Infect Immun, 2003 Oct, 71(10), 6101 - 3 Evaluation of the roles of four Candida albicans genes in virulence by using gene disruption strains that express URA3 from the native locus; Cheng S et al.; Reintroducing URA3 to its native locus in Candida albicans not5, not3, bur2, and kel1 disruption mutants enabled us to directly compare strains with control strain CAI-12 . We showed that URA3 position affected orotidine 5'-monophosphate decarboxylase activity, hyphal morphogenesis, adherence, and mortality in murine disseminated candidiasis . After URA3 was reintroduced to its native locus, only NOT5 could be conclusively ascribed a role in virulence. Infect Immun, 2003 Oct, 71(10), 5690 - 9 Deregulated production of protective cytokines in response to Candida albicans infection in patients with chronic mucocutaneous candidiasis; Lilic D et al.; Patients with chronic mucocutaneous candidiasis (CMC) are selectively unable to clear the yeast Candida, which results in persistent debilitating infections affecting the skin, nails, and mucous membranes . The underlying defect is unknown . Recent animal studies highlighted the importance of type 1 cytokines in protection against Candida, and previous work suggested that CMC patients may exhibit altered cytokine production in response to CANDIDA: Based on these findings, in this study we investigated cytokine production in CMC patients by assessing a range of inflammatory, anti-inflammatory, type 1, and type 2 cytokines (interleukin-2 {IL-2}, IL-4, IL-5, IL-6, IL-10, IL-12, gamma interferon {IFN-gamma}, tumor necrosis factor alpha {TNF-alpha}) in whole-blood cultures in response to five different fractions of Candida albicans (carbohydrate, purified mannan, and protein-rich fractions, etc.), as well as non-Candida antigens . Our results demonstrate that cytokine production is deregulated in a Candida-specific way for some cytokines (IL-2, IL-10), is deregulated more generally for other cytokines (IL-12, IL-6, IFN-gamma), and is not markedly altered for still other cytokines (TNF-alpha, IL-4, IL-5) . The most notable finding in CMC patients was the markedly impaired production of IL-12 in parallel with dramatically increased levels of IL-6 and IL-10 that occurred selectively in response to CANDIDA: These results suggest that patients with CMC have impaired production of type 1-inducing cytokines (possibly a macrophage or dendritic cell defect?), which could result in an inability to mount protective cell-mediated responses and a failure to clear CANDIDA: Continued tissue damage and inflammation may trigger production of high levels of inhibitory cytokines, such as the IL-10 production seen in our study, which would further reduce production of type 1-inducing cytokines in a positive feedback loop leading to persistent infection. J Microbiol Methods, 2003 Oct, 55(1), 321 - 3 Separation of the filamentous and cellular yeast forms of C . albicans following serum induction; Wansley DL et al.; Following serum induction of filamentous structures in Candida albicans, the budding and filamentous forms of the organism are separated and the total protein in each form is determined . The method is useful for testing potential chemotherapeutics aimed at interference with the formation of the pathogenic, filamentous form of C . albicans. Immunity, 2003 Sep, 19(3), 329 - 39 PPARgamma promotes mannose receptor gene expression in murine macrophages and contributes to the induction of this receptor by IL-13; Coste A et al.; Macrophage mannose receptor (MMR) is an important component of the innate immune system implicated in host defense against microbial infections such as candidiasis and in antigen presentation . We demonstrate here that the MMR expression is induced in mouse peritoneal macrophages following exposure to PPARgamma ligands or to interleukine-13 (IL-13) via a PPARgamma signaling pathway . Ligand activation of the PPARgamma in macrophages promotes uptake, killing of Candida albicans, and reactive oxygen intermediates production triggered by the yeasts through MMR overexpression . We also show that MMR induction by IL-13 via PPARgamma is dependent on phopholipase A2 activation and that IL-13 induces 15d-PGJ2 production and nuclear localization . These results reveal a novel signaling pathway controlling the MMR surface expression and suggest that endogenous PPARgamma ligand produced by phospholipase A2 activation may be an important regulator of MMR expression by IL-13. Intensive Care Med, 2003 Dec, 29(12), 2162 - 9 Epub 2003 Sep 10. Candidemia in critically ill patients: difference of outcome between medical and surgical patients; Charles PE et al.; OBJECTIVE: Candidemia is increasingly encountered in critically ill patients with a high fatality rate . The available data in the critically ill suggest that patients with prior surgery are at a higher risk than others . However, little is known about candidemia in medical settings . The main goal of this study was to compare features of candidemia in critically ill medical and surgical patients . DESIGN: Ten-year retrospective cohort study (1990-2000) . SETTING: Medical and surgical intensive care units (ICUs) of a teaching hospital . PATIENTS: Fifty-one patients with at least one positive blood culture for Candida species . MAIN RESULTS: Risk factors were retrieved in all of the patients: central venous catheter (92.1%), mechanical ventilation (72.5%), prior bacterial infection (70.6%), high fungal colonization index (45.6%) . Candida albicans accounts for 55% of all candidemia . The overall mortality was 60.8% (85% and 45.2% in medical and surgical patients, respectively) . Independent factors associated with survival were prior surgery (hazard ratio {HR} =0.25; 0.09-0.67 95% confidence interval {CI}, p<0.05), antifungal treatment (HR =0.11; 0.04-0.30 95% CI, p<0.05) and absence of neutropenia (HR =0.10; 0.02-0.45 95% CI, p<0.05) . Steroids, neutropenia and high density of fungal colonization were more frequently found among medical patients compared to surgical ones . CONCLUSIONS: Candidemia occurrence is associated with a high mortality rate among critically ill patients . Differences in underlying conditions could account for the poorer outcome of the medical patients . Screening for fungal colonization could allow identification of such high-risk patients and, in turn, improve outcome. Phytochemistry, 2003 Oct, 64(3), 743 - 52 Essential oil analysis and antimicrobial activity of eight Stachys species from Greece; Skaltsa HD et al.; The volatile composition of eight Stachys species has been studied . The investigated taxa were St . alopecuros (L.) Bentham., St . scardica (Griseb.) Hayek, St . cretica L . ssp . cretica, St . germanica L . ssp . heldreichii (Boiss.) Hayek, St . recta L., St . spinulosa L., St . euboica Rech . and St . menthifolia Vis., growing wild in Greece . The essential oils were obtained by hydrodistillation in a modified Clevenger-type apparatus, and their analyses were performed by GC and GC-MS . Identification of the substances was made by comparison of mass spectra and retention indices with literature records . Sesquiterpene hydrocarbons were shown to be the main group of constituents of all taxa . Furthermore, the obtained essential oils were tested against the following six bacteria: Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 35210), Bacillus subtilis (ATCC 10907), Bacillus cereus (clinical isolates), Micrococcus flavus (ATCC 10240), Staphylococcus epidermidis (ATCC 2228), as well as against the following five fungi: Aspergillus niger (ATCC 6275), Penicillium ochrochloron (ATCC 9112), Epidermophyton floccosum (clinical isolates), Candida albicans (clinical isolates) and Trichophyton mentagrophytes (clinical isolates) . The tested essential oils showed better activity against bacterial species than against fungi . Pseudomonas aeruginosa was the most resistant strain, as none of the essential oils was active against this strain . The essential oil of St . scardica has been proven most active against both bacteria and fungi. Int J Antimicrob Agents, 2003 Sep, 22(3), 291 - 300 Reversal of antifungal resistance mediated by ABC efflux pumps from Candida albicans functionally expressed in yeast; Schuetzer-Muehlbauer M et al.; The enhanced efflux of antifungal drugs through ATP-binding cassette (ABC) transporters constitutes a major cause of clinical multidrug resistance (MDR) . The inhibition of drug efflux pumps by specific compounds is considered to be a feasible strategy to overcome clinical antifungal resistance . Therefore, several blockers of mammalian and yeast ABC drug pumps, including FK506, propafenones, as well as the antifungal drug terbinafine were tested for their capacity to reverse CDR-mediated azole resistance in bakers yeast and in clinical isolates of Candida albicans . We have functionally expressed the C . albicans Cdr1p and Cdr2p transporters in hypersensitive Saccharomyces cerevisiae recipient strains lacking several endogenous ABC pumps . Cdr1p and Cdr2p were functional in yeast, as they conferred pronounced drug resistance to known antifungal drugs, including azoles and terbinafine . We employ two functional assays to demonstrate that ABC pump inhibitors reverse CDR-mediated antifungal resistance, thereby restoring drug susceptibility of yeast cells and resistant clinical isolates . Our results suggest that reversal of antifungal resistance can be achieved through ABC pump-dependent and independent mechanisms. FEMS Immunol Med Microbiol, 2003 Sep 22, 38(2), 173 - 80 Heterogeneity of metallo and serine extracellular proteinases in oral clinical isolates of Candida albicans in HIV-positive and healthy children from Rio de Janeiro, Brazil; de Brito Costa EM et al.; Candida yeasts frequently cause life-threatening systemic infections in immunocompromised hosts . In the present study, gelatin-SDS-PAGE analysis was used to characterize extracellular proteinases in 44 oral clinical isolates of Candida albicans from HIV-positive (29/50) and healthy children (15/50) . Our survey indicates that these oral clinical isolates of C . albicans have complex extracellular proteolytic activity profiles, which illustrates the heterogeneity of this species . We showed four distinct proteolytic patterns composed of distinct serine (30-58 kDa) and metalloproteinase (64-95 kDa) activities, based on the inhibition profile with phenylmethylsulfonyl fluoride and 1,10-phenanthroline, respectively . This is the first report on secreted serine and metalloproteinases present in the culture supernatant fluids of C . albicans; however, we did not observe a significant correlation between proteolytic profile expressed by the C . albicans isolates from HIV-positive children and CD4(+) T cell count and plasma viral load. FEMS Microbiol Lett, 2003 Sep 12, 226(1), 159 - 67 Characterization of a Candida albicans gene encoding a putative transcriptional factor required for cell wall integrity; Moreno I et al.; After screening a Candida albicans genome database the product of an open reading frame (ORF) (CA2880) with 49% homology to the product of Saccharomyces cerevisiae YPL133c, a putative transcriptional factor, was identified . The disruption of the C . albicans gene leads to a major sensitivity to calcofluor white and Congo red, a minor sensitivity to sodium dodecyl sulfate, a major resistance to zymolyase, and an alteration of the chemical composition of the cell wall . For these reasons we called it CaCWT1 (for C . albicans cell wall transcription factor) . CaCwt1p contains a putative Zn(II) Cys(6) DNA binding domain characteristic of some transcriptional factors and a PAS domain . The CaCWT1 gene is more expressed in stationary phase cells than in cells growing exponentially . To our knowledge, this is the first Zn(II) Cys(6) transcriptional factor-encoding gene implicated in the cell wall architecture. Bioorg Med Chem, 2003 Oct 1, 11(20), 4463 - 78 Synthesis and biological activities of benzofuran antifungal agents targeting fungal N-myristoyltransferase; Masubuchi M et al.; The C-4 side chain modification of lead compound 1 has resulted in the identification of a potent and selective Candida albicans N-myristoyltransferase (CaNmt) inhibitor RO-09-4609, which exhibits antifungal activity against C . albicans in vitro . Further modification of its C-2 substituent has led to the discovery of RO-09-4879, which exhibits antifungal activity in vivo . The drug design is based on X-ray crystal analysis of a CaNmt complex with benzofuran derivative 4a . The optimization incorporates various biological investigations including a quasi in vivo assay and pharmacokinetic study . The computer aided drug design, synthesis, structure-activity relationships, and biological properties of RO-09-4879 are described in detail. Southeast Asian J Trop Med Public Health, 2002, 33 Suppl 3, 152 - 4 Antifungal activity and local toxicity study of Alangium salviifolium subsp hexapetalum; Wuthi-udomlert M et al.; Alangium salviifolium subsp hexapetalum is a medicinal plant which has been traditionally used for tonic and treatment of hemorrhoid . This plant showed promising antimicrobial activity in our preliminary experiments, this study was, therefore, conducted to investigate its inhibitory effect against dermatomycotic organisms and its toxicity . The lyophilized powder extract (4.59%) of pulverized wood was tested for its inhibitory effect by agar disc diffusion test . The extract gave inhibitory zone diameters of 25.23 and 14.78 mm against 26 and 14 isolates of dermatophytes and Candida albicans, respectively . Ketoconazole, used as a reference antifungal agent, had inhibitory zone diameters of 33.15 and 27.93 mm against dermatophytes and C . albicans, respectively . There was no significant difference between the extract and ketoconazole in their inhibition against dermatophytes (p > 0.01), but their difference was significant against C . albicans (p < 0.01) . Using Buehler' s method, different amounts of extract (3, 6, and 9 mg/inch2 gauze pad) were tested in five male New