Candida albicans

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 filamentation and also mediates cell-cell adherence (flocculation) . In C . albicans, such effector(s) have not yet been identified . Here, we demonstrate that the cell surface protein Als1p is an effector of filamentation in C . albicans . We show that Als1p expression is controlled by the transcription factor Efg1p, which is known to be a key regulator of filamentation in C . albicans . Further, disruption of ALS1 inhibited filamentation, and autonomous expression of Als1p restored filamentation in an efg1 homozygous null mutant . Thus, Als1p functions as a downstream effector of the EFG1 filamentation pathway . In addition, we found that Als1p mediates both flocculation and adherence of C . albicans to endothelial cells in vitro . As a cell surface glycoprotein that mediates filamentation and adherence, Als1p has both structural and functional similarity to S . cerevisiae Flo11p . Consistent with our in vitro results, Als1p was required for both normal filamentation and virulence in the mouse model of haematogenously disseminated candidiasis.

Curr Opin Infect Dis, 2002 Apr, 15(2), 143 - 7
New perspectives on the immunology of chronic mucocutaneous candidiasis; Lilic D; Chronic mucocutaneous candidiasis is a primary immune deficiency presenting as an inability to clear fungal infections and consequently as persisting and recurring infections of the skin and mucous membranes with yeasts, mostly Candida albicans . Chronic mucocutaneous candidiasis is a heterogeneous clinical syndrome which usually presents in childhood and can have an autosomal recessive, dominant or sporadic mode of inheritance . Most chronic mucocutaneous candidiasis patients also develop accompanying endocrine and inflammatory disorders that suggest an underlying deregulation of the immune system . It has long been recognized that protection from mucocutaneous candidiasis relies on cell-mediated immunity and studies on animal models have highlighted the essential role of type 1 cytokines in protection against Candida spp . Recent data in patients with chronic mucocutaneous candidiasis have documented altered patterns of cytokine production in response to Candida spp . with decreased production of some but not all type 1 cytokines and increased levels of interleukin-10 . The defect underlying altered cytokine production remains unknown but studies are in progress addressing the putative role of dendritic cells and pattern recognition receptors in directing cytokine responses . These novel insights into immune mechanisms responsible for protection against Candida spp . are opening new possibilities of immunomodulation and vaccination that could prove beneficial in the management of chronic mucocutaneous candidiasis.

Curr Opin Infect Dis, 2001 Dec, 14(6), 673 - 7
The role of yeasts other than Candida albicans in oropharyngeal candidiasis; Redding SW; Candida albicans has been recognized as the predominant infecting organism in oropharyngeal candidiasis . Yeasts other than C . albicans are commonly recognized as colonizing the oral cavity but have not been thought to be a significant cause of disease . This review will describe the emergence of yeasts other than C . albicans as causative pathogens in oropharyngeal candidiasis both as co-infecting organisms with C . albicans and as sole pathogens themselves . Diagnosis and treatment of these emerging infections will also be discussed.

Phytomedicine, 1999 Nov, 6(5), 357 - 62
Complement-mediated antiinflammatory effect of bisbenzylisoquinoline alkaloid fangchinoline; Hristova M et al.; Complement-mediated mode of action of bisbenzylisoquinoline alkaloid fangchinoline was investigated in vivo and in vitro . The application of fangchinoline intraperitoneally (i.p.) to complement normal mice, strain ICR, inhibited the complement activity in serum and peritoneal exudate . The substance activated serum complement of C5-deficient DBA/2 mice . Fangchinoline was able to provoke local inflammatory reaction in both strains after subcutaneous (s.c.) injection . The alkaloid suppressed paw swelling induced by live Candida albicans in ICR and DBA/2 mice . Its effect depended on the dose and time of injection prior to inflammatory reaction . The in vitro experiments proved the interference of fangchinoline action with post-C5 reactions . The substance augmented C5-convertase formation and functional activity . These results are in correspondence with our previous investigations, proving the complement-mediated action of fangchinoline . The antiinflammatory effect could be a consequence of the caused complement exhaustion.

Mol Biol Evol, 2002 May, 19(5), 664 - 77
Ylli, a non-LTR retrotransposon L1 family in the dimorphic yeast Yarrowia lipolytica; Casaregola S et al.; During the course of a random sequencing project of the genome of the dimorphic yeast Yarrowia lipolytica, we have identified sequences that were repeated in the genome and that matched the reverse transcriptase (RT) sequence of non-long terminal repeat (non-LTR) retrotransposons . Extension of sequencing on each side of this zone of homology allowed the definition of an element over 6 kb long . The conceptual translation of this sequence revealed two open reading frames (ORFs) that displayed several characteristics of non-LTR retrotransposons: a Cys-rich motif in the ORF1, an N-terminal endonuclease, a central RT, and a C-terminal zinc finger domain in the ORF2 . We called this element Ylli (for Y . lipolytica LINE) . A total of 19 distinct repeats carrying the 3' untranslated region (UTR) and all ending with a poly-A tail were detected . Most of them were very short, 17 being 134 bp long or less . The number of copies of Ylli was estimated to be around 100 if these short repeats are 5' truncations . No 5' UTR was clearly identified, indicating that entire and therefore active elements might be very rare in the Y . lipolytica strain tested . Ylli does not seem to have any insertion specificity . Phylogenetic analysis of the RT domain unambiguously placed Ylli within the L1 clade . It forms a monophyletic group with the Zorro non-LTR retrotransposons discovered in another dimorphic yeast Candida albicans . BLAST comparisons showed that ORF2 of Ylli is closely related to that of the slime mold Dictyostelium discoideum L1 family, TRE.

Antimicrob Agents Chemother, 2002 May, 46(5), 1447 - 54
A chemically modified tetracycline (CMT-3) is a new antifungal agent; Liu Y et al.; Several chemically modified tetracycline analogs (CMTs), which were chemically modified to eliminate their antibacterial efficacy, were unexpectedly found to have antifungal properties . Of 10 CMTs screened in vitro, all exhibited antifungal activities, although their efficacies varied . Among these compounds, CMT-315, -3, and -308 were found to be the most potent as antifungal agents . The MICs of CMT-3 against 47 strains of fungi in vitro were determined by using amphotericin B (AMB) and doxycycline as positive and negative controls, respectively . The MICs of CMT-3 were generally found to be between 0.25 and 8.00 microg/ml, a range that approximates the blood levels of this drug when administrated orally to humans . Of all the yeast species tested to date, Candida albicans showed the greatest sensitivity to CMT-3 . The filamentous species most susceptible to CMT-3 were found to be Epidermophyton floccosum, Microsporum gypseum, Pseudallescheria boydii, a Penicillium sp., Scedosporium apiospermum, a Tricothecium sp., and Trichophyton rubrum . Growth inhibition of C . albicans by CMT-3, determined by a turbidity assay, indicated a 50% inhibitory concentration of 1 microg/ml . Thirty-nine strains, including 20 yeasts and 19 molds, were used to measure viability (the ability to grow after treatment with a drug) inhibition by CMT-3 and AMB . CMT-3 exhibited fungicidal activity against most of these fungi, especially the filamentous fungi . Eighty-four percent (16 of 19) of the filamentous fungi tested showed more than 90% inhibition of viability by CMT-3 . In contrast, AMB showed fungicidal activity against all yeasts tested . However, most of the filamentous fungi (16 of 19) showed less than 50% inhibition of viability by AMB, indicating that AMB is fungistatic against most of these filamentous fungi . To begin to identify the sites in fungal cells affected by CMT-3, C . albicans and a Penicillium sp . were incubated with the compound at 35 degrees C, and then the fluorescence of CMT-3 was observed by confocal laser scanning electron microscopy . CMT-3 appeared to have widespread intracellular distribution throughout C . albicans and the Penicillium sp . The mechanisms of the antifungal activity of CMT-3 are now being explored.

Antimicrob Agents Chemother, 2002 May, 46(5), 1240 - 5
Efficacies of fluconazole, caspofungin, and amphotericin B in Candida glabrata-infected p47phox-/- knockout mice; Ju JY et al.; Candida glabrata is the second leading cause of adult candidemia, resulting in high mortality . Amphotericin B is considered the treatment of choice, while the efficacy of fluconazole is controversial and caspofungin efficacy is unknown . To ascertain drug efficacy in vivo, the utility of a murine model of C . glabrata infection was investigated . C . glabrata was found to cause progressive, lethal infection when injected intravenously into C57BL/6 mice with reduced oxidative microbicidal capacity due to knockout of the p47(phox) gene . Spleen and kidney organ CFU counts were determined in groups of mice 2 days after the mice completed 6 days of daily intraperitoneal drug treatment, which began on the day of infection . Daily injections of fluconazole at 80 mg/kg did not reduce spleen or kidney CFU counts after infection with C . glabrata strains having in vitro fluconazole MICs of 2, 32, or 256 microg/ml compared to saline-treated controls . However, this fluconazole regimen reduced spleen CFU counts in mice infected with Candida albicans, an infection that is known to be responsive to fluconazole . Caspofungin at 5 mg/kg and amphotericin B at 5 mg/kg were both effective in reducing fungal burden in spleens and kidneys of C . glabrata-infected mice . Ten mice treated for 6 days with caspofungin at 1 mg/kg survived for 15 days, though all 10 saline-injected mice died or were so ill that they had to be sacrificed by 96 h postinfection . This murine model provided evidence of the efficacy of amphotericin B and caspofungin but not of fluconazole against C . glabrata infection.

Bioorg Med Chem Lett, 2002 Mar 25, 12(6), 943 - 6
Oxime derivatives of sordaricin as potent antifungal agents; Serrano-Wu MH et al.; Oxime derivatives of the sordarin aglycone have been identified as potent antifungal agents . The in vitro spectrum of activity includes coverage against Candida albicans and Candida glabrata with MICs as low as 0.06 microg/mL . The antifungal activity was established to be exquisitely sensitive to the spatial orientation of the lipophilic side chains.

Bioorg Med Chem Lett, 2002 Mar 25, 12(6), 895 - 8
Evaluation of morphogenic regulatory activity of farnesoic acid and its derivatives against Candida albicans dimorphism; Kim S et al.; A series of farnesoic acid derivatives was prepared and their morphogenic regulatory activities were evaluated . Their inhibitory activities against yeast cell growth and yeast-to-hypha transition examined in Candida albicans cells are dependent upon the chain length as well as the substitution patterns on the isoprenoid template . The preliminary structure-activity relationship of these compounds is described to elucidate the essential structural requirements.

Ethiop Med J, 1998 Oct, 36(4), 235 - 43
Characterization of Candida albicans isolates from the oral cavity of HIV-positive patients; Woldeamanuel Y et al.; Characterization of Candida albicans isolates from the oral cavity of Ethiopian Human Immunodeficiency Virus (HIV)-positive patients and HIV-negative individuals was undertaken . After identification of isolates using conventional methods and determination of carbohydrate assimilation profiles, serotyping was performed by slide agglutination with the Iatron IF6 serotyping system . Sensitivity of the isolates to 5-Fluorocytosine (5-FC) was assessed using the broth macrodilution method . There was no difference in phenotype, by any of the three methods used, between the HIV-positive and HIV-negative individuals . Serotype A was found to be dominant in strains isolated from both HIV-positive (34/40 = 85%) and HIV-negative (17/20 = 85%) study subjects . These data suggest that the C . albicans strains causing oral candidiasis in patients with HIV infection are identical to the commensal ones found in healthy individuals.

Infect Immun, 2002 May, 70(5), 2725 - 9
Intravaginal and intranasal immunizations are equally effective in inducing vaginal antibodies and conferring protection against vaginal candidiasis; De Bernardis F et al.; Oophorectomized, estrogen-treated rats were immunized by the intravaginal or intranasal route with a mannoprotein extract (MP) or secreted aspartyl proteinases (Sap) of Candida albicans, with or without cholera toxin as a mucosal adjuvant . Both routes of immunization were equally effective in (i) inducing anti-MP and anti-Sap vaginal antibodies and (ii) conferring a high degree of protection against the vaginal infection by the fungus . These data suggest that appropriate fungal antigens and adjuvant can be used to protect against candidal vaginitis, by either route.

Acta Paediatr, 2002, 91(2), 198 - 202
Risk factors for invasive fungal infection in neonates; El-Masry FA et al.; Invasive fungal infection is an uncommon, but increasing cause of morbidity and mortality in neonates . There are few controlled studies defining risk factors for the development of fungal infection in a contemporary neonatal population . This retrospective case-control study was undertaken to investigate antenatal, demographic and postnatal variables that may be potentially important in the development of fungal infection . Two gestation-matched controls were identified for each index case . Information about perinatal and demographic variables, as well as important neonatal outcomes, was obtained from case notes . Microbiological data collected included the presence of fungal colonization, and organisms responsible for invasive fungal infection . Over a 5-y period, 24 infants with invasive fungal infection and 48 controls were identified . Candida albicans was the organism identified in 75% of cases of fungal septicaemia, and in all cases complicated by fungal meningitis . Preceding fungal colonization, pulmonary haemorrhage and intrauterine growth restriction were factors significantly and independently associated with invasive fungal infection . Fifty-four percent of infants with invasive fungal infection died, and 82% of survivors developed chronic lung disease . CONCLUSION: Some new and potentially important risk factors for the development of invasive fungal infection in a contemporary population of infants admitted to a neonatal intensive care were identified.

J Perinatol, 2002 Apr-May, 22(3), 204 - 8
Dexamethasone therapy and Candida sepsis in neonates less than 1250 grams; Pera A et al.; OBJECTIVE: To determine whether dexamethasone use increases the risk for Candida sepsis (CS) in very low birth weight premature infants (<1250 g) . DESIGN: Retrospective chart review of all infants with a birth weight <1250 g, admitted to the neonatal intensive care unit of the MetroHealth Medical Center, Cleveland, Ohio between January 1, 1996 and December 31, 1999 . Infant groups with (n=65) and without (n=229) CS were compared . RESULTS: Two hundred and ninety four infants with a birth weight <1250 g were identified . CS was diagnosed at a median age of 18 days, and 6 of 65 (10%) infants died directly from Candida-related complications . Candida albicans (n=30, 60%) and Candida parapsilosis (n=14, 25%) were the predominant isolates . Use of dexamethasone in infants at risk for chronic lung disease before 14 days of age (p=0.001), duration of antibiotics (p=0.001), and total duration of parenteral nutrition and intralipid (p=0.0001) were all significantly greater in infants who developed CS . Regression analysis showed that duration of antibiotics before the diagnosis of Candida infection (r(2)=0.69, p=0.0002) and duration of dexamethasone (r(2)=0.93, p=0.0002) correlated with Candida infection . Early dexamethasone use was also related to the age at diagnosis of Candida infection (r(2)=0.51, p=0.01) . CONCLUSIONS: Dexamethasone therapy and prolonged duration of antibiotics are associated with Candida infection in premature infants.

J Immunol, 2002 Apr 15, 168(8), 4055 - 62
C/EBP beta gene inactivation causes both impaired and enhanced gene expression and inverse regulation of IL-12 p40 and p35 mRNAs in macrophages; Gorgoni B et al.; The transcription factor C/EBPbeta is believed to play a fundamental role in regulating activated macrophage functions . However, the molecular mechanisms and the target genes involved have been, so far, poorly characterized, partly due to the difficulty of reproducibly obtaining homogeneous and abundant primary macrophage populations . In this study, we describe the generation and characterization of immortalized macrophage-like cell lines from C/EBPbeta-deficient and wild-type mice . Using these cells, we were able to identify a number of genes involved in activated macrophage functions whose induction was affected in the C/EBPbeta(-/-) cells . IFN-gamma/LPS-dependent induction of IL-6, IL-1beta, TNF-alpha, inducible NO synthase, and plasminogen activator inhibitor-1 mRNAs was variably impaired, while IL-12 p40, RANTES and macrophage inflammatory protein-1beta mRNAs were up-regulated in the absence of C/EBPbeta . The differential mRNA expression correlated with differential transcription levels of the corresponding genes, and was in most cases confirmed in primary macrophage populations . Moreover, in sharp contrast to the enhanced induction of IL-12 p40 mRNA, C/EBPbeta(-/-) primary macrophages derived from both the bone marrow and the peritoneal cavity displayed totally defective expression of IL-12 p35 mRNA . Therefore, the IL-12 p35 gene represents a novel obligatory target for C/EBPbeta in macrophages and this may explain the defective production of bioactive IL-12 and the impaired Th1 responses of C/EBPbeta-deficient mice to Candida albicans infection observed in previous work.

J Biol Chem, 2002 Jul 12, 277(28), 25512 - 8 Epub 2002 Apr 05.
Identification and characterization of a sphingolipid delta 4-desaturase family; Ternes P et al.; Sphingolipids desaturated at the Delta4-position are important signaling molecules in many eukaryotic organisms, including mammals . In a bioinformatics approach, we now identified a new family of protein sequences from animals, plants, and fungi and characterized these sequences biochemically by expression in Saccharomyces cerevisiae . This resulted in the identification of the enzyme sphingolipid Delta4-desaturase (dihydroceramide desaturase) from Homo sapiens, Mus musculus, Drosophila melanogaster, and Candida albicans, in addition to a bifunctional sphingolipid Delta4-desaturase/C-4-hydroxylase from M . musculus . Among the sequences investigated are the Homo sapiens membrane lipid desaturase, the M . musculus degenerative spermatocyte, and the Drosophila melanogaster degenerative spermatocyte proteins . During spermatogenesis, but not oogenesis of des mutant flies, both cell cycle and spermatid differentiation are specifically blocked at the entry into the first meiotic division, leading to male sterility . This mutant phenotype can be restored to wild-type by complementation with a functional copy of the des gene (Endo, K., Akiyama, T., Kobayashi S., and Okada, M . (1996) Mol . Gen . Genet . 253, 157-165) . These results suggest that Delta4-desaturated sphingolipids provide an early signal necessary to trigger the entry into both meiotic and spermatid differentiation pathways during Drosophila spermatogenesis.

Bioorg Med Chem, 2002 Jun, 10(6), 1681 - 6
Anti-Candida albicans properties of novel benzoxazine analogues; Fringuelli R et al.; We previously reported that azole 1,4-benzothiazine derivatives have appreciable anti-Candida activity . In this study, we synthesized 1,4-benzoxazine analogues and examined their possible antifungal activity to further analyze the structure-activity relationships . Results of in vitro and in vivo experiments showed that 1,4-benzoxazine analogues show appreciable antifungal activity . In particular, they have significant capability to cure mice systemically infected with a lethal challenge of Candida albicans, as indicated by increased survival time paralleling reduction of colony forming units . Moreover, 1,4-benzoxazine derivatives also showed immunomodulating activity, as indicated by a significant increase of interleukin-12 and interferon-gamma production by splenocytes and reinforcement of a T helper type 1 protective immune response to C . albicans . In conclusion, the results demonstrate that replacement of sulfur by oxygen may improve immune response against C . albicans infection.

Curr Genet, 2002 Feb, 40(5), 326 - 38 Epub 2002 Feb 06.
Nep1p (Emg1p), a novel protein conserved in eukaryotes and archaea, is involved in ribosome biogenesis; Eschrich D et al.; The essential Saccharomyces cerevisiae gene YLR186w { NEP1 (nucleolar essential protein 1), also recently named EMG1 (essential for mitotic growth 1)} is highly conserved in eukaryotes and archaea . In S . cerevisiae, Nep1p is localized in the nucleus with a rod-shaped morphology, which first suggested a spindle/microtubule association for ScNEP1 (S . cerevisiae NEP1) . However, in Candida albicans and in HeLa cells, Nep1p is a protein of the nucleoli . CaNEP1 (C . albicans NEP1) and HsNEP1 (Homo sapiens NEP1) heterologously complement the essential phenotype in a S . cerevisiae nep1 deletion mutant . However, the ScNEP1 spindle/microtubule phenotype is not found with HsNEP1 and CaNEP1, which shows that the spindle/microtubule association is specific for ScNep1p and that it is not the primary essential function of Nep1p . A temperature-sensitive ScNEP1 (ts1) allele was isolated and revealed a strongly increased sensitivity to paromomycin, a translational inhibitor which binds to RNA, indicating that ribosome biogenesis within the nucleolus is probably affected . This was confirmed by polysome profile and ribosomal subunit analysis, showing a ribosomal subunit imbalance with a decrease in 40S subunits, due to reduced amounts of native 18S rRNA . Furthermore, ScSAM2, encoding S-adenosylmethionine synthetase, was isolated as a multicopy-suppressor of the ScNEP1 (ts1) mutant allele and medium supplementation with S-adenosylmethionine restored its growth . For the first time, these results identified a novel protein with an essential function in ribosomal biogenesis which directly or indirectly interferes with a methylation reaction during the early steps of pre-rRNA processing necessary for the generation of 40S ribosomal subunits.

Phytother Res, 2002 Mar, 16 Suppl 1, S93 - 6
Bioactivity studies on Cassia alata Linn . leaf extracts; Villasenor IM et al.; The hexane, chloroform and ethyl acetate extracts of the leaves of Cussia alata Linn . were tested for their antimutagenic, antifungal, analgesic, antiinflammatory and hypoglycaemic activities . At a dosage of 5 mg/20 g mouse, the hexane extract was analgesic as it reduced the number of squirms induced by acetic acid by 59.5% . Both the hexane and EtOAc extracts exhibited antiinflammatory activity at a dosage of 5 mg/20 g mouse with a 65.5% and 68.2% decrease in carrageenan-induced inflammation, respectively . The chloroform extract was antimutagenic, at a dosage of 2 mg/20 g mouse, with a 65.8% inhibition in the mutagenicity of tetracycline . It was also the most active against Trichophyton mentagrophytes, at a concentration of 50 mg/mL but it had no activity against Candida albicans . The hexane and EtOAc extracts showed some activity against both organisms, with the EtOAc extract being more active against C . albicans . The EtOAc extract was hypoglycaemic . At a dosage of 5 mg/20 g mouse, it decreased the blood sugar level of mice by 58.3% . Pharmacological studies showed that all extracts caused an immediate decrease in motor activity, enophthalmus, hyperemia, micturition and diarrhoea . At a dosage of 150 mg/20 g mouse, the EtOAc extract caused paralysis, screen grip loss and enophthalmus accompanied by drooping and closure of the eyelids .

Microbiology, 2002 Apr, 148(Pt 4), 1073 - 9
Complications in cell-surface labelling by biotinylation of Candida albicans due to avidin conjugate binding to cell-wall proteins; Masuoka J et al.; Initial contact between the opportunistic fungal pathogen Candida albicans and host tissue occurs at the cell surface . Biotin derivatives have been used to label the cell-surface proteins of yeasts, with labelled proteins subsequently detected by avidin-reporter conjugates . Previous work has indicated that avidin can bind to C . albicans proteins in the absence of biotin, suggesting a possible host-cell-recognition mechanism by fungal cell-surface proteins . To investigate this mechanism, Western blots of proteins extracted from biotinylated and mock-treated cells were probed with avidin or modified-avidin reagents . Each avidin reagent bound to cell-wall proteins extracted from non-biotinylated cells . Binding did not appear to be due to the lectin-like activity of the cell-wall proteins of C . albicans or to the presence of biotin in the sample itself . Binding was inhibited by added biotin, by the chaotrope KSCN and by NaCl in a concentration-dependent manner, although inhibition varied among the avidin conjugates tested . Thus, the non-specific binding of avidin to the cell-wall proteins of C . albicans appears to involve hydrophobic and electrostatic interactions, depending on the particular avidin species . These observations demonstrate potential pitfalls in the use of avidin-biotin complexes to identify cell-surface molecules and could provide insights into protein-protein interactions at the C . albicans cell wall.

Microbiology, 2002 Apr, 148(Pt 4), 1061 - 72
Homozygosity at the Candida albicans MTL locus associated with azole resistance; Rustad TR et al.; Antifungal drug resistance in the pathogenic fungus Candida albicans is a serious threat to the growing population of immunocompromised patients . This study describes a significant correlation between loss of heterozygosity at the C . albicans mating-type-like (MTL) locus and resistance to azole antifungals . A pool of 96 clinical isolates consisting of 50 azole-resistant or susceptible dose-dependent isolates and 46 azole-susceptible isolates was screened by PCR for the presence of MTLa1 and MTLalpha1 . These genes were used as markers for the MTLa and MTLalpha loci . Both loci were present in 84 of the isolates . Six isolates failed to amplify MTLa1 and six failed to amplify MTLalpha1 . Further PCR analysis demonstrated that loss of the MTLa1 and MTLalpha1 genes corresponded to loss of all of the loci-specific genes, resulting in homozygosity at the MTL locus . Southern analysis and single nucleotide polymorphism (SNP) analysis were used to determine that this loss of heterogeneity was due to replacement of one of the MTL loci with a duplicate of the other locus resulting in two homozygous copies of the MTL locus . Of the 12 homozygous isolates, one isolate was sensitive to azole drugs . Statistical analysis of the data demonstrates a strong correlation between homozygosity at the MTL locus and azole resistance (P<0 small middle dot003) . In a set of serial isolates, an increase in azole resistance correlated with the loss of heterozygosity at the MTL locus, lending further strength to the correlation . Gene disruptions of the MTL loci were found to have no effect on azole susceptibility.

Microbiology, 2002 Apr, 148(Pt 4), 1049 - 60
The Candida albicans Sup35p protein (CaSup35p): function, prion-like behaviour and an associated polyglutamine length polymorphism; Resende C et al.; The Sup35p protein of Saccharomyces cerevisiae is an essential translation factor whose prion-like properties give rise to the non-Mendelian genetic element {PSI(+)} . In this study the SUP35 gene from the related yeast species Candida albicans has been characterized . The CaSUP35 gene encodes a protein (CaSup35p) of 729 aa which shows 65% amino acid identity to the S . cerevisiae Sup35p protein (ScSup35p), with the C-terminal region showing greater identity (79%) than the N-terminal region . The full-length CaSup35p can functionally replace ScSup35p in S . cerevisiae although complementation is only complete when CaSup35p is overexpressed . Complementation only requires expression of the CaSup35p C domain . In S . cerevisiae the full-length CaSup35p is unable to establish a prion-like aggregated state even in the presence of endogenous ScSup35p prion 'seeds', thus confirming the existence of a species barrier in fungal prion propagation . Subcellular localization studies in C . albicans show that although CaSup35p is normally ribosome-associated, when not ribosome-associated, it does not form pelletable high-molecular-mass aggregates characteristic of the ScSup35p in {PSI(+)} strains . Unlike the ScSup35p, the CaSup35p N domain contains a number of polyglutamine repeats although it does contain seven copies of the peptide GGYQQ that is repeated in the ScSup35p N domain . Analysis of the CaSUP35 gene from 14 different strains of C . albicans identified four naturally occurring polymorphisms associated with changes in the length of the largest of the polyglutamine repeats . These findings have important implications for the evolution of fungal prion genes.

J Infect Dis, 2002 Apr 15, 185(8), 1103 - 14 Epub 2002 Mar 21.
Mucosal candidiasis in transgenic mice expressing human immunodeficiency virus type 1; de Repentigny L et al.; The availability of CD4C/HIV(MutA) transgenic (Tg) mice expressing human immunodeficiency virus type 1 in immune cells and developing an AIDS-like disease has provided the opportunity to devise a model of mucosal candidiasis that closely mimics the clinical and pathologic features of candidal infection in human AIDS . After intraoral infection with Candida albicans, oral burdens were strikingly elevated in the Tg mice, compared with non-Tg littermates (P<.05), during primary infection, a 6-10-week carrier state, and a marked terminal outgrowth preceding death . The chronic carrier state was absent in the non-Tg mice because of clearing of C . albicans . Candida hyphae penetrated the epithelium of the oral cavity, esophagus, and cardial-atrium fold of the stomach, accompanied by a mononuclear cell infiltrate . Immunohistochemical analysis suggested that decreased frequencies of major histocompatibility complex class II-expressing cells, combined with reduced CD4+ cells, may underlie the susceptibility to mucosal candidiasis in these Tg mice.

