lipid emulsion compared with amphotericin B administered in 5% dextrose in water in the treatment of Candida albicans infection in intensive care unit (ICU) patients . DESIGN: Prospective, controlled, randomized study, conducted during a 2.5-yr period, comparing the two treatment protocols . SETTING: General ICU of a university-affiliated municipal hospital . PATIENTS: Sixty consecutive critically ill patients with confirmed or suspected Candida infection . INTERVENTIONS: Patients received amphotericin B (1 mg/kg/24 hrs), administered randomly in 5% dextrose in water (group A), or in lipid emulsion (20% intralipid) (group B) . MEASUREMENTS AND MAIN RESULTS: Clinical tolerance (fever, chills, hemodynamics), hepatorenal tolerance, and biological tolerance (serum electrolytes and coagulation profile) were evaluated . Patients receiving amphotericin B in lipid emulsion experienced a lower frequency rate of drug-associated fever (61.4% vs . 5.8%, p < .003) rigors (54% vs . 8.5%, p < .004), hypotension (17% vs . 0%), and nephrotoxicity (increase of serum creatinine concentration 66.7% vs . 20%, p < .0002) . Significant (264,500 +/- 71,460 to 163,570 +/- 34,450 mm3, p < .01) thrombocytopenia, not associated with active bleeding, occurred in patients receiving amphotericin B lipid in emulsion but not in patients receiving the drug in dextrose . CONCLUSIONS: Treatment with amphotericin B in a lipid emulsion when given to critically ill patients with Candida sepsis seems to be safer and as effective as the conventional mode of administration.

J Infect Dis, 1996 Aug, 174(2), 435 - 9
Host defenses against disseminated candidiasis are impaired in intercellular adhesion molecule 1-deficient mice; Davis SL et al.; Genetically engineered mice, which lack normal expression of intercellular adhesion molecule 1 (ICAM-1), were used to study the role of ICAM-1 in the host defense against disseminated candidiasis . The responses of ICAM-1-deficient mice and normal wild type mice were compared following an intravenous challenge with Candida albicans . ICAM-1-deficient mice lost more weight (P < .001) and had a significantly higher mortality (P < .001) . Quantitative cultures revealed a greater tissue fungal burden in ICAM-1-deficient mice compared with normal mice, in both the kidney (P < .001) and the brain (P = .007) . Extensive inflammation, composed primarily of histiocytes admixed with lymphocytes and occasional neutrophils, was present in the renal tissue of ICAM-1-deficient mice; this contrasted with a more localized and predominantly neutrophilic infiltrate in normal mice . This work suggests that the loss of ICAM-1 significantly impairs host defense against C . albicans, by impairing either neutrophil migration or phagocyte activation or both.

Curr Genet, 1996 Jul 31, 30(2), 115 - 20
Characterization of the Saccharomyces cerevisiae FMS1 gene related to Candida albicans corticosteroid-binding protein 1; Joets J et al.; In order to investigate ergosterol metabolism in S . cerevisiae we studied the CM8 mutant strain defective in the regulation of this pathway . A genomic multicopy library was screened to reverse the CM8 phenotype . This allowed us to characterize a new gene, FMS1, which relieves mutant phenotype by extragenic functional complementation . FMS1 may encode a 508 amino-acid protein . The predicted protein shares 35% identity with Cbp1p, a Candida albicans corticosteroid binding-protein . Fms1p also shows a weaker homology with monoamine oxidases . The construction of a FMS1 null-allele yeast strain demonstrated that this gene is not essential for yeast in normal usual laboratory culture conditions . The existence of a gene related to CBP1 of C . albicans in S . cerevisiae strongly suggests a possible function of steroid-binding proteins in yeast general physiology rather than in a process related to pathogenicity.

Curr Genet, 1996 Jul 31, 30(2), 166 - 73
Highly efficient homologous integration via tandem exo-beta-1, 3-glucanase genes in the common mushroom, Agaricus bisporus; van de Rhee MD et al.; Homologous integration was studied in the common mushroom, Agaricus bisporus, using a plasmid (pHAG3-1) carrying the hygromycin-resistance gene and a 3.2-kb genomic fragment from A . bisporus . Homologous integration was found in 30-60% of the transformants obtained with pHAG3-1 linearized at three different positions within the homologous sequence, generating either blunt, 5'- or 3'-protruding ends . The genomic fragment was found to contain two homologous open reading frames in tandem, which showed 60% similarity to exo-beta-1,3-glucanases from Saccharomyces cerevisiae and Candida albicans . The level of the corresponding mRNA is low in the vegetative mycelium and relatively high in fruiting bodies . In the vegetative mycelium of a transformant with tandemly integrated pHAG3-1 plasmids at the homologous position, exoglucanase mRNA was strongly increased without any apparent effect on growth rate or morphology.

J Ethnopharmacol, 1996 Jul 5, 52(3), 171 - 4
Screening of some Cuban medicinal plants for antimicrobial activity; Martinez MJ et al.; The antimicrobial activities of 23 extracts of 12 Cuban plant species reported in traditional medicine were tested . The agar diffusion method was used to assess the activity against four bacteria and one yeast: Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans . The results, evaluated as the diameter of the inhibition zone of microbial growth, showed that nine extracts were active against Gram-positive bacteria but only two of these proved to be also active against Gram-negative bacteria . None of the extracts inhibited the growth of the yeast . The most susceptible bacterium was Staphylococcus aureus and the best antibacterial activity was shown by Schinus terebenthifolius.

J Ethnopharmacol, 1996 Jul 5, 52(3), 151 - 6
Carrier herbal medicine: an evaluation of the antimicrobial and anticancer activity in some frequently used remedies; Ritch-Krc EM et al.; The antimicrobial properties of some traditional Carrier herbal preparations were evaluated using an agar dilution method . Pitch preparations were screened against known human pathogens: Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Aspergillus fumigatus . The results indicated definite antimicrobial activity in the pitch preparations of Picea glauca and Pinus contorta and provide a starting point for pharmacognostic evaluation of these species . In addition, cytoxicity assays, to test the anticancer activity of methanolic extracts of Alnus incana and Shepherdia canadensis against mouse mastocytoma cells, were shown to be positive.

Mycoses, 1996 Jul-Aug, 39(7-8), 271 - 7
The morphology of Candida albicans in two different Earle base media in the presence of tunicamycin; Vespa MN et al.; The effect of tunicamycin on the morphology of Candida albicans yeast cells and germ tubes grown in two different Earle's minimal essential media was investigated . Tunicamycin inhibited germ tube and mycelia formation . Inhibition increased the size and caused aberrant morphology of yeast cells, including bud formation . These cells are hydrophobic and could be used for the production of two monoclonal antibodies suitable for the study of adhesion phenomena as well as ectomural properties.

Acta Otorrinolaringol Esp, 1996 Jul-Aug, 47(4), 325 - 7
{Malignant angular cheilitis}; Seoane J et al.; A case of chronic angular cheilitis is reported . Candida albicans was isolated repeatedly and the process developed into epitheliomatous carcinoma . The etiopathogenic role of Candida albicans and possible mechanism of action are discussed.

J Nat Prod, 1996 Jul, 59(7), 646 - 9
Novel beta-methoxyacrylates of the 9-methoxystrobilurin and oudesmansin classes produced by the basidiomycete Favolaschia pustulosa; Wood KA et al.; Submerged liquid cultures of the basidiomycete Favolaschia pustulosa (Xenova culture collection no . X27732) afforded the novel 9-methoxystrobilurin derivatives, 9-methoxystrobilurin L (1) and 9-methoxystrobilurin E (2), and the related oudemansin derivative, oudemansin L (3) . Their structures were established by 2D NMR experiments . Compounds 1 and 3 possess a novel arrangement of two isoprenoid units fused to the aromatic nucleus . Both 1 and 2 have the EEE-configuration in the pentadienyl side chain as reported previously for 9-methoxystrobilurins . Compound 1 was cytotoxic to cells of the human B lymphoblastoid cell line (Jijoye), with an IC50 of 1.8 nM . This cytotoxicity was observed in a 5- day assay only and was not apparent after 2 days . Compound 1 showed some antibacterial activity against Bacillus subtilis (MIC = 0.9 microM) and antifungal activity against Candida albicans (MIC = 6 microM).

APMIS, 1996 Jul-Aug, 104(7-8), 591 - 7
Preserved neutrophil response to Candida albicans stimulation in AIDS patients with candida esophagitis; Jensen T et al.; Polymorphonuclear neutrophils (PMNs) and their interaction with immunoglobulins constitute a major line of defence against invading Candida albicans . The function of neutrophils, assessed by superoxide production, and the opsonizing efficacy of sera from 15 AIDS patients with esophageal candidiasis and 15 healthy control subjects were studied . When stimulated with opsonized C . albicans the superoxide generation of PMNs from AIDS patients did not differ from the response observed in healthy subjects . However, a significant depression was demonstrated when PMNs were maximally stimulated by phorbol-12-myristate-13-acetate (PMA) . A reduction in opsonizing capability of serum from AIDS patients was detected when tested with zymosan particles . However, the opsonizing capacities of serum from AIDS patients and control subjects were comparable in anticandidal activity, a result that may be explained by a compensatory stimulation of the specific humoral anticandidal response due to perpetual mucous candidiasis in the AIDS patients . These results suggest that anticandidal activity of PMNs and sera from AIDS patients with esophageal candidiasis is preserved, matching the clinical evidence that systemic candidiasis is seldom seen in non-neutropenic AIDS patients.

Diagn Microbiol Infect Dis, 1996 Jul, 25(3), 117 - 21
Rapid detection of susceptibility to fluconazole in Candida species by a bioluminescence assay of intracellular ATP; Kretschmar M et al.; Infections with Candida albicans and Candida species and antifungal resistance are increasingly recognized . The detection of fluconazole resistance is indispensable . We, therefore, compared two rapid methods that use commercially available test kits with the proposed standard of the NCCLS for fluconazole testing . When strains of Candida albicans and Candida species susceptible or resistant to fluconazole were used, measurement of ATP content was superior to the other test because an excellent correlation was obtained already after 5 hours of incubation . The measurement of the metabolic reduction of XTT yielded comparable results, but 24 hours of incubation were necessary.

J Oral Pathol Med, 1996 Jul, 25(6), 308 - 10
Implantation of Candida albicans and other Candida species in the oral cavity of rats; Totti MA et al.; The carriage of five Candida species in the mouths of normal and sialoadenectomised rats was determined for periods up to 30 days after inoculation into the oral cavity . In both test and control animals, Candida albicans was the species recovered in greatest quantities at all periods, followed by C . parapsilosis and C . tropicalis . In contrast, C . guilliermondii and C . krusei were isolatable only in small numbers and only from the 1st up to the 5th day; they were not present thereafter . Sialoadenectomy favoured oral colonisation only by C . albicans (P < 0.05) and did not influence the carriage of the other species.

J Antimicrob Chemother, 1996 Jul, 38(1), 67 - 73
Spectrophotometric determination of the morphogenetic transformation by synchronous Candida albicans: effects of antifungal agents; Hawser S et al.; A spectrophotometric method for studying the morphogenetic transformation by Candida albicans is described . Synchronous yeast-phase C . albicans cells incubated at 35 degrees C formed germ-tubes and resulted in an absorbance increase at 340 nm but not at 595 nm . Furthermore, measurements of germ tube length increases correlated strongly (r2 > 0.95) with increases in absorbance at 340 nm during the morphogenetic transformation . Amphotericin B significantly affected the morphogenetic transformation, at the MIC and 1/10 MIC, whereby no increase in absorbance was observed at 340 nm . In contrast, identical measurements demonstrated that ketoconazole, at the MIC and 1/10 MIC, did not exhibit the same effects on the morphogenetic transformation by C . albicans cells.

J Infect, 1996 Jul, 33(1), 49 - 51
In situ management and molecular analysis of candidaemia related to a totally implantable vascular access in a cystic fibrosis patient; Bonacorsi SP et al.; We describe a case of candidaemia in a paediatric cystic fibrosis (CF) patient with a totally implantable vascular access (TIVA) . Serial quantitative blood cultures during therapy with amphotericin B delivered via the catheter suggested that the patient was responding to therapy . The TIVA was finally removed because of persistent fever, but its culture remained sterile . Randomly amplified polymorphic DNA (RAPD) analysis of Candida albicans from various anatomical sites showed that the patient's sputum was the most likely source of TIVA contamination . Investigation of TIVA-related candidaemia by molecular analysis could guide rational antifungal chemoprophylaxis of TIVA-related candidaemia.

Arzneimittelforschung, 1996 Jul, 46(7), 711 - 5
Effects of amphotericin B incorporated into liposomes and in lipid suspensions in the treatment of murine candidiasis; Kretschmar M et al.; The effects of similar amounts of amphotericin B (CAS 1397-89-3, AmB) in different preparations either as conventional amphotericin B (des-AmB), or liposomal AmB (lipos-AmB), or des-AmB dissolved in a lipid emulsion (lipid-AmB) on Candida albicans and other Candida species were compared in several in vitro and in vivo models . The minimal inhibitory concentration (MIC) of des-AmB was equal to the MIC of lipid-AmB when determined after 24 h . In contrast, the MIC of lipos-AmB was 4-8 times the MIC of des-AmB . When tested at 4 times the MIC of the respective preparations suspension of lipid-AmB led to a reduced ability to kill the fungi whereas des-AmB reduced the inoculum by 99% within 6 h . Four times the MIC of lipos-AmB failed completely to kill the fungi in the same time, but was only fungistatic . At 24 h all preparations had killed the yeasts at concentrations 4 times the MIC . In contrast to the in vitro data, lipos-AmB was more active in the treatment of murine candidiasis than lipid-AmB and des-AmB . Lipos-AmB but not lipid-AmB or des-AmB was able to significantly reduce the amount of Candida albicans in the liver when given in the same dosage . Concomitantly, AmB measured by HPLC was highly concentrated in the livers of the mice treated with lipos-AmB . It is concluded that even when given in the same dosage as des-AmB and lipid-AmB, lipos-AmB is more effective in the treatment of murine candidiasis, although it is less effective in vitro . Lipid-AmB is no alternative to lipos-AmB in this model of systemic infection of mice with Candida albicans.

Yeast, 1996 Jul, 12(9), 893 - 8
A Candida albicans gene encoding a DNA ligase; Andaluz E et al.; A DNA ligase-encoding gene (Ca CDC9) was cloned from Candida albicans by complementation of an ime-1 mutation in Saccharomyces cerevisiae . In this system, IME1 function was assayed using a S . cerevisiae strain with a ime2-promoter-lacZ gene fusion such that following transformation with a C . albicans genomic library, the presence of positive clones was indicated upon the addition of X-gal to sporulation media . Transforming fragments were subcloned in pGEM7 and sequenced . Sequence homology with several ATP-dependent DNA ligases from viruses, fission yeast, human, baker yeast and bacteria was observed.

Ann Plast Surg, 1996 Jul, 37(1), 91 - 3
Candida albicans infection of bilateral polyurethane-coated silicone gel breast implants; Niazi ZB et al.; We present a report of bilateral Candida albicans infection of polyurethane-coated silicone gel prostheses and an acute onset of unilateral capsular contracture 4 years after breast augmentation . The patient was treated by removal of implants, antibiotic irrigation of the capsule cavities, and immediate replacement with new implants . Following histopathologic diagnosis, the patient was treated with a course of fluconazole and remains symptom free at the 12-month follow-up.

Ann Pharmacother, 1996 Jul-Aug, 30(7-8), 765 - 7
Amphotericin B-associated hypertension; Le Y et al.; OBJECTIVE: To report two cases of hypertension related to amphotericin B infusion . CASE SUMMARY: A 63-year-old woman with Candida albicans bacteremia and an 84-year-old man with Aspergillus fumigatus pneumonia developed hypertension within minutes of amphotericin B administration . Both patients' blood pressure returned to baseline soon after the infusion of amphotericin B was stopped . Neither patient was rechallenged . DISCUSSION: A causal relationship may exist between the administration of amphotericin B and these hypertensive episodes . Blood pressure in both patients normalized without treatment on discontinuation of the infusion . The mechanism of amphotericin B-associated hypertension is unclear but could include vasoconstricting properties of the drug or the administration of intravenous NaCl 0.9% prior to amphotericin B infusion . We recommend that intravenous NaCl 0.9% be administered following amphotericin B infusion and that the infusion be stopped if hypertensive episodes arise . CONCLUSIONS: Both acute hypertension and hypotension can occur in patients receiving amphotericin B for systemic fungal infections.

Clin Infect Dis, 1996 Jul, 23(1), 176 - 8
Spondylodiskitis due to Candida albicans: report of two patients who were successfully treated with fluconazole and review of the literature; Hennequin C et al.; We report the cases of two patients with spondylodiskitis due to Candida albicans who were successfully treated with fluconazole . On the basis of findings from these cases and a review of 52 mycologically proven cases in the literature, we describe the main characteristics of candidal spondylodiskitis . In 60% of the cases, candidal spondylodiskitis was a late complication of candidemia (mean delay, 5.2 months) it was determined to be a complication on the basis of the results of previously positive blood cultures (19 cases), and it was presumed to be a complication in iv drug addicts (12 cases) . As spondylodiskitis can be a late complication of candidemia, all episodes of candidemia should be treated with systemic antifungal agents . Clinical and radiological signs of candidal spondylodiskitis were nonspecific . Any bone or joint symptoms in a patient who has had candidemia should be considered to be of fungal origin at the time of presentation . The definitive diagnosis of candidal spondylodiskitis was made on the basis of the results of percutaneous puncture in 26 of 30 cases . The overall prognosis for patients with candidal spondylodiskitis was good, with the full recovery rate ranging from 67% to 100% . The preliminary results of treating candidal spondylodiskitis with triazole derivatives, particularly fluconazole, were satisfactory; there was excellent tolerance of this drug.

Clin Microbiol Rev, 1996 Jul, 9(3), 335 - 48
Immunopathogenesis of recurrent vulvovaginal candidiasis; Fidel PL Jr et al.; Recurrent vulvovaginal candidiasis (RVVC) is a prevalent opportunistic mucosal infection, caused predominantly by Candida albicans, which affects a significant number of otherwise healthy women of childbearing age . Since there are no known exogenous predisposing factors to explain the incidence of symptomatic vaginitis in most women with idiopathic RVVC, it has been postulated that these particular women suffer from an immunological abnormality that prediposes them to RVVC . Because of the increased incidence of mucosal candidiasis in individuals with depressed cell-mediated immunity (CMI), defects in CMI are viewed as a possible explanation for RVVC . In this review, we attempt to place into perspective the accumulated information regarding the immunopathogenesis of RVVC, as well as to provide new immunological perspectives and hypotheses regarding potential immunological deficiencies that may predispose to RVVC and potentially other mucosal infections by the same organism . The results of both clinical studies and studies in an animal model of experimental vaginitis suggest that systemic CMI may not be the predominant host defense mechanism against C . albicans vaginal infections . Rather, locally acquired mucosal immunity, distinct from that in the peripheral circulation, is now under consideration as an important host defense at the vaginal mucosa, as well as the notion that changes in local CMI mechanism(s) may predispose to RVVC.