Zealand white rabbits . All tested amounts of extract did not induce dermatitis among those rabbits within 1 week . The results demonstrated the inhibitory effect of Alangium salviifolium subsp hexapetalum against fungi without any local toxicity; a tendency to further develop a herbal preparation for the treatment of some dermatomycotic infections. Emerg Infect Dis, 2003 Aug, 9(8), 985 - 90 Candidemia in Finland, 1995-1999; Poikonen E et al.; We analyzed laboratory-based surveillance candidemia data from the National Infectious Disease Register in Finland and reviewed cases of candidemia from one tertiary-care hospital from 1995 to 1999 . A total of 479 candidemia cases were reported to the Register . The annual incidence rose from 1.7 per 100,000 population in 1995 to 2.2 in 1999 . Species other than Candida albicans accounted for 30% of cases without change in the proportion . A total of 79 cases of candidemia were identified at the hospital; the rate varied from 0.03 to 0.05 per 1,000 patient-days by year . Predisposing factors included indwelling catheters (81%), gastrointestinal surgery (27%), hematologic malignancy (25%), other types of surgery (21%), and solid malignancies (20%) . Crude 7-day and 30-day case-fatality ratios were 15% and 35%, respectively . The rate of candidemia increased in Finland but is still substantially lower than in the United States . No shift to non-C . albicans species could be detected. Pharmazie, 2003 Aug, 58(8), 527 - 30 Synthesis of some new benzothiazole derivatives as potential antimicrobial and antiparasitic agents; Mahran MA et al.; Several thiazolidinonyl benzothiazoles 8a-b and thiazolinylbenzothiazoles 9a-j were synthesized by the reaction of 2-(N-substituted thiocarbamoyl hydrazino) benzothiazoles 7a-d with chloroacetic acid or phenacyl bromide respectively . The intermediate compounds 7a-d were prepared in a good yield by the reaction of 2-hydrazinobenzothiazole (6) with phenylisothiocyanates . Synthesis of hydrazones 10a-c were performed by the reaction of 6 with the corresponding aldehydes . Trials to cyclize the obtained hydrazones 10a-c into the corresponding triazolo derivatives 11a-c were unsuccessful . Addition of 4-morphylino carbonyl chloride to compound 6 yielded the corresponding 2-acid hydrazide derivative 12 . Some of the prepared compounds were screened for their anti-parasitic activity . Most of them showed reasonable antinematodal or schistosomicidal activity . In addition, antimicrobial screening of all of the prepared new compounds was performed against Staphylococcus aeurus ATCC 6538, Escherichia coli ATCC 8735 and Candida albicans ATCC 10321 but non of them was active. Microbiol Mol Biol Rev, 2003 Sep, 67(3), 400 - 28, table of contents Candida albicans secreted aspartyl proteinases in virulence and pathogenesis; Naglik JR et al.; Candida albicans is the most common fungal pathogen of humans and has developed an extensive repertoire of putative virulence mechanisms that allows successful colonization and infection of the host under suitable predisposing conditions . Extracellular proteolytic activity plays a central role in Candida pathogenicity and is produced by a family of 10 secreted aspartyl proteinases (Sap proteins) . Although the consequences of proteinase secretion during human infections is not precisely known, in vitro, animal, and human studies have implicated the proteinases in C . albicans virulence in one of the following seven ways: (i) correlation between Sap production in vitro and Candida virulence, (ii) degradation of human proteins and structural analysis in determining Sap substrate specificity, (iii) association of Sap production with other virulence processes of C . albicans, (iv) Sap protein production and Sap immune responses in animal and human infections, (v) SAP gene expression during Candida infections, (vi) modulation of C . albicans virulence by aspartyl proteinase inhibitors, and (vii) the use of SAP-disrupted mutants to analyze C . albicans virulence . Sap proteins fulfill a number of specialized functions during the infective process, which include the simple role of digesting molecules for nutrient acquisition, digesting or distorting host cell membranes to facilitate adhesion and tissue invasion, and digesting cells and molecules of the host immune system to avoid or resist antimicrobial attack by the host . We have critically discussed the data relevant to each of these seven criteria, with specific emphasis on how this proteinase family could contribute to Candida virulence and pathogenesis. Med Mycol, 2003 Apr, 41(2), 167 - 70 Effect of ritonavir and saquinavir on Candida albicans growth rate and in vitro activity of aspartyl proteinases; Blanco MT et al.; An in vitro study to evaluate the antifungal effect and activity of aspartyl proteinases of the HIV-proteinase inhibitors ritonavir and saquinavir was conducted . Ritonavir diminished the growth rate of Candida albicans as well as the activity of its secreted aspartyl proteinases (Saps) in a nitrogen-limited medium, yeast carbon base and bovine serum albumin (YCB-BSA) . This inhibition occurred in a dose-dependent fashion; with 8 mg l(-1) of ritonavir a partial growth inhibition (44%) was produced . The growth rate of C . albicans in medium with saquinavir was similar to that seen in the control, and Sap activity was inhibited only at high concentrations . In conventional medium (RPMI-1640), which does not induce the production of yeast proteases, no inhibitory effect was detected with either HIV-protease inhibitor. Med Mycol, 2003 Apr, 41(2), 143 - 7 Genetic susceptibility to vaginal candidiasis; Calderon L et al.; To enable future studies on host resistance factors and therapy, inbred and outbred mouse strains were tested for susceptibility to vaginal candidiasis . Groups of mice were given 0.5 mg estradiol 3 days before and 4 days after intravaginal challenge with a suspension of Candida albicans . On day 1 after challenge, a swab was used to quantitate infection in all groups and to assure equivalent infection levels . On day 6, this was repeated and the experiment was terminated . BALB/c, the reference strain in repeated experiments, was susceptible, showing persistent infection with levels of cfu at day 6 falling within a range between a twofold decrease and a fourfold increase in relation to day 1 levels . CD-1 outbred mice were markedly resistant, with day 6 cfu levels showing a 74- to 87-fold decrease with respect to day 1 levels, whereas other outbred strains (CF-1, SW, ICR) were susceptible . A BALB/c substrain (ByJ) was also susceptible . With exception of CBA/J, which showed modest resistance, all inbred strains were similarly susceptible, including DBA/2, AKR/J, C3H/HeN, A/J and C57BL/6 . The differences between CD-1 and BALB/c mice were also seen with a second C . albicans isolate . Our results show susceptibility to vaginal candidiasis is independent of the major histocompatibility locus H2 haplotype and any effect ascribable to use of particular commercial mouse suppliers . Differences among mouse strains in susceptibility to C . albicans, as seen in previous studies involving nonvaginal challenge routes, are not reflected in this vaginal candidiasis model; in general, such resistance patterns appear specific to the route of challenge administration . The resistance seen in mouse strain CD-1 is of particular interest in that CD-1 is known to be resistant to endocrine disruption by estrogen . Our results suggest this estrogen insensitivity may have broad-ranging effects on processes other than gametogenesis, including vaginal susceptibility to candidiasis. Med Mycol, 2003 Apr, 41(2), 97 - 109 Local immune responsiveness following intravaginal challenge with Candida antigen in adult women at different stages of the menstrual cycle; Fidel PL Jr et al.; Recurrent vulvovaginal candidiasis (RVVC) is a significant problem in women of childbearing age and is most often caused by Candida albicans that asymptomatically colonizes mucosal tissues . Although some form of local immune dysfunction is postulated to precipitate bouts of RVVC, the normal protective vaginal host response to C . albicans is poorly understood . In an effort to stimulate the natural adaptive response to yeast in healthy women without a history of VVC, commercial Candida skin test antigen was introduced intravaginally and changes in cytokines/immunomodulators were monitored in vaginal lavage fluid pre- and post-antigen challenge . In an earlier pilot study using small numbers of women without controlling for stages of the menstrual cycle, we reported elevated cytokines in vaginal secretions of antigen challenged women . The present study, however, that employed a similar design in a large number of women during each stage of the menstrual cycle showed no evidence of local immune stimulation, including changes in Th and proinflammatory cytokines, IgE, histamine, and prostaglandin, despite a natural modulation of vaginal cytokines over the course of the menstrual cycle . Taken together, these results suggest that either some form of vaginal immunoregulation/tolerance is evident in response to yeast or that more advanced clinical designs are required to detect the normal protective vaginal response to C . albicans. Med Mycol, 2003 Aug, 41(4), 331 - 8 Infection of chick chorioallantoic membrane (CAM) as a model for invasive hyphal growth and pathogenesis of Candida albicans; Gow NA et al.; We report the development of a simple model for assessing the ability of the fungal pathogen Candida albicans to invade the chorioallantoic membrane (CAM) of fertilized hens' eggs . Wild-type and mutant strains of C . albicans were inoculated onto CAM surfaces either as a liquid suspension or on a sterile filter disc . Invasion of the membrane led to death of the embryo due to damage of the CAM, which could be examined histologically to show cell distribution and morphology, and by RT-PCR for assessment of patterns of fungal gene expression in vivo . Prophylactic or co-administration of fluconazole with the inoculum protected the embryo from infection . Secretory aspartyl protease (Sap) mutant strains with reported attenuation of virulence were virulent in the CAM model . However, a C . albicans strain with mutations in two transcription factors Efg1 and Cph1 was unable to form hyphae on the CAM or to penetrate it . The chick CAM, therefore, represents an experimentally tractable and inexpensive alternative to rodent or tissue culture-based invasion models, and can be used to investigate fungal pathogenesis and the genetic regulation of infection and membrane penetration of C . albicans. Med Mycol, 2003 Jun, 41(3), 235 - 9 Susceptibility profile of 200 bloodstream isolates of Candida spp . collected from Brazilian tertiary care hospitals; Colombo AL et al.; We evaluated the antifungal susceptibility profile of 200 recent bloodstream isolates of Candida spp . sequentially obtained from patients admitted to five tertiary care hospitals in Brazil . Isolates were identified by classical methods and the antifungal susceptibility profile was determined by the NCCLS microbroth assay method . Candida albicans was the most frequent species (41.5%); followed by C . tropicalis (24%) and C . parapsilosis (20.5%) . The frequency of C . glabrata and C . krusei was low (nine and two isolates, respectively) . Only three strains were resistant to fluconazole (two C . krusei and one C . glabrata) and only one was resistant to itraconazole (the same C . glabrata strain that was resistant to fluconazole) . Two strains were considered susceptible dose-dependent (SDD) to fluconazole and 13 isolates (6.5%) were SDD to itraconazole . Overall, the MIC50 value of non-C . albicans isolates for fluconazole was two dilutions higher than that of C . albicans isolates, and for itraconazole was one dilution higher . Resistance to amphotericin B (MIC > or = 2 microg ml(-1)) was observed in 2.5% of isolates (two strains of C . albicans, two of C . parapsilosis and one of C . krusei) . This study showed that episodes of candidemia in Brazilian public hospitals are represented mainly by fluconazole-susceptible non-C . albicans species . This finding is probably related to the low use of fluconazole in these hospitals. Dtsch Med Wochenschr, 2003 Sep 5, 128(36), 1829 - 32 {Diarrhea after vascular reconstruction of an abdominal aortic aneurysm}; Meibaum C et al.; HISTORY AND CLINICAL FINDINGS: A 60-year-old man had a dacron aortofemoral bypass graft inserted to replace a ruptured infrarenal aortic aneurysm rupture . He subsequently had tachyarrhythmic atrial fibrillation with heart failure, NYHA class IV, and diffuse abdominal pain associated with watery diarrhea . INVESTIGATIONS: Stool tests merely demonstrated Candida albicans . Abdominal ultrasound revealed intestinal loops with thickened walls and decreased peristalsis . Coloscopy demonstrated a retroperitoneal intestinal perforation with abscess formation resulting from ulcerative necrotizing rectosigmoid colitis which had also uncovered the vascular prosthesis near the abscess cavity . DIAGNOSIS: Ischemic transmural necrotizing rectosigmoiditis with retroperitoneal intestinal perforation . TREATMENT AND COURSE: The rectosigmoid colon was resected and an end-colostomy made with closure of the rectal stump (Hartmann's operation) . The uncovered right limb of the vascular graft was covered completely and was discharged, being now mobile using a walking frame . There was no evidence of infection in the dacron prosthesis . CONCLUSION: A transmural progression of an ischemic colitis should be considered as a late sequela after emergency vascular reconstruction of the abdominal aorta . Even if symptoms are mild, early postoperative sigmoidoscopy is indicated. J Ethnopharmacol, 2003 Oct, 88(2-3), 199 - 204 Antimicrobial activity of selected Peruvian medicinal plants; Rojas R et al.; The antimicrobial activity of 36 ethanol extracts from 24 plants, all of them currently used in the Peruvian traditional medicine for the treatment of several