Candida albicans

Diagn Microbiol Infect Dis, 2001 Nov, 41(3), 113 - 9
Variation in random amplified polymorphic DNA (RAPD) profiles specific to fluconazole-resistant and -sensitive strains of Candida albicans; Jain P et al.; Random amplified polymorphic DNA analysis was used to detect genotype relatedness among clinical fluconazole-resistant and -sensitive strains of Candida albicans recovered from twenty HIV-infected patients having oropharyngeal candidiasis . Sensitive strains were obtained from a local hospital and were from patients that had not been treated with azole drugs while resistant strains were recovered from patients in different parts of Europe and their resistance was a consequence of drug-treatment given to the patients . On amplification with different arbitrary sequence decamer primers, the results demonstrated a homogeneous banding pattern for all sensitive strains that was distinct from that obtained in case of the resistant strains . The DNA profiles of strains were thus broadly clustered into two major groups of resistant and sensitive strains . The RAPD technique may be useful in differentiating fluconazole-resistant strains from the -sensitive ones for early identification of resistant isolates from AIDS patients.

J Antimicrob Chemother, 2001 Dec, 48(6), 781 - 6
Bumetanide annihilation of amphotericin B-induced apoptosis and cytotoxicity is due to its effect on cellular K+ flux; Marklund L et al.; The antifungal antibiotic amphotericin B causes considerable toxic effects during clinical therapy . We have shown previously that amphotericin B-induced cytotoxicity and apoptosis were eradicated by the Na+, K+, 2Cl- cotransport inhibitor bumetanide . To elucidate the role of K+ flux and the activity of Na+, K+ ATPase and Na+, K+, 2Cl- cotransport in apoptosis and cytotoxicity induced by amphotericin B alone and combined with bumetanide, we quantified the influx and efflux of K+ of mesothelioma cells (P31) using the K+ analogue 86Rb+ with ouabain (100 micromol/L) as the K+ influx probe . To determine the susceptibility of Candida albicans to amphotericin B when combined with bumetanide we used a plate diffusion method . Amphotericin B or bumetanide alone significantly stimulated 86Rb+ efflux during the first 15 min . However, when added simultaneously, the cellular 86Rb+ efflux was markedly decreased . Amphotericin B (3 mg/L) had no effect on immediate (15 min) total 86Rb+ influx . When bumetanide (100 micromol/L) was added, the total 86Rb+ influx was markedly reduced due to inhibition of augmented Na+, K+, 2Cl- cotransport and low Na+, K+ ATPase activity . Bumetanide did not affect the susceptibility of C . albicans to amphotericin B, which suggests that bumetanide or related drugs could be used in antifungal therapy to increase amphotericin B effectiveness without increasing its adverse effects . We suggest that bumetanide hampering of amphotericin B-induced cytotoxicity and apoptosis could be due to an immediate reduction of cellular K+ efflux as well as disordered K+ influx.

Enferm Infecc Microbiol Clin, 2001 Aug-Sep, 19(7), 304 - 7
{Nosocomial fungemias in a general hospital . Epidemiology and prognostic factors . Prospective study 1993-1998}; Gomez J et al.; BACKGROUND: Nosocomial fungemias are infections with a high mortality rate . In last years the incidence of these infections has increased probably because of the growing population of immunocompromised patients who undergo aggressive diagnostic and therapeutic techniques . OBJECTIVE: To know the epidemiologic characteristics, risk factors, clinical features and prognosis of fungemia . PATIENTS AND METHODS: We prospectively evaluated all the patients with proven fungemia in our center during a 5 year-period . After finishing antifungal treatment a minimum follow-up of 1 month was carried out . Fungal isolation and identification were performed by standard tests . RESULTS: During the period of study we evaluated 81 patients with an episode of nosocomial fungemia . Global incidence was 0,9 episodes per thousand admitted patients . Candida albicans was the more frequently isolated species (n=53), followed by C . parapsilosis (n=11), C . tropicalis (n=6) and C . glabrata (n=5) . Most of the patients had a central intravenous line and were on parenteral nutrition therapy . All of them previously received at least one course of broad-spectrum antibiotics . Overall mortality was 49,6% . A worst prognosis was significantly associated with: age over 65 years, surgical procedures during present admission, leucocytosis, shock, and delay in antifungal treatment . CONCLUSIONS: Fungal bloodstream infection incidence is high in our environment . It is associated with a high mortality rate, specially in patients in whom the beginning of antifungal treatment was delayed . A higher clinical suspicion index may improve the poor outcome in these patients.

Yeast, 2001 Nov, 18(15), 1391 - 6
Aquaporin in Candida: characterization of a functional water channel protein; Carbrey JM et al.; The Candida albicans genome database contains one ORF with homology to aquaporins, AQY1 . Xenopus oocytes injected with cRNA encoding C . albicans Aqy1p displayed a coefficient of water permeability (P(f)) that was equivalent to the P(f) for oocytes injected with the cRNA of S . cerevisiae Aqy1p . In addition, as seen in Saccharomyces for Aqy1p and Aqy2p, deletion of AQY1 in C . albicans resulted in cells that were less sensitive than wild-type to osmotic shock . In Saccharomyces, aquaporin null cells also have a cell surface that is more hydrophobic . However, unlike Saccharomyces, there was no effect on the cell surface hydrophobicity, flocculation or cell aggregation in aqy1 null C . albicans cells . Perhaps as a result, there was no difference between the virulence of C . albicans wild-type and aqy1 null strains in a murine model for systemic candidiasis .

Yeast, 2001 Nov, 18(15), 1383 - 9
Ynl038wp (Gpi15p) is the Saccharomyces cerevisiae homologue of human Pig-Hp and participates in the first step in glycosylphosphatidylinositol assembly; Yan BC et al.; Glycosylphosphatidylinositols (GPIs) are found in all eukaryotes and are synthesized in a pathway that starts with the transfer of N-acetylglucosamine (GlcNAc) from UDP-GlcNAc to phosphatidylinositol (PI) . This reaction is carried out by a protein complex, three of whose subunits in humans, hGpi1p, Pig-Cp and Pig-Ap, have sequence and functional homologues in the Saccharomyces cerevisiae Gpi1, Gpi2 and Gpi3 proteins, respectively . Human GlcNAc-PI synthase contains two further subunits, Pig-Hp and PigPp . We report that the essential YNL038w gene encodes the S . cerevisiae homologue of Pig-Hp . Haploid YNL038w-deletion strains were created, in which Ynl038wp could be depleted by repressing YNL038w expression using the GAL10 promoter . Depletion of Ynl038wp from membranes virtually abolished in vitro GlcNAc-PI synthetic activity, indicating that Ynl038wp is necessary for GlcNAc-PI synthesis in vitro . Further, depletion of Ynl038wp in an smp3 mutant background prevented the formation of the trimannosylated GPI intermediates that normally accumulate in this late-stage GPI assembly mutant . Ynl038wp is therefore required for GPI synthesis in vivo . Because YNL038w encodes a protein involved in GPI biosynthesis, we designate the gene GPI15 . Potential Pig-Hp/Gpi15p counterparts are also encoded in the genomes of Schizosacchomyces pombe and Candida albicans .

Cell Biochem Funct, 2001 Dec, 19(4), 229 - 35
3-Nitrocoumarin is an efficient inhibitor of budding yeast phospholipase-C; Tisi R et al.; 3-Nitrocoumarin is described in the literature as a specific inhibitor of mammalian phospholipase-C and here we studied the effect of 3-nitrocoumarin on budding yeast phosphatidylinositol-specific phospholipase-C and its effect on yeast growth . 3-Nitrocoumarin is a powerful inhibitor in vitro of the yeast Plc1 protein with an IC(50) of 57 nM and it is also an inhibitor of yeast growth in minimal media at comparable concentrations . Moreover at the same concentration it inhibits the glucose-induced PI-turnover . Since the effects of 3-nitrocoumarin on yeast growth are superimposable on the growth phenotype caused by PLC1 gene deletion we can conclude that 3-nitrocoumarin is a specific and selective inhibitor of yeast phospholipase-C . In addition we show that 3-nitrocoumarin was also an effective inhibitor of the pathogenic yeast Candida albicans .

Minerva Stomatol, 2001 Nov-Dec, 50(11-12), 345 - 9
Phagocytosis and killing of Candida albicans of polymorphonuclear cells in patients with organ transplant of periodontal disease; Maccarinelli G et al.; BACKGROUND: The easiest defence system carried out by the organism, the inflammatory response, happens with the support of phagocyting cells: the polymorphonuclear leukocytes (PMNL) or neutrophils are the most important cell line acting as the first defence of the organism against bacterial agents . Previous studies have shown a correlation between a reduction of the immune function and development of periodontal disease . Furthermore, it is well known that transplant patients show a variety of oral lesions as a consequence of their therapy, in particular to immunosuppressive drugs . The aim of this study is to evaluate the phagocytosis and killing functions of PMNL in transplant patients and in patients with periodontal disease in comparison with a group of healthy subjects . METHODS: PMNL, were isolated by spontaneous sedimentation from heparinized blood and centrifugation of plasma on density medium . Phagocytosis rate was expressed as the percentage of Candida albicans phagocyted after 20' incubation and phagocyting PMNLs . Intracellular killing was expressed as the percentage of yeast cells killed . RESULTS: We did not find a significant decrease of phagocytosis in transplant patients and patients with periodontal disease while these two groups of patients showed a decrease of PMNL killing activity in respect to healthy controls, an effect which was unrelated to the severity of periodontal disease . CONCLUSIONS: These results suggest that a reduction of killing activity, either spontaneous or drug-induced, would contribute to the development of periodontal disease.

J Ethnopharmacol, 2002 Jan, 79(1), 43 - 52
Yucatec Mayan medicinal plants: evaluation based on indigenous uses; Ankli A et al.; As part of an ethnopharmacological field study 48 medicinal plants were evaluated using several biological assays with the goal to obtain information on the pharmacological effects of these plants, which may be of direct relevance to the indigenous uses . Three species used to treat gastrointestinal disorders showed remarkable activity against Helicobacter pylori . One of them showed activity against Giardia duodenalis . Cytotoxic effects against KB cells were found for six species . In the group of plants used for dermatological conditions several species were active against gram-positive bacteria and Candida albicans . Two plant species of this group were found to be active in an Nuclear Factor-kappaB (NF-kappaB) assay measuring inhibition of this pro-inflammatory transcription factor . A species of the Solanaceae, applied in cases of pain and fever, showed a weak activity against Plasmodium falciparum . One species traditionally used for diabetes exhibited antihyperglycemic activity . None of the six species from the group of 'women's medicine' showed relevant affinity to the D(2) dopamine receptor . Based on this evaluation, plants with strong activities should be further investigated phytochemically and pharmacologically to identify active fractions and compounds.

J Inorg Biochem, 2001 Dec 15, 87(4), 227 - 35
Substrate recognition sites in 14alpha-sterol demethylase from comparative analysis of amino acid sequences and X-ray structure of Mycobacterium tuberculosis CYP51; Podust LM et al.; The crystal structure of 14alpha-sterol demethylase from Mycobacterium tuberculosis (MTCYP51) {Proc . Natl . Acad . Sci . USA 98 (2001) 3068-3073} provides a template for analysis of eukaryotic orthologs which constitute the CYP51 family of cytochrome P450 proteins . Putative substrate recognition sites (SRSs) were identified in MTCYP51 based on the X-ray structures and have been compared with SRSs predicted based on Gotoh's analysis {J . Biol . Chem . 267 (1992) 83-90} . While Gotoh's SRS-4, 5, and 6 contribute in formation of the putative MTCYP51 substrate binding site, SRS-2 and 3 likely do not exist in MTCYP51 . SRS-1, as part of the open BC loop, in the conformation found in the crystal can provide only limited contacts with the sterol . However, its role in substrate binding might dramatically increase if the loop closes in response to substrate binding . Thus, while the notion of SRSs has been very useful in leading to our current understanding of P450 structure and function, their identification by sequence alignment between distant P450 families will not necessarily be a good predictor of residues associated with substrate binding . Localization of CYP51 mutation hotspots in Candida albicans azole resistant isolates was analyzed with respect to SRSs . These mutations are found to be outside of the putative substrate interacting sites indicating the preservation of the protein active site under the pressure of azole treatment . Since the mutations residing outside the putative CYP51 active side can profoundly influence ligand binding within the active site, perhaps they provide insight into the basis of evolutionary changes which have occurred leading to different P450s.

J Bacteriol, 2002 Jan, 184(1), 29 - 42
Molecular and phenotypic analysis of CaVRG4, encoding an essential Golgi apparatus GDP-mannose transporter; Nishikawa A et al.; Cell surface mannan is implicated in almost every aspect of pathogenicity of Candida albicans . In Saccharomyces cerevisiae, the Vrg4 protein acts as a master regulator of mannan synthesis through its role in substrate provision . The substrate for mannosylation of proteins and lipids in the Golgi apparatus is GDP-mannose, whose lumenal transport is catalyzed by Vrg4p . This nucleotide sugar is synthesized in the cytoplasm by pathways that are highly conserved in all eukaryotes, but its lumenal transport (and hence Golgi apparatus-specific mannosylation) is a fungus-specific process . To begin to study the role of Golgi mannosylation in C . albicans, we isolated the CaVRG4 gene and analyzed the effects of loss of its function . CaVRG4 encodes a functional homologue of the S . cerevisiae GDP-mannose transporter . CaVrg4p localized to punctate spots within the cytoplasm of C . albicans in a pattern reminiscent of localization of Vrg4p in the Golgi apparatus in S . cerevisiae . Like partial loss of ScVRG4 function, partial loss of CaVRG4 function resulted in mannosylation defects, which in turn led to a number of cell wall-associated phenotypes . While heterozygotes displayed no growth phenotypes, a hemizygous strain, containing a single copy of CaVRG4 under control of the methionine-repressible MET3 promoter, did not grow in the presence of methionine and cysteine, demonstrating that CaVRG4 is essential for viability . Mutant Candida vrg4 strains were defective in hyphal formation but exhibited a constitutive polarized mode of pseudohyphal growth . Because the VRG4 gene is essential for yeast viability but does not have a mammalian homologue, it is a particularly attractive target for development of antifungal therapies.

Pediatr Infect Dis J, 2001 Dec, 20(12), 1119 - 24
Risk factors for Candida species colonization of neonatal intensive care unit patients; Saiman L et al.; BACKGROUND: Candida spp . are increasingly important pathogens in neonatal intensive care units (NICU) . Prior colonization is a major risk factor for candidemia, but few studies have focused on risk factors for colonization, particularly in NICU patients . METHODS: A prospective, multicenter cohort study was performed in six NICUs to determine risk factors for Candida colonization . Infant gastrointestinal tracts were cultured on admission and weekly until NICU discharge and health care worker hands were cultured monthly for Candida spp . RESULTS: The prevalence of Candida spp . colonization was 23% (486 of 2157 infants); 299 (14%), 151 (7%) and 74 (3%) were colonized with Candida albicans, Candida parapsilosis and other Candida spp., respectively . Multiple logistic regression analysis adjusting for length of stay, birth weight < or = 1000 g and gestational age < 32 weeks revealed that use of third generation cephalosporins was associated with either C . albicans (155 incident cases) or C . parapsilosis (104 incident cases) colonization . Use of central venous catheters or intravenous lipids were risk factors for C . albicans, whereas delivery by cesarean section was protective . Use of H2 blockers was an independent risk factor for C . parapsilosis . Of 2989 cultures from health care workers' hands, 150 (5%) were positive for C . albicans and 575 (19%) for C . parapsilosis, but carriage rates did not correlate with NICU site-specific rates for infant colonization . CONCLUSIONS: We speculate that NICU patients acquire Candida spp., particularly C . parapsilosis, from the hands of health care workers . H2 blockers, third generation cephalosporins and delayed enteral feedings alter gastrointestinal tract ecology, thereby facilitating colonization.

Microbiology, 2001 Dec, 147(Pt 12), 3323 - 34
Genetically engineered human salivary histatin genes are functional in Candida albicans: development of a new system for studying histatin candidacidal activity; Baev D et al.; Histatins are a structurally related family of salivary proteins known as histidine-rich proteins that are produced and secreted by the human major salivary glands . In vitro, histatins are potent cytotoxic proteins with selectivity for pathogenic yeasts including Candida albicans . Studies that investigate the mechanism of action of histatin proteins upon this important human pathogen have used a candidacidal assay in which the histatin is applied extracellularly . In order to develop a model system to study the mechanism of histatin action independently from binding and translocation events, the authors constructed C . albicans strains that contain chromosomally encoded human salivary histatin genes under the control of a regulated promoter . Intracellular expression of either histatin 5 or histatin 3 induced cell killing and ATP release in parallel . Since histatin killing can be initiated solely from intracellular sites, extracellular binding and internalization are preceding transport events . Thus the mechanism of histatin-induced ATP release does not require extracellular binding, and intracellular targets alone can activate ATP release . By employing a codon-optimization strategy it was shown that expression of heterologous sequences in C . albicans can be a useful tool for functional studies.

Oral Microbiol Immunol, 2001 Dec, 16(6), 383 - 5
Prevalence of yeast among children in Nigeria and the United States; Jabra-Rizk MA et al.; Fungal infections have gained considerable importance over the last decade as a result of significant increase in the incidence of opportunistic and systemic candidosis . Although Candida albicans is the predominant causative agent of candidosis, particularly oral disease, recently an epidemiological trend has been observed where other less pathogenic species of Candida, including the newly characterized species Candida dubliniensis, are emerging as significant opportunistic pathogens . The present study aimed to screen for the presence of C . dubliniensis and to compare the recovery of yeast species from 30 seemingly healthy and 30 HIV-positive children in the United States, as well as from 64 malnourished Nigerian children . Oral samples were cultured for fungal growth, and all germ tube and chlamydospore positive isolates were tested for ability to grow at 45 degrees C to differentiate between C . albicans and C . dubliniensis . All isolates were speciated based on colony color production on CHROMagar medium and sugar assimilation profiles . Among the 30 HIV-positive children, 15 (50%) were positive for fungus; 12 were positive for C . albicans, with one of the latter also positive for Candida glabrata, and three were found to harbor C . dubliniensis . Among the 30 non-HIV-positive children, five C . albicans and four C . dubliniensis isolates were recovered . No C . dubliniensis isolates were recovered from the Nigerian group . However, eight other different yeast species were recovered from 31 (48.4%) of the 64 Nigerian children sampled, with six of them growing a combination of species . In comparing the data from the Nigerian and United States children, the frequency of yeasts in the malnourished Nigerian group was considerably higher . The most striking difference between the two groups was in the variety of the usually less encountered and less pathogenic yeast species recovered from the Nigerian population . The findings support previously reported observations that there may be intrinsic differences between different populations sampled and that malnutrition might favor the presence of yeast species other than C . albicans.

Oral Microbiol Immunol, 2001 Dec, 16(6), 358 - 63
Irradiation-induced oral candidiasis in an experimental murine model; Farah CS et al.; The aim of this experiment was to establish a mouse model of irradiation-induced oral candidiasis and to explore the cellular populations and mechanisms by which the infection is cleared from the oral mucosa . BALB/c mice received irradiation to the head and neck equivalent to 800 Rad using a Cobalt 60 gamma source . Both irradiated and non-irradiated mice were infected orally with 1 x 10(8) Candida albicans yeasts . Compared with untreated controls, irradiated animals developed a more severe infection of longer duration, with hyphae penetrating the oral mucosa . Monoclonal antibody depletion of CD4+ but not CD8+ T cells from the systemic circulation prolonged the infection in irradiated mice, but not in controls . Supernatants of submandibular and superficial cervical lymph node cultures from irradiated animals demonstrated significantly higher titers of interleukin-12, but similar levels of interferon-gamma compared with controls . Screening for cytokine production by an RNase protection assay detected only macrophage migration inhibition factor in irradiated and non-irradiated oral tissues from day 8 onwards . The results of this study demonstrate a requirement for CD4+ T cells in the recovery from oral candidiasis induced by head and neck irradiation in mice, and are consistent with a role for Th1-type cytokines in host resistance.

Mol Microbiol, 2001 Nov, 42(4), 981 - 93
Transcript profiling in Candida albicans reveals new cellular functions for the transcriptional repressors CaTup1, CaMig1 and CaNrg1; Murad AM et al.; The pathogenic fungus, Candida albicans contains homologues of the transcriptional repressors ScTup1, ScMig1 and ScNrg1 found in budding yeast . In Saccharomyces cerevisiae, ScMig1 targets the ScTup1/ScSsn6 complex to the promoters of glucose repressed genes to repress their transcription . ScNrg1 is thought to act in a similar manner at other promoters . We have examined the roles of their homologues in C . albicans by transcript profiling with an array containing 2002 genes, representing about one quarter of the predicted number of open reading frames (ORFs) in C . albicans . The data revealed that CaNrg1 and CaTup1 regulate a different set of C . albicans genes from CaMig1 and CaTup1 . This is consistent with the idea that CaMig1 and CaNrg1 target the CaTup1 repressor to specific subsets of C . albicans genes . However, CaMig1 and CaNrg1 repress other C . albicans genes in a CaTup1-independent fashion . The targets of CaMig1 and CaNrg1 repression, and phenotypic analyses of nrg1/nrg1 and mig1/mig1 mutants, indicate that these factors play differential roles in the regulation of metabolism, cellular morphogenesis and stress responses . Hence, the data provide important information both about the modes of action of these transcriptional regulators and their cellular roles . The transcript profiling data are available at http://www.pasteur.fr/recherche/unites/RIF/transcriptdata/.

Clin Microbiol Infect, 2001 Nov, 7(11), 641 - 5
Unusual etiology of visual loss in an HIV-infected patient due to endogenous endophthalmitis; Miailhes P et al.; Disseminated candidiasis, especially ocular infections such as endophthalmitis, is uncommon in HIV-infected patients . We report a case of candidal endophthalmitis in an HIV-positive non-drug-user patient, following candidemia from a cutaneous abscess at the site of a peripheral catheter . Ocular disease was revealed by a visual decrease in the left eye . DNA analysis using RAPD showed identical patterns of Candida albicans isolated from the skin and eye . Combination therapy with high-dose fluconazole and intravenous amphotericin B was performed . Two intravitreal amphotericin B injections and a vitrectomy were administered because of an amblyopic right eye and severe vitritis . The outcome was favorable without relapse at 18 months.

Med Hypotheses, 2001 Nov, 57(5), 570 - 2
Does gastrointestinal Candida albicans prevent ubiquinone absorption?
Krone CA, Elmer GW, Ely JT, Fudenberg HH, Thoreson J.
Ubiquinones (coenzyme Qs (CoQ)) are essential for oxidative phosphorylation in yeasts and humans, although the isomers present in each are different . The human coenzyme Q, CoQ10, is administered orally for the treatment of heart disease and other disorders . Some patients, however, require much higher doses than others to attain a therapeutic CoQ10 blood level . We propose that one possible explanation for this variability is Candida colonization of the GI tract . Many common medical treatments including antibiotics and anti-hyperchlorhydric agents increase the risk of GI tract Candida colonization . Subsequent uptake and utilization of supplemental CoQ10 by the yeast could diminish availability for the human subject . Data from one patient and an in vitro pilot study using two pathogenic strains of C . albicans support this hypothesis . If C . albicans in the GI tract can hinder availability and interfere with therapeutic effects of CoQ10, it could be of clinical significance for large numbers of patients .

Kobe J Med Sci, 2001 Aug, 47(4), 161 - 7
Use of chromogenic tube and methyl blue-sabouraud agar for the identification of Candida albicans strains; Yucesoy M et al.; This study was performed to investigate the use of chromogenic tube and methyl blue-Sabouraud agar for the presumptive identification of Candida albicans . 124 clinical isolates, including 111 C.albicans and 13 Candida spp strains, which had been identified by morphology on cornmeal tween 80 agar and Vitek automated identification system, were included . Three different identification procedures, a) germ tube test, b) chromogenic tube test by using CHROMagar Candida and c) methyl blue-Sabouraud agar test, were performed to the strains . 88 of 111 (79.3%) C.albicans strains were detected to be positive by germ tube test . 87 (78.4%), 97 (87.4%) and 102 (91.9%) of these isolates were identified as C.albicans by chromogenic tube test after 2, 8 and 24 hours of incubation, respectively . 88 (79.3%), 92 (82.9%) and 88 (79.3%) of the isolates were correctly identified as C.albicans by methyl blue-Sabouraud agar test after 2, 8 and 24 hours of incubation, respectively . The sensitivity and specificity values were found to be 79.3 and 69.2 for the germ tube test . These values ranged between 78.4-91.9% and 69.2-76.9% for chromogenic tube test and 79.3-82.9% and 76.9-84.6% for methyl blue-Sabouraud agar depending on the incubation period . It can be concluded that the use of chromogenic tube and methyl blue-Sabouraud agar are rapid, simple and objective methods for the identification of C.albicans strains.

Planta Med, 2001 Nov, 67(8), 771 - 3
Essential oils from Piper cernuum and Piper regnellii: antimicrobial activities and analysis by GC/MS and 13C-NMR; Costantin MB et al.; The essential oils of Piper cernuum and Piper regnellii leaves were analyzed by gas chromatography-mass spectrometry (GC-MS) and the results were compared to that obtained by means of a program designed to analyse (13)C-NMR data of complex mixtures . Bicyclogermacrene (21.88 %)/beta-caryophyllene (20.69 %) and myrcene (52.60 %)/linalool (15.89 %) were the major constituents in essential oil from leaves of P . cernuum and P . regnellii, respectively . Both essential oils presented growth inhibitory activities against Staphylococcus aureus and Candida albicans.

Curr Opin Microbiol, 2001 Dec, 4(6), 728 - 35
Transcriptional control of dimorphism in Candida albicans; Liu H; Candida albicans uses a network of multiple signaling pathways to control the yeast-->hypha transition . These include a mitogen-activated protein kinase pathway through Cph1, the cAMP-dependent protein kinase pathway via Efg1, a pH-responsive pathway through Rim101, the Tup1-mediated repression through Rfg1 and Nrg1, and pathways represented by transcription factors Cph2, Tec1 and Czf1 . These pathways control the transcription of a common set of hypha-specific genes, many of which encode known virulence factors . The link between the signaling pathways and hyphal elongation is currently unknown, but there is evidence to suggest that Cdc42 likely plays a key role in hyphal morphogenesis . Unlike pseudohyphal growth in Saccharomyces cerevisiae, hyphal elongation is regulated independently of the cell cycle . Cellular differences between pseudohyphae and hyphae are further revealed by septin localization.

FEMS Microbiol Lett, 2001 Nov 13, 204(2), 323 - 8
Ura-status-dependent adhesion of Candida albicans mutants; Bain JM et al.; Gene disruptions in the diploid opportunistic human fungal pathogen Candida albicans are usually created using multiple rounds of targeted integration called the 'ura-blaster' method . Resulting heterozygous and homozygous null mutants can be auxotrophic (Ura(-)) or prototrophic (Ura(+)) for uracil biosynthesis . Here we demonstrate that the Ura-status of otherwise isogenic mutants affected the adhesion of C . albicans . Moreover the effect of Ura-status on adhesion was also dependent on the null mutant background, the nature of the underlying surface and the carbon source for growth . Therefore the Ura-status is not neutral in determining adhesive properties of C . albicans mutants that are generated via the ura-blaster protocol.

Phytochemistry, 2001 Dec, 58(7), 1141 - 5
Antifungal highly oxygenated guaianolides and other constituents from Ajania fruticulosa; Meng JC et al.; Three highly oxygenated guaianolides were isolated from the aerial parts of Ajania fruticulosa along with 17 known phytochemicals including a triterpene (alpha-amyrin), two plant sterols (beta-sitosterol, daucosterol), four flavonoids (axillarin, centaureidin, santin and 5,7,4'-trihydroxy-3,3'-dimethoxyflavone), and ten sesquiterpenes {1alpha-hydroperoxy-4beta,8alpha,10alpha,13-tetrahydroxyguaia-2-en-12,6alpha-olide, 1alpha-hydroperoxy-4alpha,10alpha-dihydroxyguaia-9alpha-angeloyloxyguaia-2,11(13)-dien-12,6alpha-olide, 3beta,4alpha-dihydroxyguaia-11(13),10(14)-dien-12,6alpha-olide, 1alpha,4alpha,10alpha-trihydroxy-9alpha-angeloyloxyguaia-2,11(13)-dien-12,6alpha-olide, 1beta,2beta-epoxy-3beta,4alpha,10alpha-trihydroxy-guaia-11(13)-en-12,6alpha-olide and 2-oxo-8alpha-hydroxyguaia-1(10),3,11(13)-trien-12,6alpha-olide, ketoplenolide B, alantolactone, 9beta-hydroxyeudesma-4,11(13)-dien-12-oic acid and 9beta-acetoxyeudesma-4,11(13)-dien-12-oic acid} . The structures of the three guaianolides were elucidated by a combination of spectroscopic methods (EIMS, HREIMS, COSY, HMQC, HMBC and NOESY) as 1beta,2beta-epoxy-3beta,4alpha,8beta,10alpha-tetrahydroxyguaia-11(13)-en-12,6alpha-olide (1), 1beta,2beta-epoxy-3beta,4alpha,9alpha,10alpha-tetrahydroxyguaia-11(13)-en-12,6alpha-olide (2) and 1beta,2beta-epoxy-10alpha-hydroperoxy-3beta,4alpha,8beta-trihydroxyguaia-11(13)-en-12,6alpha-olide (3), respectively . Antifungal bioassay of all isolates showed that guaianolides 1, 2, 3, and 1beta,2beta-epoxy-3beta,4alpha,10alpha-trihydroxyguaia-11(13)-en-12,6alpha-olide were inhibitory to the growth of Candida albicans with MICs being 20, 20, 20, and 40 microg/ml, respectively.

Trends Microbiol, 2001 Dec, 9(12), 591 - 6
Virulence in Candida species; Haynes K; Candida species other than Candida albicans now account for up to 50% of deep candidiasis cases, yet little attention has been paid to the virulence attributes of these fungi . Adherence to host tissues, response to environmental changes and the secretion of hydrolases are all thought to be important in Candida virulence . The identification of virulence attributes unique to a particular Candida species could provide powerful insights into the pathogenic process but will require the use of genome-wide approaches such as transcript profiling, signature-tagged mutagenesis and in vivo expression technology.

J Med Chem, 2001 Dec 6, 44(25), 4468 - 74
3-(Arylacetylamino)-N-methylbenzamides: a novel class of selective anti-Helicobacter pylori agents; Ando R et al.; After chemical modification preceded by the random screening of our chemical library, a novel class of selective anti-Helicobacter pylori agents was generated . Consequently, the 3-(arylacetylamino)-N-methylbenzamides, which were quite easy to prepare, showed potent inhibitory activity against Helicobacter pylori but exhibited no inhibitory activity against other sorts of bacteria and fungi, e.g., Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Bacteroides fragilis, and Candida albicans . These compounds showed potent anti-H . pylori activity under acidic conditions, whereas amoxicillin and clarithromycin decreased activity . The 3-(3-arylpropionylamino)-N-methylbenzamides, 3-(aryloxyacetylamino)-N-methylbenzamides, and (3-methylcarbamoylphenyl)carbamic acid 1-arylmethyl esters also exhibited potent anti-H . pylori activity . Finally, we selected 7n (BAS-118) as a candidate compound for further evaluation.

Scand J Infect Dis, 2001, 33(10), 749 - 51
Breakthrough candidaemias during empirical therapy with fluconazole in non-cancer and non-HIV adults caused by in vitro-susceptible Candida spp.: report of 33 cases; Kovacicova G et al.; The objective of this study was to assess risk factors and the outcome of breakthrough fungaemias (BFs) occurring during fluconazole (FLU) therapy in non-cancer and non-HIV individuals . Thirty-three fungaemias occurring during therapy with FLU among a total of 310 fungaemias observed within a 10-y national survey were analysed . The agar disk diffusion method was used for antifungal susceptibility testing and the Vitek system for species identification . Univariate and multivariate analysis was performed to determine risk factors for BF . All BFs were due to species known to be susceptible to FLU: Candida albicans (25/33), C . parapsilosis (6/33) and C . guillermondii (2/33) . The mean number of positive blood cultures per episode was 2.4 . The MIC of Candida spp . to FLU was 0.5-8 mg/ml (all strains were susceptible in vitro) . Neonatal age (< 4 weeks), very low birth weight, prior surgery, central venous catheter placement, artificial ventilation, total parenteral nutrition and C . parapsilosis were significantly related to BF in univariate analysis, but only central venous catheter placement was significantly related in multivariate analysis . However, the outcome of BFs and non-BFs was similar . All BFs occurred in non-HIV patients who were not previously treated with azoles, and were caused by in vitro FLU-susceptible species (C . albicans and C . tropicalis) . Thus factors other than in vitro susceptibility play a role in BFs.

Bratisl Lek Listy, 2001, 102(7), 314 - 7
Comparative study of disintegrated cells influence of Staphylococcus aureus, Escherichia coli and Candida albicans on human and mouse immune mechanisms; Bukovsky M et al.; The study presents comparison of immunomodulatory effects of Staphylococcus aureus, Escherichia coli and Candida albicans disintegrated cells on selected immune mechanisms of human and mouse leukocytes . We measured their phagocytic activity, phagocytic index and microbicidal activity against Staphylococcus aureus, Escherichia coli and Candida albicans cells as well as peroxidase and lysozyme activities of human and mouse leukocytes . Our results revealed predominantly inhibitory effect of disintegrated microorganisms on nonspecific immune functions of human leukocytes, but mainly stimulatory effect on mouse leukocytes monitored immune functions.

J Clin Microbiol, 2001 Dec, 39(12), 4309 - 15
Molecular characterization of new clinical isolates of Candida albicans and C . dubliniensis in Japan: analysis reveals a new genotype of C . albicans with group I intron; Tamura M et al.; The genetic diversity of recent clinical isolates of Candida albicans in Japan was studied on the basis of amplified DNA band lengths determined with a specific PCR primer reported to have been designed to span a transposable intron region in the 25S rRNA gene . Our analyses of 301 clinical isolates of C . albicans showed that they could be classified into five genotypes: genotype A (172 isolates), genotype B (66 isolates), genotype C (56 isolates), genotype D (C . dubliniensis; 5 isolates), and a new genotype (designated genotype E; 2 isolates) . The new genotype E was characterized to have a group I intron-like sequence, which is longer than hitherto reported ones and which has a nucleotide sequence length of 962 bp . Our analysis of the 962-bp sequence indicated that it is composed of an intron similar to that of C . dubliniensis of 621 bp with a 341-bp insertion . Analysis of the sequence of the internal transcribed spacer (ITS) region of the genotype E strain showed that its sequence is identical to those of strains of other genotypes, with only a few base substitution differences . Throughout the study, the possible horizontal transfer of the group I intron between C . dubliniensis and C . albicans was suggested . A high degree of correlation between the presence of a group I intron in C . albicans genotype E and susceptibility to the antifungal agent flucytosine was observed . The five isolates of C . dubliniensis examined in the present study showed genetic diversity when they were compared by randomly amplified polymorphic DNA fingerprinting pattern analysis, and this diversity was also confirmed by the analysis of ITS region sequences.

Pediatr Med Chir, 2001 May-Aug, 23(3-4), 197 - 9
{Favorable course of cerebral candidiasis in a low-birth newborn treated with liposomal amphotericin B}; Ferrari P et al.; A very low birth weight female infant developed a systemic infection by Candida albicans . Her clinical conditions were very poor; involvement of CNS was demonstrated my multifocal hyperechogenic images, spreading to the periventricular and subcortical areas of brain, confirmed by CT as circumscribed hyperdense punctiform granulations suggesting initial cerebritis . The introduction of liposomal Amphotericin-B in the therapy resulted in a marked improvement of clinical conditions together with the disapparance of pathological brain images . Longitudinal examinations carried out every three months during the first year, and every six months during the second, have shown adequate psychomotor development . The results of late cerebral nuclear magnetic resonance (NMR) imaging were within the norm . The patient is now seven-years-old and shows a well-balanced neuropsychological development . Systemic Candida infections of VLBW infants with CNS involvement have usually a very poor prognosis in terms of death or residual brain damage . The use of liposomal Amphotericin-B allowed us to achieve the complete cure of our patient, suggesting an high efficacy together with at a lower toxicity in respect of traditional treatments.

Curr Infect Dis Rep, 2001 Dec, 3(6), 546 - 549
Antimicrobial Resistance in Vulvovaginitis; Sobel JD; Although antimicrobial resistance has had an enormous impact on selection and utilization of antibiotics in virtually all aspects of clinical medicine, both inpatient and community based, little attention has been directed at antimicrobial resistance occurring in vaginal infections . Little evidence exists that frequent relapses of bacterial vaginosis or vulvovaginal candidiasis are due to antimicrobial resistance . Similarly, metronidazole-resistant trichomoniasis remains rare . Nevertheless, abuse of over-the-counter antimycotics, as well as widespread prescription of systemic oral azoles, could result in spread of azole-resistant Candida albicans, and even more likely could lead to an increase in non-albicans Candida species with intrinsic azole resistance . Problematic species include Candida glabrata and rarely Candida krusei.

Mol Microbiol, 2001 Nov, 42(3), 673 - 87
Ras links cellular morphogenesis to virulence by regulation of the MAP kinase and cAMP signalling pathways in the pathogenic fungus Candida albicans; Leberer E et al.; The pathogenic fungus Candida albicans is capable of responding to a wide variety of environmental cues with a morphological transition from a budding yeast to a polarized filamentous form . We demonstrate that the Ras homologue of C . albicans, CaRas1p, is required for this morphological transition and thereby contributes to the development of pathogenicity . However, CaRas1p is not required for cellular viability . Deletion of both alleles of the CaRAS1 gene caused in vitro defects in morphological transition that were reversed by either supplementing the growth media with cAMP or overexpressing components of the filament-inducing mitogen-activated protein (MAP) kinase cascade . The induction of filament-specific secreted aspartyl proteinases encoded by the SAP4-6 genes was blocked in the mutant cells . The defects in filament formation were also observed in situ after phagocytosis of C . albicans cells in a macrophage cell culture assay and, in vivo, after infection of kidneys in a mouse model for systemic candidiasis . In the macrophage assay, the mutant cells were less resistant to phagocytosis . Moreover, the defects in filament formation were associated with reduced virulence in the mouse model . These results indicate that, in response to environmental cues, CaRas1p is required for the regulation of both a MAP kinase signalling pathway and a cAMP signalling pathway . CaRas1p-dependent activation of these pathways contributes to the pathogenicity of C . albicans cells through the induction of polarized morphogenesis . These findings elucidate a new medically relevant role for Ras in cellular morphogenesis and virulence in an important human infectious disease.

FEMS Immunol Med Microbiol, 2001 Oct, 31(3), 211 - 7
Effects of interleukin-13 on antifungal activity of human monocytes against Candida albicans; Katsifa H et al.; We investigated the effects of human interleukin-13 (IL-13) on human monocytes' (MNC) activities against Candida albicans, an important human pathogen . Increased phagocytosis of blastoconidia was observed after incubation with 50 U ml(-1) of IL-13 for 4 h or 48 h in the presence or absence of serum . The latter effect was inhibited by anti-IL-13 monoclonal antibody or mannose . Incubation of MNC with 50 U ml(-1) of IL-13 for 2 h significantly enhanced superoxide anion production in response to phorbol myristate acetate . IL-13 did not, however, alter the damage caused by MNC to hyphae, whereas it suppressed killing of blastoconidia . IL-13 has variable effects on MNC activities and may play an important immunoregulatory role against C . albicans.

Int J Pharm, 2002 Jan 1, 231(1), 11 - 20
Microwave-assisted preparation of cyclic ketals from a cineole ketone as potential cosmetic ingredients: solvent-free synthesis, odour evaluation, in vitro cytotoxicity and antimicrobial assays; Genta MT et al.; Some cyclic ketals derived from (+)1,3,3-trimethyl-2-oxabicyclo{2.2.2}octan-6-one were obtained in excellent yields by microwave activation under solvent-free conditions, as a 'green chemistry' procedure . The results obtained using acidic alumina containing 7% p-toluenesulfonic acid, as mineral support, are reported and compared with those obtained by classical methods . The new compounds were tested for their olfactive character and for a potential cosmetic use . In vitro skin cytotoxicity tests were carried out on the most promising compounds, by using NCTC 2544 human keratinocytes as target cells . They all displayed slight cytotoxic effects which were one order of magnitude lower than those found with sodium dodecylsulphate positive control . Two compounds that resulted interesting as toothpaste aromas, were submitted to antimicrobial assays and showed their activity against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus hominis, Propionibacterium acnes and Candida albicans.

