Z Naturforsch {C}, 2002 Mar-Apr, 57(3-4), 291 - 5 Antifungal garcinia acid esters from the fruits of Garcinia atroviridis; Mackeen MM et al.; Two new garcinia acid derivatives, 2-(butoxycarbonylmethyl)-3-butoxycarbonyl-2-hydroxy-3-propanolide and 1',1"-dibutyl methyl hydroxycitrate, were isolated from the fruits of Garcinia atroviridis guided by TLC bioautography against the fungus Cladosporium herbarum . The structures of these compounds were established by spectral analysis . The former compound represents a unique beta-lactone structure and the latter compound is most likely an artefact of garcinia acid (= hydroxycitric acid) . Both compounds showed selective antifungal activity comparable to that of cycloheximide (MID: 0.5 microg/spot) only against C herbarum at the MIDs of 0.4 and 0.8 microg/spot but were inactive against bacteria (Bacillus subtilis, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli), other fungi (Alternaria sp., Fusarium moniliforme and Aspergillus ochraceous) including the yeast Candida albicans. J Med Chem, 2002 Jun 20, 45(13), 2720 - 32 Antifungal agents . 10 . New derivatives of 1-{(aryl){4-aryl-1H-pyrrol-3-yl}methyl}-1H-imidazole, synthesis, anti-candida activity, and quantitative structure-analysis relationship studies; Tafi A et al.; The synthesis, anti-Candida activity, and quantitative structure-activity relationship (QSAR) studies of a series of 2,4-dichlorobenzylimidazole derivatives having a phenylpyrrole moiety (related to the antibiotic pyrrolnitrin) in the alpha-position are reported . A number of substituents on the phenyl ring, ranging from hydrophobic (tert-butyl, phenyl, or 1-pyrrolyl moiety) to basic (NH(2)), polar (CF(3), CN, SCH(3), NO(2)), or hydrogen bond donors and acceptor (OH) groups, were chosen to better understand the interaction of these compounds with cytochrome P450 14-alpha-lanosterol demethylase (P450(14DM)) . Finally, the triazole counterpart of one of the imidazole compounds was synthesized and tested to investigate influence of the heterocyclic ring on biological activity . The in vitro antifungal activities of the newly synthesized azoles 10p-v,x-c' were tested against Candida albicans and Candida spp . at pH 7.2 and pH 5.6 . A CoMFA model, previously derived for a series of antifungal agents belonging to chemically diverse families related to bifonazole, was applied to the new products . Because the results produced by this approach were not encouraging, Catalyst software was chosen to perform a new 3D-QSAR study . Catalyst was preferred this time because of the possibility of considering each compound as a collection of energetically reasonable conformations and of considering alternative stereoisomers . The pharmacophore model developed by Catalyst, named HYPO1, showed good performances in predicting the biological activity data, although it did not exhibit an unequivocal preference for one enantiomeric series of inhibitors relative to the other . One aromatic nitrogen with a lone pair in the ring plane (mapped by all of the considered compounds) and three aromatic ring features were recognized to have pharmacophoric relevance, whereas neither hydrogen bond acceptor nor hydrophobic features were found . These findings confirmed that the key interaction of azole antifungals with the demethylase enzyme is the coordination bond to the iron ion of the porphyrin system, while interactions with amino acids localized in proximity of heme could modulate the biological activity of diverse antifungal agents . In conclusion, HYPO1 conveys important information in an intuitive manner and can provide predictive capability for evaluating new compounds. J Biol Chem, 2002 Aug 23, 277(34), 30598 - 605 Epub 2002 Jun 11. Characterization and functional analysis of the siderophore-iron transporter CaArn1p in Candida albicans; Hu CJ et al.; Siderophores are small organic compounds with high affinity for ferric iron . Microorganisms commonly acquire iron via siderophore secretion and uptake . Here we report the characterization of the siderophore transporter CaArn1p in the fungal pathogen Candida albicans . Deletion of CaARN1 reduced the ability of C . albicans to use iron bound to the hydroxamate-type siderophore ferrichrome and abolished it when two high-affinity iron permease genes (CaFTR1 and CaFTR2) were also deleted, indicating a role of CaArn1p as well as the permeases in ferrichrome-iron uptake . Caarn1Delta (but not Caftr1DeltaCaftr2Delta) assimilated iron from another hydroxamate-type siderophore, ferrioxamine B, suggesting that iron uptake from this compound depends on the permeases, but not on CaArn1p . Northern blot analysis revealed that the transcription repressor CaTup1p repressed CaARN1 expression under iron-replete conditions via the DNA-binding protein Rfg1p . Green fluorescent protein-tagged CaArn1p was observed predominantly in the plasma membrane, with some in the cytoplasm as distinct spots . The number of these spots increased with the increase in ferrichrome concentration, suggesting that CaArn1p internalization might be a mechanism for ferrichrome-iron uptake or for recycling the transporter . Caarn1Delta did not show reduced virulence when injected into the blood stream of mice, implying that CaArn1p is not required for iron uptake along this route of infection. Int Endod J, 2002 Apr, 35(4), 321 - 9 Prevalence of yeasts in saliva and root canals of teeth associated with apical periodontitis; Egan MW et al.; AIMS: To determine: (i) the relative prevalence and diversity of yeasts in salivary and root canal samples from the same patients; and (ii) the clinical factors associated with their presence in saliva and root canals . METHODOLOGY: Sixty root canal samples from teeth associated apical periodontitis and the corresponding whole unstimulated saliva samples were obtained from 55 patients . The medical history including antibiotic therapy and clinical/radiographic data on the teeth were recorded . The samples were serially diluted and cultured on yeast & fungi-selective sabouraud dextrose agar . Isolates were characterized and speciated by the germ tube formation test, hyphal morphology and a commercial biochemical test kit (Rapid ID32C(R) system) . RESULTS: Twenty-three yeast isolates were recovered from 19 saliva samples and eight isolates from six root canal samples . Candida albicans (17/23 & 3/8) and Rodotorula mucilaginosa (2/23 & 4/8) were the most prevalent isolates from saliva and root canal samples . It was significantly (13.8 times) more probable that yeasts would be recovered from root canals when they were also present in the saliva (P = 0.021) . The effect of coronal restoration leakage (P = 0.08) and previous root canal treatment (P = 0.123) were equivocal . The history of antibiotic therapy had no association with the presence of yeasts in saliva (OR = 1.1) . CONCLUSIONS: Yeasts occurred relatively infrequently (10%) in root canals . Their presence in root canals was significantly associated with their presence in saliva . The role of yeasts in the initiation and perpetuation of periapical disease remains to be determined. Inflamm Res, 2002 Apr, 51(4), 213 - 7 Neutrophil hypochlorous acid production is impaired in multiple organ failure patients with candidaemia; reversal with antifungal agents; Hussien M et al.; OBJECTIVE AND DESIGN: Neutrophil-derived hypochlorous acid (PMN-HOCL) is a potent bactericidal and fungicidal agent . Neutrophils from patients with multiple organ failure (MOF) have an altered ability to inhibit Candida albicans . Conversely antifungal agents are effective in decreasing mortality in MOF patients with fungaemia . We determined the effect on the rate of production of PMN-HOCL in a series of MOF patients with C . albicans fungaemia when antifungal agents are administered . SUBJECTS AND METHODS: Eleven patients with MOF were recruited . Four patients had culture-positive C . albicans fungaemia (group 1) and 7 patients with MOF but without C . albicans fungaemia acted as controls (group 2) . HOCL assays were performed on peripheral blood PMN obtained from group 1 and group 2 patients . RESULTS: The maximum PMN-HOCL production rate for group 1 increased from 0.24 nmol/million PMN/min to 0.8 nmol/million PMN/min after treatment with antifungal agents (p < 0.01) (Mann-Whitney U test) whereas the PMN-HOCL kinetic (rate) curves for group 1 post-treatment and for group 2 were similar (student's-t-test) . CONCLUSIONS: These findings suggest that antifungal treatment improve PMN-HOCL production in MOF patients with C . albicans fungaemia and may improve the effectiveness in eradicating the organism. J Oral Sci, 2002 Mar, 44(1), 41 - 8 Effect of commercial ethanol propolis extract on the in vitro growth of Candida albicans collected from HIV-seropositive and HIV-seronegative Brazilian patients with oral candidiasis; Martins RS et al.; The present study assessed the susceptibility of Candida albicans strains, collected from HIV-positive patients with oral candidiasis, to a commercial 20% ethanol propolis extract (EPE) and compare it to the inhibitory action of the standardized antifungal agents nystatin (NYS), clotrimazole (CL), econazole (EC), and fluconazole (FL) . Twelve C . albicans strains collected from HIV-positive patients with oral candidiasis were tested . The inhibition zones were measured with a pachimeter and the results are reported as means and standard deviation (M +/- SD) . Data were analyzed statistically by the non-parametric Kruskal-Wallis test . EPE inhibited all the C . albicans strained tested . No significant difference was observed between the results obtained with NYS and EPE, while significant differences were observed between EPE and other antifungals . The C . albicans strains tested showed resistance to the remaining antifungal agents . The propolis extract used in this study inhibited the in vitro growth of C . albicans collected from HIV-seropositive Brazilian patients, creating/forming inhibition zones like those ones formed by NYS . This fact suggests that commercial EPE could be an alternative medicine in the treatment of candidiasis from HIV-positive patients . However, in vivo studies of the effect of EPE are needed to determine its possible effects on the oral mucosa. Med Mycol, 2002 Apr, 40(2), 139 - 42 Effects of erythromycin, clarithromycin, roxithromycin and azithromycin on murine gut colonization by Candida albicans; Samonis G et al.; Male Crl:CD1(ICR) BR mice were fed either chow containing Candida albicans or regular chow . The gastrointestinal tract of the C . albicans-fed mice was permanently colonized by the yeast . Groups of C . albicans-colonized mice were subsequently treated either with a macrolide (erythromycin, clarithromycin, roxithromycin or azithromycin) for 10 days or a normal saline solution (controls) . Other controls included non-colonized mice receiving the same antibiotics or a saline solution . Our data are as follows: (i) C . albicans-colonized mice treated with each macrolide had highly significant increase in colony counts of C . albicans in their stools compared to C . albicans-colonized mice treated with saline only; (ii) discontinuation of macrolide treatment showed a trend towards lower colony counts, which was not statistically significant (colony counts were sustained even after discontinuation of antibiotic treatment); (iii) dissemination of C . albicans did not occur; (iv) mice fed regular chow treated with the study drugs or saline did not have any yeasts in their stools . In conclusion, oral erythromycin, clarithromycin, roxithromycin and azithromycin cause a modest increase of the C . albicans concentration of the gastrointestinal tract . This increase is not associated with a higher risk of disseminated candidiasis. Med Mycol, 2002 Apr, 40(2), 123 - 9 Influence of sub-inhibitory concentrations of conventional antifungals on metabolism of Candida albicans and on its adherence to polystyrene and extracellular matrix proteins; Imbert C et al.; Five antifungal agents with different mechanisms of action were compared for their ability to affect mitochondrial dehydrogenase activity and adherence capacity of Candida albicans to polystyrene and extracellular matrix proteins . Only amphotericin B inhibited mitochondrial dehydrogenase activity when the culture medium was supplemented with galactose . 5-Fluorocytosine and terbinafine did not affect this activity, whereas itraconazole and fluconazole improved it . Furthermore, in these experimental conditions, the effect of sub-inhibitory concentrations of antifungals on adherence was dependent on the tested antifungal and the adherence surface: amphotericin B inhibited adherence to polystyrene and fibrinogen, but improved adherence to extracellular matrix . For all surfaces tested, when culture medium was supplemented with galactose, fluorocytosine did not affect adherence, and itraconazole, fluconazole and terbinafine inhibited adherence . Our results also confirmed the influence of the carbohydrates: sub-minimum inhibitory concentrations (MIC) of itraconazole increased or did not modify the mitochondrial metabolism of yeasts when the culture medium was supplemented with galactose, but this antifungal always decreased mitochondrial metabolism when the culture medium was supplemented with glucose . These data indicate that antifungals used below their MIC values can have various effects . It is important to distinguish the effects of antifungals on the metabolism of C . albicans from effects on its adherence capacity . The former effects are linked to the viability of the yeast and the latter depends on the colonization of cellular as opposed to inert surfaces. Folia Microbiol (Praha), 2002, 47(2), 179 - 81 Candida species isolated from cerebrospinal fluid; Dorko E et al.; In the period from November 1998 to June 2001 13 cases of nosocomial meningitis were reported . Candida albicans was isolated from 54% of the patients (7); C . parapsilosis from 23% (3); C . tropicalis from 15% (2) and C . krusei from 8% (1) . C . albicans was isolated from the cerebrospinal fluid (CSF) of five children with the following diagnoses: nonspecified tumor of the central nervous system, Hodgkin's disease, meningitis, suspect neuroinfection, and sepsis . Examination of CSF allowed us to detect 2 strains of C . albicans from adult patients, one after neurosurgery because of a brain tumor and one with a vascular disease of the brain . C . parapsilosis was found in CSF from two premature children and one child with epilepsy . Two isolates of C . tropicalis were obtained from both blood and the CSF of a child from the neonatal intensive care unit and from a child from pediatric oncology with multiple malignant neoplasms . Only one strain of C . krusei was found in the oral cavity and CSF of a patient after neurosurgery performed after head trauma. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 2000, 32(5), 509 - 515 Cloning and Identification of Genes Related with Morphogenesis of Candida albicans; Chen X et al.; A Candida albicans cDNA library was constructed and screened by differential hybridization . In hybridization using probes derived from population of yeast cells or hyphae, 67 recombinant phages exhibited more intense signal with the probe derived from hyphae than with the probe from yeast cells . One phage behaved vice versa . Physical map analysis and nucleotide sequence analysis suggested that cDNA of the CX1, a clone specific for yeast form, coded for cytochrome P450 L1A1 (Lanosterol 14alpha-demethylase) . Its specific expression pattern was confirmed by Northern analysis . Inhibition of serum on the expression of CX1 cDNA was observed . CX2 cDNA was one of those giving intensive signal with hyphae probes . The cDNA sequence contained a tandem repeat sequence, which was also found in ALS1, another Candida albicans gene identified, whose expression was related with morphogenesis . Northern analysis proved that it was expressed intensively with hyphae probes, however the expression could not be detected in those strongly hybridized to yeast cell probes . The locations of both cDNA on chromosome were analyzed. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 2000, 32(6), 586 - 594 Cloning and Functional Analysis of ALS Family Genes from Candida albicans; Chen X et al.; With a 0.5 kb probe of CX2, distribution of CX2 tandem repeats was studied in different C.albicans strains . Results suggest that all the C.albicans strains tested contained the tandem repeat . In order to verify if the expression of CX2 was hyphal specific, its expression was analyzed under various inductive and non-inductive conditions . With CX2 0.5 kb probe, Northern hybridization analysis confirmed that it was specifically in hyphae . The result of chromosomal localizationtion and genomic Southern blot analysis suggested that there were other genes containing the tandem repeat besides of ALS1 . A C.albicans s genomic DNA library was screened with the CX2 0.5 kb probe and several positive recombinant lambda phages were obtained . After endonuclease identification, subcloning, and sequence analysis, several ALS family genes were cloned . No.1 lambda phage DNA contained ALS4, No.4 lambda phage DNA contained ALS1, No.6 lambda phage DNA contained ALS3 . To study the role of ALS family genes in yeast-hyphal transition, als1/ALS1 mutant was constructed by homologous recombination . The ability to form hyphae was tested in different inductive culturing conditions . Defective hyphal growth were observed in some solid media. Microbiology, 2002 Jun, 148(Pt 6), 1737 - 46 Phosphatidylinositol-4-phosphate 5-kinase activity is stimulated during temperature-induced morphogenesis in Candida albicans; Hairfield ML et al.; Phosphoinositides are important lipid signalling molecules in eukaryotic cells . Phosphatidylinositol-4-phosphate 5-kinase (PI4P5K) catalyses the production of phosphatidylinositol 4,5-bisphosphate (PIP2), which stimulates phospholipase D1 (PLD1) activity in mammalian and yeast cells . PLD1 catalyses the formation of phosphatidic acid (PA), which has been shown to activate PI4P5Ks in mammalian and Saccharomyces cerevisiae cells . In the present study, PI4P5K activity in the opportunistic pathogen Candida albicans was identified . A gene with significant sequence homology to the S . cerevisiae PI4P5K was cloned and designated MSS4 . This gene was demonstrated to encode a functional PI4P5K by expression in S . cerevisiae . This enzyme was found to be membrane-associated and was stimulated by PA . Within the first 20 min after induction of polarized hyphal growth induced by a shift to elevated temperature, PI4P5K activity increased 2.5-fold . This stimulation was not observed when hyphae were induced by a combination of elevated temperature and serum . A lack of PLD1 activity resulted in the loss of induction of PI4P5K activity during the morphogenetic switch . Furthermore, the addition of propranolol attenuated the stimulation of PI4P5K activity during morphogenesis . These results suggest that PA derived from PLD1 activity stimulates C . albicans PI4P5K during the switch to the hyphal form under some conditions. Biochem Biophys Res Commun, 2002 Apr 26, 293(1), 231 - 8 Design of novel peptide analogs with potent fungicidal activity, based on PMAP-23 antimicrobial peptide isolated from porcine myeloid; Lee DG et al.; PMAP-23 is a 23-mer peptide derived from porcine myeloid . To develop novel antifungal peptides useful as therapeutic drugs, it would require a strong fungicidal activity against pathogenic fungal cells . To this goal, several analogs, with amino acid substitutions, were designed to increase the net hydrophobicity by Trp (W)-substitution at positions 10, 13, or 14 at the hydrophilic face of PMAP-23 without changing the hydrophobic helical face . The Trp (W)-substitution (P6) showed an enhanced fungicidal and antitumor activities, with the fungicidal activity inhibited by salts and the respiratory inhibitor, NaN(3) . The results suggested that the increase of hydrophobicity of the peptides correlated with fungicidal activity . The fungicidal effects of analog peptides were further investigated using 1,6-diphenyl-1,3,5-hexatriene (DPH) as a membrane probe . In Candida albicans, the analog peptide (P6) exerted its fungicidal effect on the blastoconidia in 20% fetal bovine serum by disrupting the mycelial forms . Furthermore, P6 caused significant morphological changes, and these facts suggested that the fungicidal function of the novel analog peptide (P6) was by damaging the fungal cell membranes . Thus, this peptide may provide a useful template for designing novel antifungal peptides useful for the treatment of infectious diseases. Med Mycol, 2001 Oct, 39(5), 401 - 8 Mechanism of antifungal action of kanosamine; Janiak AM et al.; The antibiotic kanosamine inhibited growth of Saccharomyces cerevisiae and a range of human pathogenic fungi, including Candida albicans . Kanosamine was transported into C . albicans cells by the glucose transport system and subsequently phosphorylated . The product of its intracellular metabolism, kanosamine-6-phosphate, was an inhibitor of the enzyme glucosamine-6-phosphate synthase . Inhibition was competitive in respect to one of the substrates, D-fructose-6-phosphate, with Ki = 5.9 mM, and was non-competitive in respect to the second substrate, L-glutamine . On the other hand, kanosamine-6-phosphate had no effect on the enzyme catalysing the next metabolic step, namely glucosamine-6-phosphate N-acetylase . The action of kanosamine on C . albicans cells resulted in profound morphological changes, inhibition of septum formation and cell agglutination . Experiments with S . cerevisiae mutants showed that the presence of the Cdr1p drug efflux pump did not affect the antifungal activity of kanosamine. Med Mycol, 2001 Oct, 39(5), 387 - 94 The Candida albicans cell wall-associated glyceraldehyde-3-phosphate dehydrogenase activity increases in response to starvation and temperature upshift; Gil ML et al.; We have determined the effect of environmental factors (mild thermal upshift and starvation) on the Candida albicans cell wall-associated glyceraldehyde-3-phosphate dehydrogenase (cwGAPDH) activity . Temperature upshift (from 28 to 37 degrees C) and/ or starvation (at 28 or 37 degrees C in water) of exponentially growing yeast cells caused an increase in cwGAPDH activity (3 to 5-, and 7 to 8-fold, respectively) . This increase in activity did not correlate with an increase in the amount of cwGAPDH protein present, as determined by flow cytometry, immunoelectron microscopy and Western-blotting . These results indicate that thermal upshift and starvation cause an activation of the cwGAPDH in C . albicans cells. Diagn Microbiol Infect Dis, 2002 May, 43(1), 19 - 28 Diversity of commensal yeasts within and among healthy hosts; Kam AP et al.; We sampled commensal yeasts from three body sites of 24 healthy individuals to examine the patterns of commensal yeast species distribution and strain relatedness within and among individuals . To examine the short-term dynamics, each individual was sampled three times every 35-40 days at each of three body sites: mouth, fingernail, and toenail . The hosts included six genealogically unrelated individuals and 18 that belonged to four families . A total of 63 morphologically distinct colonies were isolated, identified, and genotyped . Nine yeast species were recovered, including 28 isolates of Candida albicans; 26 of C . parapsilosis; 2 each of C . krusei and C . tropicalis; and 1 each of C . famata, C . glabrata, C . guilliermondii, C . lusitaniae and Trichosporon beigelii . A significant difference in total yeast recovery rate between families was observed . However, body sites did not differ in the rates of yeast recovery . The three body sites showed different species distributions with the fingernail sample containing the highest species diversity, followed by the toenail sample . The oral sample contained the lowest species diversity with all 23 oral isolates being C . albicans . Among the 63 strains, forty-six unique genotypes were identified by PCR fingerprinting . Eleven shared-genotypes were identified, seven of which were from the same body site of the same host . The other four were from different members of the same family . Several family-specific genotypes and genotype clusters were found but the results were inconsistent with strict familial transmission of human commensal yeasts . A single host can have multiple species or multiple genotypes of the same species at the same or different body sites . Changes of species and genotypes over the sampling period for the same body site of individual hosts were also observed, including one direct observation of familial yeast transmission between two members of the same family during our sampling period . Our results indicate dynamic processes of yeast colonization, maintenance and evolution in healthy human hosts. Curr Pharm Des, 2002, 8(13), 1137 - 54 Genomics strategies for antifungal drug discovery--from gene discovery to compound screening; Willins DA et al.; The use of genomics tools to discover new genes, to decipher pathways or to assign a function to a gene is just beginning to have an impact . Genomics approaches have been applied to both antibacterial and antifungal target discovery in order to identify a new generation of antibiotics . This review discusses genomics approaches for antifungal drug discovery, focusing on the areas of gene discovery, target validation, and compound screening . A variety of methods to identify fungal genes of interest are discussed, as well as methods for obtaining full-length sequences of these genes . One approach is well-suited to organisms having few introns (Candida albicans), and another for organisms with many introns (Aspergillus fumigatus) . To validate broad spectrum fungal targets, the yeast Saccharomyces cerevisiae was used as a model system to rapidly identify genes essential for growth and viability of the organism . Validated targets were then exploited for high-throughput compound screening. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 2001, 33(2), 198 - 204 Construction of Candida albicans Two-hybrid Library and Screening for Proteins Interacting with Crk1; Ni J et al.; In order to study interactions among proteins involved in Candida albicans morphorgenesis, C.albicans genomic DNA was digested with Sau3AI and fused to activating domain(AD) of LEXA to construct an AD-fused C.albicans genomic DNA expression library . The library contained 7.7x10(4) independent clones and included 1.2x10(5) kb DNA which was 10 fold of the C.albicans whole genomic DNA . The CRK1 gene encodes a CDC2-related protein, that was involved in morphorgenesis of C.albicans . The Crk1 fused to DNA binding domain of LexA was used as the bait to screen the LexA library . Among 9x10(5) transformants, eight Leu and LacZ double positive clones were selected . With restriction enzyme analysis, three kinds of fragments were identified . The clones NJ1, NJ2, NJ3 were sequenced and analyzed with BLAST program . It was found that NJ1, NJ2, NJ3 were homologs of S.cerevisiae gene STI1, RET2 and NFI1, respectively . The kinase domain of Crk1(Crk1N)interacted weakly with protein encoded by NJ1, while the noncatalytic domain of Crk1(Crk1C) interacted strongly with proteins encoded by NJ2 and NJ3 . The interaction of NJ3 and Crk1C was verified by the coimmunoprecipition of NJ3-HA with Crk1C-LexA . These proteins may be involved in maturation, transport, localization, and activity regulation of the Crk1. Wiad Lek, 2002, 55(1-2), 19 - 28 {Microflora of gastric juice in patients after eradication of Helicobacter pylori and treatment with a proton pump inhibitor}; Goscimski A et al.; Previous studies have suggested that decrease of acidity of gastric juice leads to microbial overgrowth in gastric juice . AIM OF THE STUDY: Qualitative and quantitative evaluation of upper respiratory tract and gastric juice microflora depending on the change of gastric juice pH, which occurred during treatment with pantoprazole and eradication therapy . MATERIAL AND METHODS: 40 patients suffering from duodenal ulcer or reflux esophagitis have been examined . Group 1, patients without infection of H . pylori have received pantoprazole 40 mg/d for 4 weeks . Group 2, patients with infection of H . pylori have received 7-day eradication therapy (pantoprazole, amoxicillin, clarithromycin) and then pantoprazole 40 mg/d for 3 weeks . The samples of gastric juice have been taken twice: before treatment and in the 4th week of treatment . The assessment of gastric juice included pH measurement and bacteriological analysis . RESULTS: The mean values of gastric juice pH in the group 1 were as following--1.7 (SD +/- 0.53) before and 5.2 (SD +/- 2.26) during the treatment . Mean microbial counts were: 0.47 (SD +/- 1.01) logCFU/ml before, and 1.77 (SD +/- 1.48) logCFU/ml during the treatment . In group 2 the increase of approximate amount of gastric juice from 1.8 (SD +/- 0.77) before treatment to 3.8 (SD +/- 2.43) during the treatment was noticed . The average titer of gastric juice microflora increased from 0.5 logCFU/ml (SD +/- 0.83) to 1.39 logCFU/ml (SD +/- 1.52) . Similarity between microflora isolated from gastric juice and tonsil swabs was noticed among most of the patients . CONCLUSIONS: The increase of gastric juice pH during the treatment with pantoprazole can lead to microflora growth in gastric juice . Microorganisms isolated from gastric juice among patients treated with antisecretive drugs mainly derived from the upper respiratory tract . Mostly isolated strains were: S . aureus, E . coli, Candida albicans. Glycobiology, 2002 Apr, 12(4), 251 - 60 Characterization of glucosylceramides in Pseudallescheria boydii and their involvement in fungal differentiation; Pinto MR et al.; Pseudallescheria boydii is a fungal pathogen that causes disease in immunocompromised patients . Ceramide monohexosides (CMHs) were purified from lipidic extracts of this fungus, showing that, as described for several other species, P . boydii synthesizes glucosylceramides as major neutral glycosphingolipids . CMHs from P . boydii were analyzed by high-performance thin-layer chromatography, gas chromatography coupled to mass spectrometry, fast atom bombardment-mass spectrometry, and nuclear magnetic resonance . These combination of techniques allowed the identification of CMHs from P . boydii as molecules containing a glucose residue attached to 9-methyl-4,8-sphingadienine in amidic linkage to 2-hydroxyoctadecanoic or 2-hydroxyhexadecanoic acids . Antibodies from a rabbit infected with P . boydii recognized CMHs from this fungus . Antibodies to CMH were purified from serum and used in indirect immunofluorescence, which revealed that CMHs are detectable on the surface of mycelial and pseudohyphal but not conidial forms of P . boydii, suggesting a differential expression of glucosylceramides according with morphological phase . We also investigated the influence of antibodies to CMH on growth and germ tube formation in P . boydii . Cultures that were supplemented with these antibodies failed to form mycelium, but the latter was not affected once formed . Similar experiments were performed to evaluate whether antibodies to CMH would influence germ tube formation in Candida albicans, a fungal pathogen that synthesizes glucosylceramide and uses differentiation as a virulence factor . Addition of antiglucosylceramide antibodies to cultures of C . albicans clearly inhibited the generation of germ tubes . These results indicated that fungal CMHs might be involved in the differentiation and, consequently, play a role on the infectivity of fungal cells. Br J Plast Surg, 2002 Apr, 55(3), 257 - 9 Candida colonisation within a silicone tissue expander; Saray A et al.; The fungal contamination of tissue expanders is rarely reported . There are, however, occasional reports of fungi in association with inflatable mammary implants . We describe the colonisation of a tissue expander with Candida albicans, resulting in the fluid becoming a turbid brown colour . The possible modes of inoculation and survival of the microorganism within the expander shell were investigated by means of biochemical and microbiological analyses of the fluid . The colonisation of silicone implants by opportunistic fungi is probably more common than has been reported, and precautions that can be taken to avoid this complication are emphasised . J Antimicrob Chemother, 2002 Jun, 49(6), 1007 - 10 Effect of matrix metalloprotease inhibitors on the 95 kDa metallopeptidase of Candida albicans; Imbert C et al.; A 95 kDa metallopeptidase of Candida albicans could be involved in the process of dissemination of the yeast . Matrix metalloproteases (MMPs) are also responsible for collagen breakdown in inflammatory and malignant processes . We tested six compounds on the C . albicans enzyme . Doxycycline, gentamicin, cefalothin, galardin, and elaidic and oleic acids are known for their capacity to inhibit some MMPs . Amongst these agents, only oleic acid was able to markedly inhibit the purified metallopeptidase at very low concentrations . Moreover, this fatty acid inhibited the secretion of the enzyme in the culture medium without altering the yeast viability. Dermatology, 2002, 204(3), 222 - 7 Euclidean and fractal computer-assisted corneofungimetry: a comparison of 2% ketoconazole and 1% terbinafine topical formulations; Arrese JE et al.; BACKGROUND: The corneofungimetry bioassay was designed as a unique model predicting the efficacy of topical and oral antifungals in dermatomycoses . OBJECTIVE: In this 2-step study performed in two groups of 15 volunteers, corneofungimetry was used to compare the effect of 5-day b.i.d . treatments with 2% ketoconazole and 1% terbinafine creams . METHODS: The bioassay was performed using 10 isolates of each of the 3 fungi Trichophyton rubrum, T . mentagrophytes var . interdigitale and Candida albicans put to grow on human stratum corneum . Controls were stratum corneum either untreated or enriched in propylene glycol contained in an unmedicated vehicle . Quantitative assessments were made using both Euclidean and fractal geometry parameters . RESULTS: In comparison with untreated stratum corneum, the fungitoxic activity of the 2% ketoconazole and 1% terbinafine formulations was obvious and similar against dermatophytes . By contrast, 2% ketoconazole was significantly more active against C . albicans than 1% terbinafine . The propylene-glycol-containing vehicle did not exhibit a significant effect upon the dermatophyte growth . Positive linear correlations were yielded between the extent area and the fractal dimension D of dermatophyte mycelia . By contrast, D appeared unrelated to the relative area of dermatophyte growth compared to controls . CONCLUSION: The combination of Euclidean and fractal analyses improves the information provided by the corneofungimetry bioassay . Creams containing 2% ketoconazole and 1% terbinafine appear equally effective against dermatophytes while the former is more potent against Candida albicans . J Clin Microbiol, 2002 Jun, 40(6), 2228 - 30 In vitro activity of caspofungin (MK-0991) against Candida albicans clinical isolates displaying different mechanisms of azole resistance; Bachmann SP et al.; Caspofungin inhibits the synthesis of 1,3-beta-D-glucan, a key step in fungal cell wall biosynthesis . Here we report on its potent in vitro activity (MIC at which 90% of the isolates tested are inhibited = 1 microg per ml of RPMI medium) against 32 Candida albicans fluconazole-susceptible and -resistant clinical isolates irrespective of the underlying resistance mechanism (alterations in ERG11 and/or upregulation of MDR and CDR genes encoding efflux pumps) and provide further evidence that caspofungin is not a substrate for multidrug transporters. J Clin Microbiol, 2002 Jun, 40(6), 2182 - 6 Fluorescence in situ hybridization with peptide nucleic acid probes for rapid identification of Candida albicans directly from blood culture bottles; Rigby S et al.; A new fluorescence in situ hybridization (FISH) method that uses peptide nucleic acid (PNA) probes for identification of Candida albicans directly from positive-blood-culture bottles in which yeast was observed by Gram staining (herein referred to as yeast-positive blood culture bottles) is described . The test (the C . albicans PNA FISH method) is based on a fluorescein-labeled PNA probe that targets C . albicans 26S rRNA . The PNA probe is added to smears made directly from the contents of the blood culture bottle and hybridized for 90 min at 55 degrees C . Unhybridized PNA probe is removed by washing of the mixture (30 min), and the smears are examined by fluorescence microscopy . The specificity of the method was confirmed with 23 reference strains representing phylogenetically related yeast species and 148 clinical isolates covering the clinically most significant yeast species, including C . albicans (n = 72), C . dubliniensis (n = 58), C . glabrata (n = 5), C . krusei (n = 2), C . parapsilosis (n = 4), and C . tropicalis (n = 3) . The performance of the C . albicans PNA FISH method as a diagnostic test was evaluated with 33 routine and 25 simulated yeast-positive blood culture bottles and showed 100% sensitivity and 100% specificity . It is concluded that this 2.5-h method for the definitive identification of C . albicans directly from yeast-positive blood culture bottles provides important information for optimal antifungal therapy and patient management. Phytochemistry, 2002 Jun, 60(4), 323 - 7 Triterpene glycosides of Lupinus angustifolius; Woldemichael GM et al.; Investigation of whole seeds of Lupinus angustifolius L . (Leguminosae) yielded the two triterpenoid saponins with branched monosaccharide chain 3 beta,21 beta,22 beta,24-tetrahydroxyolean-12-en-3-O-alpha-L-rhamnopyranosyl-(1-->3)-{alpha-L-rhamnopyranosyl-(1-->2)}-beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranoside (3) and 3 beta,21 beta,22 beta,24-tetrahydroxyolean-12-en-3-O-alpha-L-rhamnopyranosyl-(1-->3)-{alpha-L-rhamnopyranosyl-(1-->2)}-beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-21-O-alpha-L-rhamnopyranoside (4) along with the known compounds soyasaponin I (1) and 3 beta,21 beta,22 beta,24-tetrahydroxyolean-12-en-3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-21-O-alpha-L-rhamnopyranoside (2) . The structures of the compounds were elucidated using hydrolysis, FAB-MS and extensive NMR experiments . Compounds 2-4 showed moderate antifungal activity against Candida albicans with MIC values of 25, 25 and 30 microg/ml, respectively . Only soyasaponin I was found weakly hemolytic (HC(50) >500 microg/ml). Can J Microbiol, 2002 Apr, 48(4), 369 - 73 Utility of a pre-optimized kit for random amplified polymorphic DNA in typing Candida albicans; Riederer KM et al.; The utility of a pre-optimized kit for random amplified polymorphic DNA (RAPD) was assessed in typing diverse strains of Candida albicans from epidemiologically unrelated inpatients (interpatient analysis) and in detecting clonal variations that maybe present within individual patient isolates (intrapatient analysis) . Stool samples from inpatients were cultured on Inhibitory Mold agar . Nine individual colonies from all patients with > or =9 colonies of C . albicans (n = 18) were selected, frozen, and karyotyped using CHEF genomic DNA plug kits and CHEF-DRIII . Each of the selected colonies was then analyzed by RAPD, utilizing the selected kit, with 6 primers . Interpatient analysis revealed 9 karyotypes and 17 RAPD composites . RAPD discrimination was significantly better (p < 0.001) . Intrapatient analysis revealed 34 (21%) and 33 (20.4%) variants among 162 colonies tested by RAPD and karyotyping, respectively . The results were discordant in 25 variants, all with differences of 1-3 bands . These results illustrate that this pre-optimized kit for RAPD provides excellent discrimination of genetically unrelated strains . Its performance in delineating subtle clonal differences was comparable with karyotyping; both methods failed to detect all minor genetic variations . The ease of use and quick turnaround time of this kit offer a practical and reliable method for typing diverse strains of C . albicans, but may be inadequate for assessing microevolution. Scand J Infect Dis, 2002, 34(3), 197 - 200 Post-antifungal effect and effects of sub-MIC concentrations on previously treated Candida spp.: influence of exposure time and concentration; Garcia MT et al.; This study evaluates the influence of exposure time and concentration on the post-antifungal effect (PAFE) and the effect of sub-MIC concentrations (1/4 x MIC) on Candida albicans and C . glabrata in the PAFE stage (PAFSE) . This stage was induced by pretreatment for 1.5, 3 or 12 h with 1 x, 4 x or 8 x MIC of 4 antifungal agents fundamental to modern candidiasis therapy . The length of the 2 effects studied was dependent on the concentration of the antifungal agent applied during pretreatment, as well as on the exposure time . An increase in the dose and/or longer pretreatment prolonged the duration of the PAFE and PAFSE in both species and with all the antifungal agents . Significant PAFEs were always observed for amphotericin B and 5-fluorocytosine (0.8-13 h and 0.6-10.8 h, respectively) . These values were increased (by 2.3-8.7 h and 1.5-7.8 h, respectively) by posterior exposure to 1/4 x MIC of the respective antifungal agent . Neither ketoconazole nor fluconazole were able to induce significant PAFEs, even with exposures of up to 12 h duration and a dose of 8 x MIC . However, treatment with 1/4 x MIC of each of the 2 azoles led to significant PAFSEs in both yeast species, of up to 6.5 h duration with ketoconazole and 1.7 h with fluconazole, if the concentrations and/or exposure times were sufficiently high. Kansenshogaku Zasshi, 