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% (C