Chemotherapy, 1996 Jul-Aug, 42(4), 273 - 9
Influence of human serum on the postantifungal effect of four antifungal agents on Candida albicans; Minguez F et al.; This study evaluates the influence of both fresh and heated human serum on the postantifungal effect (PAFE) induced by different concentrations of amphotericin B (AmB), 5-fluorocytosine (5-Fc), Ketoconazole (Kz) and fluconazole (Flu) on two strains of Candida albicans . The concentrations were selected in harmony with the pharmacokinetic properties and toxicity of the drugs . Without serum there was no delay in the growth of yeast cultures pretreated with Kz or Flu, leading to negative PAFEs, however with AmB and 5-Fc the PAFEs were positive . When assays were made in the presence of 10% fresh human serum, the duration of the PAFEs increased with all drugs tested, and those induced by azolic agents became positive . In the presence of 10% human serum heated at 56 degrees C for 30 min, the PAFEs of the antifungal agents were similar to those obtained in the absence of serum . Our results suggest that fresh serum positively influenced PAFE which may be an important factor in determining the dosing regimen for infection by yeasts.

J Clin Microbiol, 1996 Jul, 34(7), 1813 - 4
A single strain of Candida albicans associated with separate episodes of fungemia and meningitis; Porter SD et al.; Four isolates of Candida albicans recovered from the blood and cerebral spinal fluid of a 66-year-old man during episodes of systemic infection separated by 3 months and antifungal therapy were analyzed by a variety of molecular typing methods . All four isolates were shown to represent the same strain, indicating a relapse of infection rather than reinfection.

J Clin Microbiol, 1996 Jul, 34(7), 1677 - 81
DNA fingerprinting of Candida rugosa via repetitive sequence-based PCR; Redkar RJ et al.; A repetitive sequence-based PCR (rep-PCR) technique was developed to characterize the genotypic relatedness among Candida rugosa isolates . Two repetitive sequences, viz., Care-2 and Com29 from Candida albicans, were used to design primers Ca-21, Ca-22, and Com-21, respectively . When used alone or in combination, these primers generated discriminatory fingerprints by amplifying the adjacent variable regions of the genome . Twenty-three isolates from burn patients, eight from other human sources, and four C . rugosa isolates pathogenic in animals were placed into nine fingerprinting groups . Different primers placed these isolates into identical groups, indicating that rep-PCR is a specific and reproducible technique for molecular characterization of C . rugosa . Moreover, these primers unequivocally discriminated among other important Candida species such as C . albicans, C . glabrata, C . tropicalis, C . krusei, C . parapsilosis, C . kefyr, and C . lusitaniae . These data confirm the conservation of repetitive sequences in Candida species . Because of its ease and sensitivity, rep-PCR offers a relatively rapid and discriminatory method for molecular typing of C . rugosa in outbreaks.

Can J Microbiol, 1996 Jul, 42(7), 705 - 710
Effects of amphotericin B on glucose metabolism in Candida albicans blastospores evidenced by 13C NMR; Rabaste F et al.; Candida albicans blastospores harvested from 8- (exponential phase) or 48-h (stationary phase) cultures were incubated with 60 x 10(-3)M {1-(13)C}glucose with or without 10(-4)M amphotericin B (AmB) . The utilization of {1-(13)C}glucose was monitored by in vivo 13C NMR under anaerobiosis . With exponential phase cells, in the presence of AmB, the consumption of glucose and the production of ethanol, trehalose, and glycerol continuously decreased with time, and after 25 min, the metabolism was blocked . On stationary phase cells AmB had almost no effect on glucose metabolism . Comparison with previous experiments evidenced that AmB induced first K+ leakage, then acidification, and finally a stop of the metabolism . In parallel, in vitro 13C NMR spectra were performed on supernatants and cell-free extracts of yeast suspension incubated under the same conditions . For both exponential and stationary phase cells, AmB induced an increase in the membrane permeability to glycerol; no change was observed for the other metabolites.

J Lab Clin Med, 1996 Jul, 128(1), 103 - 14
Soluble (1-->3)-beta-D-glucan purified from Candida albicans: biologic effects and distribution in blood and organs in rabbits; Yoshida M et al.; (1-->3)-beta-D-glucan is a ubiquitous constituent of fungi, and elevated plasma glucan levels are commonly present in patients with deep mycosis or fungemia . The pharmacokinetics, biologic effects, and distribution in blood and organs of iodine 125-labeled (1--> 3)-beta-D-glucan purified from Candida albicans organisms were analyzed in rabbits during the 24-hour period after intravenous administration of this constituent . The intravascular half-life of beta-glucan was 1.8 minutes in the low-dose group (9.3 micrograms/kg, n = 3) and 1.4 minutes in the high-dose group (222 micrograms/kg, n = 3), and the total body clearance was 1.12 +/- 0.30 ml/min and 1.17 +/- 0.16 ml/min (mean +/- SD), respectively (not significantly different) . The serum concentration of (1-->3)-beta-D-glucan was also biologically determined by a test using coagulation factor G of the Japanese horse-shoe crab (G test) . There was good correlation between the clearance of beta-glucan measured biologically and isotopically . During the 24-hour period of observation the rabbits remained well and beta-glucan failed to alter blood cell counts, tumor necrosis factor levels, or lipid metabolism . 125I-labeled beta-glucan associated with the blood cellular compartment initially was less than 3% (the majority in the platelets) and decreased further during the following 2-hour period . Over 97% of circulating 125I-labeled beta-glucan was associated with the cell-free plasma, and the majority of this glucan in plasma appeared not to be associated with lipoproteins . The liver contained more than 80% of the 125I-labeled beta-glucan detected in the six major organs analyzed.

J Leukoc Biol, 1996 Jul, 60(1), 81 - 7
Beta-1,2-linked oligomannosides inhibit Candida albicans binding to murine macrophage; Fradin C et al.; Interaction of Candida albicans with cells of the macrophage lineage was examined by using heat-killed (HK) and live yeast cells . Laminarin, an analogue of the cell wall beta-glucans, strongly inhibited HK yeasts adherence to J774 cell line but had no effect on live yeast binding . Phosphopeptidomannan (PPM) from Saccharomyces cerevisiae had a limited effect on the binding of both HK and live yeasts but significant inhibition was achieved by the use of C . albicans PPM . The role of beta-1,2-oligomannosides was examined with regard to their exclusive presence within C . albicans PPM . PPM acid labile beta-1,2-oligomannosides or a synthetic beta-1,2-mannotetraose, inhibited yeasts binding in a manner comparable to the original PPM . These latter results were confirmed by using mouse peritoneal macrophages, thus suggesting a general role for beta-1,2-oligomannosides in the adherence of the yeast to the macrophage membrane.

Infect Immun, 1996 Jul, 64(7), 2577 - 84
Purification and biochemical characterization of a 65-kilodalton mannoprotein (MP65), a main target of anti-Candida cell-mediated immune responses in humans; Gomez MJ et al.; A 65 kDa-constituent (MP65) of a whole-cell mannoprotein (MP) fraction of Candida albicans was purified by immunoaffinity chromatography with monoclonal antibodies directed against periodate-insensitive, protease-sensitive MP epitopes, putatively polypeptide in nature . These antibodies were obtained by immunization of mice with concanavalin A bead-coupled, low-glycosylated MP from hyphal cells of C . albicans grown in the presence of a subinhibitory dose of tunicamycin . The immunoaffinity-purified MP65 molecule had a pI of 4.1 and a protein/polysaccharide ratio of 1.8:1 . It was resistant to hydrolysis by endoglycosidase H, endoglycosidase F, or N-glycoffanases but still reactive with concanavalin A . The polysaccharide moiety of MP65 was composed exclusively of mannose and glucose at a ratio of 12.7 to 1 . The protein moiety showed numerous potential O-glycosidic linkage sites as suggested by the high proportion of serine and threonine (together accounting for more than 20% of the total amino acid composition) and susceptibility to diluted alkali . This treatment and digestion with alpha-mannosidase caused a reduction in the MP65 molecular mass to around 54 kDa . The N-terminal sequence of MP65 protein moiety was rich in alanine and valine (7 of 13 amino acids) and did not show any significant homology with deposited sequences in data banks . Purified MP65, at doses of a few nanograms, induced extensive T-cell proliferation of human peripheral blood mononuclear cells . This proliferation was specifically inhibited, in a dose-response fashion, by the antigen-binding fragment of the monoclonal antibody used for immunoaffinity purification . Overall, these results highlight biochemical and molecular details of MP65, a main target of human T-cell response to C.albicans.

J Infect Dis, 1996 Jul, 174(1), 133 - 40
Oral immunization of mice against candidiasis; Jensen J et al.; Oral immunization with live Candida albicans evoked antibody- and cell-mediated immune responses in gnotobiotic C.B-17 and BALB/c mice . No deaths or systemic candidiasis of endogenous origin were evident in these C . albicans-colonized (pure culture) mice . Histologic examination showed minimal to no infection of the stomach, esophagus, or tongue by C . albicans . Not only were orally immunized mice more resistant to systemic candidiasis (intravenous challenge) than were germfree (nonimmunized) controls, but immunity could be transferred to susceptible mice with immune spleen cells . Oral immunization elicited a Th1-type response in spleen cells and a Th2 response in Peyer's patch lymphocytes . The alimentary tracts of these orally immunized mice remained chronically colonized with C . albicans in spite of the presence of both antibody- and cell-mediated immune responses to C . albicans.

Yeast, 1996 Jun 30, 12(8), 741 - 56
The Candida albicans PKC1 gene encodes a protein kinase C homolog necessary for cellular integrity but not dimorphism; Paravicini G et al.; Using a DNA fragment derived from the Saccharomyces cerevisiae protein kinase C gene (PKC1) as a probe to screen an ordered array library of genomic DNA from the dimorphic pathogenic fungus Candida albicans, the C . albicans PKC1 gene (CaPKC1) was isolated . The CaPKC1 gene is predicted to encode a protein of 1079 amino acids with 51% sequence identity over the entire length with the S . cerevisiae Pkc1 protein and is capable of functionally complementing the growth defects of a S . cerevisiae pkc1 delta mutant strain on hypo-osmotic medium . Deletion of both endogenous copies of the CaPKC1 gene in diploid C . albicans cells resulted in an osmotically remedial cell lysis defect of both the budding and the hyphal growth form and morphologically aberrant cells of the budding form . Despite these abnormalities, the transition between the two growth forms of C . albicans occurred normally in pkc1/pkc1 double disruptants . Capkc1p was modified at its C-terminus with two repeats of the Staphylococcus aureus protein A IgG-binding fragment (ZZ-sequence tag) and partially purified by chromatography on DEAE-Sepharose and IgG-Sepharose . In vitro, Capkc1p preferably phosphorylated the S . cerevisiae Pkc1p pseudosubstrate peptide and myelin basic protein, but not histones, protamine or dephosphorylated casein, and failed to respond to cofactors known to activate several mammalian PKC isozymes.

Mol Gen Genet, 1996 Jun 24, 251(4), 442 - 50
The Aspergillus nidulans genes chsA and chsD encode chitin synthases which have redundant functions in conidia formation; Motoyama T et al.; We previously isolated three chitin synthase genes (chsA, chsB, and chsC) from Aspergillus nidulans . In the present work, we describe the isolation and characterization of another chitin synthase gene, named chsD, from A . nidulans . Its deduced amino acid sequence shows 56.7% and 55.9% amino acid identity, respectively, with Cal1 of Saccharomyces cerevisiae and Chs3 of Candida albicans . Disruption of chsD caused no defect in cell growth or morphology during the asexual cycle and caused no decrease in chitin content in hyphae . However, double disruption of chsA and chsD caused a remarkable decrease in the efficiency of conidia formation, while double disruption of chsC and chsD caused no defect . Thus it appears that chsA and chsD serve redundant functions in conidia formation.

Int J Paediatr Dent, 1996 Jun, 6(2), 95 - 100
Oral carriage of Candida species in children and adolescents with Down's syndrome; Carlstedt K et al.; Oral carriage of Candida albicans was studied in 55 children and adolescents with Down's syndrome (DS), aged between 7 months and 20 years 6 months, and compared with an age- and sex-matched control group of subjects . Twenty-two of the DS subjects were diagnosed as having congenital cardiovascular malformations . Compared to controls, the DS subjects were more prone to infections . The number of subjects colonized with C . albicans in the oral cavity was significantly higher in the DS group (69%) than in the control group (35%) . Colonization with C . albicans and simultaneous erythematous or white pseudomembranous lesions of the oral mucosa were diagnosed in 22 (40%) of the DS groups and in only one of the control group . In both the DS and the healthy control subjects the frequency of colonization with C . albicans was positively correlated to age . The DS subjects were significantly more densely colonized by C . albicans than the controls . Abnormalities of the immune response in DS children may contribute to the increased oral carriage of C . albicans.

Arch Oral Biol, 1996 Jun, 41(6), 517 - 22
Phospholipid profiles in the oral yeast Candida; Abdi M et al.; Individual molecular species of phospholipids can now be readily detected using fast atom-bombardment mass spectrometry (FAB MS) . The aim was to obtain detailed information on individual molecular species of Candida phospholipids using FAB MS . Lipids were extracted from Candida isolates and prepared for FAB MS analysis in negative-ion mode . Each isolate yielded several hundred anion peaks . The major anion peaks consistent with the presence of phospholipids included those of mass/charge (m/z) 836, 743, 715, 744 and 834 tentatively identified as phosphatidylethanolamine PE(43:5), phosphatidylglycerol PG(34:3), PG(32:3), PE(36:1) and PE(43:5) . Major peaks consistent with the presence of carboxylate anions were of m/z 241, 253, 255, 277, 279, 281, 283, and 307 putatively identified as C15:0, C16:1, C18:3, C18:2, C18:1, C18:0 and C20:2 which also support the putative identifications of phospholipids . Such phospholipid profiles differ from those published for other microorganisms analysed by FAB MS . Quantitative differences were observed between different candidal species . Candida albicans may be readily differentiated from C . glabrata on the basis of its carboxylate anions . Within species, there are quantitative differences in phospholipid profiles . Thus, Candida has a unique combination of phospholipid analogues with potential chemosystematic significance.

J Diarrhoeal Dis Res, 1996 Jun, 14(2), 110 - 2
Diarrhoea associated with Candida spp.: incidence and seasonal variation; Chaudhury A et al.; To study the incidence and seasonal variation of diarrhoea associated with Candida, 978 diarrhoeal stool specimens from patients of all age groups were examined by microscopy and culture . Candida spp . was the sole pathogen (unassociated with other diarrhoeagenic bacteria, protozoa, or helminths) in 15.3% of the total cases . Candida albicans (94.9%) was the predominant species isolated . The incidence was highest among the infants aged 0-12 months (37.1%), followed by a decline in the rest of the children aged less than 5 years with a second peak in the people aged over 5 years, including adults . The paediatric age group had a significantly higher incidence in the summer season compared to the rainy (p < 0.05) or winter (p < 0.01) months . Thus, there was a definite age and seasonal variation in the incidence of diarrhoea caused by the overgrowth of Candida in the Varanasi region of the Indian subcontinent.

Eur J Clin Microbiol Infect Dis, 1996 Jun, 15(6), 503 - 6
Candida endophthalmitis in non-neutropenic critically ill patients; Nolla-Salas J et al.; Six non-neutropenic critically ill patients who developed hematogenous endophthalmitis due to Candida spp . were studied prospectively . In all cases the yeast was isolated in blood cultures . The incidence of endophthalmitis in patients with candidemia was 13%, the predominant species being Candida albicans . Four patients were treated with fluconazole, but its efficacy could not be evaluated because three of the patients died . In patients at risk of candidemia, regular ophthalmoscopic examinations are recommended in order to enable early initiation of systemic antifungal therapy in those who develop endophthalmitis.

Cell Mol Biol (Noisy-le-grand), 1996 Jun, 42(4), 567 - 76
Reassessment of the effect of glucagon and nucleotides on Candida albicans germ tube formation; Zelada A et al.; The role of cyclic AMP in the process of germ tube formation in Candida albicans was investigated . The exogenous supply of the nucleotide or of agents that raise its intracellular levels stimulated germination induced by N-acetyl-D-glucosamine; glucagon showed this same stimulatory effect on yeast cell transition to the hyphal form . Compounds, included glucagon, that stimulated hyphal formation, also notably enhanced the development of hyphae . The stimulatory effect of glucagon on germination was blocked by the specific antagonist des His1 {Glu9} glucagon amide, probably indicating an interaction of the hormone with a glucagon-like receptor on the membrane of the cells . Indirect immunofluorescence experiments showed that glucagon binds to the yeast cell surface . When N-acetyl-D-glucosamine was replaced by serum as inducing agent of germination, the stimulatory effect of glucagon was substantially augmented, the resulting of germination being more than 2.5-fold greater than that attained in the presence of N-acetyl-D-glucosamine; moreover, the glucagon concentration needed for half maximal stimulatory activity with serum as inducing agent was at least 50-fold lower than with N-acetyl-D-glucosamine . Monoclonal and polyclonal anti-glucagon antibodies blocked the effect of the hormone . An interesting result observed during these experiments was the fact that a definite period of incubation of C . albicans yeast cells with N-acetyl-D-glucosamine as inducer commits them to hyphal development . When serum was used as inducer, only yeast cells evaginated during the initial incubation period evolved to the hyphal form upon further incubation in the absence of serum.

J Dermatol Sci, 1996 Jun, 12(2), 140 - 6
Candida albicans induces selective expansion of human T lymphocytes expressing the T-cell receptor variable region V beta 5.1; Walsh P et al.; Candida albicans is a common pathogen which can present major problems as an opportunistic skin pathogen in patients with immunodeficiency . The exact nature of the T cell responses to C . albicans is poorly understood . The purpose of this study was to determine whether C . albicans could stimulate the selective expansion of T lymphocytes expressing particular V beta gene segments . Human T lymphocytes stimulated in vitro with an extract of C . albicans were analyzed for T cell receptor V beta gene expression by using a quantitative PCR technique . We found that stimulation of peripheral blood mononuclear cells (PBMC) produced a selective increase in the expression of V beta 5.1 and 5.2 gene transcripts . Using cytofluorographic analysis with available anti-V beta monoclonal antibodies, we verified that there was a significant selective expansion (P = 0.035) of V beta 5.1 positive T lymphocytes in PBMC from six subjects stimulated in vitro with C . albicans . PCR analysis of V beta 5.1 expansion in 10 subjects showed increases in V beta 5.1 gene transcripts in 7/10 subjects . More importantly, analysis of the T cell infiltrate 48 h after intradermal injections with C . albicans also showed significant expression of V beta 5.1 in the infiltrates, along with the infiltration of V beta 8.1 + T cells . The selective expansion of V beta 5.1 bearing T lymphocytes in PBMC stimulated with C . albicans and in skin test reactions to C . albicans suggests that a restricted population of T cells react to C . albicans . Furthermore, our present data raise the provocative possibility that one or more antigens in C . albicans can act as a superantigen, producing selective expansion of a population of T lymphocytes bearing a particular V beta specificity.