J Infect Dis, 2002 Apr 15, 185(8), 1094 - 102 Epub 2002 Mar 21.
Evidence for differential expression of candida albicans virulence genes during oral infection in intact and human immunodeficiency virus type 1-transgenic mice; Ripeau JS et al.; To comprehensively assess the in vivo expression of Candida albicans hydrolytic enzyme genes during oropharyngeal candidiasis (OPC), a controlled sequential analysis of the temporal expression of individual members of the SAP (secretory aspartyl proteinase) gene family and PLB1 (phospholipase B) in a murine model of OPC was conducted . Acute infections in intact C3H and DBA/2 mice were terminated by clearance of C . albicans within 7 days after oral inoculation, but transgenic (Tg) mice expressing human immunodeficiency virus type 1 were persistently colonized until a final outgrowth before death . In contrast to the sustained expression of other SAP genes and PLB1, SAP7 and SAP8 were conspicuously distinguished by their transient expression in both intact and Tg mice . SAP5 and SAP9 were most strongly expressed throughout the course of infection in the Tg mice . These findings indicate that expression of individual members of the C . albicans SAP gene family is differentially regulated during experimental OPC.

Diagn Microbiol Infect Dis, 2002 Mar, 42(3), 181 - 5
Comparison of Albicans ID2 agar plate with the germ tube for presumptive identification of Candida albicans; Cardenes CD et al.; Albicans ID2 (bioMerieux, France) is a commercially available chromogenic medium that allows rapid and specific macroscopic identification of Candida albicans and facilitates the differentiation of species in mixed cultures . We compared it with the standard method for the identification of yeast species, the germ tube test (GT) . This study involved 423 clinical isolates, including 163 C . albicans and 260 non-albicans yeasts . Sensitivity of Albicans ID2 agar plates regarding the identification of C . albicans were 98.2% after 48 h of incubation and specificity of 96.6% . This method using rapid enzymatic method shows the same similar sensitivity than the GT test The false negative rate (1.8%) for the GT test is consistent with that previously reported . None tests discriminated between C . albicans and C . dubliniensis isolates.

Oral Microbiol Immunol, 2002 Apr, 17(2), 129 - 31
Differences in Candida albicans adhesion to intact and denatured type I collagen in vitro; Makihira S et al.; An inhibition assay of Candida albicans adhesion to gelatin-immobilized membranes was compared with that to intact type I collagen-immobilized membranes using an arginine-glycine-aspartic acid (RGD) containing peptide . As compared with a protein-free membrane, gelatin and collagen significantly enhanced the adherence of C . albicans . The adhesion of the yeast to gelatin was significantly inhibited by the RGD peptides, but not by arginine-glycine-glutamic acid (RGE) peptides . In contrast, attachment to collagen was not inhibited by RGD peptides . These results suggest that the RGD sequence of gelatin and the integrin-like proteins of yeasts may be involved in adherence.

Oral Microbiol Immunol, 2002 Apr, 17(2), 89 - 94
Identification of Candida dubliniensis among oral yeast isolates from an Italian population of human immunodeficiency virus-infected (HIV+) subjects; Giammanco GM et al.; Candida dubliniensis, an emerging oral pathogen, phenotypically resembles Candida albicans so closely that it is easily misidentified as such . The aim of the present study was to evaluate the usefulness of two phenotypic methods, growth at 45 degrees C and 2,3,5-triphenyltetrazolium chloride (TTC) reduction, for confirming presumptive identification of C . dubliniensis and C . albicans by colony color on CHROMagar Candida (CAC) medium . A combination of these methods was used to establish the prevalence of oral C . dubliniensis in an Italian population of 45 human immunodeficiency virus (HIV)-infected subjects . Twenty-two samples (48.9%) were positive for yeasts on CAC medium producing a total of 37 fungal isolates . The colony color and 45 degrees C growth ability test correctly identified all C . dubliniensis and C . albicans isolates (5/37, 13.5%, and 16/37, 43.2%, respectively), while assessment of TTC reduction misidentified one C . albicans isolate . The isolation rate of C . dubliniensis was 11.1% (5/45 patients) . All of the C . dubliniensis isolates were highly susceptible to fluconazole (MIC = 0.5 microg/ml) . The combination of CAC medium screening with growth at 45 degrees C and TTC reduction tests may represent a simple, reliable and inexpensive identification protocol for C . dubliniensis.

Oral Microbiol Immunol, 2002 Apr, 17(2), 79 - 84
Oral yeast carriage in patients with advanced cancer; Davies AN et al.; The aim of this study was to investigate oral yeast carriage amongst patients with advanced cancer . Oral rinse samples were obtained from 120 subjects . Yeasts were isolated using Sabouraud's dextrose agar and CHROMagar Candida, and were identified using a combination of the API 20 C AUX yeast identification system, species-specific PCR and 26S rDNA gene sequencing . Oral yeast carriage was present in 66% of subjects . The frequency of isolation of individual species was: Candida albicans, 46%; Candida glabrata, 18%; Candida dubliniensis, 5%; others, < 5% . The increasing isolation of non-Candida albicans species is clinically important, since these species are often more resistant to antifungal drugs . Oral yeast carriage was associated with denture wearing (P = 0.006), and low stimulated whole salivary flow rate (P = 0.009) . Identification of these risk factors offers new strategies for the prevention of oral candidosis in this group of patients.

Mol Microbiol, 2002 Feb, 43(3), 571 - 84
Functional analysis of a vacuolar ABC transporter in wild-type Candida albicans reveals its involvement in virulence; Theiss S et al.; ATP-driven transport proteins belonging to the ATP-binding cassette (ABC) superfamily perform important functions in cell metabolism and detoxification . Compounds can be actively transported across membranes, including the plasma membrane or organellar membranes . The vacuole is an important organelle in fungal cells required for compartmentalization of metabolites as well as toxic substances . Sequestration into the vacuole is often energy-dependent . We present the first isolation and molecular analysis of a vacuolar ABC transporter gene in the opportunistic fungal pathogen Candida albicans . The protein encoded by the MLT1 gene is highly similar to Multiple Drug Resistance-associated Protein (MRP)-like transporters of yeast and higher organisms that form the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR)/MRP subfamily of ABC transporters, a class of proteins so far not characterized in C . albicans . MLT1 expression is extensively growth phase-regulated, and gene transcripts are inducible by metabolic poisons . Gene replacement mutants generated in wild-type C . albicans with the dominant selection marker MPAR showed a profound reduction in virulence in a mouse peritonitis model that was reversed by complementation with an intact MLT1 gene . Hence, this report provides primary evidence for the involvement of vacuolar ABC transporters in fungal virulence.

Scand J Infect Dis, 2002, 34(2), 145 - 7
Candida albicans endocarditis diagnosed by PCR-based molecular assay in a critically ill pediatric patient; Posteraro B et al.; A young Down's syndrome patient developed tricuspid valve endocarditis several years after undergoing surgical closure of a congenital ventricular septal defect . Fungal etiology was established by PCR amplification of the Candida albicans ERG11 gene . Although antifungal therapy was administered, surgical replacement of the infected valve was required to eliminate the infection.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2002 Mar, 93(3), 281 - 6
Candidal carriage in the oral cavity of human immunodeficiency virus-infected subjects; Campisi G et al.; OBJECTIVES . We sought to assess asymptomatic oral carriage of Candida species and relative density in human immunodeficiency virus-infected (HIV+) subjects, as well as to determine whether yeast carriage is associated with CD4+ cell count, HIV-1 RNA quantity, gender, route of HIV transmission, antiretroviral therapy, or smoking . STUDY DESIGN . A cross-sectional analysis of oral rinses from HIV+ and healthy subjects was undertaken . Oral candidal carriage and relative species were investigated in 42 HIV+ and 41 healthy individuals, all of whom were seen at the Palermo University Department of Oral Sciences, Italy . Data were managed and analyzed by means of the computer software package StatView 5.0.1 . RESULTS . Carriage rate was 61.9% and 29.3% (P =.003) among HIV+ and healthy subjects, respectively . Similarly, density carriage in the HIV+ group was found to be significantly higher than in the control group (P = .0002) . Among HIV+ subjects, Candida carriage was significantly associated with smoking, whereas density was not . Oral candidal carriage and relative density were not significantly associated with the other 4 parameters evaluated . Candida albicans was the most frequently recovered species . CONCLUSIONS . Asymptomatic candidal carriage and relative density were found to be significantly higher in the oral cavity of HIV+ subjects-but not associated with CD4+ counts or HIV-1 RNA quantities.

J Clin Microbiol, 2002 Apr, 40(4), 1508 - 10
Prospective evaluation of the new chromogenic medium Candida ID, in comparison with Candiselect, for isolation of molds and isolation and presumptive identification of yeast species; Letscher-Bru V et al.; We conducted a prospective evaluation of Candida ID chromogenic medium (bioMerieux, Marcy l'Etoile, France) with 786 clinical specimens in comparison with Candiselect medium (Bio-Rad, Marnes la Coquette, France) . Candida ID chromogenic medium identified 97.7% of Candida albicans strains; enabled presumptive identification of C . tropicalis, C . lusitaniae, C . guillermondii, and C . kefyr and better detection of yeast combinations (11.4% more often); and was more sensitive for the isolation of filamentous fungi (17.7% more often) . However, Candida ID chromogenic medium appeared to be less selective vis-a-vis bacteria, with bacterial colonies sometimes pigmented blue.

J Clin Microbiol, 2002 Apr, 40(4), 1298 - 302
Epidemiology of candidemia: 3-year results from the emerging infections and the epidemiology of Iowa organisms study; Diekema DJ et al.; Bloodstream infections due to Candida species cause significant morbidity and mortality . Surveillance for candidemia is necessary to detect trends in species distribution and antifungal resistance . We performed prospective surveillance for candidemia at 16 hospitals in the State of Iowa from 1 July 1998 through 30 June 2001 . Using U.S . Census Bureau and Iowa Hospital Association data to estimate a population denominator, we calculated the annual incidence of candidemia in Iowa to be 6.0 per 100,000 of population . Candida albicans was the most common species detected, but 43% of candidemias were due to species other than C . albicans . Overall, only 3% of Candida species were resistant to fluconazole . However, Candida glabrata was the most commonly isolated species other than C . albicans and demonstrated some resistance to azoles (fluconazole MIC at which 90% of the isolates tested are inhibited, 32 microg/ml; 10% resistant, 10% susceptible dose dependent) . C . glabrata was more commonly isolated from older patients (P = 0.02) and caused over 25% of candidemias among persons 65 years of age or older . The investigational triazoles posaconazole, ravuconazole, and voriconazole had excellent in vitro activity overall against Candida species . C . albicans is the most important cause of candidemia and remains highly susceptible to available antifungal agents . However, C . glabrata has emerged as an important and potentially antifungal resistant cause of candidemia, particularly among the elderly.

J Clin Microbiol, 2002 Apr, 40(4), 1290 - 7
Usefulness of multilocus sequence typing for characterization of clinical isolates of Candida albicans; Bougnoux ME et al.; Molecular characterization of Candida albicans isolates is essential for understanding the epidemiology of nosocomial infections caused by this yeast . Here, we investigated the potential value of multilocus sequence typing (MLST) for characterizing epidemiologically related or unrelated C . albicans strains of various clinical origins . Accordingly, we sequenced the internal regions (loci) of six selected housekeeping genes of 40 C . albicans clinical isolates and 2 reference strains . In all, 68 polymorphic nucleotide sites were identified, of which 65 were found to be heterozygous in at least one isolate . Ten to 24 different genotypes were observed at the different loci, resulting, when combined, in 39 unique genotype combinations or diploid sequence types (DSTs) . When MLST was applied to 26 epidemiologically unrelated isolates and the 2 reference strains, it allowed the identification of 27 independent DSTs, thus demonstrating a discriminatory power of 99.7 . Using multidimensional scaling together with the minimum spanning tree method to analyze interstrain relationships, we identified six groups of genetically related isolates on the basis of bootstrap values of greater than 900 . Application of MLST to 14 epidemiologically related isolates showed that those recovered from patients in the same hospital ward during the same 3 months had specific DSTs, although 73% of these isolates were genetically very close . This suggests that MLST can trace minute variations in the sequences of related isolates . Overall, MLST proved to be a highly discriminatory and stable method for unambiguous characterization of C . albicans.

J Mass Spectrom, 2002 Mar, 37(3), 265 - 9
Mechanism of azole antifungal activity as determined by liquid chromatographic/mass spectrometric monitoring of ergosterol biosynthesis; Heimark L et al.; A liquid chromatography/mass spectrometry (LC/MS) method for separation and characterization of ergosterol biosynthetic precursors was developed to study the effect of Posaconazole on sterol biosynthesis in fungi . Ergosterol biosynthetic precursors were characterized from their electron ionization mass spectra acquired by a normal-phase chromatography, particle beam LC/MS method . Fragment ions resulting from cleavage across the D-ring and an abundant M - 15 fragment ion were diagnostic for methyl substitution at C-4 and C-14 . Comparison of the sterol profile in control and treated Candida albicans incubations showed depletion of ergosterol and accumulation of C-4 and C-14 methyl-substituted sterols following treatment with Posaconazole . These C-4 and C-14 methyl sterols are known to be incapable of sustaining cell growth . The results demonstrate that Posaconazole exerts its antifungal activity by inhibition of ergosterol biosynthesis . Furthermore, Posaconazole appears to disrupt ergosterol biosynthesis by inhibition of lanosterol 14alpha-demethylase .

Yeast, 2002 Mar 30, 19(5), 423 - 8
Functional characterization of the Candida albicans homologue of secretion-associated and Ras-related (Sar1) protein; Jiang L et al.; Secretion-associated and Ras-related protein (Sar1p) plays an essential role during the protein transport from the endoplasmic reticulum to the Golgi apparatus . The cDNA sequence of the Sar1 gene has been identified and characterized from the human yeast pathogen, Candida albicans . This cDNA encodes a protein of 190 amino acids, which shares a 78% sequence identity with Saccharomyces cerevisiae Sar1p and contains the conserved GTP-binding motifs of the small GTPase superfamily . Complementation studies confirmed that this cDNA encodes the functional homologue of ScSar1p . The recombinant C . albicans Sar1p exhibits GTP-binding activity in vitro that was abolished by deletion of one of the three GTP-binding motifs .

J Infect Dis, 2002 Apr 1, 185(7), 963 - 70 Epub 2002 Mar 11.
The role of endogenous interleukin (IL)-18, IL-12, IL-1beta, and tumor necrosis factor-alpha in the production of interferon-gamma induced by Candida albicans in human whole-blood cultures; Netea MG et al.; Despite the importance of interferon (IFN)-gamma, tumor necrosis factor (TNF), and interleukin (IL)-18 for host defense against candidiasis, the pathways leading to their stimulation by Candida albicans are unclear . In a whole-blood model, IL-18 neutralization by IL-18 binding protein decreased C . albicans-induced IFN-gamma synthesis by 72% . Similarly, neutralization of IL-12 or IL-1beta by either neutralizing antibodies or IL-1 receptor antagonist also reduced (by 65%) IFN-gamma production . Neutralization of TNF by TNF binding proteins resulted in only a 36% reduction of IFN-gamma synthesis . In contrast, production of TNF and IL-8 was largely unaffected by these cytokine inhibitors . Thus, C . albicans stimulates IFN-gamma production in an IL-18-, IL-12-, and IL-1beta-dependent manner, whereas production of TNF and IL-8 is independent of these cytokines . Blocking the biologic activities of IL-18, IL-12, and IL-1beta in patients (e.g., for treatment of autoimmune diseases) may result in increased susceptibility to C . albicans infection.

Mol Microbiol, 2002 Mar, 43(5), 1197 - 214
A common drug-responsive element mediates the upregulation of the Candida albicans ABC transporters CDR1 and CDR2, two genes involved in antifungal drug resistance; de Micheli M et al.; Upregulation of the ATP-binding cassette (ABC) transporter genes CDR1 and CDR2 (Candida drug resistance 1 and 2) is a common mechanism observed in Candida albicans clinical isolates developing resistance to the class of azole antifungals . In this work, the regulatory elements of both genes were delimited using a reporter system in an azole-susceptible strain exposed to oestradiol, which allows transient induction of these genes . We found two regulatory elements in the CDR1 promoter: one responsible for basal expression (basal expression element; BEE) and the other required for oestradiol responsiveness (drug-responsive element I; DREI) . In the CDR2 promoter, a single regulatory element responsible for oestradiol responsiveness (DREII) was detected . Both DREs shared a consensus of 21 bp with the sequence 5'-CGGA(A/T)ATCGGATATTTTTTTT-3' having no equivalent to known eukaryotic regulatory sequence . Consistent with this finding, two other C . albicans genes identified by a search for the presence of DRE in the C . albicans genome sequence database were responsive to oestradiol . Finally, the regulatory elements found in CDR1 and CDR2 were also functional in an azole-resistant strain with constitutive high expression of both transporters . These results suggest that, although CDR1 and CDR2 upregulation can be obtained by transient drug-induced and constitutive upregulation, these two processes converge to the same regulatory elements and probably mobilize the same trans-acting factors.

J Invest Dermatol, 2002 Apr, 118(4), 652 - 7
Infection of human oral epithelia with Candida species induces cytokine expression correlated to the degree of virulence; Schaller M et al.; A defined and balanced immunomodulatory response is crucial for the protection of mucosal surfaces being in contact with pathogenic microorganisms . This study examined the local host response mechanisms of epithelial cells in experimental Candida albicans, C . tropicalis, and C . glabrata infections by measuring the expression of cytokines at the mRNA and protein level . During the course of infection with active but not with heat-killed C . albicans stimulation of the gene expression levels for interleukin-1alpha, interleukin-1beta, tumor necrosis factor, Exodus-2, P-selectin ligand, granulocyte-monocyte colony-stimulating factor, and interleukin-8 was observed by standard and quantitative reverse transcription-polymerase chain reaction . This cytokine pattern may favor a chemotactic and a T helper 1 response . Initial moderate or weak upregulation of these cytokine genes by reverse transcription-polymerase chain reaction was also observed in epithelial infection with the less virulent species C . tropicalis and C . glabrata . Heat-killed C . albicans failed to induce an epithelial immune response . At the protein level, expression of interleukin-8 protein was strongly enhanced during the course of C . albicans infection, whereas lower levels were seen with C . tropicalis and C . glabrata . The different expression patterns of cytokines were associated with differences in virulence of the Candida strains . This study's data, therefore, show a correlation between the virulence potential of pathogenic fungi, possibly mediated by specific virulence factors (such as proteinases), and the secretion of epithelial cytokines and chemokines, which may initiate in vivo a protective T helper 1 immunologic response and contribute to the recruitment of activated leukocytes and lymphocytes to the site of mucosal infection.

Planta Med, 2002 Mar, 68(3), 281 - 3
Antispasmodic and antimicrobial diterpenic acids from Viguiera hypargyrea roots; Zamilpa A et al.; Two spasmolytic diterpene acids, ent-beyer-15-en-19-oic acid and ent-kaur-16-en-19-oic acid, have been isolated from the roots of Viguiera hypargyrea by bioassay-guided fractionation . Beyerenoic acid was also active against Staphylococcus aureus, Enterococcus feacalis and Candida albicans.

Biochemistry, 2002 Apr 2, 41(13), 4425 - 36
Interactions of the antimicrobial peptides temporins with model biomembranes . Comparison of temporins B and L; Zhao H et al.; Temporins are short (10-13 amino acids) and linear antimicrobial peptides first isolated from the skin of the European red frog, Rana temporaria, and are effective against Gram-positive bacteria and Candida albicans . To get insight into their mechanism(s) of action, we compared the effects on model membranes exerted by two members of this family, viz., temporin B (LLPIVGNLLKSLL-NH(2)) and temporin L (FVQWFSKFLGRIL-NH(2)) . More specifically, we measured their insertion into lipid monolayers as well as their effects on the structural dynamics of liposomal bilayers as revealed by diphenylhexatriene (DPH)- and pyrene-labeled phospholipids . We also observed the impact of these peptides on the topology of giant vesicles . Both temporins readily penetrate into lipid monolayers, their intercalation being enhanced in the presence of the common bacterial negatively charged phospholipid phosphatidylglycerol . Instead, the eukaryotic lipid cholesterol did to some extent counteract their penetration into the lipid films . Both temporin B and temporin L caused an enrichment of phospholipids in the bilayers, and in the presence of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), these peptides increased acyl chain order . Temporin B had practically no effect on giant liposomes composed of 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), whereas rapid vesiculation was observed when POPG was present . In contrast, temporin L induced vesiculation of both SOPC and SOPC/POPG giant vesicles while the presence of cholesterol in SOPC giant vesicles attenuated this effect.

Mycopathologia, 2002, 153(1), 5 - 10
Cloning of Aspergillus fumigatus histidine kinase gene fragment and its expression during invasive infection; Du C et al.; It was demonstrated recently that three histidine kinases genes in Candida albicans contributed to virulence, indicating the importance of signaling pathways regulated by histidine kinases . In the present study, using a set of degenerate primers, RT-PCR was performed with cDNA of A . fumigatus as a template . PCR products were cloned and sequenced . After Blast analysis, it was found that one fragment (named as AFHK1), 305 bp, was highly homologous to the two-component histidine kinase tesA gene of Aspergillus nidulans . But AFHKI was not completely identical to the FOS-1 gene of A . fumigatus . The same A . fumigatus strain was used to inoculate the mice for a murine model of invasive pulmonary aspergillosis (IPA) . After 5-days post-inoculation, the lungs of infected animals were removed and incubated for 2 h at 37 degrees C in digestion buffer containing collagenase and trypsin . The pulmonary cells were removed by passing the suspension through a sieve . The non-filterable hyphae were treated with deoxygenated sodium cholate . Total RNA of A . fumigatus isolated from the infected tissues or cultured in vitro was extracted . With AFHKI as a probe . a Northern blot was performed . A 3.0 kb (approximate) transcript of mRNA was detected corresponding to the putative histidine kinase gene . It was demonstrated that that gene was expressed at markedly higher levels in vivo than in vitro . The results suggest that this gene may contribute to the survival and virulence of A . fumigatus.

Mycopathologia, 2002, 153(1), 25 - 8
The non-steroidal anti-inflammatory drug niflumic acid inhibits Candida albicans growth; Baker A et al.; The non-steroidal anti-inflammatory drug niflumic acid was found to inhibit growth of the yeast form of Candida albicans . Niflumic acid inhibited respiratory oxygen uptake and it is hypothesised that this was achieved by cytosolic acidification and block of glycolysis . Inhibitory concentrations are compatible with current practice of topical application.

Trends Microbiol, 2002 Apr, 10(4), 173 - 7
Anti-fungal therapy at the HAART of viral therapy; Munro CA et al.; HIV-positive patients receiving combination therapy (highly active anti-retroviral treatment, HAART) suffer significantly fewer oral infections with the opportunistic fungal pathogen Candida albicans than non-HAART-treated patients . One component of HAART is an inhibitor of the HIV proteinase, the enzyme required for correct processing of retroviral precursor proteins . It would appear that HIV proteinase inhibitors also have a direct effect on one of the key virulence factors of C . albicans, the secreted aspartic proteinases (Saps) . This suggests that the reduction in C . albicans infections in HIV-positive patients might not be solely the result of improved immunological status but could also be caused by the HAART treatment directly inhibiting Candida proteinases.

Am J Med, 2002 Apr 1, 112(5), 380 - 5
The epidemiology of Candida glabrata and Candida albicans fungemia in immunocompromised patients with cancer; Bodey GP et al.; PURPOSE: Candida glabrata is an increasing cause of candidemia, especially at cancer and bone marrow transplant centers where fluconazole is used for antifungal prophylaxis . This yeast is less susceptible to fluconazole in vitro than is Candida albicans . We compared the characteristics of patients who had C . glabrata and C . albicans candidemia at a large cancer center . SUBJECTS AND METHODS: We searched the microbiological laboratory reports and identified 116 cases of C . glabrata candidemia between 1993 and 1999 . The 116 cases of C . albicans candidemia that occurred most closely in time (before or after each case of C . glabrata candidemia) served as the control group . Data were collected from patients' medical records . RESULTS: When compared with patients who had C . albicans infection, patients with C . glabrata candidemia more often had an underlying hematologic malignancy (68 {59%} vs . 26 {22%}, P = 0.0001), had an Acute Physiology and Chronic Health Evaluation (APACHE) II score > or =16 (55 {48%} vs . 28 {25%}, P = 0.0002), and received fluconazole prophylaxis (57 {49%} vs . 8 {7%}, P = 0.0001) . Patients with C . albicans candidemia more often had concomitant infections (101 {87%} vs . 78 {67%}, P = 0.0003) and septic thrombophlebitis (11 {10%} vs . 2 {2%}, P = 0.01) . Among patients treated with antifungal therapy, those with C . albicans candidemia had a significantly greater overall response to therapy (83/104 {80%} vs . 60/97 {62%}, P = 0.005) and to primary therapy (74/104 {71%} vs . 45/97 {46%}, P = 0.0003) . Amphotericin B preparations were not more effective than fluconazole (19/45 {42%} vs . 20/38 {53%}, P = 0.5) in patients with C . glabrata candidemia . Fluconazole was less effective against C . glabrata than against C . albicans (20/38 {53%} vs . 57/74 {77%}, P = 0.008) . CONCLUSION: C . glabrata has emerged as an important cause of candidemia, especially among neutropenic patients who receive fluconazole prophylaxis.

Clin Otolaryngol, 2002 Feb, 27(1), 22 - 6
Clinical and pathological characteristics of oral lichen planus in hepatitis C-positive and -negative patients; Romero MA et al.; The reported prevalence rate of anti-hepatitis C virus (HCV) antibodies in patients with oral lichen planus shows wide geographical variation and ranges from 0 to 65% . Certain characteristic clinical features have been attributed to oral lichen planus associated to HCV infection . The purpose of this investigation has been to assess hypothetical clinical differences, as well as differences in the intensity of the subepithelial inflammatory infiltrate between oral lichen planus-HCV +ve patients and oral lichen planus-HCV -ve patients . A total of sixty-two patients entered the study . Their mean age was 63.5 +/- 14.49 years, and 48.4% of them were men and 51.6% women . Patients were classified according to their serum HCV positivity . Age, sex, clinical presentation (reticular or atrophic-erosive), extension of the lesions, location of the lesions, number of locations affected, intensity of the inflammatory infiltrate and Candida albicans colonization were recorded for each patient . Reticular lichen planus was the most frequent clinical presentation in both HCV +ve (57.1%) and HCV -ve patients (63.6%) . C . albicans colonization ranged from 42.8% in HCV +ve and 41.7% in HCV -ve patients . HCV + ve patients showed certain oral locations more frequently affected than HCV -ve ones: lip mucosa, 28.6% versus 7.3%; tongue, 57.1% versus 29.1%; and gingiva, 71.4% versus 23.6% . The number of affected intraoral locations was higher in HCV +ve patients (71.4%) than among HCV -ve ones (20.4%; chi2 = 8.34; P < 0.011) . No statistically significant differences could be established in terms of density of subepithelial inflammatory infiltrate between the groups . Our results reinforce the need for liver examination in all patients with oral lichen planus, particularly those showing lesions on the gingiva with multiple intraoral locations affected, as no pathological differences could be identified between HCV + ve and HCV -ve patients.

Farmaco, 2002 Feb, 57(2), 171 - 4
Synthesis and antimicrobial activity of some new hydrazones of 4-fluorobenzoic acid hydrazide and 3-acetyl-2,5-disubstituted-1,3,4-oxadiazolines; Rollas S et al.; A series of hydrazide hydrazones and 1,3,4-oxadiazolines of 4-fluorobenzoic acid hydrazide were prepared and evaluated as potential antimicrobial agents and were tested for their antibacterial and antifungal activities against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans . From these compounds, 4-fluorobenzoic acid{(5-nitro-2-furanyl)methylene}hydrazide (1a) showed equal activity with ceftriaxone against S . aureus . In addition, the MIC values of compounds 1c, 1d and 2a for the same strain were in the range of those reported for ceftriaxone according to NCCLS 1997.