2002 Apr, 76(4), 231 - 7 {Antibiotic activity of P . aeruginosa against MRSA and Candida albicans}; Kondo S et al.; The antibiotic activity demonstrated by P . aeruginosa (Bacillus pyocyaneus) has been reported more than one hundred years ago by Emmerich et al (1899) . Studies on such bacterial interference between P . aeruginosa and other pathogenic bacteria or fungi have not been extensively reported in recent years . In this paper, we report on the anti MRSA activity and anti Candida activity demonstrated by clinical isolates of P . aeruginosa (34 strains) . The antibiotic activity was tested by reversed agar plate method, as previously reported, and the degree of the activity was expressed as the diameter of the zone of growth inhibition . The stability of both anti MRSA activity and anti Candida activity was evaluated at the time after 24 and 48-hr incubation . Also the effect of agar plate with or without 5% sheep blood on antibiotic activity was evaluated . Strong anti MRSA activity and anti Candida activity was shown at the time after 24-hr incubation . At the time after 48-hr incubation, anti MRSA activities were significantly suppressed but anti Candida activities were persisted . The inhibitory activity was correlated with dye production of P . aeruginosa . Some strains having non or weak dye production, showed the inhibitory activity by 48-hr incubation . Result from these strains without suppression of anti Candida activity by additional blood may suggest that the existence of a new factor produced by P . aeruginosa . Because of frequent isolation of MRSA or Candida from clinical materials, we must consider bacterial flora and bacterial interference against pathogenic bacteria at the time of the antibiotic choice for the patients infected or colonized with P . aeruginosa. Ann Hematol, 2002 May, 81(5), 233 - 43 Epub 2002 May 04. Invasive fungal infections in hematology: new trends; Martino R et al.; Invasive fungal infections (IFI) are among the most feared complications of patients being treated for a hematological malignancy . Currently, most serious IFI occur in patients with acute leukemia and after allogeneic hematopoietic stem cell transplantation . Although Candida albicans and Aspergillusspp . continue to be the main pathogens, the proportion of patients infected by non-albicans species of Candida and other yeasts and by other filamentous fungi is rising in most institutions . Risk factors for the various IFI differ, and it is thus of utmost importance to realize that not all patients are the same with respect to the risk for developing the various IFI . Recent advances in diagnosis now allow the use of very sensitive imaging techniques with an extremely low negative predictive value . Among the novel microbiologic methods, the galactomannan antigen test is now commercially available for routine use in the diagnosis of aspergillosis, while DNA fungal detection is still experimental . For the first time, clinicians now have a broad range of antifungals to chose from, with special emphasis on amphotericin B preparations, novel broad-spectrum azoles, and the echinocandins . However, the exact place of these agents in treating different IFI will need to be found in the near future. J Oral Rehabil, 2002 May, 29(5), 452 - 7 Mycological and cytological examination of oral candidal carriage in diabetic patients and non-diabetic control subjects: thorough analysis of local aetiologic and systemic factors; Kadir T et al.; In this study, 55 diabetic patients and 45 non-diabetic control subjects were examined to determine oral candidal carriage state . The influence of some local aetiologic and systemic factors such as: salivary flow rate and pH, heredity, alcohol drinking, smoking habits, antimicrobial therapy, wearing of denture, burning sensation, dry mouth, taste alteration and tooth brushing habit on candidal carriage rate were investigated . Imprint culture, cytological smears and biochemical tests were used . Oral carrier rate and density of Candida species were non-significantly higher in the diabetic patients than in the non-diabetic control subjects . This increase was confirmed cytologically too . In both groups, Candida albicans was found to be a predominant species on tongue dorsum . Cigarette and alcohol habits of men were higher while tooth brushing habit was less than in women in diabetic and control groups . Salivary flow rate and pH values of diabetic patients were significantly lower while serum glucose values were significantly higher than of non-diabetic controls . The rate of diabetic patients suffering from dry mouth and having diabetic heredity in the family were significantly higher than control subjects . The candidal colonization was higher and keratinization was lower while diabetic treatment tended from diet and oral antidiabetic towards insulin . The decrease in salivary pH, the increase in serum glucose and wearing denture were correlated with the increased rate and density of C . albicans in both groups . Keratinization was also accompanied with the increase in leucocytes . In diabetic group, positive correlations were found between antimicrobial therapy and C . glabrata carriage; the increase in leucocytes and C . albicans carriage; the increase in keratinization and alcohol habit; serum glucose and smoking habit; dry mouth complaint and antimicrobial therapy . There was a negative correlation between salivary flow rate and C . albicans carriage . In control group a positive correlation was found between antimicrobial therapy and keratinization. Lett Appl Microbiol, 2002, 34(6), 398 - 401 Detection and quantification of phytotoxic metabolites of Sarocladium oryzae in sheath rot-infected grains of rice; Ghosh MK et al.; AIMS: The present study describes the detection and quantification of the Sarocladium oryzae metabolites, helvolic acid and cerulenin in extracts of rice grains collected from plants infected with sheath rot . It also describes the phytotoxicity of these metabolites on rice seedlings . METHODS AND RESULTS: Helvolic acid and cerulenin in sheath rot-infected rice grains were detected using thin layer chromatography (TLC) and nuclear magnetic resonance (NMR) analyses . On the TLC plates helvolic acid and cerulenin moved as brownish yellow spots and showed R(F) values of 0.61 and 0.49, respectively . A standard assay curve was developed on the basis of selective toxicity of helvolic acid towards Calvibacter michiganensis ATCC 2140 and cerulenin towards Candida albicans 1150 . The amounts of helvolic acid and cerulenin on the basis of standard assay curve were 2.2 and 1.75 microg g(-1) of infected seeds . Treatment of IR 36 rice seedlings with metabolites induced chlorosis and reduced shoot length by 20%, root length by 30% and root number by 7% relative to control . CONCLUSIONS: Helvolic acid and cerulenin were detected in infected rice grains and these metabolites induced chlorosis and reduced the seed viability and seedling health of rice . SIGNIFICANCE AND IMPACT OF THE STUDY: Antimicrobial and phytotoxic metabolites, helvolic acid and cerulenin are present in infected grains and reduce the seed viability and seedling health . These metabolites may increase the pathogenic potential and survival of S . oryzae in rice seed by competing with other seed-borne fungi. Mol Microbiol, 2002 Jun, 44(5), 1351 - 66 Host versus in vitro signals and intrastrain allelic differences in the expression of a Candida albicans virulence gene; Staib P et al.; The yeast Candida albicans is a harmless colonizer of mucosal surfaces in healthy people but can become a serious pathogen in immunocompromised patients, causing superficial as well as systemic infections . The evolution of gene families encoding pathogenicity-related functions, like adhesins and secreted aspartic proteinases (Saps), which are differentially induced by host signals at various stages of colonization and infection, may have allowed C . albicans an optimal adaptation to many different host niches . We found that even the two alleles of a single gene can be differentially regulated in the diploid C . albicans . In the model strain SC5314, the in vitro expression of one of the two SAP2 alleles, SAP2-1, depended on the presence of a functional SAP2-2 allele . In contrast, inactivation of SAP2-1 did not in-fluence the expression of SAP2-2 . The proteinase encoded by the SAP2-2 allele serves as a signal sensor and amplifier to enhance its own expression as well as to induce the SAP2-1 allele to achieve maximal proteolytic activity under appropriate conditions . Using in vivo expression technology, we could demonstrate that the SAP2-1 allele is significantly activated only in the late stages of systemic candidiasis in mice, whereas the SAP2-2 allele is induced much earlier . The differential regulation of the two SAP2 alleles was due to differences in their pro-moters, which contained a variable number of two pentameric nucleotide repeats . Mutations that reduced or increased the copy number of these repeats diminished the inducibility of the SAP2 promoter during infection but not in vitro, suggesting that the mutations affected interactions of regulatory factors that are necessary for SAP2 activation in vivo but dispensable for its induction in vitro . Therefore, the signals and signal transduction pathways that mediate SAP2 expression within certain host niches may differ from those that activate the gene in vitro . In addition to the generation of gene families whose members exhibit functional and regulatory diversification, C . albicans seems to use its diploid genome to create further variability and host adaptation by differential evolution of even the two alleles of a single gene. Infect Control Hosp Epidemiol, 2002 May, 23(5), 281 - 4 Genotypic analysis by 27A DNA fingerprinting of Candida albicans strains isolated during an outbreak in a neonatal intensive care unit; Boccia S et al.; We describe an outbreak of Candida albicans systemic infection involving five premature infants in a neonatal intensive care unit . Molecular and epidemiologic characterization of all C . albicans isolates was performed by DNA fingerprinting with the 27A probe . This genotypic analysis demonstrated that the isolates were identical, providing evidence for the circulation of a unique C . albicans strain. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 2002 May, 34(3), 298 - 304 {CaSRB9, a novel Candida albicans gene, plays a role in morphogenesis of Saccharomyces cerevisiae}; Zhou Z et al.; Candida albicans is the most frequently isolated fungal pathogen in humans . Many factors are involved in its morphological transition . Flo8 plays an important role in morphogenesis of Saccharomyces cerevisiae . In this work, a C.albicans genomic DNA library was introduced into an S.cerevisiae flo8/flo8 mutant to screen genes which could complement its invasive growth defect . In this screening, a novel gene was isolated and designated CaSRB9 (Candida albicans SRB9 gene).The CaSRB9 gene had an ORF of 4 998 bp, encoding a putative protein of 1 665 amino acids . The CaSrb9 shared highest similarity in amino acids (38%) with Srb9 of S.cerevisiae . Ectopic expression of the CaSRB9 gene in diploid S . cerevisiae suppressed defect in filamentous growth of some mutants in filamentation MAPK pathway (ste7/ste7, ste 12 / ste 12, and tec 1 / tec 1) and flo 8 / flo 8 mutant under nitrogen starvation conditions . In haploid S . cerevisiae, ectopic expressed CaSrb9 complemented the invasive growth defect of flo8 mutant but failed to complement the invasive growth defects of the mutants in filamentation MAPK pathway. Surgery, 2002 May, 131(5), 534 - 40 Enteral fluconazole is well absorbed in critically ill surgical patients; Pelz RK et al.; BACKGROUND: Enteral fluconazole, a triazole antifungal agent with an excellent oral bioavailability, has not been widely studied in critically ill surgical patients . METHODS: During a randomized placebo-controlled trial of enteral fluconazole (N = 130) versus placebo (N = 130) for the prevention of fungal infections in critically ill surgical patients, trough fluconazole levels were measured after the loading dose and 3 times weekly during intensive care unit stay . Minimum inhibitory concentrations (MICs) for fluconazole were measured on all infecting Candida isolates . RESULTS: Four hundred sixty-seven serum samples were assayed for fluconazole levels in 121 patients . The most common infecting fungal species was Candida albicans, isolated in 14 of 31 infections (45%) . Other infecting species were C glabrata, C tropicalis, and C parapsilosis . Mean fluconazole levels were above the highest MIC for C albicans and C parapsilosis in all but 5 patients (4%) . Mean fluconazole levels were below the median MIC for C glabrata in 93 of 121 patients (77%) . No significant relationship was seen between fluconazole levels and risk for fungal infection . CONCLUSIONS: Serum fluconazole levels are above the MIC of most yeast species found in these patients . These levels may not be above the MIC of C glabrata, the second most common Candida isolate causing infection in this study. Antimicrob Agents Chemother, 2002 Jun, 46(6), 1857 - 69 Comparative antifungal activities and plasma pharmacokinetics of micafungin (FK463) against disseminated candidiasis and invasive pulmonary aspergillosis in persistently neutropenic rabbits; Petraitis V et al.; Micafungin (FK463) is an echinocandin that demonstrates potent in vitro antifungal activities against Candida and Aspergillus species . However, little is known about its comparative antifungal activities in persistently neutropenic hosts . We therefore investigated the plasma micafungin pharmacokinetics and antifungal activities of micafungin against experimental disseminated candidiasis and invasive pulmonary aspergillosis in persistently neutropenic rabbits . The groups with disseminated candidiasis studied consisted of untreated controls (UCs); rabbits treated