Fungal Genet Biol, 1996 Jun, 20(2), 153 - 67
The chsD and chsE genes of Aspergillus nidulans and their roles in chitin synthesis; Specht CA et al.; Two chitin synthase genes, chsD and chsE, were identified from the filamentous ascomycete Aspergillus nidulans . In a region that is conserved among chitin synthases, the deduced amino acid sequences of chsD and chsE have greater sequence identity to the polypeptides encoded by the Saccharomyces cerevisiae CHS3 gene (also named CSD2, CAL1, DIT101, and KTI1) and the Candida albicans CHSE gene than to other chitin synthases . chsE is more closely related to the CHS3 genes, and this group constitutes the class IV chitin synthases . chsD differs sufficiently from the other classes of fungal chitin synthase genes to constitute a new class, class V . Each of the wild-type A . nidulans genes was replaced by a copy that had a substantial fraction of its coding region replaced by the A . nidulans argB gene . Hyphae from both chsD and chsE disruptants contain about 60-70% of the chitin content of wild-type hyphae . The morphology and development of chsE disruptants are indistinguishable from those of wild type . Nearly all of the conidia of chsD disruption strains swell excessively and lyse when germinated in low osmotic strength medium . Conidia that do not lyse produce hyphae that initially have normal morphology but subsequently lyse at subapical locations and show ballooned walls along their length . The lysis of germinating conidia and hyphae of chsD disruptants is prevented by the presence of osmotic stabilizers in the medium . Conidiophore vesicles from chsD disruption strains frequently swell excessively and lyse, resulting in colonies that show reduced conidiation . These properties indicate that chitin synthesized by the chsD-encoded isozyme contributes to the rigidity of the walls of germinating conidia, of the subapical region of hyphae, and of conidiophore vesicles, but is not necessary for normal morphology of these cells . The phenotypes of chsD and chsE disruptants indicate that the chitin synthesized by each isozyme serves a distinct function . The propensity of a chsD disruptant for osmotically induced lysis was compared to that of strains carrying two other mutations (tsE6 and orlA::trpC) which also result in reduced chitin content vegetative cell lysis . The concentration of osmotic stabilizer necessary to remedy the lysis of strains carrying the three mutations is inversely related to the chitin content of each strain . This finding directly demonstrates the importance of chitin to the integrity of the cell wall and indicates that agents that inhibit the chsD-encoded chitin synthase could be useful anti-Aspergillus drugs.

FEMS Immunol Med Microbiol, 1996 Jun, 14(2-3), 83 - 94
Early inflammatory responses to Candida albicans infection in inbred and complement-deficient mice; Fulurija A et al.; Inflammatory responses that developed in the footpad during the first 48 h after inoculation of Candida albicans were compared in six genetically defined inbred strains of mice . Tissue responses consisted predominantly of accumulations of polymorphonuclear leucocytes, the magnitude of which was significantly less in mice lacking the fifth component of complement (C5) . Despite this, there was no difference between C5-sufficient and C5-deficient mice in the total infectious burden, nor did depletion of complement by treatment with cobra venom factor cause any detectable reduction in the numbers of inflammatory cells in the area of the lesion . Ablation of granulocytes had no significant effect on the fungal burden over the period of the experiment . Immunisation provided some protection against tissue damage, but did not reduce the number of yeasts at the site of infection.

J Interferon Cytokine Res, 1996 Jun, 16(6), 465 - 70
Production of IL-6 and TNF-alpha by the macrophage-like cell line RAW 264.7 after treatment with a cell wall extract of Candida albicans; Vazquez N et al.; Treatment of the cell wall of Candida albicans with ethylenediamine yields an extract that is antigenic for both the humoral and cell-mediated arms of the immune system . This extract has been shown in previous studies by this laboratory and others to possess potent immunomodulatory activity . We report results here that show that treatment of the macrophage-like cell line RAW 264.7 with the ethylenediamine cell wall (EDA-CW) extract results in an increase in the production of both interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) . Our results also show that the EDA-CW extract possess potent costimulatory activity when combined with interferon-gamma (IFN-gamma) . We have found, on the other hand, that EDA-CW extract-treated cells fail to produce elevated levels of IL-1 either alone or in combination with IFN-gamma as a costimulus . Our analysis also shows that the activation of TNF-alpha production by the EDA-CW extract appears to be at the level of transcription, since Northern blot analysis shows that the increase in the level of TNF-alpha mRNA is essentially identical to the rise in TNF-alpha activity released . We suggest that a component of the immunomodulatory activity of the EDA-CW extract is via the activation of macrophage function.

J Med Vet Mycol, 1996 Jun-Jul, 34(3), 219 - 22
Candida albicans stress mannoprotein, SMP200, enhances tumour necrosis factor secretion in the murine macrophage cell line ANA-1; Pitzurra L et al.; Tumour necrosis factor (TNF) secretory activity has been studied in the murine macrophage cell line ANA-1 following in vitro exposure to Candida albicans 200 kDa stress mannoprotein (SMP200) . Treatment of ANA-1 murine macrophages with 200 kDa stress mannoprotein results in increased TNF secretion . The phenomenon is (i) dose- and time-dependent, (ii) abrogated by 200 kDa stress mannoprotein preincubation with a specific monoclonal antibody, and (iii) dependent on intact murine macrophage Ca2+/calmodulin-dependent protein kinase function.

J Med Vet Mycol, 1996 Jun-Jul, 34(3), 205 - 8
Fluconazole therapy of oropharyngeal candidiasis in a patient with multiple endocrine failure does not correlate with Candida albicans susceptibility to fluconazole in vitro; Field EA et al.; We report a case of oropharyngeal and oesophageal candidiasis in a 23-year-old man with endocrinopathy syndrome . Multiple episodes of infection were treated with topical miconazole, oral ketoconazole (200 mg daily) or oral fluconazole (50 mg daily) over a period of 7 years . The final episode failed to respond to ketoconazole (200 mg daily) or fluconazole (200 mg daily), but was treated successfully by increasing the fluconazole dose to 400 mg daily for 6 months . The patient was maintained on fluconazole 200 mg daily without relapse . Serial Candida albicans isolates from the oral cavity were clonally related by RFLP analyses of genomic DNA, and were resistant to fluconazole, ketoconazole and itraconazole in vitro . We conclude that fluconazole 400 mg daily is effective against oropharyngeal and oesophageal candidiasis in a patient with endocrinopathy syndrome, despite the infecting Candida albicans strains being resistant to azole antifungals in vitro.

Trends Microbiol, 1996 Jun, 4(6), 242 - 6
An integrin-like protein in Candida albicans: implications for pathogenesis; Hostetter MK; Candida albicans is the foremost fungal pathogen in neonates, diabetics and immunocompromised people . The discovery of integrin-like proteins in this yeast and the involvement of these proteins in adhesion, morphogenesis and signal transduction suggest important roles for primitive integrins in the pathogenesis of infectious diseases, and as paradigms for signaling and differentiation in vertebrate cells.

Z Gastroenterol, 1996 Jun, 34(6), 361 - 4
Comparison of single-dose and 7-day fluconazole treatment of fungal esophagitis in alcoholic liver disease; Peter Z et al.; The aim of the study was to assess the effectiveness of a single-dose fluconazole treatment of fungal esophagitis in patients with alcoholic liver disease . Twenty-two alcoholic liver disease patients with fungal esophagitis were randomly assigned to receive either a single-dose of 150 mg fluconazole or a 7-day treatment of daily 50 mg fluconazole . Control esophagoscopy was performed in both groups on days 9-11 . Direct smears and cultures on Sabouraud's medium were performed at both endoscopies . Patients' sera were tested for Candida antigens and for antibodies against Candida albicans on days 1, 8 and 15 . Twenty patients (18 C . albicans, 1 C . tropicalis, 1 C . pseudotropicalis) completed the study, there were two drop-outs from the single-dose group . Antibodies against C . albicans were found in four cases, Candida antigens in five . There were no significant differences in the treatment outcome between the two groups, clinical cure was recorded in eight out of nine patients in the single-dose group and nine out of eleven patients in the 7-day group, mycological eradication in four out of nine, and in three out of eleven, respectively . Single-dose fluconazole treatment seems to be an effective therapy of fungal esophagitis in alcoholic liver disease patients.

Br J Dermatol, 1996 Jun, 134 Suppl 46, 22 - 4: discussion 39
Use of terbinafine in HIV-positive subjects: pilot studies in onychomycosis and oral candidiasis; Nandwani R et al.; Study 1 . Eighteen HIV-positive Caucasian homosexual men with initial positive fungal microscopy were recruited into this prospective, dual-centre, open-label study . They received a once-daily oral dose of 250 mg terbinafine for 12 weeks . Eight were subsequently excluded after screening cultures proved negative . The mean CD4 count of the 10 evaluable subjects was 302/mm3 . All 10 positive fungal cultures were confirmed as Trichophyton rubrum . Using an intention-to-treat analysis, healthy unaffected nail growth increased from a mean of 1.6 mm at baseline to 5.2 mm after 12 weeks' treatment . Clinical response after treatment was 6.4 mm at 36 weeks and 8.0 mm at 48 weeks . Three of the 10 toenail infections were cured mycologically . This 30% cure rate was maintained over 48 weeks' follow-up, despite three patients discontinuing the study . One withdrew following a terbinafine-induced drug rash . Two others stopped treatment during HIV-related illnesses, but without terbinafine side-effects . Study 2 . Ten HIV-positive subjects, nine culture-positive for Candida albicans and one for Candida albicans and Candida glabrata, were recruited into this pilot study . They received 250 mg oral terbinafine daily for 14 days . Their average CD4 count was 131/mm3 . All patients remained culture-positive throughout the study . Slight improvements in signs and symptoms were seen in one or two patients but this might well have been attributable to improved oral hygiene . Oral terbinafine at this dosage was therefore not thought an effective treatment for this indication in HIV-positive patients . The drug was well tolerated and no serious treatment-related adverse events were reported.

J Clin Microbiol, 1996 Jun, 34(6), 1542 - 5
Genetic dissimilarity of two fluconazole-resistant Candida albicans strains causing meningitis and oral candidiasis in the same AIDS patient; Berenguer J et al.; We describe a patient with AIDS who simultaneously developed Candida meningitis with three positive cerebrospinal fluid cultures and oral candidiasis . This patient also had a history or recurrent episodes of oral candidiasis treated with fluconazole . The patient did not respond to this therapy but was cured with amphotericin B and flucytosine . In vitro susceptibility tests revealed that each infection was caused by fluconazole-resistant Candida albicans isolates . Strain delineation by karyotyping, NotI restriction pattern analysis, hybridization with the specific probe 27A, and PCR fingerprinting with the phage M13 core sequence clearly demonstrated that meningitis and oral thrush were caused by two genetically different isolates.

J Gastroenterol, 1996 Jun, 31(3), 307 - 13
Esophageal Candida infection and adherence mechanisms in the nonimmunocompromised rabbit; Hoshika K et al.; Candida infection of the esophagus has been reported not only in immunocompromised hosts but also in healthy individuals . However, its mechanisms of action in healthy individuals have not been clarified . Our previous study suggested that physical contact was an important factor for the adherence of Candida albicans . The aim of the present study was to test our hypothesis and clarify the adherence mechanisms . Suspensions of Candida albicans cells were given to rabbits in drinking water without the use of immunosuppressive drugs and/or antibiotics, and the esophagus was examined . Candidial lesions were observed in 14 of 15 rabbits given the suspensions held in water with and without 30% sucrose for 13 days . The number of Candida albicans cells adhering to the esophagus per square millimeter by subepithelial cell insertion was significantly larger than that adhering by attachment . These results indicate that adherence of Candida albicans to the esophagus occurs by sustained physical contact alone under a nonimmunosuppressive state, and that subepithelial cell insertion results in greater attachment on adherence . Our findings provide a clue that may help clarify the mechanism of Candida infection in healthy individuals.

Antimicrob Agents Chemother, 1996 Jun, 40(6), 1382 - 6
Comparison of D0870, a new triazole antifungal agent, to fluconazole for inhibition of Candida albicans cytochrome P-450 by using in vitro assays; Venkateswarlu K et al.; D0870 was 12 to 15 times more active than fluconazole in experiments to determine the MIC for growth arrest for two isolates of Candida albicans . A biochemical comparison of in vitro sterol biosynthesis in cell extracts showed only a twofold superiority of D0870 over fluconazole . A large differentiation (10-fold) in 50% saturating concentrations obtained by examining the binding of the azoles to microsomal P-450 was observed in a type II binding spectrophotometric assay, possibly reflecting the differential affinity for more than one P-450 enzyme . Additional mechanisms besides affinity for the target enzyme sterol 14 alpha-demethylase, such as differential intracellular accumulation of drug, may contribute to the differences in antifungal activity.

Antimicrob Agents Chemother, 1996 Jun, 40(6), 1342 - 5
Comparative resistance of Candida albicans clinical isolates to fluconazole and itraconazole in vitro and in vivo in a murine model; Valentin A et al.; Relationships between azole susceptibility and in vivo response to antifungal therapy in a murine model of candidiasis were investigated for Candida albicans isolates sampled from human immunodeficiency virus type 1-positive patients with oropharyngeal candidiasis . The susceptibilities of seven clinical isolates and two reference strains to fluconazole (FCZ) and itraconazole (ITZ) were determined in vitro by the broth microdilution method . Four isolates were resistant to FCZ and ITZ, two were susceptible to both azoles, and three were resistant to FCZ and susceptible to ITZ (dissociated resistance) . CD1 mice were inoculated with each isolate and treated with either FCZ or ITZ (drug regimen, 5 mg/kg of body weight twice daily for 5 days) . Quantitative cultures of kidneys were performed at the end of the treatment . On the other hand, the survival rates of the mice were followed daily . These two parameters were clearly correlated with in vitro susceptibility . Thus, the phenomenon of a dissociation of resistance to FCZ and ITZ may be found in vivo as well as in vitro.

Genetics, 1996 Jun, 143(2), 769 - 76
DLH1 is a functional Candida albicans homologue of the meiosis-specific gene DMC1; Diener AC et al.; DMC1/LIM15 homologue 1 (DLH1), a gene related to meiosis-specific genes, has been isolated from Candida albicans, a fungus thought not to undergo meiosis . The deduced protein sequence of DLH1 contains 74% amino acid identity with Dmc1p from Saccharomyces cerevisiae and 63% with Lim15p from the plant Lilium longiflorum, meiosis-specific homologues of Escherichia coli RecA . Candida DLH1 complements a dmc1/dmc1 null mutant in S . cerevisiae . High copy expression of DLH1 restores both sporulation and meiotic recombination to a Saccharomyces dmc1 delta/dmc1 delta strain . Unlike the DMC1 gene, which is transcribed only in meiotic cells, the heterologous Candida DLH1 gene is transcribed in both vegetative and meiotic cells of S . cerevisiae . Transcription of DLH1 is not detected or induced in C . albicans under conditions that induce DMC1 and meiosis in S . cerevisiae . The presence of an intact homologue of a meiosis-specific gene in C . albicans raises the possibility that this organism has a cryptic meiotic pathway.

Biomaterials, 1996 Jun, 17(11), 1055 - 9
In vitro antimicrobial efficacy of silver iontophoretic catheter; Raad I et al.; A silver iontophoretic catheter was designed consisting of two silver wires connected to an electric power source and disposed in a parallel and helical manner around the proximal subcutaneous segment of a silicone catheter . In an in vitro tunnelled bridge model the silver iontophoretic catheter prevented the migration of Staphylococcus epidermidis from the highly contaminated hub to the sterile tip over a 40-d period . The silver impregnated cuff and electrically charged wires made of aluminium or iron delayed migration for only 72 h . A modified Kirby-Bauer technique, used to test the inhibitory activity of antimicrobial catheters, showed that the silver iontophoretic catheter has a broad spectrum inhibitory activity against Gram-positive bacteria, Gram-negative bacteria and Candida albicans . The silver iontophoretic catheter provides a long-term electrochemical barrier against the migration of organisms from the external contaminated environment into the sterile intravascular compartment.

J Antibiot (Tokyo), 1996 Jun, 49(6), 547 - 52
Ascosteroside, a new antifungal agent from Ascotricha amphitricha . I . Taxonomy, fermentation and biological activities; Gorman JA et al.; Ascosteroside, a novel antifungal compound, was isolated from the culture broth of Ascotricha amphitricha . This compound is an alpha-linked glycoside of a lanostane type triterpenoid . It is active against yeasts such as Candida albicans and Saccharomyces cerevisiae and against filamentous fungi but shows no activity against bacteria . It is not toxic to mammalian cells at concentrations up to 150 microM . In a mouse model, the compound afforded protection comparable to that of ketoconazole.

Am J Ophthalmol, 1996 Jun, 121(6), 643 - 9
Choroidal neovascularization secondary to Candida albicans chorioretinitis; Jampol LM et al.; PURPOSE: To study the clinical histories and courses of six patients with choroidal neovascularization secondary to endogenous Candida albicans chorioretinitis . METHODS: The medical records, fundus photographs, and fluorescein angiograms of six patients who developed C . albicans chorioretinitis secondary to candidemia and who subsequently developed choroidal neovascularization in one or both eyes were reviewed . RESULTS: The six patients ranged in age from 18 to 79 years . Four were women and two men; all but one showed evidence of bilateral chorioretinal scarring secondary to C . albicans chorioretinitis . All patients had been treated successfully with systemic antifungal therapy (amphotericin B) . Two weeks to two years after the chorioretinitis, choroidal neovascularization developed in one eye (four cases) or both eyes (two cases) . The neovascularization on initial examination was subfoveal in four eyes, extrafoveal in three eyes, and juxtafoveal in one eye . Laser photocoagulation was used in four of the eight involved eyes . In these cases, the active choroidal neovascularization was brought under control . In one eye, the patient had submacular surgery for excision of the choroidal neovascular membrane . Final visual acuities ranged from 20/20 to 20/200 in treated eyes and from 20/50 to 20/400 in untreated eyes . CONCLUSION: Choroidal neovascularization is a potential cause of late visual loss in patients who have had C . albicans sepsis and endogenous C . albicans chorioretinitis . Eyes that have chorioretinal scarring from C . albicans chorioretinitis should be watched for the development of choroidal neovascularization . Laser photocoagulation or perhaps surgical excision of the neovascular complex may be of benefit in selected cases.

Cell Immunol, 1996 May 25, 170(1), 91 - 100
Lymphocytes utilize CD11b/CD18 for adhesion to Candida albicans; Forsyth CB et al.; Large granular lymphocytes require adherence to hyphae of Candida albicans to inhibit growth of this fungus . This study was undertaken to identify the lymphocyte surface structures that mediate this adhesion . Monoclonal antibodies specific for epitopes of the alpha subunit (CD11b) and the beta 2 subunit (CD18) of Mac-1 eliminated lymphocyte adhesion to C . albicans hyphae . Significant inhibition of lymphocyte adhesion to C . albicans was also achieved with known protein ligands of Mac-1 . These proteins included the extracellular matrix proteins vitronectin, laminin, and fibrinogen as well as two engineered peptides containing RGD (arginine-glycine-aspartic acid) sequences . Carbohydrates including N-acetyl-D-glucosamine which have been demonstrated to inhibit Mac-l-mediated adhesion to whole yeast and yeast zymosan also blocked lymphocyte adhesion to hyphae . These results identify Mac-1 (CD11b/CD18) as the surface structure that mediates lymphocyte adhesion to C . albicans . A model is proposed for lymphocyte Mac-1 activation by microbial ligands.