infectious and inflammatory disorders, was tested by means of the agar-well diffusion assay against four bacteria (Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa) and four fungi (Candida albicans, Trichophyton mentagrophytes, Microsporum gypseum and Sporothrix schenckii) . Twenty-five (69%) extracts showed some degree of antimicrobial activity against at least one microorganism . The plants with the greatest antimicrobial activity were Cestrum auriculatum L . Heritier (Solanaceae), Iryanthera lancifolia Ducke Suesseng (Myristicaceae), Lepechinia meyenii (Walp.) Epling (Lamiaceae) and Ophryosporus peruvianus (Gmelin) King & H . Rob . (Asteraceae). J Ethnopharmacol, 2003 Oct, 88(2-3), 189 - 93 Flavonoids and trypanocidal activity of Bulgarian propolis; Prytzyk E et al.; Acetone and ethanol extracts of two Bulgarian propolis samples (Bur and Lov) were investigated by high temperature high resolution gas chromatography coupled to mass spectrometry (HT-HRGC-MS), and their activity against Trypanosoma cruzi was evaluated . The ethanol extracts--Et-Bur and Et-Lov--showed similar composition, with a high content of flavonoids, and strong inhibitory activity against T . cruzi proliferative epimastigotes, which were more susceptible than trypomastigotes . In the presence of blood, the activity of Et-Bur or Et-Lov against trypomastigotes was similar to that of the standard drug, crystal violet . Both extracts also showed similar and significant activity against Staphylococcus aureus and Candida albicans, while being inactive against Escherichia coli . The acetone extract, Ket-Bur, was more active than Et-Bur against both forms of T . cruzi. J Ethnopharmacol, 2003 Oct, 88(2-3), 137 - 43 Osmitopsis asteriscoides (Asteraceae)-the antimicrobial activity and essential oil composition of a Cape-Dutch remedy; Viljoen A et al.; The essential oil composition and antimicrobial activity of Osmitopsis asteriscoides, a medicinal plant used in traditional herbal preparations in South Africa has been investigated . Three different antimicrobial methods (disc diffusion, minimum inhibitory concentration by micro-titer plate and time-kill studies) were comparatively evaluated against Candida albicans, Staphylococcus aureus and Pseudomonas aeruginosa . A preliminary screening was done using the disc diffusion method on nine bacterial and four fungal isolates . Minimum inhibitory concentrations showed some correlation with the disc diffusion method . However, time-kill studies appear to be a more superior method for determining antimicrobial activity of volatile compounds such as essential oils . Two moderately susceptible and one resistant organism were selected to further demonstrate the variability between the three methods . The antimicrobial activity of the essential oil, tested by means of time-kill methodology at concentrations ranging from 0.5 to 2% (v/v) indicate a strong fungicidal activity against Candida albicans and the oil was also found to be bacteriostatic against Staphylococcus aureus in a concentration-dependent manner . The essential oil rapidly reduced viable counts of Pseudomonas aeruginosa, but regrowth was noted after 240 min . The results have been generated in duplicate in separate microbiology laboratories using different time-kill methods and the results are congruent . The two major essential oil components camphor and 1,8-cineole were investigated indicating the positive antimicrobial efficacy of 1,8-cineole independently and in combination with camphor . In addition to (-)-camphor and 1,8-cineole, 40 compounds were identified by GC-MS in the hydro-distilled essential oil . The high concentration of cineole and camphor and their synergistic effect is presented as a possible explanation for the traditional use of Osmitopsis asteriscoides for treating microbe-related illnesses. Biochemistry, 2003 Sep 16, 42(36), 10822 - 32 Purification and characterization of the N-terminal nucleotide binding domain of an ABC drug transporter of Candida albicans: uncommon cysteine 193 of Walker A is critical for ATP hydrolysis; Jha S et al.; The Candida drug resistance protein Cdr1p (approximately 170 kDa) is a member of ATP binding cassette (ABC) superfamily of drug transporters, characterized by the presence of 2 nucleotide binding domains (NBD) and 12 transmembrane segments (TMS) . NBDs of these transporters are the hub of ATP hydrolysis activity, and their sequence contains a conserved Walker A motif (GxxGxGKS/T) . Mutations of the lysine residue within this motif abrogate the ability of NBDs to hydrolyze ATP . Interestingly, the sequence alignments of Cdr1p NBDs with other bacterial and eukaryotic transporters reveal that its N-terminal NBD contains an unusual Walker A sequence (GRPGAGCST), as the invariant lysine is replaced by a cysteine . In an attempt to understand the significance of this uncommon positioning of cysteine within the Walker A motif, we for the first time have purified and characterized the N-terminal NBD (encompassing first N-terminal 512 amino acids) of Cdr1p as well as its C193A mutant protein . The purified NBD-512 protein could exist as an independent functional general ribonucleoside triphosphatase with strong divalent cation dependence . It exhibited ATPase activity with an apparent K(m) in the 0.8-1.0 mM range and V(max) in the range of 147-160 nmol min(-)(1) (mg of protein)(-)(1) . NBD-512-associated ATPase activity was also sensitive to inhibitors such as vanadate, azide, and NEM . The Mut-NBD-512 protein (C193A) showed a severe impairment in its ability to hydrolyze ATP (95%); however, no significant effect on ATP (TNP-ATP) binding was observed . Our results show that C193 is critical for N-terminal NBD-mediated ATP hydrolysis and represents a unique feature distinguishing the ATP-dependent functionality of the ABC transporters of fungi from those found in bacteria and other eukaryotes. Yeast, 2003 Sep, 20(12), 1053 - 60 Identification of proteins highly expressed in the hyphae of Candida albicans by two-dimensional electrophoresis; Choi W et al.; The increase in Candida albicans infections is caused by the increase in therapies resulting in immunocompromised patients . One factor required for C . albicans pathogenicity is the morphological transition from yeast to hypha . The protein profiles of whole extracts from yeasts and hyphae were examined using two-dimensional electrophoresis to identify the proteins related to the morphological transition . Over 900 protein spots were visualized by silver staining and 11 spots were increased more than three-fold reproducibly during hyphal differentiation . Six of the 11 spots were identified by peptide mass fingerprints, of which three represented PRA1, two PHR1 and the last TSA1 . Vertical streak patterns of Pra1p and Phr1p indicated that post-translational modifications seem to be caused by variable glycosylation . Comparative proteome analysis between the wild-type and the deletion mutants, CAMB43 (deltapra1) and CAS10 (deltaphr1), further confirmed the identity of PRA1 and PHR1 . Interestingly, Pra1p was downregulated in phr1-deleted mutants . Only PHR1 transcription was increased, indicating that PRA1 and TSA1 are controlled at the post-translational level . Yeast, 2003 Sep, 20(12), 1045 - 52 Non-AUG translation initiation of mRNA encoding acidic ribosomal P2A protein in Candida albicans; Abramczyk D et al.; The eukaryotic 60S ribosomal subunit has a set of very acidic proteins (P-proteins), which form a distinct lateral protuberance called the stalk structure . This protein complex is directly involved in the elongation step of polypeptide synthesis . In our study on acidic ribosomal P-proteins from the human opportunistic pathogen Candida albicans, we isolated and characterized one of the genes, called CARP2A, and its product, the P2A protein . The CARP2A gene is intron-less, present in a single copy per haploid genome, and transcriptionally active . The open reading frame of the studied gene contains information for a sequence of 108 amino acids . Based on this, the molecular mass and isoelectric point of the P2A protein were theoretically calculated to be 10.85 kDa and 3.7, respectively . The characteristic feature of the CARP2A gene transcript is the presence of a GUG start codon, which is rare in eukaryotic organisms and not previously reported in yeast . To our knowledge this is the first report showing the presence of a naturally occurring non-AUG start codon on mRNA in yeast species . J Immunol, 2003 Sep 15, 171(6), 3047 - 55 Candida albicans induces selectively transcriptional activation of cyclooxygenase-2 in HeLa cells: pivotal roles of Toll-like receptors, p38 mitogen-activated protein kinase, and NF-kappa B; Deva R et al.; Candidiasis, in its mucocutaneous form as well as in an invasive form, is frequently associated with high morbidity . PGE(2), which is generated by enzymatic activity of cyclooxygenases (COXs) 1 and 2, has been shown to trigger morphogenesis in Candida albicans . In the present study, we investigated whether C . albicans altered COX-2 expression in HeLa cells . RT-PCR and Western blot analyses revealed a time-dependent biphasic behavior of COX-2 mRNA expression and COX-2 protein level . COX-1 protein remained unaffected . Neutralization with Abs against Toll-like receptors (TLR) 2 and 4 inhibited the Candida-induced production of PGE(2), suggesting a vital role for TLRs in the recognition and signaling in mammalian cells upon infection with C . albicans . Transient transfections with COX-2 promoter-luciferase construct and various inhibitors of mitogen-activated protein kinases (MAPK), such as protein kinase C (PKC) inhibitor GF203190X, p38(MAPK) inhibitor SB203109, and extracellular-regulated kinases 1 and 2 inhibitor PD98509 showed that C . albicans up-regulates selectively COX-2, but not COX-1, through p38(MAPK) and PKC pathways . No involvement of other stress kinases, e.g., c-Jun NH(2)-terminal kinase and extracellular-regulated kinases 1 and 2, was observed . Transient transfection of NF-kappaB promoter construct and dominant negative plasmid of IkappaBbeta kinase showed that COX-2 transcription is mediated through p38(MAPK) and NF-kappaB pathways . That NF-kappaB up-regulates p38(MAPK) is novel and is in contradiction to earlier reports in which NF-kappaB was shown to inhibit p38(MAPK) . In conclusion, multiple converging signaling pathways, involving TLRs followed by PKC, p38(MAPK), and/or NF-kappaB, are triggered by C . albicans in activation of COX-2 gene. Proc Natl Acad Sci U S A, 2003 Sep 16, 100(19), 11007 - 12 Epub 2003 Sep 04. Phagocytosis by neutrophils induces an amino acid deprivation response in Saccharomyces cerevisiae and Candida albicans; Rubin-Bejerano I et al.; The transcriptional profiles of yeast cells that have been phagocytosed by either human neutrophils or monocytes were compared by using whole genome arrays . After phagocytosis by neutrophils, both Saccharomyces cerevisiae and Candida albicans respond by inducing genes of the methionine and arginine pathways . Neither of these pathways is induced upon phagocytosis by monocytes . Both fungi show a similar induction of these pathways when transferred from amino acid-rich medium to amino acid-deficient medium . These data suggest that the internal phagosome of the neutrophil is an amino acid-deficient environment. Org Biomol Chem, 2003 Jul 21, 1(14), 2448 - 54 Stereochemistry of a bifunctional dihydroceramide delta 4-desaturase/hydroxylase from Candida albicans; a key enzyme of sphingolipid metabolism; Beckmann C et al.; The stereochemical course of the dihydroceramide delta 4-(E)-desaturase from Candida albicans, cloned and expressed in the yeast Saccharomyces cerevisiae strain sur2 delta, was determined using stereospecifically labelled (2R,3S)-{2,3,4,4-2H4}-palmitic acid as a metabolic probe . Mass spectrometric analysis of the dinitrophenyl-derivatives of the labelled long-chain bases revealed elimination of a single deuterium atom from C(4) (corresponding to the C(4)-HR) along with a hydrogen atom from C(5) (corresponding to the C(5)-HS) . This finding is consistent with an overall syn-elimination of the two vicinal hydrogen atoms . Besides the desaturation product sphingosine (93%) minor amounts of a 4-hydroxylated product (phytosphinganine, 7%) were identified that classify the Candida enzyme as a bifunctional desaturase/hydroxylase . Both processes, desaturation and hydroxylation proceed with loss of C(4)-HR from the chiral precursor . This finding is in agreement with a two-step process involving activation of the substrate by removal of the C(4)-HR to give a C-centred radical or radicaloid followed by either disproportionation into an olefin, water and a reduced diiron complex, or to recombination of the primary reactive intermediate with an active site-bound oxygen to yield a secondary alcohol . This result demonstrates the close mechanistic relationship between desaturation and hydroxylation as two different reaction pathways of a single enzyme and strengthens the mechanistic relationship of desaturases from fatty acid metabolism and sphingolipids. Chem Biol, 2003 Aug, 10(8), 743 - 50 Quorum sensing in Candida albicans: probing farnesol's mode of action with 40 natural and synthetic farnesol analogs; Shchepin R et al.; The dimorphic fungus Candida albicans produces extracellular farnesol (3,7,11-trimethyl-2,6,10-dodecatriene-1-ol) which acts as a quorum-sensing molecule (QSM) to suppress filamentation . Of four possible geometric isomers of farnesol, only the E,E isomer possesses QSM activity . We tested 40 natural and synthetic analogs of farnesol for their activity in an N-acetylglucosamine-induced differentiation assay for germ tube formation (GTF) . Modified structural features include the head group, chain length, presence or absence of the three double bonds, substitution of a backbone carbon by S, O, N, and Se heteroatoms, presence or absence of a 3-methyl branch, and the bulkiness of the hydrophobic tail . Of the 40 compounds, 22 showed QSM activity by their ability