with desoxycholate amphotericin B (DAMB) at 1 mg/kg of body weight/day; or rabbits treated with micafungin at 0.25, 0.5, 1, and 2 mg/kg/day intravenously . Compared with the UCs, rabbits treated with micafungin or DAMB showed significant dosage-dependent clearance of Candida albicans from the liver, spleen, kidney, brain, eye, lung, and vena cava . These in vivo findings correlated with the results of in vitro time-kill assays that demonstrated that micafungin has concentration-dependent fungicidal activity . The groups with invasive pulmonary aspergillosis studied consisted of UCs; rabbits treated with DAMB; rabbits treated with liposomal amphotericin B (LAMB) at 5 mg/kg/day; and rabbits treated with micafungin at 0.5, 1, and 2 mg/kg/day . In comparison to the significant micafungin dosage-dependent reduction of the residual burden (in log CFU per gram) of C . albicans in tissue, micafungin-treated rabbits with invasive pulmonary aspergillosis had no reduction in the concentration of Aspergillus fumigatus in tissue . DAMB and LAMB significantly reduced the burdens of C . albicans and A . fumigatus in tissues (P < 0.01) . Persistent galactomannan antigenemia in micafungin-treated rabbits correlated with the presence of an elevated burden of A . fumigatus in pulmonary tissue . By comparison, DAMB- and LAMB-treated animals had significantly reduced circulating galactomannan antigen levels . Despite a lack of clearance of A . fumigatus from the lungs, there was a significant improvement in the rate of survival (P < 0.001) and a reduction in the level of pulmonary infarction (P < 0.05) in micafungin-treated rabbits . In summary, micafungin demonstrated concentration-dependent and dosage-dependent clearance of C . albicans from persistently neutropenic rabbits with disseminated candidiasis but not of A . fumigatus from persistently neutropenic rabbits with invasive pulmonary aspergillosis. Antimicrob Agents Chemother, 2002 Jun, 46(6), 1723 - 7 In vitro activities of ravuconazole and voriconazole compared with those of four approved systemic antifungal agents against 6,970 clinical isolates of Candida spp; Pfaller MA et al.; The in vitro activities of ravuconazole and voriconazole were compared with those of amphotericin B, flucytosine (5FC), itraconazole, and fluconazole against 6,970 isolates of Candida spp . obtained from over 200 medical centers worldwide . Both ravuconazole and voriconazole were very active against all Candida spp . (MIC at which 90% of the isolates tested are inhibited {MIC(90)}, 0.25 microg/ml; 98% of MICs were < or 1 microg/ml); however, a decrease in the activities of both of these agents was noted among isolates that were susceptible-dose dependent (fluconazole MIC, 16 to 32 microg/ml) and resistant (MIC, > or = 64 microg/ml) to fluconazole . Candida albicans was the most susceptible species (MIC(90) of both ravuconazole and voriconazole, 0.03 microg/ml), and C . glabrata was the least susceptible species (MIC(90), 1 to 2 microg/ml) . Ravuconazole and voriconazole were each more active in vitro than amphotericin B, 5FC, itraconazole, and fluconazole against all Candida spp . and were the only agents with good in vitro activity against C . krusei . These results provide further evidence for the spectrum and potency of ravuconazole and voriconazole against a large and geographically diverse collection of Candida spp. Antimicrob Agents Chemother, 2002 Jun, 46(6), 1704 - 13 Resistance mechanisms in clinical isolates of Candida albicans; White TC et al.; Resistance to azole antifungals continues to be a significant problem in the common fungal pathogen Candida albicans . Many of the molecular mechanisms of resistance have been defined with matched sets of susceptible and resistant clinical isolates from the same strain . Mechanisms that have been identified include alterations in the gene encoding the target enzyme ERG11 or overexpression of efflux pump genes including CDR1, CDR2, and MDR1 . In the present study, a collection of unmatched clinical isolates of C . albicans was analyzed for the known molecular mechanisms of resistance by standard methods . The collection was assembled so that approximately half of the isolates were resistant to azole drugs . Extensive cross-resistance was observed for fluconazole, clotrimazole, itraconazole, and ketoconazole . Northern blotting analyses indicated that overexpression of CDR1 and CDR2 correlates with resistance, suggesting that the two genes may be coregulated . MDR1 overexpression was observed infrequently in some resistant isolates . Overexpression of FLU1, an efflux pump gene related to MDR1, did not correlate with resistance, nor did overexpression of ERG11 . Limited analysis of the ERG11 gene sequence identified several point mutations in resistant isolates; these mutations have been described previously . Two of the most common point mutations in ERG11 associated with resistance, D116E and E266D, were tested by restriction fragment length polymorphism analysis of the isolates from this collection . The results indicated that the two mutations occur frequently in different isolates of C . albicans and are not reliably associated with resistance . These analyses emphasize the diversity of mechanisms that result in a phenotype of azole resistance . They suggest that the resistance mechanisms identified in matched sets of susceptible and resistant isolates are not sufficient to explain resistance in a collection of unmatched clinical isolates and that additional mechanisms have yet to be discovered. Antimicrob Agents Chemother, 2002 Jun, 46(6), 1688 - 94 Identification of major glucan-associated cell wall proteins of Candida albicans and their role in fluconazole resistance; Angiolella L et al.; Identification of major glucan-associated proteins (GAPs) of the cell wall of a number of Candida albicans isolates susceptible or resistant to fluconazole (FLC) was addressed by direct sequencing of the protein bands resolved by unidimensional gel electrophoresis . Changes in the GAP compositions of the different strains grown in the presence of the drug were also investigated . In the FLC-susceptible strains, the major (more abundant) GAPs were enolase (46 kDa), two isoforms of phosphoglyceromutase (32 and 29 kDa), and two beta-(1-3)-exoglucanases (44 and 34 kDa), one of which (the 34-kDa component) was glycosylated . When these strains were grown in the presence of FLC there were substantial decreases in the intensities of the two enzymes of the glycolytic pathway (enolase and the phosphoglyceromutases), which were apparently replaced by enhancement of the exoglucanase constituents, particularly the 44-kDa one . This GAP pattern closely mimicked that observed in the FLC-resistant strains whether they were grown in the presence or in the absence of the drug . Both the enolase and the exoglucanase constituents were detected in the culture supernatants of FLC-treated cells, together with substantial amounts of highly glycosylated, probably mannoprotein secretory material, suggesting that FLC may cause marked alterations of GAP incorporation into the cell wall . Altogether, we were able to identify all major GAP constituents and monitor their distributions in the cell wall of C . albicans during treatment with FLC . The near equivalence of the GAP profile for the FLC-susceptible strain grown in the presence of FLC to that for the FLC-resistant strain suggests that the effects of the drug on GAPs may be stably incorporated into the cell wall of the fungus upon acquisition of resistance. Antimicrob Agents Chemother, 2002 Jun, 46(6), 1634 - 9 Internal thiols and reactive oxygen species in candidacidal activity exerted by an N-terminal peptide of human lactoferrin; Lupetti A et al.; We previously showed that the energized mitochondrion and extracellular ATP are essential for the candidacidal activity of the N-terminal peptide of human lactoferrin, subsequently referred to as hLF(1-11) . The present study focuses on the involvement of internal thiols and reactive oxygen species (ROS) in the candidacidal activity exerted by hLF(1-11) . Our results reveal that hLF(1-11) reduced the internal thiol level of Candida albicans by 20% . In agreement, N-acetyl-L-cysteine (NAC), which is a precursor of glutathione and an ROS scavenger, inhibited the candidacidal activity of hLF(1-11) . In addition, azodicarboxylic acid bis(N,N-dimethylamide) (diamide), which oxidizes internal thiols, was candidacidal . Furthermore, hLF(1-11) increased the level of ROS production by C . albicans in a dose-dependent manner, and a correlation between ROS production and candidacidal activity was found . 6-Hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), which is an ROS scavenger, partially inhibited the hLF(1-11)-induced, but not the diamide-triggered, candidacidal activity . It is of interest that hLF(1-11) and diamide acted synergistically in killing C . albicans and in ROS production . In agreement, oxidized ATP, an irreversible inhibitor of extracellular ATP receptors, partially blocked the hLF(1-11)-induced, but not the diamide-triggered, candidacidal activity . Finally, the hLF(1-11)-induced activation of mitochondria was inhibited by NAC, indicating that internal thiols and ROS affect mitochondrial activity . Therefore, the candidacidal activity of hLF(1-11) involves both generation of ROS and reduction of internal thiols. Ginecol Obstet Mex, 2002 Feb, 70, 59 - 65 {Efficacy and tolerance of 200 mg of fenticonazole versus 400 mg of miconazole in the intravaginal treatment of mycotic vulvovaginitis}; Munoz Reyes JR et al.; OBJECTIVE: Fenticonazole is an imidazole derivative with a broad-spectrum antifungal activity mainly against Candida albicans . Fenticonazole nitrate was compared to myconazole vs . test efficacy, tolerance and treatment compliance in-patients with mycotic vulvovaginitis . PATIENTS AND METHOD: Eighty outpatients with mycotic vulvovaginitis were included and randomly placed in two groups of forty patients each one . Each group was either given 200 mg of fenticonazole nitrate intravaginal ovules or 400 mg of myconazole vaginal ovules on a daily basis for three days . Before the pharmacological treatment, each patient was evaluated clinically and microbiologically, as well as at the end of treatment (day 7-10) and one-month (28 days) after treatment . All patients gave informed consent to participate in the trial . RESULTS: No statistical differences were found between either treatment group during the anthropometric and demographic evaluation, neither in the signs and clinical symptoms of infection . Both treatment groups were considered as comparable . At after treatment evaluation (7-10 days) it was observed disappearance of almost all symptoms . At the final evaluation this fact was even more evident (day 21-28) . At the study's completion, clinical efficacy (symptoms relief) was considered satisfactory in 100% (40/40) of the cases in the fenticonazole group and 97.5% (39/40) of myconazole group (one way ANOVA {GLM} analysis) . The microorganisms more frequently isolated as the causing agents of infection in both groups were Candida albicans, 93% in the fenticonazole group and 85% in the myconazole group . Upon the study's completion, the microbiological efficacy was considered as successful in 97.5% of the cases in both groups (chi square {x2} analysis) . The patient's compliance to the treatment was considered satisfactory in 100% in the fenticonazole group and 97.5% in the myconazole group . Tolerance was considered excellent in 100% (40/40) of the fenticonazole cases and in 95% (38/40) of the myconazole cases . Nevertheless, 5% (2/40) of the patients in the myconazole group had minor adverse events that did not require treatment suspension . No adverse events were reported in the fenticonazole group . CONCLUSION: The results showed that both drugs are equally efficient in treating signs and symptoms of the infection as well as equally effective in the microbiological elimination of the causing agents of the mycotic vulvovaginitis. Yao Xue Xue Bao, 1998 Jun, 33(6), 429 - 35 {Synthesis and anti-Candida albicans properties of L-4-oxalysine-N3-4-methoxyfumaroyl-L-2,3-diaminopropanoic-containing peptide analogues}; Feng S et al.; In order to improve the inhibitory activity of L-4-oxalysine(abbreviated as I-677) against clinically important pathogen Candida albicans, double-warheads peptide analogues containing I-677 and N3-4-methoxyfumaroyl-L-2,3-diaminopropanoic acid(FMDP) were designed based on the concept of "Illicit Transport" and peptide transport specificities of C . albicans . One compound of I-677-FMDP and seven compounds of I-677-AA-FMDP(AA = Nva, Leu, Val, Phe, Pro, D-Pgly, D, L-p-Cl-Phe) were synthesized and examined for antifungal activities . The results of anti-Candida albicans test in vitro of these double- warheads peptide analogues containing I-677 and FMDP showed very high activities against Candida albicans . The molar MIC (molar minimum inhibitory concentration) ratio of free I-677 to I-677-AA-FMDP is 40-770 and that of FMDP is 60-1130 . The values of molar MIC of I-677-AA-FMDP varied from 6.56 x 10(-9) mol.disk-1 to 3.5 x 10(-10) mol.disk-1 . The results of competitive antagonism studies indicated that I-677-AA-FMDP were transported into Candida albicans cells by the di-tripeptide permease and subsequently hydrolyzed by intracellular peptidases, releasing free I-677 and FMDP inside cells . The result of antienzymic degradation test in vitro of I-677-FMDP showed that compound I-677-FMDP was resistant to the hydrolysis by carboxypeptidase A. Int J STD AIDS, 2002 Jun, 13(6), 373 - 7 Salivary IgA and serum IgA and IgG antibodies to Candida albicans in HIV-infected subjects; Belazi M et al.; This study sought to determine IgA, IgG antibodies to Candida albicans in whole saliva and serum from HIV-infected patients and to compare them to a group of healthy controls . The study population consisted of 34 HIV-infected individuals