J Biol Chem, 1996 May 24, 271(21), 12445 - 50
The mechanism of the acyl-carbon bond cleavage reaction catalyzed by recombinant sterol 14 alpha-demethylase of Candida albicans (other names are: lanosterol 14 alpha-demethylase, P-45014DM, and CYP51); Shyadehi AZ et al.; The Candida albicans sterol 14 alpha-demethylase gene (P-45014DM, CYP51) was transferred to the yeast plasmid YEp51 placing it under the control of the GAL10 promoter . The resulting construct (YEp51:CYP51) when transformed into the yeast strain GRF18 gave a clone producing 1.5 mu mol of P-450/liter of culture, the microsomal fraction of which contained up to 2.5 nmol of P-450/mg of protein . Two oxygenated precursors for the 14 alpha-demethylase, 3 beta-hydroxylanost-7-en-32-al and 3 beta-hydroxylanost-7-en-32-ol, variously labeled with 2H and 18O at C-32 were synthesized . In this study the conversion of {32-2H,32-16O}- and {32-2H,32-18O}3 beta-hydroxylanost-7-en-32-al with the recombinant 14 alpha-demethylase was performed under 16O2 or 18O2 and the released formic acid analyzed by mass spectrometry . The results showed that in the acyl-carbon bond cleavage step (i.e . the deformylation process) the original carbonyl oxygen at C-32 of the precursor is retained in formic acid and the second oxygen of formate is derived from molecular oxygen; precisely the same scenario that has previously been observed for the acyl-carbon cleavage steps catalyzed by aromatase (P-450arom) and 17 alpha-hydroxylase-17,20-lyase (P-45017 alpha,CYP17) . In the light of these results the mechanism of the acyl-carbon bond cleavage step catalyzed by the 14 alpha-demethylase is considered.

Spec Care Dentist, 1996 May-Jun, 16(3), 128 - 33
Salivary secretion and oral health in narcolepsy: a pilot study; Nordgarden H et al.; Complaints of dry mouth and poor dental health are common in persons with narcolepsy . The aim of this study was to investigate whether salivary secretion is reduced in narcolepsy . Persons using tricyclic anti-depressants (TCAs) were excluded, since TCAs are known to reduce salivary secretion . Thus, two patient subgroups were studied, one on central stimulant (CS) treatment (medicated group, n = 12), and one unmedicated group (n = 8), representing all persons with narcolepsy living in the Oslo area meeting these criteria . The survey group and 20 age- and sex-matched healthy control persons without symptoms of dry mouth were examined with respect to the following parameters: unstimulated (UWS) and chewing-stimulated (SWS) whole salivary flow rates, citric-acid-stimulated parotid and submandibular flow rates, buffering effect, and number of some aciduric micro-organisms in the oral cavity . As a group, persons with narcolepsy had lower whole salivary flow rates, a lower buffering effect, and higher Candida albicans scores than the control group . When the patients were divided into the medicated and unmedicated groups, these differences were valid only for the medicated group . Whether the observed differences were effects of CS medication or reflected that these persons were more seriously affected by the disease has to be further explored.

FEMS Microbiol Lett, 1996 May 1, 138(2-3), 113 - 21
Identification of the gene encoding DNA topoisomerase I from Candida albicans; Taylor A et al.; A gene encoding a type I topoisomerase (TOP1) was isolated from Candida albicans, sequenced, and expressed in Saccharomyces cerevisiae . The TOP1 gene was identified from a C . albicans genomic library by hybridization with the product of a polymerase chain reaction with degenerate primer sets encoding regions conserved in other TOP1 genes . A clone containing an open reading frame of 2463 bp and predicted to encode a protein of 778 amino acids with sequence similarity to eukaryotic type I topoisomerases was identified . The C . albicans TOP1 gene restored camptothecin sensitivity and increased the topoisomerase activity in S . cerevisiae, indicating that the DNA fragment encodes a functional C . albicans topoisomerase I.

FEMS Microbiol Lett, 1996 May 1, 138(2-3), 105 - 11
Comparison of responses of DNA topoisomerase I from Candida albicans and human cells to four new agents which stimulate topoisomerase-dependent DNA nicking; Fostel J et al.; DNA topoisomerase I is a potential target for therapeutic antifungal agents predicted to have a fungicidal mode of action . This report describes four agents with varying degrees of selectivity for the fungal topoisomerase I compared to the human enzyme: 5-hydroxy-1H-indole-3-acetic acid (5-HIAA), quinizarin, dibenzo-p-dioxin-2-carboxylic acid and 7-amino-4-hydroxy-2-naphthalenesulfonic acid . Taken together with the response of topoisomerase to camptothecin and aminocatechol, these data suggest that there are sufficient structural differences between the topoisomerase I from Candida albicans and human cells to allow selective targeting of the fungal topoisomerase I over its human counterpart.

Mycoses, 1996 May-Jun, 39(5-6), 177 - 83
Immunological investigations in vaginal mycoses; Mendling W et al.; In 42 women with chronically recurrent and 20 women with acute Candida albicans vulvovaginitis, as well as 14 women with Candida glabrata vaginitis, the following investigations were carried out: determination of protein content and secretory immunoglobulin A (sIgA) in the cervicovaginal secretion by a self-modified ELISA technique; determination of immunocells and cellbound IgA in the cervicovaginal secretion by immunofluorescence and nephelometric analysis of IgA in the serum . The results were compared with those of 77 pre-menopausal non-pregnant women with or without intake of anti-ovulants, 17 healthy pregnant women and four hysterectomised pre-menopausal women . Due to inflammation, women with acute and chronically recurrent Candida albicans vulvovaginitis had a higher protein content in the cervicovaginal secretion than healthy women . However, the content of secretory IgA was not increased but even slightly decreased in chronic cases . The number of macrophages and granulocytes in the vaginal content was not increased compared with healthy patients . In only a few cases was IgA detected on yeast cells and in the cervicovaginal secretion by fluorescence microscopy . In chronically-relapsing vaginal candidosis, the frequency of the serotype B of C . albicans was strikingly high . Women with Candida glabrata vaginitis showed lower values of secretory sIgA in the vaginal secretion compared with healthy patients as well as women with vaginitis caused by C . albicans . However, like healthy women, they had normal protein values in the cervicovaginal secretion and also lower values of IgA in the serum compared with women of C . albicans vulvovaginitis patients . Macrophages and granulocytes were demonstrable in the cervicovaginal secretion just as in healthy persons . Women with C . glabrata vaginitis showed a more conspicuous, although not a significantly more frequent, binding of IgA to budding cells demonstrated by fluorescence microscopy than women with C . albicans.

Mycoses, 1996 May-Jun, 39(5-6), 169 - 76
Characterization of specific anti-Candida IgM, IgA and IgE: diagnostic value in deep-seated infections; Aubert D et al.; The proposed serological diagnosis of systemic Candida infections is based on a microplate immunocapture technique detecting IgM, IgA and IgE anti-Candida antibodies . Activity is revealed with a suspension of human erythrocytes sensitized with somatic antigen of Candida albicans, and is quantified on an automated plate reader . The sera were obtained from patients with deep-seated (n = 56) and superficial (n = 193) candidosis . We compared this immunological method with a combination of indirect immunofluorescence and co-immunoelectrodiffusion . The immunocapture method was more sensitive (80.4% vs . 48.2% with indirect immunofluorescence and 58.9% with co-immunoelectrodiffusion), and often provided the diagnosis at an earlier stage, with clear therapeutic advantages . The IgA isotype was a particularly valuable marker of deep-seated Candida infections.

Mycoses, 1996 May-Jun, 39(5-6), 157 - 60
Comparative virulence of Candida albicans strains in CFW1 mice and Sprague-Dawley rats; Schmidt A et al.; To verify host-species specificities of virulence of Candida albicans in experimental systemic mycoses, 10 ATCC strains of Candida albicans, were compared for their virulence in CFW1 mice and Sprague-Dawley rats . Virulence was parallel in mice and rats, four strains were avirulent (ATCC 10231, 18804, 38245, 44831), one strain had an intermediate virulence (ATCC 32354), and five strains (ATCC 10261, 44373, 44505, 62342, 90028) were highly virulent in both host species . Infection doses of 2 x 10(6) CFU per mouse and 5 x 10(6) CFU per rat were comparable with respect to mortality of animals within 10 days; this represents a 4 : 1 ratio on the basis of body weight . In Sprague-Dawley rats haemorrhage occurred in infections with all virulent strains which was not observed in CFW1 mice.

Infection, 1996 May-Jun, 24(3), 263 - 6
Fluconazole in Candida albicans sepsis during pregnancy: case report and review of the literature; Wiesinger EC et al.; Candida sepsis during pregnancy is a rare but life-threatening complication of infection with Candida albicans . In contrast to the situation with other antimicrobial agents, there exists only limited experience with systemic antifungal therapy during pregnancy . A recent report focuses on amphotericin B treatment in systemic fungal infection during pregnancy . The present report discusses a pregnant patient with Candida albicans sepsis and endophthalmitis as well as candida infection of the oral and genital mucous membranes, after hyperalimentation and broad spectrum antibiotic therapy via a central venous catheter . The patient was treated with 10 mg/kg fluconazole from week 16 of gestation for a total duration of 50 days . Adverse effects did not occur and the rest of the pregnancy proceeded favourably for both the mother and the baby.

Pathol Biol (Paris), 1996 May, 44(5), 447 - 51
{Mycological monitoring of Candida albicans infections in various hospital care units . Molecular typing of isolated strains and epidemiological survey}; Arnavielhe S et al.; To evaluate risk factors associated with the nosocomial infection of Candida albicans, a prospective study is conducted twice for three months in three intensive care units . Samples from patients HIV negatives, non neutropenic and non immunodepressive are collected as they came in the unit, on several anatomic sites . Every C . albicans carriers are included in a mycological monitoring . Samples from environmental surfaces, hands and deep pharynx from hospital personnel were also cultured . Strains genetic profile are defined by isoenzyme electrophoresis technique . Thirteen polymorphic loci allowed samples classement into 52 electrophoretic types (ET) . If only one crossed contamination is described, strains regroupment into some ET incites us to extend this study . C . albicans strains from patients closed environment have never been isolated.

J Antimicrob Chemother, 1996 May, 37(5), 943 - 54
Effect of pentoxifylline on the course of systemic Candida albicans infection in mice; Louie A et al.; Pentoxifylline can decrease the production of tumour necrosis factor alpha (TNF alpha) by endotoxin-stimulated macrophages and may improve survival in animals with overwhelming bacterial sepsis . In this study various doses of pentoxifylline were administered to mice with systemic Candida albicans infection to determine its effect on serum TNF alpha levels, organ fungal burden, and host survival . Intraperitoneal injections of pentoxifylline at 20 mg/kg every 8 h did not affect these endpoints . However, fungal counts were significantly higher in kidneys of animals that received 30 and 60 mg/kg of pentoxifylline every 8 h when compared to controls . Injection of 60 mg/kg of pentoxifylline at 8 h intervals also significantly shortened mean survival from 5.8 to 3.8 days (P = 0.01) . Pentoxifylline did not affect peripheral WBC counts, serum TNF alpha and interleukin-6 levels, or the density of neutrophils in tissues . In vitro, pentoxifylline decreased the production of TNF alpha by C . albicans-stimulated macrophages in a dose-dependent manner, but only at concentrations greater than 100 mg/L . In contrast, pentoxifylline suppressed TNF alpha production by endotoxin-stimulated macrophages at concentrations as low as 10 mg/L . Thus, higher doses of pentoxifylline are detrimental in systemic C . albicans infection . However, the detrimental effect is not mediated by alterations in serum TNF alpha or interleukin-6 levels or the aggregation of neutrophils in tissues.

J Antimicrob Chemother, 1996 May, 37(5), 911 - 8
Antimicrobial activities in vitro and in vivo of transition element complexes containing gold(I) and osmium(VI); Elsome AM et al.; Metal compounds have been used as antibacterial agents for centuries . The in-vitro activity of two metal containing complexes, one gold, the other osmium, was investigated using a panel of clinically isolated bacteria and Candida albicans . Twenty strains of each organism were used and MIC and MBC values determined using the agar plate dilution method . Protein binding effects on the activity of the compounds were also investigated using media supplemented with 5% human blood . In-vivo activity of the two compounds was subsequently determined in a hairless-obese mouse skin-surface activity model . Both compounds were highly active against the Gram-positive organisms and Candida albicans in vitro . The gold compound had some Gram-negative activity but the osmium complex was inactive against these organisms . Both were extensively protein bound . In the in-vivo experiment the gold compound achieved a 2-3 log reduction for all the test organisms and was at least as good as or superior to mupirocin in its eradication rate . The osmium compound was inactive.

J Ethnopharmacol, 1996 May, 52(1), 27 - 33
Preliminary antimicrobial screening of four South African Asteraceae species; Salie F et al.; Organic and aqueous solvent extracts of Arctotis auriculata Jacq., Eriocephalus africanus L., Felicia erigeroides DC., and Helichrysum crispum (L.) D . Don, were investigated for selective antimicrobial activities . Organic extracts of A . auriculata and H . crispum inhibited the growth of Mycobacterium smegmatis . The same extracts, together with organic extracts of F . erigeroides, were active against Pseudomonas aeruginosa . Antifungal activities against Candida albicans were exhibited by organic extracts of E . africanus, F . erigeroides, and H . crispum . Organic extracts of A . auriculata and E . africanus, as well as the aqueous extract of the latter plant, were active against Staphyllococcus aureus.

J Clin Microbiol, 1996 May, 34(5), 1235 - 48
Typing Candida albicans oral isolates from human immunodeficiency virus-infected patients by multilocus enzyme electrophoresis and DNA fingerprinting; Boerlin P et al.; A total of 189 Candida albicans isolates have been typed by multilocus enzyme electrophoresis . The results obtained confirm the clonal mode of reproduction of C . albicans . The C . albicans populations found in the oropharynx of human immunodeficiency virus (HIV)-infected patients, in the oropharynx of healthy carriers, or in association with invasive candidiasis could not be distinguished . No clone or group of clones could be associated with the appearance of clinical disorders or with a reduced in vitro susceptibility to the antifungal agent fluconazole . Multiple and sequential oral isolates from 24 HIV-infected patients were also typed by restriction enzyme analysis with the enzymes EcoRI and HinfI and by use of the Ca3 repetitive probe . The results obtained by the combination of all three typing methods show that all but one patient each carried a unique major C . albicans clone in their oropharynx . The 21 patients with sequential isolates had the same C . albicans clones in their throats during recurrent oropharyngeal candidiasis episodes, independently of clinical status or of changes of in vitro susceptibility to fluconazole . Finally, several isolates of the same C . albicans clone found simultaneously in the oropharynx of a patient may present different levels of susceptibility to fluconazole.

AIDS, 1996 May, 10(5), 477 - 83
Inhibition of fungicidal activity of polymorphonuclear leukocytes from HIV-infected patients by interleukin (IL)-4 and IL-10; Tascini C et al.; OBJECTIVE: To investigate the effect of human recombinant interleukin (hrIL)-4 or hrIL-10 on the functional status of polymorphonuclear leukocytes (PMNL) from normal subjects and HIV-infected patients . DESIGN: In an in vitro system we studied the effect of hrIL-4 or hrIL-10 on phagocytosis, fungicidal activity and superoxide anion production by PMNL . METHODS: PMNL were treated in vitro with hrIL-4 or hrIL-10 or their combination for 6 h and then candidacidal activity was evaluated in a colony-forming unit inhibition assay . Superoxide anion generation by PMNL was measured in the presence or absence of preopsonized zymosan or Candida albicans . RESULTS: Treatment in vitro with hrIL-4 or hrIL-10 of PMNL for 6 h was able to impair candidacidal activity of neutrophils in both normal or HIV-infected patients . The inhibitory effect was time- and dose-dependent and was more evident in PMNL from HIV-infected subjects, and reflected in these latter cells a decrease of superoxide anion generation . The impairment of candidacidal activity in PMNL from HIV-infected patients was accompanied by survival of the yeasts shown by budding formation into phagosomic organelles of cytokine-treated PMNL . CONCLUSIONS: Our data highlight new biological effects of IL-4 and IL-10 evidenced by suppressed effector function of neutrophils; this phenomenon is emphasized in HIV-infected patients suggesting a role for these cytokines in mediating increased susceptibility to microbial infection during AIDS progression.

Antimicrob Agents Chemother, 1996 May, 40(5), 1317 - 20
Variation in fluconazole efficacy for Candida albicans strains sequentially isolated from oral cavities of patients with AIDS in an experimental murine candidiasis model; Barchiesi F et al.; Four strains of Candida albicans, isolated from two patients with AIDS who had undergone prolonged fluconazole therapy for oral candidiasis, were studied in a model of disseminated murine candidiasis . Pre- and posttreatment isolates from each patient were genetically related, and the fluconazole MICs for the strains had increased significantly, from 0.25 to 32 micrograms/ml for the strains isolated from patient 1 and from 1.0 to 16 micrograms/ml for the strains isolated from patient 2 . Mice were infected intravenously and were treated orally with fluconazole . For survival studies, mice were treated from day 1 to day 10 postinfection and were observed through day 30 . The fluconazole dosages were as follows: 0.25, 0.5, 1.0, and 5.0 mg/kg of body weight twice a day . For tissue burden studies, two groups of mice (each group received fluconazole at 0.25 or 5.0 mg/kg) were treated from day 1 to day 7 and were sacrificed 1 day later for quantitative tissue cultures of the spleen and both kidneys . For pretreatment isolates from both patients, all fluconazole dosing regimens were effective at prolonging survival compared with the survival of the control groups . For posttreatment isolates, only fluconazole at 5.0 mg/kg was effective at prolonging survival . Both fluconazole dosing regimens used in the tissue burden studies significantly reduced the counts of the pretreatment isolate from patient 1 in the spleen and kidney, while fluconazole at 5.0 mg/kg was effective at reducing the counts of the posttreatment isolate . For both isolates from patient 2, only fluconazole at 5.0 mg/kg was effective at reducing the counts in the spleen and kidney . The study indicates that C . albicans mutation to resistance to fluconazole may play a critical role in fluconazole-refractory oral candidiasis in AIDS patients.

Antimicrob Agents Chemother, 1996 May, 40(5), 1277 - 9
In vitro activities of semisynthetic pneumocandin L-733,560 against fluconazole-resistant and -susceptible Candida albicans isolates; Martinez-Suarez JV et al.; Lipopeptide L-733,560 is a water-soluble derivative of pneumocandin B0 that exhibits enhanced anti-Candida activity . We investigated the in vitro activity of L-733,560 compared with those of amphotericin B, flucytosine, and itraconazole, against fluconazole-resistant (n = 44) and fluconazole-susceptible (n = 46) Candida albicans isolates . Tests were performed with a photometer-read broth microdilution method with RPMI-2% glucose and National Committee for Clinical Laboratory Standards reference strains . Except for those of itraconazole, MICs were not significantly different between the two groups of isolates, as expected for agents with different mechanisms of action . L-733,560 was the most active agent against C.albicans, with MICs for 50 and 90% of the strains tested of 0.01 and 0.06 microgram/ml, respectively.

Clin Infect Dis, 1996 May, 22(5), 803 - 8
Vertical and horizontal transmission of unique Candida species to premature newborns; Waggoner-Fountain LA et al.; The number of nosocomial bloodstream infections due to Candida species in critically ill newborns is increasing . This pathogen may be vertically transmitted from the mother or nosocomially acquired in the nursery . The goal of this study was to identify the route of transmission of unique Candida species and strains from mothers to their preterm offspring . Specimens from mothers for fungal cultures were obtained before delivery, and specimens from infants for sequential fungal cultures were obtained at defined intervals . Candida species were identified by standard methods and were typed by electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG) with pulsed-field gel electrophoresis . Antifungal susceptibility testing was performed on all isolates . Fungal cultures were positive for Candida species in 12 (63%) of 19 mothers' specimens and in seven (33%) of 21 infants' specimens . EK and REAG revealed that both the mother and the infant in three (14%) of 21 mother-infant pairs were colonized with the identical strain of Candida albicans . C . albicans was most commonly transmitted vertically . Candida parapsilosis colonized other infants and could not be accounted for by a maternal reservoir.