Lancet Infect Dis, 2002 Feb, 2(2), 73 - 85
Resistance of Candida species to antifungal agents: molecular mechanisms and clinical consequences; Sanglard D et al.; Candida albicans and related species pathogenic for man become resistant to antifungal agents, in particular triazole compounds, by expression of efflux pumps that reduce drug accumulation, alteration of the structure or concentration of antifungal target proteins, and alteration of membrane sterol composition . The clinical consequences of antifungal resistance can be seen in treatment failures in patients and in changes in the prevalences of Candida species causing disease . These effects were seen unequivocally in HIV-infected patients with oropharyngeal candida infections, but their incidence has decreased dramatically with the introduction of highly active antiretroviral therapy . The evidence for similar emergence of antifungal-resistant yeast strains and species in other types of candida infections is confounded by non-standardised susceptibility testing methods and definitions of a resistant fungal isolate . Recent large-scale surveys of yeasts isolated from blood cultures, based on standardised methodology and resistance definitions, do not support the view that antifungal resistance in pathogenic yeasts constitutes a significant or growing therapeutic problem.

Eur J Med Chem, 2002 Mar, 37(3), 237 - 53
(2-Arylhydrazonomethyl)-substituted xanthones as antimycotics: synthesis and fungistatic activity against Candida species; Moreau S et al.; A series of arylhydrazones derived from various 6,8-diacetoxy- or 6,8-dihydroxy-9-oxo-9H-xanthene carboxaldehydes were synthesized and evaluated for their in vitro antifungal properties against two human pathogenic yeasts (Candida albicans and C . krusei) according to a diffusion method . The activity was strongly dependent from the position of the (1-arylhydrazinyl-2-ylidene)methyl chain in the xanthone molecular skeleton . Compounds having the nitrogen side chain in the 4-position, with a further halogen substitution on the terminal phenyl ring showed fungistatic effects . Within this series, the 4-fluorophenylhydrazinyl derivative 13g exhibited the highest activity, particularly against C . krusei, with a greater efficacy than that of econazole, used as reference.

Folia Microbiol (Praha), 2001, 46(5), 453 - 7
Electrophoretic karyotyping of Candida albicans strains isolated from premature infants and hospital personnel in a neonatal intensive care unit; Dorko E et al.; Electrophoretic karyotyping was used to compare DNA probes of yeasts isolated from blood of preterm neonates (n = 66) in a neonatal intensive care unit (NICU) and from the hands of healthy hospital personnel (n = 10) . The yeasts were identified as Candida albicans using standard laboratory methods . DNA was extracted from yeasts and isolation of identical DNA strains from the pairs nurse-neonate suggested that one nurse transmitted one yeast strain by her hands to three neonates . Four neonates harbored two identical strains originating from two nurses, i.e . each nurse transmitted the same strain to two neonates . In the additional 7 cases transmission of 1 yeast strain by 1 nurse to 1 neonate was observed . Our data suggest that nonperinatal nosocomial transmission of C . albicans occurs in neonates, possibly via cross-contamination being transferred on hands of health care workers . The importance of careful hand washing of staff (health care workers) and other infection-control procedures (to prevent the nosocomial transmission of pathogens in the NICU environment) is emphasized.

Immunology, 2001 Dec, 104(4), 455 - 61
Human immunodeficiency virus type 1 Tat binds to Candida albicans, inducing hyphae but augmenting phagocytosis in vitro; Gruber A et al.; Tat, the human immunodeficiency virus type 1 (HIV-1) transactivating protein, binds through its RGD-motif to human integrin receptors . Candida albicans, the commonest cause of mucosal candidiasis in subjects infected with HIV-1, also possesses RGD-binding capacity . The present study reveals that Tat binds to C . albicans but not to C . tropicalis . Tat binding was markedly reduced by laminin and to a lesser extent by a complement C3 peptide containing the RGD motif, but not by a control peptide . The outgrowth of C . albicans was accelerated following binding of Tat, but phagocytosis of opsonized C . albicans was also increased after Tat binding . Thus, Tat binding promotes fungal virulence by inducing hyphae but may also reduce it by augmenting phagocytosis . The net effect of Tat in vivo is difficult to judge but in view of the many disease-promoting effects of Tat we propose that accelerating the formation of hyphae dominates over the augmentation of phagocytosis.

Immunology, 2001 Dec, 104(4), 447 - 54
Nitric oxide-enhanced resistance to oral candidiasis; Elahi S et al.; A murine model of oral candidiasis was used to show that nitric oxide (NO) is involved in host resistance to infection with Candida albicans in infection-'resistant' BALB/c and infection-'prone' DBA/2 mice . Following infection, increased NO production was detected in saliva . Postinfection samples of saliva inhibited the growth of yeast in vitro . Treatment with NG-monomethyl-L-arginine (MMLA), an inhibitor of NO synthesis, led to reduced NO production, which correlated with an increase in C . albicans growth . Reduction in NO production following MMLA treatment correlated with an abrogation of interleukin-4 (IL-4), but not interferon-gamma (IFN-gamma), mRNA gene expression in regional lymph node cells . Down-regulation of IL-4 production was accompanied with an increase in IFN-gamma production in infection-'prone' DBA/2 mice . There was a functional relationship between IL-4 and NO production in that mice treated with anti-IL-4 monoclonal antibody showed a marked inhibition of NO production in saliva and in culture of cervical lymph node cells stimulated with C . albicans antigen . The results support previous conclusions that IL-4 is associated with resistance to oral candidiasis and suggest that NO is involved in controlling colonization of the oral mucosal surface with C . albicans.

Antimicrob Agents Chemother, 2002 Apr, 46(4), 1153 - 5
Lack of catheter infection by the efg1/efg1 cph1/cph1 double-null mutant, a Candida albicans strain that is defective in filamentous growth; Lewis RE et al.; The molecular controls regulating the successful colonization of Candida albicans on foreign materials are not known . Here we show that a mutant C . albicans strain defective in filamentous growth and lacking the transcription factors Efg1p and Cph1p has a profoundly deficient potential for colonizing on polyurethane catheters.

Antimicrob Agents Chemother, 2002 Apr, 46(4), 1046 - 52
In vitro low-level resistance to azoles in Candida albicans is associated with changes in membrane lipid fluidity and asymmetry; Kohli A et al.; The present study tracks the development of low-level azole resistance in in vitro fluconazole-adapted strains of Candida albicans, which were obtained by serially passaging a fluconazole-susceptible dose-dependent strain, YO1-16 (fluconazole MIC, 16 microg ml(-1)) in increasing concentrations of fluconazole, resulting in strains YO1-32 (fluconazole MIC, 32 microg ml(-1)) and YO1-64 (MIC, 64 microg ml(-1)) . We show that acquired resistance to fluconazole in this series of isolates is not a random process but is a gradually evolved complex phenomenon that involves multiple changes, which included the overexpression of ABC transporter genes, e.g., CDR1 and CDR2, and the azole target enzyme, ERG11 . The sequential rise in fluconazole MICs in these isolates was also accompanied by cross-resistance to other azoles and terbinafine . Interestingly, fluorescent polarization measurements performed by using the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene revealed that there was a gradual increase in membrane fluidity of adapted strains . The increase in fluidity was reflected by observed change in membrane order, which was considerably decreased (decrease in fluorescence polarization values, P value) in the adapted strain (P value of 0.1 in YO1-64, compared to 0.19 in the YO1-16 strain) . The phospholipid composition of the adapted strain was not significantly altered; however, ergosterol content was reduced in YO1-64 from that in the YO1-16 strain . The asymmetrical distribution of phosphatidylethanolamine (PE) between two monolayers of plasma membrane was also changed, with PE becoming more exposed to the outer monolayer in the YO1-64 strain . The results of the present study suggest for the first time that changes in the status of membrane lipid phase and asymmetry could contribute to azole resistance in C . albicans.

Antimicrob Agents Chemother, 2002 Apr, 46(4), 947 - 57
Candida albicans sterol C-14 reductase, encoded by the ERG24 gene, as a potential antifungal target site; Jia N et al.; The incidence of fungal infections has increased dramatically, which has necessitated additional and prolonged use of the available antifungal agents . Increased resistance to the commonly used antifungal agents, primarily the azoles, has been reported, thus necessitating the discovery and development of compounds that would be effective against the major human fungal pathogens . The sterol biosynthetic pathway has proved to be a fertile area for antifungal development, and steps which might provide good targets for novel antifungal development remain . The sterol C-14 reductase, encoded by the ERG24 gene, could be an effective target for drug development since the morpholine antifungals, inhibitors of Erg24p, have been successful in agricultural applications . The ERG24 gene of Candida albicans has been isolated by complementation of a Saccharomyces cerevisiae erg24 mutant . Both copies of the C . albicans ERG24 gene have been disrupted by using short homologous regions of the ERG24 gene flanking a selectable marker . Unlike S . cerevisiae, the C . albicans ERG24 gene was not required for growth, but erg24 mutants showed several altered phenotypes . They were demonstrated to be slowly growing, with doubling times at least twice that of the wild type . They were also shown to be significantly more sensitive to an allylamine antifungal and to selected cellular inhibitors including cycloheximide, cerulenin, fluphenazine, and brefeldin A . The erg24 mutants were also slightly resistant to the azoles . Most importantly, erg24 mutants were shown to be significantly less pathogenic in a mouse model system and failed to produce germ tubes upon incubation in human serum . On the basis of these characteristics, inhibitors of Erg24p would be effective against C . albicans.

J Oral Pathol Med, 2002 Feb, 31(2), 99 - 105
Prevalence of fluconazole-resistant strains of Candida albicans in otherwise healthy outpatients; Haberland-Carrodeguas C et al.; BACKGROUND: In contrast to the immunosuppressed patient population, the prevalence of fluconazole-resistant strains of Candida albicans among healthy individuals has not been extensively studied . METHODS: Candida species were cultured form 50 healthy outpatients with clinical signs of oral candidiasis . Following one week of the recommended fluconazole regimen, post-treatment cultures were obtained . Both pre- and post-treatment yeasts were identified and in vitro susceptibility testing was performed using the NCCLS M-27A method . Strains were further differentiated using established cDNA probes . RESULTS: Forty-four patients (88%) had positive C.albicans cultures prior to treatment . Antifungal susceptibility testing of these strains demonstrated no in vitro resistance to fluconazole . At post-treatment evaluation, eight patients (18%) had persistent signs of infection and 10 patients (23%) had positive Candida sp . cultures despite no clinical signs of infection . DNA analysis confirmed that the same C . albicans strain was present both in the pre-treatment and the post-treatment cultures . CONCLUSIONS: Our results showed that the presence of fluconazole-resistant strains of C.albicans does not appear to be prevalent among healthy outpatients furthermore, in vitro antifungal susceptibility testing does not always predict successful therapy in these patients.

Infect Immun, 2002 Apr, 70(4), 1772 - 82
Disruption of the Candida albicans TPS2 gene encoding trehalose-6-phosphate phosphatase decreases infectivity without affecting hypha formation; Van Dijck P et al.; Deletion of trehalose-6-phosphate phosphatase, encoded by TPS2, in Saccharomyces cerevisiae results in accumulation of trehalose-6-phosphate (Tre6P) instead of trehalose under stress conditions . Since trehalose is an important stress protectant and Tre6P accumulation is toxic, we have investigated whether Tre6P phosphatase could be a useful target for antifungals in Candida albicans . We have cloned the C . albicans TPS2 (CaTPS2) gene and constructed heterozygous and homozygous deletion strains . As in S . cerevisiae, complete inactivation of Tre6P phosphatase in C . albicans results in 50-fold hyperaccumulation of Tre6P, thermosensitivity, and rapid death of the cells after a few hours at 44 degrees C . As opposed to inactivation of Tre6P synthase by deletion of CaTPS1, deletion of CaTPS2 does not affect hypha formation on a solid glucose-containing medium . In spite of this, virulence of the homozygous deletion mutant is strongly reduced in a mouse model of systemic infection . The pathogenicity of the heterozygous deletion mutant is similar to that of the wild-type strain . CaTPS2 is a new example of a gene not required for growth under standard conditions but required for pathogenicity in a host . Our results suggest that Tre6P phosphatase may serve as a potential target for antifungal drugs . Neither Tre6P phosphatase nor its substrate is present in mammals, and assay of the enzymes is simple and easily automated for high-throughput screening.

Food Chem Toxicol, 2002 Apr, 40(4), 535 - 43
Toxicity of Australian essential oil Backhousia citriodora (Lemon myrtle) . Part 1 . Antimicrobial activity and in vitro cytotoxicity; Hayes AJ et al.; The antimicrobial and toxicological properties of the Australian essential oil, lemon myrtle, (Backhousia citriodora) were investigated . Lemon myrtle oil was shown to possess significant antimicrobial activity against the organisms Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, methicillin-resistant S . aureus (MRSA), Aspergillus niger, Klebsiella pneumoniae and Propionibacterium acnes comparable to its major component-citral . An in vitro toxicological study based on the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) cytotoxicity assay was performed . In vitro cytotoxicity testing indicated that both lemon myrtle oil and citral had a very toxic effect against human cell lines: HepG2 (a hepatocarcinoma-derived cell line); F1-73 (a fibroblast cell line derived from normal skin) and primary cell cultures of human skin fibroblasts . Cytotoxicity IC50 (50% inhibitory concentration) values ranged from 0.008 to 0.014% (w/v) at 4 h to 0.003-0.012% (w/v) at 24 h of exposure . The no-observed-adverse-effect level (NOAEL) for lemon myrtle oil was calculated as 0.5 mg/l at 24 h exposure and the RfD (reference dose) was determined as 0.01 mg/l . A product containing 1% lemon myrtle oil was found to be low in toxicity and could potentially be used in the formulation of topical antimicrobial products.

J Pept Res, 2001 Nov, 58(5), 349 - 56
Antimicrobial peptides from the skin of the Japanese mountain brown frog, Rana ornativentris; Kim JB et al.; Six peptides with antimicrobial activity were isolated from an extract of freeze-dried skin of the Japanese mountain brown frog Rana ornativentris . Two structurally related peptides (brevinin-20a GLFNVFKGALKTAGKHVAGSLLNQLKCKVSGGC, 11 nmol/g dried tissue, and brevinin-20b GIFNVFKGALKTAGKHVAGSLLNQLKCKVSGEC, 170 nmol/g) belong to the brevinin-2 family, previously identified in Asian and European, but not North American, Ranid frogs . Four peptides (temporin-10a FLPLLASLFSRLL.NH2, 13 nmol/g; temporin-10b FLPLIGKILGTI L.NH2, 350 nmol/g; temporin-10c FLPLLASLFSRLF.NH2, 14 nmol/g; and temporin-10d FLPLLASLFSGLF.NH2, 8 nmol/g) are members of the temporin family first identified in the European common frog Rana temporaria but also found in the skins of North American Ranids . The brevinin-2 peptides showed broad-spectrum activity against the gram-positive bacterium, Staphylococcus aureus, the gram-negative bacterium, Escherichia coli and the yeast Candida albicans, whereas the temporins showed potent activity only against S . aureus . The brevinins and temporins belong to the class of cationic antimicrobial peptides that adopt an amphipathic alpha-helical conformation but it is significant that temporin-10d, which lacks a basic amino acid residue, is still active against S . aureus (minimum inhibitory concentration=13 microM compared with 2 microM for temporin-10a) . This suggests that strong electrostatic interaction between the peptide and the negatively charged phospholipids of the cell membrane is not an absolute prerequisite for antimicrobial activity.

Ostomy Wound Manage, 2000 Oct, 46(10), 28 - 34
A comparative study of the effects of UVC irradiation on select procaryotic and eucaryotic wound pathogens; Sullivan PK et al.; Managing wounds infected with a mixture of several types of microorganisms such as bacteria (procaryotes) and fungi (eucaryotes) is a challenging clinical situation . The purpose of this study was to determine the effectiveness of ultraviolet light (UVC) in eradicating select procaryotic and eucaryotic organisms, in both pure culture and mixed cultures in vitro . Five replications of each organism or mixture of organisms (10(6) organisms/mL singly or 10(15) organisms/mL mixed culture) were plated . The cultures were treated with a UVC light 1 inch from the surface . Irradiation times were 0, 2, 3, 4, 5, 15, 30, 45, 60, 90, 120, and 180 seconds . Bacterial cultures were incubated and colony counts performed . Upon exposure to UVC, a 99.9% kill rate was obtained at 3 to 5 seconds for the procaryotic organisms (Pseudomonas aeruginosa and Mycobacterium abscessus) tested . However, 15 to 30 seconds of UVC treatment was required to obtain 99.9% kill of the eucaryotic organisms (Candida albicans, Aspergillus fumigatus) tested . This study demonstrates a decreasing sensitivity of evolutionarily more complex organisms to UVC . This study also provides further evidence that short exposure times to UVC are detrimental to procaryotic and simple unicellular eucaryotic organisms while sparing more complex multicellular organisms.

J Bacteriol, 2002 Apr, 184(7), 2058 - 61
Conserved serine/threonine kinase encoded by CBK1 regulates expression of several hypha-associated transcripts and genes encoding cell wall proteins in Candida albicans; McNemar MD et al.; The opportunistic fungal pathogen, Candida albicans, is reported to have several potential virulence factors . A potentially significant factor is the ability to undergo morphological transition from yeast to hypha . This alteration of form is accompanied by many changes within the cell, including alterations in gene expression and cell wall composition . We have isolated a gene that encodes a highly conserved serine/threonine kinase that appears to be involved in the regulation of proteins associated with the cell wall . We have assigned the designation CBK1 (cell wall biosynthesis kinase 1) to this gene . Mutants lacking CBK1 form large aggregates of round cells under all growth conditions and lack the ability to undergo morphological differentiation . Additionally, these mutants show an altered pattern of expression of several transcripts encoding proteins associated with the cell wall . The results suggest that the kinase encoded by CBK1 plays a general role in the maintenance and alteration of the cell wall of C . albicans in all morphologies.

Chest, 2002 Mar, 121(3), 782 - 8
Serologic IgE immune responses against Aspergillus fumigatus and Candida albicans in patients with cystic fibrosis; Maiz L et al.; OBJECTIVES: The objectives of this study were to determine the prevalence of Aspergillus fumigatus and Candida albicans in the sputa of patients with cystic fibrosis (CF), to assess serologic IgE responses of these patients to the presence of fungi in the sputum, to evaluate what effect this may have on clinical status, and to determine how the above-mentioned factors relate to allergic bronchopulmonary aspergillosis (ABPA) . PATIENTS: Seventy-six CF patients (40 male and 36 female patients; age, 15.3 plus minus 8.7 years {mean plus minus SD}) were studied . Measurements and results: A total of 1,239 sputum samples from 66 patients were cultured for fungi . A fumigatus was grown in 256 sputum specimens (20.7%), and C albicans was grown in 588 sputum samples (47.5%) . Forty patients (60.6%) had at least one positive culture finding for A fumigatus, and 58 patients (87.9%) had at least one positive culture finding for C albicans . Forty-nine patients (64.5%) were sensitized to A fumigatus, and 20 patients (26.7%) were sensitized to C albicans . No correlation was found between the finding of A fumigatus in sputum and IgE to A fumigatus . Only patients who had at least one positive culture finding for C albicans had IgE to C albicans develop . Lung function values and chest radiograph scores were not significantly lower in patients sensitized to either A fumigatus or C albicans as compared to nonsensitized patients . Of the 20 patients sensitized to C albicans, 10 patients had confirmed ABPA and 10 patients had some immunologic characteristics of ABPA . CONCLUSIONS: A high prevalence of colonization and sensitization to A fumigatus and C albicans in CF patients was observed . The sensitization to these fungi was not related to the clinical severity . IgE to C albicans may be an immunologic marker related to the development of ABPA in patients with CF.

Eur J Cardiothorac Surg, 2002 Mar, 21(3), 591 - 2
Pericardial constriction caused by Candida albicans; Benedik J Jr et al.; An uncommon occurrence of constrictive pericarditis caused by Candida albicans and its treatment by successful pericardectomy and epicardectomy are described . For pathogen detection, both cultivation and molecular diagnostics were used . The speed and reliability of molecular diagnostics using polymerase chain reaction make this method a powerful tool for pathogen detection in any clinical specimen.

Med Oral, 2002 Mar-Apr, 7(2), 121 - 9
Prevalence of diabetes mellitus amongst oral lichen planus patients . Clinical and pathological characteristics; Romero MA et al.; OBJECTIVES: The aim of this study has been to define the prevalence of DM (considered according to the ADA-97 criteria) in OLP patients and also to investigate the existence of clinical and pathological differences between OLP patients with or without DM . STUDY DESIGN: Sixty-two patients suffering from OLP according to clinical and pathological criteria were selected to enter the study and classified after the DM diagnosis guidelines suggested by The American Diabetes Association (ADA-1997) . The variables considered for each patient were: age, sex, clinical presentation, extension of the lesions, location of the lesions, number of locations, Candida albicans colonization, and density of subepithelial inflammatory infiltrate . RESULTS: Up to 27.4% of OLP cases were associated to type 2 DM (DM2) and 17.7% were related to an impaired fasting glucose (IFG) . The mean age of the DM2-associated OLP group was significantly higher than the non-diabetic group's . No significant differences could be observed in terms of clinical and pathological features between diabetic and non-diabetic OLP patients . CONCLUSION: The high prevalence of hydrocarbonate metabolism disorders observed in OLP patients justify the use of FPG for screening of DM2 in these patients.

Mol Microbiol, 2001 Dec, 42(5), 1243 - 57
Distinct and redundant roles of the two protein kinase A isoforms Tpk1p and Tpk2p in morphogenesis and growth of Candida albicans; Bockmuhl DP et al.; TPK1 and TPK2 encode both isoforms of protein kinase A (PKA) catalytic subunits in Candida albicans . Mutants lacking both TPK1 alleles showed defective hyphal morphogenesis on solid inducing media, whereas in liquid hypha, formation was affected slightly . In contrast, tpk2 mutants were only partially morphogenesis defective on solid media, whereas a strong block was observed in liquid . In addition, the yeast forms of tpk2-- but not tpk1-- mutants were completely deficient in invading agar . Because Tpk1p and Tpk2p differ in their N-terminal domains of approximately 80--90 amino acids, while the catalytic portions are highly homologous, the functions of hybrid Tpk proteins with exchanged N-terminal domains were tested . The results demonstrate that the catalytic portions mediate Tpk protein specificities with regard to filamentation, whereas agar invasion is mediated by the N-terminal domain of Tpk2p . Homozygous tpk1 and tpk2 mutants grew normally; however, a tpk2 mutant strain containing a single regulatable TPK1 allele (PCK1p-TPK1) at low expression levels was severely growth defective . It was completely blocked in hyphal morphogenesis and was stress resistant to high osmolarities or temperatures . Thus, both Tpk isoforms in C . albicans share growth functions but, unlike Saccharomyces cerevisiae isoforms, they have positive, specific roles in filament formation in different environments.

J Immunol, 2002 Mar 15, 168(6), 2904 - 13
Dendritic cells pulsed with fungal RNA induce protective immunity to Candida albicans in hematopoietic transplantation; Bacci A et al.; Immature myeloid dendritic cells (DC) phagocytose yeasts and hyphae of the fungus Candida albicans and induce different Th cell responses to the fungus . Ingestion of yeasts activates DC for production of IL-12 and Th1 priming, while ingestion of hyphae induces IL-4 production and Th2 priming . In vivo, generation of antifungal protective immunity is induced upon injection of DC ex vivo pulsed with Candida yeasts but not hyphae . In the present study we sought to determine the functional activity of DC transfected with yeast or hyphal RNA . It was found that DC, from either spleens or bone marrow, transfected with yeast, but not hyphal, RNA 1) express fungal mannoproteins on their surface; 2) undergo functional maturation, as revealed by the up-regulated expression of MHC class II Ags and costimulatory molecules; 3) produce IL-12 but no IL-4; 4) are capable of inducing Th1-dependent antifungal resistance when delivered s.c . in vivo in nontransplanted mice; and 5) provide protection against the fungus in allogeneic bone marrow-transplanted mice, by accelerating the functional recovery of Candida-specific IFN-gamma-producing CD4(+) donor lymphocytes . These results indicate the efficacy of DC pulsed with Candida yeasts or yeast RNA as fungal vaccines and point to the potential use of RNA-transfected DC as anti-infective vaccines in conditions that negate the use of attenuated microorganisms or in the case of poor availability of protective Ags.

J Clin Microbiol, 2002 Mar, 40(3), 852 - 6
Trends in antifungal susceptibility of Candida spp . isolated from pediatric and adult patients with bloodstream infections: SENTRY Antimicrobial Surveillance Program, 1997 to 2000; Pfaller MA et al.; From 1 January 1997 through 31 December 2000, 2,047 bloodstream infections (BSIs) due to Candida spp . were reported from hospitals in the United States, Canada, Latin America, and Europe participating in the SENTRY Antifungal Surveillance Program . Among individuals in four age groups (< or =1, 2 to 15, 16 to 64, and > or =65 years) Candida albicans was the most common species, causing 60, 55, 55, and 50% of infections, respectively . C . glabrata caused 17 to 23% of BSIs in those ages 16 to 64 and > or = 65 years, whereas it caused only 3% of BSIs in the individuals in the two younger age groups (P < 0.001) . C . parapsilosis (which caused 21 to 24% of BSIs) and C . tropicalis (which caused 7 to 10% of BSIs) were more common than C . glabrata in individuals ages < or =1 year and 2 to 15 years . Isolates of Candida spp . showed a trend of decreasing susceptibility to fluconazole, itraconazole, and amphotericin B with increasing patient age (P < or = 0.01) . None of the C . glabrata isolates from individuals < or =1 year old were resistant to fluconazole, whereas they made up 5 to 9% of isolates from individuals ages 16 to 64 and > or =65 years . Isolates of C . tropicalis from patients < or =1 year old were more susceptible to flucytosine (MIC at which 90% of isolates are inhibited {MIC(90)}, 0.5 microg/ml; 0% resistant isolates) than those from patients > or =65 years old (MIC(90), 32 microg/ml; 11% resistant isolates) . The investigational triazoles posaconazole, ravuconazole, and voriconazole were all highly active against all species of Candida from individuals in all age groups . These data demonstrate differences in the species distributions of pathogens and differences in antifungal resistance among isolates from individuals in the pediatric and adult age groups . Ongoing surveillance will enhance efforts to limit the extent of antifungal resistance in individuals in various age groups.

J Clin Microbiol, 2002 Mar, 40(3), 826 - 36
Ca3 fingerprinting of Candida albicans isolates from human immunodeficiency virus-positive and healthy individuals reveals a new clade in South Africa; Blignaut E et al.; To examine the question of strain specificity in oropharyngeal candidiasis associated with human immunodeficiency virus (HIV) infection, oral samples were collected from 1,196 HIV-positive black South Africans visiting three clinics and 249 Candida albicans isolates were selected for DNA fingerprinting with the complex DNA fingerprinting probe Ca3 . A total of 66 C . albicans isolates from healthy black South Africans and 46 from healthy white South Africans were also DNA fingerprinted as controls . Using DENDRON software, a cluster analysis was performed and the identified groups were compared to a test set of isolates from the United States in which three genetic groups (I, II, and III) were previously identified by a variety of genetic fingerprinting methods . All of the characterized South African collections (three from HIV-positive black persons, two from healthy black persons, and one from healthy white persons) included group I, II, and III isolates . In addition, all South African collections included a fourth group (group SA) completely absent in the U.S . collection . The proportion of group SA isolates in HIV-positive and healthy black South Africans was 53% in both cases . The proportion in healthy white South Africans was 33% . In a comparison of HIV-positive patients with and without oropharyngeal symptoms of infection, the same proportions of group I, II, III, and SA isolates were obtained, indicating no shift to a particular group on infection . However, by virtue of its predominance as a commensal and in infections, group SA must be considered the most successful in South Africa . Why group SA isolates represent 53 and 33% of colonizing strains in black and white South Africans and are absent in the U.S . collection represents an interesting epidemiological question.