Proc Natl Acad Sci U S A, 2001 Dec 4, 98(25), 14637 - 42 Epub 2001 Nov 20.
The human salivary peptide histatin 5 exerts its antifungal activity through the formation of reactive oxygen species; Helmerhorst EJ et al.; Previous studies have shown that the human salivary antifungal peptide histatin 5 is taken up by Candida albicans cells and associates intracellularly with mitochondria . The purpose of the present study was to investigate the biological consequence of this specific subcellular targeting . Histatin 5 inhibited respiration of isolated C . albicans mitochondria as well as the respiration of intact blastoconidia in a dose and time-dependent manner . A nearly perfect correlation was observed between histatin-induced inhibition of respiration and cell killing with either logarithmic- or stationary-phase cells, but stationary-phase cells were less sensitive . Because nonrespiring yeast cells are insensitive to histatin 5, the potential mechanistic relationship between histatin 5 interference with the respiratory apparatus and cell killing was explored by using an oxygen radical sensitive probe (dihydroethidium) . Fluorimetric measurements showed that histatin 5 induced the formation of reactive oxygen species (ROS) in C . albicans cells as well as in isolated mitochondria and that ROS levels were highly correlated with cell death . In the presence of an oxygen scavenger (l-cysteine), cell killing and ROS formation were prevented . In addition, the membrane-permeant superoxide dismutase mimetic 2,2,6,6-tetramethylpiperidine-N-oxyl, abolished histatin-induced ROS formation in isolated mitochondria . In contrast to histatin 5, the conventional inhibitors of the respiratory chain, sodium cyanide or sodium azide, neither induced ROS nor killed yeast cells . These data provide strong evidence for a comprehensive mechanistic model of histatin-5-provoked yeast cell death in which oxygen radical formation is the ultimate and essential step.

J Bacteriol, 2001 Dec, 183(24), 7120 - 5
Novel posttranslational activation of the LYS2-encoded alpha-aminoadipate reductase for biosynthesis of lysine and site-directed mutational analysis of conserved amino acid residues in the activation domain of Candida albicans; Guo S et al.; The alpha-aminoadipate pathway for lysine biosynthesis is present only in fungi . The alpha-aminoadipate reductase (AAR) of this pathway catalyzes the conversion of alpha-aminoadipic acid to alpha-aminoadipic-delta-semialdehyde by a complex mechanism involving two gene products, Lys2p and Lys5p . The LYS2 and LYS5 genes encode, respectively, a 155-kDa inactive AAR and a 30-kDa phosphopantetheinyl transferase (PPTase) which transfers a phosphopantetheinyl group from coenzyme A (CoA) to Lys2p for the activation of Lys2p and AAR activity . In the present investigation, we have confirmed the posttranslational activation of the 150-kDa Lys2p of Candida albicans, a pathogenic yeast, in the presence of CoA and C . albicans lys2 mutant (CLD2) extract as a source of PPTase (Lys5p) . The recombinant Lys2p or CLD2 mutant extract exhibited no AAR activity with or without CoA . However, the recombinant 150-kDa Lys2p, when incubated with CLD2 extract and CoA, exhibited significant AAR activity compared to that of wild-type C . albicans CAI4 extract . The PPTase in the CLD2 extract was required only for the activation of Lys2p and not for AAR reaction . Site-directed mutational analysis of G882 and S884 of the Lys2p activation domain (LGGHSI) revealed no AAR activity, indicating that these two amino acids are essential for the activation . Replacement of other amino acid residues in the domain resulted in partial or full AAR activity . These results demonstrate the posttranslational activation and the requirement of specific amino acid residues in the activation domain of the AAR of C . albicans.

Arch Immunol Ther Exp (Warsz), 1998, 46(6), 381 - 6
TNF-alpha, IL-6 and IFN-gamma secreted by bronchoalveolar leukocytes isolated from patients with bronchial asthma, complicated by fungal airways infections; Cembrzynska-Nowak M et al.; It is widely known that fungal airways infections may deteriorate the course of bronchial asthma . The mechanism of the phenomenon is still unclear . The aim of our study was to assess the effect of fungal infections on the secretion of selected cytokines by bronchoalveolar leukocytes . Five patients (group FA) with bronchial asthma and Candida albicans or Aspergillus fumigatus airways infections (confirmed by bronchoscopy and culture) were included in the study . All of them were on the chronic treatment with corticosteroids (10-20 mg of prednisone per day) and underwent several courses of therapy with antibiotics . The control groups comprised 5 previously untreated asthmatics without bronchial colonization with fungi (group A) as well as 5 healthy volunteers (group H) . Leukocytes were isolated from bronchoalveolar lavage fluid (BALF) and cultured in the presence or absence of cytokine inducers such as phytohemagglutin L (PHA), lipo-polysaccharide (LPS) from E . coli . The activity of TNF-alpha, IL-6 and IFN-gamma were measured in the BAL cell culture supernatants by using specific bioassays . In comparison with healthy controls the spontaneous or induced secretion of cytokines were significantly augmented in patients from group A . In contrast the asthmatics who represented group FA demonstrated normal levels of spontaneous cytokine secretion . However, the tendency to increase LPS and PHA-induced production was observed in BAL leukocytes from the patients . The above results support the view that beneficial effect of corticosteroid treatment in bronchial asthma may act, at least in part, by inhibition of the high spontaneous secretion of proinflammatory cytokines . Nevertheless, fungal airways infections may lead to increase of LPS- or PHA-induced production of TNF-alpha, IL-6 or IFN-gamma (despite prednisone therapy) by prestimulation of the BAL cells with fungi.

J Antibiot (Tokyo), 2001 Sep, 54(9), 737 - 43
A novel assay for fungal ketol-isomerase activity; Nakata M et al.; 2-Deoxy-D-glucose-6-phosphate ketol-isomerase (EC 2.6.1.16) forms glucosamine-6-phosphate and glutamate from fructose-6-phosphate and glutamine and plays an important role in chitin synthesis in fungi . We have established a new assay for fungal ketol-isomerase activity that is amenable to high throughput screening to identify enzyme inhibitors . Aspergillus fumigatus crude lysate was incubated with substrates and after incubation, reactions were terminated . Glutamate dehydrogenase, nitro blue tetrazolium chloride, phenazine methosulfate and beta-NAD were added and the amount of glutamate formed by ketol-isomerase activity was determined by measuring OD585nm . A feedback inhibitor, UDP-N-acetylglucosamine, of fungal ketol-isomerase was successfully detected by this assay (IC50=0.48 mM) . In a pilot scale screening, an active extract from an extremophilic bacterium was found, and the extract showed antifungal activity against A . fumigatus, Candida albicans and C . glabrata.

Mycoses, 2001, 44(7-8), 281 - 6
Efficient treatment of murine systemic infection with Candida albicans using amphotericin B incorporated in nanosize range particles (emulsomes); Kretschmar M et al.; The effects of emulsome nanosize range lipid particles containing amphotericin B (EAmB) were compared with the reference formulation containing deoxycholate (Fungizone; Bristol-Myers Squibb, Munich, Germany) and with the commercial amphotericin lipid complex preparation (AmBisome; Nexstar, San Dimas, CA, USA) . The minimal inhibitory concentrations of Fungizone and EAmB were identical although killing of Candida albicans was delayed when EAmB was used . In a tissue culture model and in mice, the incorporation of AmB into emulsomes resulted in a considerable reduction of toxicity in comparison with Fungizone . For comparison of the in vivo effect of the preparations a mouse model of systemic infection with C . albicans was used . All preparations were able to reduce the fungal burden in the liver and kidneys in comparison with control animals treated with isotonic saline . AmBisome was more efficient in the reduction of the fungal burden of the liver than EAmB and Fungizone, even when applied in a similar dosage of 1 mg kg(-1) . In the kidneys, EAmB and Fungizone was slightly more effective than AmBisome . Therefore, in our models, the incorporation of AmB into nanosize particles was able to reduce toxicity without loss of efficiency . EAmB may be considered a candidate preparation for the treatment of infections with C . albicans in humans.

Mycoses, 2001, 44(7-8), 273 - 7
Oral colonization by Candida spp . among AIDS household contacts; Milan EP et al.; This study was designed to investigate the oral yeast colonization rate of household contacts of AIDS patients . Sixty-four AIDS household contacts were sequentially enrolled along with 103 HIV-negative blood bank donors (control group) . Samples were obtained by swabbing the oral mucosa . Yeast isolates were identified by classical methods and antifungal susceptibility testing was performed according to NCCLS microbroth assay . Candida spp . was recovered from the oral cavity of 33% of the AIDS household contacts, in contrast with 14% of the control group (P = 0.003 or P = 0.04 after adjusting for oral prosthesis use) . Candida albicans was the most frequently isolated species in both groups . All of the isolates were susceptible to fluconazole, itraconazole and ketoconazole . In conclusion, we were able to demonstrate a higher colonization rate in the AIDS household contacts group compared with the control group . No resistant isolates to antifungal drugs was observed . We suggest that the contact with AIDS patients may play a role as a risk factor for developing oral colonization by Candida spp.

Acta Pol Pharm, 2001 May-Jun, 58(3), 175 - 84
Synthesis of novel quinolines, pyranoquinolines, furoquinolines, thieno-quinoline and their effect on the ultrastructure of some pathogenic microorganisms; Ghorab MM et al.; New series of quinolines 2, 6, 7, 9, 14-17, 20, 21; pyrano{3,2-c}quinolines 3, 4, 5, 11; furo{3,2-c}quinolines 8, 18; oxazino{5,6-c}quinoline 19 and thieno{3,4-b}quinoline 22 were prepared and the impact of synthesized compounds on some pathogenic microorganisms isolated from clinical samples was studied . Compounds 4, 11 and 14 were found to be the most active against all tested microorganisms . The compound 14 which showed higher activity was selected to study its action on the ultrastructure of Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans . Death of numerous cells, destruction of the cell wall, cell contents poured into the medium and impairment of metabolism, were observed.

Ann R Australas Coll Dent Surg, 2000 Oct, 15, 286 - 91
Oral health status, oral microflora, and non-surgical periodontal treatment of renal transplant patients receiving cyclosporin A and FK506; Chu FC et al.; OBJECTIVES: To determine the oral health status, oral microflora and the effect of non-surgical periodontal treatment on gingival overgrowth of renal allograft recipients receiving either cyclosporin A (CsA) or FK506 (Tacrolimus) as an immunosuppressant . MATERIALS AND METHODS: A total of 47 patients receiving CsA (mean age 43.1 years) and 10 receiving FK506 (mean age 40.1 years) were included in the study . Stone casts were taken for measurement of gingival overgrowth . An oral rinse technique was used to investigate the prevalence of yeasts, and aerobic and facultatively anaerobic Gram-negative rods (AGNR) . RESULTS: The CsA and FK506 patients exhibited a Gingival Overgrowth Index (GOI) of 45.2%, and 25.1%, respectively (p < 0.05) . The CsA patients had a GOI of 15.2% after one year of non-surgical periodontal treatment . The difference between pre- and postoperative gingival overgrowth indices was significant (p < 0.0001) . Candida albicans and Klebsiella pneumoniae were the most notable yeast and AGRN found . CONCLUSIONS: Renal transplant patients, being immunocompromised, constitute a high-risk group for gingival overgrowth . However, the FK506 regime appeared to ameliorate this effect, compared with CsA . Non-surgical periodontal treatment was effective in reducing established gingival overgrowth in both CsA and FK506 patients (p < 0.05) . Adequate pre- and post-transplant oral health care is recommended, for these patients, irrespective of the drug regime.

Antimicrob Agents Chemother, 2001 Dec, 45(12), 3474 - 81
Quantitative PCR assay to measure Aspergillus fumigatus burden in a murine model of disseminated aspergillosis: demonstration of efficacy of caspofungin acetate; Bowman JC et al.; Caspofungin acetate (MK-0991) is an antifungal antibiotic that inhibits the synthesis of 1,3-beta-D-glucan, an essential component of the cell wall of several pathogenic fungi . Caspofungin acetate was recently approved for the treatment of invasive aspergillosis in patients who are refractory to or intolerant of other therapies . The activity of 1,3-beta-D-glucan synthesis inhibitors against Aspergillus fumigatus has been evaluated in animal models of pulmonary or disseminated disease by using prolongation of survival or reduction in tissue CFU as assay endpoints . Because these methods suffer from limited sensitivity or poor correlation with fungal growth, we have developed a quantitative PCR-based (qPCR) (TaqMan) assay to monitor disease progression and measure drug efficacy . A . fumigatus added to naive, uninfected kidneys as either ungerminated conidia or small germlings yielded a linear qPCR response over at least 4 orders of magnitude . In a murine model of disseminated aspergillosis, a burden of A . fumigatus was detected in each of five different organs at 4 days postinfection by the qPCR assay, and the mean fungal load in these organs was 1.2 to 3.5 log(10) units greater than mean values determined by CFU measurement . When used to monitor disease progression in infected mice, the qPCR assay detected an increase of nearly 4 log(10) conidial equivalents/g of kidney between days 1 and 4 following infection, with a peak fungal burden that coincided with the onset of significant mortality . Traditional CFU methodology detected only a marginal increase in fungal load in the same tissues . In contrast, when mice were infected with Candida albicans, which does not form true mycelia in tissues, quantitation of kidney burden by both qPCR and CFU assays was strongly correlated as the infection progressed . Finally, treatment of mice with induced disseminated aspergillosis with either caspofungin or amphotericin B reduced the A . fumigatus burden in infected kidneys to the limit of detection for the qPCR assay . Because of its much larger dynamic range, the qPCR assay is superior to traditional CFU determination for monitoring the progression of disseminated aspergillosis and evaluating the activity of antifungal antibiotics against A . fumigatus.

Antimicrob Agents Chemother, 2001 Dec, 45(12), 3433 - 6
Efficacy of ravuconazole in treatment of mucosal candidosis in SCID mice; Clemons KV et al.; A model of orogastric candidosis in SCID mice, which mimics disease seen in AIDS patients, was used to evaluate ravuconazole in comparison with fluconazole for treatment . Mice were infected orally with Candida albicans and received either no treatment or oral treatment once daily for 12 days with 1, 5, or 25 mg of ravuconazole per kg of body weight per day, 5 or 25 mg of fluconazole per kg per day, or diluent (10% dimethyl sulfoxide in 0.5% carboxymethyl cellulose) . The numbers of C . albicans CFU in the esophagus, stomach, small intestine, and cecum on day 25 in mice given no treatment and diluent were equivalent . Both doses of fluconazole significantly reduced numbers of CFU in all four tissues but were equivalent to each other . Ravuconazole showed dose-responsive improvement of clearance of CFU . Ravuconazole at 25 mg/kg was superior in reduction of numbers of CFU in all tissues to controls or 25 mg of fluconazole per kg and to other regimens in at least three tissues . Fluconazole at 25 mg/kg cured no infection in any tissue, whereas 25 mg of ravuconazole/kg cleared infection in all tissues from 50% of mice . Ravuconazole has good efficacy and the potential to cure mucosal candidosis in the absence of a functional immune response.

Antimicrob Agents Chemother, 2001 Dec, 45(12), 3416 - 21
MDR1-mediated drug resistance in Candida dubliniensis; Wirsching S et al.; Candida dubliniensis is a recently described opportunistic fungal pathogen that is closely related to Candida albicans . Candida dubliniensis readily develops resistance to the azole antifungal agent fluconazole, both in vitro and in infected patients, and this resistance is usually associated with upregulation of the CdMDR1 gene, encoding a multidrug efflux pump of the major facilitator superfamily . To determine the role of CdMDR1 in drug resistance in C . dubliniensis, we constructed an mdr1 null mutant from the fluconazole-resistant clinical isolate CM2, which overexpressed the CdMDR1 gene . Sequential deletion of both CdMDR1 alleles was performed by the MPA(R)-flipping method, which is based on the repeated use of a dominant mycophenolic acid resistance marker for selection of integrative transformants and its subsequent deletion from the genome by FLP-mediated, site-specific recombination . In comparison with its parental strain, the mdr1 mutant showed decreased resistance to fluconazole but not to the related drug ketoconazole . In addition, we found that CdMDR1 confers resistance to the structurally unrelated drugs 4-nitroquinoline-N-oxide, cerulenin, and brefeldin A, since the enhanced resistance to these compounds of the parent strain CM2 compared with the matched susceptible isolate CM1 was abolished in the mdr1 mutant . In contrast, CdMDR1 inactivation did not cause increased susceptibility to amorolfine, terbinafine, fluphenazine, and benomyl, although overexpression of CdMDR1 in a hypersusceptible Saccharomyces cerevisiae strain had previously been shown to confer resistance to these compounds . The effect of CdMDR1 inactivation was identical to that seen in two similarly constructed C . albicans mdr1 mutants . Therefore, despite species-specific differences in the amino acid sequences of the Mdr1 proteins, overexpression of CaMDR1 and CdMDR1 in clinical C . albicans and C . dubliniensis strains seems to confer the same drug resistance profile in both species.

Antimicrob Agents Chemother, 2001 Dec, 45(12), 3366 - 74
Functional expression of Candida albicans drug efflux pump Cdr1p in a Saccharomyces cerevisiae strain deficient in membrane transporters; Nakamura K et al.; Analysis of the transport functions of individual Candida albicans plasma membrane drug efflux pumps is hampered by the multitude of endogenous transporters . We have stably expressed C . albicans Cdr1p, the major pump implicated in multiple-drug-resistance phenotypes, from the genomic PDR5 locus in a Saccharomyces cerevisiae mutant (AD1-8u(-)) from which seven major transporters of the ATP-binding cassette (ABC) family have been deleted . High-level expression of Cdr1p, under the control of the S . cerevisiae PDR5 promoter and driven by S . cerevisiae Pdr1p transcriptional regulator mutation pdr1-3, was demonstrated by increased levels of mRNA transcription, increased levels of nucleoside triphosphatase activity, and immunodetection in plasma membrane fractions . S . cerevisiae AD1-8u(-) was hypersensitive to azole antifungals (the MICs at which 80% of cells were inhibited {MIC(80)s} were 0.625 microg/ml for fluconazole, <0.016 microg/ml for ketoconazole, and <0.016 microg/ml for itraconazole), whereas the strain (AD1002) that overexpressed C . albicans Cdr1p was resistant to azoles (MIC(80)s of fluconazole, ketoconazole, and itraconazole, 30, 0.5, and 4 microg/ml, respectively) . Drug resistance correlated with energy-dependent drug efflux . AD1002 demonstrated resistance to a variety of structurally unrelated chemicals which are potential drug pump substrates . The controlled overexpression of C . albicans Cdr1p in an S . cerevisiae background deficient in other pumps allows the functional analysis of pumping specificity and mechanisms of a major ABC transporter involved in drug efflux from an important human pathogen.

Antimicrob Agents Chemother, 2001 Dec, 45(12), 3304 - 9
Antifungal activities of two new azasordarins, GW471552 and GW471558, in experimental models of oral and vulvovaginal candidiasis in immunosuppressed rats; Martinez A et al.; Sordarins constitute a new class of antifungal agents with a novel mechanism of action involving the selective inhibition of fungal protein synthesis . A further evolution of this class of antifungals has led to a new family of sordarin derivatives called azasordarins . The therapeutic efficacies of two new azasordarins, GW471552 and GW471558, were studied in experimental models of oral and vulvovaginal candidiasis in immunosuppressed rats . In all cases rats were immunosuppressed with dexamethasone in the drinking water . Oral candidiasis was established by inoculating 0.1 ml of a yeast suspension containing 5 x 10(8) cells of Candida albicans 4711E with a cotton swab on three alternate days . Vulvovaginal candidiasis was established in ovariectomized and estrus-induced rats by intravaginal inoculation of 10(7) CFU of C . albicans 4711E in 0.1 ml of saline . GW471552 and GW471558 were administered at 1, 5, and 10 mg/kg of body weight via the subcutaneous route . In oral candidiasis, azasordarins were administered each 8 h for 7 consecutive days, while in vaginal candidiasis the compounds were given each 4 h for 3 consecutive days . Antifungal activity of azasordarins was assessed by colony counts and by histological examination 1 day after treatment . In the oral infection model, GW471552 and GW471558 administered at 5 mg/kg significantly reduced (P < 0.05) the number of CFU of C . albicans compared with untreated controls . In addition, GW471552 and GW471558 given at 10 mg/kg eradicated C . albicans from the oral cavities of 100% of infected animals . Against vulvovaginal infection, both compounds showed significant therapeutic efficacy . GW471552 was able to eradicate the vaginal fungal burden at a dose of 10 mg/kg, and it significantly reduced the number of CFU of C . albicans in vaginas of rats treated with a dose of 5 mg/kg (P < 0.05) . GW471558 showed greater efficacy, eradicating the fungal burden of 100% of infected rats at a dose of 5 mg/kg and significantly reducing (P < 0.05) the C . albicans vaginal counts even at a dose of 1 mg/kg . In both therapeutic efficacy studies, the histological findings confirmed the microbiological results . The experimental results presented show that the tested azasordarins are effective against oral and vulvovaginal candidiasis in immunosuppressed rats and could be promising antifungal agents for use in humans.

Braz J Biol, 2001 Aug, 61(3), 507 - 16 Epub 2002 Jan 28.
Differentiation and numerical analysis of oral yeasts based on SDS-Page profiles . Influence of the culture media on the whole-cell protein extracts; Hofling JF et al.; The application of gel electrophoresis and numerical analysis of yeast soluble proteins analysis to the investigation of 12 oral yeast strains belonging to five species is described . It involves one-dimensional electrophoresis of SDS-solubilized whole-cell proteins using different culture media for the cultivation of the cells, integration densitometries in the areas of the gels and percentages of the proteins extraction . These extracts were prepared from four isolates of Candida albicans, two of C . tropicalis, C . guilliermondii, C . parapsilosis and C . krusei . The extracts from whole-cells proteins using different culture media for the cultivation of the cells were fractionated by slab electrophoresis using a discontinuous buffer system . The corresponding patterns showed at least 36 polypeptides in the range of 14.4-200 kDa . Different isolates of each species were clearly different in each of the five species . The data obtained suggest that different nutritional compositions led to the expression of different proteins derived from alternatives metabolic pathways expressed by the electrophoretic profiles . The construction of a database of protein fingerprints and numerical analysis based on such data, may have some implications in the classification and identification of such species with epidemiological, ecological and taxonomic purposes . A well defined or synthetic culture media seems to be much properly.

Infect Immun, 2001 Dec, 69(12), 7898 - 903
Attenuation of virulence and changes in morphology in Candida albicans by disruption of the N-acetylglucosamine catabolic pathway; Singh P et al.; A Candida albicans mutant with mutations in the N-acetylglucosamine (GlcNAc) catabolic pathway gene cluster, including the GlcNAc-6-phosphate deacetylase (DAC1), glucosamine-6-phosphate deaminase (NAG1), and GlcNAc kinase (HXK1) genes, was not able to grow on amino sugars, exhibited highly attenuated virulence in a murine systemic candidiasis model, and was less adherent to human buccal epithelial cells in vitro . No germ tubes were formed by the mutant after induction with GlcNAc, but the mutant exhibited hyperfilamentation under stress-induced filamentation conditions . In addition, the GlcNAc catabolic pathway played a vital role in determining the colony phenotype . Our results imply that this pathway is very important because of its diverse links with pathways involved in virulence and morphogenesis of the organism.

Microbiology, 2001 Nov, 147(Pt 11), 3159 - 64
Antibodies to alpha5beta1 and alpha(v)beta3 integrins react with Candida albicans alcohol dehydrogenase; Klotz SA et al.; It has been hypothesized that Candida albicans possesses integrin-like receptors on its cell surface . This is because C . albicans binds numerous fluid-phase extracellular matrix (ECM) proteins on its cell surface and adheres to the same ECM proteins when immobilized . In addition, numerous antibodies to human integrins (receptors for ECM proteins) bind to the fungal cell surface and in so doing inhibit the binding of the respective proteins . To demonstrate the presence of such a cell surface integrin, a cDNA library of C . albicans yeast cells was screened with polyclonal antiserum to the human fibronectin receptor (alpha5beta1 integrin) . Clones isolated by this screening technique also reacted specifically to antiserum against the human vitronectin receptor (alpha(v)beta3 integrin) . DNA sequence analysis of the cloned insert predicted a 350 aa protein (37 kDa) . This predicted protein showed 75% homology at the nucleotide sequence level to alcohol dehydrogenase (ADH) of Saccharomyces cerevisiae . In vitro transcription/translation of the cloned inserts yielded a 37 kDa protein that was immunoprecipitated with antibodies to the alpha5beta1 and alpha(v)beta3 integrins and an antibody to a C . albicans fibronectin receptor . These antibodies and an mAb to the human vitronectin receptor demonstrated an antigen of -37 kDa present in the cell-wall preparations of C . albicans and in spent growth medium . All four antibodies reacted with authentic ADH . The possible significance of these results in relation to C . albicans adherence is discussed.

J Biol Chem, 2002 Feb 1, 277(5), 3440 - 6 Epub 2001 Nov 07.
The unique solution structure and immunochemistry of the Candida albicans beta -1,2-mannopyranan cell wall antigens; Nitz M et al.; Synthetic oligomers of the antigenic Candida albicans (1-->2)-beta-mannopyranans adopt a compact solution conformation that leads to numerous inter-residue nuclear Overhauser effects, including unprecedented nuclear Overhauser effects between n and n + 3 residues . In excellent agreement with experimentally determined distances, unrestrained molecular dynamics point to a single family of conformations that approximate a compact helical motif with a three-residue repeat for this unique homopolymer . When the synthetic di- to hexasaccharides were employed as inhibitors of monoclonal antibodies, which protect mice against a lethal dose of the yeast pathogen, a novel pattern of inhibitor activity was observed . Instead of the paradigm first reported by Kabat (Kabat, E . A . (1962) Fed . Proc . 21, 694-701; Kabat, E . A . (1966) J . Immunol . 97, 1-11), wherein homo-oligosaccharides exhibit increasing inhibitory activity with increasing size, here the maximum activity is reached for di- and trisaccharides and diminishes significantly for tetra-, penta-, and hexasaccharides . These immunochemical data correlate with the ordered conformation of the beta-1,2-linked mannopyranan and imply that a uniquely small antigenic determinant has potential as a component of synthetic conjugate vaccines against Candida albicans.

J Bacteriol, 2001 Dec, 183(23), 6917 - 23
Yeast PalA/AIP1/Alix homolog Rim20p associates with a PEST-like region and is required for its proteolytic cleavage; Xu W et al.; The Saccharomyces cerevisiae zinc finger protein Rim101p is activated by cleavage of its C-terminal region, which resembles PEST regions that confer susceptibility to proteolysis . Here we report that Rim20p, a member of the broadly conserved PalA/AIP1/Alix family, is required for Rim101p cleavage . Two-hybrid and coimmunoprecipitation assays indicate that Rim20p binds to Rim101p, and a two-hybrid assay shows that the Rim101p PEST-like region is sufficient for Rim20p binding . Rim101p-Rim20p interaction is conserved in Candida albicans, supporting the idea that interaction is functionally significant . Analysis of Rim20p mutant proteins indicates that some of its broadly conserved regions are required for processing of Rim101p and for stability of Rim20p itself but are not required for interaction with Rim101p . A recent genome-wide two-hybrid study (T . Ito, T . Chiba, R . Ozawa, M . Yoshida, M . Hattori, and Y . Sakaki, Proc . Natl . Acad . Sci . USA 98:4569-4574, 2000) indicates that Rim20p interacts with Snf7p and that Snf7p interacts with Rim13p, a cysteine protease required for Rim101p proteolysis . We suggest that Rim20p may serve as part of a scaffold that places Rim101p and Rim13p in close proximity.

J Bacteriol, 2001 Dec, 183(23), 6740 - 5
Characterization of Pneumocystis carinii PHR1, a pH-regulated gene important for cell wall Integrity; Kottom TJ et al.; Pneumocystis carinii remains an important opportunistic fungal pathogen causing life-threatening pneumonia in patients with AIDS and malignancy . Currently, little is known about how the organism adapts to environmental stresses and maintains its cellular integrity . We recently discovered an open reading frame approximately 600 bp downstream of the region coding GSC-1, a gene mediating beta-glucan cell wall synthesis in P . carinii . The predicted amino acid sequence of this new gene, termed P . carinii PHR1, exhibited 38% homology to Saccharomyces cerevisiae GAS1, a glycosylphosphatidylinositol-anchored protein essential to maintaining cell wall integrity, and 37% homology to Candida albicans PHR1/PHR2, pH-responsive genes encoding proteins recently implicated in cross-linking beta-1,3- and beta-1,6-glucans . In view of its homology to these related fungal genes, the pH-dependent expression of P . carinii PHR1 was examined . As in C . albicans, P . carinii PHR1 expression was repressed under acidic conditions but induced at neutral and more alkaline pH . PHR1-related proteins have been implicated in glucan cell wall stability under various environmental conditions . Although difficulties with P . carinii culture and transformation have traditionally limited assessment of gene function in the organism itself, we have successfully used heterologous expression of P . carinii genes in related fungi to address functional correlates of P . carinii-encoded proteins . Therefore, the potential role of P . carinii PHR1 in cell wall integrity was examined by assessing its ability to rescue an S . cerevisiae gas1 mutant with absent endogenous Phr1p-like activity . Interestingly, P . carinii PHR1 DNA successfully restored proliferation of S . cerevisiae gas1 mutants under lethal conditions of cell wall stress . These results indicate that P . carinii PHR1 encodes a protein responsive to environmental pH and capable of mediating fungal cell wall integrity.

Acta Paediatr, 2001 Oct, 90(10), 1196 - 8
Screening of rotavirus and adenovirus infections during prolonged hospitalization in a neonatal unit; Hallstrom M et al.; Rotavirus and adenovirus infections in 308 infants hospitalized for longer than 1 wk, and cases with necrotizing enterocolitis, were screened in a neonatal unit during a 15 mo period, covering two rotavirus epidemics in the community . Altogether, 1020 stool samples were collected weekly until hospital discharge, and in necrotizing enterocolitis cases at the onset of symptoms, and tested for rotavirus and adenovirus by means of enzyme-linked immunosorbent assay . The positive samples were further analysed by polymerase chain reaction . Enzyme-linked immunosorbent assay revealed five adenovirus-positive cases, which were tested negative by polymerase chain reaction . Out of 16 necrotizing enterocolitis cases, one was adenovirus- and another rotavirus positive when tested by polymerase chain reaction, the latter having a concomitant Candida albicans septicaemia . CONCLUSION: Routine rotavirus and adenovirus screening in hospitalized neonates seems to be unnecessary . Viral diagnostic examinations should be considered in patients with necrotizing enterocolitis.

J Enzyme Inhib, 2001, 16(3), 287 - 92
Inhibition of Saccharomyces cerevisiae phosphomannose isomerase by the NO-donor S-nitroso-acetyl-penicillamine; Salvati L et al.; Phosphomannose isomerase (PMI; EC . 5.3.1.8) is an essential metalloenzyme in the early steps of the protein glycosylation pathway in both prokaryotes and eukaryotes . The Cys150 residue (according to Candida albicans PMI numbering) is conserved in the active centre of mammalian and yeast PMI, but not in bacterial species where it is replaced by Asn . Here, the dose- and time-dependent inhibitory effect of the NO-donor S-nitroso-acetyl-penicillamine on the Saccharomyces cerevisiae PMI catalytic activity is reported . The analysis of the X-ray crystal structure of C . albicans PMI and of the molecular model of S . cerevisiae PMI provides a rationale for the low reactivity of Cys150 towards alkylating and nitrosylating agents.

Phytomedicine, 2001 Sep, 8(5), 389 - 94
Adesmia aegiceras: antimicrobial activity and chemical study; Agnese AM et al.; The antimicrobial activity of the ethanolic extract of Adesmia aegiceras was studied by the agar-well diffusion method . Antibacterial activity against Micrococcus luteus and eight pathogenic bacterial strains as well as antifungal activity against Candida albicans, was detected . Bacterial and fungal strains exhibited similar concentration-response curves (EC50 and Rmax values) and similar MIC . The MBC/MIC was about 8 . These data would indicate the potential usefulness of the A . aegiceras extract as a microbiostatic, antiseptic or disinfectant agent . Furthermore, chemical study of the bioactive alcoholic extract was performed, which revealed quercetin, isorhamnetin-3-rutinoside, isovitexin, pinitol and chlorogenic acid as its main components.

Med Oral, 2001 Nov-Dec, 6(5), 326 - 34
Oral-disease prevention in children with cancer: testing preventive protocol effectiveness; Rojas de Morales T et al.; Mucositis, gingivitis, herpetic stomatitis and candidiasis are a potential source of systemic infection in patients undergoing chemotherapy . Their severity and incidence may be reduced with procedures based on the prevention and elimination of sources causing oral infection and irritation . OBJECTIVE: The purpose of this investigation was to evaluate the effectiveness of an Oral Disease Preventive Protocol in children with cancer, subjected to chemotherapy and prior to application of dentobacterial infection control . MATERIAL AND METHODS: A controlled clinical test was run, with random assignations, on twelve 5-to-12-year-old patients diagnosed with Acute Lymphoblastic Leukemia (ALL) or Lymphoma, evaluated for twelve months, with a total of 154 evaluations . Five patients were boosted with oral physiotherapy, with non-alcoholic 0.05% fluoride mouthwashes, with topical application of myconazole oral gel; seven patients were given instructions on oral physiotherapy . RESULTS: There were no significant differences between the groups under evaluation (p>0.05) . Of the oral complications evaluated, gingivitis registered the highest percentage (60%), followed by mucositis (18%) and candida albicans infection (7%) . Most affected were the submandibular and cervical ganglions (59% and 41%, respectively) . CONCLUSIONS: Prior control of sources causing oral infection and irritation effectively prevents complications during non-surgical cancer therapy.

Respiration, 2001, 68(5), 465 - 70
Effects of amphotericin B gargles on oral colonization of Candida albicans in asthmatic patients on steroid inhalation therapy; Fukushima C et al.; BACKGROUND: Early use of inhaled steroids is recommended for bronchial asthma . The side effects are rare, but oral discomfort and candidiasis are clinically important complications . Most previous studies reported that the use of spacer and water gargling was necessary to prevent oral complications . However, in some patients, this may fail to prevent such complications . OBJECTIVE: To compare the effects of water gargling with those of amphotericin B, in the prevention of oral complications in asthmatics using inhaled steroids . METHODS: Pharyngeal swab samples were obtained aseptically from the posterior pharyngeal wall of 128 asthmatics who have been using inhaled steroids (beclomethasone dipropionate) for more than 1 year . The amount of Candida albicans in cultured swabs was evaluated based on the following criteria: oral symptoms, method of gargling, dose of inhaled steroids, type of spacer and serum cortisol level . RESULTS: The number of isolated C . albicans was significantly higher in asthmatics with oral symptoms than in those free of symptoms . It was also significantly higher in patients who gargled with water or 1,000 times dilution than in those who gargled with 100 or 50 times dilutions of amphotericin B . Moreover, it was significantly higher in patients with low levels of serum cortisol than in those with normal serum cortisol . CONCLUSION: We demonstrated that at least in a subgroup of asthmatics using steroid inhalers, gargling with water or even weak concentrations of amphotericin B does not prevent colonization of the throat with C . albicans . This group at high risk of developing oral candidiasis should gargle with amphotericin B at concentrations higher than 100 times dilution that can prevent clinically detectable oral candidiasis .

Mol Biol Cell, 2001 Nov, 12(11), 3631 - 43
Signaling through adenylyl cyclase is essential for hyphal growth and virulence in the pathogenic fungus Candida albicans; Rocha CR et al.; The human fungal pathogen Candida albicans switches from a budding yeast form to a polarized hyphal form in response to various external signals . This morphogenetic switching has been implicated in the development of pathogenicity . We have cloned the CaCDC35 gene encoding C . albicans adenylyl cyclase by functional complementation of the conditional growth defect of Saccharomyces cerevisiae cells with mutations in Ras1p and Ras2p . It has previously been shown that these Ras homologues regulate adenylyl cyclase in yeast . The C . albicans adenylyl cyclase is highly homologous to other fungal adenylyl cyclases but has less sequence similarity with the mammalian enzymes . C . albicans cells deleted for both alleles of CaCDC35 had no detectable cAMP levels, suggesting that this gene encodes the only adenylyl cyclase in C . albicans . The homozygous mutant cells were viable but grew more slowly than wild-type cells and were unable to switch from the yeast to the hyphal form under all environmental conditions that we analyzed in vitro . Moreover, this morphogenetic switch was completely blocked in mutant cells undergoing phagocytosis by macrophages . However, morphogenetic switching was restored by exogenous cAMP . On the basis of epistasis experiments, we propose that CaCdc35p acts downstream of the Ras homologue CaRas1p . These epistasis experiments also suggest that the putative transcription factor Efg1p and components of the hyphal-inducing MAP kinase pathway depend on the function of CaCdc35p in their ability to induce morphogenetic switching . Homozygous cacdc35 Delta cells were unable to establish vaginal infection in a mucosal membrane mouse model and were avirulent in a mouse model for systemic infections . These findings suggest that fungal adenylyl cyclases and other regulators of the cAMP signaling pathway may be useful targets for antifungal drugs.

Mol Biol Cell, 2001 Nov, 12(11), 3538 - 49
Candida albicans Int1p interacts with the septin ring in yeast and hyphal cells; Gale C et al.; The ability to switch between yeast and hyphal morphologies is an important virulence factor for the opportunistic pathogen Candida albicans . Although the kinetics of appearance of the filamentous ring that forms at the incipient septum differ in yeast and cells forming hyphae (germ tubes) (), the molecular mechanisms that regulate this difference are not known . Int1p, a C . albicans gene product with similarity in its C terminus to Saccharomyces cerevisiae Bud4p, has a role in hyphal morphogenesis . Here we report that in S . cerevisiae, Int1p expression results in the growth of highly polarized cells with delocalized chitin and defects in cytokinesis and bud-site selection patterns, phenotypes that are also seen in S . cerevisiae septin mutant strains . Expression of high levels of Int1p in S . cerevisiae generated elaborate spiral-like structures at the periphery of the polarized cells that contained septins and Int1p . In addition, Int1p coimmunoprecipitated with the Cdc11p and Cdc12p septins, and Cdc12p is required for the establishment and maintenance of these Int1p/septin spirals . Although Swe1p kinase contributes to INT1-induced filamentous growth in S . cerevisiae, it is not required for the formation of ectopic Int1p/septin structures . In C . albicans, Int1p was important for the axial budding pattern and colocalized with Cdc3p septin in a ring at the mother-bud neck of yeast and pseudohyphal cells . Under conditions that induce hyphae, both Cdc3p and Int1p localized to a ring distal to the junction of the mother cell and germ tube . Thus, placement of the Int1p/septin ring with respect to the mother-daughter cell junction distinguishes yeast/pseudohyphal growth from hyphal growth in C . albicans.

Mycopathologia, 2001, 152(1), 15 - 21
IgG and IgE immune response against the surface glycoprotein gp200 of Saccharomyces cerevisiae in patients with atopic dermatitis; Nenoff P et al.; The heat-stable and soluble glycoprotein gp200 (molecular weight 200 kDa) is part of the cell wall of S . cerevisiae . Recently, an association was shown between IgA and IgG against gp200 and inflammation in Crohn's disease . Gp200 is able to induce a proliferation of human lymphocytes in vitro, together with a natural killer cell associated cytotoxicity . Specific IgE against Saccharomyces cerevisiae (baker's or brewer's yeast) may be detected in approximately 73%, against Candida albicans in 68% of those patients suffering from severe atopic dermatitis . The aim of this study was to elucidate the possible role of an anti-gp200 immune response for the pathogenesis of atopic dermatitis by immunoblot analysis . Anti-gp200 IgE was found in 55% of healthy individuals, in 67% of individuals with atopic predisposition without eczema, in 63% of the patients with mild atopic dermatitis, and in 86% of patients with severe atopic dermatitis, respectively . On the contrary, anti-gp200 IgG could be shown in 55% of healthy individuals, in 89% of individuals with atopic predisposition but without eczema, in 100% of patients with mild atopic dermatitis, and in 79% with severe atopic dermatitis, respectively . No immunoreactivity was found when an extract of Arxula adeninivorans was used as antigen . These results underline the specificity of the immunoblot results with gp200 from Saccharomyces cerevisiae . It can be concluded that occurrence of specific IgE against Saccharomyces cerevisiae cannot be explained by a cross reactivity, e.g., against Candida albicans allergens . Further investigations with the recombinant gp200 will give information on the role of this glycoprotein both in atopic dermatitis and Morbus Crohn.