to reduce GTF by 50% . However, even the most active of the analogs tested had only 7.3% of the activity of E,E-farnesol . Structure-activity relationships were examined in terms of the likely presence in C . albicans of a farnesol binding receptor protein. Microbiol Immunol, 2003, 47(7), 479 - 90 Chemistry and biology of angiitis inducer, Candida albicans water-soluble mannoprotein-beta-glucan complex (CAWS); Ohno N; Deep mycoses have been clearly demonstrated to release beta-glucans into the blood . Structure of the beta-glucan was, at least in part, suggested to be a mannoprotein beta-glucan complex (CAWS) as assessed by biochemical and immunochemical analyses of the extracellular macromolecular fraction of Candida albicans . Half clearance time of i.v . administered CAWS was about 30 min in mice . In addition to the reactivity with limulus G-test, CAWS was found to exhibit various biological activities, such as cytokine synthesis by leukocyte, platelet aggregation, lethal toxicity, enhancement of side effect of indomethacin, induction of coronary arteritis in mice, and so on . In this review, the chemical properties and biological activities of CAWS are discussed. Genome Res, 2003 Sep, 13(9), 2005 - 17 Sixty alleles of the ALS7 open reading frame in Candida albicans: ALS7 is a hypermutable contingency locus; Zhang N et al.; The ALS (agglutinin-like sequence) gene family encodes proteins that play a role in adherence of the yeast Candida albicans to endothelial and epithelial cells . The proteins are proposed as virulence factors for this important fungal pathogen of humans . We analyzed 66 C . albicans strains, representing a worldwide collection of 266 infection-causing isolates, and discovered 60 alleles of the ALS7 open reading frame (ORF) . Differences between alleles were largely caused by rearrangements of repeat elements in the so-called tandem repeat domain (21 different types occurred) and the VASES region (19 different types) . C . albicans is diploid, and combinations of ALS7 alleles generated 49 different genotypes . ALS7 expression was detected in samples isolated directly from five oral candidosis patients . ORFs in the opposite direction contained within the ALS7 ORF were also transcribed in all strains tested . Isolates representing a more pathogenic general-purpose genotype (GPG) cluster of strains tended to have more tandem repeats than other strains . Two types of VASES regions were largely exclusive to GPG strains; the remaining types were largely exclusive to noncluster strains . Our results provide evidence that ALS7 is a hypermutable contingency locus and important for the success of C . albicans as an opportunistic pathogen of humans. Mycoses, 2003 Sep, 46(8), 307 - 11 Antifungal susceptibility testing of Candida albicans by flow cytometry; Gokahmetoglu S et al.; Antifungal susceptibilities of 28 Candida albicans isolates and two quality control strains to amphotericin B and fluconazole were determined by flow cytometry and microdilution method . Minimum inhibitory concentrations (MICs) obtained by flow cytometry were compared with the results obtained by The National Committee for Clinical Laboratory Standards Subcommittee (NCCLS) broth microdilution method . The agreement of results (within two dilution) obtained was found as 96 and 93% for amphotericin B and fluconazole, respectively . At least 24 h incubation was required for reading the microdilution assays . Four hours of incubation was required for fluconazole, whereas 2-h incubation was sufficient for amphotericin B to provide MIC by flow cytometry . Results of this study show that flow cytometry provides a rapid and sensitive in vitro method for antifungal susceptibility testing of Candida albicans isolates. Int Endod J, 2003 Sep, 36(9), 643 - 7 Examination on Candida spp . in refractory periapical granulomas; Waltimo T et al.; AIM: To examine the occurrence of Candida spp . in refractory periapical granulomas . METHODOLOGY: One hundred and three surgically removed periapical granulomas were subjected to molecular analysis for the occurrence of Candida albicans . DNA was extracted from the samples using a modified phenol/chloroform/isoamyl alcohol method and was subjected to polymerase chain reaction (PCR) with OPA-03 and repetitive sequence (GACA)4 primers . The PCR products were separated in agarose gel electrophoresis, stained with ethidium bromide, visualized using UV light and the sequences were analysed . Samples indicating possible occurrence of Candida were further investigated by histological and immunohistological methods . Periodic acid-Shiff staining (PAS) was used to detect yeast cells and hyphae, and specific monoclonal antibodies to recognize high molecular mass mannoproteins present in the C . albicans cell wall . DNA extraction was controlled by running PCR using beta-actin primers (a housekeeping gene) . C . albicans CCUG19915, C . tropicalis ATCC750, C . krusei ATCC6258, C . guilliermondii ATCC6260 and C . glabrata CCUG32725 served as positive controls in PCR . A tissue preparation of chronic atrophic candidosis in oral buccal mucosa served as a positive control for histological and immunohistological examinations . RESULTS: Polymerase chain reaction with beta-actin primers indicated successful DNA extraction in 68 out of 103 samples . The majority of the samples (50) were negative whereas 18 of the samples showed PCR products indicating possible occurrence of Candida spp . PAS-staining and immunohistological examination of these samples were, however, negative . Further analysis of the PCR products revealed sequences not typical for Candida spp . CONCLUSIONS: Candida spp . do not seem to occur in periapical granuloma. Microbiology, 2003 Sep, 149(Pt 9), 2597 - 608 N-acetylglucosamine-inducible CaGAP1 encodes a general amino acid permease which co-ordinates external nitrogen source response and morphogenesis in Candida albicans; Biswas S et al.; Candida albicans is able to grow in a variety of reversible morphological forms (yeast, pseudohyphal and hyphal) in response to various environmental signals, noteworthy among them being N-acetylglucosamine (GlcNAc) . The gene CaGAP1, homologous to GAP1, which encodes the general amino acid permease from Saccharomyces cerevisiae, was isolated on the basis of its induction by GlcNAc through differential screening of a C . albicans genomic library . The gene could functionally complement an S . cerevisiae gap1 mutant by rendering it susceptible to the toxic amino acid analogue mimosine in minimal proline media . As in S . cerevisiae, mutation of the CaGAP1 gene had an effect on citrulline uptake in C . albicans . Northern analysis showed that GlcNAc-induced expression of CaGAP1 was further enhanced in synthetic minimal media supplemented with single amino acids (glutamate, proline and glutamine) or urea (without amino acids) but repressed in minimal ammonium media . Induction of CaGAP1 expression by GlcNAc was nullified in C . albicans deleted for the transcription factor CPH1 and the hyphal regulator RAS1, indicating the involvement of Cph1p-dependent Ras1p signalling in CaGAP1 expression . A homozygous mutant of this gene showed defective hyphal formation in solid hyphal-inducing media and exhibited less hyphal clumps when induced by GlcNAc . Alteration of morphology and short filamentation under nitrogen-starvation conditions in the heterozygous mutant suggested that CaGAP1 affects morphogenesis in a dose-dependent manner. J Surg Res, 2003 Jul, 113(1), 42 - 9 Effect of lipopolysaccharide on virulence of intestinal candida albicans; Henry-Stanley MJ et al.; BACKGROUND: Candida albicans is a polymorphic fungus that frequently causes systemic infection in postsurgical and trauma patients . Others have reported that Escherichia coli lipopolysaccharide (LPS) acts as a copathogen to enhance the virulence of parenteral C . albicans . Experiments were designed to clarify the effect of parenteral LPS on systemic candidiasis initiated via the oral route . MATERIALS AND METHODS: Antibiotic-treated mice were orally inoculated with C . albicans CAF2 (wild-type) or mutant HLC54 (defective in filament formation), and were given 100 microg parenteral LPS 16 h before sacrifice . Separate groups of mice were additionally exposed to intermittent hypoxia prior to LPS . At sacrifice, cecal flora and microbial translocation to the mesenteric lymph nodes were quantified . C . albicans adherence to cultured HT-29 and Caco-2 enterocytes (pretreated with LPS, or calcium-free medium to expose the enterocyte lateral surface, or both) was quantified by enzyme-linked immunoabsorbent assay . RESULTS: All mice had high numbers of cecal C . albicans, and LPS was associated with an additional increase in cecal concentrations of HLC54 but not CAF2 . Translocation of HLC54, but not CAF2, appeared facilitated by hypoxia, but LPS did not facilitate translocation in any treatment group . Exposure of the lateral surface of cultured enterocytes had no effect on C . albicans adherence, although LPS consistently decreased adherence of both C . albicans strains . CONCLUSIONS: In contrast to experiments where systemic candidiasis was initiated by the parenteral route, parenteral LPS did not act as a copathogen in mice with systemic candidiasis initiated by the oral route, and these results might be related to LPS-induced alterations in C . albicans adherence to host enterocytes. Phytochemistry, 2003 Sep, 64(2), 543 - 8 Antifungal isopimaranes from Hypoestes serpens; Rasoamiaranjanahary L et al.; Five isopimarane diterpenes (7beta-hydroxyisopimara-8,15-dien-14-one, 14alpha-hydroxyisopimara-7,15-dien-1-one, 1beta,14alpha-dihydroxyisopimara-7,15-diene, 7beta-hydroxyisopimara-8(14),15-dien-1-one and 7beta-acetoxyisopimara-8(14),15-dien-1-one) have been isolated from the leaves of Hypoestes serpens (Acanthaceae) . All compounds exhibited antifungal activity against both the plant pathogenic fungus Cladosporium cucumerinum and the yeast Candida albicans; two of them also displayed an acetylcholinesterase inhibition . The structures of the compounds were determined by means of spectrometric methods, including 1D and 2D NMR experiments and MS analysis. Clin Infect Dis, 2003 Sep 1, 37(5), 634 - 43 Epub 2003 Aug 14. A prospective observational study of candidemia: epidemiology, therapy, and influences on mortality in hospitalized adult and pediatric patients; Pappas PG et al.; We conducted a prospective, multicenter observational study of adults (n=1447) and children (n=144) with candidemia at tertiary care centers in the United States in parallel with a candidemia treatment trial that included nonneutropenic adults . Candida albicans was the most common bloodstream isolate recovered from adults and children (45% vs . 49%) and was associated with high mortality (47% among adults vs . 29% among children) . Three-month survival was better among children than among adults (76% vs . 54%; P<.001) . Most children received amphotericin B as initial therapy, whereas most adults received fluconazole . In adults, Candida parapsilosis fungemia was associated with lower mortality than was non-parapsilosis candidemia (24% vs . 46%; P<.001) . Mortality was similar among subjects with Candida glabrata or non-glabrata candidemia; mortality was also similar among subjects with C . glabrata candidemia who received fluconazole rather than other antifungal therapy . Subjects in the observational cohort had higher Acute Physiology and Chronic Health Evaluation II scores than did participants in the clinical trial (18.6 vs . 16.1), which suggests that the former subjects are more often excluded from therapeutic trials. Curr Opin Microbiol, 2003 Aug, 6(4), 338 - 43 Adaptation of Candida albicans to the host environment: the role of morphogenesis in virulence and survival in mammalian hosts; Romani L et al.; Although morphological flexibility could be a key contributor to fungal virulence, no molecular data has unambiguously established fungal morphogenesis as a virulence factor for Candida albicans, nor can specific forms of Candida be regarded as absolutely indicative of saprophytism or infection at a given site on the host . The fitness of the fungus in vivo probably reflects its adaptation to the variety of microenvironments in which this opportunist must survive. Mol Microbiol, 2003 Sep, 49(5), 1391 - 405 CaSPA2 is important for polarity establishment and maintenance in Candida albicans; Zheng XD et al.; Saccharomyces cerevisiae Spa2p is a component of polarisome that controls cell polarity . Here, we have characterized the role of its homologue, CaSpa2p, in the polarized growth in Candida albicans . During yeast growth, GFP-tagged CaSpa2p localized to distinct growth sites in a cell cycle-dependent manner, while during hyphal growth it persistently localized to hyphal tips throughout the cell cycle . Persistent tip localization of the protein was also observed in Catup1Delta and Canrg1Delta, mutants constitutive for filamentous growth . Caspa2Delta exhibited defects in polarity establishment and maintenance, such as random budding and failure to confine growth to a small surface area leading to round cells with wide, elongated bud necks and markedly thicker hyphae . It was also defective in nuclear positioning, presumably a result of defective interactions between cytoplasmic microtubules with certain polarity determinants . The highly conserved SHD-I and SHD-V domains were found to be important and responsible for different aspects of CaSpa2p function . Caspa2Delta exhibited no virulence in the mouse systemic candidiasis model . Because of the existence of distinct growth forms and the easy control of the switch between them in vitro, C . albicans may serve as a useful model in cell polarity research. Pol Merkuriusz Lek, 2003 May, 14(83), 468 - 71 {Infectious non-venereal diseases affecting male and female external genital area}; Brzezinska-Wcislo L et al.; Infectious non-venereal diseases are divided into contagious and non-contagious skin diseases, that have sometimes different and