Eur J Pharm Biopharm, 2002 Mar, 53(2), 233 - 9
Formulation and in vivo evaluation of chlorhexidine buccal tablets prepared using drug-loaded chitosan microspheres; Giunchedi P et al.; This investigation deals with the development of buccal formulations (tablets) based on chitosan microspheres containing chlorhexidine diacetate . The microparticles were prepared by a spray-drying technique, their morphological characteristics were studied by scanning electron microscopy and the in vitro release behaviour was investigated in pH 7.0 USP buffer . Chlorhexidine in the chitosan microspheres dissolves more quickly in vitro than does chlorhexidine powder . The anti-microbial activity of the microparticles was investigated as minimum inhibitory concentration, minimum bacterial concentration and killing time . The loading of chlorhexidine into chitosan is able to maintain or improve the anti-microbial activity of the drug . The improvement is particularly high against Candida albicans . This is important for a formulation whose potential use is against buccal infections . Drug-empty microparticles have an anti-microbial activity due to the polymer itself . Buccal tablets were prepared by direct compression of the microparticles with mannitol alone or with sodium alginate . After their in vivo administration the determination of chlorhexidine in saliva showed the capacity of these formulations to give a prolonged release of the drug in the buccal cavity.

Zhonghua Shao Shang Za Zhi, 2000 Apr, 16(2), 108 - 10
{The relationship between postburn gene expression of modulators in gut associated lymph tissue and the change in IgA plasma cells}; Bai X et al.; OBJECTIVE: To investigate the relationship between the postburn change in IgA plasma cells and those of IL- 4 and IL-6 in gut associated lymph tissue (GALT) . METHODS: One hundred and twenty -- five SPF mice were enrolled in the study and randomly divided into three groups -- normal control (A), microbial inoculation (B) and microbial inoculation followed by burn injury . (C) In groups Band C, the mice were gavaged with candida albicans, and the mice in group C, were inflicted with 20% TBSA III degree burn at the 14th day after inoculation . The mice were sacrificed thereafter on 1, 2 and 3 postburn days and the samples were obtained aseptically . The number of candida albicans adhering to intestinal mucosal membrane was counted . The number of IgA plasma cells in lamina propria was determined by immunohistochemical staining . The gene expression of IL-4 in Peyer patch was detected by dothybridization and that of IL-6 in lamina propria was determined by insitu hybridization . RESULTS: (1) The postburn gene expression of IL-4 in Peyer patch decreased, but the IL-6 level in intestinal lamina propria increased . (2) The number of IgA plasma cells in lamina propria decreased significantly, but the adherence of candida albicans to intestinal mucous membrane increased obviously . CONCLUSION: The decrement of postburn gene expression of IL-4 in Peyer patch might lead to IgA plasma cell decrease, resulting in the increase of candida albicans adhering to intestinal mucosal membrane.

J Appl Microbiol, 2002, 92(3), 549 - 55
A quantitative study of the survival of two species of Candida on porous and non-porous environmental surfaces and hands; Traore O et al.; AIMS: In spite of the importance of many species of Candida as human pathogens, little is known about their ability to survive on animate and inanimate surfaces . Such information is essential in understanding the vehicles and modes of their spread, and in designing proper infection control strategies against them . The aim of this study was to generate comparative quantitative data in this regard . METHODS AND RESULTS: The survival of one clinical isolate each of Candida albicans and C . parapsilosis on two types of hard inanimate surfaces (glass and stainless steel) and two types of fabrics (100% cotton and a blend of 50% cotton and 50% polyester) was evaluated under ambient conditions (air temperature 22 +/- 2 degrees C; relative humidity 45-62%) using quantitative test protocols . The survival of C . albicans was also assessed on human skin, using the fingerpads of adult volunteers as carriers . Each carrier surface received 10 microl of the test suspension containing a soil load to simulate body fluids . When dried on glass and stainless steel carriers, C . albicans and C . parapsilosis remained viable for at least three and 14 days, respectively . Both species could survive for at least 14 days on both types of fabric . On the skin, 20% of the viable C . albicans remained detectable one hour post-inoculation . SIGNIFICANCE AND IMPACT OF THE STUDY: This quantitative and comparative study demonstrated the potential for, and differences in the ability of clinically significant species of Candida to remain viable on porous and non-porous inanimate surfaces as well as on human hands . These results should help in understanding the epidemiology of nosocomial infections due to Candida, and in designing better prevention and control strategies against them.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Sep-Oct, (5), 30 - 4
{Experimental reactivity and antigenic activity of the bacterial-fungal antigen complexes}; Kazhina MV et al.; The immunogenic and reactogenic properties of monovaccines prepared from Staphylococcus aureus, S . epidermidis and Candida albicans, as well as those of a associated polyvaccine prepared from these infective agents, were experimentally studied on rabbits . The monovaccines and the associated bacterial-fungal vaccines were found to be safe and faintly reactogenic; in the blood serum of vaccinated rabbits a growth in the titer of agglutinins and its preservation at a high level for 4 months were noted.

Yeast, 2002 Mar 15, 19(4), 341 - 9
Inhibitory effect of nikkomycin Z on chitin synthases in Candida albicans; Kim MK et al.; Nikkomycin Z is a competitive inhibitor of chitin synthases in fungi . It has been reported that it inhibits chitin synthases (Chs) 1 and 3, but not 2, of Saccharomyces cerevisiae . In our study, we found that: (a) nikkomycin Z inhibited all three Chs isozymes of Candida albicans (CaChs) . The IC(50) value for CaChs1 is 15microM, for CaChs2 0.8microM, and for CaChs3 13microM; (b) nikkomycin Z inhibits vegetative growth of C . albicans differently in different growth media; growth inhibition was observed on Spider and corn meal agar plate, but not on Lee's plate; (c) growth inhibition by nikkomycin Z accompanied by the absence of septum and cell wall chitin, which in turn brought about cell lysis . Nikkomycin Z did not lyse cells in Lee's media and lysis was partially prevented in the presence of sorbitol as an osmostabilizer in Spider medium . Therefore, nikkomycin Z prevented the formation of septum and cell wall chitin by inhibiting chitin synthase activities in a growth medium-dependent manner .

Yeast, 2002 Mar 15, 19(4), 329 - 40
Siderophore uptake by Candida albicans: effect of serum treatment and comparison with Saccharomyces cerevisiae; Lesuisse E et al.; Iron uptake systems often function as virulence factors in pathogenic organisms . Candida albicans is a fungal pathogen that infects immunocompromised hosts, such as AIDS patients or granulocytopenic bone marrow transplant recipients . Here we show that iron uptake from siderophores occurs in C . albicans and is mediated by one or more high-affinity transport systems . Iron carried on ferrioxamine B, triacethyl-fusarinine, ferrichrome, or ferricrocin was actively taken up via a high-affinity mechanism . The kinetic parameters of uptake were similar to those found in S . cerevisiae . Furthermore, for ferrichrome and ferrioxamine B, cellular uptake of fluorescent analogues was observed . In C . albicans, iron uptake from siderophores was regulated by iron availability, with iron deprivation inducing uptake . Serum exposure, which induces a morphogenic shift from yeast to filamentous forms known to be required for virulence, also resulted in induction of iron transport from ferrichrome-type siderophores . In a tup1/tup1 strain which grows constitutively in the filamentous form, iron transport was derepressed for all siderophores tested . The genes mediating uptake and utilization of iron from siderophores in C . albicans have not been identified; however, the transcript abundance for CaSIT1 was regulated in a manner consistent with the pattern of iron uptake from ferrichrome-type siderophores . Furthermore, CaSIT1 overexpression in S . cerevisiae resulted in inhibited siderophore iron uptake, suggesting that the expressed protein may interact with proteins of S . cerevisiae involved in iron uptake from siderophores . In summary, iron uptake from ferrichrome-type siderophores was induced in filamentous C . albicans, and a potential role of this iron acquisition system in pathogenicity should be considered .

Yeast, 2002 Mar 15, 19(4), 303 - 18
ABC transporters Cdr1p, Cdr2p and Cdr3p of a human pathogen Candida albicans are general phospholipid translocators; Smriti et al.; We have used fluorescent 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD)-tagged phospholipid analogues, NBD-PE (phosphatidylethanolamine), NBD-PC (phosphatidylcholine) and NBD-PS (phosphatidylserine), to demonstrate that Cdr1p and its other homologues, Cdr2p and Cdr3p, belonging to the ATP-binding cassette (ABC) superfamily behave as general phospholipid translocators . Interestingly, CDR1 and CDR2, whose overexpression leads to azole resistance in C . albicans, elicit in-to-out transbilayer phospholipid movement, while CDR3, which is not involved in drug resistance, carries out-to-in translocation of phospholipids between the two monolayers of plasma membrane . Cdr1p, Cdr2p and Cdr3p could be further distinguished on the basis of their sensitivities to different inhibitors . For example, the in-to-out activity associated with Cdr1p and Cdr2p is energy-dependent and sensitive to sulphydryl blocking agents such as N-ethylmaleimide (NEM) and cytoskeleton disrupting agent cytochalasin E, while Cdr3p-associated out-to-in activity is energy-dependent but insensitive to NEM and cytochalasin E . We found that certain drugs, such as fluconazole, cycloheximide and miconazole, to which Cdr1p confers resistance could also affect in-to-out transbilayer movement of NBD-PE, while the same drugs had no effect on Cdr3p-mediated out-to-in translocation of NBD-PE . The ineffectiveness of these drugs to affect Cdr3p mediated out-to-in phospholipid translocation further confirms the inherent difference in the directionality of phospholipid translocation between these pumps . Notwithstanding the role of some of the Cdrps in drug resistance, this study clearly demonstrates that these ABC transporters of C . albicans are phospholipid translocators and this function could represent one of the physiological functions of such large family of proteins .

J Biomed Mater Res, 2002, 63(2), 122 - 8
Surface modification of an experimental silicone rubber aimed at reducing initial candidal adhesion; Price C et al.; Silicone rubber, which is a widely used biomaterial, is often used to make soft liners for permanent denture . Colonization of denture soft lining materials by Candida albicans can result in clinical problems . The aim of this study was to chemically modify the surface of an experimental silicone rubber in order to produce a silicone that was less susceptible to candidal colonization . Surface modification was carried out with the use of argon-plasma bombardment followed by silane treatment, which caused the incorporation of either hydrophilic or hydrophobic functional groups onto the surface . Changes in water contact angles and chemical analysis of the materials with scanning ion mass spectroscopy confirmed surface changes . In vitro assays were carried out using C . albicans to measure levels of adherence to the surface-modified silicone after 1 h . C . albicans exhibited very low adherence to all silane-treated surfaces, whether hydrophobic or hydrophilic . This led to the conclusion that incorporated long-chain functional groups were inhibiting the adherence of the yeast, possibly by the formation of a barrier between the surface of the material and the yeast . In conclusion, silane surface treatment of an experimental silicone rubber has been successful in reducing candidal adherence .

Clin Microbiol Infect, 1996 Aug, 2(1), 36 - 43
Anti-enolase antibodies partially protective against systemic candidiasis in mice; van Deventer HJ et al.; OBJECTIVE: The evaluation of the effect of protective capacities of antibodies directed against the immunodominant enolase antigen of C . albicans on the survival rate of mice with systemic candidiasis . METHODS: Passive transfer of heterologous rabbit antibodies (IRS) directed against a crude Candida albicans extract and homologous mice anti-enolase antibodies (IMS) in a non-acute but lethal systemic Candida model in mice . RESULTS: Protection could only be demonstrated by repeated administration of homologous as well as heterologous immune sera . The protective effect was not due to non-specific, heat-labile, serum factors, since fractionated anti-cytoplasmic rabbit immunoglobulins also gave a decreased mortality rate . These findings could not be ascribed to enhanced opsonization as observed in in vitro opsonization experiments using peritoneal macrophage monolayers . CONCLUSIONS: Anti-enolase antibodies are partially protective against lethal C . albicans infections . The protection was not due to enhanced initial clearance of viable C . albicans from the internal organs, but was presumably the result of as yet unknown effects of antibodies occurring later in the infectious process.

Clin Microbiol Infect, 1995 Feb, 1(2), 124 - 126
Fungemia: An increasing problem in a Danish university hospital 1989 to 1994; Bruun B et al.; OBJECTIVE: To present data on episodes of fungemia in a Danish tertiary-care university hospital admitting all types of patients and to compare the data with previous findings from the same hospital . METHODS: Retrospective identification of episodes of fungemia from 1989 to 1994 and collection of data from computerized files at the Clinical Microbiology Department at Rigshospitalet and the Mycology Reference Laboratory at Statens Seruminstitut . RESULTS: The incidence of fungemia increased gradually from 19 episodes in 1989 to 57 episodes in 1994 . An earlier report from the same hospital showed 20 to 25 episodes of fungemia per year between 1984 and 1988 . Candida albicans was the dominating species during both periods, accounting for 73% of isolates during 1984 to 1988 and 67% during 1989 to 1994 . However, in the hematology department where fluconazole has been used extensively, C . albicans constituted 47% of isolates with Candida krusei and Candida glabrata comprising 25% . CONCLUSIONS: The incidence of fungemia in our tertiary-care hospital has increased threefold from 1989 to 1994 . Candida albicans was the dominating cause of fungemia but, in the hematology department, this yeast accounted for less than half of the isolates during the same time period.

Biochem Biophys Res Commun, 2002 Mar 8, 291(4), 1006 - 13
Antifungal mechanism of an antimicrobial peptide, HP (2--20), derived from N-terminus of Helicobacter pylori ribosomal protein L1 against Candida albicans; Lee DG et al.; The antifungal activity and mechanism of HP (2-20), a peptide derived from the N-terminus sequence of Helicobacter pylori Ribosomal Protein L1 were investigated . HP (2--20) displayed a strong antifungal activity against various fungi, and the antifungal activity was inhibited by Ca(2+) and Mg(2+) ions . In order to investigate the antifungal mechanism(s) of HP (2-20), fluorescence activated flow cytometry was performed . As determined by propidium iodide staining, Candida albicans treated with HP (2-20) showed a higher fluorescence intensity than untreated cells and was similar to melittin-treated cells . The effect on fungal cell membranes was examined by investigating the change in membrane dynamics of C . albicans using 1,6-diphenyl-1,3,5-hexatriene as a membrane probe and by testing the membrane disrupting activity using liposome (PC/PS; 3:1, w/w) and by treating protoplasts of C . albicans with the peptide . The action of peptide against fungal cell membrane was further examined by the potassium-release test, and HP (2-20) was able to increase the amount of K(+) released from the cells . The result suggests that HP (2-20) may exert its antifungal activity by disrupting the structure of cell membrane via pore formation or directly interacts with the lipid bilayers in a salt-dependent manner.

J Infect Dis, 2002 Feb 15, 185(4), 521 - 30 Epub 2002 Jan 31.
Essential role of the Candida albicans transglutaminase substrate, hyphal wall protein 1, in lethal oroesophageal candidiasis in immunodeficient mice; Sundstrom P et al.; Oroesophageal candidiasis is caused by the combined action of fungal virulence factors and host inflammatory responses when protective immunity is absent . Hyphal wall protein 1 (Hwp1) on germ tubes and true hyphae of Candida albicans forms covalent cross-links to buccal epithelial cells in vitro by functioning as a substrate for mammalian transglutaminases . In this study, beige-athymic (bg/bg-nu/nu) or transgenic epsilon 26 mice that have combined natural killer and T cell defects did not succumb to candidiasis after oral administration of C . albicans strains with inactivated HWP1 genes, whereas mice given isogenic strains of C . albicans that had a single copy of HWP1 survived only 2-3 weeks . Illness correlated with extensive alterations of the lingual and esophageal mucosa that were absent in mice given the hwp1/hwp1 mutant . HWP1 is a promising target for development of antifungal drugs for treatment of oroesophageal candidiasis.

J Antimicrob Chemother, 2002 Mar, 49(3), 515 - 24
Heterogeneous mechanisms of azole resistance in Candida albicans clinical isolates from an HIV-infected patient on continuous fluconazole therapy for oropharyngeal candidosis; Martinez M et al.; Molecular mechanisms of azole resistance in Candida albicans include alterations in the target enzyme and increased efflux of drug, but the impact of specific treatment regimens on resistance has not been established . A patient with advanced AIDS was enrolled in a longitudinal study to receive continuous oral fluconazole (FLU) 200 mg/day for the treatment of oropharyngeal candidosis (OPC) . Oral cultures were obtained at time of enrollment, during episodes of OPC and quarterly for surveillance . The patient had five symptomatic relapses on continuous FLU during 43 months . All OPC episodes were successfully treated with increasing doses of FLU although increased FLU MICs were detected for C . albicans isolates with progression of time . DNA-typing techniques demonstrated that resistance developed in a persistent strain of C . albicans . Both FLU-resistant and isogenic isolates with reduced susceptibility were detected in the same clinical samples through multiple episodes . Analysis of molecular mechanisms of resistance revealed overexpression of MDR and CDR genes encoding efflux pumps (but not ERG11) in isolates with decreased FLU susceptibility . In addition, the presence of the G464S amino acid substitution in their lanosterol demethylase, affecting its affinity for FLU, was also detected . However, other isogenic, but FLU-susceptible isolates recovered from the same samples did not harbour the mutation, indicating microevolution of yeast populations within the oral cavity . In this patient, the continuous antifungal pressure exerted by FLU resulted in development of resistance of multifactorial nature . Despite their clonal origin, different subpopulations of C . albicans demonstrated distinct resistance mechanisms, including concomitant presence and absence of functional point mutations in ERG11 genes.

Clin Microbiol Infect, 1998 Feb, 4(2), 82 - 87
Evaluation of the discriminatory power of pulsed-field gel electrophoresis and PCR fingerprinting for epidemiologic typing of Candida species; Voss A et al.; OBJECTIVE: To evaluate the discriminatory power of genotyping methods (PCR fingerprinting and pulsed-field gel electrophoresis) validated for Candida albicans in other Candida species . METHODS: Molecular typing methods are increasingly being applied for studies where the interpretation of data essentially relies on the typing results rather than epidemiologic data . In this situation, the discriminatory power (ability to identify differences among epidemiologically unrelated strains) of the typing method is important in allowing one to draw valid conclusions . By applying PCR fingerprinting, electrophoretic karyotyping, and restriction fragment endonuclease analysis using standard restriction enzymes and primers proven to be useful in previous studies, we evaluated whether the use of multiple genotyping methods is sufficient to delineate known unrelated strains among seven Candida species . RESULTS: All three methods identified individual genotypes for each of the seven Candida species studied . However, optimal strain delineation required the combined use of all three typing methods and was observed only within the small number of C . albicans and C . tropicalis isolates tested in this study . CONCLUSION: Typing assays that are able to delineate a certain Candida species may not be used blindly for other species of that genus . Regarding the limited number of strains tested, further validation of the discriminative power of genotyping methods (including in C . tropicalis) should be done.

Clin Microbiol Infect, 1997 Apr, 3(2), 192 - 197
Invasive candidosis in pediatric patients; Roilides E et al.; OBJECTIVE: To analyze invasive Candida infections in pediatric patients and to examine the outcome of disease . METHODS: An observational prospective study was carried out of microbiologically documented cases of invasive candidosis in neonates, infants and children at Hippokration Hospital from December 1993 to July 1995 . RESULTS: Thirty-nine cases of invasive candidosis (mainly candidemia and candiduria) occurred in 38 patients aged 3 days to 14 years, 21 (54%) having occurred in patients aged <1 month . The incidence was 10.1/1000 admissions in the two neonatal departments versus 1.1/1000 for all other pediatric departments, including oncology (p<0.01) . The most frequent causes of hospitalization were premature birth or perinatal problems and surgery . Thirty-five strains of Candida albicans, 10 of C . parapsilosis and eight of other or unidentified species were isolated . Non-C . albicans isolates, especially C . parapsilosis and C . glabrata, exhibited higher minimal inhibitory concentrations of azoles as compared with C . albicans . Thirteen patients (34%), all candidemic, died within 0--40 days . Untreated patients more frequently had positive cultures up to time of death (p<0.0001) and died (p=0.0006) . CONCLUSIONS: Invasive candidosis is frequent in pediatric patients, especially premature neonates, and is associated with increased mortality . With early diagnosis and initiation of therapy the outcome is favorable.

J Med Microbiol, 2002 Feb, 51(2), 105 - 9
Effect of fluvastatin and pravastatin, HMG-CoA reductase inhibitors, on fluconazole activity against Candida albicans; Nash JD et al.; Synergy between fluvastatin, at clinically unachievable concentrations, and fluconazole against Candida albicans has been reported . The purpose of the present study was to evaluate the in-vitro activity of fluconazole alone and in combination with clinically achievable concentrations of pravastatin and fluvastatin against C . albicans . In-vitro susceptibility and synergy testing were performed against clinical isolates of C . albicans with fluconazole, pravastatin and fluvastatin . Both checkerboard method and time-kill studies were performed . MICs for fluconazole ranged from 0.5 (susceptible) to >256 mg/L (resistant) at 24 h . All isolates had MICs >2 mg/L for both statins . No synergy or antagonism was observed with fluconazole in combination with either agent against any isolate of C . albicans by the checkerboard assay or time-kill studies . Clinically achievable concentrations of pravastatin and fluvastatin did not affect the in-vitro activity of fluconazole against C . albicans.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2002 Feb, 93(2), 155 - 60
Treatment of oral candidiasis with methylene blue-mediated photodynamic therapy in an immunodeficient murine model; Teichert MC et al.; OBJECTIVES:The purpose of this study was to evaluate the efficacy of using methylene blue (MB)-mediated photodynamic therapy to treat oral candidiasis in an immunosuppressed murine model, mimicking what is found in human patients . STUDY DESIGN: Seventy-five experimental mice with severe combined immunodeficiency disease were inoculated orally with Candida albicans by swab 3 times a week for a 4-week period . On treatment day, mice were cultured for baseline fungal growth and received a topical oral cavity administration of 0.05 mL MB solution at one of the following concentrations: 250, 275, 300, 350, 400, 450, or 500 microgram/mL . After 10 minutes the mice were recultured and underwent light activation with 664 nm of diode laser light with a cylindrical diffuser . After photodynamic therapy the mice were cultured again for colony-forming units per milliliter and then killed, their tissue harvested for histopathology . RESULTS AND CONCLUSIONS: The results indicate an MB dose-dependent effect . Concentrations from 250 to 400 microgram/mL reduced fungal growth but did not eliminate Candida albicans . MB concentrations of 450 and 500 microgram/mL totally eradicated Candida albicans from the oral cavity, resulting in reductions from 2.5 log(10) and 2.74 log(10) to 0, respectively . These results suggest that MB-mediated photodynamic therapy can potentially be used to treat oral candidiasis in immunodeficient patients.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2002 Feb, 93(2), 149 - 54
Relationship between salivary flow rates and Candida counts in subjects with xerostomia; Torres SR et al.; OBJECTIVE: This study evaluated the relationship between salivary flow and Candida colony counts in the saliva of patients with xerostomia . STUDY DESIGN: Sialometry and Candida colony-forming unit (CFU) counts were taken from 112 subjects who reported xerostomia in a questionnaire . Chewing-stimulated whole saliva was collected and streaked in Candida plates and counted in 72 hours . Species identification was accomplished under standard methods . RESULTS: There was a significant inverse relationship between salivary flow and Candida CFU counts (P =.007) when subjects with high colony counts were analyzed (cutoff point of 400 or greater CFU/mL) . In addition, the median sialometry of men was significantly greater than that of women (P =.003), even after controlling for confounding variables like underlying disease and medications . Sjogren's syndrome was associated with low salivary flow rate (P =.007) . There was no relationship between the median Candida CFU counts and gender or age . There was a high frequency (28%) of mixed colonization . Candida albicans was the most frequent species, followed by C parapsilosis, C tropicalis, and C krusei . CONCLUSIONS: In subjects with high Candida CFU counts there was an inverse relationship between salivary flow and Candida CFU counts.

Jpn J Infect Dis, 2001 Dec, 54(6), 229 - 36
Characterization of phenotype-based pathogenic determinants of various Candida albicans strains in Jordan; Abu-Elteen KH et al.; Sixty-six clinical isolates of Candida albicans representing 14 different strain types were tested for their phospholipase and proteinase activities in correlation with adherence to buccal epithelial cells (BECs) and lethality to mice . Variations in phospholipase and proteinase production as well as adherence to BECs were observed both among isolates of the same strain type and between isolates of different strain types . All isolates tested, irrespective of strain type, produced low levels of phopholipase (0.5 mm for strain -BCD- and 2.7 mm for strain ABC--) and acid proteinase (0.6 mm for strain A---E and 2.2 mm for strain --C--) . A correlation was noted between adherence, phospholipase and proteinase production, and lethality to mice . C . albicans isolates, which adhered most strongly to BECs, exhibited higher levels of phospholipase and proteinase activities as well as higher pathogenicity . This was most evident in strain type --C--, which exhibited higher adherence ability (mean 717 +/- 21 yeasts/100 BEC), and proteinase activity (mean 2.2 mm), and relatively higher phospholipase activity (mean 2.4 mm) compared with those of other strains . Additionally, this type was more prevalent and showed significantly higher levels of tissue colonization in the liver, kidneys, and spleen compared with most other strain types in both subjects with healthy dentates and complete denture wearers . These results clearly demonstrate the significant role of phospholipase and proteinase activities on the adherence of C . albicans and their overall influence on the pathogenesis of Candida species.

Pediatr Res, 2002 Mar, 51(3), 290 - 5
Cecal colonization and systemic spread of Candida albicans in mice treated with antibiotics and dexamethasone; Bendel CM et al.; Infections with Candida albicans have become a significant problem among very low birth weight infants in the neonatal intensive care unit . Risk factors are multiple and include administration of antibiotics and glucocorticoids, such as dexamethasone . Experiments were designed to study the combined effect of oral broad-spectrum antibiotics and parenteral dexamethasone on cecal colonization and extraintestinal dissemination of C . albicans in separate groups of mice that were orally inoculated with one of four C . albicans strains that were either wild-type INT1/INT1 or had one or more disruptions of the INT1 gene . Intestinal colonization was monitored by quantitative culture of the mouse cecum, and extraintestinal invasion was monitored by quantitative culture of the draining mesenteric lymph nodes and kidneys . At sacrifice, the average numbers of cecal C . albicans differed from 7.7 log(10)/g to 6.7 log(10)/g (p < 0.01) in mice orally inoculated with C . albicans containing two functional copies of INT1 and no functional copies of INT1, respectively . The incidence of extraintestinal dissemination to mesenteric lymph nodes and kidneys correspondingly varied from 57 to 13% (p < 0.01) and 83 to 4% (p < 0.01) in mice inoculated with these two C . albicans strains . Mice orally inoculated with C . albicans containing one functional copy of INT1 had intermediate levels of cecal colonization and extraintestinal dissemination . Thus, cecal colonization and extraintestinal dissemination of C . albicans was facilitated in antibiotic-treated mice given dexamethasone . In addition, the presence of two functional copies of the INT1 gene was associated with the greatest levels of cecal colonization and extraintestinal dissemination of C . albicans.

Med Mycol, 2002 Feb, 40(1), 73 - 81
Transcription initiation of genes associated with azole resistance in Candida albicans; Harry JB et al.; Oral infections with the opportunistic pathogenic yeast Candida albicans are one of the earliest and most frequent infections in immunosuppressed individuals . In these populations, drug-resistant isolates have emerged with the widespread use of antifungal azole drugs . Many molecular mechanisms of resistance have been identified, including overexpression of two types of efflux pumps, the major facilitator MDR1 and the ABC-transporters (CDR1 and CDR2), and the overexpression or mutation of the target enzyme, ERG11 . With overexpression of these four genes implicated in multidrug resistance, identification of regulatory regions of the promoters is important . 5' rapid amplification of cDNA ends (RACE) was used to identify transcription initiation sites for genes associated with multidrug resistance (CDR1, CDR2, MDR1 and ERG11) . These results were confirmed by cloning and sequencing of 5' RACE products and by primer extension . This research will allow further analysis of the regulation of transcription for these genes.