Biochem Biophys Res Commun, 2001 Nov 9, 288(4), 1001 - 5
Pseudin-2: an antimicrobial peptide with low hemolytic activity from the skin of the paradoxical frog; Olson L 3rd et al.; Four structurally related peptides (pseudins 1-4) with antimicrobial activity were isolated from an extract of the skin of the paradoxical frog Pseudis paradoxa (Pseudidae) . Pseudin-2 (GLNALKKVFQGIHEAIKLINNHVQ) was the most abundant peptide (22 nmol/g tissue) and also the most potent (minimum inhibitory concentrations, MIC = 2.5 microM against Escherichia coli, 80 microM against Staphylococcus aureus, and 130 microM against Candida albicans) . The concentration of pseudin-2 producing 50% hemolysis of human erythrocytes was >300 microM . Circular dichroism studies showed that the pseudins belong to the class of cationic, amphipathic alpha-helical antimicrobial peptides but their amino acid sequences are not similar to any previously characterized peptides from frog skin . The pseudins do, however, show sequence similarity with a region at the C-terminus of DEFT, a death effector domain-containing protein expressed in mammalian testicular germ cells that is involved in the regulation of apoptosis .

Clin Diagn Lab Immunol, 2001 Nov, 8(6), 1258 - 62
Role of alveolar macrophages in Candida-induced acute lung injury; Kubota Y et al.; Recent studies have shown that alveolar macrophages (AMs) not only act as phagocytes but also play a central role as potent secretory cells in various lung diseases, including pneumonia and acute respiratory distress syndrome . The behavior of AMs during disseminated candidiasis, however, is insufficiently elucidated . This study is the first to report disseminated candidiasis in AM-depleted mice and to analyze the effect of AMs on Candida-induced acute lung injury . While all AM-sufficient mice died by day 2 after infection with Candida albicans, no mortality was observed among AM-depleted mice . Unexpectedly, the CFU numbers of C . albicans isolated from the lungs of AM-depleted mice were significantly higher than those for C . albicans isolated from AM-sufficient mice . The lung wet-to-dry weight ratio was lower for AM-depleted mice than for AM-sufficient mice, although this difference was not significant . We found that bronchoalveolar lavage fluid (BALF) from AM-depleted mice in candidemia contained fewer neutrophils than BALF from AM-sufficient mice . In addition, myeloperoxidase activities in lung homogenates of AM-depleted mice were significantly lower than those in homogenates of AM-sufficient mice . A significant decrease in levels of murine macrophage inflammatory protein 2 (MIP-2), a potent chemoattractant for neutrophils, was noted in lung homogenates from AM-depleted mice compared with levels in homogenates from AM-sufficient mice . Immunohistochemical studies using anti-MIP-2 antibodies revealed that AMs were the cellular source of MIP-2 within the lung during candidemia . We observed that AM depletion decreased levels of AM-derived neutrophil chemoattractant, alleviated acute lung injury during candidemia, and prolonged the survival of mice in candidemia, even though clearance of C . albicans from the lungs was reduced.

Diagn Microbiol Infect Dis, 2001 Sep-Oct, 41(1-2), 23 - 7
Two-year surveillance on fluconazole susceptibility of Candida spp isolates in a general and university hospital in Rome; Testore GP et al.; Fluconazole susceptibility was tested in 385 clinical yeast isolates (285 Candida albicans, 38 C . glabrata, 31 C . tropicalis, 31 other Candida subsp.) using the agar disk diffusion test . Yeasts were collected from specimens obtained from outpatients (69) and inpatients (intensive care unit: 79 isolates, major burn unit: 31 isolates, hematology ward: 45 isolates, gynecology ward: 67 isolates, other wards: 94 isolates) . Three hundred and fifty-six (92%) yeast isolates showed to be susceptible, 18 (5%) were susceptible dose-dependent, and 10 (3%) were resistant to fluconazole . Of the resistant group, 3 isolates were C.albicans, while seven were Candida non-albicans (2 C . rugosa, 2 C . humicola, 1 C . tropicalis, 1 C . ciferrii, 1 C . glabrata) . The disk-diffusion method was easy to perform and there were no difficulties in the interpretation of inhibition zone diameters . Fluconazole maintained a good activity against Candida spp despite its extensive use for the prophylaxis and treatment of fungal infections.

J Immunol Methods, 2001 Nov 1, 257(1-2), 185 - 202
Recombinant human antibody single chain variable fragments reactive with Candida albicans surface antigens; Haidaris CG et al.; A combinatorial phage display library expressing human immunoglobulin heavy and light chain variable regions was used to identify phage clones capable of binding to the surface of Candida albicans blastoconidia . Single chain antibody variable fragments (scFv) derived from three clones detected C . albicans antigens by indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA), and Western blotting . The antigens detected were conserved among different strains of C . albicans and several other Candida species . Two scFv clones detected antigens specifically expressed by C . albicans blastoconidia; the third detected antigens in both blastoconidia and filamentous forms of C . albicans . The antigens containing the epitopes recognized by all three scFv could be extracted from blastoconidia by dithiothreitol, suggesting attachment to the cell wall via sulfhydryl bonds . Epitope detection by the scFv was sensitive to treatment of C . albicans blastoconidia with sodium periodate, but not proteinase K, indicating the cognate epitopes were composed of carbohydrate . Antigenic determinants for each of the three scFv were detected by immunohistochemical staining of skin sections from a model of cutaneous candidiasis, demonstrating expression in vivo . Through selection for the ability to bind intact organisms, the phage display system provides a means to rapidly identify monoclonal binding ligands to Candida surface antigens . Being entirely human, mature antibodies generated from the scFv have potential utility in the treatment of candidiasis.

Curr Microbiol, 2001 Dec, 43(6), 429 - 33
Factors affecting the morphogenetic switch in Yarrowia lipolytica; Perez-Campo FM et al.; Yarrowia lipolytica is a dimorphic yeast usually isolated from dairy products . Here we described methods for inducing in a homogeneous way a true yeast-hypha transition in liquid medium . As a first step, the cells must be synchronized in the G1 phase of the cell cycle by nitrogen starvation . Using either N-acetylglucosamine (GlcNAc) or serum as the only carbon sources, more than 90% of the cells form hypha after 4-6 h of incubation . Bovine albumin is also able to induce the yeast-hypha transition, although to a lesser extent . The addition of glucose to cultures growing with GlcNAc arrest the morphogenetic switch but not when added to cultures growing in the presence of serum . Serum also induces invasive growth in solid medium . Neither pH, nitrogen starvation, nor temperature play a relevant role in the morphogenetic switch . Our results suggest that, as occurs in Candida albicans, at least two morphogenetic signal pathways exist in Y . lipolytica.

J Clin Microbiol, 2001 Nov, 39(11), 4181 - 3
Trends in antifungal susceptibility among Swedish Candida species bloodstream isolates from 1994 to 1998: comparison of the E-test and the Sensititre YeastOne Colorimetric Antifungal Panel with the NCCLS M27-A reference method; Chryssanthou E; A comparative evaluation of the NCCLS macrodilution method, the E-test, and the Sensititre YeastOne Colorimetric Antifungal Panel for the susceptibility testing of fluconazole, itraconazole, amphotericin B, and flucytosine was conducted with 233 blood isolates of Candida species collected between 1994 and 1998 in Sweden . Antifungal susceptibility profiles of Candida albicans and non-C . albicans Candida species remained essentially unchanged within the 5-year study period . The overall agreement rates for the E-test and the NCCLS MICs and for the YeastOne and the NCCLS MICs were > or =86 and > or =87%, respectively, within +/-1 dilution for fluconazole, amphotericin B, and flucytosine, and > or =66 and > or =57%, respectively, for itraconazole . The E-test and the YeastOne panels are equivalent, and both are convenient methods for routine use.

J Clin Microbiol, 2001 Nov, 39(11), 4138 - 41
Differentiation of Candida albicans and Candida dubliniensis by fluorescent in situ hybridization with peptide nucleic acid probes; Oliveira K et al.; The recent discovery of Candida dubliniensis as a separate species that traditionally has been identified as Candida albicans has led to the development of a variety of biochemical and molecular methods for the differentiation of these two pathogenic yeasts . rRNA sequences are well-established phylogenetic markers, and probes targeting species-specific rRNA sequences have been used in diagnostic assays for the detection and identification of microorganisms . Peptide nucleic acid (PNA) is a DNA mimic with improved hybridization characteristics, and the neutral backbone of PNA probes offers significant advantages in whole-cell in situ hybridization assays . In this study, we developed PNA probes targeting the rRNAs of C . albicans and C . dubliniensis and applied them to a fluorescence in situ hybridization method (PNA FISH) for differentiation between C . albicans and C . dubliniensis . Liquid cultures were smeared onto microscope slides, heat fixed, and then hybridized for 30 min . Unhybridized PNA probe was removed by washing, and smears were examined by fluorescence microscopy . Evaluation of the PNA FISH method using smears of 79 C . dubliniensis and 70 C . albicans strains showed 100% sensitivity and 100% specificity for both PNA probes . We concluded that PNA FISH is a powerful tool for the differentiation of C . albicans and C . dubliniensis.

J Clin Microbiol, 2001 Nov, 39(11), 4076 - 81
Analysis of microsatellite markers of Candida albicans used for rapid typing; Botterel F et al.; To obtain a rapid genotyping method of Candida albicans, three polymorphic microsatellite markers were investigated by multiplex PCR . The three loci, called CDC3, EF3, and HIS3, were chosen because they are on different chromosomes so as to improve the chances of finding polymorphisms . One set of primers was designed for each locus, and one primer of each set was dye-labeled to read PCR signals by using an automatic sequencer . Amplifications were performed directly from the colonies harvested on the agar plate without a sophisticated DNA extraction step . At total of 27 reference strains and 73 clinical independent isolates were tested . The numbers of allelic associations were 10, 22, and 25 for the loci CDC3, EF3, and HIS3, respectively . The combined discriminatory power of the three microsatellites markers was 0.97 . The markers were stable after 25 subcultures, and the amplifications were specific for C . albicans . An initial study of 17 clinical isolate pairs, including blood culture and peripheral sites, showed a similar genotype for 15 of them, confirming that candidemia usually originates from the colonizing isolate . Therefore, microsatellite marker analysis with multiplex PCR and automated procedures has a high throughput and should be suitable for large epidemiologic studies of C . albicans.

J Clin Microbiol, 2001 Nov, 39(11), 4066 - 75
PCR fingerprinting of Candida albicans associated with chronic hyperplastic candidosis and other oral conditions; Bartie KL et al.; The purpose of this study was to genotype strains of Candida albicans to determine whether specific types were associated with chronic hyperplastic candidosis (CHC) . A total of 67 candidal isolates from CHC patients (n = 17) and from patients with other oral conditions (n = 21) were genotyped by PCR fingerprinting employing two interrepeat primer combinations (1245 and 1246 primers or 1251 primer) and a single minisatellite-specific M13 primer . The most suitable primer for fingerprint analysis was found to be primer 1251, yielding well-resolved banding patterns . For the 67 isolates tested, PCR fingerprinting delineated 25 (1245 and 1246 primers), 27 (1251 primer), and 25 (M13 primer) profiles . The majority of C . albicans isolates from multiple sites within the mouth produced identical profiles (six out of nine subjects examined) . For patients for whom a series of longitudinal isolates was available, strain persistence for up to 7 years was evident for five out of eight individuals, despite episodes of antifungal therapy . Computer-assisted comparison of the interrepeat PCR fingerprints identified seven distinct profiles that were shared among isolates from different individuals . However, no association was evident among isolates of C . albicans from specific clinical conditions . Eight isolates that were initially identified as C . albicans but having atypical PCR profiles were later confirmed as Candida dubliniensis . In conclusion, the genotypic data do not indicate clonal restriction of C . albicans with respect to CHC . Furthermore, these results have demonstrated that in the majority of individuals, colonizing populations of C . albicans are clonal in nature and exhibit strain persistence.

J Clin Microbiol, 2001 Nov, 39(11), 3830 - 7
Characterization of AFMP1: a novel target for serodiagnosis of aspergillosis; Yuen KY et al.; We cloned the AFMP1 gene, which encodes the first antigenic cell wall galactomannoprotein in Aspergillus fumigatus . AFMP1 codes for a protein, Afmp1p, of 284 amino acid residues, with a few sequence features that are present in Mp1p, the antigenic cell wall mannoprotein in Penicillium marneffei that we described previously, as well as several other cell wall proteins of Saccharomyces cerevisiae and Candida albicans . It contains a serine- and threonine-rich region for O glycosylation, a signal peptide, and a putative glycosylphosphatidyl inositol attachment signal sequence . Specific anti-Afmp1p antibody was generated with recombinant Afmp1p protein purified from Escherichia coli to allow further characterization of Afmp1p . Afmp1p has a high affinity for Galanthus nivalis agglutinin, a characteristic indicative of a mannoprotein . Furthermore, it was recognized by a rat monoclonal antibody against the galactofuran side chain of galactomannan, indicating that it is a galactomannoprotein . Ultrastructural analysis by immunogold staining indicated that Afmp1p is present in the cell walls of the hyphae and conidia of A . fumigatus . Finally, it was observed that patients with aspergilloma and invasive aspergillosis due to A . fumigatus develop a specific antibody response against Afmp1p . This suggested that the recombinant protein and its antibody may be useful for serodiagnosis in patients with aspergilloma or invasive aspergillosis, and the protein may represent a good cell surface target for host humoral immunity.

Eur J Clin Microbiol Infect Dis, 2001 Aug, 20(8), 544 - 53
Molecular monitoring of Candida albicans infections in liver transplant recipients; Tavanti A et al.; This report describes the use of the 27A probe for the molecular monitoring of Candida albicans infections in liver transplant recipients . Nosocomial candidiasis is the major fungal infection in liver transplant recipients, with Candida albicans being the species most frequently isolated . The molecular epidemiology of Candida albicans infections has been widely investigated, but scant attention has been focused on monitoring the identity of infecting strains in individual patients over the entire course of their hospitalization . In the study presented here, a total of 179 Candida albicans isolates were collected from 10 liver transplant recipients during multiple surveillance cultures performed before and after liver transplantation and from three healthcare workers at the Transplant Unit of Ospedale di Cisanello, Pisa (Italy) . Computer-aided analysis of the 27A-probed DNA fingerprints, used to compare the genetic relatedness of all the Candida albicans isolates, showed that most of the patients colonized with Candida albicans before transplantation harbored a unique Candida albicans genotype . This genotype persisted over the entire course of hospitalization and caused multiorgan failure in two patients, both of whom died from endogenously borne Candida albicans infections . Nosocomial acquisition of Candida albicans strains could be monitored in a timely manner in the other patients; for some of them, subsequent strain replacement was registered at different body sites during the post-transplant period . Neither cross-infection between patients nor transmission from healthcare workers to patients occurred in this hospital setting . These results indicate that the molecular monitoring of Candida albicans strains isolated from liver transplant recipients during their hospitalization may provide timely information about the identity of individual Candida albicans strains causing infections.

Proteomics, 2001 Apr, 1(4), 550 - 9
Analysis of the serologic response to systemic Candida albicans infection in a murine model; Pitarch A et al.; Two different strains of mice with different susceptibilities to systemic candidiasis (BALB/c and CBA/H) were infected with Candida albicans SC5314 . Immune sera were obtained on different days post-infection and assayed against two-dimensional polyacrylamide gel electrophoresis separation of cytoplasmic extracts obtained from protoplasts . More than 31 immunoreactive proteins were detected . Some of them were identified and found to correspond to (i) glycolytic enzymes, such as fructose biphosphate aldolase, triose phosphate isomerase (TPIS), glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase (PGK), enolase (ENO1) and pyruvate kinase, (ii) other metabolic enzymes, such as methionine synthase (METE), inosine-5'-monophosphate dehydrogenase (IMH3), alcohol dehydrogenase and aconitate hydratase and (iii) heat shock proteins: HS71 (or Ssa1p) and HS75 (or Ssb1p), both from the HSP70 family . This work reports for the first time antigenic properties for IMH3 and TPIS . Different profiles of antibody expression, depending on the mouse strain and the course of infection, were observed . ENO1 was the most immunogenic protein in infected BALB/c mice (the most resistant strain) . On the other hand, sera from CBA/H mice (a more susceptible strain) showed a strong increase in reactivity along the infection against METE, HS75 and PGK . Many of these immunoreactive proteins have also been detected using sera from human patients with systemic candidiasis, thus indicating the usefulness of the murine model for studying the antibody response in systemic candidiasis . In this work we demonstrate that the combination of two-dimensional electrophoresis with immunoblotting using murine immune sera can be an important tool for the identification of C . albicans antigens and for monitoring the evolution of the disease.

Boll Chim Farm, 2001 Sep-Oct, 140(5), 297 - 301
Synthesis and antimicrobial evaluation of chalcone and syndrome derivatives of 4(3H)-quinazolinone; Bekhit AA et al.; The increasing clinical importance of drug-resistant bacterial pathogens has encouraged additional microbiological and antibacterial research . New chalcone and sydnone derivatives of 4(3H)-quinazolinone were synthesized and evaluated for their antibacterial and antifungal activity . The microorganisms used were Escherichia coli ATCC 25922 as Gram-negative bacteria, Staphylococcus aureus ATCC 19433 as Gram-Positive bacteria and Candida albicans as yeast like fungi . The most potent compound was the nitroso derivative 6b, which exhibits interesting antibacterial and antifungal activities.

Science, 2001 Oct 26, 294(5543), 870 - 5
The plasticity of dendritic cell responses to pathogens and their components; Huang Q et al.; Dendritic cells are involved in the initiation of both innate and adaptive immunity . To systematically explore how dendritic cells modulate the immune system in response to different pathogens, we used oligonucleotide microarrays to measure gene expression profiles of dendritic cells in response to Escherichia coli, Candida albicans, and influenza virus as well as to their molecular components . Both a shared core response and pathogen-specific programs of gene expression were observed upon exposure to each of these pathogens . These results reveal that dendritic cells sense diverse pathogens and elicit tailored pathogen-specific immune responses.

J Antimicrob Chemother, 2001 Nov, 48(5), 713 - 5
Effect of the growth medium on the in vitro antifungal activity of micafungin (FK-463) against clinical isolates of Candida dubliniensis; Muller FM et al.; Micafungin (FK-463), a member of the new candin family of antifungal agents, was highly active against clinical isolates of Candida albicans and Candida dubliniensis . The in vitro activity of micafungin suggested that it was more potent than fluconazole, flucytosine, amphotericin B or voriconazole against C . albicans, and comparable or moderately less effective against C . dubliniensis isolates when high-resolution medium (HR) was used . Lower MICs of micafungin were recorded when RPMI 2% or AM3 2% media were used, indicating an influence of the growth medium on the MIC.

J Nat Prod, 2001 Oct, 64(10), 1282 - 5
Phenolic compounds from Miconia myriantha inhibiting Candida aspartic proteases; Li XC et al.; Assay-guided fractionation of the ethanol extract of the twigs and leaves of Miconia myriantha yielded two new compounds, mattucinol-7-O-{4' ',6' '-O-(S)-hexahydroxydiphenoyl}-beta-D-glucopyranoside (1) and mattucinol-7-O-{4' ',6' '-di-O-galloyl}-beta-D-glucopyranoside (2), along with mattucinol-7-O-beta-D-glucopyranoside (3), ellagic acid (4), 3,3'-di-O-methyl ellagic acid-4-O-beta-D-xylopyranoside, and gallic acid . Complete (1)H and (13)C NMR assignments of compound 1, which possesses a hexahydroxydiphenoyl unit, were achieved using the HMBC technique optimized for small couplings to enhance the four-bond and two-bond H/C correlations . Compounds 1 and 4 showed inhibitory effects against Candida albicans secreted aspartic proteases, with IC(50) of 8.4 and 10.5 microM, respectively.

Clin Oral Investig, 2001 Sep, 5(3), 172 - 6
Effect of antimicrobial mouthrinses on the in vitro adhesion of Candida albicans to human buccal epithelial cells; Pizzo G et al.; Adhesion to epithelial cells is a critical step in successful oral colonization and infection by Candida albicans . Therefore, three mouthrinse products, containing chlorhexidine 0.2% (CHX), cetylpyridinium chloride 0.05% (CPC) or triclosan 0.045% (TRN), were compared for their effects on the in vitro adhesion of C . albicans to human buccal epithelial cells (BEC) . Candidal adhesion appeared to be significantly reduced by oral rinsing with the CHX-containing mouthrinse (P<0.0001) . In vivo exposure of BEC to the CPC mouthrinse also inhibited adhesion of C . albicans (P<0.0001) . Both CHX and CPC products suppressed adhesion to the same extent (P>0.01) . On the other hand, the TRN mouthrinse did not significantly affect epithelial adhesion of C . albicans (P>0.01) . These findings suggest that mouthrinses containing CHX or CPC could be of value in the control of candidal colonization and infection . Clinical trials are warranted on the effectiveness of these products in reducing oral Candida carriage.

Biol Neonate, 2001, 80(4), 251 - 6
Effects of macrophage colony-stimulating factor on antifungal activity of neonatal monocytes against Candida albicans; Gioulekas E et al.; The effects of recombinant macrophage-colony stimulating factor (M-CSF) on antifungal activities of monocytes (MNC) from healthy neonates and adults against Candida albicans were compared . Pretreatment of adult and neonatal MNC with 15 ng/ml of M-CSF for 4 days significantly increased superoxide anion (O(-2)) production in response to phorbol myristate acetate . While M-CSF-treated MNC from adults produced significantly higher O(-2) in response to Candida blastoconidia, M-CSF-treated neonatal MNC did not show a similar response . Further, M-CSF significantly enhanced phagocytosis of C . albicans by adult MNC but not by neonatal MNC . While M-CSF enhances antifungal activities of adult MNC against C . albicans, it does not appear to affect anticandidal function of neonatal MNC .

Arthroscopy, 2001 Oct, 17(8), 878 - 83
Infection following knee arthroscopy; Wind WM et al.; Arthroscopy of the knee is not a risk-free procedure . Although rare, numerous complications have been reported in the literature . Fortunately, infection is a rare complication following arthroscopy, which, when treated, usually results in a benign outcome . We present the first reported case of Candida albicans infection following routine arthroscopy of the knee, which eventually resulted in a knee fusion . A review of infections that can occur after knee arthroscopy and their treatment is also presented . This and other potential complications should be considered when performing knee arthroscopy.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3209 - 12
In vitro killing of Candida albicans by fatty acids and monoglycerides; Bergsson G et al.; The susceptibility of Candida albicans to several fatty acids and their 1-monoglycerides was tested with a short inactivation time, and ultrathin sections were studied by transmission electron microscopy (TEM) after treatment with capric acid . The results show that capric acid, a 10-carbon saturated fatty acid, causes the fastest and most effective killing of all three strains of C . albicans tested, leaving the cytoplasm disorganized and shrunken because of a disrupted or disintegrated plasma membrane . Lauric acid, a 12-carbon saturated fatty acid, was the most active at lower concentrations and after a longer incubation time.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3195 - 7
New model of oropharyngeal candidiasis in mice; Kamai Y et al.; We established a straightforward murine model of oropharyngeal candidiasis . Mice were immunosuppressed with cortisone acetate, anesthetized, and then inoculated by placing cotton wool balls saturated with Candida albicans sublingually for 2 h . A prolonged, reproducible infection was induced . This model may be useful for antifungal screening or pathogenesis studies.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3037 - 45
In vitro and in vivo effects of 14alpha-demethylase (ERG11) depletion in Candida glabrata; Nakayama H et al.; Sterol 14alpha-demethylase (ERG11) is the target enzyme of azole antifungals that are widely used for the treatment of fungal infections . Candida glabrata is known to be less susceptible to fluconazole than most Candida albicans strains, and the incidence of C . glabrata infection has been increasing mostly in conjunction with the use of azole antifungals . Recently, it has been reported that C . glabrata can rescue the defect of ergosterol biosynthesis by incorporating cholesterol from serum . To explore the effect of inactivating Erg11p in C . glabrata, we generated mutant strains in which the ERG11 gene was placed under the control of tetracycline-regulatable promoters . In these mutants, expression of the ERG11 gene can be repressed by doxycycline (DOX) . All mutants showed a growth defect in the presence of DOX . The numbers of CFU of the mutants were lowered by only 1/10 with DOX treatment . In these mutants, accumulation of 4,14-dimethylzymosterol, which differs from an accumulated abnormal sterol detected in C . albicans and Saccharomyces cerevisiae treated with fluconazole, was observed by DOX treatment . Although such phenotypes were also observed in serum-containing media by DOX treatment, they were alleviated . Furthermore, the mutant could grow in DOX-treated mice without a severe reduction in the number of cells . Thus, depleting the expression of the ERG11 gene lowered the number of CFU by only 1/10 due to the accumulation of 4,14-demethylzymosterol in vitro, and it did not result in the defective growth of fungal cells in mice . These results suggested that Erg11p is not an ideal target molecule of antifungals for C . glabrata.

J Infect Dis, 2001 Nov 1, 184(9), 1170 - 5 Epub 2001 Oct 12.
Adherence and invasion studies of Candida albicans strains, using in vitro models of esophageal candidiasis; Bernhardt J et al.; The adherence of clinical and commensal isolates and reference collection strains of Candida albicans to a human esophageal cell monolayer (HET1-A) and reconstituted human esophageal tissue was compared . Isolates from patients with a severe form of esophageal candidiasis or candidemia adhered to HET1-A cells to a significantly greater extent than did isolates from patients with mild esophageal candidiasis or commensal and reference collection strains . In addition, C . albicans strain SSK21, which lacks the ssk1 response regulator gene of a 2-component signal transduction pathway, adhered less readily to the HET1-A cells than did parental cells or a gene-reconstituted strain . In a reconstituted esophageal tissue model, all clinical strains but not commensal or reference collection strains penetrated the epithelium, albeit at different rates . Hyphal formation following yeast cell adherence to the esophageal tissue was a requirement for invasion . Scanning electron microscopy was also used to confirm the colonization of the esophageal tissues by various strains . These studies indicate that both the HET1-A and the reconstituted esophageal tissue models can be used as in vitro targets to evaluate the adherence phenotype and invasiveness of C . albicans strains.

Infect Immun, 2001 Nov, 69(11), 7162 - 4
Resistance of T-cell receptor delta-chain-deficient mice to experimental Candida albicans vaginitis; Wormley FL Jr et al.; Conditions consistent with tolerance or immunoregulation have been observed in experimental Candida albicans vaginal infections . The present study investigated the role of gamma/delta T cells in experimental vaginal candidiasis . Results showed that T-cell receptor delta-chain-knockout mice had significantly less vaginal fungal burden when compared to wild-type mice, suggesting an immunoregulatory role for gamma/delta T cells in Candida vaginitis.

Infect Immun, 2001 Nov, 69(11), 7091 - 9
Potential role for a carbohydrate moiety in anti-Candida activity of human oral epithelial cells; Steele C et al.; Candida albicans is both a commensal and a pathogen at the oral mucosa . Although an intricate network of host defense mechanisms are expected for protection against oropharyngeal candidiasis, anti-Candida host defense mechanisms at the oral mucosa are poorly understood . Our laboratory recently showed that primary epithelial cells from human oral mucosa, as well as an oral epithelial cell line, inhibit the growth of blastoconidia and/or hyphal phases of several Candida species in vitro with a requirement for cell contact and with no demonstrable role for soluble factors . In the present study, we show that oral epithelial cell-mediated anti-Candida activity is resistant to gamma-irradiation and is not mediated by phagocytosis, nitric oxide, hydrogen peroxide, and superoxide oxidative inhibitory pathways or by nonoxidative components such as soluble defensin and calprotectin peptides . In contrast, epithelial cell-mediated anti-Candida activity was sensitive to heat, paraformaldehyde fixation, and detergents, but these treatments were accompanied by a significant loss in epithelial cell viability . Treatments that removed existing membrane protein or lipid moieties in the presence or absence of protein synthesis inhibitors had no effect on epithelial cell inhibitory activity . In contrast, the epithelial cell-mediated anti-Candida activity was abrogated after treatment of the epithelial cells with periodic acid, suggesting a role for carbohydrates . Adherence of C . albicans to oral epithelial cells was unaffected, indicating that the carbohydrate moiety is exclusively associated with the growth inhibition activity . Subsequent studies that evaluated specific membrane carbohydrate moieties, however, showed no role for sulfated polysaccharides, sialic acid residues, or glucose- and mannose-containing carbohydrates . These results suggest that oral epithelial cell-mediated anti-Candida activity occurs exclusively with viable epithelial cells through contact with C . albicans by an as-yet-undefined carbohydrate moiety.

Infect Immun, 2001 Nov, 69(11), 6813 - 22
Candida albicans is phagocytosed, killed, and processed for antigen presentation by human dendritic cells; Newman SL et al.; Candida albicans is a component of the normal flora of the alimentary tract and also is found on the mucocutaneous membranes of the healthy host . Candida is the leading cause of invasive fungal disease in premature infants, diabetics, and surgical patients, and of oropharyngeal disease in AIDS patients . As the induction of cell-mediated immunity to Candida is of critical importance in host defense, we sought to determine whether human dendritic cells (DC) could phagocytose and degrade Candida and subsequently present Candida antigens to T cells . Immature DC obtained by culture of human monocytes in the presence of granulocyte-macrophage colony-stimulating factor and interleukin-4 phagocytosed unopsonized Candida in a time-dependent manner, and phagocytosis was not enhanced by opsonization of Candida in serum . Like macrophages (Mphi), DC recognized Candida by the mannose-fucose receptor . Upon ingestion, DC killed Candida as efficiently as human Mphi, and fungicidal activity was not enhanced by the presence of fresh serum . Although phagocytosis of Candida by DC stimulated the production of superoxide anion, inhibitors of the respiratory burst (or NO production) did not inhibit killing of Candida, even when phagocytosis was blocked by preincubation of DC with cytochalasin D . Further, although apparently only modest phagolysosomal fusion occurred upon DC phagocytosis of Candida, killing of Candida under anaerobic conditions was almost equivalent to killing under aerobic conditions . Finally, DC stimulated Candida-specific lymphocyte proliferation in a concentration-dependent manner after phagocytosis of both viable and heat-killed Candida cells . These data suggest that, in vivo, such interactions between DC and C . albicans may facilitate the induction of cell-mediated immunity.

J Biol Chem, 2001 Dec 28, 276(52), 48988 - 96 Epub 2001 Oct 10.
DNA array studies demonstrate convergent regulation of virulence factors by Cph1, Cph2, and Efg1 in Candida albicans; Lane S et al.; Candida albicans, normally a human commensal, can cause fatal systemic infections under certain circumstances . Its unique ability to switch from yeast to hyphal growth in response to various environmental signals is inherent to its pathogenicity . Filamentation is regulated by multiple pathways including a Cph1-mediated mitogen-activated protein kinase pathway, an Efg1-mediated cAMP/PKA pathway, and a Cph2 pathway . To gain a general picture of how these various signaling pathways regulate differential gene expression during filamentation, we have constructed a partial C . albicans DNA array of 7,000 genes and used it to study the gene expression profiles using various mutants and growth conditions . By combining this novel technology with a new liquid medium in which cph1/cph1 is defective in filamentation, previously identified differentially expressed genes (ECE1, HWP1, HYR1, RBT1, SAPs5-6, and RBT4) are found to be regulated by all three pathways . In addition, two novel genes, DDR48 and YPL184, have been found to be differentially regulated during hyphal development and by all three pathways . This suggests that distinct filamentation signaling pathways converge to regulate a common set of differentially expressed genes . As one of the mechanisms for the observed convergence, we find that the transcription of a key regulator, TEC1, is regulated by Efg1 and Cph2 . Importantly, most of the genes regulated by multiple filamentation pathways encode known virulence factors . Perhaps, C . albicans utilizes converging pathways to regulate its vital virulence factors to ensure its survival and pathogenicity in various host environments.

Chin Med J (Engl), 1999 May, 112(5), 466 - 71
Resistant mechanisms of Candida albicans to fluconazole; Wang W et al.; OBJECTIVE: To detect the resistant mechanisms of Candida albicans to fluconazole (FCZ) at molecular biology level, since the resistance mechanisms of azole antifungal agents have been the focus of attention these years . METHODS: Thirty-two FCZ-resistant C . albicans were selected as our test strains (MICs > or = 64 micrograms/ml) . With 14-alpha-demethylase gene (ERG16 gene, target enzyme encoding gene of azoles) as our object, we chose six sets of primers from the ERG16 gene to amplify the interested fragments, and conducted Southern blot hybridization, restriction fragment length polymorphism (RFLP), and single strand conformation polymorphism (SSCP) analysis for the fragments which were amplified by the six sets of primers, and pre-resistant sensitive strains were used as controls . Three representative fragments, A66, D66 and E78, were selected to be cloned and sequenced . RESULTS: The polymerase chain reaction (PCR) amplification showed that several tested strains were negative for some primers . However, our Southern blot analysis reminded that their resistance did not result from the lack of target enzyme coding gene . SSCP analysis showed that differences were noted between the resistant and sensitive strains and inter-resistant strains . Statistical analysis showed that the most variable sequence lied in the amplifier of the sixth pair of primer, and all the tested 32 strains showed positive results . In the 11 mutation points we found, five resulted in amino acid alterations . It is likely that one or more mutational alterations (alone or in combination) might lead to the expression of an enzyme highly resistant to the inhibitory action of FCZ which in turn is responsible for the FCZ resistant trait in these strains . CONCLUSION: One or more mutational alterations might lead to the azole resistant trait in this strains.

Aust N Z J Obstet Gynaecol, 2001 Aug, 41(3), 326 - 8
Oestrogen, glycogen and vaginal candidiasis; Dennerstein GJ et al.; Our aim was to relate vaginal candidiasis to vaginal oestrogenisation . First, the incidence was determined (subjected to chi-square analysis) of vaginal Candida albicans infection in 339 consecutive dermogynaecology clinic patients aged 55 years and over, of whom 142 were using and 197 were not using oestrogen . Second, the ability of Candida species to utilise glycogen as a sole nutrient source was studied by performing assimilation tests using yeast nitrogen broth as a basal medium . infection on initial presentation compared with 4% in the cohort not using oestrogen (p < 0.001) . All 34 isolates of C . albicans assimilated glycogen . Twenty-six non-albicans species of Candida tested did not assimilate glycogen . In this study of postmenopausal women, there was a highly significant relationship between the usage of oestrogen and the occurrence of C . albicans infection . The production of glycogen by oestrogen stimulated epithelial maturation provides an attractive substrate for C . albicans.

Ginecol Obstet Mex, 2001 Jul, 69, 272 - 6
{Frequency of Gardnerella vaginalis vaginosis and its association with other pathogens causing genital infections in the female}; Mendoza-Gonzalez A et al.; In order to determine the presence of unspecific vaginosis and their causes, 700 vaginal smears were obtained from patients assisting to the Clinical Laboratory of the Familiar Medicine Unit No . 28 "Gabriel Mancera" of the Instituto Mexicano del Seguro Social, during 6 months . The patients age was from 18 to 55 years old . To establish the infectious etiology of these pathologies the vaginal smears were observed freshly and stained by Gram's method . Specific test for differentiate Gardnerella vaginalis and Candida albicans were also performed . From 700 vaginal smears, 160 were positive to Candida albicans (22.86%); 150 to Gardnerella vaginalis (21.43%); and 14 to Trichomonas vaginalis (2%) . The most frequent association were Candida albicans with Gardnerella vaginalis in 14 women, who 12 had a reduced number of pregnancy, and 7 had only one pregnancy (58.3%) . The age groups most affected were between 18 and 35 years old, corresponding to the reproductive stage of the woman . Gardnerella vaginalis predominated in the 30-35 years old group.

Peptides, 2001 Oct, 22(10), 1669 - 74
The effect of charge increase on the specificity and activity of a short antimicrobial peptide; Hong SY et al.; By using short linear antimicrobial peptides as a model system, the effect of peptide charge on the specificity between Candida albicans (fungi) and Gram-positive bacteria was investigated . In a present study, we added and/or deleted lysine residue(s) at the C-terminal and/or N-terminal end(s) of an antimicrobial peptide (KKVVFKVKFK-NH(2)) and synthesized the peptides that had similar alpha helical structures in a lipid membrane mimic condition . The increase of peptide charge improved antifungal activity without the change of antibacterial activity . Structure-activity relationship study about the peptides revealed that the net positive charge must play an important role in the specificity between C . albicans and Gram-positive bacteria and the increase of the net positive charge without the moderate change of secondary structure could improve activity for C . albicans rather than Gram-positive bacteria.

J Ethnopharmacol, 2001 Nov, 78(1), 1 - 5
Enhanced antifungal activity of ketoconazole by Euphorbia characias latex against Candida albicans; Giordani R et al.; The in vitro suseptibility of Candida albicans to ketoconazole and Euphorbia characias latex alone or in combination was tested using the macrobroth dilution method . The MIC 80% of crude latex and ketoconazole are respectively 159 microg protein/ml and 0.3901 microg/ml . This method permits us to determine an affinity constant K(aff) for crude latex (0.015 microg(-1) protein ml) and ketoconazole (23.828 microg(-1) ml) . The utilization of a mixture of latex at several concentrations (7.8-15.62-31.25-62.5 and 125 microg protein/ml) and ketoconazole indicates a synergistic effect between latex and ketoconazole . For latex concentrations of 31.25 and 62.5 microg protein/ml the MIC 80% of ketoconazole were inferior (0.194 and 0.183 microg/ml respectively) to that obtained with ketoconazole alone (0.390 microg/ml) . A synergistic effect is therefore obtained between ketoconazole on the one hand and two concentrations of Euphorbia characias latex.

Eur Arch Otorhinolaryngol, 2001 Aug, 258(6), 281 - 4
Advances and refinements in surgical voice rehabilitation after laryngectomy; Lichtenberger G; After Blom and Singer reported the construction of the so-called "duck bill" prosthesis in 1980, there have been quite a few newer voice prostheses constructed by other workers and new methods developed to predict the results, such as the insufflation and lidocaine test . Implanting the voice prosthesis with the Blom-Singer method has presented some problems and complications related to the puncture technique, therefore the following simplified esophagotracheal puncture technique is presented . The pharynx is opened with the laryngoscope which is then led up to the entrance of the esophagus . Through the laryngoscope, the distal end of the endo-extralaryngeal needle carrier, developed by the author and modified for mass production by R . Wolf Ltd., Germany, is led into the esophagus . The instrument is pushed forward as long as its distal bent, blunt end is palpable in the upper third of the tracheostoma . The needle with the thread (2/0 prolene) is pushed through from the inside, out in the upper third of the tracheostoma . A double wire forming a loop is led through the pointed metal cone (containing a built-in needle) and the catheter and tied behind a counterfixing pierced ball . The 2/0 prolene leading thread is then knotted with the wire . By pulling the thread and the wire, the pointed end of the metal cone with the needle built-in, perforates the soft parts and pulls the catheter with it (the same procedure will be used for primary puncture as well) . After this procedure the voice prosthesis can easily be placed in the fistula in a conventional manner . Using this technique, 59 patients could be implanted without puncture-related complications or problems . Problems, not related to the puncture technique, such as Candida albicans infection etc., were solved using the well-known treatment modalities . To stop leakage around the prosthesis, injection of Bioplastique into the soft tissue surrounding the fistula was used with success.

Clin Pediatr (Phila), 2001 Sep, 40(9), 503 - 6
Thrush in the breastfeeding dyad: results of a survey on diagnosis and treatment; Brent NB; Infection with Candida albicans in the breastfeeding dyad has been associated with extreme pain in the breastfeeding mother and may lead to premature weaning . There is presently a dearth of information on diagnosis, natural history, and treatment of this condition in the literature . Therefore, before such a trial was conducted, a survey was sent to experts in the field of lactation, the members of The Academy of Breastfeeding Medicine, on the diagnosis and treatment of thrush in the breastfeeding mother and baby . Results showed that the majority of respondents relied primarily on history and physical examination of the baby, but not the mother, to make the diagnosis . Laboratory tests were ordered only rarely . The most common initial treatment was oral nystatin for the infant and cream for the mother's breasts . This was followed by oral nystatin for the infant and oral fluconazole for the mother . Treatment of recurrence or persistence was again most commonly nystatin for both mother and infant, followed by oral nystatin for the infant and oral fluconazole for the mother or oral fluconazole for both . In the absence of controlled trials of this condition, these results may serve as suggestions for the clinician, until definitive data are available.

