Folia Microbiol (Praha), 2000, 45(2), 138 - 42 Antifungal properties of substituted 1-phenyl-5-mercaptotetrazoles and their oxidation product, 5-bis-(1-phenyltetrazolyl)disulfide; Nesmerak K et al.; The antifungal effect of substituted 1-phenyl-5-mercaptotetrazoles was tested with Candida tropicalis, C . pseudotropicalis, C . mogii, Trichosporon cutaneum, Cryptococcus albidus and S . cerevisiae . Candida strains exhibited the lowest sensitivity to the compounds; the most sensitive was S . cerevisiae . The MIC values ranged from 40 to > 1000 mg/mL . The antifungal effect of halogenated compounds decreased in the series of bromo > chloro > fluoro derivatives . The electrochemical oxidation of substituted 1-phenyl-5-mercaptotetrazole derivatives in an acetonitrile medium was studied as a model for the enzymic oxidation of the substance, including study of the effect of water, perchloric and trifluoromethanesulfuric acids on E1/2 and I1 . 5-Bis-(1-phenyltetrazolyl)disulfide, the compound with no antifungal effect, has been identified as the main oxidation product of 1-phenyl-5-mercaptotetrazole. Folia Microbiol (Praha), 2000, 45(3), 255 - 8 Isolation and identification of xylitol-producing yeasts from agricultural residues; Altamirano A et al.; Selected yeast strains isolated from corn silage and viticulture residues were screened for their capacities to convert D-xylose into xylitol A conventional TLC was adapted for easy determination of xylose and xylitol in the culture supernatant solutions . This technique is suitable for the first steps of a screening program to select xylitol-producing yeasts from natural environments . Candida tropicalis ASM III (NRRL Y-27290), isolated from corn silage, appears to be a promising strain for xylitol production with a high yield (0.88 g xylitol per g of xylose consumed). Lett Appl Microbiol, 2001 Mar, 32(3), 190 - 3 Optimum conditions for yeast protoplast release and regeneration in Saccharomyces cerevisiae and Candida tropicalis using gut enzymes of the giant African snail Achatina achatina; Ezeronye OU et al.; Release of viable protoplasts of Saccharomyces cerevisiae and Candida tropicalis was achieved using fresh crude enzyme extracts of the giant African snail Achatina achatina . Optimum results of 2.8 x 10(6) protoplast ml(-1) were obtained when 1 g (wet wt) of cell slurry from the yeast strains was first treated with 1% beta-mercaptoethanol for 10 min and incubated with the undiluted crude enzyme for 120 min using 1.0 mol l(-1) sorbitol as osmotic stabilizer . Protoplast yield was enhanced with higher enzyme concentrations, longer digestion times and treatment of cells with beta-mercaptoethanol . Percentage regeneration of protoplast to viable cells in isotonic medium containing 0.01 mol l(-1) CaCl2 was in the range of 52-77% . These findings could be useful in the genetic manipulation of yeast of industrial importance. Bioresour Technol, 2001 Mar, 77(1), 57 - 63 Production of ethanol and xylitol from corn cobs by yeasts; Latif F et al.; Saccharomyces cerevisiae and Candida tropicalis were used separately and as co-culture for simultaneous saccharification and fermentation (SSF) of 5-20% (w/v) dry corn cobs . A maximal ethanol concentration of 27, 23, 21 g/l (w/v) from 200 g/l (w/v) dry corn cobs was obtained by S . cerevisiae, C . tropicalis and the co-culture, respectively, after 96 h of fermentation . However, theoretical yields of 82%, 71% and 63% were observed from 50 g/l dry corn cobs for the above cultures, respectively . Maximal xylitol concentration of 21, 20 and 15 g/l from 200 g/l (w/v) dry corn cobs was obtained by C . tropicalis, co-culture, and S . cerevisiae, respectively . Maximum theoretical yields of 79.0%, 77.0% and 58% were observed from 50 g/l of corn cobs, respectively . The volumetric productivities for ethanol and xylitol increased with the increase in substrate concentration, whereas, yield decreased . Glycerol and acetic acid were formed as minor by-products . S . cerevisiae and C . tropicalis resulted in better product yields (0.42 and 0.36 g/g) for ethanol and (0.52 and 0.71 g/g) for xylitol, respectively, whereas, the co-culture showed moderate level of ethanol (0.32 g/g) and almost maximal levels of xylitol (0.69 g/g). Bioresour Technol, 2001 Feb, 76(3), 213 - 20 The influence of aeration and hemicellulosic sugars on xylitol production by Candida tropicalis; Walther T et al.; The influence of other hemicellulosic sugars (arabinose, galactose, mannose and glucose), oxygen limitation, and initial xylose concentration on the fermentation of xylose to xylitol was investigated using experimental design methodology . Oxygen