Med Mycol, 2002 Feb, 40(1), 13 - 9
Chemiluminescence of superoxide generated by Candida albicans: differential effects of the superoxide generator paraquat on a wild-type strain and a respiratory mutant; Aoki S et al.; We previously reported that a respiration-competent parent strain (K) of Candida albicans was more susceptible to the intracellular superoxide radical (O2-) generator paraquat (PQ) than was a respiration-deficient mutant (KRD-19), although both showed a similar sensitivity to extracellularly generated O2- . To clarify the cause of the differential PQ lethality, we developed a chemiluminescence method for measuring O2- generated by C . albicans cells by using the probe methyl-Cypridina-luciferin analogue (MCLA), and examined the effects of PQ on O2- generation in both parent and mutant strains . Endogenous O2- generation without stimulation by PQ was unexpectedly low in both strains . PQ-induced O2- generation in the parent strain was maximal in logarithmic phase cells and lowered in stationary phase cells . In contrast, O2- generation in the mutant remained low throughout the growth phase, even when stimulated by PQ . The extent of PQ-induced O2- generation in the parent strain depended on the carbon source added to the assay mixture; in decreasing order, glucose, glycerol, no carbon source . The inhibitor of the cytochrome respiratory chain, antimycin A, suppressed almost completely the PQ-induced O2- generation in the parent strain . It has been established that PQ is converted to its radical form (PQ+) by receiving a single electron in cells . PQ+ then reduces molecular oxygen to O2- by redox cycling . Thus, the high tolerance to PQ of the respiration-deficient mutant can be explained by minimal PQ+/O2- production due to the limited supply of electrons from the impaired respiratory system.

Rev Med Interne, 2002 Jan, 23(1), 30 - 40
{Candida endocarditis: retrospective study in 12 patients}; Abgueguen P et al.; PURPOSE: Candida endocarditis are rare, with a poor prognosis . Actually, the principal problem concerns the growing incidence of nosocomial fungal infections . The objective of the present investigation is to assess a disease which risks becoming more pronounced in the future . METHODS: We have collected observations of Candida sp . endocarditis between 1985 to 1997 from three French university hospitals . RESULTS: Twelve of the observations fit the Duke criteria of acute endocarditis . Patients were eight men and four women, with a mean age of 46 years . An immunodepression was found in seven cases, and four patients were active drug addicts . Six had an underlying heart disease at risk to acute endocarditis . Candidemia risk factors were found in nine cases, with an average of 2.7 risk factors per patient . The fungal agents detected were Candida albicans (eight cases), C . tropicalis (one case), C . parapsilosis (two cases), and C . glabrata (one case) . These vegetations were on aortic (seven cases), mitral (three cases), tricuspid valves (two cases) or in other areas (three cases), with multiple localizations (two cases) . In three observations, vegetations were associated with myocardium abscesses . Eight patients had embolic complications, two had a cardiac insufficiency leading to death . The treatment was medical in all of the cases and combined with a surgical treatment in ten cases . The surgery was performed, on an average, 17 days after diagnosis, allowing seven surviving patients . Among them, five received a secondary prophylaxis and no recurrence was recorded . CONCLUSIONS: Prognosis remains severe because of the voluminous, friable and necrotic vegetations, which favor embolic migrations and which are not easily accessible to antifungals, which penetrate poorly into these vegetations . Therapy is based on a medical treatment combined with a valve replacement which needs to be done early on, and is followed by a relapse prevention which can occur several years after the initial episode.

Bioorg Med Chem Lett, 2002 Mar 11, 12(5), 803 - 6
Synthesis of Sordaricin analogues as potent antifungal agents against Candida albicans; Kaneko S et al.; Sordaricin derivatives possessing a cyclohexane ring appendage attached via an ether, thioether, amine, oxime, ester or amide linkage were synthesized and their antifungal activity was evaluated in vitro . Compounds containing a thioether bond or an oxime bond as a linkage exhibited potent MICs (< or = 0.125 microg/mL) against four Candida albicans strains including azole-low-susceptible strains . They were also active (MIC < or = 0.125 microg/mL) against Candida glabrata . Their in vivo efficacy was confirmed in a murine intravenous infection model with Candida albicans.

Mycoses, 2002 Feb, 45(1-2), 41 - 4
Case Report . Multiple etiology post-surgery endophthalmitis; Vescia N et al.; The case describes a septic endophthalmitis arisen in a convalescence period following surgery of cataract extraction . The infection was due to Staphylococcus aureus and three fungal components, Candida albicans, Candida glabrata and Acremonium kiliense, which were subsequently isolated . A careful and prompt laboratory investigation allowed the clinicians to adjust the antimycotic therapy and attain an excellent clinical result.

Mol Cell Biochem, 2001 Dec, 228(1-2), 111 - 7
Antifungal activities of origanum oil against Candida albicans; Manohar V et al.; The antimicrobial properties of volatile aromatic oils from medicinal as well as other edible plants has been recognized since antiquity . Origanum oil, which is used as a food flavoring agent, possesses a broad spectrum of in vitro antimicrobial activities attributed to the high content of phenolic derivatives such as carvacrol and thymol . In the present study, antifungal properties of origanum oil were examined both in vitro and in vivo . Using Candida albicans in broth cultures and a micro dilution method, comparative efficacy of origanum oil, carvacrol, nystatin and amphotericin B were examined in vitro . Origanum oil at 0.25 mg/ml was found to completely inhibit the growth of C . albicans in culture . Growth inhibitions of 75% and >50% were observed at 0.125 mg/ml and 0.0625 mg/ml level, respectively . In addition, both the germination and the mycelial growth of C . albicans were found to be inhibited by origanum oil and carvacrol in a dose-dependent manner . Furthermore, the therapeutic efficacy of origanum oil was examined in an experimental murine systemic candidiasis model . Groups of mice (n = 6) infected with C . albicans (5 x LD50) were fed varying amounts of origanum oil in a final vol . of 0.1 ml of olive oil (vehicle) . The daily administration of 8.6 mg of origanum oil in 100 microl of olive oil/kg body weight for 30 days resulted in 80% survivability, with no renal burden of C . albicans as opposed to the group of mice fed olive oil alone, who died within 10 days . Similar results were obtained with carvacrol . However, mice fed origanum oil exhibited cosmetically better clinical appearance compared to those cured with carvacrol . The results from our study encourage examination of the efficacy of origanum oil in other forms of systemic and superficial fungal infections and exploration of its broad spectrum effect against other pathogenic manifestations including malignancy.

Infect Immun, 2002 Mar, 70(3), 1558 - 65
Temporal expression of the Candida albicans genes CHK1 and CSSK1, adherence, and morphogenesis in a model of reconstituted human esophageal epithelial candidiasis; Li D et al.; We previously demonstrated that genes encoding a putative two-component histidine kinase (CHK1) or a response regulator (CSSK1) are each required for virulence in a murine model of hematogenously disseminated candidiasis and that strains with each gene deleted are also defective in morphogenesis under certain growth conditions . In the present study, the role of these two genes in the adherence to and colonization of reconstituted human esophageal tissue (RHE) is described . We compared strains of Candida albicans with deletions of chk1 (strain CHK21) and cssk1 (strain CSSK21) to wild-type cells (CAF2), as well as strains with CHK1 and CSSK1 reconstituted (strains CHK23 and CSSK23, respectively) . Adherence and colonization of RHE were evaluated in periodic acid-Schiff-stained sections, as well as by SEM . We observed that both deletion-containing strains colonized the RHE to a lesser extent than did CAF2 and that the percent germination by both strains was reduced in comparison to that of control strains at 1 h postinfection . Expression of CHK1 or CSSK1 was quantitated by reverse transcription (RT)-PCR from RHE tissues infected with wild-type C . albicans yeast cells . Expression of both CHK1 and CSSK1 increased over the 48-h period following infection of the tissue, although expression of CHK1 was greater than that of CSSK1 . By RT-PCR, we have also shown that expression of CHK1 and CSSK1 in the strains with cssk1 and chk1 deleted, respectively, was similar to that of CAF2, indicating that CHK1 and CSSK1 do not regulate each other but probably encode signal proteins of different pathways . Our observations indicate that CHK1 and CSSK1 are each partially required for colonization and conversion to filamentous growth on RHE tissue.

Biochim Biophys Acta, 2002 Jan 30, 1542(1-3), 73 - 81
Interactions between regulatory and catalytic subunits of the Candida albicans cAMP-dependent protein kinase are modulated by autophosphorylation of the regulatory subunit; Zelada A et al.; The cAMP-dependent protein kinase (PKA) from Candida albicans is a tetramer composed of two catalytic subunits (C) and two type II regulatory subunits (R) . To evaluate the role of a putative autophosphorylation site of the R subunit (Ser(180)) in the interaction with C, this site was mutated to an Ala residue . Recombinant wild-type and mutant forms of the R subunit were expressed in Escherichia coli and purified . The wild-type recombinant R subunit was fully phosphorylated by the purified C subunit, while the mutant form was not, confirming that Ser(180) is the target for the autophosphorylation reaction . Association and dissociation experiments conducted with both recombinant R subunits and purified C subunit showed that intramolecular phosphorylation of the R subunit led to a decreased affinity for C . This diminished affinity was reflected by an 8-fold increase in the concentration of R subunit needed to reach half-maximal inhibition of the kinase activity and in a 5-fold decrease in the cAMP concentration necessary to obtain half-maximal dissociation of the reconstituted holoenzyme . Dissociation of the mutant holoenzyme by cAMP was not affected by the presence of MgATP . Metabolic labeling of yeast cells with {(32)P}orthophosphate indicated that the R subunit exists as a serine phosphorylated protein . The possible involvement of R subunit autophosphorylation in modulating C . albicans PKA activity in vivo is discussed.

Antimicrob Agents Chemother, 2002 Mar, 46(3), 797 - 807
Antileishmanial activities of several classes of aromatic dications; Brendle JJ et al.; Aromatic dicationic molecules possess impressive activity against a broad spectrum of microbial pathogens, including Pneumocystis carinii, Cryptosporidium parvum, and Candida albicans . In this work, 58 aromatic cations were examined for inhibitory activity against axenic amastigote-like Leishmania donovani parasites . In general, the most potent of the compounds were substituted diphenyl furan and thiophene dications . 2,5-Bis-(4-amidinophenyl)thiophene was the most active compound . This agent displayed a 50% inhibitory concentration (IC50) of 0.42 +/- 0.08 microM against L . donovani and an in vitro antileishmanial potency 6.2-fold greater than that of the clinical antileishmanial dication pentamidine and was 155-fold more toxic to the parasites than to a mouse macrophage cell line . 2,4-Bis-(4-amidinopheny)furan was twice as active as pentamidine (IC50), 1.30 +/- 0.21 microM), while 2,5-bis-(4-amidinopheny)furan and pentamidine were essentially equipotent in our in vitro antileishmanial assay . Carbazoles, dibenzofurans, dibenzothiophenes, and benzimidazoles containing amidine or substituted amidine groups were generally less active than the diphenyl furans and thiophenes . In all cases, aromatic dications possessing strong antileishmanial activity were severalfold more toxic to the parasites than to a cultured mouse macrophage cell line . These structure-activity relationships demonstrate the potent antileishmanial activity of several aromatic dications and provide valuable information for the future design and synthesis of more potent antiparasitic agents.

Neuroimmunomodulation, 2001, 9(4), 193 - 202
Impaired activity of phagocytic cells in Candida albicans infection after exposure to chronic varied stress; Rodriguez-Galan MC et al.; OBJECTIVE: Candidiasis is a prototypic opportunistic fungal disease that may follow severe modulations of the immune system of the host . The purpose of this study was to evaluate which innate immune mechanisms involved in the protection against fungal invasion are impaired under stress conditions . METHODS: Wistar rats were infected intraperitoneally with Candida albicans and immediately exposed to chronic varied stress (CVS) over 10 days (CVS; Ca-S); the fungal burden (CFU), histopathological lesion and ACTH levels were evaluated . Additionally, functional assessment of peritoneal cells (PC) included the phagocytic and anticandidacidal activities and the production of H(2)O(2) and NO . RESULTS: In the only infected animals (Ca), C . albicans colonization stimulated an efficient inflammatory response, while in Ca-S rats poor tissue reactions were associated with increased CFU in livers and kidneys (p < 0.05, Ca vs . Ca-S) . Whereas the phagocytic process was not modified, the candidacidal activity of PC was significantly decreased after the application of CVS (p < 0.001, Ca vs . Ca-S) . The H(2)O(2) production by macrophages and neutrophils was downregulated by the infection, and while at early intervals these cells possessed a residual oxidative capacity, by day 10, the production of this metabolite was blocked . Spontaneous NO production by macrophages was significantly increased in both Ca and Ca-S animals (p < 0.001), but in stressed rats, this reactive nitrogen intermediate was noticeably downregulated (p < 0.05, Ca vs . Ca-S) . The hyperactivity of hypothalamus-pituitary-adrenal axis after exposure to stress was confirmed by an increase in baseline plasma ACTH levels . CONCLUSION: These results show that during infection with C . albicans, the exposure to CVS contributes to the spread of the fungus and downregulates critical functions of phagocytic cells involved in the control of this opportunistic pathogen .

Hong Kong Med J, 1997 Sep, 3(3), 312 - 314
Candida krusei infections and fluconazole therapy; Samaranayake LP; Candida species are by far the most common agents of mucosal fungal infection in man . While Candida albicans is the most notorious pathogen in this group, non-albicans species such as Candida krusei are gradually emerging as pathogens of concern, especially in compromised hosts . It is thought that the wide use of the newer triazole drug, fluconazole, in HIV-infected individuals is contributing to this phenomenon . Studies in both humans and animals have now demonstrated prophylactic and therapeutic failure of fluconazole against C . krusei due to increasing resistance of the organism to this azole . Thus, the indiscriminate use of fluconazole, a drug with relatively minimal toxicity and excellent pharmacokinetics, may lead to the development of widespread resistance to this azole among Candida species.

EMBO J, 2002 Feb 15, 21(4), 546 - 59
Calcineurin is essential for survival during membrane stress in Candida albicans; Cruz MC et al.; The immunosuppressants cyclosporin A (CsA) and FK506 inhibit the protein phosphatase calcineurin and block T-cell activation and transplant rejection . Calcineurin is conserved in microorganisms and plays a general role in stress survival . CsA and FK506 are toxic to several fungi, but the common human fungal pathogen Candida albicans is resistant . However, combination of either CsA or FK506 with the antifungal drug fluconazole that perturbs synthesis of the membrane lipid ergosterol results in potent, synergistic fungicidal activity . Here we show that the C.albicans FK506 binding protein FKBP12 homolog is required for FK506 synergistic action with fluconazole . A mutation in the calcineurin B regulatory subunit that confers dominant FK506 resistance (CNB1-1/CNB1) abolished FK506-fluconazole synergism . Candida albicans mutants lacking calcineurin B (cnb1/cnb1) were found to be viable and markedly hypersensitive to fluconazole or membrane perturbation with SDS . FK506 was synergistic with fluconazole against azole-resistant C.albicans mutants, against other Candida species, or when combined with different azoles . We propose that calcineurin is part of a membrane stress survival pathway that could be targeted for therapy.

Clin Immunol, 2002 Feb, 102(2), 154 - 61
CD4+ T-lymphocyte nadir and the effect of highly active antiretroviral therapy on phenotypic and functional immune restoration in HIV-1 infection; Lange CG et al.; To evaluate the effects of the timing of highly active antiretroviral therapy (HAART) on immune reconstitution, we compared lymphocyte subpopulations and lymphocyte proliferation (LP) in response to Candida albicans, cytomegalovirus, HIV p24, Mycobacterium avium complex, pokeweed mitogen, streptokinase, and tetanus toxoid in 43 patients with pretherapy advanced, moderately advanced, and early chronic HIV-1 infection . All patients had recent CD4+ T-cell counts >450/microl and HIV RNA <400 copies/ml for >12 months . CD4+ nadirs were positively correlated with recent numbers of CD4+ T-cells (P < 0.001), memory cells (P < 0.001), and naive CD4+ T-cells (P < 0.05) and CD4+ CD28+ T-lymphocytes (P < 0.05) and were negatively correlated with recent CD8+ T-lymphocyte counts (P < 0.05) . Only CD4+ naive T-cells normalized when HAART was initiated at lower CD4+ T-cell levels . Fifty-three percent of patients had LP responses to HIV p24 antigen . While LP responses to prevalent antigens were usually present, responses to tetanus toxoid were more common with higher CD4+ T-lymphocyte nadirs (P < 0.05) . Delaying HAART may limit phenotypic and functional immune restoration in HIV-1 infection .

Bioorg Med Chem Lett, 2002 Feb 25, 12(4), 629 - 32
A new class of type I protein geranylgeranyltransferase (GGTase I) inhibitor; Sunami S et al.; Replacement of the thiol groups in 1, a potent and highly selective Candida albicans GGTase I inhibitor discovered through screening, with an imidazole ring was achieved by using solid phase synthesis . A non-thiol compound, 7, was found as a representative of a new class of potent C . albicans GGTase I inhibitor with high selectivity against human GGTase I.

Planta Med, 2002 Jan, 68(1), 49 - 54
Natural products inhibiting Candida albicans secreted aspartic proteases from Tovomita krukovii; Zhang Z et al.; Assay-guided fractionation of the ethanol extract of Tovomita krukovii resulted in the identification of four new xanthones (1 - 4) and ten known compounds (5 - 14) . The structures of compounds 1 - 14 were determined by spectral data to be 3,5-dihydroxy-4-methoxyxanthone (1), 1,3,5,7-tetrahydroxy-8-isoprenylxanthone (2), 1,3,5-trihydroxy-8-isoprenylxanthone (3), 1,5,7-trihydroxy-8-isoprenylxanthone (4), 1,3,7-trihydroxy-2-isoprenylxanthone (5), 1,5-dihydroxyxanthone (6), 1,6-dihydroxy-5-methoxyxanthone (7), 1,3,5-trihydroxyxanthone (8), 1,3,6-trihydroxy-5-methoxyxanthone (9), 1,6-dihydroxy-3,5-dimethoxyxanthone (10), 1,3,7-trihydroxyxanthone (11), 3-geranyl-2,4,6-trihydroxybenzophenone (12), betulinic acid (13), and 3,4-dihydroxybenzoic acid (14) . Compounds 2, 3, 12 and 13 showed inhibitory effects against Candida albicans secreted aspartic proteases (SAP) with IC50 values of 15 microg/ml, 25 microg/ml, 40 microg/ml, and 6.5 microg/ml, respectively, while the other compounds were inactive . In addition, compound 12 showed activity against C . albicans, C . neoformans, S . aureus and methicillin resistant S . aureus (MRS).

Planta Med, 2002 Jan, 68(1), 41 - 4
New and antifungal xanthones from Calophyllum caledonicum; Morel C et al.; Two new xanthones, namely caledonixanthones E (1) and F (2), were isolated from the stem bark of Calophyllum caledonicum (Guttiferae) . The structural elucidation of these compounds was mainly established on the basis of 1D, 2D NMR and HRMS spectroscopic analysis . Among the isolated compounds, eight other known xanthones were also identified in the course of this phytochemical study . In addition to this report, a preliminary evaluation of the antifungal properties of these polyphenolic compounds against Aspergillus fumigatus and Candida albicans is presented.

Biochemistry, 2002 Feb 19, 41(7), 2254 - 63
Conjugation of a magainin analogue with lipophilic acids controls hydrophobicity, solution assembly, and cell selectivity; Avrahami D et al.; Our basic understanding of how to combat fungal infections has not kept pace with the recent sharp rise in life-threatening cases found particularly among immuno-compromised individuals . Current investigations for new potential antifungal agents have focused on antimicrobial peptides, which are used as a cell-free defense mechanism in all organisms . Unfortunately, despite their high antibacterial activity, most of them are not active toward fungi, the reason of which is not clear . Here, we present a new approach to modify an antibacterial peptide, a magainin analogue, to display antifungal activity by its conjugation with lipophilic acids . This approach has the advantage of producing well-defined changes in hydrophobicity, secondary structure, and self-association . These modifications were characterized in solution at physiological concentrations using CD spectroscopy, tryptophan fluorescence, and analytical ultracentrifugation . In order of increasing hydrophobicity, the attachment to the magainin-2 analogue of (i) heptanoic acid results in a monomeric, unordered structure, (ii) undecanoic acid yields concentration-dependent oligomers of alpha helices, and (iii) palmitic acid yields concentration-independent alpha-helical monomers, a novel lipopeptide structure, which is resistant to proteolytic digestion . Membrane-lipopeptide interactions and the membrane-bound structures were studied using fluorescence and ATR-FTIR in PC/PE/PI/ergosterol (5/2.5/2.5/1, w/w) SUV, which constitute the major components of Candida albicans bilayers . A direct correlation was found between oligomerization of the lipopeptides in solution and potent antifungal activity . These results provide insight to a new approach of modulating hydrophobicity and self-assembly of antimicrobial peptides in solution, without altering the sequence of the peptidic chain . These studies also provide a general means of developing a new group of lipopeptide candidates as therapeutic agents against fungal infections.

Arch Oral Biol, 2002 Mar, 47(3), 189 - 96
Genotyping and antifungal susceptibility of human subgingival Candida albicans isolates; Pizzo G et al.; Subgingival colonization by Candida albicans has been described in human immunodeficiency virus (HIV)-infected individuals, but subgingival isolates have scarcely been characterized, particularly with respect to genotype and antifungal susceptibility . A series of 29 subgingival strains of C . albicans isolated from nine HIV-infected individuals was typed by electrophoretic karyotyping and tested for susceptibility to fluconazole, itraconazole, the new investigational triazole posaconazole and amphotericin B . DNA typing showed genetic heterogeneity within subgingival isolates, as almost every individual harbored his/her own specific isolate . Genetic identity was usually demonstrated within oral and subgingival isolates simultaneously collected from the same individual, but a number of DNA types were found to be unique to subgingival strains . These findings suggest that colonization is not just the result of Candida spreading from oral surfaces, and that subgingivally adapted strains could be involved . All isolates were susceptible to all the triazole drugs tested and amphotericin B . Additional studies on subgingival Candida colonization and further characterization of subgingival isolates are now required to clarify the role of Candida as opportunistic periodontal pathogen.

Drug Deliv, 2002 Jan-Mar, 9(1), 31 - 8
Design and fungicidal activity of mucoadhesive lactoferrin tablets for the treatment of oropharyngeal candidosis; Kuipers ME et al.; Lactoferrin (Lf) is a potential drug candidate for the treatment of oropharyngeal Candida infections . However, for an effective therapeutic treatment an appropriate dosage form is required . Therefore a mucoadhesive tablet for buccal application was developed . Tablets of sufficient strength could be produced on high speed tabletting machines, but they could only be obtained when the protein contained at least 7% moisture . The tablet contained sodium alginate both for its release-controlling properties as well as for its mucoadhesive properties . Furthermore, phosphate buffer was added to keep the pH of the saliva in the mouth within the range of 6.5 to 7.5 . In this pH range, Lf has shown to have its highest activity against Candida growth inhibition . The tablet formulation containing Lf had the same antifungal properties as compared with Lf alone, because in most cases identical inhibitory concentrations were observed against several clinical isolates of Candida albicans and Candida glabrata . In human volunteers the tablets, containing 250 mg Lf and placed in each pouch, were able to keep the Lf concentration in the saliva at effective levels for at least 2 hr, while the pH of the saliva remained within the desired range . We concluded that the developed mucoadhesive tablet can improve the therapeutic efficacy of Lf and that it is suitable for further clinical research.

Bioorg Med Chem, 2002 Apr, 10(4), 1153 - 70
Insights into the selective inhibition of Candida albicans secreted aspartyl protease: a docking analysis study; Pranav Kumar SK et al.; Severe fungal infections have taken precedence over other bacterial infections . Of the several fungal species, Candida albicans and others belonging to the genus Candida are responsible for several clinically important fungal infections . Emerging cases of drug resistance to the currently available drugs has limited the spectrum of currently available antifungal agents . Thus, it is imperative that new biochemical targets are identified so that better effective and selective agents can be developed . Many enzymes contribute towards the complex disease process of fungal infections; the secreted aspartyl protease (SAP), expressed both in vitro and during infection, has been implicated as one of the major virulence factors of C . albicans . Three-dimensional crystal structures of C . albicans SAP and closely related clinical isolate designated as SAP2X complexed with the same potent inhibitor A-70450 have been reported . Several analogues of A-70450 with potent C . albicans SAP2X inhibitory activity are also known . However, the structural effects of the binding of these compounds with the enzyme active site are not completely understood . Our efforts in this direction involve the docking analysis of C . albicans SAP2X inhibitors complexed with SAP2X enzyme, which is reported in this work . Docking analysis was performed on a set of molecules with differing selectivities and inhibitory potencies towards C . albicans, renin and cathepsin D . The structural effects of ligand binding were analyzed on the basis of hydrophobic and hydrogen bond interactions, binding energy analysis, interaction energies, rms deviations, etc . in the resulting energy-minimized structures of the receptor-ligand complexes . Structural analysis of the resulting models indicates that hydrophobic and hydrogen bonding interactions together with binding and interaction energies are responsible for selective inhibition of C . albicans SAP2X . Hydrophobic and hydrogen bonding interactions in the various subsites of the enzyme, contributing to both increase as well as decrease in selectivity of the molecules have been detailed . Hydrogen bonding interaction plays an important role for amino acid residues such as Gly-85, Asp-86, Asp-32, Asp-218, Tyr-225, Ala-133, and so on . Significant hydrophobic interactions with the S3, S2 and S2' subsites contribute to selectivity of the compounds . These molecular modeling analyses should, in our view, contribute for further development of selective C . albicans secreted aspartyl protease inhibitors.

J Endod, 2002 Feb, 28(2), 68 - 71
Effectiveness of intracanal irrigants and medications against the yeast Candida albicans; Ferguson JW et al.; An in vitro study was conducted to determine the susceptibility of the yeast Candida albicans to various intracanal irrigants and medications . The minimum inhibitory concentration (MIC) of sodium hypochlorite, hydrogen peroxide, chlorhexidine digluconate, and aqueous calcium hydroxide that is required to kill a standardized inoculum of C . albicans was determined . Growth of the yeast was measured by optical density . Sodium hypochlorite, hydrogen peroxide, and chlorhexidine digluconate were effective anticandidal agents with MICs of <10 microgram/ml, 234 microgram/ml, and <0.63 microgram/ml, respectively . Aqueous calcium hydroxide had no activity . A standardized inoculum of C . albicans cells was also placed in direct contact with either calcium hydroxide paste or camphorated para-monochlorophenol (CPMC), and candidal growth was assessed by colony counts on Sabouraud's dextrose agar . Calcium hydroxide paste and CPMC, when maintained in direct contact with C . albicans, were effective antifungal agents.

Rev Argent Microbiol, 2001 Oct-Dec, 33(4), 217 - 22
{Vaginal candidiasis: etiology and sensitivity profile to antifungal agents in clinical use}; Saporiti AM et al.; Candida vaginitis is one of the most frequent infection of the female genital tract with a high incidence . Approximately 75% of sexually active women suffer at least one episode of Candida vaginitis and 10% of them have recurrent episodes . Pregnancy, diabetes mellitus and antibiotic treatment are the most common predisposing factors, C . albicans is the etiologic agent most frequently found . The widespread reports of fluconazole resistance in Candida species and the selection of non Candida albicans prompted the study of species distribution of vulvovaginal candidiasis and their in vitro susceptibility against current antifungal agents . A total of 314 women with vaginal infection were studied . Yeasts were isolated from 104 patients with vulvovaginal candidiasis . The following species were identified: C . albicans 87.5%, C . glabrata 8.6% and 3.9% included C . krusei, C . famata, C . tropicalis and S . cerevisiae . The minimal inhibitory concentration (MIC) was determined for nystatin, isoconazole, fluconazole and ketoconazole, using a broth microdilution method based on NCCLS procedure . Although most of the isolates were C . albicans, the high percentage of C . glabrata recovered suggests the need to identify the yeasts isolated . Fluconazole resistant C . albicans were isolated in 13.46% of the cases . Thus, further studies are required to correlate the possible role of these strains in recurrent vulvovaginal candidiasis.