J Antimicrob Chemother, 2001 Oct, 48(4), 521 - 5
Breakthrough fungaemia in neonates and infants caused by Candida albicans and Candida parapsilosis susceptible to fluconazole in vitro; Krcmery V et al.; Breakthrough fungaemias due to Candida albicans and Candida parapsilosis appearing during fluconazole therapy in neonates and infants were assessed for risk factors and outcome . Forty fungaemias occurred during therapy with fluconazole within a 12 year national survey and were compared with 161 cases of non-breakthrough paediatric fungaemias . The agar disc diffusion test method was used for antifungal susceptibility testing and the Vitek system for species identification . Univariate and multivariate analysis for risk factors for breakthrough fungaemia were carried out . All the fungaemias were a result of strains susceptible to fluconazole at 0.25-4 mg/L in vitro {C . albicans (85%) and C . parapsilosis (15%)} . The mean number of positive blood cultures per episode was 2.2 . Sixteen children had 'early' breakthrough fungaemias (within 4-5 days) and 24 fungaemias appeared on day 6 and later . Mean fluconazole MICs in the 'early' group were 1.2, and 2.8 mg/L in the 'late' group (P < 0.03, t-test) . However, no difference was observed in the average dose of fluconazole used in the two groups . Neonatal age, total parenteral nutrition, very low birth weight, before surgery, central or umbilical venous catheterization and artificial ventilation were all significantly related to breakthrough fungaemia in univariate analysis but only central or umbilical venous catheterization were significant in multivariate analysis . The outcome of breakthrough fungaemia was better overall and attributable mortalities in non-breakthrough fungaemia was significantly higher in comparison with breakthrough fungaemia.

Invest Ophthalmol Vis Sci, 2001 Oct, 42(11), 2578 - 83
Molecular screening of donor corneas for fungi before excision; Kercher L et al.; PURPOSE: To develop panfungal and Candida albicans species-specific polymerase chain reaction (PCR) assays to screen donor eyes for fungal contamination before corneal excision . METHODS: PCR primers were designed for either the broad-spectrum detection of fungal DNA or the specific detection of C . albicans DNA . Their sequences were based on rDNA regions highly conserved among and specific to fungi and C . albicans, respectively . PCR conditions with the two primer sets were optimized and tested for sensitivity using purified C . albicans genomic DNA and a plasmid containing the relevant region of C . albicans DNA . The specificity of the primer sets was established using higher eukaryotic, fungal, prokaryotic, and viral DNAs as PCR templates . Donor eye swab specimens were collected before corneal excision . DNA was extracted from the specimens and tested by both PCR assays . RESULTS: The lower limit of detection for both primer sets was consistently 10(3) genome equivalents, when using genomic DNA as a template and 10(2) copies of plasmid . The fungal PCR assay amplified DNA from all fungal species tested but did not amplify any of the selected mammalian, bacterial, or viral DNA . The C . albicans PCR detected the C . albicans DNA but was negative for all other DNA substrates, including the other fungal templates . Thirty-five percent of the donor eye samples tested were positive for fungus, and 19% were positive for C . albicans DNA . CONCLUSIONS: The PCR assays allowed the rapid screening of DNA extracted from specimens collected from corneal donors for potential fungal contamination . The assay was highly sensitive and specific for screening corneal surfaces . The results suggest that approximately one-third of donor eyes tested harbor fungi on the ocular surface.

Mol Microbiol, 2001 Sep, 41(6), 1431 - 44
The KEX2 gene of Candida glabrata is required for cell surface integrity; Bader O et al.; Candida glabrata has emerged as one of the most common causes of candidosis . In order to identify factors that are necessary for viability and pathogenicity of this fungal pathogen, we analysed the role of the KEX2 gene, which codes for a regulatory endoproteinase that is known to process certain virulence factors in Candida albicans . The KEX2 gene from C . glabrata was cloned and found to have 51% and 62% identity and high structural similarities to the homologous counterparts in C . albicans and Saccharomyces cerevisiae . KEX2 was expressed at all time points investigated during growth in complex medium . In order to investigate the role of this putative regulatory proteinase, Kex2-deficient mutants were produced . In addition to known kex2 phenotypes, such as pH and calcium hypersensitivity, the mutants grew in cellular aggregates and were found to be hypersensitive to several antifungal drugs that target the cell membrane, including azoles, amorolfine and amphotericin B . Ultrastructural investigation after exposure to low doses of itraconazole showed azole-specific alterations such as enlarged vacuoles and proliferation of the cytoplasmatic membrane in the kex2 mutants, but not in the control strains . In contrast, antifungals such as 5-flucytosine and hydroxypyridones inhibited growth of the kex2 mutants and the control strains to the same extent . In an in vitro model of oral candidosis, kex2 mutants showed reduced tissue damage in the presence of itraconazole compared with the control infections . These data suggest that Kex2 is involved in the processing of proteins that are essential for cell surface integrity of C . glabrata.

J Clin Ultrasound, 2001 Sep, 29(7), 417 - 21
Fungal brain abscesses in neonates: Sonographic appearances and corresponding histopathologic findings; Marcinkowski M et al.; Extremely preterm neonates and neonates with predisposing conditions such as congenital or acquired immunodeficiency are at high risk for systemic fungal infection . Abscess formation in the brain is a severe complication that occurs in 70% of neonates with systemic fungal infection . Cerebral sonography can be used to diagnose abscesses in the brain in these patients . We report 2 sonographic presentations of fungal brain abscesses in neonates confirmed by postmortem histopathologic examination . The first patient, an extremely preterm neonate of 23 weeks' gestation with a systemic Candida albicans infection, had multiple small, round, hypoechoic lesions with echogenic rims in both brain hemispheres . The second patient, a term neonate with disseminated aspergillosis and DiGeorge syndrome, had a few large echogenic areas in the right periventricular region . Brain imaging should be considered in the diagnostic workup in neonates with suspected systemic fungal infection . Cerebral involvement can be diagnosed at the bedside with cerebral sonography .

Am J Kidney Dis . 2001 Oct;38(4):E20.
Linezolid for treatment of vancomycin-resistant enterococcal peritonitis; Bailey EM et al.; A 42-year-old woman on continuous ambulatory peritoneal dialysis (CAPD) developed peritonitis secondary to vancomycin-resistant Enterococcus faecium and Candida albicans while hospitalized for pneumonia . She was treated successfully with intravenous linezolid, fluconazole given by nasogastric tube, and removal of the peritoneal catheter . A concentration of linezolid above the minimum inhibitory concentration for most gram-positive pathogens, including vancomycin-resistant E faecium, was achieved in the dialysate fluid after an oral loading dose of 1200 mg . Additional data are needed to establish the role of linezolid in treating CAPD-associated peritonitis.

Phytochemistry, 2001 Oct, 58(4), 631 - 5
Quinones from Heliotropium ovalifolium; Guntern A et al.; Two new benzoquinones, heliotropinones A and B, have been isolated from the aerial parts of Heliotropium ovalifolium . Their structures were elucidated by spectrometric methods including high resolution electrospray ionization (ESI-HR), EI mass spectrometry, 1H, 13C and 2D NMR experiments . The two quinones demonstrated antifungal activities against Cladosporium cucumerinum and Candida albicans as well as antibacterial activity against Bacillus subtilis.

Phytochemistry, 2001 Oct, 58(4), 599 - 602
A weakly antimalarial biflavanone from Rhus retinorrhoea; Ahmed MS et al.; The biflavanone (2S,2"S)-7,7"-di-O-methyltetrahydroamentoflavone and five known flavonoids, 7-O-methylnaringenin, 7,3'-O-dimethylquercetin, 7-O-methylapigenin, 7-O-methylluteolin, and eriodictyol were isolated from the leaves of Rhus retinorrhoea Steud, Ex Olive . The biflavanone exhibited moderate antimalarial activity with IC50 0.98 microg/ml against Plasmodium falciparum (W2 Clone) and weak activity against P . falciparum (D6 Clone) with IC50 2.8 microg/ml . Nevertheless, it did not display any cytotoxicity . 7-O-Methylnaringenin showed weak antimicrobial activity against Candida albicans, C . krusei, Staphylococcus aureus, Mycobacterium smegmatis, M . intracellulare, and M . xenopi with MIC approximately 100 microg/ml . Characterization of each compound was based on spectral analysis and comparison with reported data.

Pediatr Dermatol, 2001 Jul-Aug, 18(4), 282 - 90
Effects of breathable disposable diapers: reduced prevalence of Candida and common diaper dermatitis; Akin F et al.; Infants wearing breathable disposable diapers experienced significantly less diaper dermatitis (DD) compared to infants wearing standard, nonbreathable disposable diapers in a series of double-blind clinical trials . Severe DD, including confirmed infection with Candida albicans, was reduced by 38-50% among infants wearing highly breathable (HB) diapers . The prevalence of DD was inversely related to the breathability of the garments . The inhibitory effect of breathable diapers on the survival of Candida was further confirmed in controlled experiments with adult volunteers . A suspension of C . albicans cells was applied to delineated sites on the volar forearm . Each site was then covered by a full-thickness patch from either an HB or a standard diaper . Survival of Candida colonies was reduced by almost two-thirds in the breathable diaper-covered sites compared to the control sites.

Nucleic Acids Res, 2001 Oct 1, 29(19), 4014 - 24
An active retrotransposon in Candida albicans; Holton NJ et al.; Tca2 is a Ty1/copia-type retrotransposon from the pathogenic yeast Candida albicans . It was originally identified as an abundant, linear, extrachromosomal, double-stranded DNA molecule . Here we show that Tca2 is widespread in C.albicans, but that the abundance of extrachromosomal Tca2 DNA varies greatly among different strains and is strongly dependent on the growth temperature . The relative levels of Tca2 RNA vary in a similar pattern to the extrachromosomal DNA, raising the possibility that the variations in extrachromosomal DNA levels are introduced predominantly at the level of transcription . We have also analysed the retrotranspositional activity of the element by developing a transposition assay involving a cloned Tca2 element tagged with a selectable marker gene that is activated by passage through an RNA intermediate . We show that the marked Tca2 is transpositionally active as transposed copies of the marked element became integrated at a variety of new positions in the genome and an intron in the donor element was precisely removed in the newly transposed copies . This is the first report of transposition in C.albicans.

J Clin Microbiol, 2001 Oct, 39(10), 3793 - 5
Performance of candida ID, a new chromogenic medium for presumptive identification of Candida species, in comparison to CHROMagar Candida; Willinger B et al.; Candida ID agar allows identification of Candida albicans and differentiation of other Candida species . In comparison with CHROMagar Candida, we evaluated the performance of this medium directly from 596 clinical specimens . In particular, detection of C . albicans after 24 h of incubation was easier on Candida ID (sensitivity, 96.8%) than on CHROMagar (sensitivity, 49.6%).

J Clin Microbiol, 2001 Oct, 39(10), 3491 - 4
Quantification of Candida albicans actin mRNA by the LightCycler system as a means of assessing viability in a model of cutaneous candidiasis; Okeke CN et al.; The LightCycler system (two-step reverse transcription-PCR-fluorescent hybridization {LC RT-PCR-FH}) was used to quantify Candida albicans actin mRNA as a means of assessing its viability in a reconstituted skin model of cutaneous candidiasis following the application of an antimycotic . A 192-bp ACT exon fragment was ligated into the pCR2.1 plasmid vector, and dilutions of the cloned insert (pACT; 4.092 kb) were used as the standard reference template . The LC RT-PCR-FH system could detect 1 fg of pACT, equivalent to 2.2 copies of the plasmid . The ACT exon-based PCR primers and FH probes were C . albicans specific, and electrophoretic analysis of the LC RT-PCR-FH assay product showed a 174-bp band in agarose gel . The number of copies of C . albicans ACT mRNA per milligram of tissue decreased with increasing amounts of amorolfine applied to a C . albicans-infected skin model, showing a reduction in viability . Detection and quantification of ACT mRNA in tissue by the LC RT-PCR-FH assay corresponded with cultural isolation of C . albicans from samples . The ACT mRNA-targeted LC RT-PCR-FH assay represents a sensitive, specific, rapid, and quantitative means of assessing the viability of C . albicans in infected tissue . This method may also be useful in evaluating the therapeutic efficacies of antifungal drugs in the treatment of various forms of candidiasis and other fungal diseases.

Gene, 2001 Sep 5, 275(1), 133 - 40
The Candida albicans gene encoding the cytoplasmic leucyl-tRNA synthetase: implications for the evolution of CUG codon reassignment; O'Sullivan JM et al.; In a number of Candida species the 'universal' leucine codon CUG is decoded as serine . To help understand the evolution of such a codon reassignment we have analyzed the Candida albicans leucyl-tRNA synthetase (CaLeuRS) gene (CaCDC60) . The predicted CaLeuRS sequence shows a significant level of amino acid identity to LeuRS from other organisms . A mitochondrial LeuRS (ScNAM2) homologue, which shared low identity with the CaLeuRS, was also identified in C . albicans . Antigenically-related LeuRSs were identified in a range of Candida species decoding the CUG codon as both serine and leucine, using an antibody raised against the N-terminal 15 amino acids of the CaLeuRS . Complementation experiments demonstrated that the CaLeuRS was able to functionally complement a Saccharomyces cerevisiae cdc60::kanMX null mutation . We conclude that there is no alteration in tRNA recognition and aminoacylation by the C . albicans LeuRS, which argues against it having a role in codon reassignment . The nucleotide sequences of the CaCDC60 and CaNAM2 genes were deposited at GenBank under Accession numbers AF293346 and AF352020, respectively.

Ann Pharmacother, 2001 Sep, 35(9), 1042 - 4
Transient hypoparathyroidism due to amphotericin B-induced hypomagnesemia in a patient with beta-thalassemia; Marcus N et al.; OBJECTIVE: To report a case of transient hypoparathyroidism that developed in a beta-thalassemic patient due to amphotericin B-induced hypomagnesemia . CASE SIJMMARY: A 21-year-old man with beta-thalassemia was treated with amphotericin B for Candida albicans intravenous line sepsis . After five days of treatment (cumulative dose 160 mg), he developed hypomagnesemia, which caused hypoparathyroidism and hypocalcemia; all three abnormalities resolved after the drug was withdrawn . DISCUSSIoN: Patients with beta-thalassemia may develop endocrinologic abnormalities due to excessive iron deposition . Some may have subclinical hypoparathyroidism that clinically emerges after even a mild homeostasis disturbance . Amphotericin B is associated with variable adverse effects including renal tubular insult, which may induce hypomagnesemia following relatively short treatment . The resolution of hypomagnesemia, hypocalcemia, and hypoparathyroidism in our patient after discontinuation of amphotericin B treatment suggests that the endocrine dysfunction was due to a drug-related adverse effect and not to parathyroid dysfunction caused by iron deposition . CONCLUSIONS: This case demonstrates a known but rarely reported adverse effect of amphotericin B, namely hypomagnesemia, that may occur even at a low cumulative dose . It also emphasizes that patients with an underlying disease, such as thalassemia, may be more susceptible to hypoparathyroidism and hypocalcemia during treatment with amphotericin B.

Yeast, 2001 Oct, 18(14), 1339 - 45
Cloning of a fatty acid synthase component FAS1 gene from Saccharomyces kluyveri and its functional complementation of S . cerevisiae fas1 mutant; Kajiwara S et al.; A gene encoding a fatty acid synthase component, FAS1, has been cloned from a genomic library of the polyunsaturated fatty acid (PUFA)-producing yeast Saccharomyces kluyveri . This gene (named Sk-FAS1) was found to contain an open reading frame of 6150 bp, coding for 2049 amino acids . The deduced Sk-FAS1 protein showed significant (75-59%) homology with FAS proteins from the other yeasts, including S . cerevisiae, Candida albicans and Yarrowia lipolytica . The substrate-binding sites of the acetyl transferase and malonyl/palmitoyl transferase domains, and the FMN- and NADPH-binding sites of the enoyl reductase domain, were all highly conserved . Expression of the Sk-FAS1 gene in S . cerevisiae complemented genetic disruption of the S . cerevisiae FAS1 gene (Sc-FAS1), suggesting the formation of a heterogeneous complex of Sk-FAS1 (beta) and Sc-FAS2 (alpha), which is able to function to synthesize fatty acids . Compared with the isogenic wild-type of S . cerevisiae, as well as S . kluyveri, the S . cerevisiae fas1 mutant carrying the Sk-FAS1 gene showed an increase in the relative amount of 16-carbon fatty acids and a decrease in 18-carbon fatty acids .

Bone Marrow Transplant, 2001 Aug, 28 Suppl 1, S18 - 21
Infections after stem cell transplantation in children: state of the art and recommendations; Dini G et al.; At the workshop on infections after stem cell transplantation (SCT) in children, the following topics were introduced by invited speakers and discussed with the audience: empirical antimicrobial therapy in the pre-engraftment period, early diagnosis of fungal and viral infections and possibilities to treat them and the possible role of G-CSF early post-SCT . Episodes of fever in the pre-engraftment period mostly are unexplained, and in about one quarter due to bacteremia, mostly by Gram-positive cocci . No single drug or combination of drugs used for antimicrobial therapy is superior, neither does it cover 100% of the pathogens . Close microbiological surveillance of the patients and knowledge of the local microbial epidemiology are requested for optimal therapy . Early fungal infections are reactivations of pre-SCT infections, late fungal infections mostly are associated with failure of engraftment or GvHD and its treatment . Except for suggestive ultrasound or CT-scan abnormalities, the possibilities for early diagnosis are limited c.q . not reliable . Fluconazol prophylaxis is recommended to prevent Candida albicans invasion . A number of new antifungal drugs are being tested in phase I and II studies . CMV, EBV and adenoviruses may reactivate after SCT, causing severe disease with a high mortality, especially in non-HLA-identical donor-recipient combinations . Frequent surveillance cultures for CMV and adenoviruses, pp65-CMV antigen detection in WBC and PCR techniques for CMV, EBV and adenoviruses all have their own contribution to the early diagnosis of dissemination of the viral infection . Therapeutical possibilities, except with respect to ganciclovir and foscarnet for CMV infection, are still limited . The effectiveness of cidofovir is under study . Adoptive therapy with virus-specific CTL's probably represents the new frontier . G-CSF administration early after SCT has a beneficial effect on PMN recovery, hospitalization time, use of antibiotics and total parenteral nutrition requirement in children undergoing allogeneic and autologous BMT . No benefit is observed in children undergoing peripheral blood SCT . The routine use of G-CSF in the latter group of patients is not justified.

Curr Opin Crit Care, 2001 Aug, 7(4), 238 - 41
Optimal use of existing and new antifungal drugs; Rogers TR; Clinicians are increasingly aware that fungal pathogens are a significant cause of morbidity and mortality in hospitalized patients . Historically, these infections occurred in severely immunocompromised patients who were undergoing treatment for hematological malignancy or solid organ transplantation . Currently, however, systemic fungal infections are commonly seen in debilitated patients who are being nursed in intensive care or high-dependency units . These infections are mostly caused by Candida albicans but there is a growing proportion of strains of non- albicans Candida spp, some with reduced susceptibility to commonly used antifungals . The limited armamentarium of antifungal agents to date has meant that amphotericin B continues to be considered the most effective therapeutic agent albeit with a poor record of treatment-limiting side effects . The past decade has seen some encouraging developments in antifungal therapy . Three lipid formulations of amphotericin B showing reduced toxicity compared with the desoxycholate formulation are now licensed . There are three investigational triazoles currently undergoing evaluation that should prove important additions to existing members of this class . The echinocandin caspofungin is the first of a new class of antifungal agents with a novel mode of action, which has recently been approved for use in the United States.

Am J Med Sci, 2001 Sep, 322(3), 160 - 2
Implantable cardioverter-defibrillator endocarditis secondary to Candida albicans; Brown LA et al.; The implantable cardioverter-defibrillator (ICD) represents an important advance in the treatment of ventricular arrhythmias, but infection has remained a serious complication of device implantation . Fungal infections associated with these devices are uncommon, with only 4 cases previously reported . We describe a case of ICD-associated endocarditis caused by Candida albicans that was successfully treated with complete device explantation and prolonged antifungal therapy, and we review the features of ICD-related fungal infections.

J Biosci, 2001 Sep, 26(3), 333 - 9
Specificity of drug transport mediated by CaMDR1: a major facilitator of Candida albicans; Kohli A et al.; CaMDR1 encodes a major facilitator superfamily (MFS) protein in Candida albicans whose expression has been linked to azole resistance and which is frequently encountered in this human pathogenic yeast . In this report we have overexpressed CaMdr1p in Sf9 insect cells and demonstrated for the first time that it can mediate methotrexate (MTX) and fluconazole (FLC) transport . MTX appeared to be a better substrate for CaMdr1p among these two tested drugs . Due to severe toxicity of these drugs to insect cells, further characterization of CaMdr1p as a drug transporter could not be done with this system . Therefore, as an alternative, CaMdr1p and Cdr1p, which is an ABC protein (ATP binding cassette) also involved in azole resistance in C . albicans, were independently expressed in a common hypersensitive host JG436 of Saccharomyces cerevisiae . This allowed a better comparison between the functionality of the two export pumps . We observed that while both FLC and MTX are effluxed by CaMdr1p, MTX appeared to be a poor substrate for Cdr1p . JG436 cells expressing Cdr1p thus conferred resistance to other antifungal drugs but remained hypersensitive to MTX . Since MTX is preferentially transported by CaMdr1p, it can be used for studying the function of this MFS protein.

J Inorg Biochem, 2001 Sep, 86(2-3), 565 - 71
Synthesis, structure and biological activity of nickel(II) complexes of 5-methyl 2-furfural thiosemicarbazone; Jouad EM et al.; 5-Methyl 2-furfuraldehyde thiosemicarbazone (M5HFTSC) with nickel(II) leads to three types of complexes: {Ni(M5HFTSC)(2)X(2)}, {Ni(M5FTSC)(2)} and {Ni(M5FTSC)(2)} x 2DMF . In the first type the ligand remains in thione form, while in the two other, the anionic thiolato form is involved . The species {Ni(M5HFTSC)(2)X(2)} has been characterized spectroscopically . The structures of {Ni(M5FTSC)(2)} x 2DMF and {Ni(M5FTSC)(2)} have been solved using X-ray diffraction . Biological studies of {Ni(M5HFTSC)(2)Cl(2)} have been carried out in vitro for antifungal activity on human pathogenic fungi, Aspergillus fumigatus and Candida albicans, and in vivo for toxicity on mice . The results are compared to those of the ligand, the metal salt and a similar copper complex {Cu(M5HFTSC)Cl(2)}.

Biochem J, 2001 Oct 1, 359(Pt 1), 35 - 45
Characterization of unique amphipathic antimicrobial peptides from venom of the scorpion Pandinus imperator; Corzo G et al.; Two novel antimicrobial peptides have been identified and characterized from venom of the African scorpion Pandinus imperator . The peptides, designated pandinin 1 and 2, are alpha-helical polycationic peptides, with pandinin 1 belonging to the group of antibacterial peptides previously described from scorpions, frogs and insects, and pandinin 2 to the group of short magainin-type helical peptides from frogs . Both peptides demonstrated high antimicrobial activity against a range of Gram-positive bacteria (2.4-5.2 microM), but were less active against Gram-negative bacteria (2.4-38.2 microM), and only pandinin 2 affected the yeast Candida albicans . Pandinin 2 also demonstrated strong haemolytic activity (11.1-44.5 microM) against sheep erythrocytes, in contrast with pandinin 1, which was not haemolytic . CD studies and a high-resolution structure of pandinin 2 determined by NMR, showed that the two peptides are both essentially helical, but differ in their overall structure . Pandinin 2 is composed of a single alpha-helix with a predominantly hydrophobic N-terminal sequence, whereas pandinin 1 consists of two distinct alpha-helices separated by a coil region of higher flexibility . This is the first report of magainin-type polycationic antimicrobial peptides in scorpion venom . Their presence brings new insights into the mode of action of scorpion venom and also opens new avenues for the discovery of novel antibiotic molecules from arthropod venoms.

Int J Fertil Womens Med, 2001 Jul-Aug, 46(4), 210 - 4
Studies on the significance of positive bacterial semen cultures in male fertility in Nigeria; Onemu SO et al.; OBJECTIVE: Early warning signals (EWS) of altered reproductive potential may be very important in the prevention and management of male infertility . The presence of bacteria in semen (bacterisemia) may be an EWS . This was evaluated by determining the incidence of bacteria in semen of males with fertility problems in Benin City by culturing their semen . METHODS: Diluted semen samples were cultured on blood agar, chocolate agar, MacConkey agar, nutrient agar, and sabouraud dextrose agar slants for the isolation of micro-organisms . Colonies of a single type of micro-organism (>10(3) cfu/mL) were picked for identification and sensitivity tests using antibacterial agents . Each semen sample was further processed for spermatozoal morphology and motility, presence of peroxidase-positive cells, and other accompanying cells . Correlative studies on the relationship between bacterisemia and semen/spermatozoa variables such as total number and motility were also done . RESULTS: Pathogenic micro-organisms were present in 78/163 (47.1%) semen samples . The microbial isolates were Staphylococcus aureus 35 (43.7%), Klebsiella species 22 (28.2%), Escherichia coli 9 (11.5%), and Candida albicans 6 (7.7%) . The bacterial isolates were most sensitive to ceftazidime and pefloxacin, and least to amoxycillin and tetracycline . There was a positive correlation (r = .9774) between azoospermia in males and presence of Candida albicans in semen, as well as between the presence of micro-organisms and poor semen quality (r = .8563), and the presence of micro-organisms and reduced motility (r = .8246) . CONCLUSION: Presence of pathogenic micro-organisms in semen, which may be related to a breach in the integrity of the blood-testes barrier, may provide early warning signals of impairment of male fertility.

J Biol Chem, 2001 Nov 16, 276(46), 43049 - 55 Epub 2001 Sep 18.
Identification of a Candida albicans ferrichrome transporter and its characterization by expression in Saccharomyces cerevisiae; Ardon O et al.; Saccharomyces cerevisiae can accumulate iron through the uptake of siderophore-iron . Siderophore-iron uptake can occur through the reduction of the complex and the subsequent uptake of iron by the high affinity iron transporter Fet3p/Ftr1p . Alternatively, specific siderophore transporters can take up the siderophore-iron complex . The pathogenic fungus Candida albicans can also take up siderophore-iron . Here we identify a C . albicans siderophore transporter, CaArn1p, and characterize its activity . CaARN1 is transcriptionally regulated in response to iron . Through expression studies in S . cerevisiae strains lacking endogenous siderophore transporters, we demonstrate that CaArn1p specifically mediates the uptake of ferrichrome-iron . Iron-ferrichrome and gallium-ferrichrome, but not desferri-ferrichrome, could competitively inhibit the uptake of iron from ferrichrome . Uptake of siderophore-iron resulting from expression of CaARN1 under the control of the MET25-promoter in S . cerevisiae was independent of the iron status of the cells and of Aft1p, the iron-sensing transcription factor . These studies demonstrate that the expression of CaArn1p is both necessary and sufficient for the nonreductive uptake of ferrichrome-iron and suggests that the transporter may be the only required component of the siderophore uptake system that is regulated by iron and Aft1p.

J Biol Chem, 2001 Nov 23, 276(47), 43784 - 91 Epub 2001 Sep 18.
Candida albicans expresses an unusual cytoplasmic manganese-containing superoxide dismutase (SOD3 gene product) upon the entry and during the stationary phase; Lamarre C et al.; We report here that in addition to a cytoplasmic copper-zinc-containing superoxide dismutase (SOD) and a mitochondrial manganese-containing SOD, Candida albicans expresses a third SOD gene (SOD3) . The deduced amino acid sequence contains all of the motifs found in previously characterized manganese-containing SODs, except the presence of a mitochondrial transit peptide . Recombinant Sod3p expressed and purified from Escherichia coli is a homotetramer with a subunit mass of 25.4 kDa . Mass absorption spectrometry detected the presence of both iron and manganese in purified Sod3p but, as determined by metal replacement experiments, the enzyme displays activity only when bound to manganese . Overexpression of SOD3 was shown to rescue the hypersensitivity to redox cycling agents of a Saccharomyces cerevisiae mutant lacking the cytoplasmic copper-zinc-containing SOD . Northern blot analyses showed that the transcription of SOD3 is induced neither by the transition from the yeast to the mycelial form of C . albicans nor by drug-induced oxidative stress . In continuous cultures, the expression of SOD3 was strongly stimulated upon the entry and during the stationary phase, concomitantly with the repression of SOD1 . We conclude that Sod3p is an atypical cytosolic manganese-containing superoxide dismutase that is involved in the protection of C . albicans against reactive oxygen species during the stationary phase.

Microb Pathog, 2001 Oct, 31(4), 159 - 72
Involvement of alpha(v)beta3 integrin-like receptor and glycosaminoglycans in Candida albicans germ tube adhesion to vitronectin and to a human endothelial cell line; Santoni G et al.; The present study was undertaken to investigate the expression of alpha(v)beta3 and alpha(v)beta5 integrin-like vitronectin receptors (VNRs) on Candida albicans germ tube and their involvement in its adhesion to vitronectin (VN) and human endothelial cells . By immunofluorescence and FACS analysis, several monoclonal antibodies directed against human alpha(v) or beta3 integrin subunit or alpha(v)beta3 and alpha(v)beta5 heterodimers, positively stained C . albicans germ tubes . C . albicans germ tubes specifically adhered (45-50%) to VN and this adhesion was markedly inhibited by RGD-, but not RGE-containing peptides . Adhesion of C . albicans germ tubes to VN was strongly inhibited by anti-alphav, anti-beta3 or anti-alpha(v)beta3, but not by alpha(v)beta5 monoclonal antibody . C . albicans germ tube adhesion to VN was also inhibited by glycosaminoglycans (GAGs) such as heparin or chondroitin sulphate . Finally, we show that C . albicans germ tubes adhere to the human EA.hy 926 endothelial cell line . This adhesion is markedly blocked by anti-beta3 monoclonal antibody, GRGDSP peptide or heparin, and is completely abolished by their combination . Overall these results indicate that C . albicans germ tube adherence to VN and to a human endothelial cell line is mediated by alpha(v)beta3, but not by alpha(v)beta5-like integrin, and depends on GAGs which may act by regulating alpha(v)beta3 integrin-like/VN adhesive interaction .

Int Immunopharmacol, 2001 Sep, 1(9-10), 1797 - 811
Protective effects of a protein-bound polysaccharide, PSK, on Candida albicans infection in mice via tumor necrosis factor-alpha induction; Ohmura Y et al.; We investigated the protective mechanism of a protein-bound polysaccharide, PSK, against lethal infection with Candida albicans (C . albicans) in mice . (1) In BALB/c mice inoculated intravenously with C . albicans, the intraperitoneal (ip) administration of PSK increased survival rates and prolonged the survival period depending on the time of administration, the dosage, and the size of fungal inoculum; the maximal effect was obtained when PSK 250 mg/kg was ip administered to mice 24 h before inoculation of 1 x 10(6) C . albicans (30 days survivors showed 60% and the mean survival period of mice with fatal infection increased 209%) . (2) The protective effect of PSK was significantly decreased in mice treated with cyclophosphamide or carrageenan, or in mice treated previously with anti-tumor necrosis factor-alpha (TNF-alpha) antibody . (3) The administration of PSK significantly enhanced the expression of TNF-alpha gene in spleen and increased leukocyte functions from 6 h to 1 day after inoculation . (4) When the PSK fraction subjected to hydrolysis with beta1-3 glucanase or hydrazine was used instead of PSK, the anti-fungal activities were significantly decreased . These findings suggested that the protective effect of PSK on lethal C . albicans infection in mice was mainly produced via TNF-alpha functions, and that beta 1-3 glucan and protein moiety in PSK molecule were involved in the expression of the activities.

Yeast, 2001 Sep 30, 18(13), 1217 - 25
Functional isolation of the Candida albicans FCR3 gene encoding a bZip transcription factor homologous to Saccharomyces cerevisiae Yap3p; Yang X et al.; We have isolated a C . albicans gene, named FCR3 (for fluconazole resistance 3), based upon its ability to suppress the FCZ hypersusceptibility of a Saccharomyces cerevisiae mutant strain (JY312) lacking the transcription factors Pdr1p and Pdr3p . The FCR3 ORF (1200 bp) encodes a 399 amino acid protein containing a basic leucine zipper (bZip) domain . Fcr3p displays the highest level of sequence homology with the S . cerevisiae Yap3p protein (34% identity, 45% similarity) . We had previously shown that deletion of the PDR5 gene encoding a multidrug transporter completely abolished the ability of FCR3 to suppress the FCZ hypersusceptibility of JY312, suggesting that FCR3 confers FCZ resistance by activating PDR5 expression . We show here that the beta-galactosidase activity of a PDR5 promoter-lacZ construct in JY312 is increased two-fold upon FCR3 overexpression, demonstrating that FCR3 regulates PDR5 at the transcriptional level . We also show that FCR3 overexpression not only suppresses the hypersusceptibility of JY312 to 4-nitroquinoline-N-oxide (4-NQO) but also confers higher levels of resistance to this compound as compared to the wild-type KY320 strain . Since PDR5 is not involved in 4-NQO resistance, this result indicates that FCR3 can also activate the transcription of other genes that can confer 4-NQO resistance . Finally, Northern blot analysis indicates that FCR3 encodes a single 2.4 kb RNA transcript in C . albicans, suggesting that the FCR3 mRNA contains long 5' and/or 3' untranslated regions . The nucleotide sequence of the FCR3 gene has been deposited at GenBank under Accession No . AF342983 .

Chemotherapy, 2001 Sep-Oct, 47(5), 350 - 3
Effects of cefepime and meropenem on the gastrointestinal colonization of surgical patients by Candida albicans; Samonis G et al.; BACKGROUND: The study evaluated the effects of cefepime and meropenem on the gastrointestinal (GI) colonization of surgical patients by Candida albicans . PATIENTS AND METHODS: Twenty adult surgical patients who received intravenously either of these antibiotics as monotherapy for the treatment of an existing infection were studied prospectively . Ten patients received cefepime (2.0 g twice a day), and another ten meropenem (1.0 g every 8 h) for 7 days . Quantitative stool cultures for C . albicans were performed immediately before, at the end, and 1 week after the end of antibiotic treatment . RESULTS: Both antibiotics increased the GI colonization of patients by Candida . Meropenem caused a higher increase (2.0 log(10) CFU/g of stool) as compared to cefepime (1.7 log(10) CFU/g of stool) . However, these increases were statistically not significant . CONCLUSION: Cefepime and meropenem when given to sensitive patients do not increase significantly the risk of Candida infection originating in the GI tract .

J Biol Chem, 2001 Nov 23, 276(47), 43767 - 74 Epub 2001 Sep 17.
A helix-loop-helix peptide at the upper lip of the active site cleft of lysozyme confers potent antimicrobial activity with membrane permeabilization action; Ibrahim HR et al.; Recently, we have found that partially unfolded lysozyme exerts broad spectrum antimicrobial action in vitro against Gram-negative and Gram-positive bacteria independent of its catalytic activity . In parallel, an internal peptide (residues 98-112) of hen egg white lysozyme, obtained after digestion with clostripain, possessed broad spectrum antimicrobial action in vitro . This internal peptide is part of a helix-loop-helix domain (87-114 sequence of hen lysozyme) located at the upper lip of the active site cleft of lysozyme . The helix-loop-helix (HLH) structures are known motifs commonly found in membrane-active and DNA-binding proteins . To evaluate the contribution of the HLH peptide to the antimicrobial properties of lysozyme, the HLH sequence and its secondary structure derivatives of chicken and human lysozyme were synthesized and tested for antimicrobial activity against several bacterial strains . We found that the full HLH peptide of both chicken and human lysozymes was potently microbicidal against both Gram-positive and Gram-negative bacteria and the fungus Candida albicans . The N-terminal helix of HLH was specifically bactericidal to Gram-positive bacteria, whereas the C-terminal helix was bactericidal to all tested strains . Outer and inner membrane permeabilization studies, as well as measurements of transmembrane electrochemical potentials, provided evidence that HLH peptide and its C-terminal helix domain kill Gram-negative bacteria by crossing the outer membrane via self-promoted uptake and causing damage to the inner membrane through channel formation . The results are discussed in terms of proposed mechanisms for the catalytically independent antimicrobial activity of lysozyme that offer a new strategy for the design of potential antimicrobial drugs in the treatment of infectious diseases.

AIDS Res Hum Retroviruses, 2001 Sep 1, 17(13), 1273 - 7
Delayed hypersensitivity skin testing to mumps and Candida albicans antigens is normal in middle-aged HTLV-I- and-II-infected U.S . cohorts; Murphy EL et al.; It has been reported that human T cell lymphotropic virus (HTLV)-I-infected persons in Japan have decreased delayed hypersensitivity skin test reactivity to tuberculin purified protein derivative (PPD), but HTLV-I- or -II-infected persons do not generally develop opportunistic infections . We administered standardized intradermal testing with PPD, mumps, and Candida albicans antigens to 31 HTLV-I, 48 HTLV-II, and 143 seronegative subjects in the United States . Reactivity at 48 hr was compared among the three groups . Response rates to PPD were very low in all subjects . Fifty-five percent of seronegative subjects did not react to mumps antigen, compared with 55% of HTLV-I {adjusted odds ratio (OR) = 0.79, 95% confidence interval (CI) 0.27-2.33} and 38% of HTLV-II (OR = 0.73, 95% CI 0.33-1.64) . Fifty-one percent of seronegatives did not react to Candida albicans antigen, compared with 34% of HTLV-I (OR = 0.37, 95% CI 0.15-0.93) and 46% of HTLV-II (OR = 0.71, 95% CI 0.34-1.52) . Anergy was present in 33% of seronegatives, 28% of HTLV-I (OR = 0.60, 95% CI 0.20-1.78), and 19% of HTLV-II (OR = 0.56, 95% CI 0.22-1.44) . HTLV-I- and -II-infected persons appear to have intact delayed hypersensitivity skin test responses to mumps and Candida albicans antigens.

Eur J Biochem, 2001 Sep, 268(18), 4833 - 41
Protein-bound conformation of a specific inhibitor against Candida albicans myristoyl-CoA:protein N-myristoyltransferase in the ternary complex with CaNmt and myristoyl-CoA by transferred NOE measurements; Miura T et al.; Transferred nuclear Overhauser enhancement (trNOE) experiments have been performed to study the bioactive conformation(s) of Ro09-3472/000 derivatives in the ternary complex with Candida albicans myristoyl-CoA: protein N-myristoyltransferase (CaNmt) and myristoylCoA (MyrCoA) . A critical step in the trNOE study is to identify 'true' trNOEs in the spectra . Nonspecific binding of ligands to target proteins and/or spin diffusion effects can give rise to 'false' trNOEs, which may lead to an incorrect conclusion if used to derive bound conformations . In this study for all ligands the observed trNOEs arose from specific binding interactions with the active site of CaNmt . This was shown by displacing the ligand with the known tightly binding active-site inhibitor 1 {Devadas, B., Zupec, M.E., Freeman, S.K., Brown, D.L., Nagarajan, S., Sikorski, J.A., McWherter, C.A., Getman, D . P . & Gordon, J.I . (1995) J . Med . Chem . 38, 1837-1840} and measuring the resonance linewidths in the NMR spectrum before and after addition of the competitive inhibitor . The compounds were also tested for nonspecific protein binding with bovine serum albumin (BSA) using the same METHOD: Of the six compounds tested, Ro09-3700/001 (racemate) and its optically pure enantiomers, Ro09-4764/001(S) and Ro09-4765/001(R), showed both specific binding to CaNmt and no interaction with BSA . The NMR data of these molecules in the ternary complex with CaNmt/MyrCoA could thus be used for a detailed structural analysis . Thereby, the conformation of the bound ligand was obtained from a conformational search using the observed trNOEs as a selection filter . The NMR-determined conformations are in good agreement with the recently solved CaNmt-bound X-ray structures of two similar Ro09-3472/000 derivatives.