limitation and initial xylose concentration had considerable influences on xylitol production by Canadida tropicalis ATCC 96745 . Under semiaerobic conditions, the maximum xylitol yield was 0.62 g/g substrate, while under aerobic conditions, the maximum volumetric productivity was 0.90 g/l h . In the presence of glucose, xylose utilization was strongly repressed and sequential sugar utilization was observed . Ethanol produced from the glucose caused 50% reduction in xylitol yield when its concentration exceeded 30 g/l . When complex synthetic hemicellulosic sugars were fermented, glucose was initially consumed followed by a simultaneous uptake of the other sugars . The maximum xylitol yield (0.84 g/g) and volumetric productivity (0.49 g/l h) were obtained for substrates containing high arabinose and low glucose and mannose contents. Phytomedicine, 2000 Jun, 7(3), 235 - 8 Bioactivity studies of extracts from Tridax procumbens; Taddei A et al.; An updated review on the biological activity of Tridax procumbens is presented . A detailed biological screening comprised of gram-positive and gram-negative bacteria, yeasts and fungi using crude extracts of this plant was undertaken . The n-hexane extract of the flowers showed activity against Escherichia coli . The same extract of the whole aerial parts was active against Mycobacterium smegmatis, Escherichia coli, Salmonella group C and Salmonella paratyphi . The ethyl-acetate extract of the flowers was active against Bacillus cereus and Klebsiella sp . The aerial parts extract also showed activity only against Mycobacterium smegmatis and Staphylococcus aureus, while the aqueous extract showed no antimicrobial activity . None of the tested extracts was active against the yeasts, Candida albicans, Candida tropicalis and Rhodotorula rubra; or the fungi: Aspergillus flavus, Aspergillus niger, Mucor sp . and Trichophyton rubrum. Exp Eye Res, 2001 Feb, 72(2), 147 - 51 Penetration of topical fluconazole into human aqueous humor; Abbasoglu OE et al.; The purpose of this study was to determine the aqueous levels and pharmacokinetics of topical fluconazole 0.2% upon single and multiple drop applications . Forty-nine patients undergoing cataract surgery were given topical fluconazole 0.2% . They either received single drop or a loading dose of 1 drop per 5 min for 20 min . Aqueous samples were obtained during surgery 5, 15, 30, 45 and 60 min after the last drop . The samples were analysed by high-pressure liquid chromatography to determine aqueous concentrations . After single and loading dose applications peak aqueous levels were achieved at 15 min (3.35 +/- 0.64 and 7.13 +/- 0.79 microg ml(-1), respectively) . Both had a steady decrease in concentration at 30, 45 and 60 min down to 4.06 +/- 0.37 microg ml(-1)with loading dose and undetectable levels with single dose application . Comparing the concentrations with the minimum inhibitory concentrations (MIC) of yeasts determined by the National Committee for Clinical Laboratory Standards showed that concentrations achieved with single dose applications were higher than MICs of Candida albicans and Candida parapsilosis and concentrations achieved after loading dose applications were higher than MICs of C . parapsilosis, C . albicans and Candida tropicalis . We concluded that topical fluconazole 0.2% penetrates into the aqueous humor in concentrations that satisfy MICs of most of the Candida strains . It can be a good alternative to Amphotericin B for treatment of Candida keratitis. J Clin Microbiol, 2001 Feb, 39(2), 525 - 32 Influence of glucose supplementation and inoculum size on growth kinetics and antifungal susceptibility testing of Candida spp; Cuenca-Estrella M et al.; The influences of inoculum size and glucose supplementation on the growth kinetics of 60 Candida spp . clinical isolates (Candida albicans, Candida tropicalis, Candida parapsilosis, Candida glabrata, Candida krusei, and Candida lusitaniae {10 isolates each}) are assessed . The combined influence of growth and reading method (visual or spectrophotometric) on the determination of the MICs of amphotericin B, flucytosine, fluconazole, itraconazole, ketoconazole, and voriconazole is also analyzed, and the MICs are compared with those determined by the National Committee for Clinical Laboratory Standards standard microdilution method (NCCLS document M27-A) . Glucose supplementation and inoculum size had a significant influence on the growth cycles of these yeasts, and a statistically significant denser growth (optical density at 540 nm) was seen for both incubation periods, 24 and 48 h (P < 0.01) . A longer exponential phase and shorter lag phase were