Rev Argent Microbiol, 2001 Oct-Dec, 33(4), 209 - 16
{Comparative study of the Candida albicans genotypes isolated from immunocompromised patients and health carriers}; Carnovale S et al.; The aim of this study is to compare Candida albicans strain genotype isolates from oral cavity of immunocompromised patients due to different immunologic impairments with apparently normal carriers . Four populations were studied: 1) HIV positive hospitalized patients, 2) HIV negative immunocompromised patients (leukemia, lymphoma, organ transplant recipients), 3) drug addicts prior to AIDS pandemia in Argentina, 4) apparently normal carriers . DNA extracted was digested with the enzyme Eco RI, electrophoresed, transferred to nitrocellulose membrane and hybridized with the 27A probe labelled with 32P . The comparison between the profiles obtained permitted the differentiation of 16 genotypes . The distribution of the strains led to the conclusion that: a) all the isolated strains from AIDS patients were closely related and distributed in only three genotypes (1, 3, 11); b) a major genetic relationship between the isolates from AIDS patients and HIV negative immunocompromised patients was observed; c) strains from carriers showed a minor genetic similarity with those obtained from AIDS patients; d) characteristic profiles belonging to any of the studied groups were not found; e) significant genomic changes have not been observed during the last twenty years.

Microbiology, 2002 Feb, 148(Pt 2), 497 - 506
In vitro reconstructed human epithelia reveal contributions of Candida albicans EFG1 and CPH1 to adhesion and invasion; Dieterich C et al.; The individual and synergistic contributions of two transcription factors, EFG1 and CPH1, have been characterized with regard to adhesion to, and invasion of, human epithelia by Candida albicans . For this purpose two in vitro reconstructed tissue models were developed . A multi-layered model of human epidermis was used to simulate superficial infections of the skin, whereas a reconstructed human intestinal model was used to mimic the first steps of systemic infections . It was shown that C . albicans deleted for both transcription factors CPH1 and EFG1, in contrast to the congenic clinical isolate Sc5314, was neither able to adhere to, nor to penetrate, either of the model systems . A strain deleted for EFG1 alone showed significant reduction in adhesion and was not able to penetrate through the stratum corneum . However, strains deleted for CPH1 showed phenotypes paralleling the phenotypes of the clinical isolate Sc5314 . Using different types of multi-layered human tissues reconstructed in vitro the individual contributions of Efg1p and Cph1p to two important virulence factors of C . albicans, namely adhesion and invasion, could be defined.

Inflammation, 2001 Dec, 25(6), 381 - 7
Contribution of myeloperoxidase to coronary artery vasculitis associated with MPO-ANCA production; Ishida-Okawara A et al.; The role of myeloperoxidase (MPO) in the pathogenesis of vasculitis associated with MPO-specific anti-neutrophil cytoplasmic autoantibody (MPO-ANCA) was examined in a murine animal model . Coronary artery vasculitis was induced in C57BL/6 mice with and without endogenous MPO by intraperitoneal injection of Candida albicans-derived substances (CADS) . The corresponding levels of MPO-ANCA in sera of mice with and without vasculitis were measured and compared in both wild-type and MPO-deficient animals . The MPO-ANCA titers in sera were significantly higher in mice with vasculitis than in vasculitis-negative mice, indicating that MPO-ANCA correlated with vasculitis formation . However, the increase of MPO-ANCA titers observed in sera of wild C57BL/6 mice were strongly suppressed in MPO-deficient C57BL/6 mice, accompanied with prevention of vasculitis formation . These results show that MPO acted as an antigen for MPO-ANCA production by CADS and was followed by the vasculitis formation . Vasculitis did develop in a few MPO-deficient mice, though the incidence of vasculitis was much lower in MPO-deficient mice than in C57BL/6 mice.

Expert Opin Investig Drugs, 2002 Feb, 11(2), 309 - 18
Antimicrobial peptides: therapeutic potential for the treatment of Candida infections; Lupetti A et al.; The increasing frequency of fungal infections in immunocompromised patients together with the emergence of strains resistant to currently used antifungal drugs point to an increased need for a new class of antimycotics . Antimicrobial peptides are promising candidates for the treatment of fungal infections since they have both mechanisms of action distinct from available antifungal agents and the ability to regulate the host immune defence systems as well . This review focuses on Candida albicans as a large amount of work on the mechanisms of action of classical antifungals as well as antimicrobial peptides, such as defensins, protegrins, histatins and lactoferrin (LF)-derived peptides, has been performed in this yeast . Analogues of these antimicrobial peptides and combinations of antimicrobial peptides with classical antimycotics are under investigation for treatment of candidiasis.

Nihon Kokyuki Gakkai Zasshi, 2001 Oct, 39(10), 726 - 31
{Role of neutrophils in acute lung injury induced by Candida sepsis}; Ueda M et al.; To evaluate the role of inflammatory cells in acute lung injury induced by Candida sepsis, we investigated the time course of cell counts in bronchoalveolar lavage (BAL) fluid, the lung wet-to-dry weight (W/D) ratio and the CFU of Candida after a Candida infection . Male BALB/c mice were injected intravenously with 107 CFU of Candida albicans . They were sacrificed by exsanguination 0, 6, 12, 18, and 24 h after infection . The lung W/D ratio was determined in order to assess the severity of pulmonary edema . BAL was performed 3 times with 0.5 ml of sterile saline . The lung W/D ratio increased significantly 24 h after the injection of Candida, and the neutrophil counts in the BAL fluid increased significantly 12, 18, and 24 h after Candida injection . To assess the effect of neutrophils on Candida-induced acute lung injury, we investigated that of neutrocytosis and neutropenia on cell counts in BAL fluid, of the W/D ratio, and of the CFU of Candida in the lung 24 h after infection . We created neutrocytotic mice using G-CSF, and neutropenic mice with cyclophosphamide . Peripheral neutrophil counts decreased significantly in cyclophosphamide-treated mice, and increased significantly in G-CSF-treated mice . The lung W/D ratio increased significantly in cyclophosphamide-treated mice . Neutrophil counts in BAL fluid increased significantly in G-CSF-treated mice, and decreased significantly in cyclophosphamide-treated mice . The CFU of Candida in the lung decreased significantly in G-CSF-treated mice, and increased significantly in cyclophosphamide-treated mice . We concluded that neutrophils play a protective role in Candida albicans-induced acute lung injury.

Eur J Cancer Care (Engl), 2001 Mar, 10(1), 56 - 62
Assessing risk factors for systemic fungal infections; Maertens J et al.; The incidence of invasive fungal infection has increased in recent years . Most infections are caused by Candida albicans and Aspergillus spp . but the emergence of other fungal infections is changing the spectrum of disease . Immunosuppression and breakdown of anatomical barriers such as the skin are the major risk factors for fungal infections . Health care workers encounter at-risk patients in various settings, including AIDS clinics and intensive care, transplantation and oncology units . Patients with prolonged and deep neutropenia (haematological malignancy patients) are most at risk and are therefore most likely to receive prophylactic therapy . Practical measures can be taken to avoid exposing the patient to fungi (air filtration, regular hand washing, avoiding plants and flowers) and antifungal agents can be administered to prevent systemic fungal infection . Most fungal infections have non-specific symptoms; this makes recognition of the signs and symptoms of the disease important but also makes diagnosis difficult and empirical treatment necessary . Some antifungal agents have limitations but new formulations will improve therapy and play a key role in future antifungal strategies.

J Antibiot (Tokyo), 2001 Nov, 54(11), 904 - 10
CJ-15,183, a new inhibitor of squalene synthase produced by a fungus, Aspergillus aculeatus; Watanabe S et al.; A new squalene synthase (SSase) inhibitor, CJ-15,183 (I) was isolated from the fermentation broth of a fungus, Aspergillus aculeatus CL38916 . The compound potently inhibited rat liver and Candida albicans microsomal SSases and also inhibited the human enzyme . It also showed antifungal activities against filamentous fungi and a yeast . The structure was determined to be an aliphatic tetracarboxylic acid compound consisting of an alkyl gamma-lactone, malic acid and isocitric acid moieties by spectroscopic studies.

J Antibiot (Tokyo), 2001 Nov, 54(11), 890 - 5
Bisabosquals, novel squalene synthase inhibitors . I . Taxonomy, fermentation, isolation and biological activities; Minagawa K et al.; In the course of screening for yeast squalene synthase inhibitors, bisabosqual A was isolated from the culture broth of Stachybotrys sp . RF-7260 . The related compounds bisabosquals B, C and D were also isolated from Stachybotrys ruwenzoriensis RF-6853 . Bisabosquals inhibited squalene synthases . IC50 values of bisabosqual A against the microsomal squalene synthases from Saccharomyces cerevisiae, Candida albicans, HepG2 cell and rat liver were 0.43, 0.25, 0.95 and 2.5 microg/ml, respectively . Bisabosqual C exhibited inhibitory activities similar to bisabosqual A . Bisabosqual A showed broad spectrum antifungal activity in vitro.

Rev Cubana Med Trop, 2000 Sep-Dec, 52(3), 191 - 6
{Sensitivity of clinical isolates of Candida albicans to 5-fluorocytosine}; Fernandez Andreu CM et al.; 5-fluorocytosine has been one of the most used antimycotics in the therapeutics of mycosis, however the appearance of resistant strains may be an important limitant for its use . That's why it is necessary to know the sensitivity to this drug . To this end, it was determined the minimum inhibitory concentration (MIC) of 5-fluorocytosine against 50 clinical isolates of Candida albicans by a micromethod of dilution in RPMI 1640 media, comparing 2 reading methods . The values of the geometrical mean obtained were 0.95 mg/mL for the visual reading and 0.75 mg/mL for the automated reading . There were no significant differences between them (p < 0.05) . Pearson's lineal correlation coefficient was r = 0.81 . It indicates that although there was a good correlation, there was also discrepancy between both readings . The IMC50 was of 0.5 microgram/mL and the IMC90 of 4 mg/mL . 90 and 92% of sensitive strains, between 6 and 8% of resistant strains and 2% of strains with intermediate sensitivity were found depending on the reading method used . These values show a low resistance against this drug, which may have a direct repercussion on the therapeutic behaviour to be followed in candidiasis.

Rev Cubana Med Trop, 2000 Sep-Dec, 52(3), 186 - 90
{Obtention, characterization of antigens and evaluation of the immunotransfer technique for the detection of systemic candidiasis}; Alvarez E et al.; A culture media containing peptone, glucose, yeast extract and serum was evaluated to promote the development of the mycelium formation in Candida albicans and it was compared with the commercial media Tc 199 . No significant difference was found between them . Somatic antigens characterized by PAGE-SDS were obtained from yeast and mycelial forms . Proteins reported as having high diagnostic value were expressed in both antigens . It was determined by immunotransference that the antigenic proteins predominating in the mycelium formation had molecular weights of 19, 21, 27 and 57 Kda . The immunotransference was standardized for detecting antibodies in patients with systemic candidiasis . 24 positive sera were evaluated by this technique and it was compared with the ELISA technique, which proved to be more sensitive and less specific . The most recognized proteins were those of 136 and 21 Kda and the complex from 42 to 45 Kda.

J Clin Microbiol, 2002 Feb, 40(2), 341 - 50
Carriage frequency, intensity of carriage, and strains of oral yeast species vary in the progression to oral candidiasis in human immunodeficiency virus-positive individuals; Vargas KG et al.; Candida samples were taken over a period of 2 years from 54 human immunodeficiency virus (HIV)-positive asymptomatic subjects to evaluate changes in yeast carriage, intensity of carriage, and genotype over time . Overall, we found that HIV-positive patients with CD4(+)-cell counts of between 200 and 400/microl had significantly more yeast colonization than healthy control subjects . Of the 54 patients, 11 developed thrush . We found that intensity of carriage in these 11 patients increased significantly in the progression from asymptomatic yeast carrier to an episode of oral thrush . Also, the most common yeast species isolated was Candida albicans; however, we did see a number of patients harboring multiple species at the same time . Using the C . albicans-specific probe Ca3, we found that 54% (n = 6) of the 11 patients who developed thrush maintained genetically similar strains throughout the study period, with minor genetic variations in all patients except one . Forty-six percent of these patients had either multiple strains throughout the study period (n = 2), strain replacement (n = 1), or species replacement (n = 2) . Of the patients who had multiple strains, one (I4) was infected by two different strains of Candida dubliniensis distinguished by a recently developed species-specific probe . These results suggest that commensal strains colonizing HIV-positive individuals can undergo alterations prior to producing an episode of thrush.

J Hosp Infect, 2002 Jan, 50(1), 66 - 75
Testing chemical germicides against Candida species using quantitative carrier and fingerpad methods; Traore O et al.; Six disinfectants were tested against Candida albicans, C . parapsilosis and C . tropicalis using quantitative carrier tests based on glass (QCT-1) and metal (QCT-2) surfaces . C . albicans was also used to test four topical agents by a fingerpad method . Hard water (200 ppm as CaCO(3)) was the product diluent . In preliminary tests with QCT-1 and QCT-2, the testing was with or without a soil load; subsequent tests and fingerpad tests included soil . In QCT-1 and QCT-2, each carrier received 10 microL (5.0 x 10(6) - 1.0 x 10(7)colony forming units) of Candida, and was air dried for 1 h, then exposed to 1 mL or 50 microL of test product at 22 +/- 2 degrees C for up to 10 min . Controls received an equivalent volume of saline . For fingerpad tests, each digit received 10 microL of inoculum, which was allowed to dry and exposed to 1 mL of test product for 20 s . Inoculated plates of Sabouraud's dextrose agar were held for 48 h at 30 degrees C and colonies counted to determine reductions in colony forming units . In tests on both hard surfaces and fingerpads, ethanol and products based on ethanol reliably and rapidly inactivated all the Candida species tested . Products with sufficient potency to have tuberculocidal claims produced substantial reductions in the titre of C . albicans, although some showed a lesser reduction in titre of C . tropicalis and C . parapsilosis . This may reflect differences in cell hydrophobicity between Candida species, and highlights the need for care in selecting a suitable surrogate for disinfectant tests . The quantitative carrier and fingerpad protocols are suitable for assessing the activity of disinfectants and topical antiseptics against candida .

J Dent Res, 2002 Jan, 81(1), 28 - 32
Saliva promotes Candida albicans adherence to human epithelial cells; Holmes AR et al.; Adhesion of Candida cells to oral surfaces is an initial event in pathogenesis . Since specific immobilized salivary components mediate the binding of Candida albicans to hydroxyapatite, we hypothesized that saliva may also promote adherence to oral epithelia via a similar mechanism . In an in vitro model, C . albicans ATCC 10261 yeast cells adhered in a saturable manner to monolayers of three cultured human epithelial cell lines (A549, HEp-2, and HET-1A) . The addition of whole saliva to the assay promoted the binding of C . albicans to all cell lines in a dose-dependent manner, but pre-incubation of the epithelial cells with pooled whole saliva had no effect on subsequent adherence . Pre-incubation of the yeast cells with pooled whole saliva, however, significantly enhanced (by up to 120%, P < 0.05) binding to epithelial cell monolayers, and pooled saliva that had been pre-incubated with C . albicans yeast cells was defective in promoting yeast adherence . There was a negative correlation (r = 0.68, P < 0.005) between specific IgA titers against whole cells of C . albicans and adherence-promoting activities for individual saliva samples . The adhesion-inhibitory effect of specific anti-C . albicans IgA was reversed by depletion of IgA from saliva by affinity chromatography . Factors in whole saliva, therefore, bound to the yeast cells, counter the C . albicans-specific salivary IgA inhibitory effect on adhesion and promote the adherence of C . albicans yeast cells to cultured epithelial cells.

Arch Pharm (Weinheim), 2001 Nov, 334(11), 361 - 5
Synthesis and antimicrobial activities of 5-fluoro-1,2,6-trisubstituted benzimidazole carboxamide and acetamide derivatives; Kus C et al.; Some 5-fluoro-6-substitute-1 H-benzimidazole-2-carbamates (12a-e), 5-fluoro-6-substituted 1H-benzimidazole-2-acetate (13a-e) and 2-acetamide (14a-f) derivatives, 2-acetamido-5-fluoro-6-(morpholin-4-yl)-1-propyl-1H-benzimidazole (15), and 1-cyclopropyl-2-ethyl-5-fluoro-6-(4-methylpiperazin-1-yl)- 1H-benzimidazole (16) were synthesized, and their antimicrobial and antifungal activities evaluated . Compound 12c exhibited the best activity against Candida albicans.

Eur J Ophthalmol, 2001 Oct-Dec, 11(4), 380 - 2
Candida keratitis in a patient with candidiasis of the fingernails; Jradeh EF et al.; PURPOSE: To report a case of Candida keratitis following penetrating keratoplasty in a patient with Candidiasis of the fingernails . CASE REPORT: An 80 year-old male presented with recurrent Candida keratitis following penetrating keratoplasty . Patient was found to have evidence of Candidiasis of the fingernails . COMMENT: Candidiasis of the fingernails have led to recurrent fungal keratitis following penetrating keratoplasty . Cultures grew Candida albicans . Preoperative recognition, prompt and appropriate therapy of foci of infection may prevent Candida keratitis following penetrating keratoplasty.

Mycoses, 2001 Dec, 44(11-12), 446 - 9
Trends in species causing fungaemia in a tertiary care medical centre over 12 years; Malani PN et al.; Trends in the species of yeast causing fungaemia over a 12-year period at a large tertiary care medical centre were reviewed . A total of 966 unique episodes of fungaemia occurred in 898 patients . There was an overall trend toward fewer fungaemic episodes due to Candida albicans and more due to Candida glabrata and Candida parapsilosis . However, C . albicans remained the predominant species causing fungaemia, and the proportion due to other species varied from year to year . Candida glabrata was disproportionately isolated from older adults, whereas C . parapsilosis was common among neonates and infants . The trends of increasing isolation of C . glabrata and decreasing isolation of C . albicans were associated with increasing usage of fluconazole, but changes in the proportion of fungaemias due to other species appeared to have no association with fluconazole usage.

Mycoses, 2001 Dec, 44(11-12), 437 - 45
Candida africana sp . nov., a new human pathogen or a variant of Candida albicans?
Tietz HJ, Hopp M, Schmalreck A, Sterry W, Czaika V.
Atypical Candida strains were isolated from patients in Madagascar, Angola and Germany . These isolates were slow growing and were unable to produce chlamydospores . They had atypical carbohydrate assimilation profiles . All strains were unable to assimilate the amino sugars N-acteylglucosamine and glucosamine as well as the disaccharide trehalose and the organic acid DL-lactate . They were germ-tube-positive in serum, but only some of these organisms produced pseudohyphae after a long incubation . As shown by Fourier transform infrared spectroscopy the atypical Candida isolates clustered as a monophyletic group different from C . albicans and C . dubliniensis . All strains belonged to C . albicans serotype B . Considering all data presented here, this group of Candida strains differs from any other known member of the genus Candida . Therefore, it is suggested to represent a new species within the genus Candida for which the name Candida africana is proposed.

Dent Update, 2001 Apr, 28(3), 132 - 9
Oral candidosis; McIntyre GT; Oral candidoses are frequently encountered in the practice of dentistry . Although most oral candidoses are symptomless, the can indicate the presence of an underlying systemic disease, and the persistence of oral candidosis following appropriate conventional management may be one of the first signs of undiagnosed immunosuppression . The opportunistic pathogen Candida albicans is the most commonly isolated species from oral candidal lesions; however, the non-albicans Candida spp . are also implicated in the aetiology of oral candidoses . The effective management of oral candidosis is dependent on an accurate diagnosis, identification and elimination of any predisposing factors (where possible), and the prescription of either topical or systemic antifungal agents . Oral candidosis may have significant implications for the general health of immunosuppressed patients, particularly when caused by the non-albicans spp . and, in cases of severe immunosuppression, systemic candidosis can be life-threatening . This article outlines the clinical presentation and appropriate management for the commonly presenting oral candidal conditions.

Rev Gastroenterol Peru, 2001 Oct-Dec, 21(4), 287 - 99
{Endoscopic findings in HIV-infected patients with esophageal symptoms . Experience at the Cayetano Heredia National Hospital}; Narvarte G et al.; This study was done in patients with HIV infection and upper digestive symptoms as odinophagia, dysphagia and/or retrosternal pain who attended the Cayetano Heredia National Hospital in Lima, Peru . Those included in the study had an upper endoscopy and a CD4 count . Also previous opportunistic infections were determined . Samples were taken from the oropharyngeal cavity and sent for direct exam and culture . During endoscopy, photos were taken from the upper, middle, and lower third of the esophagus and the esophageal compromise was classified . Biopsies and brushings samples were obtained and sent for direct exam, histopathology and culture . RESULTS: 751 patients with HIV infection attended the Cayetano Heredia National Hospital between May 1996 and June 1999, 83 were included due to esophageal symptoms . Male / Female ratio : 4/1, mean age : 30.95 +/- 9.87 . Cultures positive for Candida: 84.30% of esophageal biopsies samples, 88% of esophageal brushings and 60.2% of oropharyngeal cavity . The C . albicans species was isolated in 95.7% of biopsies, 93% of brushings and 96% of oropharyngeal cavity . The most common endoscopic finding was white plaques (71%), endoscopic grade 3 (36.1%) . The most frequent previous opportunistic infection was PCP pneumonia followed by TBC . The positive predictive value for white plaques in patients with dysphagia and odynophagia was 89.8% . CD4 count between 0 and 50 lymphocytes per mm(3) was seen in 70% of the cases . The mean of CD4 lymphocytes of these patients was lower (p< 0.01) when compared to the mean of patients in the control group with no symptoms . CONCLUSIONS: Candida albicans was the most common isolated pathogen in the esophagus in patients with HIV infection and with esophageal symptoms, a significant difference in CD4 count between these patients and the control group was determined.The positive predictive value for finding esophageal white plaques in patients dysphagia and odinophagia was high, and there was no difference between the average of CD4 count and the endoscopic grade of esophageal involvement.

Antonie Van Leeuwenhoek, 2001 Sep, 79(3-4), 297 - 309
Killing of Candida albicans by histatin 5: cellular uptake and energy requirement; Gyurko C et al.; Histatins, a group of histidine-rich proteins in human saliva, exhibit antimicrobial activity and are therefore considered to be important in the prevention of infections in the oral cavity . Although killing of C . albicans by histatins has been extensively studied, little is known about the processes responsible for this antifungal activity . Recent studies show the requirement of metabolic activity and ATP production for histatin 5 killing activity . Therefore, the goal of this study was to investigate the kinetics of histatin 5 interaction at different temperatures with C . albicans wild type cells and with respiratory deficient mutants of C . albicans . Synthetic histatin 5 was labeled with fluorescein-5-isothiocyanate (FITC) and its association with C . albicans cells was followed by epi-fluorescence microscopy and fluorescence confocal microscopy . At 37 degrees C, histatin 5 accumulates intracellularly, and both killing activity and uptake of unlabeled and FITC-labeled histatin 5 are time- and concentration-dependent . At 4 degrees C, no killing is observed and FITC-histatin 5 is only associated with the cytoplasmic membrane . Internalization and killing activity only occurs after cells are transferred to 37 degrees C . In addition, cellular accumulation of histatin 5 is concomitant with a moderate alteration of membrane integrity leading to the release of UV-absorbing cell components into the medium . The uptake of histatin 5, the release of UV-absorbing materials and killing of C . albicans are markedly decreased by the respiratory inhibitor sodium azide . Concomitantly, respiratory deficient mutants of C . albicans are also less susceptible to histatin 5 . These results indicated that histatin 5 killing activity could be directly correlated to histatin 5 internalization . Both of these processes are prevented by modulators of cellular metabolic activity.

Mol Vis, 2002 Jan 25, 8, 10 - 6
Evaluation of lectin staining in the diagnosis of fungal keratitis in an experimental rabbit model; Garcia ML et al.; PURPOSE: To assess the sensitivity, specificity, and reliability of peroxidase labeled lectin staining in the diagnosis of fungal keratitis in an experimental rabbit model . METHODS: Fungal keratitis by Candida albicans, Aspergillus fumigatus, and Fusarium solani was induced in rabbits . WGA-peroxidase staining of 660 corneal sections was performed . Fungal staining was evaluated independently by two observers . The test sensitivity, specificity, and reliability indexes were calculated . RESULTS: The sensitivity of the lectin staining test for Candida albicans was 100% (95% CI: 93.51-100.00), and specificity was 100% (95% CI: 93.51-100.00) . The sensitivity of the test for Aspergillus fumigatus was 96.36% (95% CI: 86.46-99.35), and specificity was 100% (95% CI: 93.51-100.00) . The sensitivity of the test for Fusarium solani was 96.36% (95% CI: 86.46-99.35) and specificity was 96.15% (95% CI: 85.74-99.31) . There was also a high degree of test-retest and inter-rater concordance for all three fungi tested . The test-retest k reliability indexes were 0.9455, 0.9636, and 0.8879, for Candida albicans, Aspergillus fumigatus, and Fusarium solani, respectively . The inter-rater k reliability indexes were 0.9636, 0.9818, and 0.9252, for Candida albicans, Aspergillus fumigatus, and Fusarium solani, respectively . CONCLUSIONS: WGA-peroxidase staining is a very sensitive, specific, and reliable test for the identification of fungi in an experimental rabbit model of fungal keratitis.

J Antimicrob Chemother, 2002 Feb, 49(2), 345 - 51
Comparison of Etest, chequerboard dilution and time-kill studies for the detection of synergy or antagonism between antifungal agents tested against Candida species; Lewis RE et al.; Currently, there is considerable debate regarding the best in vitro method for testing antifungal combinations against Candida spp . In this study, we compared the results obtained by chequerboard dilution, time-kill studies and Etest for several antifungal combinations against Candida spp . Three Candida albicans isolates (fluconazole MICs of 1.0, 32 and >256 mg/L) and three non-albicans Candida isolates (C . glabrata, C . tropicalis and C . krusei) were tested in RPMI 1640 medium . By chequerboard testing, the majority of antifungal combinations were found to be indifferent . Notably, antagonism was identified by time-kill studies and by Etest for combinations of amphotericin B-fluconazole, but it was not detected by the chequerboard method . Pre-exposure of isolates to fluconazole did not affect results of the Etest or chequerboard method, but it did increase the frequency of antagonism noted by time-kill methods . This study indicates that chequerboard dilution testing in RPMI medium may not reliably detect the attenuation of amphotericin B activity . Of the three methods, Etest was the simplest to use and yielded reproducible results for testing antifungal combinations.

Trends Mol Med, 2002 Feb, 8(2), 76 - 81
Molecular basis of resistance to azole antifungals; Lupetti A et al.; The increased incidence of invasive mycoses and the emerging problem of antifungal drug resistance has prompted investigations of the underlying molecular mechanisms, particularly for the azole compounds central to current therapy . The target site for the azoles is the ERG11 gene product, the cytochrome P450 lanosterol 14alpha-demethylase, which is part of the ergosterol biosynthetic pathway . The resulting ergosterol depletion renders fungal cells vulnerable to further membrane damage . Development of azole resistance in fungi may occur through increased levels of the cellular target, upregulation of genes controlling drug efflux, alterations in sterol synthesis and decreased affinity of azoles for the cellular target . Here, we review the adaptative changes in fungi, in particular Candida albicans, in response to inhibitors of ergosterol biosynthesis . The molecular mechanisms of azole resistance might help in devising more effective antifungal therapies.