Antimicrob Agents Chemother, 2001 Oct, 45(10), 2787 - 92
Animal pharmacokinetics and interspecies scaling of sordarin derivatives following intravenous administration; Aviles P et al.; Sordarin derivatives constitute a new group of synthetic antifungal agents that selectively inhibit fungal protein synthesis . They have demonstrated in vitro activity against the most important fungal pathogens, both yeast and filamentous . This new family of compounds has also shown in vivo activity against murine Candida albicans, Histoplasma capsulatum, and Coccidioides immitis experimental infections, as well as against Pneumocystis carinii pneumonia in rats . After intravenous dosing in animals, both the area under the concentration-time curve and the elimination half-life were highest in Cynomolgus monkeys, followed by those in rats, mice, and rabbits . The volume of distribution at steady state for sordarin derivatives was similar in all species tested . The clearance in rats and mice was higher than for other species . GM 237354, a sordarin derivative, was characterized by high serum protein binding in mouse, rat, and monkey serum (unbound fraction, < or =5%) . An indirect evaluation of the effect of liver function upon the metabolism of this class of compounds has been made in animals with impaired liver function such as Gunn rats, as well as in allometric studies that showed better correlations of half-life to liver blood flow than to animal body weight . Linearity of the main pharmacokinetic parameters was demonstrated after intravenous dosing of the representative compound GM 193663 at 10 and 20 mg/kg of body weight in rats . Allometry was used to determine whether human pharmacokinetic parameters can be predicted from animal data by regression analysis against body weight and liver blood flow . All these results have demonstrated that the human pharmacokinetics of sordarin derivatives can be forecast from animal data.

Antimicrob Agents Chemother, 2001 Oct, 45(10), 2746 - 54
Correlation between in vitro and in vivo activities of GM 237354, a new sordarin derivative, against Candida albicans in an in vitro pharmacokinetic-pharmacodynamic model and influence of protein binding; Aviles P et al.; The antifungal effect of GM 237354, a sordarin derivative, was studied in an in vitro pharmacokinetic (PK)-pharmacodynamic dynamic system (bioreactor) which reproduces PK profiles observed in a previously described model of drug efficacy against murine systemic candidiasis . Immunocompetent mice infected intravenously with 10(5) CFU of Candida albicans were treated with GM 237354 at 2.5, 10, and 40 mg/kg of body weight every 8 h subcutaneously for 7 days . Free concentrations in serum were calculated by multiplying total concentrations measured in vivo by 0.05, the free fraction determined in vitro by equilibrium dialysis . In the bioreactor the inoculum was approximately 10(6) CFU/ml; and a one-compartment PK model was used to reproduce the PK profiles of free and total GM 237354 in serum obtained in mice, and clearance of C . albicans was measured over 48 h . A good correlation was observed when the in vivo fungal kidney burden and the area under the survival time curve were compared with the in vitro broth "burden," although only when free in vivo levels in serum were reproduced in vitro . GM 237354 displayed a 3-log decrease effect both in vivo and in vitro . The very few reports available on in vitro-in vivo correlations have been obtained with antibiotics . The good in vitro-in vivo correlation obtained with an antifungal agent shows that the in vitro dynamic system could constitute a powerful investigational tool prior to assessment of the efficacy of an anti-infective agent in animals and humans.

Antimicrob Agents Chemother, 2001 Oct, 45(10), 2676 - 84
Prevalence of molecular mechanisms of resistance to azole antifungal agents in Candida albicans strains displaying high-level fluconazole resistance isolated from human immunodeficiency virus-infected patients; Perea S et al.; Molecular mechanisms of azole resistance in Candida albicans, including alterations in the target enzyme and increased efflux of drug, have been described, but the epidemiology of the resistance mechanisms has not been established . We have investigated the molecular mechanisms of resistance to azoles in C . albicans strains displaying high-level fluconazole resistance (MICs, > or =64 microg/ml) isolated from human immunodeficiency virus (HIV)-infected patients with oropharyngeal candidiasis . The levels of expression of genes encoding lanosterol 14alpha-demethylase (ERG11) and efflux transporters (MDR1 and CDR) implicated in azole resistance were monitored in matched sets of susceptible and resistant isolates . In addition, ERG11 genes were amplified by PCR, and their nucleotide sequences were determined in order to detect point mutations with a possible effect in the affinity for azoles . The analysis confirmed the multifactorial nature of azole resistance and the prevalence of these mechanisms of resistance in C . albicans clinical isolates exhibiting frank fluconazole resistance, with a predominance of overexpression of genes encoding efflux pumps, detected in 85% of all resistant isolates, being found . Alterations in the target enzyme, including functional amino acid substitutions and overexpression of the gene that encodes the enzyme, were detected in 65 and 35% of the isolates, respectively . Overall, multiple mechanisms of resistance were combined in 75% of the isolates displaying high-level fluconazole resistance . These results may help in the development of new strategies to overcome the problem of resistance as well as new treatments for this condition.

J Oral Rehabil, 2001 Aug, 28(8), 755 - 65
Candida albicans growth on thermal cycled materials for maxillofacial prostheses in vitro; Nikawa H et al.; In the present study, the growth of a single isolate of Candida albicans on saliva-, serum-coated or protein free (uncoated), thermocycled (4-70 degrees C for 1 min, respectively; 0, 1000 and 10 000 times) 15 commercial maxillofacial materials was investigated, by monitoring pH changes in growth media . The inhibitory effect of the tissue conditioners on fungal growth was analysed using three parameters viz: (i) delay in the onset of the rapid decline in pH (ii) reduction in the rate of pH change and (iii) the pH minima reached . In the case of control materials (non-thermocycled and uncoated), significant antifungal effect was observed with two products . However, the antifungal effect of the materials was significantly reduced both by thermal cycling (Analysis of covariance {ANOVA}; P < 0.01) and a layer of protein coating (saliva, P < 0.05; serum, P < 0.01) . When the interrelation between three parameters of fungal growth and the surface hydrophobicity of the materials were analysed, minimum pH of fungal growth on 10 000-thermocycled materials correlated well with the contact angles of the materials (Student t-test, P < 0.01), suggesting that thermocycling process reduced the unpolymerized components of the materials which showed the antifungal effects, resulted in that the cell growth depends on the surface hydrophobicity of the specimens . These results, taken together, suggest that the ageing of the materials and the biological fluids of the host enhanced the fungal growth on maxillofacial materials.

Med Mycol, 2001 Aug, 39(4), 341 - 52
Molecular epidemiology of Candida albicans strains isolated from the oropharynx of HIV-positive patients at successive clinic visits; Lasker BA et al.; Candida albicans strain diversity and fluconazole resistance were prospectively analyzed in oral strains from 29 adult human immunodeficiency virus (HIV)-positive patients followed for > 1 year who had five or more culture-positive clinic visits . Molecular typing consisted of genomic blots probed with the Ca3 repetitive element . Sixteen patients had one or more episodes of oropharyngeal candidiasis (OPC), 12 (75%) maintained the original genotype, whereas the remaining four patients had a succession of 2-3 genotypes . The original genotype, either alone or mixed with another strain or with non-C . albicans Candida spp., was recovered from oral lesions in 13 of 15 evaluable (86.7%) patients . C . dubliniensis was the infecting yeast in the remaining two patients . Different patterns of fluconazole resistance occurred in three OPC patients . One patient's infecting strain became less susceptible . A second patient was infected with a resistant genotype and a progressively more susceptible minor genotype variant . C . dubliniensis isolates from the third patient varied in susceptibility . Thirteen colonized patients who never developed OPC harbored a greater variety of C . albicans genotypes (2-6) than their infected counterparts (P = 0.35) . OPC patients maintained their original endogenous C . albicans strains for prolonged periods, whether or not they demonstrated decreased in vitro susceptibility to fluconazole . The adaptation and maintenance of an endogenous C . albicans strain within its host may be linked to as yet uncharacterized factors.

Med Mycol, 2001 Aug, 39(4), 303 - 13
Aspartyl proteinases of Candida albicans and their role in pathogenicity; De Bernardis F et al.; Among the putative virulence factors of Candida albicans, secreted aspartic proteinases (Sap, encoded by a family of at least nine genes) continue to attract the attention of many investigators studying the pathogenesis of candidiasis . Several early studies documented a correlation between the levels of Sap secretion and the virulence of different strains, but much stronger support for this role has been provided by more recent data on differential SAP gene(s) expression in ex vivo and in vivo models, the outcome of infections with SAP-deleted mutants, and use of Sap immunogens . In particular, some SAP-deleted strains suffered a substantial loss of virulence, and, more interestingly, this was specifically associated with selected gene products and selected experimental pathologies . Moreover, anti-Sap antibodies have been shown to mediate a degree of protection in an experimental, mucosal candidiasis model . There is now initial evidence that distinct Saps are differentially produced in various Candida illnesses or stages of them . The exact mechanisms of each Sap involvement in any particular Candida disease, with special regard to human infections, and how the immune system deals with Sap, are critical issues for future research . An answer to these questions will possibly facilitate the generation of Sap-based anticandidal drugs or immunotherapeutics.

Infect Immun, 2001 Oct, 69(10), 6110 - 8
T cells augment monocyte and neutrophil function in host resistance against oropharyngeal candidiasis; Farah CS et al.; The purpose of this study was to identify the cell populations involved in recovery from oral infections with Candida albicans . Monoclonal antibodies specific for CD4+ cells, CD8+ cells, and polymorphonuclear leukocytes were used to deplete BALB/c and CBA/CaH mice of the relevant cell populations in systemic circulation . Monocytes were inactivated with the cytotoxic chemical carrageenan . Mice were infected with 10(8) C . albicans yeast cells and monitored for 21 days . Systemic depletion of CD4+ and CD8+ T lymphocytes alone did not increase the severity of oral infection compared to that of controls . Oral colonization persisted in animals treated with head and neck irradiation and depleted of CD4+ T cells, whereas infections in animals that received head and neck irradiation alone or irradiation and anti-CD8 antibody cleared the infection in a comparable fashion . The depletion of polymorphonuclear cells and the cytotoxic inactivation of mononuclear phagocytes significantly increased the severity of oral infection in both BALB/c and CBA/CaH mice . High levels of interleukin 12 (IL-12) and gamma interferon (IFN-gamma) were produced by lymphocytes from the draining lymph nodes of recovering animals, whereas IL-6, tumor necrosis factor alpha, and IFN-gamma were detected in the oral mucosae of both naive and infected mice . The results indicate that recovery from oropharyngeal candidiasis in this model is dependent on CD4+-T-cell augmentation of monocyte and neutrophil functions exerted by Th1-type cytokines such as IL-12 and IFN-gamma.

Eur J Anaesthesiol, 2001 Oct, 18(10), 687 - 94
Antimicrobial activity of ropivacaine and other local anaesthetics; Aydin ON et al.; BACKGROUND AND OBJECTIVE: It is claimed that local anaesthetics have antimicrobial properties . Our aim was to investigate the antimicrobial effects of different concentrations of ropivacaine, bupivacaine, lidocaine and prilocaine on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans . METHODS: All local anaesthetic dilutions were exposed to microorganisms for 0, 30, 60, 120, 240 min at room temperature . The inoculums taken from diluted suspensions were reinoculated on blood agar and incubated for 18-24 h at 35 degrees C and then the colonies were counted . RESULTS: Ropivacaine did not inhibit any of the microorganisms tested . Bupivacaine reduced the viable cells of P . aeruginosa at 0.5% and 0.25% solutions . Lidocaine 5% and 2% and prilocaine 2.0% dilutions reduced the viable cells of all microorganisms tested . Prilocaine 1.0% reduced the viable cells of E . coli, S . aureus and P . aeruginosa . Lidocaine 1% reduced only the viable cells of P . aeruginosa and prilocaine 0.5% reduced only E . coli . CONCLUSION: Ropivacaine had no antimicrobial effect on microorganisms tested . Bupivacaine showed poor antimicrobial effectiveness . Lidocaine and prilocaine had more powerful antimicrobial effects than the other two local anaesthetics.

J Infect, 2001 Apr, 42(3), 208 - 9
Candida albicans prosthetic arthritis treated with fluconazole alone; Merrer J et al.; Conventional treatment of Candida prosthetic joint infection usually includes surgery followed by a long period of antifungal medication . We report a case of Candida albicans prosthetic arthritis successfully treated with fluconazole alone .

J Oral Pathol Med, 2001 Sep, 30(8), 471 - 80
Oral pseudomembranous candidiasis, herpes simplex virus-1 infection, and oral mucositis in head and neck cancer patients receiving radiotherapy and granulocyte-macrophage colony-stimulating factor (GM-CSF) mouthwash; Nicolatou-Galitis O et al.; Oral pseudomembranous candidiasis (OPC) was evaluated in 61 patients receiving head and neck radiotherapy (RT) . Herpes simplex virus-1 (HSV-1) reactivation was also investigated in 14 patients . According to the agreed protocol, granulocyte-macrophage colony-stimulating factor (GM-CSF) mouthwash was administered in 46 patients with radiation-induced ulcers . Candidiasis was diagnosed in 31 patients . Candida albicans was the most frequent isolate . Multiple Candida species were isolated from the lesions of four patients . Concurrent candidiasis and radiation-induced ulcers were observed in 17 patients . Viral culture and the polymerase chain reaction disclosed the presence of HSV-1 in five patients . Twenty of the 46 patients, with initial mucositis grade II and grade III, completed RT with mucositis grade I, indicating a beneficial effect of GMCSF mouthwash, although further controlled studies are necessary to verify that . In conclusion, OPC was an important infection in patients undergoing radiotherapy . The role of HSV-1 in oral mucositis during head and neck radiotherapy needs additional study.

Adv Space Res, 1992, 12(4), 271 - 4
Survival rates of some terrestrial microorganisms under simulated space conditions; Koike J et al.; In connection with planetary quarantine, we have been studying the survival rates of nine species of terrestrial microorganisms (viruses, bacteria, yeasts, fungi, etc.) under simulated interstellar conditions . If common terrestrial microorganisms cannot survive in space even for short periods, we can greatly reduce expenditure for sterilizing space probes . The interstellar environment in the solar system has been simulated by low temperature, high vacuum (77 k, 4 x 10(-6) torr), and protons irradiation from a Van de Graaff generator . After exposure to a barrage of protons corresponding to about 250 years of irradiation in solar space, Tobacco mosaic virus, Bacillus subtilis spores, Aspergillus niger spores and Clostridiun mangenoti spores showed survival rates of 82%, 45%, 28%, and 25%, respectively . Furthermore . pathogenic Candida albicans showed 7% survival after irradiation corresponding to about 60 years in space.

Yeast, 2001 Sep 15, 18(12), 1117 - 29
CaALK8, an alkane assimilating cytochrome P450, confers multidrug resistance when expressed in a hypersensitive strain of Candida albicans; Panwar SL et al.; We report the isolation of a novel C . albicans gene designated CaALK8, by its ability to complement drug hypersensitivity of a pdr5 (ABC: ATP-binding cassette drug extrusion pump) null mutant of S . cerevisiae (JG436) . CaALK8 in JG436 conferred resistance to drugs such as cycloheximide (CYH), fluconazole (FCZ), O-phenanthroline (PHE) and 4-nitroquinoline oxide (NQO) . The gene was so designated because its sequence was identical to a partial sequence entry named as ALK8 in the Candida database . CaALK8 encodes for a putative 515 amino acid protein highly homologous to alkane-inducible cytochromes P450 (CYP52 gene family) of C . maltosa and C . tropicalis . The ability of CaALK8 to confer drug resistance was also established by its expression in another drug-hypersensitive strain of S . cerevisiae (AD 1234568), which was deleted in seven ABC efflux pumps . The homozygous disruption of CaALK8 in a wild-type C . albicans strain (CAI4) did not result in altered drug susceptibilities . The overexpression of CaALK8 in CAI4 resulted in only FCZ resistance . However, a distinct MDR phenotype was evident when CaALK8 was overexpressed in a drug-hypersensitive C . albicans strain disrupted in both CDR1 and CDR2 (ABC drug extrusion pumps of C . albicans) . Alk8p, similar to other Alk proteins from C . maltosa and C . tropicalis, could hydroxylate alkanes and fatty acids . In this study we demonstrate that several drugs could compete with the hydroxylation activity by directly interacting with CaAlk8p . Taken together, our results suggest that a member of the CYP52 gene family could mediate MDR in C . albicans, although it does not seem to be involved in the development of azole resistance in clinical isolates . The nucleotide sequence reported in this paper has been submitted to GenBank under Accession No . Y14766 .

Med Sci Monit, 2001 Sep-Oct, 7(5), 982 - 8
Fungal colonization of gastric mucosa and its clinical relevance; Zwolinska-Wcislo M et al.; BACKGROUND: The aim of our study was to evaluate the incidence of fungi in the stomach in patients with gastric ulcer and chronic gastritis in comparison to healthy humans, and to identify the fungus species isolated from these patients and their susceptibility to antifungal agents . We also assessed the coincidence of the presence of antifungal antibodies and fungal mannan antigen in serum with the concentration of fungi in the stomach . MATERIAL AND METHODS: We investigated 293 patients, aged 20-80, who visited the Gastroenterology Outpatient Clinic at the Jagellonian University's Collegium Medicum in Cracow, complaining of dyspeptic symptoms or clinical manifestations of ulcer disease . The examinations included endoscopy of the upper part of the alimentary tract with sampling of gastric contents, as well as surface brushing and biopsy from the bottom of the ulceration for mycological analysis . Also, biopsy specimens from the margin of the ulceration or inflammatory mucosa were collected for histological examination and urease testing . RESULTS: Gastric mucosa and stomach contents are often an area of fungal colonization, which was detected in 54.2% of the gastric ulcer cases and 10.3% of the chronic gastritis cases . The most frequently isolated fungus species was Candida albicans, although other fungi, previously considered rare or uncommon, were also found . A difference in growth in vitro between the C . albicans, C . tropicalis and C . lusitaniae strains was discovered: C . albicans and C . tropicalis grew from pH 2.0, while C . lusitaniae grew from pH 3.0 . This finding suggests differentiation in the properties of these fungi . CONCLUSIONS: The lack of correlation between the concentration of fungi, the titre of antifungal antibodies and the presence of fungal antigen in serum suggests that fungal colonization is secondary in nature.

Microbiology, 2001 Sep, 147(Pt 9), 2585 - 97
Reintroduction of the PLB1 gene into Candida albicans restores virulence in vivo; Mukherjee PK et al.; Phospholipases have been proposed to contribute to the virulence of Candida albicans . Recently, a candidal strain deleted for PLB1, the gene encoding the predominant phospholipase B (Plb1) secreted by C . albicans, was constructed and its virulence in an intravenous murine model of disseminated candidiasis was evaluated . In the present study, the PLB1 gene was reintroduced back into the plb1 null mutant to generate the revertant strain, which showed similar growth and morphology to its isogenic parent strain . Virulence of the revertant strain was found to be comparable to that of the parent strain in an intravenous murine model of disseminated candidiasis . To compare the abilities of the plb1 null mutant, the revertant and the isogenic parent strains to cross the gastrointestinal (GI) tract and cause systemic infection, an oral-intragastric infant mouse model of candidiasis was used . Histological examinations and analysis of c.f.u . of the pathogen in liver homogenates revealed that the parental and revertant strains were able to invade and traverse the GI mucosa to a significantly greater extent than the plb1 null mutant . Immunofluorescence and immunoelectron microscopic studies of infected host tissue using anti-Plb1 antibody showed that Plb1 is secreted during invasion of the gastric mucosa by the parental and revertant strains . In contrast, little or no labelling was observed in the null mutant strain . The results indicate that the Plb1 secreted by C . albicans enhances the ability of this organism to cross the GI tract and disseminate haematogenously . These studies provide unequivocal evidence supporting a role for Plb1 during the course of infection by C . albicans.

Mol Cell Biol, 2001 Oct, 21(19), 6418 - 28
The basic helix-loop-helix transcription factor Cph2 regulates hyphal development in Candida albicans partly via TEC1; Lane S et al.; Candida albicans undergoes a morphogenetic switch from budding yeast to hyphal growth form in response to a variety of stimuli and growth conditions . Multiple signaling pathways, including a Cph1-mediated mitogen-activated protein kinase pathway and an Efg1-mediated cyclic AMP/protein kinase A pathway, regulate the transition . Here we report the identification of a basic helix-loop-helix transcription factor of the Myc subfamily (Cph2) by its ability to promote pseudohyphal growth in Saccharomyces cerevisiae . Like sterol response element binding protein 1, Cph2 has a Tyr instead of a conserved Arg in the basic DNA binding region . Cph2 regulates hyphal development in C . albicans, as cph2/cph2 mutant strains show medium-specific impairment in hyphal development and in the induction of hypha-specific genes . However, many hypha-specific genes do not have potential Cph2 binding sites in their upstream regions . Interestingly, upstream sequences of all known hypha-specific genes are found to contain potential binding sites for Tec1, a regulator of hyphal development . Northern analysis shows that TEC1 transcription is highest in the medium in which cph2/cph2 displays a defect in hyphal development, and Cph2 is necessary for this transcriptional induction of TEC1 . In vitro gel mobility shift experiments show that Cph2 directly binds to the two sterol regulatory element 1-like elements upstream of TEC1 . Furthermore, the ectopic expression of TEC1 suppresses the defect of cph2/cph2 in hyphal development . Therefore, the function of Cph2 in hyphal transcription is mediated, in part, through Tec1 . We further show that this function of Cph2 is independent of the Cph1- and Efg1-mediated pathways.

EMBO J, 2001 Sep 3, 20(17), 4753 - 61
NRG1, a repressor of filamentous growth in C.albicans, is down-regulated during filament induction; Braun BR et al.; In response to a variety of external signals, the fungal pathogen Candida albicans undergoes a transition between ellipsoidal single cells (blastospores) and filaments composed of elongated cells attached end-to-end . Here we identify a DNA-binding protein, Nrg1, that represses filamentous growth in Candida probably by acting through the co-repressor Tup1 . nrg1 mutant cells are predominantly filamentous under non-filament-inducing conditions and their colony morphology resembles that of tup1 mutants . We also identify two filament-specific genes, ECE1 and HWP1, whose transcription is repressed by Nrg1 under non-inducing conditions . These genes constitute a subset of those under Tup1 control, providing further evidence that Nrg1 acts by recruiting Tup1 to target genes . We show that growth in serum at 37 degrees C, a potent inducer of filamentous growth, causes a reduction of NRG1 mRNA, suggesting that filamentous growth is induced by the down-regulation of NRG1 . Consistent with this idea, expression of NRG1 from a non-regulated promoter partially blocks the induction of filamentous growth.

EMBO J, 2001 Sep 3, 20(17), 4742 - 52
NRG1 represses yeast-hypha morphogenesis and hypha-specific gene expression in Candida albicans; Murad AM et al.; We have characterized CaNrg1 from Candida albicans, the major fungal pathogen in humans . CaNrg1 contains a zinc finger domain that is conserved in transcriptional regulators from fungi to humans . It is most closely related to ScNrg1, which represses transcription in a Tup1-dependent fashion in Saccharomyces cerevisiae . Inactivation of CaNrg1 in C.albicans causes filamentous and invasive growth, derepresses hypha-specific genes, increases sensitivity to some stresses and attenuates virulence . A tup1 mutant displays similar phenotypes . However, unlike tup1 cells, nrg1 cells can form normal hyphae, generate chlamydospores at normal rates and grow at 42 degrees C . Transcript profiling of 2002 C.albicans genes reveals that CaNrg1 represses a subset of CaTup1-regulated genes, which includes known hypha-specific genes and other virulence factors . Most of these genes contain an Nrg1 response element (NRE) in their promoter . CaNrg1 interacts specifically with an NRE in vitro . Also, deletion of two NREs from the ALS8 promoter releases it from Nrg1-mediated repression . Hence, CaNrg1 is a transcriptional repressor that appears to target CaTup1 to a distinct set of virulence-related functions, including yeast-hypha morphogenesis.

J Intraven Nurs, 2001 May-Jun, 24(3), 180 - 205
Guidelines for the management of intravascular catheter-related infections; Mermel LA et al.; These guidelines from the Infectious Diseases Society of America (IDSA), the American College of Critical Care Medicine (for the Society of Critical Care Medicine), and the Society for Healthcare Epidemiology of America contain recommendations for the management of adults and children with, and diagnosis of infections related to, peripheral and nontunneled central venous catheters (CVCs), pulmonary artery catheters, tunneled central catheters, and implantable devices . The guidelines, written for clinicians, contain IDSA evidence-based recommendations for assessment of the quality and strength of the data . Recommendations are presented according to the type of catheter, the infecting organism, and the associated complications . Intravascular catheter-related infections are a major cause of morbidity and mortality in the United States . Coagulase-negative staphylococci, Staphylococcus aureus, aerobic gram-negative bacilli, and Candida albicans most commonly cause catheter-related bloodstream infection . Management of catheter-related infection varies according to the type of catheter involved . After appropriate cultures of blood and catheter samples are done, empirical i.v . antimicrobial therapy should be initiated on the basis of clinical clues, the severity of the patient's acute illness, underlying disease, and the potential pathogen(s) involved . In most cases of nontunneled CVC-related bacteremia and fungemia, the CVC should be removed . For management of bacteremia and fungemia from a tunneled catheter or implantable device, such as a port, the decision to remove the catheter or device should be based on the severity of the patient's illness, documentation that the vascular-access device is infected, assessment of the specific pathogen involved, and presence of complications, such as endocarditis, septic thrombosis, tunnel infection, or metastatic seeding . When a catheter-related infection is documented and a specific pathogen is identified, systemic antimicrobial therapy should be narrowed and consideration given for antibiotic lock therapy, if the CVC or implantable device is not removed . These guidelines address the issues related to the management of catheter-related bacteremia and associated complications . Separate guidelines will address specific issues related to the prevention of catheter-related infections . Performance indicators for the management of catheter-related infection are included at the end of the document . Because the pathogenesis of catheter-related infections is complicated, the virulence of the pathogens is variable, and the host factors have not been well defined, there is a notable absence of compelling clinical data to make firm recommendations for an individual patient . Therefore, the recommendations in these guidelines are intended to support, and not replace, good clinical judgment . Also, a section on selected, unresolved clinical issues that require further study and research has been included . There is an urgent need for large, well-designed clinical studies to delineate management strategies more effectively, which will improve clinical outcomes and save precious health care resources.

Clin Infect Dis, 2001 Oct 1, 33(7), 1069 - 75 Epub 2001 Sep 05.
Evolution of antifungal susceptibility among Candida species isolates recovered from human immunodeficiency virus-infected women receiving fluconazole prophylaxis; Vazquez JA et al.; The effect of fluconazole on the susceptibility of Candida isolates recovered from women infected with human immunodeficiency virus (HIV) was evaluated in a randomized, double-blind, placebo-controlled trial . Women with CD4(+) cell counts of < or =300 cells/mm(3) received either fluconazole (200 mg/week) or placebo as prophylaxis . The antifungal susceptibility of specimens was evaluated . One patient who received fluconazole and 2 patients assigned to placebo had Candida albicans isolates recovered that were resistant to fluconazole (MIC, > or =64 microg/mL) . Eleven patients assigned fluconazole and 4 patients assigned placebo had non-albicans Candida strains (all Candida glabrata) recovered that were resistant to fluconazole . There was significant azole cross-resistance among the non-albicans Candida species isolates . Although the rate of azole resistance did not significantly increase after fluconazole prophylaxis, there was a trend toward more in vitro azole resistance in C . glabrata isolates from patients assigned fluconazole . Moreover, the majority of resistant vaginal isolates of Candida species were recovered after initiation of open-label fluconazole use.

Clin Diagn Lab Immunol, 2001 Sep, 8(5), 943 - 8
Highly active antiretroviral therapy in human immunodeficiency virus type 1-infected children: analysis of cellular immune responses; Blazevic V et al.; The present study analyzes the effect of highly active antiretroviral therapy (HAART) on restoration of cellular immunity in human immunodeficiency virus (HIV)-infected children over a 24-week period following initiation of HAART with ritonavir, nevirapine, and stavudine . The immunological parameters evaluated at four time points (at enrollment and at 4, 12, and 24 weeks of therapy) included cytokine production by monocytes as well as T-cell proliferation in response to mitogen, alloantigen, and recall antigens including HIV type 1 envelope peptides . Circulating levels of interleukin-16 (IL-16) were measured, in addition to CD4+ T-cell counts, plasma HIV RNA levels, and the delayed-type hypersensitivity (DTH) response . At enrollment the children exhibited defects in several immune parameters measured . Therapy increased CD4+ T-cell counts and decreased viral loads significantly . By contrast, the only immunological parameter that was significantly increased was IL-12 p70 production by monocytes; the DTH response to Candida albicans also showed a strong increase in patients becoming positive . In conclusion, these results demonstrate that HAART in HIV-infected children affects the dynamics of HIV replication and the CD4+ T-cell count over 24 weeks, similar to the pattern seen in HIV-infected adults . Furthermore, these data indicate improvement in antigen-presenting cell immunological function in HIV-infected children induced by HAART.

J Clin Microbiol, 2001 Sep, 39(9), 3362 - 4
Discrimination between Candida albicans and other pathogenic species of the genus Candida by their differential sensitivities to toxins of a panel of killer yeasts; Buzzini P et al.; The differential sensitivities to toxins produced by a short panel of four killer yeasts allowed discrimination between 91 strains of the yeast Candida albicans and 223 non-C . albicans Candida strains . One hundred percent of C . albicans isolates exhibited negative results to the toxin panel, while 100% of non-C . albicans cultures gave well-defined and reproducible positive results to at least one of the four killer toxins . Among C . albicans strains only 96 and 87% gave germ tube (GT)- and chlamydospore-positive results, respectively . In addition a few GT-false-positive strains were detected among non-C . albicans isolates . Susceptibility to the toxin panel is apparently expressed more consistently than either GT or chlamydospore production and may constitute a promising basis for a new simple and easy-to-use procedure for routine discrimination between the species C . albicans and other species of the genus Candida.

J Clin Microbiol, 2001 Sep, 39(9), 3296 - 302
Antifungal effects of lysozyme and lactoferrin against genetically similar, sequential Candida albicans isolates from a human immunodeficiency virus-infected southern Chinese cohort; Samaranayake YH et al.; A variety of innate defense factors in saliva such as lysozyme and lactoferrin contribute to mucosal protection and modulate Candida populations in the oral cavity . It is also known that in human immunodeficiency virus (HIV)-infected individuals significant variations in the concentrations of lysozyme and lactoferrin in saliva occur during disease progression . Therefore, the aim of this study was to determine the in vitro susceptibility to human lactoferrin and hen egg white lysozyme of genotypically similar oral Candida albicans isolates obtained from six HIV-infected ethnic Chinese during sequential visits over a 12-month period . The similarity of the genotypes (50 in total) was evaluated using a randomly amplified polymorphic DNA assay . A blastospore viability assay was performed to evaluate the sensitivity of the organisms to lysozyme and lactoferrin . Exposure to physiological concentrations of either lysozyme (30 microg/ml) or lactoferrin (20 microg/ml) caused a rapid loss of viability among all isolates to a varying extent . None of the sequential C . albicans isolates demonstrated significant differences in sensitivity to either protein from one visit to the next; similar results were noted when the different genotypes from the same individual were compared . On Spearman correlation analysis of two genotypes that were sequentially isolated from a single patient, a significant negative correlation between lysozyme (r = -0.88; P < 0.02) (but not lactoferrin) resistance and the duration of HIV disease was seen . These results imply that a minority of C . albicans isolates that persist intraorally in individuals with HIV disease develop progressive resistance to innate salivary antifungal defenses such as lysozyme, possibly as an adaptive response . However, the vast majority of the Candida isolates appear to succumb to these nonspecific host immune mediators abundantly present in the oral environment.

Appl Environ Microbiol, 2001 Sep, 67(9), 4030 - 5
Toxic effects of Ag(I) and Hg(II) on Candida albicans and C . maltosa: a flow cytometric evaluation; Zhang S et al.; The effects of Ag(I) and Hg(II) on membrane potential and integrity of cells of Candida albicans and C . maltosa were determined with a flow cytometric procedure that employed an anionic membrane potential-sensitive dye, bis-(1,3-dibutylbarbituric acid) trimethine oxonol, and a membrane integrity indicator, propidium iodide . The membrane potentials of cells of both species were reduced rapidly within 15 min of exposure to Ag(I) . No threshold dose for Hg(II) existed, and cells of both species lost membrane potential gradually in Hg(II) solutions . Cells of both species lost membrane integrity more rapidly in Ag(I) solutions than in Hg(II) solutions . In Ag(I) solutions, the decrease in the numbers of cells recoverable in culture occurred at a rate similar to the rate of cell depolarization and membrane permeabilization . In Hg(II) solutions, loss of cell recoverability preceded the loss of membrane potential and membrane integrity . C . albicans, in contrast to C . maltosa, showed no loss of membrane integrity after exposure to Hg(II) solutions for 1 h . Different rates of binding of Ag(I) and Hg(II) between the two species suggest that the two ions target different primary sites.

Mutat Res, 2001 Oct 18, 497(1-2), 213 - 22
Antioxidative and antimutagenic activity of yeast cell wall mannans in vitro; Krizkova L et al.; Antioxidative and antimutagenic effect of yeast cell wall mannans, in particular, extracellular glucomannan (EC-GM) and glucomannan (GM-C.u.) both from Candida utilis, mannan from Saccharomyces cerevisiae (M-S.c.) and mannan from Candida albicans (M-C.a.) was evaluated . Luminol-dependent photochemical method using trolox as a standard showed that EC-GM, GM-C.u., M-S.c . and M-C.a . have relatively good antioxidative properties . EC-GM exhibited the highest antioxidative activity, followed by GM-C.u . and M-S.c . M-C.a . showed the least antioxidative activity . These mannans were experimentally confirmed to exhibit different, statistically significant antimutagenic activity in reducing damage of chloroplast DNA of the flagellate Euglena gracilis induced by ofloxacin and acridine orange (AO) . We suggest that the antimutagenic effect of EC-GM, GM-C.u., M-S.c . and M-C.a . against ofloxacin is based on their ability to scavenge reactive oxygen radicals . With AO, the reduction of the chloroplast DNA lession could be a result of the absorptive capacity of the mannans . The important characteristics of mannans isolated from the yeast cell walls, such as good water solubility, relatively small molecular weight (15-30kDa), and antimutagenic effect exerted through different mode of action, appear to be a promising features for their prospective use as a natural protective (antimutagenic) agents.

J Asian Nat Prod Res, 1999, 1(4), 269 - 75
A triterpene from Ficus pumila; Ragasa CY et al.; The leaves of Ficus pumila afforded a new neohopane (1) by silica gel chromatography . The structure of 1 was elucidated by 1D and 2D NMR and IR spectroscopy and mass spectrometry . It showed antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Candida albicans with an average antimicrobial index of 0.5, 0.3, 0.3 and 0.7, respectively, at a concentration of 30 microg.

Eur J Pharm Biopharm, 2001 Sep, 52(2), 255 - 9
A case study of preservation of semi-solid preparations using the European Pharmacopoeia test: comparative efficacy of antimicrobial agents in zinc gelatin; Favet J et al.; The present study was undertaken with the aim of finding an alternative preservative system to methyl parahydroxybenzoate in zinc gelatin, which was described in the monographs of the Swiss Pharmacopoeia (until Ph . Helv . 8) and in previous editions of the German Pharmacopoeia (until DAB 7) . This antimicrobial agent has now been withdrawn in the DAB, because of its potential allergy risks . As for the USP and DAB-DDR zinc gelatin preparations, they have always been devoid of any preservative agent, probably relying on the mild antimicrobial activity of zinc . A literature survey did not reveal if such an aqueous preparation containing the water-insoluble zinc oxide shows efficacious antimicrobial activity by itself . Thus, a comparative evaluation of differently preserved zinc gelatin preparations was performed using a test for the efficacy of antimicrobial preservation that has been modified with regard to the European Pharmacopoeia (EP) test to take into account the solid state of the preparations and the bactericidal effect of the zinc . Three zinc gelatin preparations were checked, either: (i), without any agent; or (ii), with 0.1% methyl parahydroxybenzoate; or (iii), with 0.5% phenoxyethanol, a broad-spectrum antimicrobial agent almost devoid of allergy risks . The three preparations behave quite differently, in particular with respect to fungi . All three preparations passed the modified EP test as far as bacteria are concerned . Even zinc gelatin without preservative is very effective, not only because of the mild antimicrobial activity of zinc (the soluble fraction of zinc oxide in the liquid phase of zinc gelatin was determined to be 13 microg/ml), but most probably because of the low water activity of the preparation (measured as around 0.81), as shown by the absence of growth of a zinc-resistant strain of Pseudomonas aeruginosa . Zinc gelatin preserved with methyl parahydroxybenzoate has a weak, although satisfactory, activity against Staphylococcus aureus . Regarding fungi, gelatin without an antimicrobial agent and that preserved with methyl parahydroxybenzoate meet the requirements for efficacy against Candida albicans, but are only bacteriostatic against Aspergillus niger . As for zinc gelatin preserved with phenoxyethanol, it displays the best activity against C . albicans and, above all, appears to be the only formulation exhibiting fungicidal activity against A . niger . It is therefore recommended to preserve zinc gelatin with this antimicrobial agent, as recently adopted in Supplement 2000 of the Swiss Pharmacopoeia.

FEBS Lett, 2001 Aug 24, 504(1-2), 11 - 5
The Candida albicans Na(+)/H(+) antiporter exports potassium and rubidium; Kinclova O et al.; The Candida albicans Cnh1p belongs to the family of Na(+)/H(+) antiporters (TC 2.A.36) but it transports besides toxic sodium and lithium also rubidium and potassium . Upon heterologous expression in a Saccharomyces cerevisiae salt-sensitive strain, the Cnh1p is targeted to the plasma membrane and its transport activity results in increased tolerance of cells to external alkali metal cations . The cation efflux activity of Cnh1p in S . cerevisiae depends on the gradient of protons across the plasma membrane, and a Cnh1p-mediated K(+) efflux is involved in a cell response to sudden rise of cytoplasmic pH.

J Med Chem, 2001 Aug 30, 44(18), 2928 - 32
X-Ray crystal structures of Candida albicans dihydrofolate reductase: high resolution ternary complexes in which the dihydronicotinamide moiety of NADPH is displaced by an inhibitor; Whitlow M et al.; X-ray crystallographic analysis of 5-(4'-substituted phenyl)sulfanyl-2,4-diaminoquinazoline inhibitors in ternary complex with Candida albicans dihydrofolate reductase (DHFR) and NADPH revealed two distinct modes of binding . The two compounds with small 4'-substituents (H and CH3) were found to bind with the phenyl group oriented in the plane of the quinazoline ring system and positioned adjacent to the C-helix . In contrast, the more selective inhibitors with larger 4'-substituents (tert-butyl and N-morpholino) were bound to the enzyme with the phenyl group perpendicular to the quinazoline ring and positioned in the region of the active site that typically binds the dihydronicotinamide moiety of NADPH . The cofactor appeared bound to DHFR but with the disordered dihydronicotinamide swung away from the protein surface and into solution . This unusual inhibitor binding mode may play an important role in the high DHFR selectivity of these compounds and also may provide new ideas for inhibitor design.

Am J Obstet Gynecol, 2001 Aug, 185(2), 363 - 9
Treatment of complicated Candida vaginitis: comparison of single and sequential doses of fluconazole; Sobel JD et al.; OBJECTIVE: An attempt was made to validate recent recommendations that women with complicated Candida vaginitis (severe or recurrent, non-albicans Candida spp or abnormal host) require longer-duration antifungal therapy to achieve clinical cure and mycologic eradication . STUDY DESIGN: A prospective, multicenter, randomized, double-blind study was performed comparing a single dose of 150 mg of fluconazole with 2 sequential 150-mg doses of fluconazole given 3 days apart . RESULTS: Five hundred fifty-six women with severe or recurrent Candida vaginitis were enrolled, and 398 had at least one postbaseline evaluation (intent to treat) and of these 309 were fully evaluable (efficacy-valid) . At baseline, 92% of vaginal isolates were Candida albicans . The 2-dose fluconazole regimen achieved significantly higher clinical cure rates in women with severe vaginitis when evaluated on day 14 (P =.015) and higher clinical and mycologic responses persisted at day 35 . Women with recurrent but not severe vaginitis did not benefit clinically short term by the additional fluconazole dose . Multivariate logistic regression analysis showed that being infected with non-albicans Candida predicted significantly reduced clinical and mycologic response regardless of duration of therapy . Fluconazole therapy was well tolerated and free of serious adverse effects . CONCLUSION: Treatment of Candida vaginitis requires individualization, and women with severe Candida vaginitis achieve superior clinical and mycologic eradication with a 2-dose fluconazole regimen.