free of any other systemic diseases and thirty healthy controls . IgA concentrations in saliva and IgA and IgG concentrations in serum were measured by a micro enzyme-linked immunosorbent assay . No significant differences were observed in salivary and serum IgA antibodies to C . albicans between the two study groups . Serum IgG antibodies were found to be significantly lower in the HIV-infected (P < 0.05) . No significant changes were observed in the specific activity of anti-Candida IgA and IgG antibodies in saliva and serum, in both the study groups . The undifferentiated levels of secretory-IgA antibodies to C . albicans in the patients' and the controls' saliva could be an indicator of the high immune response to opportunistic infections of the HIV-infected subjects, a fact that is verified by the lack of oral candidiasis in the patients' group . The low levels of IgG antibodies in the serum of the HIV-infected patients confirm the high immune response of them. J Hosp Infect, 2002 Apr, 50(4), 304 - 8 Risk factors for nosocomial candidaemia: a case-control study in children; Paganini H et al.; Candida spp . are increasingly important nosocomial pathogens in critically ill children . Data on risk factors of acquisition in children with candidaemia are limited . To determine the incidence, prognosis and risk factors for acquisition of nosocomial candidaemia in children, a prospective case-control study was performed between July 1998 and January 2000 . Candidaemia was defined as the presence of at least one positive blood culture containing Candida spp . For each case, two controls were selected and matched for time and site of hospital admission . Logistic regression was used to obtain estimates of risk after simultaneous control for other variables . Candidaemia was diagnosed in 24 children (1.09/1000 admissions) . The species most frequently isolated were Candida albicans (50%) and Candida parapsilosis (17%) . Mean age was 58.2+/-58 months between cases and 41.8+/-52.2 months in controls (P not significant) . The strongest risk factors for acquisition found in the univariate analysis were underlying disease, previous antibiotic treatment, the number of antibiotics administered, presence of a long-duration catheter and total parenteral nutrition (P<0.05) . Sex, the site of hospital admission, artificial ventilatory assistance, transitory venous catheters, arterial and peripherally venous accesses, bladder catheter and transfusions were not statistically significant . The risk factors identified by multiple logistic regression analysis were: permanent catheter (OR 31.5; 3.19-310) and total parenteral nutrition (OR 10.5; 2.76-40.0) . Nine (37%) children with nosocomial candidaemia and seven (15%) controls died (P=0.05) . These findings should facilitate development of rational approaches to preventing infection and assist clinicians in identifying those patients in whom this life-threatening complication is likely to occur . Shi Yan Sheng Wu Xue Bao, 1998 Jun, 31(2), 117 - 27 {The transcriptional regulation of the TCA1 elements and their effects on morphogenesis of Candida albicans}; Wang Q et al.; We have cloned two copies of retrotransposon-like elements Tca1-1 and Tca1-2 (Transposon Candida albicans) from Candida albicans strain SC5314 . Comparing the restriction maps of the two genomic fragments, similar patterns were observed . The two composite elements were transcribed into an approximately unit length 6 kb RNA . The expression of this transcript was greatly increased when cells were grown at 25 degrees C versus 37 degrees C . Deletion of the partial or complete intervening region of the Tca1-1 element causes the morphogenetic variation of the cells . The morphogenetic variation is in cooperation with the expression of the URA3 gene. Biopolymers, 2002, 67(4-5), 362 - 6 Influence of benign cellular changes in diagnosis of cervical cancer using IR microspectroscopy; Romeo MJ et al.; IR spectroscopy and principal components analysis (PCA) of endocervical cells and smears diagnosed with benign cellular changes were investigated to determine the influence of these potential confounding variables in the diagnosis of cervical cancer . Spectral differences in all cell and diagnostic types investigated were found in the phosphodiester and carbohydrate regions . However, the spectral differences in other bands were not distinct enough to allow differentiation between groups . The PCA was successfully used to obtain a separation of normal ectocervical smears from normal endocervical cells and smears diagnosed with inflammation, Candida albicans, and bacterial vaginosis . A separation with a slight overlap of abnormal ectocervical smears from normal endocervical cells, inflammation, and bacterial vaginosis was obtained with PCA . Candida was not separated from abnormal ectocervical smears with any success . Infect Immun, 2002 Jun, 70(6), 3284 - 6 Role of interleukin-18 in host defense against disseminated Candida albicans infection; Stuyt RJ et al.; In mice injected intravenously with Candida albicans, administration of anti-interleukin-18 (IL-18) antibodies increased the yeast load in the kidneys . There was no effect on the organ load with Candida when gamma interferon (IFN-gamma)-deficient mice were treated with anti-IL-18 antibodies, suggesting that the protective effect of IL-18 is mediated through endogenous IFN-gamma. Infect Immun, 2002 Jun, 70(6), 3281 - 3 Reevaluation of the role of HWP1 in systemic candidiasis by use of Candida albicans strains with selectable marker URA3 targeted to the ENO1 locus; Sundstrom P et al.; Previous evaluation of HWP1 in systemic candidiasis in CBA/J mice was done with Candida albicans strains with differing genetic locations of URA3 as a result of Ura-blaster mutagenesis . In this study, the presence of HWP1 and the location of URA3 contributed to the severity of murine systemic candidiasis in BALB/c mice. J Agric Food Chem, 2002 May 22, 50(11), 3147 - 9 Essential oil constituents and in vitro antimicrobial activity of Decalepis hamiltonii roots against foodborne pathogens; Thangadurai D et al.; Hydrodistillation of Decalepis hamiltonii roots yielded an essential oil (0.33% v/w) that contained 2-hydroxy-4-methoxybenzaldehyde (37.45%), 2-hydroxybenzaldehyde (31.01%), 4-O-methylresorcylaldehyde (9.12%), benzyl alcohol (3.16%), and alpha-atlantone (2.06%) as major constituents, with aromatic aldehydes constituting the main fraction of this root's essential oil . The oil was tested for its antimicrobial activity against foodborne pathogens responsible for food spoilage and human pathologies using standard antimicrobial assays . It exhibited strong antimicrobial activity against Bacillus cereus, Bacillus megaterium, Candida albicans, Escherichia coli, Micrococcus luteus, Micrococcus roseus, and Staphylococcus aureus at a concentration range of 1:0 with inhibitory activities of 27, 23, 16, 19, 22, 19, and 23 mm, respectively, which are comparable to those of the standards . The roots of D . hamiltonii, therefore, may be considered as an inexpensive source of an essential oil rich in antimicrobial compounds against foodborne pathogens. J Antimicrob Chemother, 2002 May, 49(5), 731 - 40 A sub-inhibitory concentration of amphotericin B enhances candidastatic activity of interferon-gamma- and interleukin-13-treated murine peritoneal macrophages; Coste A et al.; We studied the effects of interferon-gamma (IFN-gamma), a Th1 cytokine, and interleukin-13 (IL-13) or interleukin-4 (IL-4), Th2 cytokines, on the antifungal activity of resident murine peritoneal macrophages against Candida albicans 'in vitro' . IFN-gamma, IL-13 and IL-4 treatment enhanced the candidastatic functions of the macrophages . Reactive oxygen intermediates (ROIs) seem to be directly involved in the increase of anti-Candida activity in macrophages treated with Th1 or Th2 cytokines . Study of unopsonized C . albicans phagocytosis showed that IFN-gamma reduces the uptake process whereas the Th2 cytokines increase it . This difference is correlated to mannose receptor expression, which is decreased by IFN-gamma but increased by the Th2 cytokines . So, the effects on phagocytosis and candidastatic activity of IFN-gamma-treated macrophages are dissociated . In contrast, the phagocytic ability of macrophages pretreated 'in vitro' with IL-4 or IL-13 played a complementary role to the ROIs, in reduction of yeast proliferation by macrophages . In consequence, the macrophages treated with IL-13 and IL-4 develop a higher fungistatic activity than macrophages activated by IFN-gamma . Amphotericin B associated with IL-13 or IFN-gamma, but not with IL-4, enhanced the yeast growth inhibition activity of macrophages . The ROIs were involved in the additive effect of IFN-gamma with amphotericin B, whereas another mechanism was implicated in the increase of candidastatic activity of macrophages treated with IL-13 in association with amphotericin B. Mycoses, 2002 Apr, 45(3-4), 109 - 10 Renewed increase in Candida albicans among yeast isolates from the Göttingen university hospital; Ruchel R et al.; We report on the recurrence of Candida albicans among yeast isolates from our university hospital . After a decline in occurrence which coincided with the onset of the use of fluconazole, the fraction of C . albicans recovered and at present has reached the pre-fluconazole level . No permanent rise of C . glabrata or C . krusei has been observed. Mycoses, 2002 Apr, 45(3-4), 101 - 4 Dermatophytes and other fungi associated with skin mycoses in Tripoli, Libya; Ellabib MS et al.; This study sought to determine the prevalence of skin infections and their causative agents in the Libyan population . Samples were collected from 2224 patients attending the Dermatology Clinics of the Tripoli Medical Centre (TMC) between August 1997 and December 1999 and were submitted to a mycology laboratory for analysis . Diagnosis was confirmed by microscopic examination in 1180 cases (53.1%) and the causative agent was isolated and cultured in 1160 cases (52.2%) . Dermatophytes, Malassezia furfur and Candida albicans were the most common etiological agents isolated . Tinea corporis accounted for 45.9% of cases (85% of cases occurred in children below 15 years of age) . The frequency of the other clinical types in descending order was pityriasis versicolor 27.8% (322 cases), candidiosis 13.4% (156 cases), tinea pedis 8.1% (94 cases), tinea manuum 2.6% (30 cases) and tinea barbae 2.2% (26 cases) . Trichophyton violaceum was the most common etiological agent, responsible for 44% (300 cases) of dermatophyte infections . Malassezia furfur was ranked the second most frequent causative agent being found in 27.8% of cases, followed by Trichophyton rubrum 13.8% (160 cases) and Candida albicans 10% (116 cases) . Other species isolated included Microsporum canis 8.1% (94 cases), Epidermophyton floccosum 6.6% (76 cases) and Trichophyton mentagrophytes 3.1% (36 cases). Mycoses, 2002 Apr, 45(3-4), 88 - 90 Fungistatic activity of some perfumes against otomycotic pathogens; Jain SK et al.; The sporostatic effect of five otomycotic pathogens, i.e . Aspergillus niger, A . flavus, Absidia corymbifera, Penicillium nigricans and Candida albicans to nine different perfumes was determined on the basis of their spore germination . These organisms were isolated from patients suffering from fungal infection of the external auditory canal . Volatile vapours emanating from musk, phulwari, jasmine, nagchampa and bela caused approximately 100% inhibition in spore germination of all the test fungi . Volatiles emanating from chandan, khas and hina showed no inhibition for the test pathogens, displaying their resistant character to these perfumes. Mycoses, 2002 Apr, 45(3-4), 79 - 83 The role of tumour necrosis factor-alpha (TNF-alpha) and platelet-activating factor (PAF) interaction on murine candidosis; Kalkanci A et al.; Tumour necrosis factor-alpha (TNF-alpha) is related to some other factors in addition to being the essential cytokine of the sepsis which results from Candida infections . In our study, we investigated serum TNF-alpha levels, measured by enzyme-linked immunosorbent assay (ELISA), and platelet-activating factor (PAF)-like activity, measured by high-pressure liquid chromatography (HPLC) of the mice infected with Candida species . The PAF antagonist, ginkgolide BN 52021 was used to evaluate the possible interaction between TNF-alpha and PAF . The average TNF-alpha levels were found to be 396, 489, 699 and 803 pg ml(-1) on the 4th, 5th, 6th and 19th days of Candida albicans infection, respectively (P<0.05) . There was no statistically significant difference between the serum TNF-alpha levels of the groups infected with other Candida species, such as C . kefyr, C . krusei and C . tropicalis (P>0.05) . Serum TNF-alpha levels were found to be more significantly different in mice with C . albicans infection that were injected with PAF antagonists on the 6th day (23 pg ml(-1)) . It was therefore thought that PAF antagonists have an inhibitory effect on TNF-alpha production . No significant difference was found between PAF levels in the three groups: healthy control mice, C . albicans-infected mice and C . albicans-infected mice given PAF antagonists (466 milli-absorbance unit (mAU), 475 mAU and 329 mAU, respectively) . It was noticed that the positive interaction between PAF and TNF-alpha was not important after the first 4 days of the infection had passed. Mycopathologia, 2002, 153(3), 125 - 8 Cell-associated collagenolytic activity by Candida albicans; Nishimura M et al.; Cell associated collagenolytic activity of Candida albicans was quantified by measuring the degradation of synthetic peptide 2-furanacryloyl-Leu-Gly-Pro-Ala (FALGPA), which is a specific substrate for collagenase, by the freeze-thaw procedure method . This collagenolytic activity was enhanced by cells cultured in the presence of bovine serum albumin (BSA) in culture medium . However, this activity was