J Infect Chemother, 2000 Dec, 6(4), 216 - 21
Antifungal susceptibility of 262 bloodstream yeast isolates from a mixed cancer and non-cancer patient population: is there a correlation between in-vitro resistance to fluconazole and the outcome of fungemia?
Kovacicova G, Krupova Y, Lovaszova M, Roidova A, Trupl J, Liskova A, Hanzen J, Milosovic P, Lamosova M, Macekova L, Szovenyiova Z, Purgelova A, Obertik T, Bille J, Krcmery V.
The aim of this study was to test the antifungal susceptibility of 262 bloodstream yeast isolates (164 Candida albicans strain, 88 non-albicans Candida spp . and 10 non-Candida yeasts) recovered from 169 surgical, neonatal, critically ill intensive care unit patients (ICU), and cancer patients (mixed patient population) to amphotericin B (AmB), fluconazole (FLU), 5-flucytosine (5-FC), itraconazole (ITRA), ketoconazole (KETO), miconazole (MICO), and nystatin (NYS), in order to correlate in-vitro resistance to fluconazole with the outcome of fungemia . The agar disk diffusion test was used to assess the susceptibility of the 262 bloodstream yeasts isolates . In addition, 78 strains isolated from cancer patients were also tested with the E-test . There were no differences in the susceptibility of the various C . albicans strains tested, except in 40 isolates from surgery patients, which showed a somewhat lower susceptibility to KETO and MICO to (3.7-5.5% resistance) . There were no C . albicans strains resistant to AmB, NYS, or FLU . There were slight differences in the susceptibility patterns of the 88 non-albicans Candida spp . (NAC) isolates . Resistance to AmB and NYS appeared in 1 strain of C . guillermondii (minimum inhibitory concentration; MIC to AmB; 4 microg/ml) and in 1 strain of C . parapsilosis (MIC to NYS, 8 microg/ml and MIC to AmB, 2 microg/ml) . All other NACs were susceptible to both polyenes (AmB and NYS) . Nine of the 11 strains of C . krusei were resistant to FLU (MIC >or= 64 microg/ml), the 2 exceptions showed, respectively, MICs for FLU of 6 and 32 microg/ml ("dose-dependent" susceptibility) . However, only 2 of 29 C . glabrata strains were fully FLU-resistant (MIC >or= 64 microg/ml), 27 being susceptible with MIC values of 0.5-8 microg/ml . Apart from 9 C . krusei and 2 C . glabrata strains, 2 C . parapsilosis strains and 1 strain of C . tropicalis were also FLU-resistant . Among the 88 NACs, 17.04% were FLU-resistant and 3.7% were KETO- and ITRA-resistant . Resistance to 5-FC and AmB was minimal . We compared the outcomes of patients infected with FLU-resistant vs FLU-susceptible yeasts in 161 evaluable patients treated with FLU . Attributable mortality was significantly higher (19.0% vs 8.6%; P < 0.01) in patients infected with the FLU-resistant yeasts.

J Infect Chemother, 2000 Sep, 6(3), 151 - 4
Synergistic effect of ofloxacin and fluconazole against azole-resistant Candida albicans; Sasaki E et al.; We investigated the combination effects of ofloxacin and fluconazole against azole-resistant Candida albicans strains in vitro and in vivo . Ofloxacin alone showed no efficacy against the azole-resistant C . albicans strain, C26 . The in-vitro combination effects were evaluated by the checkerboard method, calculated as the fractional inhibitory concentration (FIC) index, but there was no synergistic effect of the combination . The activity of the drug efflux pump in the azole-resistant C . albicans strains was measured by intracellular rhodamine 6G concentration . When the cells were incubated with ofloxacin or grepafloxacin, the intracellular rhodamine 6G concentration was significantly increased in the azole-resistant C . albicans strain . In-vivo combination effects were evaluated in murine disseminated candidiasis . The survival of the mice was not prolonged, but counts of the yeast cells in the kidney and spleen were reduced following treatment with the combination of ofloxacin (20 mg/kg) and fluconazole (20 mg/kg) . The combination of ofloxacin and fluconazole may represent an effective strategy to treat infections caused by azole-resistant C . albicans.

J Infect Chemother, 2000 Sep, 6(3), 144 - 7
Effects of fluconazole on viable cell count in experimental intraperitoneal Candida abscesses; Mikamo H et al.; We investigated the effects of fluconazole in experimental intraperitoneal Candida abscesses in neutropenic mice treated with cyclophosphamide to assess a clinically appropriate method for the application of fluconazole in fungal infections in patients with neutropenia . The efficacy of fluconazole in fungal infection was investigated in treatments started immediately after Candida albicans inoculation and before C . albicans inoculation (preventive use of fluconazole) . In this intraabdominal fungal abscess model, it was confirmed that fungi in already formed abscess were not reduced in number by fluconazole, even when those fungi were susceptible to fluconazole . Fluconazole was effective with both administration methods . We conclude from this study that the effect of the preventive use of fluconazole was equivalent to the effect when treatment was started immediately after fungal inoculation.

J Infect Chemother, 1999 Sep, 5(3), 163 - 167
Candida glabrata fungemia in a tertiary cancer institution in Slovakia; Krcmery Jr V V et al.; Because of controversial data on virulence and mortality, six cases of fungemia caused by Candida glabrata were reviewed in a single cancer institution within 8 years . Risk factors and outcome of C . glabrata, Candida albicans, and other non-albicans Candida spp . appearing within the same period under the same antibiotic policy at the same institution were compared . Among other non-albicans Candida spp . in 1990-1997 C . glabrata fungemias showed a decreasing tendency, from 9% to 4.5% in 1997 . Analyzing the proportion of C . glabrata among blood cultures, among 170 positive blood cultures 12 were caused by C . glabrata (6.2% among all pathogens and 24% among non-albicans Candida spp.) . C . glabrata among all fungemias was diagnosed as the fourth most common pathogen after C . albicans, C . krusei, and C . parapsilosis . Three of six C . glabrata fungemias were breakthrough . Two appeared during prophylaxis with itraconazole and one during fluconazole prophylaxis . Five of six received broadspectrum antibiotic therapy with third-generation cephalosporines, five of six had vascular catheter insertion, four of six were neutropenic, and two of six received amphotericin B therapy . One patient died before his blood cultures were reported to be positive . Overall mortality of C . glabrata fungemia was 16.7% . One patient died of underlying disease with fungemia . There were no significant differences in risk factors between C . glabrata and C . albicans . However, overall and crude mortality was lower in C . glabrata than in C . albicans (25.5% vs . 16.7%; P = 0.03) . Attributable mortality was lower in comparison to C . albicans (0 vs . 15.7% in C . albicans; P = 0.001).

Mol Genet Genomics, 2001 Dec, 266(4), 624 - 31 Epub 2001 Oct 11.
Spontaneous second-site suppressors of the filamentation defect of prr1Delta mutants define a critical domain of Rim101p in Candida albicans; Porta A et al.; In response to changes in ambient pH the opportunistic pathogen Candida albicans differentially expresses a number of genes . The response to pH affects morphological differentiation and virulence . The pathway controlling the pH response terminates in the zinc-finger containing transcription factor encoded by RIM101/PRR2 . By analogy to the pH response pathway of Aspergillus nidulans, PRR1 of C . albicans encodes a protein that is presumably required to convert Rim101p from an inactive to an active form by proteolytic removal of a C-terminal peptide . A prr1Delta mutant is compromised in its ability to differentiate into the filamentous form . Spontaneous phenotypic revertants of a prr1Delta mutant were selected by their ability to form filamentous colonies . These mutants were also found to be defective in pH-dependent gene expression . Each of the eight mutants examined contained a heterozygous dominant mutation at the RIM101 locus . This was demonstrated genetically in all of the mutants, and directly by sequence determination of both alleles in two of the mutants . The mutant alleles conferred the ability to filament to a prr1Delta mutant, thus demonstrating that they were directly responsible for suppressing the filamentation defect . Seven of the mutant alleles contained a 1-bp substitution and one contained two substitutions at adjacent positions . The mutations were clustered within a 90-bp region near the 3'-end of the gene . In all cases the mutation generated a nonsense codon that resulted in premature termination of Rim101p; the mutant proteins were truncated by 75-104 amino acids . The results define a critical region in the C-terminal region of Rim101p and are consistent with the proposed proteolytic activation of Rim101p.

Mol Biol Cell, 2002 Jan, 13(1), 134 - 45
Hyphal elongation is regulated independently of cell cycle in Candida albicans; Hazan I et al.; The mechanism for apical growth during hyphal morphogenesis in Candida albicans is unknown . Studies from Saccharomyces cerevisiae indicate that cell morphogenesis may involve cell cycle regulation by cyclin-dependent kinase . To examine whether this is the mechanism for hyphal morphogenesis, the temporal appearance of different spindle pole body and spindle structures, the cell cycle-regulated rearrangements of the actin cytoskeleton, and the phosphorylation state of the conserved Tyr19 of Cdc28 during the cell cycle were compared and found to be similar between yeast and serum-induced hyphal apical cells . These data suggest that hyphal elongation is not mediated by altering cell cycle progression or through phosphorylation of Tyr19 of Cdc28 . We have also shown that germ tubes can evaginate before spindle pole body duplication, chitin ring formation, and DNA replication . Similarly, tip-associated actin polarization in each hypha occurs before the events of the G(1)/S transition and persists throughout the cell cycle, whereas cell cycle-regulated actin assemblies come and go . We have also shown that cells in phases other than G(1) can be induced to form hyphae . Hyphae induced from G(1) cells have no constrictions, and the first chitin ring is positioned in the germ tube at various distances from the base . Hyphae induced from budded cells have a constriction and a chitin ring at the bud neck, beyond which the hyphae continue to elongate with no further constrictions . Our data suggest that hyphal elongation and cell cycle morphogenesis programs are uncoupled, and each contributes to different aspects of cell morphogenesis.

J Nat Prod, 2002 Jan, 65(1), 62 - 4
Antifungal activity of benzoic acid derivatives from Piper lanceaefolium; Lopez A et al.; Bioactivity-guided fractionation of a methanol extract from the leaves of Piper lanceaefolium resulted in the isolation of four new benzoic acid derivatives (1-4), together with taboganic acid, pinocembrin, and pinocembrin chalcone . Lanceaefolic acid methyl ester (3) and pinocembrin chalcone displayed activity against Candida albicans with a minimal inhibitory concentration value of 100 microg/mL in both cases.

Genetics, 2002 Jan, 160(1), 37 - 48
The Ess1 prolyl isomerase is required for growth and morphogenetic switching in Candida albicans; Devasahayam G et al.; Prolyl-isomerases (PPIases) are found in all organisms and are important for the folding and activity of many proteins . Of the 13 PPIases in Saccharomyces cerevisiae only Ess1, a parvulin-class PPIase, is essential for growth . Ess1 is required to complete mitosis, and Ess1 and its mammalian homolog, Pin1, interact directly with RNA polymerase II . Here, we isolate the ESS1 gene from the pathogenic fungus Candida albicans and show that it is functionally homologous to the S . cerevisiae ESS1 . We generate conditional-lethal (ts) alleles of C . albicans ESS1 and use these mutations to demonstrate that ESS1 is essential for growth in C . albicans . We also show that reducing the dosage or activity of ESS1 blocks morphogenetic switching from the yeast to the hyphal and pseudohyphal forms under certain conditions . Analysis of double mutants of ESS1 and TUP1 or CPH1, two genes known to be involved in morphogenetic switching, suggests that ESS1 functions in the same pathway as CPH1 and upstream of or in parallel to TUP1 . Given that switching is important for virulence of C . albicans, inhibitors of Ess1 might be useful as antifungal agents.

Biomaterials, 2002 Mar, 23(5), 1327 - 35
Synthesis and characterization of novel water soluble amphotericin B-arabinogalactan conjugates; Ehrenfreund-Kleinman T et al.; The coupling of amphotericin B (AmB), a water-insoluble antifungal agent, to arabinogalactan (AG) via an imine or amine bond was systematically investigated . AG was oxidized using potassium periodate, purified from the oxidizing agent using ion-exchange chromatography, and reacted with AmB to form the Schiff base . The Schiff base was reduced to the amine using borohydride . All reactions took place in aqueous media . The purification of the oxidized AG from the oxidizing agent was essential to prevent oxidative degradation of AmB at the coupling step . We investigated the effects of AmB to AG ratio, buffer type, and reaction pH on the reaction yield, molecular weight, conjugate activity against pathogenic yeast and hemolytic activity . The optimum coupling conditions were buffer borate 0.1 M, pH 11 at room temperature for 48 h . Lower toxicity in vivo was achieved by using low-pressure gel permeation chromatography and applying the solution of AmB-AG conjugate through a Sephadex column . Both amine and imine AmB-AG conjugates were soluble in water and exhibited improved stability in aqueous solutions as compared to the unbound drug . The conjugates showed comparable minimum inhibitory concentration (MIC) values against Candida albicans . The conjugates were about 60 times less hemolytic against sheep erythrocytes than the free drug, and about 40 times less toxic in BALB/c mice.

Eur J Pharm Sci, 2002 Feb, 15(1), 49 - 61
Synthesis, structure investigations, and antimicrobial activity of selected s-trans-6-aryl-4-isopropyl-2-{2-{(E)-1-phenylalkylidene}-(E)-hydrazino}-1,4-dihydropyrimidine hydrochlorides; Gossnitzer E et al.; A series of 6-aryl-4-isopropyl-2-{2-(1-phenylalkylidene)hydrazino}-1,4-dihydropyrimidine hydrochlorides was prepared and tested for antibacterial and antifungal effects . The title compounds were synthesized by cyclocondensation of N(2)-(1-phenylalkylideneamino)guanidines with 1-aryl-4-methyl-2-penten-1-ones . Structure analyses of the prepared compounds were accomplished by means of 1D and 2D NMR spectroscopy . The analyses showed that the ring closure reaction took place regioselectively in all cases and generated exclusively 2-(phenylalkylidenehydrazino)dihydropyrimidines . According to the NOE experiments, the applied N(2)-(1-phenylalkylideneamino)guanidines exist in {D(6)}DMSO solution as s-trans-(E)- and the title compounds as s-trans-(E,E)-isomers . The antimicrobial activity was evaluated against representative Gram-negative and Gram-positive bacteria, and against the pathogenic yeast Candida albicans . The tested title compounds showed weak antibacterial activity against Gram-positive bacteria, and one compound was active against Candida albicans.

Allerg Immunol (Paris), 2001 Dec, 33(10), 388 - 94
{Treatment of sino-nasal polyposis by Candida albicans immunotherapy: apropos of 4 cases}; Benoliel P; The NSP is an inflammatory chronic disease of the mucous of nose and sinuses . None etiological treatment is known up to now . The aim of this study is to consider a model of autoallergy as etiology for NSP proven by specific immunotherapy (STI) to Candida albicans (CA) . METHODS: Four NSP treated by SIT to Candida albicans are reported . The patients are treated either by subcutaneous injections or sublingual drops . The frequency is one injection per week or a few drops per day (absorbed extract on calcium phosphate or aqueous Stallergenes) . RESULTS: The cumulated doses varies from 465 Index of Concentration (IC) to 117500 IC on a period of 3 to 4 years . The results are evaluated according the rhino-sinusal semeiology, the intensity of symptoms, and the stage of polyposis . The SIT is also active on both a late and an immediate components for the symptoms, and the cutaneous tests . The results are significant 60% to 80% of improvement . The viral or bacterial infections reactivate both types of hypersensitivity and they are prevented by SIT . The nasal hyperactivity observed as a more advanced non specific stage of the PNS is also improved by ITS . In two of the clinical cases, the pollenogenic seasonal obstruction is added to the nasal perennial obstruction in a sharp manner . The pollenogenic allergy is also improved after SIT to CA without any other associated SIT . CONCLUSION: The model of autoallergy already proven as etiology for atopic dermatitis can serve as a base of exploration of PNS . That is showing the presence of IgE antibody corresponding to intracellular proteinic autoallergens having an analogy to environment allergens . The allergy to Candida albicans can thus be considered as an etiology of the PNS.

J Ethnopharmacol, 2002 Feb, 79(2), 155 - 63
Further evaluation of Rwandan medicinal plant extracts for their antimicrobial and antiviral activities; Cos P et al.; A total of 45 Rwandan plant extracts, belonging to 37 different plant species out of 21 families, were investigated for their antibacterial, antifungal, and antiviral properties . The plants were selected on the base of their ethnomedicinal use against infections and autoimmune diseases . From all the plant extracts tested, only Clematis hirsuta (leaves) showed a pronounced antifungal activity against Candida albicans and the dermatophytes Trichophyton rubrum, Epidermophyton floccosum, and Microsporum canis . Seven plant extracts showed a high antiviral activity against the DNA-virus Herpes simplex type 1, while five and three plant extracts were highly active against the RNA-viruses Coxsackie and Polio, respectively . Only Macaranga kilimandscharica (leaves) showed an interesting anti-measles activity, whereas Eriosema montanum (leaves) and Entada abyssinica (leaves) were highly active against Semliki forest virus . Some plant extracts showed an antibacterial activity against Gram-positive bacteria and Mycobacterium fortuitum, but none of them were active against the Gram-negative bacteria tested.

Acta Trop, 2002 Feb, 81(2), 101 - 10
Candida commensalism and virulence: the evolution of phenotypic plasticity; Soll DR; Candida albicans and related species live as benign commensals in one or more body locations in a majority of healthy individuals . As opportunistic pathogens, they are poised to overgrow cavities and penetrate tissue in response to an alteration in host physiology that presumably compromises the immune functions that normally suppress their growth . There is little evidence of the emergence of successful drug-resistant or hypervirulent strains that predominate in either the commensal or disease states . It appears more likely that all strains possess similar capabilities for rapid adaptation to drug therapy, the immune response and changes in body location and in host physiology . It is suggested that the mechanisms for rapid adaptation lie in the developmental programs of the bud-hypha transition and high frequency phenotypic switching, and in the modulation of the expression of virulence genes in response to environmental cues.

Med Mycol, 2001, 39 Suppl 1, 87 - 100
Signal transduction pathways and cell-wall construction in Candida albicans; Navarro-Garcia F et al.; Signal transduction pathways are the molecular mechanisms responsible for detecting and transmitting changes in the surrounding environment to the nucleus where appropriate responses are generated . The cell wall is the most external structure of the fungal cell and, in pathogenic fungi, is responsible for specifically interacting with the mammalian host cell in a highly dynamic interplay . Recent work has shown the role that some signal transduction pathways, involving members of the MAP kinase family, have in this process in the nonpathogenic model organism Saccharomyces cerevisiae . However, as yet little is known about these phenomena in pathogenic fungi . The aim of this review is to characterize the existing signal transduction pathways in Candida albicans and their relationship with the cell-wall construction.

Med Mycol, 2001, 39 Suppl 1, 75 - 86
N-glycosylation of yeast, with emphasis on Candida albicans; Cutler JE; Fungal cell wall N-linked glycans have been studied most extensively in Saccharomyces cerevisiae and in Candida albicans . The glycans are located on the fungal cell surface in the form of phosphomannoprotein complexes and the amount of glycosylation is influenced both by genetics and environmental factors . The glycans, which are comprised mostly of mannan, are important in fungal-host interactions, as they make first contact with the immune system . Initial N-linked glycosylation events take place in the endoplasmic reticulum and are conserved throughout all eukaryotes, but yeasts are capable of additional glycosylation that may result in a glycan comprised of more than 200 mannose units . In C . albicans, the glycan can be delineated into an inner mannan core, which is similar to mammalian glycoproteins, an alpha-linked mannan backbone with alpha-oligomannosyl side chains, and beta(1,2)-oligomannosides which are phosphodiester linked to the alpha-mannan . Both the beta-oligomannosides, which make up the acid-labile part of the phosphomannan complex, and alpha-oligomannosides, which make up the acid-stable part of the complex, serve as adhesins in the attachment of C . albicans yeast cells to host splenic and lymph node macrophages . The beta-oligomannosides can induce release of tumour necrosis factor (TNF)-alpha, and antibodies specific to certain beta-oligomannosides enhance host resistance to various forms of candidiasis . The importance of the N-linked glycans in fungal-host interactions provides rationale for further studies, which may well lead to effective immunotherapeutic strategies for prevention and, possibly, treatment of disease.

Med Mycol, 2001, 39 Suppl 1, 67 - 74
O-glycosylation; Ernst JF et al.; O-Glycosylation in many fungal species is initiated in the endoplasmic reticulum by protein mannosyltransferases (Pmt-proteins), which transfer mannose to serine or threonine residues, and it is completed by mannosyltransferases (Mnt-proteins) in the Golgi . In this review, some recent results on O-glycosylation in the human fungal pathogen Candida albicans are discussed and compared to the corresponding knowledge in the non-pathogenic yeast Saccharomyces cerevisiae . The Pmt-family in C . albicans comprises five isoforms, of which Pmt1p and Pmt6p have been studied in detail . Surprisingly, O-glycosylation mediated by Pmt-proteins is required not only for the modification of several secreted and cell-wall proteins, but also affects yeast-hyphal morphogenesis (dimorphism) and resistance to several antifungal compounds . Furthermore, Pmt1- and Pmt6p-activities maximize adherence to host cells and determine or contribute to virulence in models of systemic infection . Thus, O-glycosylation processes directly and/or indirectly affect several virulence traits of C . albicans and can be considered as potential antifungal targets.

Med Mycol, 2001, 39 Suppl 1, 1 - 8
Molecular organization of the cell wall of Candida albicans; Klis FM et al.; We have recently presented a molecular model of the cell wall of Saccharomyces cerevisiae . Here we discuss the evidence that a similar model is also valid for Candida albicans . We further discuss how cell-wall proteins are linked to the skeletal layer of the wall, and their potential functions . We emphasize that the composition and structure of the cell wall depends on growth conditions . Finally, cell-wall damage seems to activate a salvage mechanism resulting in restructuring of the cell wall.

J Allergy Clin Immunol, 2002 Jan, 109(1), 102 - 5
Downregulation of IFN-gamma production in patients with recurrent vaginal candidiasis; Carvalho LP et al.; BACKGROUND: Recurrent vaginal candidiasis (RVC) is an important health problem with unknown pathogenesis . Although impairment of the T-cell response is associated with persistent or recurrent candidiasis, data on immunologic responses in patients with RVC are controversial . OBJECTIVES: To evaluate the T-cell response in patients with RVC and the ability of cytokines and cytokine antagonists to modulate IFN-gamma production in cultures stimulated with Candida albicans antigens . METHODS: Participants in the study included 13 patients with RVC and 7 control women with sporadic candidiasis . Cytokines were determined by ELISA in supernatants of mononuclear cells with C albicans, purified protein derivative, or tetanus toxoid antigen . RESULTS: IFN-gamma production was absent or low in 11 of 13 women (84.6%) with RVC . Absent or low IFN-gamma production was specific to C albicans antigens (189 +/- 389 pg/mL), because high IFN-gamma levels were found in cultures stimulated with purified protein derivative (739 +/- 774 pg/mL) or tetanus toxoid antigens (1085 +/- 546 pg/mL) . Monoclonal antibody anti-IL-10 enhanced IFN-gamma levels (750 +/- 753 pg/mL), and IL-10 suppressed this cytokine production in patients with sporadic candidiasis . CONCLUSIONS: Mononuclear cells from patients with RVC stimulated with C albicans antigen have low or absent IFN-gamma production . IL-10 plays an important role in downregulation of the T-cell response in these patients.

Med Mycol, 2001 Dec, 39(6), 495 - 507
Plasminogen-binding activity of enolase in the opportunistic pathogen Pneumocystis carinii; Fox D et al.; The glycolytic enzyme enolase is one of the most abundant proteins expressed in fungi and has been shown to be an immunodominant cell-wall-associated antigen of the pathogenic fungus, Candida albicans . Enolase has also been found on the surface of some mammalian cells where it functions as a plasminogen-binding motif and facilitator of plasminogen activation to plasmin . To investigate the immunogenicity of enolase in the opportunistic pathogen, Pneumocystis carinii, the genomic and complementary DNA (cDNA) enolase were cloned and characterized . The predicted protein comprises 433 amino-acid residues and shows extensive homology to other fungal enolases, including those of C . albicans (76%), Aspergillus oryzae (79%) and Saccharomyces cerevisiae (77%) . The purified recombinant P . carinii enolase was immunogenic, and may be an important antigen and indicator of P . carinii infection . The active site and conformation metal ion-binding site residues necessary for dimerization and enzyme function are conserved in the predicted P . carinii enolase protein . Enolase of P . carinii is unique among the fungal enolases in that it possesses a catalytic carboxyl-terminal lysyl residue that was necessary and sufficient for the plasminogen-binding activity of the enolase of P . carinii . The activity of the plasminogen binding suggests its involvement in the local regulation of fibrinolysis within the alveolar space.

Infect Immun, 2002 Feb, 70(2), 985 - 7
Deletion of the two-component histidine kinase gene (CHK1) of Candida albicans contributes to enhanced growth inhibition and killing by human neutrophils in vitro; Torosantucci A et al.; We have observed that human neutrophils (polymorphonuclear leukocytes {PMNs}) have an increased growth-inhibitory and killing effect on a strain of Candida albicans with a deletion of CHK1, a gene encoding a putative histidine kinase . The PMN effect was not due to increased phagocytosis of the null strain . This observation may partially explain the reduced virulence in a hematogenously disseminated murine model of candidiasis.

Infect Immun, 2002 Feb, 70(2), 921 - 7
Transcriptional regulators Cph1p and Efg1p mediate activation of the Candida albicans virulence gene SAP5 during infection; Staib P et al.; The opportunistic fungal pathogen Candida albicans can cause superficial as well as systemic infections . Successful adaptation to the different host niches encountered during infection requires coordinated expression of various virulence traits, including the switch between yeast and hyphal growth forms and secretion of aspartic proteinases . Using an in vivo expression technology that is based on genetic recombination as a reporter of gene activation during experimental candidiasis in mice, we investigated whether two signal transduction pathways controlling hyphal growth, a mitogen-activated protein kinase cascade ending in the transcriptional activator Cph1p and a cyclic AMP-dependent regulatory pathway that involves the transcription factor Efg1p, also control expression of the SAP5 gene, which encodes one of the secreted aspartic proteinases and is induced by host signals soon after infection . Our results show that both transcriptional regulators are important for SAP5 activation in vivo . SAP5 expression was reduced in a cph1 mutant, although filamentous growth in infected tissue was not detectably impaired . SAP5 expression was also reduced, but not eliminated, in an efg1 null mutant, although this strain grew exclusively in the yeast form in infected tissue, demonstrating that in contrast to in vitro conditions, SAP5 activation during infection does not depend on growth of C . albicans in the hyphal form . In a cph1 efg1 double mutant, however, SAP5 expression in infected mice was almost completely eliminated, suggesting that the two signal transduction pathways are important for SAP5 expression in vivo . The avirulence of the cph1 efg1 mutant seemed to be caused not only by the inability to form hyphae but also by a loss of expression of additional virulence genes in the host.

Infect Immun, 2002 Feb, 70(2), 724 - 31
Primary role for CD4(+) T lymphocytes in recovery from oropharyngeal candidiasis; Farah CS et al.; Oropharyngeal candidiasis is associated with defects in cell-mediated immunity and is commonly seen in human immunodeficiency virus positive individuals and AIDS patients . A model for oral candidiasis in T-cell-deficient BALB/c and CBA/CaH nu/nu mice was established . After inoculation with 10(8) Candida albicans yeasts, these mice displayed increased levels of oral colonization compared to euthymic control mice and developed a chronic oropharyngeal infection . Histopathological examination of nu/nu oral tissues revealed extensive hyphae penetrating the epithelium, with polymorphonuclear leukocyte microabscess formation . Adoptive transfer of either naive or immune lymphocytes into immunodeficient mice resulted in the recovery of these animals from the oral infection . Reconstitution of immunodeficient mice with naive CD4(+) but not CD8(+) T cells significantly decreased oral colonization compared to controls . Interleukin-12 and gamma interferon were detected in the draining lymph nodes of immunodeficient mice following reconstitution with naive lymphocytes . This study demonstrates the direct requirement for T lymphocytes in recovery from oral candidiasis and suggests that this is associated with the production of cytokines by CD4(+) T helper cells.