unexplained aetiology . Some diseases caused by viruses, fungi, bacteria, Candida albicans and parasites are the contagious skin diseases . The non-contagious diseases include: contact dermatitis, erythema fixum, lichen sclerosus et atrophicus, Reiter's disease and some pre-neoplasmatic lesions . The aim of the study was to demonstrate some skin lesions commonly located on external genital organs in both sexes, that should be differentiated with skin lesions in sexually transmitted diseases. J Dent Res, 2003 Sep, 82(9), 748 - 52 Calcium blocks fungicidal activity of human salivary histatin 5 through disruption of binding with Candida albicans; Dong J et al.; Salivary histatin 5 (Hst 5) kills the fungal pathogen C . albicans via a mechanism that involves binding and subsequent efflux of cellular ATP . Our aims were to identify inorganic ions found in saliva that influence Hst 5 fungicidal activity . Increasing ionic strength with relevant salivary anions (Cl(-) and CO(3)(-)) did not reduce Hst 5 binding or uptake by yeast cells, but reduced the Hst-induced efflux of ATP . Extracellular MgCl(2) (25 mM) maximally inhibited 30-40% of Hst 5 killing with 40% reduction in ATP efflux, while pre-treatment of cells with only 2 mM CaCl(2) inhibited 80-90% of killing, and prevented ATP efflux . Loss of fungicidal activity by the addition of CaCl(2) or MgCl(2) was a result of inhibition of binding of Hst 5 to C . albicans cells . Calcium is a potent inhibitor of Hst 5 candidacidal activity at physiological concentrations and may be the primary salivary ion responsible for the masking effect of saliva. Eur J Clin Microbiol Infect Dis, 2003 Sep, 22(9), 558 - 60 Epub 2003 Aug 21. Rapid detection of pathogenic fungi from clinical specimens using LightCycler real-time fluorescence PCR; Imhof A et al.; In the study presented here a LightCycler real-time PCR system was used for the diagnosis of fungal infections from clinical tissue samples . Nine specimens were investigated from six patients with suspected or proven invasive fungal infections . Seven of nine samples were positive in a broad-range fungal PCR assay . In four samples, Aspergillus fumigatus was detected both by a species-specific hybridization assay as well as by sequencing of amplification products . In addition, the broad-range fungal PCR assay and PCR sequencing detected and identified, respectively, the following organisms in the specimens noted: Candida albicans in a culture-negative liver biopsy, Histoplasma capsulatum in a bone marrow sample, and Conidiobolus coronatus in a facial soft tissue specimen . Real-time PCR is a promising tool for the diagnosis of invasive fungal infections in human tissue samples and offers some advantages over culture methods, such as rapid analysis and increased sensitivity. Mem Inst Oswaldo Cruz, 2003 Jun, 98(4), 533 - 8 Epub 2003 Aug 18. Candida dubliniensis identification in Brazilian yeast stock collection; Mariano Pde L et al.; We investigated the presence of Candida dubliniensis among isolates previously identified as Candida albicans and maintained in a yeast stock collection from 1994 to 2000 . All isolates were serotyped and further evaluated for antifungal susceptibility profile . After doing a screening test for C . dubliniensis isolates based on the capability of colonies to grow at 42 C, its final identification was obtained by randomly amplified polymorphic DNA (RAPD) analysis using three different primers . A total of 46 out of 548 screened isolates did not exhibit growth at 42 C and were further genotyped by RAPD . Eleven isolates were identified as C . dubliniensis with RAPD analysis . Regarding serotypes, 81.5% of C . albicans and all C . dubliniensis isolates belonged to serotype A . Of note, 9 out of 11 C . dubliniensis isolates were obtained from patients with acquired immunodeficiency syndrome (Aids) and all of them were susceptible to azoles and amphotericin B . We found 17 (3%) C . albicans isolates that were dose-dependent susceptibility or resistant to azoles . In conclusion, we found a low rate of C . dubliniensis isolates among stock cultures of yeasts previously identified as C . albicans . Most of these isolates were recovered from oral samples of Aids patients and exhibited high susceptibility to amphotericin B and azoles . C . albicans serotype A susceptible to all antifungal drugs is the major phenotype found in our stock culture. Antimicrob Agents Chemother, 2003 Sep, 47(9), 2804 - 9 Antimicrobial characterization of human beta-defensin 3 derivatives; Hoover DM et al.; Human beta-defensin 3 (hBD3) is a highly basic 45-amino-acid protein that acts both as an antimicrobial agent and as a chemoattractant molecule . Although the nature of its antimicrobial activity is largely electrostatic, the importance of the molecular structure on this activity is poorly understood . Two isoforms of hBD3 were synthesized: the first with native disulfide linkages and the second with nonnative linkages . In a third synthetic peptide, all cysteine residues were replaced with alpha-aminobutyric acid, creating a completely linear peptide . A series of six small, linear peptides corresponding to regions of hBD3 with net charges ranging from +4 to +8 (at pH 7) and lengths ranging from 9 to 20 amino acids were also synthesized . The linear full-length peptide showed the highest microbicidal activity against Escherichia coli and Staphylococcus aureus, while all three full-length forms showed equal activity against Candida albicans . The linear peptide also showed high activity against Enterococcus faecium and Pseudomonas aeruginosa . Peptides corresponding to the C terminus showed higher activities when tested against E . coli, with the most active peptides being the most basic . However, only the peptide corresponding to the N terminus of hBD3 showed any activity against S . aureus and C . albicans . Further, N-terminal deletion mutants of native hBD3 showed diminished activities against S . aureus . Thus, the antimicrobial properties of hBD3 derivatives are determined by both charge and structure. Antimicrob Agents Chemother, 2003 Sep, 47(9), 2717 - 24 Disruption of ergosterol biosynthesis confers resistance to amphotericin B in Candida lusitaniae; Young LY et al.; Candida lusitaniae is an emerging human pathogen that, unlike other fungal pathogens, frequently develops resistance to the commonly used antifungal agent amphotericin B . Amphotericin B is a member of the polyene class of antifungal drugs, which impair fungal cell membrane integrity . Here we analyzed mechanisms contributing to amphotericin B resistance in C . lusitaniae . Sensitivity to polyenes in the related fungi Saccharomyces cerevisiae and Candida albicans requires the ergosterol biosynthetic gene ERG6 . In an effort to understand the mechanisms contributing to amphotericin B resistance in C . lusitaniae, we isolated the ERG6 gene and created a C . lusitaniae erg6 delta strain . This mutant strain exhibited a growth defect, was resistant to amphotericin B, and was hypersensitive to other sterol inhibitors . Based on the similarities between the phenotypes of the erg6 delta mutant and clinical isolates of C . lusitaniae resistant to amphotericin B, we analyzed ERG6 expression levels and ergosterol content in multiple clinical isolates . C . lusitaniae amphotericin B-resistant isolates were found to have increased levels of ERG6 transcript as well as reduced ergosterol content . These changes suggest that another gene in the ergosterol biosynthetic pathway could be mutated or misregulated . Further transcript analysis showed that expression of the ERG3 gene, which encodes C-5 sterol desaturase, was reduced in two amphotericin B-resistant isolates . Our findings reveal that mutation or altered expression of ergosterol biosynthetic genes can result in resistance to amphotericin B in C . lusitaniae. Infect Immun, 2003 Sep, 71(9), 5280 - 6 Differential role of MyD88 in macrophage-mediated responses to opportunistic fungal pathogens; Marr KA et al.; Toll-like receptors mediate macrophage recognition of microbial ligands, inducing expression of microbicidal molecules and cytokines via the adapter protein MyD88 . We investigated the role of MyD88 in regulating murine macrophage responses to a pathogenic yeast (Candida albicans) and mold (Aspergillus fumigatus) . Macrophages derived from bone marrow of MyD88-deficient mice (MyD88(-/-)) demonstrated impaired phagocytosis and intracellular killing of C . albicans compared to wild-type (MyD88(+/+)) macrophages . In contrast, ingestion and killing of A . fumigatus conidia was MyD88 independent . Cytokine production by MyD88(-/-) macrophages in response to C . albicans yeasts and hyphae was substantially decreased, but responses to A . fumigatus hyphae were preserved . These results provide evidence that MyD88 signaling is involved in phagocytosis and killing of live C . albicans, but not A . fumigatus . The differential role of MyD88 may represent one mechanism by which macrophages regulate innate responses specific to different pathogenic fungi. Infect Immun, 2003 Sep, 71(9), 4970 - 6 Skin facilitates Candida albicans mating; Lachke SA et al.; Mating between natural a/a and alpha/alpha strains of Candida albicans requires that cells first switch from the white to opaque phase phenotype . However, because cells expressing the opaque phase phenotype are induced to switch back to the white phase phenotype at physiological temperature (37 degrees C) and because opaque phase cells are highly efficient at colonizing skin, we tested whether skin, which is several degrees lower than physiological temperature, is conducive to mating . Using a model in which a mixture of a/a and alpha/alpha cells are incubated for 24 h under a cotton patch on the hairless skin of newborn mice and using scanning electron microscopy to visualize cells on skin, it was demonstrated that skin facilitates mating . In some regions of the skin, 40% of all cells had fused . All of the stages of mating observed in vitro were observed in vivo . However, some unique morphological characteristics of mating on skin were observed and are attributable to parent cell immobilization on the skin . In control experiments on glass, plastic, and silicone elastomer surfaces at 32 degrees C, cells either failed to fuse or did so at an extremely low frequency, suggesting that unique features of the skin surface other than reduced temperature also facilitate fusion. Eur J Obstet Gynecol Reprod Biol, 2003 Sep 10, 110(1), 66 - 72 An epidemiological survey of vulvovaginal candidiasis in Italy; Corsello S et al.; Eight Italian hospital or University gynecology clinics participated in a prospective survey of patients with culture-confirmed symptomatic vulvovaginal candidiasis (VVC) (October 1999 to March 2001) . Of 1138 patients recruited in the study, 931 were evaluable . A recent history of VVC was documented in 43.5% patients (358/823) with a mean number of 2.9+/-2.7 episodes per patient (N=302) . A total of 77 patients (10.0%) had a history of recurrent VVC (four and more episodes in a 12-month period) . The most frequent associated factors were related to life style: synthetic fabric underwear, vaginal douching and bike, training bike and motorbike (about 1/3 each) . Oral contraception was found in 20.8% patients, recent antibiotic use in 15.9% patients, current pregnancy concerned 10.3% patients while 3.4% patients were taking hormonal replacement therapy . Diabetes, corticosteroids or HIV were rarely encountered . Yeast was documented by direct microscopy in 78.3% patients (448/572) . A positive culture was obtained in 98.3% patients (909/925) . Candida albicans was the predominant species (77.1%), followed by Candida glabrata (14.6%) and Candida krusei (4.0%) . With the exception of one center with a lower proportion of C . albicans, this latter represented between 75 and 85% of the isolates . Overall, this study confirmed the preponderant role played by C . albicans in either sporadic and recurrent VVC. J Nat Prod, 2003 Aug, 66(8), 1132 - 5 Acetylenic acids inhibiting azole-resistant Candida albicans from Pentagonia gigantifolia; Li XC et al.; Antifungal bioassay-guided isolation of the ethanol extract of the roots of Pentagonia gigantifolia yielded 6-octadecynoic acid (1) and the new 6-nonadecynoic acid (2) . Compounds 1 and 2 inhibited the growth of fluconazole-susceptible and -resistant Candida albicans strains . Their antifungal potencies were comparable to those of amphotericin B and fluconazole . Of particular significance is the low cytotoxicity and specific activity of 1 and 2 against C . albicans. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2003 Aug, 96(2), 201 - 6 A new method for studying the adhesion of Candida albicans to dentin in the presence or absence of smear layer; Sen BH et al.; OBJECTIVES: The purpose of this study was to develop a reproducible, quantitative model of Candida albicans adhesion to human dentin through the use of a colorimetric method and to evaluate the effect of smear layer on candidal adhesion . STUDY DESIGN: Dentin disks with or without smear layer were incubated with C albicans (10(8) cells/mL) for 4 hours . After incubation, the disks were exposed to an (2,3)-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-{(phenylamino)-carbonyl}-2H-tetrazolium hydroxide-coenzyme Q solution for 2 hours . The color of (2,3)-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-{(phenylamino)-carbonyl}-2H-tetrazolium hydroxide formazan in the supernatant was determined spectrophotometrically at 492 nm . To relate formazan formation to cell numbers, standard curves were generated with known numbers of yeast cells without dentin . The number of adherent cells per square millimeter was then calculated . RESULTS: The number of attached C albicans cells was 2.4 x 10(4) per square millimeter in dentin with smear layer and 1.5 x 10(4) in dentin without smear layer (P <.05) . CONCLUSION: (2,3)-Bis(2-methoxy-4-nitro-5-sulfophenyl)-5-{(phenylamino)-carbonyl}-2H-tetrazolium hydroxide assay is a potential microbiologic tool for the quantitative determination of Candida adhesion to human dentin. Intervirology, 2003, 46(4), 199 - 206 Characterization of sialidase from an influenza A (H3N2) virus strain: kinetic parameters and substrate specificity; Franca de Barros J Jr et al.; Neuraminidase (NA) of influenza A (H3N2) viruses was characterized after purification by gel filtration and proteolytic treatment, using the X-31 variant strain that is a reassortment between the influenza A/Victoria/3/75 (responsible for the 1975 pandemic) and the influenza A/PR/8/34 virus samples, as a model . In the purification process, NA heads, that is the spike responsible for the virus sialidase activity, were purified by filtration through a Bio-Gel polyacrylamide column . The enzyme activity was determined by periodic acid/thiobarbituric acid assay and high-performance thin-layer chromatography . The sialidase showed preference for the alpha-2,3-linkage over the alpha-2,6-linkage of sialyllactoses (K(m) of 1.8 and 5.2 x 10(-4)M, respectively) at pH 5.2 . The enzyme acted on natural and synthetic substrates at different hydrolysis rates, as well as on human erythrocytes (A group, Rh+) and yeast (CANDIDA ALBICANS) cells . The active NA produced by gel filtration was characterized by different parameters of its sialidase activity, also showing to be a suitable tool for the identification of natural sialocompounds and for the screening of antisialidase drugs to treat influenza virus infections . Protein Sci, 2003 Sep, 12(9), 1822 - 32 Structure of the Escherichia coli malate synthase G:pyruvate:acetyl-coenzyme A abortive ternary complex at 1.95 A resolution; Anstrom DM et al.; Malate synthase, an enzyme of the glyoxylate pathway, catalyzes the condensation and subsequent hydrolysis of acetyl-coenzyme A (acetyl-CoA) and glyoxylate to form malate and CoA . In the present study, we present the 1.95 A-resolution crystal structure of Escherichia coli malate synthase isoform G in complex with magnesium, pyruvate, and acetyl-CoA, and we compare it with previously determined structures of substrate and product complexes . The results reveal how the enzyme recognizes and activates the substrate acetyl-CoA, as well as conformational changes associated with substrate binding, which may be important for catalysis . On the basis of these results and mutagenesis of active site residues, Asp 631 and Arg 338 are proposed to act in concert to form the enolate anion of acetyl-CoA in the rate-limiting step . The highly conserved Cys 617, which is immediately adjacent to the presumed catalytic base Asp 631, appears to be oxidized to cysteine-sulfenic acid . This can explain earlier observations of the susceptibility of the enzyme to inactivation and aggregation upon X-ray irradiation and indicates that cysteine oxidation may play a role in redox regulation of malate synthase activity in vivo . There is mounting evidence that enzymes of the glyoxylate pathway are virulence factors in several pathogenic organisms, notably Mycobacterium tuberculosis and Candida albicans . The results described in this study add insight into the mechanism of catalysis and may be useful for the design of inhibitory compounds as possible antimicrobial agents. Ir J Med Sci, 2003 Apr-Jun, 172(2), 60 - 2 Comparison of media for optimal recovery of Candida albicans and Candida glabrata from blood culture; Moore JE et al.; BACKGROUND: Candida spp., mainly Candida albicans, are frequently responsible for complications in immunocompromised patients . There are limited data comparing recovery efficiency using simple non-selective basal broth media . AIM: To compare several commercially available basal growth media to determine the medium that gave highest yeast proliferation . METHOD: Eight commercially available non-selective basal broth culture media were evaluated for optimum recovery of clinical C . albicans and C . glabrata . They included nutrient broth (NB), nutrient broth no . 2 (NB2), Todd-Hewitt (TH) broth, tryptone soya broth (TSB), tryptone soya broth supplemented with yeast extract (0.5% w/v {TSBYE}), brain heart infusion broth supplemented with yeast extract (0.5% w/v {BHIYE}), salt meat broth (SMB) and 0.1% {w/v} peptone saline (PS) . Differences in cell density were evaluated by spectrophotometrical analysis . RESULTS: TSBYE>BHIYE>TSB>TH>NB2>NB>SMB>PS for the optimum proliferation of cells in vitro . Either TSBYE or BHIYE broth may be employed as suitable basal broth media for growth of C . albicans and C . glabrata . NB should be considered the least suitable medium for routine use when others are available . CONCLUSION: These data may be of value to laboratories setting up simple blood culture systems to detect Candida spp., particularly in developing and underdeveloped countries. J Endod, 2003 Aug, 29(8), 501 - 4 Elimination of Candida albicans infection of the radicular dentin by intracanal medications; Siqueira JF Jr et al.; Fungi have been associated with cases of secondary or persistent root canal infections . The purpose of this study was to evaluate the effectiveness of four intracanal medications in disinfecting the root dentin of bovine teeth experimentally infected with Candida albicans . Infected dentin cylinders were exposed to four different medications: calcium hydroxide/glycerin; calcium hydroxide/0.12% chlorhexidine digluconate; calcium hydroxide/camphorated paramonochlorophenol/glycerin; and 0.12% chlorhexidine digluconate/zinc oxide . Specimens were in contact with the medications for 1 h, 2 days, and 7 days . The viability of C . albicans after exposure was evaluated by specimen incubation in culture medium to compare the effectiveness of the medications in disinfecting dentin . Results showed that the specimens treated with calcium hydroxide/camphorated paramonochlorophenol/ glycerin paste or with chlorhexidine/zinc oxide paste were completely disinfected after 1 h of exposure . Calcium hydroxide/glycerin paste only consistently eliminated C . albicans infection after 7 days of exposure . Calcium hydroxide mixed with chlorhexidine was ineffective in disinfecting dentin even after 1 week of medication exposure . Among the medications tested, the calcium hydroxide/camphorated paramonochlorophenol/glycerin paste and chlorhexidine digluconate mixed with zinc oxide were the most effective in eliminating C . albicans cells from dentinal specimens. Int J Antimicrob Agents, 2003 Aug, 22(2), 168 - 71 CAN-296-P is effective against cutaneous candidiasis in guinea pigs; Ben-Josef AM et al.; CAN-296 is a naturally occurring, heat-stable complex carbohydrate isolated from the cell wall of Mucor rouxii . Previously, CAN-296 demonstrated impressive in vitro fungicidal activity against a wide spectrum of pathogenic yeast, including azole-resistant isolates . The effect of CAN-296 on Candida albicans is rapid, concentration-dependent, and lethal . CAN-296-P is a chitosan-pyrithione derivative of CAN-296 containing 4% solution of chitosan with 25% substitution of pyrithione . Like CAN-296, it has in vitro fungicidal activity with a minimum inhibitory concentration (MIC) of 0.156 mg/l for C . albicans . The therapeutic effect of topical CAN-296-P on cutaneous candidiasis caused by C . albicans in guinea pigs was investigated . Three different C . albicans isolates, including one fluconazole-resistant strain (R 637601-9), were tested . After immunosuppression with cyclophosphamide, infection under occlusive dressing was achieved and treated within 48 h after the initial infection . Once-a-day topical application of 0.125, 0.25, 0.5, 1, 2, and 4% CAN-296-P solution was administrated for a period of 1, 3, 5, and 7 days . CAN-296-P at a concentration > or =0.25% was found to be as effective in clearing the infection as was 2% miconazole . Effectiveness in eradicating candidiasis with CAN-296-P was concentration-dependent and free of local adverse effects . Can-296-P is a novel, highly active topical fungicidal agent, with broad potential for clinical use. Gerodontology, 2003 Jul, 20(1), 50 - 6 A study on the effects of short-, medium- and long-term professional oral care in elderly persons requiring long-term nursing care at a chronic or maintenance stage of illness; Ueda K et al.; OBJECTIVE: The purpose of the present study is to clarify the frequency of provision of effective professional oral care by dental health workers in order to improve the oral hygiene state of elderly persons requiring long-term care . METHODS: In 105 patients requiring long-term nursing care, 55 subjects with positive oral Candida albicans were divided into five groups according to the frequency of oral care intervention at intervals of 1, 2, 3, 4 and 6 weeks . The effects of professional oral care on the improvement of the oral hygiene state were investigated taking the frequency and duration of care into consideration . RESULTS: The results showed that oral hygienic condition could be improved by performing professional oral care at intervals of 1 week for 12 consecutive weeks, and the improved condition could be maintained when professional oral care was continued at intervals of 1 week thereafter . CONCLUSION: It is necessary to continue professional oral care at intervals of 1 week for 12 consecutive weeks and at intervals of 2 weeks for more than 20 weeks . The oral hygiene state could be improved by professional intervention at intervals of three or four weeks in long-term oral care, but this was limited to subjects whose Functional Independence Measure (FIM) scores were higher than 3. Clin Microbiol Infect, 2003 Jul, 9(7), 684 - 90 Comparison of adherence of Candida albicans and Candida parapsilosis to silicone catheters in vitro and in vivo; Kojic EM et al.; OBJECTIVE: Although Candida parapsilosis has been associated with device-related infections in the clinical settings, factors that contribute to this association have not been previously examined . The objectives of this study were to compare in vitro and in vivo the adherence to silicone catheters of: (1) Candida albicans vs . C . parapsilosis, and (2) invasive vs . colonizing isolates of C . albicans and C . parapsilosis . METHODS: The records of 840 patients who had had Candida species isolated at three teaching hospitals during a three-month period were reviewed . A total of 20 clinical isolates of each of C . parapsilosis and C . albicans were examined for their adherence to silicone catheters in vitro and in a rabbit model of percutaneously placed catheters . For each Candida species, ten invasive isolates that had caused clinical device-related infection and 10 colonizing isolates that had caused only device colonization were studied . RESULTS: Candida parapsilosis accounted for <5% of yeast isolates from all sites, while three-quarters were C . albicans . Candida parapsilosis was isolated proportionately more often from blood and/or devices than C . albicans (34.3% vs . 8.5%, respectively, P < 0.0001) . There were no significant differences in the degrees of adherence in vitro and in vivo between C . albicans and C . parapsilosis or between invasive and colonizing Candida . CONCLUSION: Although C . parapsilosis was isolated proportionately more often from blood and/or devices than C . albicans in our studied population, there was no significant difference in the adherence of the two Candida species to silicone, nor between invasive and colonizing Candida in our in vitro and in vivo models . Factors other than microbial adherence may help explain the observed association of C . parapsilosis with device-related infections. J Coll Physicians Surg Pak, 2003 Aug, 13(8), 456 - 8 Candida albicans myocardial abscess; Bhatti MA et al.; Cardiac candidosis is a diagnostic and therapeutic challenge . This case report describes persistent candidemia which did not respond to standard therapy and the patient developed a myocardial abscess. AAPS PharmSciTech . 2002;3(4):E35. The development of Cutina lipogels and gel microemulsion for topical administration of fluconazole; El Laithy HM et al.; The influence of the vehicle on the release and permeation of fluconazole, a topical antifungal drug dissolved in Jojoba oil was evaluated . Series of Cutina lipogels (Cutina CPA {cetyl palmitate}, CBS {mixture of glyceryl stearate, cetearyl alcohol, cetyl palmitate, and cocoglycerides}, MD {glyceryl stearate}, and GMS {glyceryl monostearate}) in different concentrations as well as gel microemulsion were prepared . In-vitro drug release in Sorensen's citrate buffer (pH 5.5) and permeation through the excised skin of hairless mice, using a modified Franz diffusion cell, were performed . The rheological behavior and the apparent viscosity values for different gel bases were measured before and after storage under freezing conditions at -4 degrees C and were taken as measures for stability of network structure . Candida albicans was used as a model fungus to evaluate the antifungal activity of the best formula achieved . The results of in vitro drug release and its percutaneous absorption showed that the highest values from gel microemulsion were assured . The rheological behavior of the prepared systems showed pseudoplastic (shear-thinning) flow indicating structural breakdown of the existing intermolecular interactions between polymeric chains . Moreover, the stability study revealed no significant difference between viscosity before and after storage for different formulae except for CPA Cutina lipogel (using analysis of variance {ANOVA} test at level of significance.05) . The antifungal activity of fluconazole showed the widest zone of inhibition with gel microemulsion . The gel microemulsion is an excellent vehicle for fluconazole topical drug delivery. Eukaryot Cell, 2003 Aug, 2(4), 746 - 55 Roles of Candida albicans Dfg5p and Dcw1p cell surface proteins in growth and hypha formation; Spreghini E et al.; The Candida albicans cell wall participates in both growth and morphological transitions between yeast and hyphae . Our studies here focus on Dfg5p and Dcw1p, two similar proteins with