Clin Infect Dis, 1996 May, 22 Suppl 2, S89 - 94
Nosocomial candidiasis: emerging species, reservoirs, and modes of transmission; Pfaller MA; During the 1980s, the frequency of nosocomial candidiasis increased dramatically . This trend has continued into the 1990s, and Candida species remain a major cause of nosocomial infections . Although Candida albicans remains the most frequent cause of fungemia and hematogenously disseminated candidiasis, a number of reports have documented infections caused by other Candida species: C . tropicalis, C . glabrata, C . parapsilosis, C . krusei, and C . lusitaniae . Many of these infections arise from an endogenous source, and their frequency is influenced by the patient population, the various treatment regimens, and the antibiotics or other supportive care measures employed at specific institutions . Additional infections may be accounted for by exogenous acquisition via the hands of health care workers, contaminated infusates and biomaterials, and the inanimate environment . Ongoing investigation should help improve our understanding of the epidemiology of candidiasis and facilitate the development of rational preventive measures.

Clin Infect Dis, 1996 May, 22 Suppl 2, S73 - 88
The role of the gastrointestinal tract in hematogenous candidiasis: from the laboratory to the bedside; Cole GT et al.; The gastrointestinal (GI) tract is a frequent source of hematogenous candidiasis in humans . Animal models of GI and hematogenous candidiasis have provided insights into the nature of candidal infection of host mucosal tissue, mechanisms of fungal dissemination to body organs, and features of host response to candidal infections . Biological systems such as these that simulate human candidiasis can be used for testing novel antifungal drugs . We have focused on two murine models of candidiasis with similarities to this fungal disease in humans . The first model simulates a commensal association of Candida albicans with the GI tract of immunocompetent hosts; it has permitted studies of innate and immune cell response to long-term ( > 60 days) infection of the esophageal, gastric, and intestinal mucosa . The second model simulates candidal infection in granulocytopenic patients with invasive candidiasis that originated from sites of colonization in the gut . Both models are well suited for investigating new approaches to prevention and treatment of hematogenous candidiasis . A review of the data on the role of GI candidiasis in hematogenous candidal infections is presented.

Clin Diagn Lab Immunol, 1996 May, 3(3), 331 - 6
Antigenicity of cell wall mannans of Candida albicans NIH B-792 (serotype B) strain cells cultured at high temperature in yeast extract-containing sabouraud liquid medium; Okawa Y et al.; Cultivation of Candida albicans NIH B-792 (serotype B) at high temperature (37 degrees C) for 48 h in yeast extract-containing Sabouraud liquid medium (YSLM) provided the following findings in comparison with the findings obtained after incubation at 27 degrees C . Growth of the blastoconidia of this strain was decreased, with a dry weight of 9%, and the cells were deficient in cytokinesis . The cells did not undergo agglutination with serum factor 5 from a commercially available serum factor kit (Candida Check) . Mannan (B-37-M) obtained from the cells cultured at 37 degrees C had partially lost its reactivity against serum factor 4 and lost most of its reactivity against serum factor 5 in an enzyme-linked immunosorbent assay (ELISA) in contrast to that (B-27-M) at 27 degrees C . Both cells and mannan prepared by cultivation first at 37 degrees C and then at 27 degrees C entirely recovered their reactivities with serum factors 4 and 5 . 1H-nuclear magnetic resonance analysis also revealed that B-37-M had lost a beta-1,2-linked mannopyranose unit and retained a phosphate group . Similar changes were observed in the three other serotype B strains used in the study . The beta-1,2-linked mannooligosaccharides longer than mannotetraose were not included among the products released from B-37-M by mild acid treatment . The results of the inhibition ELISA with a series of beta-1,2-linked mannooligosaccharides from biose to octaose (M2 to M8, respectively) showed that the reactivity against serum factor 4 was inhibited most strongly by the oligosaccharides M4 to M8 and that the reactivity against serum factor 5 was inhibited completely by relatively longer oligosaccharides, M5 to M8, indicating their participation as the antigenic factor 5 epitopes.

Clin Diagn Lab Immunol, 1996 May, 3(3), 290 - 4
Detection of candidal antigens in autoimmune polyglandular syndrome type I; Peterson P et al.; Autoimmune polyglandular syndrome type I (APS I) is associated with chronic mucocutaneous candidiasis . To characterize the antibody responses in this subgroup of Candida albicans infections, we screened a candidal cDNA expression library with patient sera and found four cDNA clones encoding the immunopositive proteins enolase, heat shock protein 90, pyruvate kinase, and alcohol dehydrogenase . The reactivity to these antigens was studied further by immunoprecipitation assays with in vitro-transcribed and -translated proteins . Analysis of sera from 44 APS I patients showed that the highest antibody reactivity was found with enolase (80% of patients reactive), but significant serological responses were also found with heat shock protein 90 (67%), pyruvate kinase (62.5%), and alcohol dehydrogenase (64%) . Overall, 95.5% of patients had detectable antibodies to at least one of these proteins . The cDNAs of enolase and heat shock protein 90 were also expressed in Escherichia coli and studied by immunoblotting . Again, 84% of sera reacted with enolase, whereas 44% of sera reacted with heat shock protein 90 . A good correlation between the two methods was found for both enolase (r = 0.86; n = 58; P < 0.001) and heat shock protein 90 (r = 0.71; n = 56; P < 0.001) . Our results indicate that the four abundant candidal proteins are the major antigens and can be used as accurate markers of candidiasis in APS I patients . The immunoprecipitation assay described here is particularly useful for the rapid analysis of a large number of samples.

Microbiology, 1996 May, 142 ( Pt 5), 1239 - 48
Dynamic expression of cell-surface antigens probed with Candida albicans-specific monoclonal antibodies; Deslauriers N et al.; IgG hybridomas were produced with preferentially reacted with cell-surface antigens of either yeast cells or hyphae of Candida albicans . Four mAbs were used in an immunostaining procedure to follow the expression dynamics of these antigens in media supplemented with glucose or galactose . Yeast cell growth was analysed during the lag phase, the early- and late-exponential phases and the stationary phase, and mycelium formation was analysed between 0.5 and 24 h induction at 37 degrees C . It appears that yeast cell-surface antigens 5C11 and 2E11 are expressed throughout all phases of yeast cell growth as well as on young hyphae after up to 1 h induction . Longer hyphae only faintly react with these two mAbs as they switch to hyphal cell-surface antigens 2G8 and 4E1 after 3 h induction . The reactivity to mAbs 2G8 and 4E1 was induced after a 3 h temperature shift and was confined to the terminal third of growing mycelia . Growth and hyphae induction in galactose prolonged the reactivity of young hyphae with the two anti-yeast-cell mAbs, whereas the expression of surface antigens 2G8 and 2E11 appeared delayed and desynchronized on hyphae . Whereas a similar reactivity was found with ten ATCC strains of C . albicans, four clinical isolates had a unique pattern of reactivity . Immunoblot analyses of DTT extracts of cell-surface constituents indicated that the antigens were proteinaceous in nature and showed that yeast-cell antigens 5C11 and 2E11 are detected in four bands between 68 and 104 kDa, whereas mycelial antigens 4E1 and 2G8 are detected in 117 kDa and 104 kDa bands found in mycelial but not in yeast-cell extracts . Present data support the concept of a dynamic balance in the expression of phase-specific antigens in C . albicans.

Kansenshogaku Zasshi, 1996 May, 70(5), 463 - 9
{Suppression of anti-Candida activity of human neutrophils by glucose and diminishment of the glucose effect by an amino acid mixture}; Tansho T et al.; Effects of a glucose and amino acid mixture prescribed for parenteral alimentation on anti-Candida activity of neutrophils were examined . Neutrophils obtained from peripheral blood of healthy humans inhibited the growth of Candida albicans in vitro . More than 1.0% of glucose inhibited the anti-Candida activity of the neutrophils in a dose-dependent manner . This glucose effect was reduced by the addition of an amino acid mixture clinically prescribed with a carbohydrate solution (PN-twin) in Japan . The amino acid mixture neutralized the suppression of anti-Candida activity of neutrophils by dexamethasone . These results suggest that an amino acid mixture prescribed in an alimentation solution may play a role as a neutralizer of the suppressive action of glucose for anti-Candida activity of neutrophils in a limited area near the top of a catheter in a blood vessel.

Toxicol Appl Pharmacol, 1996 May, 138(1), 176 - 85
Time-dependent changes of inflammatory mediators in the lungs of humans exposed to 0.4 ppm ozone for 2 hr: a comparison of mediators found in bronchoalveolar lavage fluid 1 and 18 hr after exposure; Devlin RB et al.; Acute exposure of humans to ozone results in reversible respiratory function decrements and cellular and biochemical changes leading to the production of substances which can mediate inflammation and acute lung injury . While pulmonary function decrements occur almost immediately after ozone exposure, it is not known how quickly the cellular and biochemical changes indicative of inflammation occur in humans . Increased bronchoalveolar lavage (BAL) fluid levels of neutrophils (PMNs) and prostaglandins (PGE2) have been reported in humans as early as 3 hr and as late as 18 hr after exposure . The purpose of this study was to determine whether a broad range of inflammatory mediators are elevated in BAl fluid within 1 hr of exposure . We exposed eight healthy volunteers twice: once to 0.4 ppm ozone and once to filtered air . Each exposure lasted for 2 hr during which the subjects underwent intermittent heavy exercise (66 liters/min) . BAL was performed 1 hr after the exposure . Ozone induced rapid increases in PMNs, total protein, LDH, alpha-1 antitrypsin, fibronectin, PGE2, thromboxane B2, C3a, tissue factor, and clotting factor VII . In addition, there was a decrease in the recovery of total cells and alveolar macrophages, and decreased ability of alveolar macrophages to phagocytize Candida albicans . A comparison of these changes with changes observed in an earlier study in which subjects underwent BAL 18 hr after an identical exposure regimen indicates that IL-6 and PGE2 levels were higher 1 hr after exposure than 18 hr after exposure, fibronectin and tissue-plasminogen activator levels were higher 18 hr after exposure, and that PMNs, protein, and C3a were present at essentially the same levels at both times . These results indicate that (i) several inflammatory mediators are already elevated 1 hr after exposure; (ii) some mediators achieve their maximal levels in BAL fluid at different times following exposure . These data suggest that the inflammatory response is complex, depending on a cascade of timed events, and that depending on the mediator of interest one must choose an appropriate sampling time.

Can J Microbiol, 1996 May, 42(5), 479 - 86
Anti-adhesin antibodies that recognize a receptor-binding motif (adhesintope) inhibit pilus/fimbrial-mediated adherence of Pseudomonas aeruginosa and Candida albicans to asialo-GM1 receptors and human buccal epithelial cell surface receptors; Lee KK et al.; Pseudomonas aeruginosa and Candida albicans were reported to adhere to the glycosphingolipid asialo-GM1 by means of pili and fimbriae, respectively . These diverse adhesins have been previously reported to have an immunologically conserved antigenic epitope and the role of this cross-reactive epitope in adherence to asialo-GM1 was investigated in this study . Both the unbiotinylated PAK pilus and fimbrial adhesins inhibited biotinylated pili from P . aeruginosa PAK and biotinylated C . albicans fimbriae binding to asialo-GM1 and receptors present on human buccal epithelial cells (BECs), which suggested that the same receptor sites were recognized by the two adhesins . Monoclonal antibodies PK99H and Fm16 raised against the P . aeruginosa PAK pili and C . albicans fimbriae, respectively, recognized a conserved epitope present on the two adhesins . Both Fm16 and PK99H blocked fimbriae binding to asialo-GM1 and BEC receptors and also inhibited P . aeruginosa and C . albicans whole cell binding to BECs . These data suggested that the conserved epitope confers receptor-binding properties to the adhesins, demonstrated that (i) asialo-GM1-like receptors present on epithelial cell surfaces are utilized by the pilus and fimbrial adhesins and (ii) the binding to these glycoreceptors is mediated by a conserved epitope that has receptor-binding properties.

J Infect Dis, 1996 May, 173(5), 1202 - 7
Phagocytosis and intracellular killing of Candida albicans by macrophages exposed to myeloperoxidase; Lefkowitz SS et al.; Candida albicans is an opportunistic pathogen whose resurgence coincides with the rising number of AIDS patients . Neutrophils are known to be involved in the clearance of Candida infections; however, the role of macrophages in host defenses against this organism is not well understood . The present study was undertaken to examine an unrecognized interaction between neutrophils and macrophages resulting in enhanced killing of candidae in vitro . Murine peritoneal macrophages exposed to recombinant myeloperoxidase exhibited enhancement of the respiratory burst, increased phagocytosis, and a dose-dependent increase in intracellular killing of Candida species . Radical scavengers reduced the killing, indicating a role of reactive oxygen intermediates in the candidacidal activity observed . These data suggest that at the site of infection, myeloperoxidase released from neutrophils activates macrophages and induces microbicidal activity.

Infect Immun, 1996 May, 64(5), 1866 - 9
Strain-dependent differences in host response to Candida albicans infection in mice are related to organ susceptibility and infectious load; Ashman RB et al.; After systemic infection with the yeast Candida albicans, inbred mice show substantial differences in mortality, organ colonization, and severity of tissue damage . To examine the relationships between these variables, which are not directly correlated with each other, fungal colonization of the kidneys and brain was enumerated in six inbred strains that exhibit different patterns of tissue damage and mortality . Mice lacking the fifth component of complement (C5) are highly susceptible to lethal challenge, and A/J and DBA/2 mice, both C5 deficient, showed the highest colony counts in the kidneys after challenge with 10(5) blastoconidia . In contrast, colony counts in the brain of all six strains were equivalent at this challenge dose . A/J and DBA/2 mice died after challenge with 3 x 10(5) blastoconidia, but other strains showed an increase in kidney colonization, and strain-dependent differences in clearance from the brain became evident . The data suggest that mortality in A/J and DBA/2 mice is related to an unusual susceptibility of the kidneys to colonization by C . albicans and that there may be tissue-specific differences in host protective mechanisms.

Nat Struct Biol, 1996 May, 3(5), 470 - 9
The x-ray crystal structure of phosphomannose isomerase from Candida albicans at 1.7 angstrom resolution; Cleasby A et al.; Phosphomannose isomerase (PMI) catalyses the reversible isomerization of fructose-6-phosphate (F6P) and mannose-6-phosphate (M6P) . Absence of PMI activity in yeasts causes cell lysis and thus the enzyme is a potential target for inhibition and may be a route to antifungal drugs . The 1.7 A crystal structure of PMI from Candida albicans shows that the enzyme has three distinct domains . The active site lies in the central domain, contains a single essential zinc atom, and forms a deep, open cavity of suitable dimensions to contain M6P or F6P The central domain is flanked by a helical domain on one side and a jelly-roll like domain on the other.

Mol Gen Genet, 1996 Apr 24, 251(1), 75 - 80
Stable transformation and regulated expression of an inducible reporter construct in Candida albicans using restriction enzyme-mediated integration; Brown DH Jr et al.; To allow the regulated expression of cloned genes in Candida albicans, a plasmid was constructed using the inducible promoter of the C . Albicans MAL2 gene . To demonstrate that the MAL2 promoter could regulate cloned genes placed under its control, a fusion construct was made with the coding sequence of the C . albicans URA3 gene . This plasmid was introduced into a Ura- strain of C . albicans using the process of restriction enzyme-mediated integration (REMI) . This procedure involves the transformation of the BamHI-linearized plasmid in the presence of BamHI enzyme . The majority of transformants generated contained insertions of the plasmid at chromosomal BamHI sites . All transformants examined were inducible for URA3 expression, which was determined by growth analysis and by measuring the level of URA3 gene product activity . The URA+ phenotype of the transformants was stable during growth under nonselective conditions . This system offers the advantages of stable transformation, easy recovery of integrated DNA, and inducible expression of genes in C . albicans.

Schweiz Rundsch Med Prax, 1996 Apr 16, 85(16), 520 - 5
{Candida albicans spondylitis: successful treatment with fluconazole . 2 case reports}; Rosler-Meier D et al.; Two patients suffering from candida albicans spondylitis in the lumbar spine are reported . Both had been operated on because of malignancy . One patient's clinical course was complicated by candida septicemia, and later on, serious candida endophthalmia developed . Conventional X-rays, combined with technetium scintigraphy, showed inflammatory destruction of the end plates of neighbouring vertebrae . Computerized tomography was done in the second patient, when an abscess was detected on both sides in the psoas muscle . Consecutively, operative removal and a spondylodesis were performed . Candida albicans could be isolated from removed discal material . Treatment by fluconazole for six months, in the first case in the beginning also by amphotericin B, lead to the healing of fungal spondylitis, accompanied by partial osseous consolidation.

FEMS Microbiol Lett, 1996 Apr 15, 138(1), 83 - 8
Analysis of the adaptive oxidative stress response of Candida albicans; Jamieson DJ et al.; Treatment of Candida albicans with low concentrations of either hydrogen peroxide or menadione (a superoxide generating agent) induces an adaptive response which protects cells from the lethal effects of a subsequent challenge with higher concentrations of these oxidants . Pre-treatment with either menadione or hydrogen peroxide is protective against cell killing by either oxidant . This suggests that the pathogenic yeast C . albicans (unlike the budding yeast Saccharomyces cerevisiae which has separate responses) possesses an adaptive response that responds to both these oxidants . In addition, we found that C . albicans showed a greater level of resistance to oxidants, both H2O2 and redox-cycling agents, compared to that observed with S . cerevisiae . In an attempt to characterise the oxidative stress response in more detail we have analysed the effect of oxidants on the activities of a number of enzymes with known antioxidant activity.

Biochemistry, 1996 Apr 9, 35(14), 4314 - 25
Delineation of an active fragment and poly(L-proline) II conformation for candidacidal activity of bactenecin 5; Raj PA et al.; Bactenecin 5 and its fragments {BN22 (1-22), BN16 (7-22), and BC24 (20-43)} were synthesized by solid-phase methods . Their antifungal activities on Candida albicans have been studied and compared with those of the native bactenecin 5 . The conformational preferences of these peptides in aqueous and nonaqueous solutions and in lipid vesicles were examined by circular dichroism . The highly active N-terminal fragment (BN16) was examined in aqueous solution using 500 MHz two-dimensional NMR . Bactenecin 5 and its fragments are potent candidacidal agents against C . albicans . The N-terminal fragments (BN22 and BN16) of bactenecin 5 are relatively more active than the C-terminal fragment BC24, especially at lower concentrations . The N-terminal region (7-22) which retains the activity of the whole molecule appears to be the functional domain for candidacidal activity . The CD spectra of bactenecin 5 and its fragments are reminiscent of the CD spectrum of poly(L-proline) type II structure in aqueous and nonaqueous solutions and also in lipid vesicles . The temperature dependence of NH chemical shifts and 1H/2H exchange effect on amide resonances suggest the absence of intramolecularly hydrogen-bonded NH groups . The coupling constant (JNH-CalphaH) values, conformational restriction offered by the Pro residues (phi = -60 degrees +/- 15 degrees), the set of medium- and short-range nuclear Overhauser effects observed for the active N-terminal fragment (BN16), and the restrained structure calculation using DIANA suggest that poly(L-proline) type II conformers of the peptide molecules could be significantly populated in aqueous solution . The ability of bactenecin peptides to induce disruption of lipid vesicles correlates well with their activity . Our results suggest that poly(L-proline) type II structure may, indeed, be the biologically active conformation for candidacidal activity of bactenecin peptides.