Infect Immun, 2002 Feb, 70(2), 577 - 83
Cytokine and chemokine production by human oral and vaginal epithelial cells in response to Candida albicans; Steele C et al.; Oropharyngeal and vaginal candidiases are the most common forms of mucosal fungal infections and are primarily caused by Candida albicans, a dimorphic fungal commensal organism of the gastrointestinal and lower female reproductive tracts . Clinical and experimental observations suggest that local immunity is important in host defense against candidiasis . Accordingly, cytokines and chemokines are present at the oral and vaginal mucosa during C . albicans infections . Since mucosal epithelial cells produce a variety of cytokines and chemokines in response to microorganisms and since C . albicans is closely associated with mucosal epithelial cells as a commensal, we sought to identify cytokines and/or chemokines produced by primary oral and vaginal epithelial cells and cell lines in response to C . albicans . The results showed that proinflammatory cytokines were produced by oral and/or vaginal epithelial cells at various levels constitutively with considerable interleukin-1alpha (IL-1alpha) and tumor necrosis factor alpha, but not IL-6, produced in response to C . albicans . In contrast, Th1-type (IL-12 and gamma interferon) and Th2-type-immunoregulatory (IL-10 and transforming growth factor beta) cytokines and the chemokines monocyte chemoattractant protein 1 and IL-8 were produced in low to undetectable concentrations with little additional production in response to C . albicans . Taken together, these results indicate that cytokines and chemokines are variably produced by oral and vaginal epithelial cells constitutively, as well as in response to C . albicans, and are predominated by proinflammatory cytokines.

Infect Immun, 2002 Feb, 70(2), 517 - 27
Lymphocyte adhesion to Candida albicans; Forsyth CB et al.; Adherence of lymphocytes to the fungus is the first step in the direct lymphocyte-mediated antifungal effect against Candida albicans . In this study we identified macrophage-1 antigen (Mac-1) (CD11b/CD18, alpha(M)/beta(2)) as the lymphocyte surface structure responsible for the adhesion of activated lymphocytes to the hyphal form of the fungus . Antibodies specific for epitopes of the alpha-subunit (CD11b) and the beta(2)-subunit (CD18) of Mac-1 were shown to completely eliminate lymphocyte adhesion to C . albicans hyphae . Lymphocyte adhesion to C . albicans was also inhibited significantly by known ligands of Mac-1, including the extracellular matrix proteins laminin and fibrinogen, as well as engineered peptides containing arginine-glycine-aspartic acid sequences and the disintegrin echistatin . N-Acetyl-D-glucosamine and beta-glucan, which inhibit Mac-1-mediated adhesion to the yeast, blocked lymphocyte adhesion to hyphae . NIH 3T3 fibroblast transfectants expressing human CD11b/CD18 bound to C . albicans, and their binding was inhibited by antibodies specific for CD11b/CD18 . Finally, antibodies specific for CD11b/CD18 effectively inhibited the capacity of activated lymphocytes to have an antifungal effect against hyphae . Our results clearly identify Mac-1 (CD11b/CD18) as the lymphocyte surface structure that mediates activated lymphocyte adhesion to C . albicans and the resultant antifungal effect of the lymphocytes.

Gerodontology, 2001 Dec, 18(2), 73 - 8
Strain persistence of invasive Candida albicans in chronic hyperplastic candidosis that underwent malignant change; Williams DW et al.; OBJECTIVES: The aim of this study was to assess persistence and tissue invasion of Candida albicans strains isolated from a 65 year-old patient with chronic hyperplastic candidosis (CHC), that subsequently developed into squamous cell carcinoma (SCC) . MATERIALS AND METHODS: C . albicans (n=7) were recovered from the oral cavity of the patient over seven years . Confirmation of CHC and SCC in this patient was achieved by histopathological examination of incisional biopsy tissue . DNA fingerprinting was performed on the seven isolates from the CHC patient together with a further eight isolates from patients with normal oral mucosa (n=2), chronic atrophic candidosis (n=1), SCC (n=1) and CHC (n=4) . Genotyping involved the use of inter-repeat PCR using the eukaryotic repeat primer 1251 . Characterisation of the tissue invasive abilities of the isolates was achieved by infecting a commercially available reconstituted human oral epithelium (RHE; SkinEthic, Nice, France) . After 24 h, C . albicans tissue invasion was assessed by histopathological examination . RESULTS: DNA fingerprinting demonstrated strain persistence of C . albicans in the CHC patient over a seven year period despite provision of systemic antifungal therapy . The strain of C . albicans isolated from this patient was categorised as a high invader within the RHE compared to other isolates . CONCLUSIONS: Candidal strain persistence was evident in a patient with CHC over seven years . This persistence may be due to incomplete eradication from the oral cavity following antifungal therapy or subsequent recolonisation from other body sites or separate exogenous sources . The demonstration of enhanced in vitro tissue invasion by this particular strain may, in part, explain the progression to carcinoma.

J Prosthet Dent, 1995 Jun, 73(6), 530 - 3
In vitro study on the effects of trial denture cleansers with berberine hydrochloride; Nakamoto K et al.; The antifungal activity of trial denture cleansers prepared with berberine hydrochloride was examined against Candida albicans, C . tropicalis, and C . glabrata . A commercial denture cleanser and a trial denture cleanser that exhibited strong antifungal activity were tested for their effects on Candida spp., the color stability of the dental material, and the surface roughness of acrylic resin plates . The results of these tests revealed that the trial denture cleanser removed 64% to 89% of adhered cells from acrylic resin surfaces and had little effect on the other physical properties tested.

Int Immunopharmacol, 2002 Jan, 2(1), 59 - 67
Application of Candida solubilized cell wall beta-glucan in antitumor immunotherapy against P815 mastocytoma in mice; Tokunaka K et al.; This study was designed to evaluate the antitumor activity of CSBG, purified from the cell wall of Candida albicans IFO1385 . First, as an effect of CSBG on P815 mastocytoma, significant prolonged survival and suppression of the tumor growth were observed . Second, the transfer of spleen cells from CSBG-sensitized BALB/c mice to CDF1 mice led to further suppression of tumor growth as well as P815-immunized spleen cells . Third, CSBG enhanced antitumor immunity in gene therapy using B7-1-transfected P815 cells . These results strongly suggest that CSBG enhances the host defense response to tumor due in part to an adjuvant effect.

Microbiology, 2002 Jan, 148(Pt 1), 29 - 40
Reductive iron uptake by Candida albicans: role of copper, iron and the TUP1 regulator; Knight SA et al.; High-affinity iron uptake by a ferrous permease in the opportunistic pathogen Candida albicans is required for virulence . Here this iron uptake system has been characterized by investigating three distinct activities: an externally directed surface ferric reductase, a membrane-associated PPD (p-phenylenediamine) oxidase and a cellular ferrous iron transport activity . Copper was required for the PPD oxidase and ferrous transport activities . In contrast, copper was not required for iron uptake from siderophores . Addition of iron to the growth medium repressed ferric reductase and ferrous transport, indicating homeostatic regulation . To identify the genes involved, orthologous mutants of Saccharomyces cerevisiae were transformed with a genomic library of C . albicans . CFL95, a gene with sequence similarity to ferric reductases, restored reductase activity to the orthologous S . cerevisiae mutant . CaFTR2 and CaFTR1, genes with homology to ferrous permeases, conferred ferrous transport activity to the orthologous S . cerevisiae mutant . However, neither a genomic library nor CaFET99, a multicopper oxidase homologue and candidate gene for the PPD oxidase, complemented the S . cerevisiae mutant, possibly because of problems with targeting or assembly . Transcripts for CFL95, CaFTR1 and CaFET99 were strongly repressed by iron, whereas the CaFTR2 transcript was induced by iron . Deletion of the TUP1 regulator perturbed the homeostatic control of reductive iron uptake . Incidentally, iron starvation was noted to induce flavin production and this was misregulated in the absence of TUP1 control . The opposite regulation of two iron permease genes and the role of TUP1 indicate that the process of iron acquisition by C . albicans may be more complex and potentially more adaptable than by S . cerevisiae.

Bone Marrow Transplant, 2001 Nov, 28(9), 873 - 8
Candida glabrata and Candida krusei fungemia after high-risk allogeneic marrow transplantation: no adverse effect of low-dose fluconazole prophylaxis on incidence and outcome; Safdar A et al.; Candidemia is a serious complication in patients following allogeneic blood, marrow, and organ transplantation . Fourteen patients developed nosocomial fungemia among 204 allogeneic marrow transplants performed during 1997-1999 . Incidence of hematogenous candidiasis was 6.8 per 100 allogeneic BMT . All 14 had an indwelling central venous catheter (CVC) and fluconazole (100-200 mg daily) was given prophylactically . In 11 (78.5%) neutropenic patients, duration between agranulocytosis and diagnosis of fungemia was (median, +/- s.d.) 10 +/- 8 days . Candida glabrata (53.3%) was the most common yeast species, followed by C . krusei (33.3%), and C . parapsilosis (13.3%) . Candida albicans was conspicuously absent . Ten patients (71.4%) had primary transplant-related complication (>2 days) including hemolytic uremic syndrome/thrombotic thrombocytopenic purpura (HUS/TTP) (n = 5), severe hemorrhagic cystitis (n = 3), and bacteremia (n = 2) . Seven (50.0%) patients expired and in three (21.4%) deaths were attributed to fungemia . The impact of a primary transplant-related complication on short-term survival in this setting was not significant (P = 0.07) (HUS/TTP (P > 0.5); neutropenia (P > 0.5); GVHD (P = 0.35)) . Removal of CVC did not alter outcome in our group (P > or = 0.5) although in patients with persistent fungemia (>72 h), and those with preceding bacteremia, mortality was significantly higher (P = 0.002) . Conventional prognosticators of poor outcome did not adversely effect short-term survival in our transplant recipients with hematogenous candidiasis . The predominance of C . glabrata and C . krusei breakthrough infections was similar to what is seen with high-dose fluconazole (400 mg) prophylaxis, and no adverse effects of low-dose fluconazole in terms of increased incidence of non-susceptible Candida species was seen.

Rocz Akad Med Bialymst, 2001, 46, 326 - 33
Hamycin therapy of murine disseminated candidiasis: efficacy and interaction with fluconazole; Dhuley JN; The efficacy of antifungal agent, hamycin, in a murine model of disseminated candidiasis was studied . Mice were intravenously infected with Candida albicans blastoconidia and treated for 14 days with intraperitoneal hamycin, oral fluconazole, or a combination of these two . Hamycin alone was most efficacious in prolonging survival and in decreasing renal colony counts, usually with complete sterilization of the kidneys by the end of the treatment period . Fluconazole improved survival rates and effected a decrease in renal colony counts, but kidneys were not microbiologically sterilized . Combination therapy with hamycin and fluconazole did not result in a decrease in the efficacy of hamycin by either end point (survival or renal colony counts) . High-pressure liquid chromatographic analysis of Hamycin concentrations in serum indicated lowlevels of absorption of the drug . From the results of the present study, it can be concluded that hamycin is effective in the treatment of murine invasive candidiasis and that the theoretical concern about adverse interactions between the two drug does not apply to the dosages studied in these experiments.

Chin Med J (Engl), 2001 Mar, 114(3), 294 - 6
Epidemiological study of pathogenic fungi in China: 1986 and 1996; Wu S et al.; OBJECTIVE: An annual decade epidemiological survey of pathogenic fungi of inpatients or outpatients includes more than 25 provinces in China has been done in 1986 and 1996 . METHODS: In 1986, there were a total of 9096 strains of pathogenic fungi collected from more than 41 units of 25 provinces in China . 10 years late, 18,085 strains of pathogenic fungi from 41 units of 25 provinces were collected from January 1st to December 31st in 1996 . RESULTS: The results showed that during this decade the prominent pathogenic fungus was Trichophyton rubrum, but its ratio gradually decreased . On the contrary, Candida albicans gradually increased in its ratio from 5th in 1986 to 2nd in 1996 . CONCLUSION: The pathogenic fungi in China have changed greatly in the past decade from 1986 to 1996.

Ther Apher, 2001 Oct, 5(5), 423 - 32
Use of human preconditioned phagocytes for extracorporeal immune support: introduction of a concept; Mitzner SR et al.; Neutrophils are critical effector cells in humoral and innate immunity and play a vital role in phagocytosis and bacterial killing . If they and/or their specific functions are lacking, then immunoparalysis may occur, and severe diseases like systemic inflammatory response syndrome (SIRS) or sepsis can take a fatal course . In this paper, we discuss the possibility of using preconditioned cells in an extracorporeal biohybrid immune support system . A human promyelocytic cell line was stimulated for different times with all-trans retinoic acid . The resulting cells displayed major signs and functions of mature neutrophilic granulocytes including oxygen radical production, phagocytosis of living and dead Escherichia coli, Staphylococcus aureus, Candida albicans, intracellular killing, and interleukin production . The cells can be expanded to yield a sufficient cell mass, and subsequent prestimulation results in an expression of specific neutrophil functions . Extracorporeal bioreactor experiments seem to be feasible to test the benefit in immunoparalysis-associated diseases like SIRS or sepsis.

Eur J Pharm Biopharm, 2002 Jan, 53(1), 115 - 23
A new lipid emulsion formulation with high antimicrobial efficacy using chitosan; Jumaa M et al.; The antimicrobial activity of chitosan in lipid emulsions as well as in aqueous solutions was investigated . Two types of long-chained chitosan were used differing in the molecular weights, degree of the deacetylation and their viscosity: type I, mol . weight 8.7 x 10(4) g/mol, 92% degree of deacetylation and a viscosity of 14 mPa s, type II, mol . weight of 5.32 x 10(5) g/mol, 73% degree of deacetylation and a viscosity of 461 mPa s . In order to assess the pH optimum of the antimicrobial activity of the biopolymer, suspensions of the microorganisms Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans and Aspergillus niger were incubated at different pH-values in lactic acid solution (1% w/v) containing different concentrations of chitosan up to 1.5% (w/v) . Emulsion formulations containing either 0.25%, 0.5% or no chitosan, respectively, were inoculated with the same microorganisms and were incubated at 25 degrees C . The aqueous solutions as well as the emulsions were examined for microbial counts on agar plates after different periods of incubation . After 24 h of incubation in aqueous solutions only the cfu numbers of the bacteria were reduced . Both types of chitosan revealed a pH optimum of their antibacterial activity at pH 5.0-5.1 for P . aeruginosa, and at pH 5.3 for S . aureus . In addition, chitosan with a mol . weight of 8.7 x 10(4) g/mol, high degree of deacetylation and low viscosity showed a higher antimicrobial activity than the other chitosan type of this study . It was found that lipid emulsions containing 0.5% chitosan (type I) conformed to the requirements of the preservation efficacy test for topical formulations according to the European Pharmacopoeia while the emulsion without chitosan and a lactic acid solution with and without the biopolymer did not conform . In hemolysis studies on human erythrocytes, the hemolytic activity of the lipid emulsions with chitosan was assessed . These emulsions showed a negligible hemolytic behavior . The results indicate a use of chitosan as antimicrobial preservative in emulsion formulations for mucosal as well for parenteral applications.

J Microbiol Methods, 2002 Mar, 49(1), 55 - 62
Phagocytosis and intracellular killing of Candida albicans blastoconidia by neutrophils and macrophages: a comparison of different microbiological test systems; Vonk AG et al.; Polymorphonuclear neutrophils (PMN) and mononuclear phagocytes represent an important first line and effector function in the control of Candida infections . Their relative contribution to host defence is frequently assessed by means of microbiological assays . However, reported results are divergent and might well be associated with study design-related issues . In the present study, we compared frequently used microbiological candidacidal assays, with the purpose of determining the most adequate method for assessment of phagocytosis and intracellular killing . We concluded that microbiological assays using yeast-phagocyte suspensions are inappropriate for the assessment of intracellular killing of Candida blastoconidia by murine macrophages, due to adherence or clumping of cells . In contrast, an adherent monolayer of phagocytes can be applied as a single microbiological assay to independently study the process of phagocytosis and intracellular killing, by exudate peritoneal macrophages as well as exudate peritoneal PMN.

Exp Gerontol, 2002 Jan-Mar, 37(2-3), 421 - 6
Melatonin and aging: in vitro effect of young and mature ring dove physiological concentrations of melatonin on the phagocytic function of heterophils from old ring dove; Terron MP et al.; We have studied the circadian rhythm of melatonin in the ring dove (Streptopelia risoria) for different age groups: young (1-1.5 years), mature (3-4 years) and old animals (>8 years) . Melatonin levels were determined by radioimmunoassay . Results showed a significant decline in plasma melatonin levels in old animals when compared with the concentrations observed in the other two age groups, in which maximum (nocturnal) concentrations were 300 pg/ml and minimum (diurnal) concentrations were 50 pg/ml . We analyzed the in vitro effect of the physiological concentrations found in young and mature animals on the heterophils obtained from old animals, evaluating the capacity for ingestion and destruction of Candida albicans, and the oxidative metabolism associate to phagocytosis by determining the superoxide anion levels . Melatonin induced an increase in both the phagocytosis index and the candidicide capacity . This effect was dose-dependent . In relation with the oxidative metabolism, a decline in superoxide anion levels after incubation with both concentrations of the hormone was observed . Thus our results corroborate in this avian species the decline in plasma melatonin levels with advanced age, as well as the enhancing effect of physiological concentrations of melatonin on the phagocytic function.

Mycoses, 2001 Nov, 44(9-10), 422 - 5
Case report . Successful treatment of two cases of post-surgical sternal osteomyelitis, due to Candida krusei and Candida albicans, respectively, with high doses of triazoles (fluconazole, itraconazole); Petrikkos G et al.; Two female patients, aged 75 and 59 years, respectively, with candidal sternal osteomyelitis were successfully treated by the administration of triazoles . Both had developed post-operative wound infection after sternotomy for coronary artery by-pass grafting . Sternal osteomyelitis was confirmed by bone scans with technetium 99Tc and gallium 67Ga . The cultures, from the pus draining at the site of the sternotomy scar, grew Candida krusei in the first case . The fistula closed after a 9-week course of itraconazole therapy (800 mg daily, followed by 600 mg daily) and the patient completed a 6-month period of therapy . The second patient had underlying diabetes mellitus . Post-operatively she developed two fistulae draining pus on the sternum . The pus cultures grew C . albicans . Initial treatment with oral fluconazole (400 mg daily) failed . Subsequent treatment with liposomal amphotericin B also failed . A dramatic improvement was noted when the patient received high doses of fluconazole (800 mg daily) . The fistulae gradually closed after 1 month . Oral fluconazole was continued for 6 months . The cure was confirmed by bone scans . Three years later, both patients remained well.

Mycoses, 2001 Nov, 44(9-10), 368 - 74
Extrusion of fluorescein diacetate by multidrug-resistant Candida albicans; Yang HC et al.; Over-expression of multidrug efflux transporters causes Candida albicans cells to be resistant to azole antifungal agents . There are several kinds of indicator for multidrug resistance (MDR) phenotype of higher eukaryotic cells . Calcein AM is a prefluorochrome that is known as a substrate for multidrug efflux transporters of mammalian cells . We investigated whether calcein AM was also extruded by the ATP-dependent multidrug transporter (cdr1p) of C . albicans . There was no significant difference in the accumulation of calcein AM between MDR cells and drug-susceptible cells of C . albicans even with sodium azide, suggesting that calcein AM may not be associated with the CDR1-gene-related multidrug efflux system of C . albicans . However, a structurally related prefluorochrome derivative, fluorescein diacetate (FDA), was shown to be extruded by the CDR1 mRNA-overexpressing yeast cells . In comparison with drug-susceptible cells, the resistant cells emitted very weak fluorescence when stained with FDA . Furthermore sodium azide increased the fluorescence of the resistant cells more than 20 times, whereas the fluorescence in the drug-susceptible cells with FDA and sodium azide was three to four times stronger . These results suggested that FDA might be extruded by the CDR1-related multidrug efflux transporter of C . albicans.

Mycoses, 2001 Nov, 44(9-10), 361 - 7
Phospholipases of Candida albicans; Niewerth M et al.; Infections due to Candida albicans are frequent and of clinical importance . Especially at a time of increasing organ transplantations, HIV infections, and resistance to antimicrobial agents a profound knowledge of the interaction between C . albicans and host tissue is mandatory . In addition to secreted aspartyl proteinase, dimorphism, cell surface composition, and toxin production phospholipases are a main factor in pathogenicity . Up to the present, many different groups and subgroups of phospholipases have been detected . These different enzymes are related to various types of aggressive and defensive actions . These range from active invasion of host cell tissue to growth control and remodelling of the yeast cell membrane . It is clear that a multiplicity of factors must co-operate to overcome the host's defences . Yet it can be supposed today that phospholipases are one important factor in this complex interaction . Therefore the known phospholipases of C . albicans are described in detail under clinical aspects.

J Med Microbiol, 2001 Dec, 50(12), 1105 - 10
Comparative performance of Fungichrom I, Candifast and API 20C Aux systems in the identification of clinically significant yeasts; Gundes SG et al.; To compare the performance of current chromogenic yeast identification methods, three commercial systems (API 20C Aux, Fungichrom I and Candifast) were evaluated in parallel, along with conventional tests to identify yeasts commonly isolated in this clinical microbiology laboratory . In all, 116 clinical isolates, (68 Candida albicans, 12 C . parapsilosis, 12 C . glabrata and 24 other yeasts) were tested . Germ-tube production, microscopical morphology and other conventional methods were used as standards to definitively identify yeast isolates . The percentage of isolates identified correctly varied between 82.7% and 95.6% . Overall, the performance obtained with Fungichrom I was highest with 95.6% identification (111 of 116 isolates) . The performance of API 20C Aux was higher with 87% (101 of 116 isolates) than that of Candifast with 82.7% (96 of 116) . The Fungichrom I method was found to be rapid, as 90% of strains were identified after incubation for 24 h at 30 degrees C . Both of the chromogenic yeast identification systems provided a simple, accurate alternative to API 20C Aux and conventional assimilation methods for the rapid identification of most commonly encountered isolates of Candida spp . Fungichrom seemed to be the most appropriate system for use in a clinical microbiology laboratory, due to its good performance with regard to sensitivity, ease of use and reading, rapidity and the cost per test.

Arch Pharm (Weinheim), 2001 Oct, 334(10), 305 - 8
Synthesis and antifungal activity of 3,3'-ethylenebis(5-alkyl-1,3,5-thiadiazine-2-thiones); Hussein MA et al.; In a search for promising antifungal compounds, nine new 3,3'-ethylenebis(5-alkyl-1,3,5-thiadiazine-2-thiones) were synthesized by the reaction of ethylene diamine, carbon disulfide, formaldehyde, and the appropriate alkyl amine . The title compounds were tested for their antifungal activity in vitro against pathogenic (Trichophyton rubrum and Candida albicans), phytopathogenic (Penicillum expansum, Trichoderma hazianum, and Fasarium oxysporum), and aflatoxin producing (Aspergillus flavus) fungi . These compounds exhibited varied inhibitory effects on growth or sporulation of some tested fungal species.

J Hosp Infect, 2001 Aug, 48 Suppl A, S80 - 3
How to improve instrument disinfection by ultrasound; Jatzwauk L et al.; Ultrasound technologies have a wide range of hospital and dental applications which include cleaning and disinfection of surgical and dental instruments . We measured the germicidal efficacy of sonication, with or without chemical disinfectants, in an ultrasonic bath delivering a frequency of 35 kHz and an intensity of 0.66 W/cm2 . Cultures of Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans were exposed to ultrasound and to an amine-based disinfectant in non-bactericidal concentrations . Ultrasonication for 60 min alone did not cause a significant killing of the bacteria and yeast . However, we were able to show that sonication can act as a powerful synergistic agent to increase the cidal efficacy of the disinfectant against S . aureus and P . aeruginosa . C . albicans was more resistant to the combination of ultrasound and chemical disinfection . The key role in the action of ultrasound in cleaning of instruments and perhaps in enhanced disinfection is played by cavitation phenomena . The distribution of cavitations in an ultrasonic bath is not homogenous . We found a similar synergistic effect of ultrasound and disinfectant in positions with low cavitation . The synergistic effect was not reduced inside rubber tubes . Before ultrasound can be accepted as an integral part of the cleaning and disinfection process of medical instruments, the influence of intensity and frequency of sonication and the effects of cavitation must be clarified.

Oncology (Huntingt), 2001 Nov, 15(11 Suppl 9), 11 - 4
Overview of systemic fungal infections; Rolston K; A steady increase in the frequency of invasive fungal infections has been observed in the past 2 decades, particularly in immunosuppressed patients . In recipients of bone marrow transplants, Candida albicans and Aspergillus fumigatus remain the primary pathogens . In many centers, however, Candida species other than C albicans now predominate, and many cases of aspergillosis are due to species other than A fumigatus . Additionally, heretofore unrecognized and/or uncommon fungal pathogens are beginning to emerge, including Blastoschizomyces capitatus, Fusarium species, Malassezia furfur, and Trichosporon beigelii . These opportunistic fungal pathogens are associated with various localized and disseminated clinical syndromes, and with substantial morbidity and mortality . These established, invasive mycoses, particularly in bone marrow transplant recipients, are the focus of this discussion.

Eur J Med Chem, 2001 Nov-Dec, 36(11-12), 887 - 97
Synthesis and biological activities of indolocarbazoles bearing amino acid residues; Moreau P et al.; Three indolocarbazole compounds bearing a tripeptide or a lysine group attached to one of the indole nitrogens via a propylamino chain and two rebeccamycin derivatives bearing a lysine residue on the sugar moiety were synthesised with the aim of improving the binding to DNA and the antiproliferative activities . Four tumour cell lines, from murine L1210 leukemia, human HT29 colon carcinoma, A549 non-small cell lung carcinoma and K-562 leukemia, were used to evaluate the cytotoxicity of the drugs . Their effects on the cell cycle of L1210 cells and their antimicrobial properties against two Gram-positive bacteria Bacillus cereus and Streptomyces chartreusis, a Gram-negative bacterium Escherichia coli and a yeast Candida albicans were also investigated.

FEMS Immunol Med Microbiol, 2001 Dec, 32(1), 27 - 32
A clinical study on the association of Trichomonas vaginalis and Mycoplasma hominis infections in women attending a sexually transmitted disease (STD) outpatient clinic; van Belkum A et al.; Swabs from the posterior vaginal fornix were obtained from 804 consecutive female patients visiting a large Dutch sexually transmitted diseases (STD) outpatient clinic . A detailed clinical history was obtained and complaints concerning the lower genital tract, such as vaginal discharge or vulval and vaginal irritation, were recorded . Patients were examined and the presence of non-physiological vaginal secretions was established by speculum examination . The swabs were monitored for bacterial vaginosis (BV) or Candida albicans infection . PCR diagnosis of Chlamydia trachomatis and Trichomonas vaginalis was performed as well . Four groups of patients (n=14-21) with BV or single infections caused by one of these three pathogens and a control group with no pathogens were selected and Mycoplasma hominis PCR was performed additionally . At clinical presentation, controls and single-infected patient groups were comparable with regard to complaints of the lower genital tract and sexual risk behavior defined as having prior STDs and/or admitted prostitution . Only in the T . vaginalis-positive group significantly more women reporting sexual risk behavior were found than in controls . In agreement with former in vitro observations, an in vivo association between the PCR-detected presence of M . hominis and T . vaginalis was established . In 79% of all samples positive for T . vaginalis, M . hominis could be detected, as compared to only 6% in control samples (P=0.0004) . However, since single infections by either of the two pathogens were regularly observed, there does not seem to be an exclusive association between the species, as the bacterium is also more frequently found in cases of BV (P=0.026) . Co-infection of M . hominis with C . albicans (11%) or C . trachomatis (0%) did not differ significantly from controls (6%) . M . hominis did not associate with complaints of the lower genital tract . However, if all groups were combined there appears to be a very significant association between the presence of M . hominis and sexual risk behavior (P=0.0004) . M . hominis and sexual risk behavior were more closely associated than M . hominis and T . vaginalis . No indications were found for an enhanced pathogenicity by either of the symbionts.

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