inhibited in the presence of ethylenediaminetetraacetic acid disodium salt (EDTA-2Na), but not by the serine proteinase inhibitor p-amidinophenyl methanesulfonyl fluoride (APMSF), nor the aspartyl proteinase inhibitor pepstatin A . These results suggested the presence of a metalloenzyme on pericellular C . albicans. Mol Microbiol, 2002 May, 44(3), 841 - 53 Complete glycosylphosphatidylinositol anchors are required in Candida albicans for full morphogenesis, virulence and resistance to macrophages; Richard M et al.; Glycosylphosphatidylinositol (GPI)-anchored proteins are involved in cell wall integrity and cell-cell interactions . We disrupted the Candida albicans homologue of the Saccharomyces cerevisiae GPI7/LAS21 gene, which encodes a GPI anchor-modifying activity . In the mutant and on solid media, the yeast-to-hyphae transition was blocked, whereas chlamydospore formation was enhanced . However, the morphogenetic switch was normal in liquid medium . Abnormal budding patterns, cytokinesis and cell shape were observed in both liquid and solid media . The cell wall structure was also modified in the mutants, as shown by hypersensitivity to Calcofluor white . In vitro and in vivo assays revealed that the mutant interacted with its host in a modified way, resulting in reduced virulence in mice and reduced survival in the gastrointestinal environment of mice . The mitogen-activated protein (MAP) kinase pathway of macrophages was downregulated by the wild-type cells but not by the DeltaCagpi7 null strains . In agreement with this abnormal behaviour, mutant cells were more sensitive to the lytic action of macrophages . Our results indicate that a functional GPI anchor is required for full hyphal formation in C . albicans, and that perturbation of the GPI biosynthesis results in hypersensitivity to host defences. Mol Microbiol, 2002 May, 44(3), 621 - 31 A basic helix-loop-helix protein with similarity to the fungal morphological regulators, Phd1p, Efg1p and StuA, controls conidiation but not dimorphic growth in Penicillium marneffei; Borneman AR et al.; Members of the APSES protein group are basic helix-loop-helix (bHLH) proteins that regulate processes such as mating, asexual sporulation and dimorphic growth in fungi . Penicillium marneffei is a human pathogen and is the only member of its genus to display a dimorphic growth transition . At 25 degrees C, P . marneffei grows with a filamentous morphology and produces asexual spores from multicellular con-idiophores . At 37 degrees C, the filamentous morphology is replaced by yeast cells that reproduce by fission . We have cloned and characterized an APSES protein-encoding gene from P . marneffei that has a high degree of similarity to Aspergillus nidulans stuA . Deletion of stuA in P . marneffei showed that it is required for metula and phialide formation during conidiation but is not required for dimorphic growth . This suggests that APSES proteins may control processes that require budding (formation of the metulae and phialides, pseudohyphal growth in Saccharomyces cerevisiae and dimorphic growth in Candida albicans) but not those that require fission (dimorphic growth in P . marneffei) . The A . nidulans DeltastuA mutant has defects in both conidiation and mating . The P . marneffei stuA gene was capable of complementing the conidiation defect but could only inefficiently complement the sexual defects of the A . nidulans mutant . This suggests that the P . marneffei gene, which comes from an asexual species, has diverged significantly from the A . nidulans gene with respect to sexual but not asexual development. J Infect Dis, 2002 May 15, 185(10), 1483 - 9 Epub 2002 Apr 30. The role of toll-like receptor (TLR) 2 and TLR4 in the host defense against disseminated candidiasis; Netea MG et al.; Toll-like receptors (TLRs) represent the main class of pattern-recognition receptors involved in sensing pathogenic microorganisms . The aim of the present study was to assess the role of TLR4 in the defense against Candida albicans infection . The outgrowth of C . albicans was 10-fold higher in TLR4-defective C3H/HeJ mice, compared with that in control C3H/HeN mice (P<.05) . Production of tumor necrosis factor (TNF) and interleukin (IL)-1alpha and IL-1beta by mouse macrophages in response to C . albicans stimulation was not affected by TLR4, and the candidacidal capacities of the neutrophils and macrophages of C3H/HeJ mice were normal . In contrast, production of the CXC chemokines KC and macrophage inhibitory protein-2 was 40%-60% lower by the macrophages of C3H/HeJ mice (P<.05), which resulted in a 40% decrease in neutrophil recruitment to the site of infection . Candida-induced TNF and IL-1beta production by human peripheral blood mononuclear cells was significantly inhibited by blocking anti-TLR2 antibodies in vitro . In conclusion, TLR4-defective C3H/HeJ mice are more susceptible to C . albicans infection, and this is associated with impaired chemokine expression and neutrophil recruitment. Can J Physiol Pharmacol, 2002 Mar, 80(3), 205 - 9 A fluorescent compound for glucose uptake measurements in isolated rat cardiomyocytes; Ball SW et al.; A focus of current diabetes research is the development of insulinomimetic compounds for oral treatment of diabetes and its associated cardiac complications . Screening compounds for their potential insulinomimetic effects usually involves the use of radioactive isotopes . The focus of this study was to investigate a nonradioactive fluorescent compound for its use in screening insulinomimetic compounds . The indicator 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) has been used by some workers to measure glucose uptake in Escherichia coli and Candida albicans . We propose that 2-NBDG will also be a suitable indicator for mammalian cell lines, in particular rat cardiomyocytes . We found that the indicator could give a reliable reproducible standard curve following appropriate dilution and is taken up by isolated cardiomyocytes . The insulinomimetic compounds vanadyl sulfate and sodium molybdate showed rates of glucose uptake similar to that of insulin . Furthermore, the rate of uptake measured for insulin using this technique (0.04 +/- 0.003 nmol x min(-1) x 10(6) cells(-1) is comparable with previous literature using 2-deoxyglucose uptake measurements on isolated myocytes (0.040 nmol x min(-1) x 10(6) cells(-1), demonstrating the validity of this fluorescent compound for glucose uptake studies. Crit Care Med, 2002 Mar, 30(3), 677 - 83 Adherence of yeast and filamentous forms of Candida albicans to cultured enterocytes; Wiesner SM et al.; OBJECTIVE: Systemic candidiasis is a major cause of complicating infections in intensive care units . Morbidity and mortality are high, even in those who receive appropriate antifungal therapy . Because the intestinal tract is considered a major portal of entry for systemic candidiasis, experiments were designed to clarify the ability of yeast and filamentous forms, as well as the INT1 gene product, to influence adherence of Candida albicans to the intestinal epithelium . DESIGN: Controlled . SETTING: University teaching hospital research laboratory . SUBJECTS: Mature Caco-2 and HT-29 cultured enterocytes . INTERVENTIONS: C . albicans INT1 mutant strains, defective in filament production, were used to observe the ultrastructural surface interactions of C . albicans with cultured intestinal epithelial cells, namely Caco-2 and HT-29 cells . These mutant strains also were used to quantify the effect of the INT1 gene product on C . albicans adherence (yeast and filamentous forms) to cultured enterocytes . Ultrastructural surface interactions of C . albicans with cultured enterocytes were observed with high resolution scanning electron microscopy . C . albicans adherence to cultured enterocytes was quantified by using a colorimetric enzyme-linked immunosorbent assay . MEASUREMENTS AND MAIN RESULTS: Both yeast and filamentous forms of C . albicans appeared tightly adherent to the apical surface of cultured enterocytes, and INT1 appeared to have little, if any, effect on these ultrastructural surface interactions . The distal ends of C . albicans filaments appeared to mediate adherence to enterocyte apical microvilli, and thigmotropism (contact guidance) appeared to play a role in C . albicans adherence . The absence of functional INT1 was associated with decreased adherence of C . albicans yeast forms to cultured enterocytes . CONCLUSIONS: Although functional INT1 appeared to facilitate adherence of C . albicans yeast forms to cultured enterocytes, the role of INT1 in adherence of filamentous forms was unclear, and both yeast and filamentous forms could adhere to, and perhaps invade, the apical surface of cultured enterocytes. J Med Microbiol, 2002 May, 51(5), 433 - 42 Combined detection of mannanaemia and antimannan antibodies as a strategy for the diagnosis of systemic infection caused by pathogenic Candida species; Sendid B et al.; A novel strategy for the diagnosis of systemic candidosis was evaluated, based on the combination of two enzyme immunoassays that detect a candida oligomannoside repetitive epitope expressed in large amounts by Candida albicans (Platelia Candida Ag), and antibodies against C . albicans mannan, the major cell-wall immunogen in which this epitope is present (Platelia Candida Ab) . Sera were selected retrospectively from intensive care and haematology patients with clinically suspected systemic candidosis, and from whom Candida spp . had been isolated from normally sterile sites . Of the 21 patients infected with C . albicans, 13 had positive antigenaemia and 14 had a positive antibody response, including eight patients who were antigenaemia negative . The sensitivity of the combined tests was 100% . In patients infected with C . glabrata (n = 12) or C . tropicalis (n = 10), the sensitivity was 83% and 80%, respectively . For the remaining patients, infected with C . parapsilosis (n = 10), C . krusei (n = 8) or C . kefyr (n = 2), the sensitivity of the combined tests was 40%, 50% and 50%, respectively . At least one of the serological tests was positive before yeast growth occurred in 60% of patients for whom a serum sample was available before blood culture sampling . An increase in serological test positivity to >80% was observed for sera obtained around the date of positive culture, irrespective of the Candida species isolated . These results suggest that regular serological monitoring for both mannanaemia and anti-mannan antibodies in at-risk patients may contribute to the early diagnosis of candidosis. J Pediatr Hematol Oncol, 2002 Mar-Apr, 24(3), 237 - 9 Candida dubliniensis fungemia and vascular access infection; Cimolai N et al.; Candida dubliniensis is a newly recognized species of yeast, which may have been forrmerly identified as Candida albicans, that has been rarely isolated from invasive fungal infections among humans . The authors document a C . dubliniensis fungemia that occurred during the course of a vascular access infection in a 2-year-old who was undergoing active therapy for neuroblastoma . Presumptive C . albicans isolates from an 18-year period were reassessed, and it was found that C . dubliniensis is a rare cause of fungemia among pediatric patients (0.5% of all such isolates). Planta Med, 2002 Apr, 68(4), 363 - 5 Antifungal metabolite with a new carbon skeleton from Keissleriella sp . YS4108, a marine filamentous fungus; Liu CH et al.; In addition to four known metabolites (4-acetyl-6,8-dihydroxy-5-methylisocoumarin, 6,8-dihydroxy-3-methylisocoumarin, 6,8-dihydroxy-3,5,7-trimethylisocoumarin and 3,3'-oxy-(5-methyl)-phenol), bioassay-guided fractionation of the culture of Keissleriella sp., a marine filamentous fungus (strain number: YS 4108), afforded an antifungal metabolite with a new carbon skeleton whose structure was elucidated spectrometrically as 3,6,8-trihydroxy-3-{3,5-dimethyl-2-oxo-3(E)-heptenyl}-2,3-dihydronaphthalen-1(4H)-one . In vitro antifungal assays of all isolates revealed that the new metabolite and 3,3'-oxybis{5-methylphenol} were inhibitory to the growth of the human pathogenic fungi Candida albicans, Tricophyton rubrum and Aspergillus niger with MICs of the former being 40, 20 and 80 microg/ml, and those of the latter 10, 30 and 50 microg/ml, respectively. Microbiology, 2002 May, 148(Pt 5), 1291 - 303 The ARO4 gene of Candida albicans encodes a tyrosine-sensitive DAHP synthase: evolution, functional conservation and phenotype of Aro3p-, Aro4p-deficient mutants; Sousa S et al.; The enzyme 3-deoxy-D-arabinoheptulosonate-7-phosphate (DAHP) synthase catalyses the first step in aromatic amino acid biosynthesis in prokaryotes, plants and fungi . Cells of Saccharomyces cerevisiae contain two catalytically redundant DAHP synthases, encoded by the genes ARO3 and ARO4, whose activities are feedback-inhibited by phenylalanine and tyrosine, respectively . ARO3/4 gene transcription is controlled by GCN4 . The authors previously cloned an ARO3 gene orthologue from 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) a homozygous aro3-deletion mutant of C . albicans is phenotypically Aro(+), suggesting the existence of another isozyme(s) . They now report the identification and functional characterization of the C . albicans orthologue of S . cerevisiae Aro4p . The two Aro4p enzymes share 68% amino acid identity . Phylogenetic analysis places the fungal DAHP synthases in a cluster separate from prokaryotic orthologues and suggests that ARO3 and ARO4 arose from a single gene via a gene duplication event early in fungal evolution . C . albicans ARO4 mRNA is elevated upon amino acid starvation, consistent with the presence of three putative Gcn4p-responsive elements (GCREs) in the gene promoter sequence . C . albicans ARO4 complements an aro3 aro4 double mutation in S . cerevisiae, an effect inhibited by excess tyrosine . The authors engineered Deltaaro3/Deltaaro3 Deltaaro4/MET3p::ARO4 cells of C . albicans (with one