FEMS Microbiol Lett, 1996 Apr 1, 137(2-3), 269 - 73
Simultaneous carriage of Candida albicans strains from HIV-infected patients with oral candidiasis: multilocus enzyme electrophoresis analysis; Reynes J et al.; Genetic diversity of 160 Candida albicans isolates from the oral cavity of 16 HIV-infected adults prior to antifungal treatment was assessed using multilocus enzyme electrophoresis (10 C . albicans colonies were randomly chosen from each specimen culture) . 20 electrophoretic types were distinguished from the analysis of 21 enzyme loci (10 were polymorphic) . Five patients (31%) were found to be colonized by 2 or 3 genetically distinct strains . Nevertheless, in these five cases, one strain predominated (from 7 to 9 of the 10 colonies) . Some HIV + patients with oral candidiasis appear to be simultaneously infected with several genetically different C . albicans strains before antifungal treatment.

Pediatr Infect Dis J, 1996 Apr, 15(4), 340 - 4
In vitro lymphocyte blastogenic responses and cytokine production in sickle cell disease patients with acute pneumonia; Taylor SC et al.; BACKGROUND: Pulmonary infections continue to be a major cause of morbidity and mortality in patients with sickle cell disease (SCD) . METHODS: In this study cell-mediated immunity in vitro was evaluated in 62 SCD patients (62 steady state and 16 with acute pneumonia) and compared with 44 normal controls (30 healthy and 14 with acute pneumonia) . Lymphocyte blastogenic responses to phytohemagglutinin, tetanus toxoid and Candida albicans antigen were assessed in all subjects . In addition production of tumor necrosis factor, alpha- and gamma-interferon (IFN) were assayed . RESULTS: The results revealed comparable blastogenic responses to all three stimuli in all subjects except SCD patients with pneumonia . This group showed poor responses to all stimuli . The mean counts per minute were decreased 65 to 90% when compared with the other patients . Cytokine production of IFN-alpha and TNF was equivalent in all subjects . Conversely IFN-gamma production in both SCD groups, steady state (35 +/- 6 U/ml) and SCD with pneumonia (14 +/- 6 U/ml), was significantly decreased when compared with those in normal healthy controls (65 +/- 14 U/ml) and with pneumonia (48 +/- 17 U/ml) . On analysis of individual titers 15 of 62 (24%) steady state and 10 of 16 (63%) SCD patients with pneumonia were deficient in IFN-gamma production in vitro . CONCLUSIONS: Acute pulmonary infections seem to have a profound effect on cell-mediated immunity in SCD . IFN-gamma deficiency, along with quantitative and qualitative T cell abnormalities, may represent significant factors to explain the frequent and severe infections seen in SCD.

Antimicrob Agents Chemother, 1996 Apr, 40(4), 1044 - 7
Inhibition of 2,3-oxidosqualene-lanosterol cyclase in Candida albicans by pyridinium ion-based inhibitors; Goldman RC et al.; The N-(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecatrien-1- ylpyridinium and N-(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecatrien-1- ylpicolinium cations were evaluated for their ability to inhibit 2,3-oxidosqualene-lanosterol cyclase activity in Candida albicans . Both compounds inhibited fungal growth, were fungicidal, and resulted in the accumulation of squalene epoxide concurrent with a decrease in ergosterol, monomethyl sterols, and lanosterol, as was expected for the specific inhibition of 2,3-oxidosqualene-lanosterol cyclase activity . These compounds are electron-poor aromatic mimics of a monocyclized transition state or high-energy intermediate formed from oxidosqualene, which may explain their selective action.

Protein Sci, 1996 Apr, 5(4), 640 - 52
Structure of a secreted aspartic protease from C . albicans complexed with a potent inhibitor: implications for the design of antifungal agents; Abad-Zapatero C et al.; The three-dimensional structure of a secreted aspartic protease from Candida albicans complexed with a potent inhibitor reveals variations on the classical aspartic protease theme that dramatically alter the specificity of this class of enzymes . The structure presents: (1) an 8-residue insertion near the first disulfide (Cys 45-Cys 50, pepsin numbering) that results in a broad flap extending toward the active site; (2) a 7-residue deletion replacing helix hN2 (Ser 110-Tyr 114), which enlarges the S3 pocket; (3) a short polar connection between the two rigid body domains that alters their relative orientation and provides certain specificity; and (4) an ordered 11-residue addition at the carboxy terminus . The inhibitor binds in an extended conformation and presents a branched structure at the P3 position . The implications of these findings for the design of potent antifungal agents are discussed.

J Clin Microbiol, 1996 Apr, 34(4), 767 - 77
Most frequent scenario for recurrent Candida vaginitis is strain maintenance with "substrain shuffling": demonstration by sequential DNA fingerprinting with probes Ca3, C1, and CARE2; Lockhart SR et al.; The following three basic scenarios have emerged for the genetic relatedness of strains in recurrent vaginal candidiasis: strain maintenance without genetic variation, strain maintenance with minor genetic variation, and strain replacement . To test the frequency of each of the three scenarios, the genetic relatedness of Candida albicans isolates from each of 18 patients with recurrent infections was assessed by sequential DNA fingerprinting with the following three probes: the Ca3 probe; the C1 probe, a subfragment of the Ca3 probe which hybridizes to hypervariable genomic fragments; and the unrelated CARE2 probe . In each of the 18 patients with recurrent infections, the same strain was responsible for sequential infections, suggesting that the predominant scenario is strain maintenance . However, in 56% of these patients, the strain exhibited minor genetic variations in sequential infections . These changes were not found to be progressive . Rather, the changes suggest that substrains of an established infecting strain are shuffled in sequential infections . Results are also presented that in 45% of patients with recurrent infections, oral and vulvovaginal isolates were identical, in 35% they were highly related but not identical, and in 20% they were unrelated . These results differ markedly from those for commensal isolates simultaneously cultured from the oral cavity and vulvovaginal region of healthy individuals . Finally, it is demonstrated that in all eight cases in which C . albicans was isolated from both the male sexual partner of the patient with a recurrent infection and the patient, an isolate from the male partner was identical or highly related to the vulvovaginal strain . These results demonstrate that in patients with recurrent vulvovaginitis, a single strain usually dominates both in the different body locations of the patient and in the male partner and that it is maintained through sequential infections . However, in patients with recurrent infections, different substrains of the established clone dominate in an apparently random fashion, a process that we refer to as "substrain shuffling".

Indian Pediatr, 1996 Apr, 33(4), 299 - 303
Clinical profile and risk factors for oral candidosis in sick newborns; Gupta P et al.; OBJECTIVES: To provide the clinical profile and assess the significance of various risk factors contributing to the occurrence of oral candidosis in newborns . DESIGN: Case-control study . SETTING: Neonatal Intensive Care Unit (NICU) . SUBJECTS: Twenty newborns with oral candidosis and an equal number of age and weight matched controls . INTERVENTIONS: All cases of oral candidosis were treated with local application of 1% Clotrimazole . RESULTS: Oral candidosis was documented in 3.2% (20/650) cases in the NICU . Acute pseudomembranous candidosis was the most common presentation . The mean age of onset was 10.5 days . Candida albicans was isolated in 50% cases in addition to C . tropicalis, C . paratropicalis, C . krusei, C . glabrata and C . parapsilosis . On univariate analysis, male sex, birth asphyxia and prolonged antibiotic therapy had a significant correlation with occurence of oral candidosis in neonates . Out of these, birth asphyxia was the only factor significantly associated with oral candidosis (OR 8.09, 95% CI 1.34-48.8, p = 0.0226) on multivariate analysis . CONCLUSIONS: C . albicans was the predominant isolate in this series of oral candidosis . Clinical manifestations were evident in the second week of life and birth asphyxia was the most important associated perinatal eventPIP: During February-September 1992, all 650 infants admitted to the neonatal intensive care unit of the University College of Medical Sciences and G.T.B . Hospital were screened for oral thrush . A case control study was conducted to determine risk factors for oral candidiasis in newborns . The rate of oral candidiasis in this population was 3.2% (20 cases) . The most common pathogen was Candida albicans (50%) . All but 1 oral thrush case had acute pseudomembranous candidiasis . 75% of oral thrush cases were asymptomatic . Mean age of onset was 10.4 days (median, 9.5 days) . Clotrimazole solution was applied to oral lesions of all oral thrush cases . The multiple logistic regression revealed that birth asphyxia was the only significant factor responsible for oral thrush in newborns (odds ratio = 8.09; p = 0.0226) . These findings show that the most important perinatal event associated with oral thrush in newborns was birth asphyxia .

Yeast, 1996 Apr, 12(5), 501 - 4
Molecular cloning of a third chitinase gene (CHT1) from Candida albicans; McCreath KJ et al.; Here we report the complete nucleotide sequence of a third chitinase gene (CHT1) from the dimorphic human pathogen Candida albicans . The deduced amino acid (aa) sequence of Cht1 consists of 416 aa and displays 36% protein sequence similarity to chitinases Cht2 and Cht3, from C . albicans . Interestingly the domain structure of Cht1 is truncated when compared to the other chitinases of C . albicans and lacks a Ser/Thr-rich region.

Yeast, 1996 Apr, 12(5), 449 - 56
Candida albicans phosphatidylinositol synthase has common features with both Saccharomyces cerevisiae and mammalian phosphatidylinositol synthases; Antonsson BE et al.; Phosphatidylinositol (PI) synthase (cytidine 5'-diphospho (CDP)-1,2-diacyl-sn-glycerol:myo-inositol 3-phosphatidyltransferase, EC 2.7.8.11) was isolated from the microsomal cell fraction of Candida albicans . The Triton X-100 extracted enzyme was enriched 140-fold by affinity chromatography on CDP-diacylglycerol-Sepharose . The enzyme had a pH optimum at 9.5 in glycine/NaOH buffer . It had an absolute requirement for Mg2+ or Mn2+ and was inhibited by Ca2+ and Zn2+ . Maximal activity was at 0.2-0.6 mM-CDP-diacylglycerol, higher concentrations inhibited the enzyme . With 2'-deoxy-CDP-diacylglycerol as the lipid substrate, optimal activity was at 0.7 mM . The K(m) for myo-inositol was determined to be 0.55 mM . The optimal temperature for the PI synthase reaction was 55 degrees C . The C . albicans PI synthase shows differences to the Saccharomyces cerevisiae enzyme, such as activation by bivalent cations, inhibition by nucleotides, temperature optimum and activation energy, but also to the human PI synthase in preference for the lipid substrates, inhibition by nucleoside monophosphates and stabilization by Mn2+ and phospholipids.

FEMS Immunol Med Microbiol, 1996 Apr, 13(4), 311 - 6
A glucocorticoid antagonist, mifepristone affects anti-Candida activity of murine neutrophils in the presence of prednisolone in vitro and experimental candidiasis of prednisolone-treated mice in vivo; Abe S et al.; The effects of a glucocorticoid-antagonist, mifepristone on the suppressive action of prednisolone for anti-Candida activity of murine neutrophils were examined . Prednisolone suppressed inhibitory activity of neutrophils to mycelial growth of Candida albicans . This suppression was cancelled in the presence of 10(-7)-10(-6) M of mifepristone in vitro . Corresponding to this in vitro action, mifepristone protected prednisolone-treated mice from lethal C . albicans infection in vivo . These results suggest that glucocorticoid-induced vulnerability to Candida infection may be recovered or normalized by application of mifepristone.

Clin Exp Allergy, 1996 Apr, 26(4), 452 - 60
Enhanced IgE response to Candida albicans in postoperative invasive candidiasis; Savolainen J et al.; BACKGROUND: Invasive candidiasis is a life-threatening complication problem in post-operative and immunocompromized patients, e.g . those treated by intensive care . Candida is frequently cultured from the mucous membranes of hospital patients and fungal cultures offer little diagnostic help . Other diagnostic methods, such as blood cultures, serology and diagnostic imaging techniques produce results too late and, if positive, low sensitivity . OBJECTIVE: To study the value of Candida-specific antibodies, especially those of IgE class, in diagnosing invasive Candida infection . METHODS: The immunoglobulins IgE, IgG and IgM responses to antigens of Candida albicans in the sera of 14 patients with culture, biopsy and/or autopsy proven postoperative invasive candidiasis and of 11 colonized and 19 non-colonized operated patients were studied by mannan radioallergosorbent test (RAST), mannan enzyme-linked immunosorbent assay (ELISA) and immunoblotting . RESULTS: Detection of IgE antibodies to C . albicans polysaccharide (mannan) and protein antigens proved specific and sensitive in diagnostics of invasive candidiasis after major abdominal surgery . IgE rose early in the course of the infection and the method made a clear distinction between invasive infection and mucous colonization . Immunoblotting for protein antibodies was most sensitive while nitrocellulose-RAST for mannan antibodies was most specific . The combined use of immunoblotting and RAST increased the sensitivity and the specificity . Determinations of anti-Candida IgG and IgM antibodies had low sensitivity and specificity . CONCLUSION: Critically ill patients with invasive candidiasis develop IgE antibodies to Candida antigens probably because of disturbed TH1/TH2 responses . Determination of specific IgE antibodies can be used as a diagnostic aid in the early stage of invasive Candida infection.

Int J Oral Maxillofac Surg, 1996 Apr, 25(2), 136 - 44
Candida albicans: a review of its history, taxonomy, epidemiology, virulence attributes, and methods of strain differentiation; McCullough MJ et al.; The dimorphic yeast Candida albicans has been recognized as an increasingly important human pathogen particularly in immunocompromised hosts because of advanced age, infection or immunosuppressive therapy . This review outlines the history, taxonomy and epidemiology of this medically important yeast as well as discussing some of characteristics which are purported to be related to its virulence . Methods utilized for strain differentiation in the study of the epidemiologic relationship of members of this species are discussed.

Genitourin Med, 1996 Apr, 72(2), 98 - 102
Comparison of the efficacy and safety of oral fluconazole and topical clotrimazole in patients with candida balanitis; Stary A et al.; One hundred fifty seven men with candidal balanitis were entered in a randomised, open-label parallel-group multicentre study comparing efficacy and safety of a single oral 150-mg fluconazole-dose with clotrimazole applied topically twice daily for 7 days . Of 64 fluconazole and 68 clotrimazole treated patients who were evaluable at short term follow up, 92% and 91% respectively were clinically cured or improved . Candida albicans was eradicated in 78% and 83% of patients respectively . Median time to relief of erythema was 6 days for fluconazole and 7 days for clotrimazole . Twelve of 15 patients who had received previous topical therapy for balanitis said they preferred oral therapy . At the one month follow up visit, 24/36 and 29/33 patients in the two groups were clinically cured or improved . Nine in the fluconazole group experienced a relapse; 6 of these 9 patients reported previous episodes of this infection during the past year . Two patients in the clotrimazole group had a relapse; neither had a history of previous episodes . Mycological eradication was noted in 26/36 and 25/33 patients in the two groups . Both treatment regimens were well tolerated . Thus a single 150 mg dose of fluconazole was comparable in efficacy and safety to clotrimazole cream applied topically for 7 days when administered to patients with balanitis.

Chem Pharm Bull (Tokyo), 1996 Apr, 44(4), 785 - 92
Structure-activity relationships of 3-methyl and 3,3-dimethyl analogs of 2-(2,4-difluorophenyl)-3-(omega-substituted alkyl)sulfonyl-1-(1H-1,2,4-triazol-1-yl)-2-propanols; Miyauchi H et al.; 3-Methyl and 3,3-dimethyl analogs of 2-(2,4-difluorophenyl)-3-(omega-substituted alkyl)sulfonyl-1-(1H-1,2,4-triazol-1-yl)-2-propanols were synthesized and evaluated for their antifungal activities against Candida albicans and Aspergillus fumigatus . The 3,3-dimethyl analogs were found to have more potent activity both in vitro and in vivo than the corresponding 3-mono-methyl analogs . The prophylactic efficacy of the lead compounds against murine systemic candidiasis and aspergillosis was improved significantly by dimethylation of the 3-position.

Biomaterials, 1996 Apr, 17(7), 741 - 4
Effect of phagocytosis of pHEMA particles and of heat-killed Candida albicans on expression of carbohydrate-binding sites such as endogenous lectins in phagocytes; Smetana K Jr et al.; Phagocytosis of particles is an integral part of the defence system . Besides clearing the environment of foreign material this mechanism may affect the expression of physiologically relevant epitopes such as carbohydrate-binding sites, e.g . lectins . To determine the effect of particle design on the expression of such determinants in human monocytes and peritoneal macrophages either in suspension or after adherence, the binding of labelled (neo)glycoproteins was comparatively studied after exposure to poly (2-hydroxyethyl methacrylate) particles and heat-inactivated Candida albicans cells . A stimulatory effect of phagocytosis on extent of expression of binding sites for (neo)glycoproteins in phagocytic cells was observed . The levels of responsiveness varied according to the type of particle, adherence of the cell adding a further regulatory parameter . These results support the notion of a potential influence of the chemical structure and/or the form of an engulfed particle on phagocyte differentiation.

J Exp Med, 1996 Apr 1, 183(4), 1345 - 55
Impaired neutrophil response and CD4+ T helper cell 1 development in interleukin 6-deficient mice infected with Candida albicans; Romani L et al.; To define the role of interleukin (IL)6 in Candida albicans infection, IL-6 deficient mice were assessed for susceptibility to systemic or gastrointestinal infection, as well as for parameters of elicited T helper cell (Th) immunity . IL-6-deficient mice were more susceptible than wild-type mice to either type of infection caused by virulent C . albicans . In response to systemic challenge with a live vaccine strain of yeast, IL-6-deficient mice failed to mount Th1-associated protective immunity, but the resulting Th2-biased response could be redirected to the Th1 phenotype by IL-10 neutralization . Severe impairment of the macrophage and neutrophil response to infection was observed in IL-6-deficient mice, but administration of IL-6 would increase both neutrophil response and resistance to infection . IL-6 seems to oppose the Th2-promoting role of IL-10 in candidiasis, its early regulatory activity involving effects on neutrophil function.

J Prosthet Dent, 1996 Apr, 75(4), 426 - 31
Denture stomatitis: quantification of interleukin-2 production by mononuclear blood cells cultured with Candida albicans; Rodriguez-Archilla A et al.; Denture stomatitis is usually associated with the presence of yeast, particularly Candida albicans, and several bacteria . In this study mononuclear blood cells were grown in the presence of Candida albicans from a single colony, and interleukin-2 production induced in T lymphocytes was measured . Blood cells were from a population of patients with denture stomatitis and a control group of denture wearers without stomatitis . Induction of interleukin-2 production was correlated with factors that condition denture stomatitis, namely, isolation of Candida albicans in selective medium, age of the denture, and diabetes . Concentrations of interleukin-2 in supernatant and serum were also compared . Significant differences in interleukin-2 production were found between patients with denture stomatitis and controls . Statistical analysis demonstrated a significant association between isolation of Candida albicans and elevated interleukin-2 production in cultures from patients with and without denture stomatitis.