features of glycosylphosphatidylinositol-linked cell surface proteins . Mutants lacking Dfg5p are defective in alkaline pH-induced hypha formation; mutants lacking Dcw1p have no detected hypha formation defect . Both homozygote-triplication tests and conditional expression strategies indicate that dfg5 and dcw1 mutations are synthetically lethal . Therefore, Dfg5p and Dcw1p share a function required for growth . Epitope-tagged Dfg5p, created through an insertional mutagenesis strategy, is found in cell membrane and cell wall extract fractions, and endoglycosidase H digestion shows that Dfg5p undergoes N-linked mannosylation . Surprisingly, Dfg5p is required for expression of the hypha-specific gene HWP1 in alkaline media . Because Dfg5p is a cell surface protein, it is poised to generate or transmit an external signal required for the program of hypha-specific gene expression. Eukaryot Cell, 2003 Aug, 2(4), 718 - 28 Diverged binding specificity of Rim101p, the Candida albicans ortholog of PacC; Ramon AM et al.; The biology of Candida albicans, including dimorphism and virulence, is significantly influenced by environmental pH . The response to ambient pH includes the pH-conditional expression of several genes, which is directly or indirectly regulated by Rim101p . Rim101p is homologous to PacC, a transcription factor that regulates pH-conditional gene expression in Aspergillus nidulans . PacC binds 5'-GCCARG-3' sequences upstream of pH-responsive genes and either activates or represses transcription . The absence of pacC consensus binding sites upstream of PHR1, a RIM101-dependent, alkaline pH-induced gene of C . albicans, suggested either that PHR1 is indirectly regulated by Rim101p or that the binding specificity of Rim101p is different . In vitro binding studies demonstrated that Rim101p strongly bound two regions upstream of PHR1 that were only weakly bound by PacC . Deletion analysis and site-specific mutagenesis demonstrated that both sites were functionally significant, mutation of either site reduced RIM101-dependent induction, and expression was abolished in the double mutant . Furthermore, oligonucleotides containing these sites conferred pH-conditional expression when inserted upstream of a reporter gene . The consensus sequence of these sites, 5'-CCAAGAAA-3', was identical to the binding recognition sequence identified by in vitro selection of Rim101p binding oligonucleotides from a random pool . The functional significance of this binding sequence was reinforced by its observed presence upstream of a number of newly identified pH-conditional genes . We conclude that Rim101p acts as a transcription factor and directly regulates pH-conditional gene expression but has a binding specificity different from that of PacC. Bull Exp Biol Med, 2003 May, 135(5), 460 - 3 IgE and IgG antibodies to Malassezia spp . yeast extract in patients with atopic dermatitis; Arzumanyan VG et al.; The presence of immunoglobulins to Malassezia spp . surface proteins in the sera from patients with atopic dermatitis and healthy subjects was studied . It was found that 28% of 25 examined patients with atopic dermatitis had IgE antibodies to Malassezia spp . surface protein preparation . All patients and 5 healthy subjects had IgG antibodies to this preparation . The presence and concentration of specific IgE antibodies in patients with atopic dermatitis correlated with reverse titers of IgG antibodies to this preparation (r=0.782) . The medians of values reciprocal to IgG antibody titers in patients with atopic dermatitis with and without specific IgE antibodies to the preparation and in healthy subjects were 64, 1024, and 16, respectively . The preparation derived from Candida albicans (candidine) and previously derived preparation from Malassezia did not cross-react . According to immunoblotting data, the preparation contains allergens presented by proteins with molecular weights 15, 36, 52-56, and 78.4 kDa. Crit Rev Oral Biol Med, 2003, 14(4), 253 - 67 Chronic hyperplastic candidosis/candidiasis (candidal leukoplakia); Sitheeque MA et al.; Chronic hyperplastic candidosis/candidiasis (CHC; syn . candidal leukoplakia) is a variant of oral candidosis that typically presents as a white patch on the commissures of the oral mucosa . The major etiologic agent of the disease is the oral fungal pathogen Candida predominantly belonging to Candida albicans, although other systemic co-factors, such as vitamin deficiency and generalized immune suppression, may play a contributory role . Clinically, the lesions are symptomless and regress after appropriate antifungal therapy and correction of underlying nutritional or other deficiencies . If the lesions are untreated, a minor proportion may demonstrate dysplasia and develop into carcinomas . This review outlines the demographic features, etiopathogenesis, immunological features, histopathology, and the role of Candida in the disease process . In the final part of the review, newer molecular biological aspects of the disease are considered together with the management protocols that are currently available, and directions for future research. Microbiol Immunol, 2003, 47(6), 395 - 403 Disruption of the human pathogenic yeast Candida albicans catalase gene decreases survival in mouse-model infection and elevates susceptibility to higher temperature and to detergents; Nakagawa Y et al.; Catalase-deficient strains of the human pathogenic yeast Candida albicans were constructed using the URA-blaster method . The disruptant was viable and grew normally in an ordinary culture condition, but became extremely sensitive to treatment with hydrogen peroxide . No catalase activity was observed in a catalase (CCT)-gene-disrupted strain, 1F5-4-1, suggesting that there were no other catalase or catalase-like enzymes in this yeast . The disruptant was shown to be sensitive to higher temperature and to low concentrations of SDS, NP-40, or Triton X-100 . After a wild-type CCT gene was reintroduced into the disruptant, catalase activity was restored and the strain became moderately sensitive to treatment with hydrogen peroxide . However, neither the temperature sensitivity nor the susceptibility to SDS observed in the disruptant was restored in the CCT-reintroduced strain . A model infection experiment using wild-type and dCCT strains showed that the disruptants disappeared more rapidly than the wild-type strain in mouse liver, lung, and spleen . These results suggest that the catalase plays a significant role in survival in the host immune system and thus leads this organism to establish infection in the host. Microbiology, 2003 Aug, 149(Pt 8), 2137 - 45 Identification and study of a Candida albicans protein homologous to Saccharomyces cerevisiae Ssr1p, an internal cell-wall protein; Garcera A et al.; After screening of a Candida albicans genome database, the product of an ORF (IPF 3054) that has 62 % homology with Saccharomyces cerevisiae Ssr1p, an internal cell-wall protein, was identified and named CaSsr1p . The deduced amino acid sequence shows that CaSsr1p contains an N-terminal hydrophobic signal peptide, is rich in Ser and Thr amino acids and has a potential glycosylphosphatidylinositol-attachment signal . CaSsr1p is released following degradation of isolated cell walls by zymolyase (mainly a 1,3-beta-glucanase) and therefore seems to be covalently linked to the beta-glucan of the cell walls . Both disruption and overexpression of the CaSSR1 gene caused an increased sensitivity to calcofluor white, Congo red and zymolyase digestion . These results suggest that CaSsr1p has a structural role associated with the cell-wall beta-glucan. J Clin Microbiol, 2003 Aug, 41(8), 3765 - 76 Optimization and validation of multilocus sequence typing for Candida albicans; Tavanti A et al.; Multilocus sequence typing (MLST) was applied to 75 Candida albicans isolates, including 2 that were expected to be identical, 48 that came from diverse geographical and clinical sources, and 15 that were sequential isolates from two patients . DNA fragments ( approximately 500 bp) of eight genes encoding housekeeping functions were sequenced, including four that have been described before for C . albicans MLST, and four new gene fragments, AAT1a, AAT1b, MPI, and ZWF1 . In total, 87 polymorphic sites were found among 50 notionally different isolates, giving 46 unique sequence types, underlining the power of MLST to differentiate isolates for epidemiological studies . Additional typing information was obtained by detecting variations in size at the transcribed spacer region of the 25S rRNA gene and tests for homozygosity at the mating type-like (MTL) locus . The stability of MLST was confirmed in two sets of consecutive isolates from two patients . In each set the isolates were identical or varied by a single nucleotide . Reference strain SC5314 and a derived mutant, CAF2, gave identical MLST types . Heterozygous polymorphisms were found in at least one isolate for all but 16 (18.4%) of the variable nucleotides, and 35 (41%) of the 87 individual sequence changes generated nonsynonymous amino acids . Cloning and restriction digestion of a gene fragment containing heterozygous polymorphisms indicated that the heterozygosity was genuine and not the result of sequencing errors . Our data validate and extend previous MLST results for C . albicans, and we propose an optimized system based on sequencing eight gene fragments for routine MLST with this species. Appl Environ Microbiol, 2003 Aug, 69(8), 4566 - 74 Rapid identification of Candida species by using nuclear magnetic resonance spectroscopy and a statistical classification strategy; Himmelreich U et al.; Nuclear magnetic resonance (NMR) spectra were acquired from suspensions of clinically important yeast species of the genus Candida to characterize the relationship between metabolite profiles and species identification . Major metabolites were identified by using two-dimensional correlation NMR spectroscopy . One-dimensional proton NMR spectra were analyzed by using a staged statistical classification strategy . Analysis of NMR spectra from 442 isolates of Candida albicans, C . glabrata, C . krusei, C . parapsilosis, and C . tropicalis resulted in rapid, accurate identification when compared with conventional and DNA-based identification . Spectral regions used for the classification of the five yeast species revealed species-specific differences in relative amounts of lipids, trehalose, polyols, and other metabolites . Isolates of C . parapsilosis and C . glabrata with unusual PCR fingerprinting patterns also generated atypical NMR spectra, suggesting the possibility of intraspecies discontinuity . We conclude that NMR spectroscopy combined with a statistical classification strategy is a rapid, nondestructive, and potentially valuable method for identification and chemotaxonomic characterization that may be broadly applicable to fungi and other microorganisms. Bioorg Med Chem, 2003 Aug 15, 11(17), 3709 - 22 Rebeccamycin analogues bearing amine substituents or other groups on the sugar moiety; Anizon F et al.; In the course of structure-activity relationship studies on rebeccamycin analogues, a series of compounds bearing an amino function on the sugar moiety were synthesized with the aim of improving the solubility and interaction with the macromolecular target(s) . The syntheses of amino derivatives and the corresponding chloro, iodo and azido intermediates are described . Their interaction with DNA and effects on human DNA topoisomerases I and II were investigated . Their antimicrobial activities against two Gram-positive bacteria, Bacillus cereus and Streptomyces chartreusis, a Gram-negative bacterium Escherichia coli and a yeast Candida albicans were also determined . 6'-Amino compound 7 and 6'-N-methylamino 14 very efficiently inhibit the growth of E . coli . The introduction of an amino group at the 6'-position strongly enhances the capacity of the drugs to interact with DNA but almost abolishes their poisoning effect on topoisomerase I . Unlike the vast majority of rebeccamycin analogues previously studied, the newly designed compounds do not stimulate DNA cleavage by topoisomerase I . The enhanced capacity of the 6'-amino glycosyl rebeccamycin derivatives to bind to DNA likely account for the improved biological profiles . DNA and topoisomerase I represent two independent targets which can both be used for the development of antitumor rebeccamycin derivatives. Yeast, 2003 Aug, 20(11), 929 - 41 Protective roles of mitochondrial manganese-containing superoxide dismutase against various stresses in Candida albicans; Hwang CS et al.; Candida albicans contains copper- and zinc-containing superoxide dismutase but also two manganese-containing superoxide dismutases (MnSODs), one in the cytosol and the other in the mitochondria . Among these, the SOD2 gene encoding mitochondrial MnSOD was disrupted and overexpressed to investigate its roles in C . albicans . The null mutant lacking mitochondrial MnSOD was more sensitive than wild-type cells to various stresses, such as redox-cycling agents, heating, ethanol, high concentration of sodium or potassium and 99.9% O2 . Interestingly, the sod2/sod2 mutant was rather more resistant to lithium and diamide than the wild-type, whereas overexpression of SOD2 increased susceptibility of C . albicans to these compounds . The inverse effect of mitochondrial MnSOD on lithium toxicity was relieved when the sod2/sod2 and SOD2-overexpressing cells were grown on the synthetic dextrose medium containing sulphur compounds such as methionine, cysteine, glutathione or sulphite, indicating that mitochondrial MnSOD may affect lithium toxicity through sulphur metabolism . Moreover, disruption or overexpression of SOD2 increased or decreased glutathione reductase activity and cyanide-resistant respiration by alternative oxidase, respectively . Taken together, these findings suggest that mitochondrial MnSOD is important for stress responses, lithium toxicity and cyanide-resistant respiration of C . albicans . J Infect