wild-type copy of ARO4 placed under control of the repressible MET3 promoter) and found that they fail to grow in the absence of aromatic amino acids when ARO4 expression is repressed, and that this growth defect can be partially rescued by aromatic amino acids and certain aromatic amino acid pathway intermediates . It is concluded that, like S . cerevisiae, C . albicans contains two DAHP synthases required for the first step in the aromatic amino acid biosynthetic pathway. Microbiology, 2002 May, 148(Pt 5), 1281 - 90 Disruption in Candida albicans of the TPS2 gene encoding trehalose-6-phosphate phosphatase affects cell integrity and decreases infectivity; Zaragoza O et al.; The gene CaTPS2 encoding trehalose-6-phosphate (T6P) phosphatase from Candida albicans has been cloned and disrupted in this organism . The Catps2/Catps2 mutant did not accumulate trehalose but accumulated high levels of T6P . Disruption of the two copies of the CaTPS2 gene did not abolish growth even at 42 degrees C, but decreased the growth rate . In the stationary phase, the Catps2/Catps2 mutant aggregated, more than 50% of its cells became permeable to propidium iodide and a large amount of protein was found in the culture medium . Aggregation occurred only at pH values higher than 7 and was avoided by osmoprotectants; it was never observed during the exponential phase of growth . The mutant formed colonies with a smooth border on Spider medium . Mice inoculated with 1.5 x 10(6) c.f.u . of wild-type cells died after 8 days, while 80% of those inoculated with the same number of c.f.u . of the Catps2/Catps2 mutant survived for at least 1 month . Reintroduction of the wild-type CaTPS2 gene in the Catps2/Catps2 mutant abolished the phenotypes described . It is hypothesized that the accumulation of T6P interferes with the assembly of a normal cell wall. Clin Exp Immunol, 2002 May, 128(2), 365 - 71 Highly active antiretroviral therapy restores in vitro mitogen and antigen-specific T-lymphocyte responses in HIV-1 perinatally infected children despite virological failure; Peruzzi M et al.; To analyse the effect of highly active antiretroviral therapy (HAART) on T-lymphocyte functions we selected seven HIV-1 perinatally infected children (CDC immunological category 1 or 2) who had neither a fall in their plasma HIV-1 RNA levels nor a significant rise in CD4+ lymphocyte counts while receiving HAART . Clinical signs and symptoms were monitored monthly . Plasma viral load, CD4+, CD8+, CD19+ lymphocyte counts and in vitro T-lymphocyte proliferative responses to mitogens (anti-CD3, phytohaemoagglutinin, concanavalin A and pokeweed mitogen) and recall antigens (Candida albicans and tetanus toxoid) were tested at baseline and after 1, 3, 6 and 12 months of HAART . Twenty-two healthy age-matched children were studied as controls . A gain in body weight, no worsening of the disease and no recurrence of opportunistic infections were observed . At baseline, the majority of the children had low responses to mitogens, and all of them had a defective in vitro antigen-specific T-lymphocyte response (<2 standard deviations below the mean result for controls) . During HAART, a significant increase in the response to mitogens and antigens was observed in all the patients . The T-lymphocyte response was restored more consistently against antigens to which the immune system is constantly exposed (Candida albicans, baseline versus 12 months: P < 0.001) compared with a low-exposure antigen (tetanus toxoid, baseline versus 12 months: P < 0.01) . HAART restores in vitro T-lymphocyte responses even in the absence of a significant viral load decrease and despite any significant increase in CD4+ lymphocyte counts . It implies that a direct mechanism might be involved in the overall immune recovery under HAART. Rev Neurol (Paris), 2002 Apr, 158(4), 473 - 6 {Candida albicans meningoencephalomyeloradiculitis}; Tiberghien F et al.; A 25-year-old immunocompetent male heroin addict was admitted for acute confusion associated with gait disorders of three month duration . The diagnosis was meningoencephalomyeloradiculitis secondary to Candida albicans infection . Outcome was good after a 6-month regimen with antifungal drugs . Neurological complications of Candida albicans infection are rare and prognosis is generally poor . This case report illustrates diagnostic and therapeutic difficulties encountered. Eur J Immunol, 2002 May, 32(5), 1455 - 63 CD40/CD40 ligand interactions in the host defense against disseminated Candida albicans infection: the role of macrophage-derived nitric oxide; Netea MG et al.; CD40L interaction with CD40 is required for normal cellular immune responses such as T cell-mediated activation of monocytes/macrophages, proinflammatory cytokine production, and leukocyte extravasation . We investigated the role of CD40/CD40 ligand (L) interactions during disseminated candidiasis in CD40L knockout (CD40L-/-) mice . While early during infection there were no differences in the Candida albicans outgrowth in the organs of wild-type and knockout mice, the CD40L-/- mice had a significantly increased yeast load in the kidneys compared to CD40L+/+ mice late during infection . Similar effects were observed in CD40L+/+ mice in which CD40 ligation was blocked by a neutralizing anti-CD40 antibody . The peak TNF-alpha plasma concentrations were significantly lower in the CD40L-/- mice than in CD40L+/+ mice . C . albicans-stimulated production of nitric oxide (NO) by peritoneal macrophages from CD40L-/- in vitro was significantly lower than that of control mice, and this was responsible for a reduced candidacidal activity of CD40L-/- macrophages . The role of endogenous NO synthesis induced by CD40 ligation for the defense against disseminated candidiasis was further demonstrated by the absence of these effects in knockout mice deficient in inducible NO-synthase . In conclusion, absence of CD40/CD40L interactions results in increased susceptibility to disseminated infection with C . albicans through decreased NO-dependent killing of Candida by macrophages. Eur J Nucl Med Mol Imaging, 2002 May, 29(5), 674 - 9 Epub 2002 Mar 06. Technetium-99m labelled fluconazole and antimicrobial peptides for imaging of Candida albicans and Aspergillus fumigatus infections; Lupetti A et al.; The aim of this study was to investigate whether technetium-99m labelled fluconazole can distinguish fungal from bacterial infections . Fluconazole was labelled with (99m)Tc and radiochemical analysis showed less than 5% impurities . The labelling solution was injected into animals with experimental infections . For comparison, we used two peptides for infection detection, i.e . UBI 29-41 and hLF 1-11, and human IgG, all labelled with (99m)Tc . Mice were infected with Candida albicans or injected with heat-killed C . albicans or lipopolysaccharides to induce sterile inflammation . Also, mice were infected with Staphylococcus aureus or Klebsiella pneumoniae . Next, accumulation of (99m)Tc-fluconazole and (99m)Tc-labelled peptides/IgG at affected sites was determined scintigraphically . (99m)Tc-fluconazole detected C . albicans infections (T/NT ratio=3.6+/-0.47) without visualising bacterial infections (T/NT ratio=1.3+/-0.04) or sterile inflammatory processes (heat-killed C . albicans: T/NT ratio=1.3+/-0.2; lipopolysaccharide: T/NT ratio=1.4+/-0.1) . C . albicans infections were already seen within the first hour after injection of (99m)Tc-fluconazole (T/NT ratio=3.1+/-0.2) . A good correlation (R(2)=0.864; P<0.05) between T/NT ratios for this tracer and the number of viable C . albicans was found . Although (99m)Tc-UBI 29-41 and (99m)Tc-hLF 1-11 were able to distinguish C . albicans infections from sterile inflammatory processes in mice, these (99m)Tc-labelled peptides did not distinguish these fungal infections from bacterial infections . It is concluded that (99m)Tc-fluconazole distinguishes infections with C . albicans from bacterial infections and sterile inflammations. Genetics, 2002 Apr, 160(4), 1749 - 53 Invasive filamentous growth of Candida albicans is promoted by Czf1p-dependent relief of Efg1p-mediated repression; Giusani AD et al.; Filamentation of Candida albicans occurs in response to many environmental cues . During growth within matrix, Efg1p represses filamentation and Czf1p relieves this repression . We propose that Czf1p interacts with Efg1p, altering its function . The complex regulation of filamentation may reflect the versatility of C . albicans as a pathogen. Int J STD AIDS, 2002 May, 13(5), 318 - 22 Microscopy of stained smears of vaginal secretion in the diagnosis of recurrent vulvovaginal candidosis; Novikova N et al.; This study was undertaken to determine the value of stained smears from the vaginal introitus and the posterior vaginal fornix for detection of candida morphotypes in the diagnosis of recurrent vulvovaginal candidosis (RVVC) in cases with an assumed novel attack of the condition, and to compare the value of microscopy of such smears in relation to candida culture, vaginal pH-determinations and leucocyte esterase tests (LE) . One hundred and thirteen women with a history and a current assumed attack of RVVC were studied by means of culture of samples from the vaginal introitus and posterior vaginal fornix on Sabouraud and CHROMagar . Microscopy of methylene blue- and Gram-stained smears from these sites was performed . The pH of vaginal secretion and the LE activity in vaginal flushing fluid was measured . Candida morphotypes were found significantly more often in the smears from candida culture-positive than culture-negative women . There was no difference in this respect between the findings in the methylene blue- and Gram-stained smears . Differences were found in candida morphotypes among Candida albicans and non-albicans-positive cases, as blastoconidia were detected only in the latter cases . The leukocyte esterase activity score was higher in the candida-positive than in candida-negative cases . The study showed that microscopy of fixed, stained genital smears can play a role in attempts to distinguish cases infected and not infected by candida among patients consulting with an assumed novel attack of RVVC . Study of methylene blue-stained smears is recommended as they represent an easier means than Gram-stained smears to diagnose genital candida infections in assumed RVVC cases. Toxicol Appl Pharmacol, 2002 Apr 15, 180(2), 136 - 44 Dermal application of jet fuel suppresses secondary immune reactions; Ramos G et al.; Applying military jet fuel (JP-8) to the skin of mice activates systemic immune suppression . In all of our previous experiments, JP-8 was applied to immunologically naive mice . The effect of jet fuels on established immune reactions, such as immunological memory, is unknown . The focus of the experiments presented here was to test the hypothesis that jet fuel exposure {both JP-8 and commercial jet fuel (Jet-A)} suppresses established immune reactions . Mice were immunized with the opportunistic fungal pathogen Candida albicans and, at different times after immunization (10 to 30 days), various doses of undiluted JP-8 or Jet-A were applied to their skin . Both the elicitation of delayed-type hypersensitivity (DTH) (mice challenged 10 days after immunization) and immunological memory (mice challenged 30 days after immunization) were significantly suppressed in a dose-dependent manner . Dermal exposure to either multiple small doses (50 microl over 4 days) or a single large dose (approximately 200-300 microl) of JP-8 and/or Jet-A suppressed DTH to C . albicans . The mechanism by which dermal application of JP-8 and Jet-A suppresses immunological memory involves the release of immune biologic response modifiers . Blocking the production of prostaglandin E(2) by a selective cyclooxygenase-2 inhibitor (SC 236) significantly reversed jet fuel-induced suppression of immunologic memory . These findings indicate, for the first time, that dermal exposure to commercial jet fuel (Jet-A) suppresses the immune response . In addition, the data reported here expand on previous findings by suggesting that jet fuel exposure may depress the protective effect of prior vaccination . (c)2002 Elsevier Science (USA). Yeast, 2002 May, 19(7), 611 - 8 Construction of FLAG tagging vectors for Candida albicans; Umeyama T et al.; We have constructed three new vectors for Candida albicans (pFLAG-Act1, pFLAG-Mal2, and pFLAG-Met3) . The proteins can be expressed as C-terminal FLAG-tagged proteins under the control of different promoters (ACT1, MAL2, and MET3) . To confirm the protein expression, we used the Renilla reniformis luciferase and the drug efflux pump Cdr1p of Candida albicans as reporters . The luciferase protein expressed by the MET3 promoter was found to have the strongest activity of the three promoters when cultured in a methionine-depleted synthetic medium . Cdr1p was expressed as a C-terminal FLAG-tagged protein using either these vectors or PCR-mediated integration . The fluconazole resistance was increased by the Cdr1p expression in a CDR1 homozygous disruptant . The expressed proteins were detected by Western blotting using the anti-FLAG antibody . We also constructed a Cdr1p-FLAG expressing strain, in which we directly tagged Cdr1p with FLAG on the genome loci, using a PCR-based integrative marker cassette that was amplified using the pFLAG vector . We then confirmed the protein expression by Western blotting . Thus, these new vectors are useful as C . albicans genetic tools . Mol Microbiol, 2002 Apr, 44(1), 61 - 72 Candida albicans Als1p: an adhesin that is a downstream effector of the EFG1 filamentation pathway; Fu Y et al.; Filamentation and adherence to host cells are critical virulence factors of Candida albicans . Multiple filamentation regulatory pathways have been discovered in C . albicans using Saccharomyces cerevisiae as a model . In S . cerevisiae, these pathways converge on Flo11p, which functions as a downstream effector of f