J Hosp Infect, 2001 Sep, 49(1), 37 - 42
Rate of transmission and endogenous origin of Candida albicans and Candida glabrata on adult intensive care units studied by pulsed field gel electrophoresis; Hamal P et al.; We determined the relative roles of endogenous origin and patient-to-patient transmission in Candida colonization of patients on adult intensive care units (ICU) . A total of 48 Candida albicans and 18 Candida glabrata strains from various clinical samples of 28 long-term patients, hospitalized in two neurological ICUs between April and June 1999, were typed using pulsed field gel electrophoresis (PFGE) . Three patients were co-colonized by both C . albicans and C . glabrata strains . Twenty-four C . albicans and 17 C . glabrata karyotypes were defined . The colonization was found to be polyclonal in six C . albicans and five C . glabrata patients . Twenty-six patients (93%) carried strains, which were not detected in other patients hospitalized at the same time, i.e . they were colonized by unique C . albicans and C . glabrata strains . Only two patients, who were hospitalized during the same period of time, although in different rooms of the same ICU, shared strains with an identical PFGE type, indicating possible patient-to-patient transmission . Patient-to-patient transmission of yeasts played a minor role on these ICUs .

J Invest Dermatol, 2001 Aug, 117(2), 205 - 13
A mannose-binding receptor is expressed on human keratinocytes and mediates killing of Candida albicans; Szolnoky G et al.; Human keratinocytes are known to kill Candida albicans in vitro, but the mechanism of killing is not yet understood . Here, we demonstrate that spontaneous, ultraviolet-B-light-induced, alpha-melanocyte-stimulating-hormone-induced, and interleukin-8-induced Candida killing by keratinocytes can be inhibited with mannan and mannosylated bovine serum albumin (Man-BSA) . A polyclonal goat serum raised against the human macrophage mannose receptor stained suprabasal keratinocytes, but no staining was observed on keratinocytes with a monoclonal antibody (mAb15) specific for the human macrophage mannose receptor . Mannose-affinity chromatography of keratinocyte extract isolated a 200 kDa protein, and on the Western blot the goat antiserum reacted with a 200 kDa protein . In radioligand binding studies, the binding of 125I-Man-BSA to human keratinocytes was inhibited by mannan in a concentration-dependent manner . Analysis of the binding revealed a single class keratinocyte mannose receptor with a KD of 1.4 x 10(-8) M and a Bmax of 1 x 10(4) binding sites per cell . The binding of 125I-Man- BSA to keratinocytes proved to be time-dependent, acid-precipitable, and Ca2+- and trypsin-sensitive . After trypsinization the receptors underwent a rapid recovery at 37 degrees C . These results demonstrate the presence of mannose receptor on human keratinocytes, and its active involvement in the killing of Candida albicans.

Biomaterials, 2001 Sep, 22(17), 2319 - 24
A polymeric system for the intra-oral delivery of an anti-fungal agent; Patel MP et al.; Oral candidal infections are often persistent and intractable and thus the aim of this study was to develop a polymeric sustained release device to improve the topical treatment of these infections . A self curing system based on poly(ethyl methacrylate) and tetrahydrofurfuryl methacrylate (PEM/THFM) was used with chlorhexidine diacetate (CX) added at levels between 0 and 12% w/w . Water uptake by the device was assessed gravimetrically and CX release measured by UV spectrometry . Anti candidal activity was established by culturing azole sensitive and resistant strains of Candida albicans in the presence of the polymeric delivery device with and without CX . Candidal growth was measured by turbidimetry or surviving colony-forming unit (CFU) formation . There was an initial high release of CX over 24 h followed by a slow diffusion up to 7 days . CX inhibited candidal growth and survival markedly in vitro, with the test samples showing less than 0.5 x 10(-7) CFU/ml compared to controls (3-4 x 10(-7) CFU/ml) . These results indicate the potential of a chlorhexidine containing PEM/THFM polymeric system in the treatment of persistent candidal infections.

Adv Perit Dial, 2001, 17, 176 - 9
Non Candida albicans fungal peritonitis in continuous ambulatory peritoneal dialysis patients; Kleinpeter MA et al.; We report four episodes of non Candida albicans peritonitis (NCAP) in 3 patients on continuous ambulatory peritoneal dialysis (CAPD) . Risk factors for NCAP included diabetes mellitus and prior antibiotic use in half of the cases . The antibiotic treatment was prescribed for exit-site infection (ESI) or peritonitis in the patient . Treatment for NCAP included antifungal therapy with oral fluconazole or intravenous amphotericin B . The NCAP resulted in catheter loss in 100% of the patients over time . Initial catheter salvage in one patient was followed 6 months later by catheter loss following treatment of a bacterial peritonitis that was complicated by the development of Candida (Torulopsis) glabrata peritonitis unresponsive to treatment with intravenous amphotericin B . Although the literature suggests that Candida peritonitis responds to oral fluconazole with and without catheter removal, this series suggests that the treatment of NCAP includes removal of the peritoneal dialysis catheter with appropriate antifungal agents.

Phytother Res, 2001 Aug, 15(5), 401 - 6
Screening of Malian medicinal plants for antifungal, larvicidal, molluscicidal, antioxidant and radical scavenging activities; Diallo D et al.; A total of 78 different extracts from 20 medicinal plants belonging to 14 plant families from Mali were tested for their antifungal, larvicidal, molluscicidal, antioxidant and radical scavenging activities . Dichloromethane, methanol, water and ethanol extracts were used . TLC autobiography for antifungal activity was run with Cladosporium cucumerinum and Candida albicans . Extracts were also tested on the larvae of the mosquitoes Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus . Molluscicidal activities were established with the snails Biomphalaria glabrata, Biomphalaria pfeifferi and Bulinus truncatus . beta-Carotene and DPPH solutions sprayed on TLC plates were used for antioxidant and radical scavenging assays . Of the extracts investigated, 20% were antioxidant and radical scavengers, 19% fungicidal, 30% were larvicidal and 11% were molluscicidal . Three of the plant extracts, from Cussonia barteri (Araliaceae), Glinus oppositifolius (Aizoaceae) and Lannea velutina (Anacardiaceae) gave positive responses in all four tests .

Clin Chem Lab Med, 2001 Jun, 39(6), 519 - 26
Increased serum and salivary immunoglobulins against Candida albicans in HIV-infected patients with oral candidiasis; Drobacheff C et al.; The aim of this study was to explore anti-Candida albicans systemic and mucosal humoral responses against Candida virulence antigens such as somatic antigen and secreted aspartic proteases (Saps) in HIV-infected patients with oral candidiasis . Twenty-eight subjects were included in the study: 11 HIV-positive patients without oral candidiasis (group A), 6 HIV-positive patients with oral candidiasis (group B) and 11 HIV-negative healthy controls (group C) . Total IgA, IgG and IgM concentrations and antibodies to C . albicans (somatic antigen, Sap1, Sap6) were measured in serum and saliva . We developed a time-resolved immunofluorometric assay with biotin and europium-labeled streptavidin for this purpose . Salivary total IgA, IgG and IgM concentrations were higher in group B . IgA, IgG and IgM anti-C . albicans antibodies (against somatic antigen, Sap1, Sap6) were higher in saliva and serum from patients from group B compared with patients from group A and controls . Our results suggest that, in oral candidiasis, HIV-infected patients have a high mucosal response, specifically directed against C . albicans virulence antigens, such as somatic antigen, Sap1 and Sap6.

Drug Resist Updat, 1999 Feb, 2(1), 9 - 14
Antifungal resistance in non- albicans Candida species; Collin B et al.; Non- Candida albicans species have emerged as important bloodstream pathogens . They tend to have decreased susceptibility to antifungal agents in vitro and cause infections associated with high morbidity and mortality . Fluconazole resistance can emerge in any Candida spp., but is most commonly seen with Candida krusei, for which resistance is universal, and with Candida glabrata . Amphotericin B resistance has also been increasingly reported, most notably in isolates of Candida lusitaniae and Candida guilliermondii . Efforts are underway to correlate in-vitro antifungal susceptibility of individual Candida isolates with response to therapy of patients with candidemia . Future advances in this field might allow physicians to identify Candida isolates resistant to specific antifungal agents and thereby tailor therapy of candidemia .

Transplantation, 2001 Aug 15, 72(3), 477 - 9
Candida fasciitis following renal transplantation; Wai PH et al.; BACKGROUND: We describe a rare case of necrotizing fasciitis involving Candida albicans, an organism that has been reported to have a minimal potential for invasive soft tissue infection . In this case, immunosuppression, chronic renal failure, and a history of diabetes mellitus were predisposing factors . METHODS: The medical record and histopathologic material were examined . The clinical literature was reviewed for previous cases of C albicans necrotizing fasciitis . RESULTS: A review of the literature showed that in solid organ transplant recipients, localized fungal soft tissue infection is infrequent, with only 35 cases reported between 1974 and 1992 . Necrotizing fasciitis caused by C albicans is extremely rare in the modern era of solid organ transplantation . CONCLUSIONS: The management of transplant patients at risk for invasive fungal infection warrants a high index of suspicion for fungal necrotizing fasciitis in the setting of wound infection and merits a thorough investigation for atypical pathogens.

Diagn Microbiol Infect Dis, 2001 Jul, 40(3), 121 - 3
Candida albicans spinal epidural abscess secondary to prosthetic valve endocarditis; Liang JD et al.; A 56-year-old woman, with underlying rheumatic heart disease status post mitral valve replacement, presented with fever, low back pain radiating to right leg, and congestive heart failure . Magnetic resonance imaging detected an L5-S1 spinal epidural abscess . A vegetation on prosthetic mitral valve was found by transesophageal echocardiography . Cultures of epidural aspirate, surgical specimen, and blood all grew Candida albicans . She received surgical drainage of the spinal epidural abscess and i.v . amphotericin B 1 mg/kg/day for eight weeks . Clinical symptoms improved gradually and she was discharged without neurologic sequelae . She remained well and continued to lead an active life two years after discharge.

Eur J Biochem, 2001 Aug, 268(16), 4449 - 58
Internalization of tenecin 3 by a fungal cellular process is essential for its fungicidal effect on Candida albicans; Kim DH et al.; Tenecin 3 is a glycine-rich, antifungal protein of 78 residues isolated from the insect Tenebrio molitor larva . As an initial step towards understanding the antifungal mechanism of tenecin 3, we examined how this protein interacts with the pathogenic fungus Candida albicans to exert its antifungal action . Tenecin 3 did not induce the release of a fluorescent dye trapped in the artificial membrane vesicles and it did not perturb the membrane potential of C . albicans by the initial interaction . Fluorescence confocal microscopy and flow cytometric analysis revealed that tenecin 3 is rapidly internalized into the cytoplasmic space in energy-dependent and temperature-dependent manners . This internalization is also dependent on the ionic environment and cellular metabolic states . These results suggest that the internalization of tenecin 3 into the cytoplasm of C . albicans is mediated by a fungal cellular process . The internalized tenecin 3 is dispersed in the cytoplasm, and the loss of cell viability occurs after this internalization.

Mycopathologia, 2001, 151(1), 5 - 10
Susceptibility profile of vaginal yeast isolates from Brazil; Ribeiro MA et al.; Vaginal specimens for culture were obtained from two hundred and five immunocompetent, non-hospitalized patients selected among all women attending the Gynecology and Obstetric Ambulatory Clinic of the University of Espirito Santo, Brazil, during a 2-year period (From 1998 to 1999) . Patients were checked for signs and symptoms of vulvovaginitis and previous use of topical and systemic antifungal drugs . Yeast isolates were identified by classical methods and the antifungal susceptibility profile was determined according to NCCLS microbroth assay . The prevalence of vaginal yeast isolates from asymptomatic women was 25% (30/121) and 60% (50/84) among patients with symptoms of vulvovaginitis . Candida albicans was the most frequently isolated species in both groups (46% and 90%, respectively), followed by C . glabrata (13% and 6%, respectively) . All isolates were susceptible to amphotericin B . Only ten isolates had dose dependent susceptibility (DDS) or resistance to azoles; and seven of these were non-albicans species . Based on our results we suggest that species identification and antifungal susceptibility testing need not be routinely performed in immunocompetent women, and may be reasonable only for the minority of patients with complicated vulvovaginal candidiasis that fail to respond to therapy.

Zhonghua Zheng Xing Shao Shang Wai Ke Za Zhi, 1999 Mar, 15(2), 105 - 8
{Effect of different immunomodulators on macrophage function in rats with scald injury}; Zhuo A et al.; OBJECTIVE: To explore effect of different immunomodulators on macrophage (M phi) functions in rats with scald injury . METHODS: The M phi functions in various groups of rats with scald on the 5th and 10th day after scald were respectively determined with methods of McAb APAAP, agar bacteriolytic plate and MTT colorimetry . RESULTS: The results showed that: 1 . after scald, presenting rate of Ia antigen, ability of antigen presentation, phagocytic power to candida albicans and lysozyme ability of M phi were significantly decreased, the capacity of M phi secreting TNF was markedly increased compared with those of the normal group, and the differences were significant (P < 0.01); 2 . after the therapy with immunodulators, the above functions of M phi in rats with scald improved markedly . As compared with those of the control group, differences were significant (P < 0.01) . CONCLUSION: The specific immune RNA, which is administered by intraperitoneal injection early after scald, can markedly improve immune functions of rats with scald.

Drug Resist Updat, 1999 Dec, 2(6), 403 - 414
Regulation of pleiotropic drug resistance in yeast; Kolaczkowska A et al.; This review focuses on the molecular mechanisms involved in the regulation of multiple drug resistance in the model yeast Saccharomyces cerevisiae and the pathogenic fungus Candida albicans . Recent developments in the study of the transcription factors Pdr1p, Pdr3p and Yap1p are reported . Understanding the molecular basis leading to multiple drug resistance is a prerequisite for the development of new antifungal therapeutics .

J Clin Pediatr Dent, 2001 Summer, 25(4), 333 - 6
Hyper-IgE syndrome: a case report; Sepet E et al.; The hyper-IgE syndrome (HIES) is a rare disorder characterized by pruritic dermatitis, recurrent Staphylococcus skin abscesses and extremely elevated levels of IgE in serum . In this report, an eleven-year-old-boy with hyper-IgE syndrome is presented . He had a coarse facial appearance, pruritic dermatitis, recurrent skin abscesses, pulmonary infection, a reduced rate of resorption of the roots of primary teeth and an elevated serum IgE concentration . The colonization of Candida albicans, Kiebsiella pneumoniae, Escherichia coli and Staphylococcus aureus were found as; (1x10(2) CFU), (2.2x10(4) CFU), (2.2x10(4) CFU) and (2.6x10(3) CFU) per ml saliva, respectively . Also the pulp of a deciduous molar was investigated with light and transmission electron microscope (TEM) . As conclusion, treatment for this condition is lifelong administration of therapeutic doses of a penicillinase-resistant penicillin, with the addition of other antibiotics or anti-fungal agents as required for specific infections.

Microbiology, 2001 Aug, 147(Pt 8), 2007 - 19
A GAS-like gene family in the pathogenic fungus Candida glabrata; Weig M et al.; In fungi, the cell wall plays a major role in host-pathogen interactions . Despite this, little is known about the molecular basis of cell wall assembly in Candida glabrata, which has emerged as the second most common cause of systemic candidosis . A C . glabrata gene family, CgGAS1-3, that shares significant homologies with both the GAS1 gene of Saccharomyces cerevisiae, which is necessary for cell wall assembly, and the pH-regulated genes PHR1 and PHR2 of Candida albicans, which are involved in cell wall assembly and required for virulence, has been cloned . Among the members of this family, CgGAS1-3 display a unique expression pattern . Both CgGAS1 and CgGAS2 are constitutively expressed . In contrast, CgGAS3 transcript was not detectable under any of the assayed conditions . The C . glabrata actin gene, CgACT1, has also been cloned to be used as a meaningful loading control in Northern blots . CgGAS1 and CgGAS2 were deleted by two different methodological approaches . A rapid PCR-based strategy by which gene disruption was achieved with short regions of homology (50 bp) was applied successfully to C . glabrata . DeltaCggas1 or DeltaCggas2 cells demonstrated similar aberrant morphologies, displaying an altered bud morphology and forming floccose aggregates . These phenotypes suggest a role for CgGAS1 and CgGAS2 in cell wall biosynthesis . Further evidence for this hypothesis was obtained by successful functional complementation of a gas1 null mutation in S . cerevisiae with the C . glabrata CgGAS1 or CgGAS2 gene.

Infect Dis Obstet Gynecol, 2001, 9(2), 65 - 73
Candidiasis during pregnancy may result from isogenic commensal strains; Daniels W et al.; OBJECTIVE: Our laboratory previously demonstrated that asymptomatic vaginal colonization during pregnancy is a factor predisposing patients to subsequent symptomatic vulvovaginal candidiasis . It is unknown whether symptoms result from strain replacement or a change in host relationship to the original colonizing strain . This study was undertaken to determine whether Candida albicans isolates from asymptomatic women could be responsible for subsequent symptomatic vaginitis . METHODS: We retained isolates of C . albicans from women followed longitudinally through pregnancy, and identified six pairs of cultures from women who were colonized without symptoms and who later became symptomatic (average time 14 weeks) . We used a random amplification of polymorphic DNA (RAPD) analysis to determine whether isolates from our study patients were genetically similar or dissimilar . RESULTS: Analysis of these pairs of yeast strains by RAPD revealed that five of the six women had symptoms apparently due to the same yeast strain that was found initially as a commensal strain . To increase the power of these observations, we also performed RAPD analysis on six randomly selected yeast strains from other women in this study who had not become symptomatic to determine whether any of these unrelated strains matched strains from those women who became symptomatic . CONCLUSION: Symptomatic yeast vaginitis is usually due to strains of C . albicans already carried in the lower genital tract, underscoring the need to understand regulation of growth and virulence of the organism in vivo.

Urol Int, 2001, 67(2), 186 - 8
Prostatic abscess due to Candida with no systemic manifestations; Collado A et al.; Prostatic abscess due to fungi is a rare condition . It is generally secondary to systemic disease in immunosuppressed patients . It usually occurs with affection of other organs in a septic patient . Only in exceptional cases does it occur isolatedly . We present the case of a prostatic abscess due to Candida albicans with no systemic manifestations . The diagnosis is helped by transrectal ultrasound, which allows to differentiate this condition from nonabscessed acute prostatitis . The treatment of choice is ultrasound-guided transrectal needle aspiration after antibiotic therapy has been started . As with abscesses of bacterial origin, an ultrasonographic follow-up is required due to the possibility of persistence or recurrence .

J Pharm Biomed Anal, 2001 Oct, 26(3), 387 - 99
Formulation and evaluation of itraconazole via liquid crystal for topical delivery system; Nesseem DI; Liquid crystal systems are used to tailor drug delivery from topical delivery system . Ternary polyoxyethylene {21} stearyl ether/ oil and water form liquid crystalline system cream, which has a potential as dosage form for 1% itraconazole as topical dermal drug delivery . Evaluation of the suggested formula was performed for the best physical performance, the compatibility of the components of lyotropic liquid crystal with itraconazole was conducted through polarized light microscopy, differential scanning calorimeter, thermogravimetric analysis and viscosity measurements . Fourier transform infrared spectroscopy has been studied . Furthermore, in vitro antimycotic inhibitory activity of 1% itraconazole from liquid crystal, was conducted using agar-cup method and Candida albicans as a test organism . The pH value of the cream was found to be 7.1, while when the drug was incorporated in the cream, the pH value was 6.7 . The formula was examined under polarized microscope at 20x magnification and the birefringence that is characteristic of concentric lamellar liquid crystal was observed around the oil globules . Differential scanning calorimeter of itraconazole cream showed higher transition peak temperature at 120 degrees C for the hydrophilic gel phases . Fourier transform infrared spectroscopy revealed that there was no complex or any interaction between the surfactant and the drug . The microbial studies revealed that our formula had the highest zone of inhibition . The average +/- SD inhibition zone values of the test, control I and control II are 30.4+/-1.14, 19.6+/-1.14 and 14.8+/-0.83 mm, respectively . It was found that the test was significantly different from control I and control II, P=5.33x10(-6), 8.92x10(-5), respectively, so it may be concluded that incorporation of the drug in liquid crystal increased its antimicotic activity against Candida albicans.

Planta Med, 2001 Jul, 67(5), 428 - 31
Acidic polysaccharides from rhizomes of Atractylodes lancea as protective principle in Candida-lnfected mice; Inagaki N et al.; Prophylactic effects upon imunnosuppressed mice lethally infected by Candida albicans were examined in fractions prepared from a constituent herb of Juzen-taiho-to (TJ-48, Si-Quan-Da-Bu-Tang), rhizomes of Atractylodes lancea DC . The oral administration of water extract obtained from a residue after MeOH extraction of rhizomes significantly prolonged the survival period of the infected mice at a dose of 140 mg/kg/day compared with control mice, while the MeOH extract did not . In the crude polysaccharide fraction (F-2) obtained by EtOH precipitation of the water extract, a significant life-prolonging effect was observed by the administration of 70 mg/kg/day . F-2 was further fractionated, and the resulting strongly acidic polysaccharide fraction, F-2-2, had a protective effect at a dose of 17.5 mg/kg/day . This fraction mainly consisted of acidic pectic polysaccharides containing about 80% galacturonic acid . The protective activity of F-2-2 was lost by periodate oxidation, but not by protease digestion, suggesting that the polysaccharide component of F-2-2 plays a major role in the protective activity against Candida-infected mice.

J Endod, 2001 Jun, 27(6), 401 - 3
Effect of sodium hypochlorite and five intracanal medications on Candida albicans in root canals; Valera MC et al.; The aim of this study was to evaluate the effect of 1% sodium hypochlorite and five intracanals medications on Candida albicans harvested inside root canals . The contaminated canals were irrigated with sterile saline solution and then treated as follows: (i) filled with Calen paste (calcium hydroxide/ glycol polyethylene paste); (ii) filled with camphorated paramonochloro phenol (CPMC); (iii) filled with 2% iodine-iodate solution; (iv) filled with tricresol formalin; (v) filled with Calen and CPMC pastes; (vi) irrigation with 1% sodium hypochlorite and filled with no intracanal medication; and (vii) no intracanal medication was used . Canal access and the apical foramen were then sealed with Cavit and the roots were stored in a humid chamber at 37 +/- 1 degree C for 14 days . The canals were reinstrumented and irrigated with sterile saline solution . Sterile paper points were used to transfer the root canal contents to test tubes containing sterile saline solution . Part of the suspension was harvested in Sabouraud dextrose agar with chloramphenicol and incubated at 37 +/- 1 degree C for 48 h . CPMC was effective in 100% of the samples followed in decreasing order of effectiveness by calcium hydroxide with CPMC (70% effective), 1% sodium hypochlorite (70% effective) (p < 0.05), tricresol formalin (60% effective), 2% iodine-iodate solution (50% effective), calcium hydroxide paste (30% effective), and saline + no intracanal medication.

Boll Chim Farm, 2001 May-Jun, 140(3), 140 - 8
Synthesis and antimicrobial activity of novel pyrazole, pyrazoline, pyrazolinone and pyrazolidinedione derivatives of benzimidazole; Soliman R et al.; Four novel series of pyrazolylbenzimidazole derivatives have been prepared, namely 2-{(1-substituted phenyl-3,5-dimethyl-4-pyrazolyl)methyl}benzimidazole 5a-d 2-{(1-substituted phenyl-3-methyl-5-oxo-4,5-dihydro-4-pyrazolyl-4-yl)methyl}benzimidazoles 6a-d; 2-{(1-substituted phenyl-3,5-dioxopyrazolidin-4-yl)methyl}benzimidazoles 7a-d and 2-{(4-(1-phenyl-5-aryl-4,5-dihydro-3-pyrazolyl)phenylaminoacetyl}thio- methyl)-benzimidazoles 12a-e . The antimicrobial testing of the prepared compounds was performed using Escherichia Coli (NCTC 5933) as Gram-negative bacteria, Staphylococcus aureus (NCTC 4163) as gram-positive bacteria and Candida albicans (NCTC 5310) as yeast like fungi . The most potent compound was the pyrazolone 6a which exhibits interesting antibacterial activity against the gram-negative bacteria E . coli.

Biochem J, 2001 Aug 15, 358(Pt 1), 119 - 25
Selective induction of phospholipase D1 in pathogen-activated human monocytes; Locati M et al.; Phospholipase D (PLD) activation is part of the complex signalling cascade induced during phagocyte activation . Two PLD isoforms have been cloned, but their role in phagocyte functions is still poorly defined . We report that resting fresh circulating human monocytes expressed PLD1 . PLD1 protein expression was rapidly down-regulated during cell culture . Lipopolysaccharide and pathogen-derived agonists (Candida albicans, arabinoside-terminated lipoarabinomannan and Gram-positive bacteria, but not mannose-capped lipoarabinomannan or double-stranded RNA) strongly induced PLD1 expression at both the mRNA and protein levels . Pro-inflammatory cytokines {interleukin (IL)-1beta and tumour necrosis factor alpha} had only a weak effect, whereas immune cytokines (IL-6 and interferon gamma), anti-inflammatory cytokines (IL-13 and IL-10) and chemoattractants (fMet-Leu-Phe and macrophage chemoattractant protein 1) were inactive . None of the agonists tested induced significant changes in the basal expression of PLD2 mRNA . Consistent with PLD1 up-regulation was the observation that PLD enzymic activity was higher in monocytes treated with active-pathogen-derived agonists than in control cells, when stimulated with PMA or with chemotactic agonists (fMet-Leu-Phe and C5a) . Thus PLD2 seems to be a constitutive enzyme in circulating monocytes . Conversely, PLD1 is an inducible protein, rapidly regulated during culture conditions and selectively induced during cell activation . Therefore PLD1 might have a relevant role in immune responses against pathogens and in chronic inflammation.

Biotechnol Appl Biochem, 2001 Aug, 34(Pt 1), 47 - 54
Two telomerase reverse transcriptases (TERTs) expressed in Candida albicans; Metz AM et al.; The human pathogenic yeast Candida albicans contains two telomerase reverse transcriptase (TERT) genes . CaTERT1 and CaTERT2 appear either to be two alleles of the same gene or two entirely different genes that encode 867-residue proteins that differ by five amino acids . Both TERTs have a calculated pI of 9.5 and a M(r) of 100.9 kDa and are the smallest TERTs identified to date . Both genes appear to be expressed . Based on sequence similarity between CaTERT1 and the Saccharomyces cerevisiae orthologue Est2p, we suggest a revised alignment for motif E of Est2p . The identification of these TERT genes provides the first opportunity to study telomerase in an important human pathogen.

Yeast, 2001 Aug, 18(11), 1047 - 52
The centromere-binding factor Cbf1p from Candida albicans complements the methionine auxotrophic phenotype of Saccharomyces cerevisiae; Eck R et al.; A gene encoding the centromere binding factor 1 (Cbf1p) of the human pathogenic yeast Candida albicans was cloned and characterized . An open reading-frame was detected which encoded a 223 amino acid protein with a calculated molecular weight of 25.8 kDa and a relative isoelectric point of 5.55 . It shares 39% overall amino acid sequence identity with Saccharomyces cerevisiae Cbf1p . We localized the CaCBF1 gene on chromosome 4 . Southern analysis indicated that CaCBF1 is probably present as a single copy gene per haploid genome . The CaCBF1 gene under the control of its own promoter was able to complement the methionine auxotrophic growth, the increased mitotic instability of CEN plasmids and the slow growth of a Saccharomyces cerevisiae cbf1Delta mutant strain .

Yeast, 2001 Aug, 18(11), 1035 - 46
Extensive chromosome translocation in a clinical isolate showing the distinctive carbohydrate assimilation profile from a candidiasis patient; Iwaguchi SI et al.; Variation of the electrophoretic karyotype is common among clinical strains of Candida albicans and chromosome translocation is considered one of the causes of karyotypic variation . Such chromosome translocations may be a mechanism to confer phenotypic diversity on the imperfect fungus C . albicans . A clinical strain, TCH23, from a vaginal candidiasis patient shows distinct carbohydrate assimilation profile, serotype B, no chlamydospore formation and an atypical karyotype (Asakura et al., 1991) . To examine the taxonomic relationship among C . albicans, Candida dubliniensis and this strain, we sequenced the internal transcribed spacer 1 (ITS1) of nuclear ribosomal DNA . The ITS1 sequence of TCH23 was identical with that of C . albicans but not of C . dubliniensis . Thus, strain TCH23 was classified as a variant of C . albicans with an atypical phenotype . The chromosomal DNAs of this strain were resolved into 13 bands on pulse-field gel electrophoresis (PFGE) . Using DNA probes located at or near both ends of each chromosome of C . albicans, we investigated the chromosome organization of this strain . Referring to the SfiI map of C . albicans 1006 (Chu et al., 1993), we found that seven chromosomal DNA bands in strain TCH23 were reciprocal chromosome translocations . One homologue from chromosomes 1, 2 and 6 and both homologues from chromosomes 4 and 7 participated in these events . One translocation product was composed of three SfiI fragments, one each from chromosomes 2, 4 and 7 . We deduced the breakpoints of chromosome translocation from the physical map of this strain; between 1J and 1J1, between 2A and 2U, both ends of 4F2, between 6C and 6O and both ends of 7F .

Exp Lung Res, 2001 Jul-Aug, 27(5), 417 - 31
Role of intercellular adhesion molecule 1 in acute lung injury induced by candidemia; Yokomura I et al.; Candidemia, a complication often affecting immunocompromised patients, is a common cause of acute lung injury . Yeast-phase Candida albicans has been shown to express a protein that is antigenically and structurally related to Mac-1 . C . albicans is reported to stimulate intercellular adhesion molecule 1 (ICAM-1) expression on endothelial cells . In this study, the authors examined the role of ICAM-1 in acute lung injury induced by candidemia . The authors cultured rat pulmonary artery endothelial cells (RPAEC) and investigated the effect of anti-ICAM-1 antibodies on adhesion of C . albicans to RPAEC . In addition, the authors administered anti-ICAM-1 antibodies to rats to examine the effect of the antibodies on experimentally induced candidemia . Survival rates, lung wet-to-dry (W/D) weight ratios, bronchoalveolar lavage (BAL) fluid, histopathological findings, and colony-forming units (CFUs) of lung C . albicans were examined . The adherence of C . albicans to RPAEC was significantly decreased by anti-ICAM-1 antibodies . Anti-ICAM-1 antibodies significantly increased survival, decreased lung W/D weight ratios, decreased neutrophil counts in the BAL fluid, reduced microscopic lung injury, and decreased the quantity of lung C . albicans . These results indicate that ICAM-1 plays a role in adherence of C . albicans to pulmonary vascular endothelial cells, which likely leads to invasion of lung tissue by the organism.

Glycobiology, 2001 Aug, 11(8), 693 - 701
Peptides that mimic Candida albicans-derived beta-1,2-linked mannosides; Jouault T et al.; Beta-1,2-linked mannosides from Candida albicans phosphopeptidomannan (PPM) bind to macrophages through a receptor independent from the macrophage alpha-linked mannose receptor and stimulate these cells to secrete immune mediators . Anti-beta-1,2-linked mannoside but not anti-alpha-linked mannoside antibodies produced after immunization with neoglycoproteins protect animals from disseminated candidiasis . In this study, peptides that mimic beta-1,2-linked mannosides were isolated using phage display methodology . A phage library expressing random peptides was panned with an anti-beta-1,2-linked mannoside monoclonal antibody (mAb) . After three rounds of biopanning, the isolated phages were able to inhibit recognition of C . albicans by the mAb . Sixty percent of the phages had an identical DNA insert corresponding to the peptide sequence FHENWPS that was recognized specifically by the mAb . Injection of KLH-coupled peptide into mice generated high titers of polyclonal antibodies against C . albicans yeast cell walls . The anti-FHENWPS antibodies bound to C . albicans PPM and were inhibited by soluble beta-1,2-mannotetraose . Together, these data provide evidence for mimotopic activity of the peptide selected by biopanning with the anti-beta-1,2-oligomannoside mAb.

Biochim Biophys Acta, 2001 Aug 15, 1527(3), 141 - 8
Dicynthaurin: an antimicrobial peptide from hemocytes of the solitary tunicate, Halocynthia aurantium; Lee IH et al.; We isolated a novel antimicrobial peptide, dicynthaurin, from hemocytes of a tunicate, Halocynthia aurantium . The native peptide had a mass of approximately 6.2 kDa and was composed of two 30-residue monomers without sequence homology to any previously identified peptides (ILQKAVLDCLKAAGSSLSKAAITAIYNKIT) . Most cynthaurin molecules were C-terminally amidated and were linked covalently by a single cystine disulfide bond . When performed in membrane-mimetic environments, circular dichroism studies of dicynthaurin revealed largely alpha-helical conformations . Dicynthaurin's broad-spectrum activity encompassed Gram-positive (Micrococcus luteus, Staphylococcus aureus, Listeria monocytogenes) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), but not Candida albicans, a fungus . Although dicynthaurin was purified from a marine invertebrate, its antimicrobial activity was optimal at NaCl concentrations below 100 mM . This suggests that the antimicrobial actions of this molecule may take place intracellularly (e.g., within a phagosome) rather than extracellularly.

J Med Microbiol, 2001 Aug, 50(8), 743 - 7
Different isoforms of secreted aspartyl proteinases (Sap) are expressed by Candida albicans during oral and cutaneous candidosis in vivo; Schaller M et al.; Distinct isoforms of secreted aspartyl proteinases (Sap) of Candida albicans are important virulence factors for different types of candidosis . Predominant expression of Sap1-3 has been shown to be crucial for superficial infections in experimental mucosal and cutaneous candidosis, whereas Sap4-6 might be important for systemic disease . This in-vivo study investigated Sap expression in two samples from patients with oral candidosis and from cutaneous infection . Two different polyclonal antibodies directed against Sap1-3 and Sap4-6 were used for ultrastructural characterisation of protein localisation and expression . Post-embedding immuno-electron microscopy revealed Sap1-3 and Sap4-6 immunoreactivity in all samples . All C . albicans cells expressed predominantly the proteinases Sap1-3 which were evenly distributed within the cell wall and cytoplasmic membrane . In contrast, Sap4-6 labelling was only evident in a few fungal cells . In particular it was localised at the tips of hyphal cells during invasion . These data suggest a different pathogenetic role for Sap1-3 and Sap4-6 during host-fungal interaction.

Zhonghua Jie He He Hu Xi Za Zhi, 1998 Nov, 21(11), 661 - 3
{Studies on the biological and clinical characteristics of acquired pneumonia caused by K . planticola}; Luo G et al.; OBJECTIVE: To investigate the characteristics of biological, and clinical epidemiology of acquired pneumonia caused by a new type of Klebsiella, K . planticola . METHOD: 9 strains of K . planticola were isolated from respiratory samples of patients, in vitro, biological and serologic identification were done, in vivo, infected animal models were also evaluated . Clinical epidemiological inquiries were also performed . RESULT: 9 strains' biological features were different from those of the other Klebsiella . The 9 patients' ages were over forty and the primary disease was the third type tuberculosis of lungs . Using of antituberculosis drugs for long term as well as penicillins might be the predisposing factors, corticosteroids were used in 3 of the 9 patients, Combined infection with other pathogens in addition to K . planticola occurred in 8 patients, in 7/8, Candida albicans was identified . Drug sensitive tests show that all of them were resistant to penicillins and sensitive to the second and third generation cephalosporins . CONCLUSION: The new strain of Klebsiella was studied and this information will be useful for diagnosing and treating K . platicola.

FEMS Immunol Med Microbiol, 2001 Jul, 31(1), 65 - 71
HIV protease inhibitors attenuate adherence of Candida albicans to epithelial cells in vitro; Bektic J et al.; Oropharyngeal candidiasis is one of the first and most commonly reported opportunistic infections of untreated AIDS patients . With the introduction of the new antiviral HAART therapy, including HIV protease inhibitors, this mucocutaneous infection is nowadays only rarely observed in treated patients . It was recently shown that HIV protease inhibitors have a direct attenuating effect on Candida albicans secreted aspartic proteinases (Saps), an investigation prompted by the fact that both Sap and HIV protease belong to the superfamily of aspartic proteinases and by the observation that mucocutaneous infections sometimes resolve even in the absence of an immunological improvement of the host . As these Saps are important fungal virulence factors and play a key role in adhesion to human epithelial cells we tried to assess the effect of the HIV protease inhibitors Ritonavir, Indinavir and Saquinavir on fungal adhesion to these cells . The effect on phagocytosis by polymorphonuclear leukocytes was also assessed . Ritonavir was found to be the most potent inhibitor of fungal adhesion . A dose-dependent inhibition of adhesion to epithelial cells was found already at 0.8 microM and was significant at 4 microM or higher, at 500 microM the inhibition was about 55% . Indinavir and Saquinavir inhibited significantly at 4 microM or 20 microM, respectively; at 500 microM the inhibition was 30% or 50% . In contrast, no protease inhibitor was able to modulate phagocytosis of Candida by polymorphonuclear leukocytes . In conclusion, inhibition of Saps by HIV protease inhibitors may directly help to ease the resolution of mucosal candidiasis . In future, derivatives of HIV protease inhibitors, being more specific for the fungal Saps, may form an alternative in the treatment of mucosal candidiasis insensitive to currently available antimycotics.

Clin Pharmacokinet, 2001, 40(6), 441 - 72
Pharmacokinetics of antifungal agents in onychomycoses; Debruyne D et al.; Onychomycosis is caused by infection by fungi, mainly dermatophytes and nondermatophyte yeasts or moulds; it affects the fingernails and, more frequently, the toenails . Dermatophytes are responsible for about 90 to 95% of fungal infections . Trichophyton rubrum is the most common dermatophyte; Candida albicans is the major nondermatophyte yeast . Although topical therapy of onchomycosis does not lead to systemic adverse effects or interactions with concomitantly taken drugs, it does not provide high cure rates and requires complete compliance from the patient . At present there are 3 oral antifungal medications that are generally used for the short term treatment of onychomycosis: itraconazole, terbinafine and fluconazole . The persistence of these active drugs in nails allows weekly administration, reduced treatment or a pulse regimen . Good clinical and mycological efficacies are obtained with itraconazole 100 to 200 mg daily, terbinafine 250mg daily for 3 months, or fluconazole 150 mg weekly for at least 6 months . Itraconazole is a synthetic triazole with a broad spectrum of action . It is well absorbed when administered orally and can be detected in nails 1 to 2 weeks after the start of therapy . The nail : plasma ratio stabilises at around 1 by week 18 of treatment . Itraconazole is still detectable in nails 27 weeks after stopping administration . Nail concentrations are higher than the minimum inhibitory concentration (MIC) for most dermatophytes and Candida species from the first month of treatment . The elimination half-life of itraconazole from nails is long, ranging from 32 to 147 days . Terbinafine is a synthetic allylamine that is effective against dermatophytes . Terbinafine is well absorbed from the gastrointestinal tract, and the time to reach effective concentrations in nail is 1 to 2 weeks . The half-life is from 24 to 156 days, explaining the observed persistence of terbinafine in nails for longer than 252 days . Fluconazole is a bis-triazole broad spectrum antifungal with high oral bioavailability . The uptake of fluconazole by nail increases with the length of treatment, and nail : plasma ratios are generally 1.5 to 2 at steady state . Fluconazole concentrations exceed the MIC for Candida species soon after the start of treatment . Fluconazole concentrations fall slowly after the drug is stopped, with a half-life of 50 to 87 days, and fluconazole is still detectable in nails 5 months after the end of treatment . All these drugs are potent inhibitors of cytochrome P450 (CYP) enzymes and may increase the plasma concentrations of concomitantly used drugs . Itraconazole inhibits CYP3A4 . Fluconazole inhibits CYP3A4, but to a lesser degree than itraconazole, CYP2C9 and CYP2C19 . Terbinafine inhibits CYP2D6.