Dis, 2003 Aug 15, 188(4), 591 - 9 Epub 2003 Jul 24. Hydrolytic gene expression during oroesophageal and gastric candidiasis in immunocompetent and immunodeficient gnotobiotic mice; Schofield DA et al.; To investigate whether host immunocompetence influences hydrolytic gene expression, we compared secretory aspartyl proteinase gene (SAP) and phospholipase B gene (PLB) expression during gastric candidiasis in immunocompetent and defined immunodeficient gnotobiotic mice, by reverse-transcription polymerase chain reaction . The use of immunodeficient gnotobiotic mice with combined defects in T cells and natural killer cells enabled a comprehensive study of virulence gene expression in various mucosal sites during lethal oroesophageal (tongue, palate, and esophagus) and gastric candidiasis . All 10 SAP and both PLB genes were expressed in both immunocompetent and specific immunodeficient mice, which suggests that the absence of important components of the host defense did not alter gene expression during gastric candidiasis . Although similar patterns of gene expression were evident in different oral tissues, we detected specific differences between Candida albicans-infected oroesophageal and gastric tissues and differences at various time points during the progression of gastric candidiasis. Can J Microbiol, 2003 May, 49(5), 344 - 9 Modulatory effect of cAMP on fungal ergosterol level and inhibitory activity of azole drugs; Sardari S et al.; The functions and biosynthesis of sterols have been effective targets for fungal control in different areas, including pharmaceutical and agricultural applications . Fungi are among the organisms that synthesize sterols, principally ergosterol . In this paper, the effect of dibutyryl-cAMP (db-cAMP) on ergosterol level and the interaction of drugs that would change the concentration of cAMP with antifungal drugs have been investigated . Sterols were extracted from Candida albicans, and ergosterol was measured using the gas chromatography method . The interaction of different agents was measured by the broth dilution method . It was found that phosphodiesterase inhibitors reverse the inhibitory activity of azole antifungal drugs . Evaluating the ergosterol level of C . albicans incubated with db-cAMP revealed that it increased ergosterol level . Further experiments provided evidence attributing the observed interaction between azoles and phosphodiesterase inhibitors to the relationship between ergosterol and cAMP . The possible significance of this interaction includes potentiation of antifungal activity of drugs by manipulating the cAMP level. J Am Acad Dermatol, 2003 Aug, 49(2), 187 - 92 Superficial fungal infections in 102 renal transplant recipients: a case-control study; Gulec AT et al.; BACKGROUND: Renal transplant recipients are predisposed to superficial fungal infections caused by graft-preserving immunosuppressive therapy . Reports have documented a wide range of prevalence rates for superficial fungal infections in this patient group . OBJECTIVE: The aim of this study was to determine the prevalence and clinical and mycological features of superficial fungal infections in renal transplant recipients at our center . METHODS: One hundred two consecutively registered renal transplant recipients (34 women, 68 men) and 88 healthy age- and sex-matched persons acting as controls (30 women, 58 men) underwent screening for the presence of superficial fungal infection . Skin scrapings and swabs were obtained from the dorsum of the tongue, upper part of the back, toe webs, and any suspicious lesions . Nail clippings were also collected . All samples were examined by direct microscopy and were stained with calcofluor white . The samples were cultured in Sabouraud dextrose agar, mycobiotic agar, and dermatophyte test medium . Candida species were identified on the basis of germ-tube production, spore formation in cornmeal agar, and results of biochemical testing . Dermatophytes were identified on the basis of colonial and microscopic morphologic features in conjunction with results of physiologic evaluation (in vitro hair perforation test, urease activity, temperature tolerance test, and nutritional test) . RESULTS: Sixty-five (63.7%) of the 102 renal transplant recipients had cutaneous-oral candidiasis, dermatophytosis, or pityriasis versicolor, whereas only 27 (30.7%) of controls had fungal infection . Pityriasis versicolor was the most common fungal infection in the patient group (36.3%), followed by cutaneous-oral candidiasis (25.5%), onychomycosis (12.7%), and fungal toe-web infection (11.8%) . Pityriasis versicolor and oral candidiasis were significantly more common among the renal transplant recipients, whereas the frequency of dermatophytosis in patients and controls was similar . Candida albicans was the main agent responsible for oral candidiasis, and Trichophyton rubrum was the most common dermatophyte isolated . Analysis showed that age, sex, and duration of immunosuppression did not significantly affect the prevalence of superficial fungal infection . Cyclosporine treatment and azathioprine therapy were identified as independent risk factors for superficial fungal disease . CONCLUSIONS: The prevalence of opportunistic infections with Pityrosporum ovale and C albicans is increased among renal transplant recipients, probably owing to the immunosuppressed state of this patient population . However, renal transplant recipients are not at increased risk of dermatophytosis. Chemosphere, 2003 Oct, 53(1), 71 - 7 Disinfection of surfaces by photocatalytic oxidation with titanium dioxide and UVA light; Kuhn KP et al.; Particularly in microbiological laboratories and areas in intensive medical use, regular and thorough disinfection of surfaces is required in order to reduce the numbers of bacteria and to prevent bacterial transmission . The conventional methods of disinfection with wiping are not effective in the longer term, cannot be standardized, are time- and staff-intensive and use aggressive chemicals . Disinfection with hard ultraviolet C (UVC) light is usually not satisfactory, as the depth of penetration is inadequate and there are occupational medicine risks . Photocatalytic oxidation on surfaces coated with titanium dioxide (TiO2) might offer a possible alternative . In the presence of water and oxygen, highly reactive OH-radicals are generated by TiO2 and mild ultraviolet A (UVA) . These radicals are able to destroy bacteria, and may therefore be effective in reducing bacterial contamination . Direct irradiation with UVC however can produce areas of shadow in which bacteria are not inactivated . Using targeted light guidance and a light-guiding sheet (out of a UVA-transmittant, Plexiglas, for example), as in the method described in the present study, bacterial inactivation over the entire area is possible . The effectiveness of the method was demonstrated using bacteria relevant to hygiene such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecium . For these bacteria, a reduction efficiency (RE) more than 6log10 steps in 60 min was observed . Using Candida albicans, a RE of 2log10 steps in 60 min was seen . Light and scanning electron microscopic examinations suggest that the germ destruction achieved takes place through direct damage to cell walls caused by OH-radicals. J Bone Joint Surg Br, 2003 Jul, 85(5), 734 - 5 Candida septic arthritis of the hip in a young patient without predisposing factors; Kawanabe K et al.; We present a case of septic arthritis of the hip caused by Candida albicans in a 24-year-old woman . This is the first report of a Candida infection of the hip in a patient without any predisposing factors . She underwent a two-stage total hip arthroplasty three years after the onset of the infection . An elevated plasma level of beta-D-glucan suggested Candida as the infecting organism and the diagnosis might have been made earlier if this test had been undertaken at presentation. Mol Pathol, 2003 Aug, 56(4), 232 - 6 Isolation and partial characterisation of a new antiproliferative substance from human leucocytes inhibiting growth of Candida albicans; Naess-Andresen CF et al.; AIM: To purify and partially characterise a fraction from human leucocytes containing a substance cytotoxic to Candida albicans . METHODS: Leucocytes were isolated from the buffy coats of healthy blood donors . The cytotoxic factor (CF) was isolated from the soluble fraction of the cells . A cell lysate was passed through a filter with a cut off value of 3 kDa, and the filtrate was processed by anionic exchange chromatography and gel filtration . The purified CF was analysed for its chemical and biological properties . The cytotoxicity of CF was tested on C albicans grown on agar plates . RESULTS: Mass spectrometry showed a molecular mass of 2.148 kDa . CF was found in polymorphonuclear neutrophilic cells only . No amino acids were detected, and a low ultraviolet absorbance at 260 nm and resistance to nuclease indicate the absence of nucleic acids . An anthrone test was positive for carbohydrate . The substance was soluble in water . CF showed a dose related cytotoxicity in the range of 0.1-1 mg/ml . The cytotoxic effect was abrogated by zinc ions . Preliminary testing indicated that CF also had cytotoxic effects against some bacteria . CONCLUSIONS: This report describes a factor from isolated human leucocytes that is cytotoxic to C albicans . The substance contains a carbohydrate moiety, whereas no amino acids were detected . The cytotoxicity can be abrogated by zinc ions in vitro . This substance is probably part of the repertoire by which leucocytes prevent infections. Chem Biol, 2003 Jul, 10(7), 635 - 44 DNA binding hairpin polyamides with antifungal activity; Marini NJ et al.; Eight-ring hairpin polyamides containing N-methylimidazole (Im) and N-methylpyrrole (Py) amino acids have been shown to bind with subnanomolar affinity to discrete DNA sites and to modulate a variety of DNA-dependent biological processes . We show here that addition of a second positive charge at the C terminus of an 8-ring hairpin polyamide confers activity against a number of clinically relevant fungal strains in vitro, and activity against Candida albicans in a mouse model . Control experiments indicate that the observed antifungal activity results from a DNA binding mechanism-of-action that does not involve DNA damage or disruption of chromosomal integrity . Hairpin activity is shown to be proportional to yeast DNA content (ploidy) . Transcriptional interference is proposed as the likely explanation for fungal cytotoxicity . Experiments with sensitized yeast strains indicate the potential for discrete sites of action rather than global effects. J Antimicrob Chemother, 2003 Sep, 52(3), 419 - 27 Epub 2003 Jul 29. Polymeric carriers for amphotericin B: in vitro activity, toxicity and therapeutic efficacy against systemic candidiasis in neutropenic mice; Espuelas MS et al.; OBJECTIVE: To study the toxicity and activity of two new amphotericin B formulations: poly(epsilon-caprolactone) nanospheres coated with poloxamer 188 (AmB-NP) and mixed micelles with the same surfactant (AmB-MM) . MATERIALS AND METHODS: The toxicity of these formulations was evaluated in erythrocytes, J774.2 macrophages and LLCPK1 renal cells, as well as in mice . Activity was determined in clinical isolates and in neutropenic mice . Mice were made neutropenic with 5-fluorouracil, infected with Candida albicans and treated with the antifungal formulations for three consecutive days . AmB association in cells and accumulation in kidneys and liver of animals was quantified by HPLC . RESULTS: Both formulations decreased between 8- and 10-fold the MIC of the polyene against clinical isolates of C . albicans . However, their activity was lower than or equal to that of AmB-deoxycholate when it was assessed against C . albicans-infected macrophages . When given as a single intravenous dose in mice, AmB-MM and AmB-NP had an LD50 of 9.8 and 18.6 mg/kg, respectively, compared with 4 mg/kg for AmB-deoxycholate . Comparison of residual infection burdens in the liver and kidneys showed that AmB-deoxycholate (0.5 mg/kg) was more effective and faster in eradicating yeast cells than polymeric formulations . This fact can be related to a lower AmB accumulation inside macrophages and in liver and kidneys (about 1.5 mg drug/g tissue) of mice, compared with those detected for AmB-deoxycholate (4 mg drug/g) . Overall, the efficacy of these formulations at 2 mg/kg was equal to that of AmB-deoxycholate at 0.5 mg/kg . CONCLUSIONS: AmB-MM and AmB-NP decreased the in vivo antifungal activity of AmB, and higher concentrations were therefore necessary to obtain a similar therapeutic effect . However, these higher concentrations were achievable owing to the reduced toxicity of these formulations. J Leukoc Biol, 2003 Aug, 74(2), 206 - 15 Mannose receptor contribution to Candida albicans phagocytosis by murine E-clone J774 macrophages; Porcaro I et al.; Mannoproteins, as the main constituents of the outer layer of yeast cell walls, are able to interact with phagocytic cells in an opsonin-independent manner through the mannose receptor (MR) and to induce yeast ingestion by the professional phagocytes . Moreover, the MR also mediates endocytosis of soluble ligands through clathrin-coated pits . Here, we studied some aspects of the interaction between the MR and Candida albicans using murine E-clone macrophages and the consequences on MR trafficking . Using a pull-down assay involving mixture E-clone macrophage detergent lysate with mannosylated Sepharose beads and glutaraldehyde-fixed, heat-killed (HK) C . albicans, we found that binding of solubilized MR to mannosylated particles occurred with characteristics similar to the receptor's cell-surface mannose-binding activity . We then demonstrated that MR expressed on E-clone macrophages contributed to phagocytosis of unopsonized, HK C . albicans and that yeast phagocytosis induced a decrease in MR endocytic activity without concomitant degradation of the receptor i