J Antibiot (Tokyo), 1996 Apr, 49(4), 380 - 5
Syntheses and antimicrobial activities of five-membered ring heterocycles coupled to indole moieties; Pereira ER et al.; Indole-substituted oxazolidinones, oxazolones, pyrrolidinone, imidazolidinone and imidazolones were synthesized . Their inhibitory potencies towards protein kinase C and protein kinase A were determined and their in vitro activities against Streptomyces chartreusis, Streptomyces griseus, Bacillus cereus, Candida albicans and Escherichia coli were examined . The inhibition of Streptomyces sporulation observed for some of them could not be linked to in vitro protein kinase C inhibition . All proved inactive against C . albicans but three of them exhibited a marked activity towards E . coli . This effect extends to other Gram-negative bacteria.

J Bacteriol, 1996 Apr, 178(8), 2416 - 9
Role of three chitin synthase genes in the growth of Candida albicans; Mio T et al.; The CHS2 and CHS3 genes of Candida albicans were disrupted . The double disruptant was still viable . Assessment of chitin and of calcofluor white resistance shows that CHS1 is responsible for septum formation and CHS3 is responsible for overall chitin synthesis otherwise . There were only small differences in virulence to immunocompromised mice of homozygous chs2 delta amd chs3 delta null mutants.

J Bacteriol, 1996 Apr, 178(8), 2320 - 7
Isolation and characterization of the GFA1 gene encoding the glutamine:fructose-6-phosphate amidotransferase of Candida albicans; Smith RJ et al.; Glutamine:fructose-6-phosphate amidotransferase (glucosamine-6-phosphate synthase) catalyzes the first step of the hexosamine pathway required for the biosynthesis of cell wall precursors . The Candida albicans GFA1 gene was cloned by complementing a gfa1 mutation of Saccharomyces cerevisiae (previously known as gcn1-1; W . L . Whelan and C . E . Ballou, J . Bacteriol . 124:1545-1557, 1975) . GFA1 encodes a predicted protein of 713 amino acids and is homologous to the corresponding gene from S . cerevisiae (72% identity at the nucleotide sequence level) as well as to the genes encoding glucosamine-6-phosphate synthases in bacteria and vertebrates . In cell extracts, the C . albicans enzyme was 4-fold more sensitive than the S . cerevisiae enzyme to UDP-N-acetylglucosamine (an inhibitor of the mammalian enzyme) and 2.5-fold more sensitive to N3-(4-methoxyfumaroyl)-L-2,3-diaminopropanoic acid (a glutamine analog and specific inhibitor of glucosamine-6-phosphate synthase) . Cell extracts from the S . cerevisiae gfa1 strain transformed with the C . albicans GFA1 gene exhibited sensitivities to glucosamine-6-phosphate synthase inhibitors that were similar to those shown by the C . albicans enzyme . Southern hybridization indicated that a single GFA1 locus exists in the C . albicans genome . Quantitative Northern (RNA) analysis showed that the expression of GFA1 in C . albicans is regulated during growth: maximum mRNA levels were detected during early log phase . GFA1 mRNA levels increased following induction of the yeast-to-hyphal-form transition, but this was a response to fresh medium rather than to the morphological change.

Int Arch Allergy Immunol, 1996 Apr, 109(4), 334 - 43
Candida albicans mannan-specific, delayed hypersensitivity down-regulatory CD8+ cells are genetically restricted effectors and their production requires CD4 and I-A expression; Li SP et al.; There is considerable controversy over the induction and activity of down-regulatory cells active in various antigen-specific and antigen-nonspecific systems . We have been studying the nature of such cells in a Candida albicans mannan (MAN)-specific system for some time and report here the requirements for CD4+ and I-A+ cells during the inductive phase for the development of CD8+ effector cells, as well as the requirement for genetic compatibility for effector activity of CD8+ cells . Since we have shown previously that CD8+ down-regulatory cells were present in spleens of MAN-treated mice 4 days following the administration of MAN to naive mice, as determined by their ability to suppress delayed hypersensitivity (DH) when transferred to immunized recipients, we treated mice with monoclonal antibodies specific for CD4 and I-A at various times before, with or after the administration of MAN to assess the role of CD4+ and I-A+ cells in the development of the CD8+ effector cell . Both anti-CD4 and anti-I-A given before or up to 30 h after the administration of MAN abrogated the ability of splenocytes from MAN-treated mice to down-regulate MAN-specific DH in immunized recipients . Moreover, transfers of down-regulatory cells between H-2-incompatible strains of mice, specifically CBA/J and BALB/cByJ, provided evidence that the effector cell for the down-regulatory activity was also restricted genetically in its activity . Taken together, the data presented indicate that genetically compatible cells are required for both the inductive and effector stages of down-regulation of MAN-specific DH, suggesting that cell-cell cooperation is required for both stages and that CD4+ cells are required in a pathway leading to the development of the CD8+ effector cell.

Curr Genet, 1996 Apr, 29(5), 441 - 5
Aromatic amino-acid biosynthesis in Candida albicans: identification of the ARO4 gene encoding a second DAHP synthase; Pereira SA et al.; The primary step in the aromatic amino-acid biosynthetic pathway in Saccharomyces cerevisiae is catalyzed by two redundant isozymes of 3-deoxy-d-arabinoheptulosonate-7-phosphate (DAHP) synthase, either of which alone is sufficient to permit growth on synthetic complete media lacking aromatic acids (SC-Aro) . The activity of one isozyme (encoded by the ARO3 gene) is feedback-inhibited by phenylalanine, whereas the activity of the other isozyme (encoded by the ARO4 gene) is feedback-inhibited by tyrosine . Transcription of both genes is controlled by GCN4 . We previously cloned the ARO3 gene from the opportunistic pathogen Candida albicans and found that: (1) it can complement an aro3 aro4 double mutation in S . cerevisiae, an effect inhibited by excess phenylalanine; and (2) its expression is induced in response to amino-acid deprivation, consistent with the presence of two putative GCN4-responsive promoter elements (Pereira and Livi 1993, 1995) . To determine whether other DAHP synthases exist in C . albicans, we have constructed a homozygous aro3-deletion mutant strain . Such a mutant was found to be phenotypically Aro+, i . e., capable of normal growth on SC-Aro media, suggesting the presence of at least one additional isozyme . To confirm this result, a 222-bp DNA fragment was amplified by the polymerase chain reaction (PCR) from genomic DNA prepared from the homozygous aro3-deletion mutant, using a degenerate primer based on a conserved N-terminal region of Aro3p plus a degenerate comeback primer encoding a conserved region of the protein that lies within the deleted portion of the gene . The nucleotide sequence of this PCR fragment predicts a 74-amino acid DAHP synthase-related protein which shows strong homology to Aro3p from S . cerevisiae and C . albicans, but even greater homology (78% identity) to S . cerevisiae Aro4p . We conclude that cells of C . albicans contain a second Aro4p-related DAHP synthase.

J Med Microbiol, 1996 Apr, 44(4), 311 - 6
In-vitro proteinase production by oral Candida albicans isolates from individuals with and without HIV infection and its attenuation by antimycotic agents; Wu T et al.; In-vitro proteinase production by oral Candida albicans isolates from patients with and without HIV infection (18 isolates from each group) was assessed by image analysis of a plate assay, with bovine serum albumin (BSA) as a substrate . The effect of sub-minimal inhibitory concentrations (sub-MICs) of nystatin, amphotericin B, clotrimazole and miconazole on in-vitro proteinase production by these yeast isolates was also investigated . Proteinase production by C . albicans isolates from patients with HIV infection was significantly greater than production by those from individuals without infection . All 18 isolates from HIV-infected individuals produced proteinase, in comparison to 56% of isolates from uninfected individuals . Pre-exposure of C . albicans isolates (seven proteinase producers from each group) to 1/4 and 1/16 MICs of nystatin, amphotericin B, clotrimazole and miconazole resulted in decreased proteinase production in all isolates tested . However, after exposure to the four antimycotic agents, proteinase production was decreased to a significantly greater extent in isolates from uninfected individuals than in those with HIV disease . Furthermore, when the relative concentration effect of antimycotic agents on proteinase production was compared, C . albicans isolates from the HIV-free group demonstrated a salient dose-response relationship compared with the HIV-infected group . These results indicate that C . albicans from patients with HIV infection are significantly more proteolytic than those from individuals without the infection, and that polyenes and imidazoles curtail the proteolytic activity of all C . albicans isolates, albeit to a lesser extent in those from HIV-infected patients . It appears that HIV disease favours oral colonisation by more proteolytic C . albicans isolates, with resilient proteolytic activity.

J Med Microbiol, 1996 Apr, 44(4), 277 - 83
Immune responsiveness in a rat model for type II diabetes (Zucker rat, fa/fa): susceptibility to Candida albicans infection and leucocyte function; Plotkin BJ et al.; There is a causal relationship between obesity-associated diabetes and an increased risk of infection . The ability of obese (fa/fa) Zucker rats, a model for non-insulin-dependent diabetes mellitus (NIDDM), to clear Candida albicans from the circulation and tissues was compared to that of lean (Fa/fa, Fa/Fa) Zucker rat controls as a measure of immune function . The ID50 necessary to establish tissue colonisation in lean Zucker rats was 1.18 log10 times greater than that determined for the obese Zucker rats . Nine days after intravenous (i.v.) injection of a yeast suspension, the organs of obese rats had a 10-fold greater yeast/g organ burden than did lean rats . The kidney was determined to be the primary target organ for colonisation . Germ-tube formation by C . albicans occurred at a rate 1.5 times faster in serum from obese rats than in serum from lean rats . Peritoneal polymorphonuclear leucocytes, resident macrophages and thioglycollate-elicited macrophages from lean Zucker rats displayed a significantly higher ability to kill ingested yeast cells than analogous cell populations from obese Zucker rats.

Infect Immun, 1996 Apr, 64(4), 1379 - 84
Resistance to platelet microbicidal protein results in increased severity of experimental Candida albicans endocarditis; Yeaman MR et al.; Thrombin-induced platelet microbicidal protein (tPMP) exerts potent in vitro microbicidal activity against pathogens commonly found in the bloodstream, including Staphylococcus aureus, Staphylococcus epidermidis, and Candida albicans . Localized platelet release of tPMP may be important in defense against infections involving the vascular endothelium caused by tPMP-susceptible organisms . In contrast, pathogens capable of surviving in the presence of tPMP could then exploit the platelet as an adhesive surface for attachment to damaged endothelium . To examine these hypotheses, we derived a tPMP-resistant (tPMP(r)) C . albicans strain from its tPMP-sensitive (tPMP(s)) parental strains were equivalent in vitro as assessed by genotyping (electrophoretic karyotype and restriction endonuclease analysis of genomic DNA), biotyping, germination, platelet aggregation, adherence to vascular endothelial cells, and growth characteristics . In addition, the tPMP(r) phenotype was stable following multiple in vitro and in vivo passages . We then investigated the in vivo relevance of tPMP susceptibility on endovascular infection using a rabbit model of endocarditis and hematogenous dissemination . Rabbits with transaortic catheters (n = 15 in each group) were challenged with either the tPMP(s) or tPMP(r) C . albicans strain . All rabbits developed C . albicans-induced endocarditis, as determined by the presence of infected vegetations . In rabbits challenged with tPMP(s) strain (P < 0.001) . These results were seen in the absence of differences in either initial adherence of strains to cardiac valves or vegetation weights . Furthermore, although these C . albicans strains induced equivalent rates and extent of hematogenous renal infection, only the tPMP(r) strain disseminated hematogenously to the spleen (15 of 15 rabbits) versus 0 of 15 {tpmp(s) strain}; P < 0.0001) . Thus, tPMP(r) C . albicans caused more-severe endocarditis and produced greater metastatic sequelae than the tPMP(s) counterpart.

Yeast, 1996 Mar 30, 12(4), 361 - 8
Candida albicans homologue of GGP1/GAS1 gene is functional in Saccharomyces cerevisiae and contains the determinants for glycosylphosphatidylinositol attachment; Vai M et al.; The GGP1/GAS1/CWH52 gene of Saccharomyces cerevisiae encodes a major exocellular 115 kDa glycoprotein (gp115) anchored to the plasma membrane through a glycosylphosphatidylinositol (GPI) . The function of gp115 is still unknown but the analysis of null mutants suggests a possible role in the control of morphogenesis . PHR1 gene isolated from Candida alibicans is homologous to the GGP1 gene . In this report we have analysed the ability of PHR1 to complement a ggp1 delta mutation in S . cerevisiae . The expression of PHR1 controlled by its natural promoter or by the GGP1 promoter has been studied . In both cases we have observed a complete complementation of the mutant phenotype . Moreover, immunological analysis has revealed that PHR1 in budding yeast gives rise to a 75-80 kDa protein anchored to the membrane through a GPI, indicating that the signal for GPI attachment present in the C . albicans gene product is functional in S . cerevisiae.

J Med Chem, 1996 Mar 15, 39(6), 1227 - 35
Molecular modeling of azole antifungal agents active against Candida albicans . 1 . A comparative molecular field analysis study; Tafi A et al.; A series of 56 azole antifungal agents belonging to chemically diverse families related to bifonazole, one of the antimycotic drugs of clinical use, were investigated using the comparative molecular field analysis (CoMFA) paradigm . The studied compounds, which have been already synthesized and reported to be active in vitro against Candida albicans, were divided into a training set and a test set . The training set consisted of 40 molecules from all the different structural classes . Due to the lack of experimental structural data on these derivatives, molecular mechanics techniques were used to obtain putative active conformations for all the compounds . the correctness of this molecular modeling work was confirmed a posteriori by comparison with structural data of the analog 2w obtained by X-ray crystallographic analysis (Massa, S.; et al . Eur . J . Med . Chem . 1992, 27, 495-502) . Two different alignment rules of the training set molecules were used in this study and are based on the assumption that according to published results on azole antifungal agents, all the studied compounds exert their inhibitory activity through the coordination of their azole moiety to the protoporphyrin iron atom of the fungal lanosterol 14alpha-demethylase enzyme . The predictive ability of each resultant CoMFA model was evaluated using a test set consisting of 16 representative compounds that belong to all the different structural classes . The best 3D-quantitative structure-activity relationship model found yields significant cross-validated, conventional, and predictive r2 values equal to 0.57, 0.95, and 0.69, respectively . The average absolute error of predictions of this model is 0.30 log units, and the structural moieties of the studied antifungal agents which are thought to contribute to the biological activity were identified . The predictive capability of this model could be exploited in further synthetic studies on antifungal azoles . Furthermore, the results obtained by using two different alignments of the inhibitors suggest that the binding mode of these molecules involves both a coordination to the iron protoporphyrin atom and an additional, likewise relevant, hydrophobic interaction with the active site.

Zhonghua Zheng Xing Shao Shang Wai Ke Za Zhi, 1996 Mar, 12(2), 132 - 4
{Mycethemia: an analysis of 56 cases}; Zhang H et al.; Mycethemia was diagnosed in 56 burn patients between 1958 and 1992 . The overall incidence was 0.39% and the mortality rate was 39.29% . Candida albicans, constituting for 75% of cases, was most common encountered . The incidence was 0.26% and the mortality 72.73% in years before 1985, while that in the years after 1985 were 0.58% and 17.65%, respectively . The differences are of significance (P < 0.01) predisposing . The clinical characteristics were summed up, and the causes, clinical diagnosis and treatment were studied and discussed.

Chin Med Sci J, 1996 Mar, 11(1), 45 - 8
Effects of systemic fluconazole therapy on in vitro adhesion of Candida albicans to buccal epithelial cells and changes of the cell surface proteins of the epithelial cells; Wu S et al.; This paper presented the effects of systemic fluconazole therapy via intravenous (IV) and oral (PO) administrations on the adhesion of Candida albicans (C . albicans) to the buccal epithelial cells (BEC) from five treated patients with three candidosis, one mucormycosis and one sporotrichosis and at the same time, an analysis of the cell surface proteins involving candidal adherent receptor in the BEC of the patients in the course of 7 days were exposed to 3H-leucine radiolabeled C . albicans for in vitro candidal adherent assay, and the BEC from first intake day and the last intake day of the patients were extracted by dithiothreitol (DTT)-iodoacetamide treatment for SDS-PAGE . These results indicate that the systemic fluconazole therapy results in the inhibitory effect of candidal adhesion to BEC of treated patients to prevent them from oral candidosis for a prolonged time, which is based on the absent surface protein (35 KDa) of the BEC.

Microbiology, 1996 Mar, 142 ( Pt 3), 485 - 92
Candida albicans has a cell-associated ferric-reductase activity which is regulated in response to levels of iron and copper; Morrissey JA et al.; For survival, pathogenic organisms such as Candida albicans must possess an efficient mechanism for acquiring iron in the iron-restricted environment of the human body . C . albicans can use iron from a variety of sources found within the host . However, it is not clear how biologically active ferrous iron is obtained from these sources . One strategy adopted by some organisms is to reduce iron extracellularly and then specifically transport the ferrous iron into the cell . We have shown that clinical isolates of C . albicans do have a cell-associated ferric-reductase activity . The determination of ferric-reductase activity of cells growing exponentially in either low- or high-iron media over a period of time indicated that C . albicans reductase activity is induced when in low-iron conditions . Moreover, we have demonstrated that C . albicans reductase activity is also regulated in response to the growth phase of the culture, with induction occurring upon exit from stationary phase and maximal levels being reached in early exponential stage irrespective of the iron content of the medium . These results suggest that C . albicans reductase activity is regulated in a very similar manner to the Saccharomyces cerevisiae ferric-reductase . Iron reduction and uptake in S . cerevisiae are closely connected to copper reduction, and possibly copper uptake . In this report we show that iron and copper reduction also appear to be linked in C . albicans . The ferric-reductase activity is negatively regulated by copper . Moreover, quantitative cupric-reductase assays indicated that C . albicans is capable of reducing copper and that this cupric-reductase activity is negatively regulated by both iron and copper . This is the first report that C . albicans has an iron- and copper-mediated ferri-reductase activity.

Clin Infect Dis, 1996 Mar, 22(3), 462 - 6
Failure of systemic empirical treatment with amphotericin B to prevent candidemia in neutropenic patients with cancer; Blumberg EA et al.; We undertook a retrospective review of all patients with hematologic malignancies in whom candidemia developed during chemotherapy-induced neutropenia in 1989 and 1990 . Candidemia developed in 11 patients; five were receiving therapeutic doses of amphotericin B at the time of infection . Disseminated infection occurred in 2 of 5 patients with breakthrough infection and 3 of 6 patients with candidemia before receipt of amphotericin B . Among patients with breakthrough candidemia there was a trend toward more-prolonged neutropenia prior to infection (P = .069), but otherwise they were indistinguishable from other candidemic patients with regard to risk factors for candidemia . Amphotericin B-susceptible Candida albicans was isolated from two patients and Candida krusei from three patients with breakthrough infection . All patients were treated with amphotericin B; all breakthrough infections responded to treatment . Neutropenic patients with breakthrough candidemia were clinically similar to those whose candidemia preceded amphotericin B therapy, and there was no increase in morbidity and mortality among individuals with breakthrough infection.