J Clin Microbiol, 2001 Aug, 39(8), 2999 - 3001
Serological differentiation of experimentally induced Candida dubliniensis and Candida albicans infections; Moragues MD et al.; Using a rabbit model of systemic infection, we show that it is possible to differentiate infections caused by Candida dubliniensis and other Candida species by detecting the antibody response mounted by the infected animals . These results confirm our previous observation in a patient with C . dubliniensis candidemia and suggest that detection of C . dubliniensis-specific antibodies is useful in the diagnosis of invasive candidiasis caused by this yeast.

J Clin Microbiol, 2001 Aug, 39(8), 2971 - 4
Candida species exhibit differential in vitro hemolytic activities; Luo G et al.; A total of 80 Candida isolates representing 14 species were examined for their respective responses to an in vitro hemolytic test . A modification of a previously described plate assay system where the yeasts are incubated on glucose (3%)-enriched sheep blood agar in a carbon dioxide (5%)-rich environment for 48 h was used to evaluate the hemolytic activity . A group of eight Candida species which included Candida albicans (15 isolates), C . dubliniensis (2), C . kefyr (2), C . krusei (4), C . zeylanoides (1), C . glabrata (34), C . tropicalis (5), and C . lusitaniae (2) demonstrated both alpha and beta hemolysis at 48 h postinoculation . Only alpha hemolysis was detectable in four Candida species, viz., C . famata (3), C . guilliermondii (4), C . rugosa (1), and C . utilis (1), while C . parapsilosis (5) and C . pelliculosa (1) failed to demonstrate any hemolytic activity after incubation for 48 h or longer . This is the first study to demonstrate the variable expression profiles of hemolysins by different Candida species.

J Nat Prod, 2001 Jul, 64(7), 965 - 7
Multiplolides A and B, new antifungal 10-membered lactones from Xylaria multiplex; Boonphong S et al.; Two new 10-membered lactones, namely, multiplolides A (1) and B (2), were isolated from the broth extract of the fungus Xylaria multiplex BCC 1111 . Chemical structures of 1 and 2 were elucidated on the basis of their spectral data . Multiplolides A (1) and B (2) exhibited antifungal activity against Candida albicans with IC(50) values of 7 and 2 microg/mL, respectively . Both 1 and 2 were inactive in the screening systems toward the malarial parasite Plasmodium falciparum (at 20 microg/mL) and were not cytotoxic to BC-1 and KB cell lines (at 20 microg/mL).

J Endod, 2000 Dec, 26(12), 695 - 8
Occurrence of Candida albicans in infections of endodontic origin; Baumgartner JC et al.; Microorganisms are recognized as the etiological agent for the majority of pulpal and periradicular disease . Although bacteria have been the most studied, fungi have also been associated with infected root canals . The purpose of this study was to evaluate the contents of infected root canals and aspirates of cellulitis/abscesses of endodontic origin for the presence of Candida albicans using the polymerase chain reaction (PCR) . PCR primers specific for the 18S ribosomal RNA gene of C . albicans were used to survey 24 samples taken from infected root canals and 19 aspirates from periradicular infections of endodontic origins . The presence of C . albicans was detected in 5 of 24 (21%) samples taken from root canals, but none was detected in the periradicular aspirates . The results indicate that PCR is an extremely sensitive molecular method that may be used to identify C . albicans directly in samples from infections of endodontic origin.

Gene, 2001 Jul 11, 272(1-2), 157 - 64
A DNA polymorphism specific to Candida albicans strains exceptionally successful as human pathogens; Giblin L et al.; A large proportion of infection-causing isolates of the yeast Candida albicans belong to a general-purpose genotype, identifiable by fingerprinting with the moderately repetitive sequence Ca3 . The high prevalence of this group -- up to 70% in some patient categories -- suggests that its members possess genetic determinants, which enhance their success as pathogens compared to other strains . To find such determinants we are comparing the genomes of representatives of the general-purpose genotype cluster with the genomes of other strains . In this paper we describe the identification of a 985 bp HpaII fragment (MU13-4) specific to general-purpose genotype strains . The fragment was present in 90% of these strains, but only in 10% of other strains . The fragment did not hybridize with probe Ca3, used to define the general-purpose cluster . It contains elevated levels of repetitive DNA . Sequences homologous to MU13-4 are dispersed throughout the chromosomes of general-purpose strains but are rarer or absent in other strains, as judged by Southern hybridization . Using the Stanford C . albicans genome database, we have placed the MU13-4 fragment next to a CARE-1 element . We also found 79 significant homologies between parts of MU13-4 and 19 other contigs . Attempts to amplify the region surrounding the polymorphic fragment in non-general-purpose genotype strains suggest, as do the hybridization data, that the polymorphism is created by a deletion in non-cluster strains . These results show that it is possible to identify polymorphisms specific to general-purpose genotype strains . Primers against the fragment will allow PCR-based discrimination between general-purpose genotype strains and other strains, facilitating investigations aimed at determining morbidity and mortality caused by general-purpose genotype strains compared to other strains.

Curr Pharm Biotechnol, 2000 Nov, 1(3), 235 - 51
Cytokines in candidiasis and aspergillosis; Mencacci A et al.; Both innate and T helper (Th) immunity play a central role in fungal infections . A bi-directional influence exists between the two compartments of the immune system, mainly occurring through cytokine production . On the one hand, protective Th1 or nonprotective Th2 cells mediate resistance or susceptibility to disseminated and localized fungal infections by secreting cytokines with activating or deactivating signals for effector phagocytic cells . On the other hand, cells of the innate immune system regulate the development of antifungal T helper responses by producing directive cytokines, such as interleukin (IL)-12 and IL-10 . In experimental models of Candida albicans and Aspergillus fumigatus infections, the administration or neutralization of selective cytokines and the use of cytokine-deficient mice have revealed the existence of a hierarchical pattern of cytokine mediated regulation of antifungal Th cell development and effector function . A finely regulated balance of directive cytokines, rather than the relative absence of opposing cytokines, appears to be required for optimal development and maintenance of protective Th1 reactivity to fungi . Thus, it is conceivable that some cytokines may have beneficial or deleterious effects on infection, depending on the dose and timing of endogenous production or exogenous administration . A better understanding of the different, sometimes unexpected, roles of cytokines is required for their use in prophylaxis and therapy of fungal infections, either alone or in combination with antifungal agents.

J Immunol, 2001 Aug 1, 167(3), 1550 - 7
Complement is essential for protection by an IgM and an IgG3 monoclonal antibody against experimental, hematogenously disseminated candidiasis; Han Y et al.; The incidence of life-threatening, hematogenously disseminated candidiasis, which is predominantly caused by Candida albicans, parallels the use of modern medical procedures that adversely affect the immune system . Limited antifungal drug choices and emergence of drug-resistant C . albicans strains indicate the need for novel prevention and therapeutic strategies . We are developing vaccines and Abs that enhance resistance against experimental candidiasis . However, the prevalence of serum anti-Candida Abs in candidiasis patients has led to the misconception that Abs are not protective . To explain the apparent discrepancy between such clinical observations and our work, we compared functional activities of C . albicans-specific protective and nonprotective mAbs . Both kinds of Abs are agglutinins that fix complement and are specific for cell surface mannan, but the protective Abs recognize beta-mannan, and the nonprotective Ab is specific for alpha-mannan . By several indirect and direct measures, the protective mAbs more efficiently bind complement factor C3 to the yeast cell than do nonprotective Ab . We hypothesize that the C3 deposition causes preferential association of blood-borne fungi with host phagocytic cells that are capable of killing the fungus . We conclude from these results that the protective potential of Abs is dependent on epitope specificity, serum titer, and ability to rapidly and efficiently fix complement to the fungal surface . The mechanism of protection appears to be associated with enhanced phagocytosis and killing of the fungus.

Ann Dermatol Venereol, 2001 Jun-Jul, 128(6-7), 733 - 6
{Onychomycosis in Tunis area: epidemiological and mycological data}; Anane S et al.; BACKGROUND: Onychomycosis is by far the most frequent cause of nail disease . We describe epidemiological and mycological features observed in the Tunis area in Tunisia . MATERIAL AND METHODS: Data were collected from 292 nail samples performed in 255 patients with suspected onychomycosis . RESULTS: Request for samples were made late, on the average 48 months after development of nail disorders . Most of the patients were women (63.5%) . One hundred ninety-six samples were positive (67%), 130 from toe nails and 66 from finger nails . Simultaneous infections of both finger and toe nails were found in 22 cases . Associated onychomycosis and skin mycosis was found preferentially in feet onychomycosis . The sensitivities of direct examination and culture depended on the site of the onychomycosis . Cultures were more sensitive for hands where yeasts, particularly Candida albicans, predominated, but the direct examination was more sensitive for feet where dermatophytes, particularly Trichophyton rubrum, predominated . CONCLUSION: Mycological examination is compulsory for confirmation of onychomycosis . It is also recommended before initiating a costly long-term treatment.

Bioorg Med Chem Lett, 2001 Jul 23, 11(14), 1833 - 7
Design and synthesis of novel benzofurans as a new class of antifungal agents targeting fungal N-myristoyltransferase . Part 1; Masubuchi M et al.; Potent and selective Candida albicans N-myristoyltransferase (CaNmt) inhibitors have been identified through optimization of a lead compound 1 discovered by random screening . The inhibitor design is based on the crystal structure of the CaNmt complex with compound (S)-3 and structure-activity relationships (SARs) have been clarified . Modification of the C-4 side chain of 1 has led to the discovery of a potent and selective CaNmt inhibitor 11 (RO-09-4609), which exhibits antifungal activity against C . albicans in vitro.

J Oral Pathol Med, 2001 Jul, 30(6), 336 - 46
Heterogeneity in antifungal susceptibility of clones of Candida albicans isolated on single and sequential visits from a HIV-infected southern Chinese cohort; Samaranayake YH et al.; The increased frequency and severity of candidal infections in human immunodeficiency virus (HIV)-infected individuals has prompted the wide use of antifungals, such as amphotericin B, ketoconazole, and fluconazole, resulting in the emergence of drug-resistant strains of Candida albicans . To study this phenomenon in an ethnic Chinese cohort, we isolated multiple colonies of Candida from the oral cavities of 16 HIV-infected patients on single and subsequent sequential visits over a period of 12 months . Ten of the 16 patients had sporadic episodes of oropharyngeal candidiasis (Group A), while the remainder were asymptomatic with respect to this condition (Group B) . Oral rinses were collected and immediately processed in the laboratory for the isolation of C . albicans in a standard manner . A total of 433 C . albicans isolates were tested for their susceptibility to amphotericin B, ketoconazole and fluconazole by an agar diffusion method using the commercially available E-test . All tested isolates demonstrated variable susceptibility to amphotericin B, ketoconazole and fluconazole . The minimum inhibitory concentration (MIC) of the isolates for amphotericin B, ketoconazole and fluconazole ranged from <0.002-1.5 microg/ml, <0.002-4.0 microg/ml and <0.016-32 microg/ml, respectively . Sequential isolates of a few patients demonstrated variable susceptibility to all the antifungals, and no discernible MIC pattern emerged either in group A or B over time . Interestingly, significant variation in antifungal susceptibility was also noted in isolates obtained from the same patient on a single visit . Sequential yeast isolates in 9 of 16 patients (56%) demonstrated significant differences in MIC within and between visits for both amphotericin B and ketoconazole, while a lower percentage--44%(7/16)--exhibited this trait for fluconazole . Our study demonstrates the diversity in antifungal susceptibility in either commensal or "infective" oral strains of C . albicans in HIV disease, and shows the need for vigilance for the emergence of resistant strains, and for frequent antifungal susceptibility studies.

Chemosphere, 2001 Jul, 44(3), 313 - 9
Study of fungicidal and antibacterial effect of the Cu(II)-complexes of thiophene oligomers synthesized in ZSM-5 zeolite channels; Cik G et al.; The influence of the Cu(II)-complexes of thiophene oligomers synthesized by oxidative polymerization of thiophene with Cu2+ ions in ZSM-5 zeolite channels on fungicidal and antimicrobial properties was studied . It has been found that the heterogeneous system culture medium-modified zeolite increases sporulation of the tested fungus (Aspergillus niger) and concurrently kills yeast (Candida albicans) . These effects are attributed to a slow release of Cu2+ ions and thiophene oligomers into the culture medium . As for the tested bacteria (G+ Staphylococcus aureus, G- Escherichia coli), the percentage of the killed cells increases due to light activation of the system . The light effect is assigned to photogeneration of the reactive oxygen species (ROS), mainly *OH radicals, which were registered in the water solution by EPR spectroscopy . It has been confirmed that the thiophene oligomers present in the Cu-ZSM-5 microstructure slow down the release of copper into the medium.

Zhonghua Yi Xue Za Zhi (Taipei), 2001 Apr, 64(4), 223 - 30
Risk factors of catheter-related infections in total parenteral nutrition catheterization; Wang FD et al.; BACKGROUND: The use of central venous catheter for administration of total parenteral nutrition (TPN) is a risk factor of catheter-related infections (CRIs) that are associated with increased morbidity and mortality, prolonged hospitalization, and increased medical costs . The purpose of this study is to evaluate the risk factors of CRIs in patients with administration of TPN . METHODS: A total of 1134 patients receiving TPN between January, 1996 and December, 1998 were studied . The category of infection included definite catheter-related bloodstream infection (CR-BSI), probable CR-BSI, and insertion site infection . Statistical analysis of risk factors was performed . RESULTS: A total of 131 episodes of CRI occurred, representing an infection rate of 11.46% . Ninety-three episodes (8.1%) had probable CR-BSI, 13 episodes (1.1%) had definite CR-BSI, and 25 episodes (2.2%) had insertion site infection . Duration of TPN infusion and frequency of catheter insertion showed statistically significant difference by logistic regression multivariate analysis . The isolated organisms were in sequence of coagulase-negative Staphylococci (19.4%), Staphylococcus aureus (17.2%) and Candida albicans (14.4%) . CONCLUSIONS: Risk factors influencing the occurrence of CRI in TPN administration were multifactorial; however, duration of TPN infusion and frequency of catheter insertion were the main factors in our study.

J Pharm Sci, 2001 Jul, 90(7), 902 - 14
Formulation and characterization of amphotericin B-polyethylenimine-dextran sulfate nanoparticles; Tiyaboonchai W et al.; A new aqueous nanoparticle system has been developed using complex coacervation employing the oppositely charged polymers polyethylenimine (PEI) and dextran sulfate (DS), with zinc sulfate as a stabilizing agent . Amphotericin B (AmB) was loaded into the nanoparticles as a model drug . The nanoparticles contained PEI and DS in the weight ratio of approximately 1:2 . They possessed a zeta potential of approximately +30 mV and demonstrated a narrow size distribution in the range 100-600 nm with a polydispersity index of 0.2 . Electron microscopy revealed spherical nanocapsules with a smooth surface . Very favorable drug entrapment and recovery efficiencies of up to 85% were routinely observed . Processing parameters, such as the pH of the PEI solutions, ratio of the two polymers, as well as the concentrations of DS and zinc sulfate, all played a significant role in controlling particle size . Dissolution studies demonstrated a fast release that is dependent on the model drug solubility . The AmB-loaded nanoparticles displayed no toxicity in tissue culture in contrast to free drug and were almost as efficacious as free drug in killing Candida albicans . Advantages of this simple technique are (1) ease of manufacturing and mild preparation conditions, (2) employment of completely aqueous processing conditions, (3) use of biocompatible polymers that can be prepared aseptically, (4) ability to control their size, and (5) a high level of drug entrapment.

Fungal Genet Biol, 2001 Jul, 33(2), 137 - 43
Regions of microsynteny in Magnaporthe grisea and Neurospora crassa; Hamer L et al.; A bacterial artificial chromosome (BAC) clone containing 110,467 bp of genomic DNA from Magnaporthe grisea was sequenced, annotated, and compared to the genomes of Neurospora crassa, Candida albicans, and Saccharomyces cerevisiae . Twenty-six open reading frames (ORFs), involved in multiple biochemical pathways, were identified in the BAC sequence . A region of 53 kb, containing 18 of the 26 ORFs, was found to be syntenic to a portion of the N . crassa genome . Subregions of complete colinearity as well as interrupted colinearity were present . No synteny was evident with either C . albicans or S . cerevisiae . The identification of syntenic regions containing highly conserved genes across two genera that have been evolutionarily separated for approximately 200 million years elicits many biological questions as to the function and identity of these genes .

Chem Pharm Bull (Tokyo), 2001 Jul, 49(7), 927 - 9
Diastereomeric diterpenes from Coleus blumei; Ragas CY et al.; The chloroform extract of the air-dried leaves of Coleus blumei afforded a mixture of diastereomers of a new abietane type diterpene whose structures were elucidated by extensive one and two dimensional (ID, 2D) NMR and mass spectrometry . Acetylation of the mixture afforded a single compound . Antimicrobial tests on the diterpene indicate that it is active against Bacillus subtilis, Pseudomonas aeruginosa and Candida albicans.

Mol Microbiol, 2001 Jul, 41(1), 19 - 31
The germ tubes of Candida albicans hyphae and pseudohyphae show different patterns of septin ring localization; Sudbery PE; The location of the septin ring in the germ tubes of Candida albicans hyphae and pseudohyphae was studied using an antibody to Saccharomyces cerevisiae Cdc11p . In pseudohyphae induced by growth at 35 degrees C in YEPD or Lee's medium, a septin ring formed at or near (mean 1.8 microm) the neck between the mother cell and the germ tube . This became double later in the cycle, and the first mitosis took place across the plane of this double ring . A septin ring also formed at the germ tube neck of developing hyphae induced by serum or growth on Lee's medium at 37 degrees C . However, at later times, this ring became disorganized and disappeared . A second double ring then appeared 10-15 microm (mean 12.5 microm) along the length of the germ tube . The nucleus subsequently migrated out of the mother cell into the germ tube, and the first mitosis took place across the plane of this second septin ring . The relocation of the septin ring in developing hyphae provides a clear-cut molecular distinction between hyphae and pseudohyphae . Commitment to one type of septin localization and mitosis was shown to occur early in the first mitotic cycle, well before evagination . Germ tubes of hyphae and pseudohyphae also have different widths . A point of commitment to germ tube width was also demonstrated, but occurred later in the cycle, approximately coincident with the time of evagination.

Nature, 2001 Jul 5, 412(6842), 83 - 6
The glyoxylate cycle is required for fungal virulence; Lorenz MC et al.; Candida albicans, a normal component of the mammalian gastrointestinal flora, is responsible for most fungal infections in immunosuppressed patients . Candida is normally phagocytosed by macrophages and neutrophils, which secrete cytokines and induce hyphal development in this fungus . Neutropenic patients, deficient in these immune cells, are particularly susceptible to systemic candidiasis . Here we use genome-wide expression profiles of the related yeast Saccharomyces cerevisiae to obtain a signature of the events that take place in the fungus on ingestion by a mammalian macrophage . Live S . cerevisiae cells isolated from the phagolysosome are induced for genes of the glyoxylate cycle, a metabolic pathway that permits the use of two-carbon compounds as carbon sources . In C . albicans, phagocytosis also upregulates the principal enzymes of the glyoxylate cycle, isocitrate lyase (ICL1) and malate synthase (MLS1) . Candida albicans mutants lacking ICL1 are markedly less virulent in mice than the wild type . These findings in fungi, in conjunction with reports that isocitrate lyase is both upregulated and required for the virulence of Mycobacterium tuberculosis, demonstrate the wide-ranging significance of the glyoxylate cycle in microbial pathogenesis.

J Biol Chem, 2001 Sep 21, 276(38), 35875 - 82 Epub 2001 Jul 12.
Structural and biological characterization of chromofungin, the antifungal chromogranin A-(47-66)-derived peptide; Lugardon K et al.; Vasostatin-I, the natural fragment of chromogranin A-(1-76), is a neuropeptide able to kill a large variety of fungi and yeast cells in the micromolar range . We have examined the antifungal properties of synthetic vasostatin-I-related peptides . The most active shortest peptide, named chromofungin, corresponds to the sequence Arg(47)-Leu(66) . Extensive (1)H NMR analysis revealed that it adopts a helical structure . The biophysical mechanism implicated in the interaction of chromofungin with fungi and yeast cells was studied, showing the penetration of this peptide with different lipid monolayers . In order to examine thoroughly the antifungal activity of chromofungin, confocal laser microscopy was used to demonstrate the ability of the rhodamine-labeled peptide to interact with the fungal cell wall, to cross the plasma membrane, and to accumulate in Aspergillus fumigatus, Alternaria brassicola, and Candida albicans . Our present data reveal that chromofungin inhibits calcineurin activity, extending a previous observation that the N-terminal region of chromogranin A interacts with calmodulin in the presence of calcium . Therefore, the destabilization of fungal wall and plasma membrane, together with the possible intracellular inhibition of calmodulin-dependent enzymes, is likely to represent the mechanism by which vasostatin-I and chromofungin exert antifungal activity.

J Eur Acad Dermatol Venereol, 2001 Jan, 15(1), 34 - 8
Systemic ketoconazole for yeast allergic patients with atopic dermatitis; Back et al.; BACKGROUND: Adult patients with atopic dermatitis (AD), especially with the head-neck distribution, are often sensitized to the lipophilic yeast Malassezia furfur/Pityrosporum orbiculare, which is considered to contribute to the pathogenesis of the dermatitis . OBJECTIVE: Evaluation of the efficacy of oral ketoconazole on immunological and clinical parameters in yeast allergic adult patients with AD . STUDY DESIGN: Randomized double-blind placebo-controlled study . SUBJECTS: Twenty-nine patients with specific IgE antibodies to M . furfur/P . orbiculare and with elevated serum IgE participated in the investigation . Fifteen subjects were treated with 200 mg ketoconazole daily and 14 received placebo for 3 months . Betamethasone cream was allowed as supplementary therapy and the consumption was registered . The clinical score (SCORAD), total serum IgE and specific IgE antibodies to M . furfur/P . orbiculare, Candida albicans and Dermatophagoides pteronyssinus were monitored at the starting point and by the end of the first and third month . RESULTS: In the actively treated group the levels of specific IgE to M . furfur/P . orbiculare and C . albicans as well as total serum IgE decreased significantly . Sensitization to D . pteronyssinus was not influenced . The clinical score decreased in both groups, and the improvement was correlated to the consumption of topical steroids in the control group but not in the ketoconazole group.

Fitoterapia, 2000 Sep, 71(5), 570 - 3
Antimicrobial activity of Fabaceae species used in Yucatan traditional medicine; Rosado-Vallado M et al.; The methanol and water extracts of six Fabaceae species, traditionally used in Mayan medicine for the treatment of diarrhoea and eye infections, were phytochemically screened and tested for in vitro antimicrobial activity . Four species showed activity against Gram positive bacteria, five exhibited some activity against Candida albicans, two exhibited activity against Aspergillus niger and only one, Mimosa pigra, inhibited growth of Pseudomonas aeruginosa . None of the extracts was active against Escherichia coli.

Yeast, 2001 Jul, 18(10), 915 - 22
Molecular cloning of CaYRB1, the Candida albicans RanBP1/YRB1 homologue; Clement M et al.; The yeast Ran binding protein 1 (Yrb1p) is a small protein of 23 kDa that is highly conserved among eukaryotes . It stimulates the GTPase activity of Gsp1p in the presence of the GTPase activating protein Rna1p . In addition to its role in nucleocytoplasmic transport of macromolecules, YRB1/RanBP1 could be involved in the regulation of microtubules structure and dynamics . Since microtubules are tightly associated with morphological changes, we have been interested to study the role and function of YRB1 in the pathogenic fungus Candida albicans, where there is regulated change in cellular morphology . The gene product of CaYRB1 encodes a 212 amino acid protein displaying 73% homology to the S . cerevisiae homologue . The bacterially expressed gene product has an apparent molecular weight of 35.7 kDa . We show that it can complement a S . cerevisiae yrb1 null mutant and that its mRNA does not appear to be regulated in response to conditions inducing morphological changes in C . albicans .

Infect Immun, 2001 Aug, 69(8), 5115 - 20
Invasive candidiasis stimulates hepatocyte and monocyte production of active transforming growth factor beta; Letterio JJ et al.; Candida albicans is an opportunistic fungal pathogen and a major cause of morbidity and mortality in patients with compromised immune function . The cytokine response to tissue invasion by C . albicans can influence the differentiation and function of lymphocytes and other mononuclear cells that are critical components of the host response . While the production of transforming growth factor beta (TGF-beta) has been documented in mice infected with C . albicans and is known to suppress phagocyte function, the cellular source and role of this cytokine in the pathogenesis of systemic candidiasis are not well understood . We have investigated the source of production of TGF-beta by immunohistochemical studies in tissue samples from patients with an uncommon complication of lymphoreticular malignancy, chronic disseminated candidiasis (CDC), and from a neutropenic-rabbit model of CDC . Liver biopsy specimens from patients with documented CDC demonstrated intense staining for extracellular matrix-associated TGF-beta1 within inflammatory granulomas, as well as staining for TGF-beta1 and TGF-beta3 within adjacent hepatocytes . These results correlate with the immunolocalization of TGF-beta observed in livers of infected neutropenic rabbits, using a neutralizing antibody that recognizes the mature TGF-beta protein . Human peripheral blood monocytes incubated with C . albicans in vitro release large amounts of biologically active TGF-beta1 . The data demonstrate that local production of active TGF-betas by hepatocytes and by infected mononuclear cells is a component of the response to C . albicans infection that most probably contributes to disease progression in the immunocompromised host.

Infect Immun, 2001 Aug, 69(8), 5046 - 55
Comparison of pathogenesis and host immune responses to Candida glabrata and Candida albicans in systemically infected immunocompetent mice; Brieland J et al.; Cytokine-mediated host defense against Candida glabrata infection was compared to that against C . albicans, using immunocompetent murine models of systemic candidiasis . The pathogenesis of infection was evaluated morphologically and by culture of target organs, while the kinetics of induction of cytokine mRNAs and corresponding proteins were determined in kidneys by real-time reverse transcription-PCR and cytokine-specific murine enzyme-linked immunosorbent assays, respectively . Systemic infection with C . glabrata resulted in a chronic, nonfatal infection with recovery of organisms from kidneys, while intravenous inoculation with C . albicans resulted in rapid mortality with logarithmic growth of organisms in kidneys and recovery of C . albicans from the spleen, liver, and lungs . Survival of C . glabrata-infected mice was associated with rapid induction of mRNAs and corresponding immunoreactive proteins for the proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha), interleukin-12 (IL-12), and gamma interferon (IFN-gamma) and the lack of induction of protein for the anti-inflammatory cytokine IL-10 . In contrast, mortality in C . albicans-infected mice was associated with induction of mRNA and corresponding protein for IL-10 but delayed (i.e., TNF-alpha) or absent (i.e., IL-12 and IFN-gamma) induction of immunoreactive proinflammatory cytokines . Mice were subsequently treated with cytokine-specific neutralizing monoclonal antibodies (MAbs) to TNF-alpha, IL-12, or IFN-gamma, and the effect on growth of C . glabrata in kidneys was assessed . Neutralization of endogenous TNF-alpha resulted in a significant increase in C . glabrata organisms compared to similarly infected mice administered an isotype-matched control MAb, while neutralization of endogenous IL-12 or IFN-gamma had no significant effect on C . glabrata replication . These results demonstrate that in response to intravenous inoculation of C . glabrata, immunocompetent mice develop chronic nonfatal renal infections which are associated with rapid induction of the proinflammatory cytokines TNF-alpha, IL-12, and IFN-gamma . Furthermore, TNF-alpha plays a key role in host defense against systemic candidiasis caused by either C . glabrata or C . albicans, as the absence of endogenous TNF-alpha activity was associated with enhanced tissue burden in both infection models.

Med Mycol, 2001 Jun, 39(3), 299 - 302
Aspergillus fumigatus CYP51 sequence: potential basis for fluconazole resistance; Edlind TD et al.; We have cloned and sequenced the Aspergillus fumigatus CYP51 gene which encodes the target of azole antifungal agents, namely cytochrome P450 sterol 14alpha-demethylase . Since A . fumigatus is intrinsically resistant to the widely used azole fluconazole, we compared its predicted CYP51 sequence to the CYP51 sequences from fluconazole-susceptible and resistant Candida albicans . This analysis generated specific hypotheses regarding the basis for A . fumigatus fluconazole resistance; in particular, A . fumigatus residue Ile301 corresponds to C . albicans residue Thr315 which is mutated to Ala in resistant strains and is proposed to hydrogen bond with the sterol substrate.

Med Mycol, 2001 Jun, 39(3), 283 - 5
Inhibition of hyphal growth of Candida albicans by activated lansoprazole, a novel benzimidazole proton pump inhibitor; Biswas SK et al.; The effect of activated lansoprazole (AG 2000), a novel benzimidazole proton pump inhibitor, against hypha formation of Candida albicans was examined in hypha-forming medium pH 7 (HFM7) after 20 h . AG 2000, at 50-800 microM, did not inhibit germ tube formation . However, it inhibited elongation of germ tubes to form hyphae and favored conversion of germ tubes to resume yeast growth at concentrations of > or =200 microM . Pre-treatment of AG 2000 with a sulfhydryl reagent (1:1), such as 2-mercaptoethanol . blocked the inhibitory property of AG 2000 on hypha formation.

Med Mycol, 2001 Jun, 39(3), 261 - 8
Oroesophageal candidiasis is lethal for transgenic mice with combined natural killer and T-cell defects; Balish E et al.; Germfree transgenic epsilon 26 (Tgepsilon26) mice, which express the full-length human CD3epsilon gene, have combined defects in natural killer (NK) cells and T cells were found to be extremely susceptible to oroesophageal (palate, tongue, esophagus) and gastric (cardia-antrum section) candidiasis . The gnotobiotic Tgepsilon26 mice die, apparently from severe oroesophageal candidiasis, within 2-4 weeks after their alimentary tracts are colonized with Candida albicans . The Tgepsilon26 mice manifest resistance to acute systemic candidiasis (intravenous injection) and to systemic candidiasis of endogenous origin for the first 2 weeks after their alimentary tracts are colonized with C . albicans . Granulocyte depletion data suggest that granulocytes, in the absence of functional NK cells and T cells, can protect Tgepsilon26 mice from acute systemic candidiasis and from systemic candidiasis of endogenous origin, for at least 14 days after alimentary tract colonization . Granulocytes and macrophages, in the absence of NK cells and T cells, are unable to protect Tgepsilon26 mice from lethal oroesophageal candidiasis and systemic candidiasis of endogenous origin which was evident in moribund Tgepsilon26 mice 2-4 weeks after colonization . Thus, non-T cells (i.e., NK cells) and T cells play important roles in resistance to oroesophageal and systemic (acute and of endogenous origin) candidiasis.

J Bacteriol, 2001 Aug, 183(15), 4614 - 25
The histone deacetylase genes HDA1 and RPD3 play distinct roles in regulation of high-frequency phenotypic switching in Candida albicans; Srikantha T et al.; Five histone deacetylase genes (HDA1, RPD3, HOS1, HOS2, and HOS3) have been cloned from Candida albicans and characterized . Sequence analysis and comparison with 17 additional deacetylases resulted in a phylogenetic tree composed of three major groups . Transcription of the deacetylases HDA1 and RPD3 is down-regulated in the opaque phase of the white-opaque transition in strain WO-1 . HOS3 is selectively transcribed as a 2.5-kb transcript in the white phase and as a less-abundant 2.3-kb transcript in the opaque phase . HDA1 and RPD3 were independently deleted in strain WO-1, and both switching between the white and opaque phases and the downstream regulation of phase-specific genes were analyzed . Deletion of HDA1 resulted in an increase in the frequency of switching from the white phase to the opaque phase, but had no effect on the frequency of switching from the opaque phase to the white phase . Deletion of RPD3 resulted in an increase in the frequency of switching in both directions . Deletion of HDA1 resulted in reduced white-phase-specific expression of the EFG1 3.2-kb transcript, but had no significant effect on white-phase-specific expression of WH11 or opaque-phase-specific expression of OP4, SAP1, and SAP3 . Deletion of RPD3 resulted in reduced opaque-phase-specific expression of OP4, SAP1, and SAP3 and a slight reduction of white-phase-specific expression of WH11 and 3.2-kb EFG1 . Deletion of neither HDA1 nor RPD3 affected the high level of white-phase expression and the low level of opaque-phase expression of the MADS box protein gene MCM1, which has been implicated in the regulation of opaque-phase-specific gene expression . In addition, there was no effect on the phase-regulated levels of expression of the other deacetylase genes . These results demonstrate that the two deacetylase genes HDA1 and RPD3 play distinct roles in the suppression of switching, that the two play distinct and selective roles in the regulation of phase-specific genes, and that the deacetylases are in turn regulated by switching.

Oral Microbiol Immunol, 2001 Aug, 16(4), 243 - 9
Increase of Candida cell virulence by anticancer drugs and irradiation; Ueta E et al.; The influence of anticancer drugs and irradiation on Candida cell proliferation, adherence to HeLa cells and susceptibility to antifungal drugs (amphotericin B and miconazole) and neutrophils were examined using two Candida albicans strains . After treatment with 5-fluorouracil (25 microg/ml to 250 microg/ml), cis-diammine-dichloroplatinum (10 microg/ml to 100 microg/ml), peplomycin (0.5 microg/ml to 5 microg/ml) or 137Cs (20 Gy to 40 Gy) for 3 days or more, surviving Candida cells proliferated more rapidly than did untreated control cells . Anticancer agent-pretreated Candida cells revealed an increased adhesion to HeLa cells corresponding to an increase of binding to the lectins . The concentration of half limited colony formation (IC50) of amphotericin B and miconazole was increased to near two-fold that of the control by pretreatment of Candida cells with the anticancer agents, except peplomycin, which only weakly increased IC50 . In addition, the enolase and Candida acid proteinase activities in the culture supernatants were increased by pretreatment with the drugs and irradiation . Correspondingly, surviving Candida cells after these treatments were resistant to neutrophils, with a reduction to half of the killing . These results indicate that anti-cancer drugs and irradiation potentiate the virulence of Candida cells, or they eliminate Candida cells with low virulence, thereby enhancing the risk of oral and systemic candidiasis.

J Small Anim Pract, 2001 Jun, 42(6), 291 - 7
Malassezia and Candida infections in bull terriers with lethal acrodermatitis; McEwan NA; In 12 cases of lethal acrodermatitis (LAD), four sampling techniques (brush, swab, scrape and adhesive tape strip) were used to study the distribution of yeasts in various body sites and these results were compared with those from five cases of atopic dermatitis and those of 10 normal dogs . Malassezia was frequently isolated from lesional and non-lesional skin and haircoat, footpads, nails and mucous membranes from dogs with either LAD or atopic dermatitis, although, generally, more Malassezia organisms were isolated from LAD cases . In normal dogs, Malassezia was most frequently recovered from the ear canal and the perianal skin . Candida was isolated frequently from dogs with LAD, but only a single isolate of this yeast was found in the other two groups . Fungal hyphae and pseudohyphae, probably Candida albicans, could be detected in samples collected from the nails and footpads of dogs with LAD . Both Malassezia and Candida could be isolated using all four sampling techniques . The MacKenzie (toothbrush) technique and adhesive tape strip cultures proved simple methods for the semiquantitative evaluation of yeasts . The high recovery rate of Malassezia and Candida from dogs with LAD is probably related to immune dysfunction, particularly T-cell dysfunction, known to be present in these dogs . C albicans infection may in part be responsible for the pathogenic changes of the nails and footpads commonly seen in cases of LAD.

Int J Med Microbiol, 2001 May, 291(2), 183 - 8
Gene regulation and host adaptation mechanisms in Candida albicans; Staib P et al.; The yeast Candida albicans is a harmless member of the normal microflora on the mucosal surfaces of most healthy persons, but it can cause severe opportunistic infections in immunosuppressed patients . To become a successful human commensal and pathogen, C . albicans has evolved host adaptation mechanisms on different levels . The regulated expression of virulence and other genes in response to environmental signals allows an optimal adaptation to new host niches during the course of an infection . In addition, C . albicans is able to switch between different cell types in a reversible and apparently random fashion . Phenotypic switching involves the coordinated regulation of phase-specific genes, and the resulting generation of selected, pre-programmed cell types may represent an additional strategy to adapt to certain host environments . Finally, C . albicans produces genetically altered variants at a high rate . This microevolution ensures survival when the pathogen encounters new adverse conditions, as exemplified by the development of stable drug-resistant variants under the selection pressure caused by antimycotic therapy . Thus, rather than the possession of single dominant virulence factors, it is its remarkable versatility that makes C . albicans the most important fungal pathogen of humans.

J Behav Med, 2001 Jun, 24(3), 219 - 29
Differences in functional immune responses of high vs . low hardy healthy individuals; Dolbier CL et al.; An association between the personality trait of hardiness and immune responses was explored . Blood samples were collected from 21 healthy individuals under nonstressful conditions, who had either high or low levels of hardiness . Functional immune assays tested for natural killer (NK) cell activity and proliferation responses to Candida albicans (Candida), purified protein derivative from Mycobacterium tuberculosis (PPD), lipopolysaccharide (LPS), concanavalin A (Con A), and Staphylococcus enterotoxin A (Staph A) . Differences between high and low hardy groups on these immune responses were examined using Bonferroni adjusted independent t-tests . Results revealed significant differences between the groups for Candida, PPD, Con A, and Staph A . In all instances, the high hardy group had larger mean proliferative responses than the low hardy group . Implications of the study as well as avenues for future research are discussed.

J Leukoc Biol, 2001 Jul, 70(1), 149 - 54
Role of extracellular signal-regulated protein kinase cascade in macrophage killing of Candida albicans; Ibata-Ombetta S et al.; The pathogenic yeast Candida albicans and its derived molecules stimulate a wide range of macrophage secretory functions and may adapt to escape being killed by this phagocyte . In this study, phagocytosis of C . albicans and of the nonpathogenic yeast Saccharomyces cerevisiae was shown to be associated with phosphorylation of the mitogen-activated protein kinase (MAPK)/extracellularly regulated kinase (ERK) pathway in the absence of significant activation of either p38MAPK or stress-activated protein kinase/c-Jun N-terminal kinase . However, although 80% of endocytosed C . albicans survived after 1 h, 80% of S . cerevisiae cells were killed . Considerable quantitative differences were observed between the two species in the sequential phosphorylation of MAPK/ERK kinase (MEK), extracellularly regulated kinase-1, and 90-kDa-ribosomal S6 kinases . A lower level of activation of the pathway by C . albicans was associated with a species-specific overexpression of the MEK phosphatase MAPK phosphatase (MKP)-1 . Killing of both C . albicans and S . cerevisiae could be reduced using PD98059, which mimics MKP-1 and inhibits MEK phosphorylation, suggesting that specific MKP-1 activation by C . albicans could contribute to its ability to escape the yeast lytic potential of macrophages.

J Leukoc Biol, 2001 Jul, 70(1), 130 - 4
Mechanism of extracellular release of human neutrophil calprotectin complex; Voganatsi A et al.; Calprotectin is an abundant cytosolic protein complex of human neutrophils with in vitro extracellular antimicrobial activity . Studies suggest that calprotectin may be actively secreted from intact HL-60 cells and that it can be translocated to polymorphonuclear neutrophil (PMN) cell membranes . To examine whether calprotectin is secreted extracellularly, we incubated soluble and particulate stimuli, including live and heat-inactivated Candida albicans, with whole blood and measured calprotectin levels in the plasma . We compared the release of calprotectin to that of lactoferrin, a protein known to be secreted by PMNs . Extracellular lactoferrin was detected after incubation with any of the particulate stimuli . In contrast, a significant increase in extracellular calprotectin was found only after incubation with live C . albicans . Specifically, the increase in extracellular calprotectin correlated directly with a proportional decrease in PMN viability . Our results indicate that human PMN calprotectin is not secreted extracellularly except as a result of cell disruption or death.

Trends Microbiol, 2001 Jul, 9(7), 327 - 35
Virulence factors of Candida albicans; Calderone RA et al.; Candidiasis is a common infection of the skin, oral cavity and esophagus, gastrointestinal tract, vagina and vascular system of humans . Although most infections occur in patients who are immunocompromised or debilitated in some other way, the organism most often responsible for disease, Candida albicans, expresses several virulence factors that contribute to pathogenesis . These factors include host recognition biomolecules (adhesins), morphogenesis (the reversible transition between unicellular yeast cells and filamentous, growth forms), secreted aspartyl proteases and phospholipases . Additionally, 'phenotypic switching' is accompanied by changes in antigen expression, colony morphology and tissue affinities in C . albicans and several other Candida spp . Switching might provide cells with a flexibility that results in the adaptation of the organism to the hostile conditions imposed not only by the host but also by the physician treating the infection.

J Nat Prod, 2001 Feb, 64(2), 262 - 4
Three new peroxides from the sponge Plakinastrella species; Chen Y et al.; Two new five-membered-ring peroxide acids, plakinic acid F (3) and epiplakinic acid F (4), and a new peroxide-lactone, plakortolide F (5), were isolated from a sponge of the genus Plakinastrella collected from Felicite Island, Seychelles . The structures were elucidated through spectral analysis . The free acids 3 and 4 exhibit moderate antifungal activity against Candida albicans with minimum inhibitory concentrations of 25 micrograms/mL (SDB) and 3.1 micrograms/mL (RPMI) for 3, and 25 micrograms/mL (SDB) and 6.25 micrograms/mL (RPMI) for 4, respectively . Both also showed moderate in vitro inhibition of Aspergillus fumigatus with IC90's of 25 micrograms/mL.

J Nat Prod, 2001 Feb, 64(2), 154 - 8
Two novel cyclic peptides with antifungal activity from the cyanobacterium Tolypothrix byssoidea (EAWAG 195); Jaki B et al.; Two novel cyclic tridecapeptides, tolybyssidins A (1) and B (2), were isolated from the culture medium of mass cultured cyanobacterium Tolypothrix byssoidea (EAWAG 195) by means of bioguided isolation . The gross structures of these peptides were determined by 1D and 2D NMR experiments and tandem mass spectrometry . Both peptides contain the nonnatural amino acid dehydrohomoalanine (Dhha) as well as proteinogenic amino acids albeit with D- or L-configuration . The compounds exhibit moderate antifungal activity against the yeast Candida albicans.

Microbiology, 2001 Jul, 147(Pt 7), 1983 - 91
Secretion, interaction and assembly of two O-glycosylated cell wall antigens from Candida albicans; Pavia J et al.; The mechanisms of incorporation of two antigens have been determined using a monoclonal antibody (3A10) raised against the material released from the mycelial cell wall by zymolyase digestion and retained on a concanavalin A column . One of the hybridomas secreted an IgG that reacted with two bands in Western blots . Indirect immunofluorescence showed that the antigens were located on the surfaces of mycelial cells, but within the cell walls of yeasts . These antigens were detected in a membrane preparation, in the SDS-soluble material and in the material released by a 1,3-beta-glucanase and chitinase from the cell walls of yeast and mycelial cells . In the latter three samples, an additional high-molecular-mass, highly polydispersed band was also detected . Beta-elimination of each fraction resulted in the disappearance of all antigen bands, suggesting that they are highly O-glycosylated . In addition, the electrophoretic mobility of the high-molecular-mass, highly polydispersed bands increased after digestion with endoglycosidase H, indicating that they are also N-glycosylated . New antigen bands were released when remnants of the cell walls extracted with 1,3-beta-glucanase or chitinase were digested with chitinase or 1,3-beta-glucanase . These results are consistent with the notion that, after secretion, parts of the O-glycosylated antigen molecules are transferred to an N-glycosylated protein(s) . This molecular complex, as well as the remaining original 70 and 80 kDa antigen molecules, next bind to 1,3-beta-glucan or chitin, probably via 1,6-beta-glucan, and, in an additional step, to chitin or 1,3-beta-glucan . This process results in the final molecular product of each antigen, and their distribution in the cell walls.

Microbiology, 2001 Jul, 147(Pt 7), 1971 - 81
Different adhesins for type IV collagen on Candida albicans: identification of a lectin-like adhesin recognizing the 7S(IV) domain; Alonso R et al.; Adherence of the opportunistic pathogen Candida albicans to basement membrane (BM) proteins is considered a crucial step in the development of candidiasis . In this study the interactions of C . albicans yeast cells with the three main domains of type IV collagen, a major BM glycoprotein, were analysed . C . albicans adhered to the three immobilized domains by different mechanisms . Adhesion to the N-terminal cross-linking domain (7S) required the presence of divalent cations, whereas interaction with the central collagenous domain (CC) was cation-independent . Recognition of the C-terminal non-collagenous domain (NC1) was partially cation-dependent . Binding inhibition assays with the corresponding domains in soluble form showed that these interactions were specific . Both Ca(2+) and Mg(2+) promoted adhesion to the 7S domain and the interaction was completely abolished by EDTA . Treatment of the 7S domain, or its subunits, with N-glycosidase F reduced yeast binding by approximately 70% . Moreover, several sugars known to be part of the N-linked oligosaccharide chains of collagen IV inhibited adhesion to immobilized 7S; N-acetylglucosamine, L-fucose and methylmannoside caused a similar inhibition whereas N-acetyllactosamine was a more effective inhibitor . In contrast, glucose, galactose, lactose or heparan sulfate did not affect yeast binding . Combinations of the inhibitory sugars at suboptimal inhibition concentrations did not reduce C . albicans adhesion more than the individual sugars, pointing to a single lectin as responsible for the interaction . These results taken together show that C . albicans utilizes several adhesins for interacting with type IV collagen, and that at least one of them is a lectin which recognizes the 7S(IV) oligosaccharide residues as its receptor.

Microbiology, 2001 Jul, 147(Pt 7), 1961 - 70
Overexpression of a dominant-negative allele of YPT1 inhibits growth and aspartyl protease secretion in Candida albicans; Lee SA et al.; To investigate the pre-Golgi secretion pathway in the pathogenic yeast Candida albicans, we cloned the C . albicans homologue of the Saccharomyces cerevisiae protein secretion gene YPT1 . The C . albicans YPT1 ORF contained a 624 bp intronless ORF encoding a deduced protein of 207 aa and 2.3 kDa . This deduced protein was 77% identical to S . cerevisiae Ypt1 protein (Ypt1p) and it contained GTP-binding domains that are conserved among all known ras-like GTPases . Multicopy plasmids containing C . albicans YPT1 complemented the temperature-sensitive S . cerevisiae ypt1 (A136D) mutation . One chromosomal YPT1 allele in C . albicans CAI4 was readily disrupted by homologous gene targeting, but attempts to disrupt the second allele yielded no viable null mutants . Since this suggested that C . albicans YPT1 may be essential, a mutant ypt1 allele was constructed encoding the amino acid substitution analogous to the N121I substitution in a known trans-dominant inhibitor of S . cerevisiae Ypt1p . Next, a GAL1-regulated plasmid was used to express the mutant ypt1(N121I) allele in C . albicans CAI4 . Ten of 11 transformants tested grew normally in glucose and poorly in galactose, and plasmid curing restored growth to wild-type levels . When these transformants were incubated in galactose, secretion of aspartyl proteinase (Sap) was inhibited and membrane-bound secretory vesicles accumulated intracellularly . These results imply that C . albicans YPT1 is required for growth and protein secretion, and they confirm the feasibility of using inducible dominant-negative alleles to define the functions of essential genes in C . albicans.

Microbiology, 2001 Jul, 147(Pt 7), 1955 - 9
A diffusible analogue of N(3)-(4-methoxyfumaroyl)-L-2,3-diaminopropanoic acid with antifungal activity; Zgodka D et al.; N(3)-(4-Methoxyfumaroyl)-L-2,3-diaminopropanoic acid (FMDP), a specific and potent inactivator of glucosamine-6-phosphate (GlcN-6-P) synthase from Candida albicans, exhibits relatively poor anticandidal activity, with an MIC value amounting to 50 microg ml(-1) (200 microM) . Uptake of FMDP into C . albicans cells follows saturation kinetics and is sensitive to the action of metabolic inhibitors, thus indicating the active transport mechanism . However, the acetoxymethyl ester of FMDP penetrates the fungal cell membrane by free diffusion and is rapidly hydrolysed by C . albicans cytoplasmic enzymes to release the free FMDP . This mechanism gives rise to continuous accumulation of the enzyme inhibitor and results in higher antifungal activity of the FMDP ester (MIC=3.1 microg ml(-1), 10 microM) . These results show that the 'pro-drug' approach can be successfully applied for the enhancement of antifungal activity of glutamine analogues that inhibit GlcN-6-P synthase.

Transpl Infect Dis, 1999 Dec, 1(4), 262 - 72
Approaches to fungal diagnosis in transplantation; Patterson TF; The diagnosis of invasive fungal infection in patients undergoing solid organ or bone marrow transplantation remains a significant clinical challenge . Consideration of the epidemiology of these infections and host risk factors may be an important clue to a specific fungal diagnosis . Despite extensive investigation on methods such as serologic techniques to improve the rapid diagnosis of these infections, the diagnosis of invasive mycoses remains largely dependent on clinical presentation . For example, the signs and symptoms that result from angioinvasion of fungal organisms include pleuritic chest pain or hemoptysis . In a high-risk patient these findings can be important clues to invasive fungal infection . Cultures of opportunistic fungi in certain settings, such as Aspergillus in respiratory samples from immunosuppressed patients, may be associated with infection . Radiographic findings can also be useful to establish a diagnosis of infection . In patients with invasive aspergillosis as well as other angioinvasive moulds, chest CT scans may demonstrate lesions that are not visible on plain radiographs . Serodiagnosis of these infections remains largely investigational . Microbiological antifungal resistance has increasingly been reported, but in patients at high risk for serious fungal infection, including patients undergoing bone marrow and organ transplantation, antifungal resistance remains uncommon, particularly in Candida albicans . Higher doses of azoles should be used to treat patients with infections due to less susceptible yeasts and those with more serious infection . Prompt recognition of fungal infection combined with intensive antifungal therapy is needed for successful therapy.

J Hosp Infect, 2001 Jun, 48(2), 122 - 8
Decrease in Candida albicans strains with reduced susceptibility to fluconazole following changes in prescribing policies; Lopez J et al.; This study was undertaken to identify prescribing policies likely to favour or limit fluconazole resistance within a clinical department . Fluconazole exposure within the infectious diseases and clinical haematology units was investigated, and data were compared with in vitro susceptibility of Candida albicans isolates obtained in these units . Fluconazole utilization was determined by the number of fluconazole treatment-days per 100 hospitalization days (penetration index) . In the infectious diseases unit, separate evaluations for low-dose fluconazole (50 mg) prescribed as intermittent or prolonged treatment, and for higher-dosing schedules (fluconazole 200 mg) were made . Susceptibility of C . albicans isolates was surveyed in a broth microdilution assay by measuring the inhibitory concentration 50% (IC50) . The penetration index (PI) for fluconazole 50mg declined from 1992 to 1977 in infectious diseases (P= 0.0048) . In the meantime, total usage of fluconazole increased, due to increased prescribing of fluconazole 200 mg (P = 0.0724) . The IC50 of C . albicans isolates tested in infectious diseases decreased between 1994 and 1996 from 7.33 mg/ml to 1.64 mg/ml (P = 0.0075) . In clinical haematology, declines in C . albicans IC50 and fluconazole PI were not significant (P = 0.35 and P = 0.07, respectively) . These data suggest that prolonged or repeated exposure to low-dose fluconazole, rather than total cumulative use, was associated with fluconazole resistance in the infectious diseases unit . Moreover, restoration of a normal ecology was observed when low-dose prolonged or intermittent prescriptions were reduced .

Yeast, 2001 Jun 30, 18(9), 859 - 64
Cassettes for PCR-mediated construction of green, yellow, and cyan fluorescent protein fusions in Candida albicans; Gerami-Nejad M et al.; We have developed a set of plasmids containing fluorescent protein cassettes for use in PCR-mediated gene tagging in Candida albicans . We engineered YFP and CFP variants of the GFP sequence optimized for C . albicans codon usage . The fluorescent protein sequences, linked to C . albicans auxotrophic marker sequences, were amplified by PCR and transformed directly into yeast . Gene-specific sequence was incorporated into the PCR primers, such that the tag-cassette integrates by homologous recombination at the 3'-end of the gene of interest . This technique was used to tag Cdc3 and Tub1 with GFP, YFP and CFP, which were readily visualized by fluorescence microscopy and localized as expected . In addition, Tub1-YFP and Cdc3-CFP were visualized in the same cells . Thus, this technique directs one-step construction of multiple fluorescent protein fusions, facilitating the study of protein co-expression and co-localization in C . albicans cells in vivo .

Acta Biol Hung, 2001, 52(2-3), 289 - 98
Variation of isoenzyme and RAPD patterns in Candida albicans morphological mutants with altered colony ultrastructure; Pesti M et al.; Molecular typing methods were applied to characterize four stable morphological mutants {1} isolated from a UV-induced unstable mutant colony of Candida albicans . The wild-type strain (ATCC 64385), the intermediate unstable mutant and its four morphologically altered derivatives revealed the same electrophoretic karyotypes . Of the five isoenzymes tested (catalase, malate dehydrogenase, glutamate dehydrogenase, acid phosphatase and 3-glucosidase), glutamate dehydrogenase displayed a different enzyme pattern (with an extra band of lower mobility) in the morphological mutants . In contrast, the random amplification DNA polymorphism patterns of the mutant strains differed in all cases from that of the parental strain . Different primers revealed various degrees of DNA polymorphism; one of them (OPC-8) proved to be useful for differentiation between all examined strains . Differences in genetic alterations between spontaneous and induced mutants, and the applicability of different molecular markers to analyse the consequences of induced mutagenesis in C . albicans are discussed.

Appl Environ Microbiol, 2001 Jul, 67(7), 2982 - 92
Quorum sensing in the dimorphic fungus Candida albicans is mediated by farnesol; Hornby JM et al.; The inoculum size effect in the dimorphic fungus Candida albicans results from production of an extracellular quorum-sensing molecule (QSM) . This molecule prevents mycelial development in both a growth morphology assay and a differentiation assay using three chemically distinct triggers for germ tube formation (GTF): L-proline, N-acetylglucosamine, and serum (either pig or fetal bovine) . In all cases, the presence of QSM prevents the yeast-to-mycelium conversion, resulting in actively budding yeasts without influencing cellular growth rates . QSM exhibits general cross-reactivity within C . albicans in that supernatants from strain A72 are active on five other strains of C . albicans and vice versa . The QSM excreted by C . albicans is farnesol (C(15)H(26)O; molecular weight, 222.37) . QSM is extracellular, and is produced continuously during growth and over a temperature range from 23 to 43 degrees C, in amounts roughly proportional to the CFU/milliliter . Production is not dependent on the type of carbon source nor nitrogen source or on the chemical nature of the growth medium . Both commercial mixed isomer and (E,E)-farnesol exhibited QSM activity (the ability to prevent GTF) at a level sufficient to account for all the QSM activity present in C . albicans supernatants, i.e., 50% GTF at ca . 30 to 35 microM . Nerolidol was ca . two times less active than farnesol . Neither geraniol (C(10)), geranylgeraniol (C(20)), nor farnesyl pyrophosphate had any QSM activity.

J Dent Educ, 2001 May, 65(5), 463 - 79
The human genome, implications for oral health and diseases, and dental education; Slavkin HC; We are living in an extraordinary time in human history punctuated by the convergence of major scientific and technological progress in the physical, chemical, and biological ways of knowing . Equally extraordinary are the sparkling intellectual developments at the interface between fields of study . One major example of an emerging influence on the future of oral health education is at the interface between the human genome, information technology, and biotechnology with miniaturizations (nanotechnology), suggesting new oral health professional competencies for a new century . A great deal has recently been learned from human and non-human genomics . Genome databases are being "mined" to prompt hypothesis-driven "postgenomic" or functional genomic science in microbial models such as Candida albicans related to oral candidiasis and in human genomics related to biological processes found in craniofacial, oral, and dental diseases and disorders . This growing body of knowledge is already providing the gene content of many oral microbial and human genomes and the knowledge of genetic variants or polymorphisms related to disease, disease progression, and disease response to therapeutics (pharmacogenomics) . The knowledge base from human and non-human genomics, functional genomics, biotechnology, and associated information technologies is serving to revolutionize oral health promotion, risk assessment using biomarkers and disease prevention, diagnostics, treatments, and the full range of therapeutics for craniofacial, oral, and dental diseases and disorders . Education, training, and research opportunities are already transforming the curriculum and pedagogy for undergraduate science majors, predoctoral health professional programs, residency and specialty programs, and graduate programs within the health professions . In the words of Bob Dylan, "the times they are a-changing."

J Oral Rehabil, 2001 Jun, 28(6), 526 - 33
Candida albicans colonization on thermal cycled maxillofacial polymeric materials in vitro; Nikawa H et al.; In the present study, the colonization of a single isolate of Candida albicans on saliva-, serum-coated or protein free (uncoated), thermocycled (4-70 degrees C for 1 min, respectively; 0, 1000 and 10 000 times) 15 commercial maxillofacial materials were investigated, by adenosine triphosphate (ATP) analysis . In the case of control specimens (not thermocycled and uncoated), the fungal colonization appeared to depend upon the type of commercial products used . Thus, the lowest colonization was observed with additional silicone materials, soft acrylic liners with visible light curing, except for one product, whereas visible light curing liners comprising of single paste or single gel exhibited the highest colonization capacity, and cold cured acrylic liners exhibited the intermediate . However, the fungal colonization on the materials was significantly promoted both by thermal cycling (ANOVA; P < 0.01) and a layer of protein coating (saliva, P < 0.01; serum, P < 0.01) . When the inter-relation between the fungal colonization and the surface hydrophobicity of the materials were analysed, fungal colonization on 1000- and 10 000-thermocycled materials correlated well with the contact angles of the materials (Student's t-test, P < 0.01), being consistent with the thermodynamic theory . These results, taken together, suggest that the ageing of the materials and the biological fluids of the host promote yeast colonization on maxillofacial materials.

Retina, 2001, 21(3), 203 - 9
Analysis of predisposing clinical and laboratory findings for the development of endogenous fungal endophthalmitis . A retrospective 12-year study of 79 eyes of 46 patients; Tanaka M et al.; PURPOSE: To analyze the predisposing factors for the development of endogenous fungal endophthalmitis (EFE) for its early diagnosis and treatment . SUBJECTS AND METHODS: Seventy-nine eyes of 46 patients with EFE treated in the 12-year period between 1986 and 1998 were included . A retrospective analysis was conducted in respect to age, sex, underlying disease, visual acuity, findings in the anterior and posterior segments, fungal culture of surgical specimens, fever of unknown origin, neutrophils < or = 500/mL, Cand-tec > or = x 4, beta-D-glucan > or = 20 pg, and final visual acuity . RESULTS: The patients were 34 men (74%) and 12 women (26%) between 18 and 78 years of age (mean 57.2 years) . Thirty-three of the 46 patients (72%) also were diagnosed with cancer . Fungal infiltration limited to the retina (Stage I) was noted in 13%, budding in the vitreous cavity (Stage II) in 40%, vitreous opacity (Stage III) in 29%, and retinal detachment with Stage III (Stage IV) in 18% of 79 eyes with EFE . Forty patients (87%) were undergoing intravenous hyperalimentation (IVH) . The mean interval between the start of IVH and the onset of disease was 11 days . Vitreous surgery was performed in 26 eyes (33%) . Candida albicans was detected from surgical specimens in 38% . Fever of unknown origin was noted in 76%, neutrophils < or = 500/mL in 67%, Cand-tec > or = x 4 in 57%, and beta-D-glucan > or = 20 pg in 90% of subjects . CONCLUSION: In patients susceptible to opportunistic infection, beta-D-glucan > or = 20 pg (90%), IVH (87%), fever of unknown origin (76%), male sex (74%), the presence of cancer (72%), neutrophils < or = 500/mL (67%), and Cand-tec > or = x 4 (57%) were considered to be predisposing factors for the development of EFE.

Klin Monatsbl Augenheilkd, 2001 May, 218(5), 398 - 400
{Endogenous fungal endophthalmitis: results of antifungal treatment with and without vitrectomy}; Bagnoud M et al.; BACKGROUND: Fungal endophthalmitis represents a significant cause of ocular morbidity, affecting in the majority of cases patients in poor general conditions . The eye is typically involved by hematogen dissemination, and the germ induces a chorioretinitis associated with an important panuveitis . PATIENTS AND METHODS: Four patients were examined . They complained of a progressive reduction of vision, associated with photophobia . Ophthalmologic examination disclosed an important panuveitis . Investigations showed a fungal chorio-retinitis in all cases . Risk factors were intra-venous toxicomania, longterm parenteral nutrition and traumatism of the sinuses . RESULTS: Vitreous cultures were positive for Candida albicans (3 patients) and for Aspergillus fumigatus (1 patient) . Blood cultures were negative in the four cases . Three patients were treated with anti-fungal medication (fluconazole, itraconazole) associated with a vitrectomy by pars plana . One patient was treated by anti-fungal therapy only . Clinical evolution was satisfactory in all cases . Final vision was 10/10 in three cases and 5/10 in one . One patient developed a retinal detachment and an epiretinal membrane . Follow-up was 7 months (2-16 months) . CONCLUSION: The evolution of these four cases suggests that a rapid anti-fungal therapy associated with or without a vitrectomy represent a favourable therapeutic option when a fungal infection is suspected.

Mycoses, 2001 May, 44(3-4), 77 - 81
Incidence of Candida in psoriasis--a study on the fungal flora of psoriatic patients; Waldman A et al.; The presence of Candida albicans and other Candida species in saliva and faeces of 50 psoriatic patients compared with a control group of 50 healthy donors was examined quantitatively . The quantity of Candida in saliva and faeces of the psoriatics proved to be significantly higher than in the controls . Candida was detected in 78% of the saliva samples of the psoriatics but in only 50% of the controls, and in the faeces samples in 72% of the psoriatics, but in only 46% of the controls . Qualitative analysis revealed a predominance of Candida albicans (saliva, 77%; faeces, 64%) and Candida rugosa (saliva, 28%; faeces, 28%) . We did not find a correlation between the severity of the psoriasis according to the Psoriasis Area and Severity Index and the amount of Candida in the saliva or in the faeces . Our results reinforce the hypothesis that C . albicans is one of the triggers to both exacerbation and persistence of psoriasis . We propose that in psoriatics with a significant quantity of Candida in faeces, an antifungal treatment should be considered as an adjuvant treatment of psoriasis.

Arch Pharm (Weinheim), 2001 May, 334(5), 148 - 52
Synthesis and antimicrobial activity of some new 2-phenyl-N-substituted carboxamido-1H-benzimidazole derivatives; Goker H et al.; Some 1H-benzimidazole-carboxamide derivatives were prepared and their antimicrobial activities against Staphyloccus aureus, Escherichia coli and Candida albicans evaluated . Compounds 18, 22, and 25 exhibited the best activity against Candida albicans.

Nucleic Acids Res, 2001 Jun 15, 29(12), 2644 - 53
Folding of the group I intron ribozyme from the 26S rRNA gene of Candida albicans; Zhang Y et al.; Preincubation of the group I intron Ca.LSU from Candida albicans at 37 degrees C in the absence of divalent cations results in partial folding of this intron . This is indicated by increased resistance to T1 ribonuclease cleavage of many G residues in most local helices, including P4-P6, as well as the non-local helix P7, where the G binding site is located . These changes correlate with increased gel mobility and activation of catalysis by precursor RNA containing this intron after preincubation . The presence of divalent cations or spermidine during preincubation results in formation of the predicted helices, as indicated by protection of additional G residues . However, addition of these cations during preincubation of the precursor RNA alters its gel mobility and eliminates the preincubation activation of precursor RNA seen in the absence of cations . These results suggest that, in the presence of divalent cations or spermidine, Ca.LSU folds into a more ordered, stable but misfolded conformation that is less able to convert into the catalytically active form than the ribozyme preincubated without cations . These results indicate that, like the group I intron of Tetrahymena, multiple folding pathways exist for Ca.LSU . However, it appears that the role cations play in the multiple folding pathways leading to the catalytically active form may differ between folding of these two group I introns.

Eur J Med Res, 2001 May 29, 6(5), 193 - 200
Bromelain is an accelerator of phagocytosis, respiratory burst and Killing of Candida albicans by human granulocytes and monocytes; Brakebusch M et al.; OBJECTIVE: The aim of this study was to examine the influence of immuno modulating agents like bromelain and trypsin (e.g . Wobenzym on granulocyte and monocyte functions in healthy volunteers and patients with disorders of the humoral immuno system X-linked agammaglobulinaemia (XLA) and common variable immuno deficiency (CVID) and to find out whether the unspecific immunity could be improved by these enzymes . METHODS: In a whole-blood assay kinetics of phagocytosis, respiratory burst and killing (PBK) were measured in blood samples incubated with and without bromelain and trypsin (B/T) using Candida albicans as target organism . The time-reaction curves were analysed determining their gradient (T1) and their onset (T2) as well as the half effect time (HET) . RESULTS: Phagocytes from patients with XLA showed a significantly accelerated basal phagocytosis (reduction of HET by 24% p < 0.001) compared to healthy controls . After incubation with B/T (10 microg/ml each) speed of phagocytosis was nearly doubled (phagocyte activity p < 0.0001, Candida uptake p < 0.003), T2 of respiratory burst was reduced by 65 % (p < 0.0001) and killing was accelerated by 27% (p < 0.046) . However, the maximal activities of all kinetics were not altered . Incubation of phagocytes from healthy controls with B/T accelerated phagocytosis to a level comparable to that of untreated phagocytes from patients with XLA and also accelerated reactive oxygen species (ROS) production (reduction of HET by 28%, p < 0.012) . In contrast to phagocytes from patients with XLA, phagocytes of patients with CVID showed a similar stimulation by B/T like healthy controls . Further experiments with the single substances showed that bromelain was the active compound . CONCLUSION: Our data suggest, that bromelain possesses immuno stimulatory properties . Phagocytes of XLA patients appear to be particularly susceptible to this stimulation.

Genetics, 2001 Jun, 158(2), 919 - 24
A histone deacetylation inhibitor and mutant promote colony-type switching of the human pathogen Candida albicans; Klar AJ et al.; Most strains of Candida albicans undergo high frequency phenotypic switching . Strain WO-1 undergoes the white-opaque transition, which involves changes in colony and cellular morphology, gene expression, and virulence . We have hypothesized that the switch event involves heritable changes in chromatin structure . To test this hypothesis, we transiently exposed cells to the histone deacetylase inhibitor trichostatin-A (TSA) . Treatment promoted a dramatic increase in the frequency of switching from white to opaque, but not opaque to white . Targeted deletion of HDA1, which encodes a deacetylase sensitive to TSA, had the same selective effect . These results support the model that the acetylation of histones plays a selective role in regulating the switching process.

Int Arch Allergy Immunol, 2001, 125 Suppl 1, 48 - 50
IL-5 production by peripheral blood Th cells of adult asthma patients in response to Candida albicans allergen; Mori A et al.; Cytokines produced by T cells are involved in chronic asthma . Relevant antigens for nonatopic asthma are still mostly unknown . Peripheral blood mononuclear cells (PBMCs) were obtained from adult asthmatic donors and incubated with Candida albicans extract . Proliferation response and cytokine production were analyzed . IL-2 and IL-5 were detectable in the cultures incubated with C . albicans extract, whereas IL-4 was undetectable . PBMCs obtained from some nonatopic asthma patients produced IL-5 upon stimulation of C . albicans extract . C . albicans allergen may be involved in the airway eosinophilic inflammation through the induction of IL-5 production by Th cells .

Antimicrob Agents Chemother, 2001 Jul, 45(7), 2129 - 33
Prospective study of Candida species in patients at a comprehensive cancer center; Safdar A et al.; Since most nosocomial systemic yeast infections arise from the endogenous flora of the patient, we prospectively evaluated the species stratification and antifungal susceptibility profile of Candida spp . associated with heavy colonization and systemic infection in patients at Memorial Sloan-Kettering Cancer Center in New York . A total of 349 Candida isolates were obtained from 223 patients during the later half of 1998 . Cancer was the most common underlying disease, occurring in 91% of the patients, including 61.8% with organ and 23.7% with hematological malignancies; 4.4% of the patients had AIDS . Candida albicans was the predominant species (67.3%); among 114 non-albicans Candida spp., C . glabrata (45.6%) was the most frequent, followed by C . tropicalis (18.4%), C . parapsilosis (16.6%), and C . krusei (9.6%) . The overall resistance to triazole-based agents among all yeast isolates was 9.4 and 10.8% for fluconazole and itraconazole, respectively . A total of 5% of C . albicans strains were resistant to triazole antifungals, whereas 30.8 and 46.2% of C . glabrata strains were resistant to fluconazole (MIC > or = 64 microg/ml) and itraconazole (MIC > or = 1 microg/ml), respectively . A significant association was observed between prior treatment with triazole and isolation of fluconazole-resistant C . albicans (P = 0.005, OR 36), although this relationship was not seen in C . glabrata isolates (P = 0.4) . This study reinforces the importance of periodic, prospective surveillance of clinical fungal isolates to determine appropriate prophylactic, empiric, and preemptive antifungal therapy for the highly susceptible patient population.

Antimicrob Agents Chemother, 2001 Jul, 45(7), 2018 - 22
Influence of human serum on antifungal pharmacodynamics with Candida albicans; Zhanel GG et al.; Antifungal susceptibilities (NCCLS, approved standard M27-A, 1997) were determined for the reference strain ATCC 90028 and 21 clinical isolates of Candida albicans with varying levels of fluconazole susceptibility using RPMI 1640 (RPMI) and 80% fresh human serum-20% RPMI (serum) . Sixty-four percent (14 of 22) of the isolates tested demonstrated significant decreases (> or = 4-fold) in fluconazole MICs in the presence of serum, and the remaining eight isolates exhibited no change . Itraconazole and ketoconazole, two highly protein-bound antifungal agents, had MICs in serum that were increased or unchanged for 46% (10 of 22) and 41% (9 of 22) of the isolates, respectively . All 10 isolates tested against an investigational antifungal agent, LY303366, demonstrated significant increases in the MIC required in serum, while differences in amphotericin B MICs in the two media were not observed . Four of 10 isolates tested demonstrated fourfold higher flucytosine MICs in serum than in RPMI . Postantifungal effects (PAFEs) and 24-h kill curves were determined by standard methods for selected isolates . At the MIC, fluconazole, itraconazole, ketoconazole, flucytosine, and LY303366 kill curves and PAFEs in RPMI were similar to those in serum . Isolates of fluconazole-resistant C . albicans required lower MICs in serum than in RPMI, without relative increases in fungal killing or PAFEs . Isolates tested against amphotericin B demonstrated significantly reduced killing and shorter PAFEs in serum than in RPMI without observable changes in MIC . In conclusion, antifungal pharmacodynamics in RPMI did not consistently predict antifungal activity in serum for azoles and amphotericin B . Generally speaking, antifungal agents with high protein binding exhibited some form of reduced activity (MIC, killing, or PAFE) in the presence of serum compared to those with low protein binding.

Chemotherapy, 2001 Jul-Aug, 47(4), 279 - 91
Evaluation of polyene-azole antagonism in liquid cultures of Candida albicans using an automated turbidometric method; Samaranayake YH et al.; BACKGROUND: A computerized machine, SPECTRAmax 340, was used to evaluate the recently reported phenomenon of antagonism of the polyene amphotericin B (AMB) in Candida albicans pre-exposed to the triazole fluconazole (FLZ) . METHODS: We investigated growth inhibition by varying concentrations of AMB in seven isolates of C . albicans pre-exposed to FLZ (50 microg/ml) for 18 h . All isolates were obtained on sequential visits from human immunodeficiency virus-infected patients not treated with FLZ . RESULTS: Antagonism of AMB activity was observed in 5, 4, 2 and a single isolate for 0.5, 1, 2 and 3 microg/ml of the antifungal, respectively . In the majority of Candida isolates, antagonism was seen within a concentration range of 0.5-1.0 microg/ml AMB; 1 Candida strain (HK1-Sa) was resistant to 3 microg/ml AMB . Higher concentrations of AMB (>3 microg/ml) killed both the controls and FLZ-pre-exposed Candida cells . No significant differences were observed between the periods of antagonism observed for any of the sequential isolates or for any of the AMB concentrations or in the maxima of the growth curves obtained for all Candida isolates . CONCLUSION: We conclude that the SPECTRAmax system is a useful tool to evaluate in vitro pharmacodynamic interactions between antifungal regimens within a fluid culture system, and provides information that cannot be obtained using traditional plate assay systems.

Chemotherapy, 2001 Jul-Aug, 47(4), 261 - 5
In vitro effect of amikacin, imipenem, cefodizime, IFNalpha-2a alone and combinations of antibiotics with IFNalpha-2a on polymorphonuclear leukocyte function in chronic hepatitis patients; Adalati R et al.; The in vitro effect of amikacin (8 microg/ml), imipenem (30 microg/ml), cefodizime (10 microg/ml), interferon alpha-2a (IFNalpha-2a) (10 IU/ml) and antibiotic combinations with IFNalpha-2a on polymorphonuclear leukocyte (PMN) functions (phagocytosis and intracellular killing of Candida albicans blastospores) was investigated in chronic hepatitis B patients . Phagocytosis and candidacidal activity was not affected after pretreatment of PMNs with amikacin and imipenem (p > 0.05) . Phagocytic activity was enhanced after pretreatment of PMNs with cefodizime and IFNalpha-2a compared with that of control PMNs (p < 0.05), but candidacidal activity was not affected by the same drugs (p > 0.05) . Phagocytic and intracellular activity of PMNs were not affected by combinations of IFNalpha-2a and antibiotics (p > 0.05).

Phytother Res, 2001 Jun, 15(4), 337 - 43
Biological activity of kiwifruit peel extracts; Motohashi N et al.; Various bioactive substances in kiwifruit extracts were fractionated by organic solvent extractions, followed by silica gel and ODS chromatographies . Both cytotoxic activity and multi-drug resistance reversal activity were found in the less polar fractions . Cytotoxic activity was not always parallel the radical intensity . Antibacterial activity was distributed into various fractions and all fractions were inactive against Candida albicans and H . pylori . Only 70% methanol extracts showed anti-human immunodeficiency virus activity, and produced a broad ESR signal under alkaline conditions, in a fashion similar to lignin . These fractions also effectively scavenged O(2)(-) produced by the xanthine-xanthine oxidase reaction, suggesting a bimodal (pro-oxidant and antioxidant) action . These data suggest a medicinal efficacy of kiwifruit peel extracts .

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2001 Jun, 91(6), 659 - 62
Candida dubliniensis in radiation-induced oropharyngeal candidiasis; Redding SW et al.; Candida dubliniensis is a recently described species that has been shown to cause oropharyngeal candidiasis in patients with HIV . We present a detailed evaluation of a patient undergoing head and neck radiation for oral cancer who developed oropharyngeal candidiasis from a mixed infection of C dubliniensis and Candida albicans . To our knowledge, this is the first described case of C dubliniensis contributing to oropharyngeal candidiasis in this patient population.

J Biol Chem, 2001 Aug 17, 276(33), 31402 - 7 Epub 2001 Jun 11.
Identification of a putative sordarin binding site in Candida albicans elongation factor 2 by photoaffinity labeling; Dominguez JM et al.; Candida albicans EF-2 binds sordarin to a single class of binding sites with K(d) = 1.26 microm . Equimolar mixtures of EF-2 and ribosomes, in the presence of a non-hydrolyzable GTP analog, reveal two classes of high affinity sordarin binding sites with K(d) = 0.7 and 41.5 nm, probably due to the existence of two ribosome populations . Photoaffinity labeling of C . albicans EF-2 in the absence of ribosomes has been performed with {(14)C}GM258383, a photoactivatable sordarin derivative . Labeling is saturable and can be considered specific, because it can be prevented with another sordarin analog . The fragment Gln(224)-Lys(232) has been identified as the modified peptide within the EF-2 sequence, Lys(228) being the residue to which the photoprobe was linked . This fragment is included within the G"-subdomain of EF-2 . These results are discussed in the light of the high sordarin specificity toward fungal systems.

Infect Immun, 2001 Jul, 69(7), 4695 - 7
Hyphae and yeasts of Candida albicans differentially regulate interleukin-12 production by human blood monocytes: inhibitory role of C . albicans germination; Liu L et al.; The role of Candida albicans yeast-to-hyphae transition in interleukin-12 (IL-12) production by monocytes was investigated . Germinating C . albicans not only failed to induce IL-12 p70 but also suppressed IL-12 production induced by heat-killed C . albicans . Comparison of the abilities of germinating C . albicans and agerminating mutants to inhibit IL-12 production showed that germination of C . albicans plays a critical role in the inhibition of IL-12 production.

Infect Immun, 2001 Jul, 69(7), 4536 - 44
Traversal of Candida albicans across human blood-brain barrier in vitro; Jong AY et al.; Candida albicans is an opportunistic pathogen, which primarily affects neonates and immunocompromised individuals . The pathogen can invade the central nervous system, resulting in meningitis . At present, the pathogenesis of C . albicans meningitis is unclear . We used an in vitro model of the human blood-brain barrier to investigate the interaction(s) of C . albicans with human brain microvascular endothelial cells (BMEC) . Binding of C . albicans to human BMEC was time and inoculum dependent . Invasion of C . albicans into human BMEC was demonstrated by using an enzyme-linked immunosorbent assay based on fluorescent staining of C . albicans with calcoflour . In contrast, avirulent Candida mutant strains and nonpathogenic yeast Saccharomyces cerevisiae were not able to bind and invade human BMEC . Morphological studies revealed that on association with human BMEC, C . albicans formed germ tubes and was able to bud intracellularly . Transmission electron microscopy showed various stages of C . albicans interactions with human BMEC, e.g., pseudopod-like structures on human BMEC membrane and intracellular vacuole-like structures retaining C . albicans . Of interest, C . albicans was able to bud and develop pseudohyphae inside human BMEC without apparent morphological changes of the host cells . In addition, C . albicans penetrates through human BMEC monolayers without a detectable change in transendothelial electrical resistance and inulin permeability . This is the first demonstration that C . albicans is able to adhere, invade, and transcytose across human BMEC without affecting monolayer integrity . A complete understanding of the interaction(s) of C . albicans with human BMEC should contribute to the understanding of the pathogenic mechanism(s) of C . albicans meningitis.

Biochem Biophys Res Commun, 2001 Mar 30, 282(2), 570 - 4
Fungicidal effect of antimicrobial peptide, PMAP-23, isolated from porcine myeloid against Candida albicans; Lee DG et al.; The antifungal activity and mechanism of a 23-mer peptide, PMAP-23, derived from pig myeloid was investigated . PMAP-23 displayed strong antifungal activity against yeast and mold . To investigate the antifungal mechanism of PMAP-23, fluorescence activated flow cytometry and confocal laser scanning microscopy were performed . Candida albicans treated with PMAP-23 showed higher fluorescence intensity by propidium iodide(PI) staining, which was similar to that of Melittin than untreated cells . Confocal microscopy showed that the peptide was located in the plasma membrane . The action of peptides against fungal cell membranes was examined by treating prepared protoplasts of C . albicans with the peptide and lipid vesicle titration test . The result showed that the peptide prevented the regeneration of fungal cell walls and induced release of the fluorescent dye trapped in the artificial membrane vesicles, indicating that the peptide exerts its antifungal activity by acting on the plasma lipid membrane .

Mycoses, 2001, 44(1-2), 29 - 35
Adhesion of Candida parapsilosis to epithelial and acrylic surfaces correlates with cell surface hydrophobicity; Panagoda GJ et al.; We investigated in vitro adherence of 24 isolates of Candida parapsilosis and 12 isolates of Candida albicans with regard to their relative cell-surface hydrophobicity (CSH), adherence to human buccal epithelial cells (BEC) and acrylic surfaces . There was no significant interspecies difference in the relative adherence of C . parapsilosis and C . albicans isolates to BEC, although the former demonstrated a tendency for increased adherence . However, a significant intra-species variation in adherence among isolates of C . parapsilosis (P < 0.0001) to BEC, but not of C . albicans was noted . The superficial isolates of C . parapsilosis demonstrated a higher avidity (33%) to BEC than the systemic isolates . On regression analysis a significant positive correlation between C . parapsilosis adherence to BEC and denture acrylic surfaces was noted (r = 0.45, P = 0.02) . Similarly, buccal cell adherence correlated strongly with CSH of C . parapsilosis (r = 0.63, P = 0.0008) . These results shed further light on the intimate relationship between adherence and CSH in candidal colonization and imply that both C . parapsilosis and C . albicans are equally potent in colonizing mucosal surfaces with respect to the attributes investigated.

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