Antimicrob Agents Chemother, 1996 Mar, 40(3), 816 - 8
Bleomycin therapy of experimental disseminated candidiasis in mice; Graybill JR et al.; Bleomycin, an antineoplastic agent, was found to be very effective in vitro against a variety of fungi, including Candida albicans . Mice were infected with C . albicans intravenously and then treated with various doses of bleomycin . No efficacy was shown by either prolongation of survival or reduction of tissue counts.

Antimicrob Agents Chemother, 1996 Mar, 40(3), 588 - 94
Susceptibilities of Candida spp . to antifungal agents visualized by two-dimensional scatterplots of relative growth; Odds FC et al.; The growth of 811 clinical yeast isolates in the presence of single concentrations of antifungal agents was measured spectrophotometrically and expressed as a percentage of growth in inhibitor-free control cultures . Two-dimensional scatterplots of the relative growth data allowed for the simple visual determination of some susceptibility trends, including correlations in relative growth between different agents and in relative susceptibilities between different yeast species . A positive susceptibility correlation was found for relative growth results with the azole antifungal agents fluconazole, itraconazole, and ketoconazole for 504 Candida albicans isolates . The relative growth scatterplots for fluconazole versus itraconazole showed that 50 (9.9%) of 504 C . albicans isolates were outliers with respect to the 95% confidence limits for a line of correlated relative growth established with an initial test panel of 59 isolates of this species . The outlying isolates were relatively less susceptible to fluconazole than to itraconazole under the conditions of the test . Most of the outliers were received in 1993 and 1994; only 3.9% of the isolates received in 1991 and 1992 and 1.7% of the isolates received before 1991 showed this differential susceptibility . In addition, most of the outliers came from patients with human immunodeficiency virus infections . The relative growth scatterplots confirmed the known high susceptibility of most Candida parapsilosis isolates to both fluconazole and itraconazole and the specifically low susceptibility of Candida krusei isolates to fluconazole . The scatterplots also illustrated a tendency towards lower (and correlative) relative growth among oral isolates obtained from AIDS patients who responded to azole antifungal treatment than among isolates from clinical nonresponders.

J Mol Med, 1996 Mar, 74(3), 135 - 42
The role of Candida albicans secreted aspartic proteinase in the development of candidoses; Hoegl L et al.; Although Candida albicans infections in humans are increasingly frequent, our understanding of the host-parasite relationship is limited . The secreted aspartic proteinase of C . albicans was first described in 1965 and has proved to be a major factor in virulence . This enzyme belongs to the class of aspartic proteinases which includes pepsin and renin in humans . Although found in some fungi, secreted aspartic proteinase is rare in these organisms . While the existence of several isoenzymes may not be fully established, it is now obvious that at least seven different genes encode for secreted aspartic proteinase . Within Candida cells it is located in membrane-bound vesicles . Upon fusion of these subcellular structures within the plasma membrane, the enzyme is released to the environment . In the context of human mucosal diseases it is responsible both for adhesion and invasion . Strains from HIV-infected patients with oral candidosis generally exhibit higher enzymatic activity than control strains . In future secreted aspartic proteinase may prove a prime target for new types of antimycotics.

Epidemiology, 1996 Mar, 7(2), 182 - 7
Risk factors for vulvovaginal candidiasis: a case-control study among university students; Geiger AM et al.; Vulvovaginal candidiasis (VVC) is a common inflammatory condition caused by vaginal overgrowth of Candida albicans . Typical symptoms include pruritus and discharge . To test the association between several hypothesized risk factors and VVC, we conducted a case-control study among university students, with both clinic and population controls . Symptomatic, culture-proven VVC was associated with receptive oral sex twice or more in the previous 2 weeks {vs not at all, odds ratio (OR) = 3.5; 95% confidence interval (CI) = 1.7-7.0}; oral contraceptive use (OR = 1.8; 95% CI = 0.95-3.6); spermicide use (OR = 3.3; 95% CI = 1.6-6.8); a prior diagnosis of VVC in the previous year (OR = 3.0; 95% CI = 1.5-5.9); and black (OR = 6.8; 95% CI = 3.1-15) and "other" race (OR = 2.2; 95% CI = 1.0-4.6) . Estimates are from a cases vs population controls logistic regression model including all five variables; results for cases vs clinic controls were similar . After adjusting for these factors, many other hypothesized risk factors, such as antibiotic use, menstrual hygiene practices, and vaginal intercourse, had little association with VVC.

Mol Microbiol, 1996 Mar, 19(5), 1107 - 16
Use of synthetic peptides to confirm that the Pseudomonas aeruginosa PAK pilus adhesin and the Candida albicans fimbrial adhesin possess a homologous receptor-binding domain; Yu L et al.; Pseudomonas aeruginosa PAK pili and Candida albicans fimbriae are adhesins present on the microbial cell surfaces which mediate binding to epithelial cell-surface receptors . The receptor-binding domain (adhesintope) of the PAK pilus adhesin has been shown previously to reside in the carboxy-terminal disulphide-bonded region of P . aeruginosa pilin (PAK128-144) . The delineation of the C . albicans fimbrial adhesintope was investigated in these studies using synthetic peptides which correspond to the whole (PAK128-144) or part of (PAK134-140) adhesintope of the PAK pilus and their respective anti-peptide antisera and biotinylated PAK pili (Bt-PAK pili), fimbriae (Bt-fimbriae), P . aeruginosa whole cells (Bt-P . aeruginosa) and C . albicans whole cells (Bt-C . albicans) . The results from these studies confirmed that a structurally conserved motif akin to the PAK(128-144) peptide sequence is present in C . albicans fimbrial adhesin and that the seven-amino-acid residue PAK(134-140) sequence plays an important role in forming the adhesintope for both P . aeruginosa PAK pilus and C . albicans fimbrial adhesins.

Dev Comp Immunol, 1996 Mar-Apr, 20(2), 97 - 104
Evidence for circulating hemocyte proliferation in the shrimp Penaeus japonicus; Sequeira T et al.; We have observed by flow cytometric analysis that a small but consistent percentage of circulating hemocytes in phases S, G2 and M of the cell cycle can be detected in Penaeus japonicus . Significantly increased percentages of proliferating hemocytes (approximately three-fold increase) were observed after stimulation by lipopolysaccharide (LPS), p43 (an immunosuppressive lymphocyte mitogenic protein produced by Candida albicans) and a combination of LPS and p43 . Moreover, an approximately six-fold increase in the percentage of proliferating hemocytes was also observed in animals infected with Fusarlum opp., as compared to non-infected shrimps . Furthermore, {3H} thymidine uptake in circulating hemocytes was 26 times greater in LPS stimulated than in non-stimulated shrimps . The present study suggests that circulating hemocytes of the shrimp P . japonicus can divide in vivo and that proliferation can be increased significantly after mitogenic or infectious stimulation.

Mycoses, 1996 Mar-Apr, 39(3-4), 111 - 4
Quantitative screening for fluconazole-amphotericin B antagonism in several Candida albicans strains by a comparative agar diffusion assay; Scheven M et al.; Antagonism between fluconazole (FCZ) and amphotericin B (AMB) was determined with an agar diffusion technique using series of agar plates containing no or 10 mgl-1 FCZ (comparative diffusion assay) . Serial dilutions of AMB produced concentration-dependent inhibition zones that varied between the two agar plate series . This technique served as screening method to determine FCZ-AMB interactions in 18 Candida albicans strains . The critical concentrations of AMB were enhanced 1.33- to 7.0-fold by FCZ . The critical time, T0, was reduced by half by FCZ.

Mycoses, 1996 Mar-Apr, 39(3-4), 103 - 10
Fungicidal activity of latex sap from Carica papaya and antifungal effect of D(+)-glucosamine on Candida albicans growth; Giordani R et al.; Carica papaya latex sap inhibits the growth of Candida albicans when added to a culture during the exponential growth phase . Approximately 60% was achieved . This fungistatic effect is the result of cell wall degradation due to a lack of polysaccharidic constituents in the outermost layers of the fungal cell wall and release of cell debris into the culture medium . When C . albicans was cultured on medium supplemented with D(+)-glucosamine, an inhibitor of N-acetyl-beta-D-glucosaminidase, growth was inhibited (34%) in a similar manner . Addition of D(+)-glucosamine during the exponential growth phase also had a fungistatic effect (26%) . The modes of action of C . papaya latex and of D(+)-glucosamine in cell wall breakdown are discussed.

J Med Vet Mycol, 1996 Mar-Apr, 34(2), 149 - 52
Comparisons of the susceptibilities of planktonic and adherent Candida albicans to antifungal agents: a modified XTT tetrazolium assay using synchronised C . albicans cells; Hawser S; Adhesion of synchronised yeast-phase Candida albicans cells to tissue culture plastic, and the susceptibility of planktonic and adherent cells to antifungal agents, was investigated using a modified tetrazolium (XTT) assay . MIC data demonstrated that ketoconazole and amphotericin B were highly active against planktonic C . albicans yeast-phase cells . XTT tetrazolium assays permitted comparisons of MIC values with XTT formazan IC50 and IC80 (percentage inhibitory concentrations); IC50 and IC80 values for amphotericin B and ketoconazole were similar . Furthermore, IC50 and IC80 values for 24 h incubation with antifungal agent were typically higher than corresponding IC50 and IC80 values for 48 h incubation . Furthermore, in comparison to values for planktonic Candida cells, adherent cells were typically less susceptible to amphotericin B and ketoconazole . For example, with increasing incubation time following the initial adhesion period, cells became progressively less susceptible to amphotericin B and ketoconazole: 24 h (P < 0.05) and 48 h (P < 0.001) . Furthermore, other azoles showed the same activities compared with ketoconazole against both planktonic and adherent cells . Overall, the data demonstrate the usefulness of the XTT tetrazolium assay in describing comparisons of the susceptibility profiles for both planktonic and adherent synchronous yeast phase C . albicans in vitro.

J Med Vet Mycol, 1996 Mar-Apr, 34(2), 99 - 104
Systemic candidiasis in Sprague-Dawley rats; Schmidt A; A reproducible model of a generalized Candida albicans infection was established in rats to allow a precise evaluation of the efficacy of antifungal compounds . In contrast to the intravenous C . albicans model in mice, which serves as a primary model for in vivo efficacy studies of antimycotic compounds, the infectious process in Sprague-Dawley rats is more severely spread into organs other than the kidneys, such as brain, heart, liver, lung, retina and spleen . Apart from a severe granulomatous nephritis beginning 1 day after infection, we observed a severe pneumonitis 3 days after infection with a mass of extravasal erythrocytes in the interstitium and the alveolar space . In addition, multiple nodular lesions could be observed in the brain, heart, liver, retina and spleen on the first day after infection . Lethality was 100% within 1 week, the majority of deaths occurring from 5 to 7 days . Antifungal therapy with amphotericin B or fluconazole led to long-term survival over 4 months, which could not be achieved in mice.

J Med Vet Mycol, 1996 Mar-Apr, 34(2), 91 - 7
Dynamic expression of cell wall proteins of Candida albicans revealed by probes from cDNA clones; Alloush HM et al.; Five cDNA clones were selected from the positive clones detected by screening a germ tube expression library constructed in lambda gt11 with rabbit antisera raised against cell wall extracts of Candida albicans . The selected clones were amplified and used to obtain affinity purified antibodies by eluting from the expressed proteins that had been previously transferred onto nitrocellulose discs . The antibodies obtained were used as probes in immunoblots of the cell wall extracts separated by denaturing polyacrylamide electrophoresis . A single protein band was detected for each clone . Detection of products of the cloned sequences varied according to the extraction procedure and/or cell morphology . These products included bands exhibiting apparent molecular weights of 40, 58, 68 and 70 kDa present in beta-mercaptoethanol (beta ME) extracts from both yeast and germ tubes, and a 30 kDa beta ME extracted protein specific for germ tubes . The expression of these products at the cell surface was confirmed by indirect immunofluorescence . Expression of the mRNAs of the different cDNA clones varied according to growth- and morphology-related factors and showed no direct correlation between expression and presence in the cell wall . These observations suggest that complex mechanisms are involved in the regulation and expression of cell surface components of C . albicans.

Diagn Microbiol Infect Dis, 1996 Mar, 24(3), 161 - 4
Candida albicans osteomyelitis of the zygomatic bone . A distinctive case with a possible peculiar mechanism of infection and therapeutic failure with fluconazole; Arranz-Caso JA et al.; This report describes a distinctive case of zygomatic candidiasic osteomyelitis in a diabetic patient with oral candidiasis and malar ulceration secondary to topic 5-fluoroacil toxicity that eventually exposed part of the underlying bone . The mechanism of infection may have been self-inoculation of spores from muguet plaques on the oral mucosa to the exposed bone tissue by hand contact . Such a mechanism of bone infection probably should be considered in patients who frequently have oral candidiasis (diabetes, malignancies, and HIV infection) and open lesions of the skin and soft tissues . Treatment with fluconazole was ineffective, but amphotericin B was curative.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1996 Mar, 81(3), 291 - 6
Prophylaxis of candidiasis in patients with leukemia and bone marrow transplants; Epstein JB et al.; OBJECTIVES . The increased risk for systemic fungal infection and the potential fatal consequences of disseminated candidiasis in bone marrow transplant patients has prompted study of prophylaxis and early treatment of candida colonization and infection . STUDY DESIGN . Patients with leukemia who received fluconazole prophylaxis were compared with a concurrent group of patients not given prophylaxis for fungal organisms . RESULTS . A trend to reduction of oropharyngeal colonization by Candida albicans was seen (p = 0.07) although no significant differences in systemic candidiasis were seen . In patients with documented systemic candidiasis, oral colonization was present and systemic infection was identified after the development of ulcerative oral mucositis . CONCLUSIONS . Our results support the potential of fluconazole to reduce oropharyngeal colonization by Candida albicans, however, we did not show prophylaxis of oral candidiasis or systemic candidiasis . These findings and reports of fluconazole-resistant candidal species and a rising number of cases of infection as a result of Candida krusei indicate the need for further studies of prophylaxis of candidal infection in patients who are anticipated to develop profound neutropenia.

J Invest Dermatol, 1996 Mar, 106(3), 549 - 52
Cholesterol sulfate protects Candida albicans from inhibition by sphingosine in vitro; Payne CD et al.; Sphingosine is known to have potent biological activity, including pronounced anti-microbial action in vitro against Candida albicans and some bacteria . Several sphingosine bases are present in stratum corneum at concentrations several orders of magnitude above those in other tissues . Sphingosine forms an undissociated salt with organic sulfates, however, so that the free sphingosine in the epidermis may be inactivated by the cholesterol sulfate known to be present . To investigate this hypothesis, C . albicans was grown in cultures with graded concentrations of sphingosine added in ethanol . In 1% ethanol, 0.1-100 microgram/ml sphingosine completely prevented growth of the organism for 12 h . All cultures eventually entered log-phase growth and reached limiting density at a rate inversely proportional to sphingosine concentration . When sphingosine was added, together with an equimolar amount of cholesterol sulfate, there was no delay in the onset of growth of the yeast and the rate of growth and final density were similar to control cultures . These results demonstrate that natural ratios of cholesterol sulfate neutralize the anti-microbial activity of sphingosine in vitro . In the epidermis, endogenous cholesterol sulfate is hydrolyzed by sterol sulfatase at the skin surface, where the released sphingosine may resist microbial colonization of the stratum corneum . This mechanism for liberating anti-microbial sphingosine base only at the skin surface may protect the viable epidermis against known cytotoxic effects of free sphingosine.

Infect Immun, 1996 Mar, 64(3), 891 - 6
Blood group glycolipids as epithelial cell receptors for Candida albicans; Cameron BJ et al.; The role of glycosphingolipids as possible epithelial cell receptors for Candida albicans was examined by investigating the binding of biotinylated yeasts to lipids extracted from human buccal epithelial cells and separated on thin-layer chromatograms . Binding was visualized by the addition of 125I-streptavidin followed by autoradiography . Five C . albicans strains thought from earlier work to have a requirement for fucose-containing receptors all bound to the same three components in the lipid extract . A parallel chromatogram overlaid with biotinylated Ulex europaeus lectin, which is a fucose-binding lectin with a specificity for the H blood group antigen, showed that two of these glycosphingolipids carried this antigenic determinant . Preparations of crude and purified adhesin (a protein with a size of 15.7 kDa which lacked cysteine residues) from one of the strains also bound to these same two components . The third glycosphingolipid, which bound whole cells but neither preparation of adhesin, was recognized by Helix pomatia lectin, indicating that it contained N-acetylgalactosamine, possibly in the form of the A blood group antigen . Overlay assays with a sixth strain of C . albicans (GDH 2023) revealed a completely different binding pattern of four receptors, each of which contained N-acetylglucosamine . These results confirm earlier predictions about the receptor specificity of the strains made on the basis of adhesion inhibition studies and indicate that blood group antigens can act as epithelial cell receptors for C . albicans.

Infect Immun, 1996 Mar, 64(3), 751 - 5
Evidence of multiple extracellular phospholipase activities of Aspergillus fumigatus; Birch M et al.; Extracellular phospholipase activity has been implicated in the pathogenesis of several bacterial infections . Recently, extracellular phospholipase activity has been proposed as a virulence factor in the opportunistic yeast Candida albicans . Aspergillus fumigatus is the most pathogenic member of its genus, responsible for > 90% of infections . Previously, no specific virulence factors have been determined . We investigated the ability of A . fumigatus to produce extracellular phospholipases at 37 degrees C . Fast atom bombardment was used to compare lipid-containing media before and at 5-h intervals during shaking culture of A . fumigatus . Lipids were extracted and analyzed . Many anions corresponding to phospholipid breakdown products were identified . Specific anion species identified indicated phospholipase A, B, C (PLC), and D activities . PLC activity was further investigated by using the synthetic substrate p-nitrophenylphosphorylcholine . PLC activity was initially observed after 30 h of growth and accumulated in broth cultures up to 50 h . At 55 h, there was a sharp increase in PLC activity which coincided with cultures reaching the stationary phase . Activity of the PLC was measured at different temperatures, with greater activity occurring at 37 degrees C than at lower temperatures . Phospholipases could represent a virulence determinant in A . fumigatus.

Mil Med, 1996 Mar, 161(3), 143 - 5
Tincture of benzoin: clinical and microbiological implications of reusable containers; Wascher RA et al.; At our institution, tincture of benzoin solution is commonly used as a topical adhesive agent . As a cost-saving practice, multiple-dose bottles are routinely used in the operating rooms and the clinic on multiple patients . Although clinically pathogenic organisms are known to be capable of survival in both benzoin and its isopropyl alcohol solvent, no prior controlled studies have investigated the potential for tincture of benzoin solution to support the growth of specific pathogens under clinically relevant conditions . In this study, multiple aerobic, anaerobic, and spore-forming bacteria were exposed to tincture of benzoin solution, as well as Candida albicans and Mycobacterium fortuitum . Bacillus cereus was the only index organism demonstrating a clear ability to survive a 15 minute incubation in tincture of benzoin, although 24 hours of exposure to tincture of benzoin resulted in no subsequent viable cultures of this organism after 72 hours of incubation . Thus although certain bacilli might, under ideal circumstances, remain viable and infectious within multiple-dose bottles of tincture of benzoin, the risk of causing iatrogenic infection appears to be rather minimal . Still, the use of multiple-dose dispensers of topical agents, particularly in surgical patients, should be carefully scrutinized for their clinical risk-to-economic benefit ratio.

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