also observed . The A540 values at 24 h of incubation with medium containing glucose and an inoculum of 10(5) CFU/ml were >0.4 U for all species, with the exception of that for C . parapsilosis (A540 = 0.26 +/- 0.025) . The MICs at 24 h determined by testing with 2% glucose and an inoculum of 10(5) CFU/ml showed the strongest agreement (96.83%) with MICs determined by the reference method . MICs were not falsely elevated, and good correlation indexes were obtained . The reproducibility of results with this medium-inoculum combination was high (intraclass correlation coefficient, 0.955) . The best agreement and reproducibility of results for spectrophotometric readings were achieved with endpoints of 50% growth inhibition for flucytosine and azoles and 95% for amphotericin B . Supplementation of test media with glucose and an inoculum size of 10(5) CFU/ml yielded a reproducible technique that shows elevated agreement with the reference procedures and a shorter incubation period for obtaining reliable MIC determinations . The spectrophotometric method offers an advantage over the visual method by providing a more objective and automated MIC determination. FEBS Lett, 2000 Dec 22, 487(1), 91 - 4 Genomic exploration of the hemiascomycetous yeasts: 16 . Candida tropicalis; Blandin G et al.; The genome of the diploid hemiascomycetous yeast Candida tropicalis, an opportunistic human pathogen and an important organism for industrial applications, was explored by the analysis of 2541 Random Sequenced Tags (RSTs) covering about 20% of its genome . Comparison of these sequences with Saccharomyces cerevisiae and other species permitted the identification and the analysis of a total of more than 1000 novel genetic elements of C . tropicalis . Moreover, the present study confirms that in C . tropicalis, the rare CUG codon is read as a serine and not a leucine . The sequences have been deposited at EMBL with the accession numbers AL438875-AL441602. Pediatrics, 2001 Jan, 107(1), 61 - 6 Review of 49 neonates with acquired fungal sepsis: further characterization; Makhoul IR et al.; BACKGROUND: Neonatal acquired fungal sepsis (AFS) is a risky condition that warrants every effort for early diagnosis and management . METHODS: We retrospectively reviewed the medical charts of all 4445 neonatal intensive care unit (NICU) admissions in the past 10 years and detected 49 neonates with AFS . We then compared their data with those of 49 matched control neonates who did not have AFS . The following details were collected: gestational, perinatal and neonatal courses; bacterial sepsis; antibacterial therapy; laboratory and imaging investigations; and antifungal therapy and its complications . RESULTS: The incidence of AFS was.4 to 2 cases per 1000 live-births and 3.8% to 12.9% of very low birth weight (VLBW) infants . Compared with 1989 through 1992, between 1993 and 1995 the rate of AFS in VLBW neonates significantly increased (3 . 8%-5.6% --> 9.6%-12.9%), along with a significant increase of NICU admission rate (369-410 --> 496-510 admissions/year) . Compared with controls, AFS neonates had significantly longer hospitalizations, higher rates of mechanical ventilation, umbilical vein catheterization, and previous treatment with broad-spectrum antibacterial agents (amikacin, vancomycin, ceftazidime, or imipenem) . At the onset of AFS, 42.8% of patients had hyperthermia and 40.9% had normal white blood cell count . Causative fungi were as follows: Candida albicans-42.8% of cases, Candida parapsilosis-26.5%, and Candida tropicalis-20.4% . Fungal dissemination was rare, complications of antifungal therapy were infrequent, and no deaths occurred . CONCLUSIONS: First, non-albicans Candida have become more frequent in neonatal AFS . Second, mechanical ventilation and antibacterial agents are significant risk factors for AFS . Third, hyperthermia is a frequent presenting sign of AFS . Fourth, a normal white blood cell count does not rule out AFS . Fifth, meningeal involvement in neonatal AFS should be ruled out before initiation of antifungal therapy . Sixth, the policy of empiric antifungal therapy for AFS should be considered on an individual NICU basis.newborn infant, fungal sepsis, clinical signs, risk factors. J Infect, 2000 Nov, 41(3), 275 - 6 Candida tropicalis pacemaker endocarditis; Kurup A et al.; A rare case of Candida tropicalis pacemaker endocarditis was diagnosed in a 77-year-old male who presented with lethargy . The organism was isolated from cultures of blood and vegetations on the tricuspid valve, interatrial septum and the pacing wire removed at surgery . The postoperative course was stormy and he succumbed to multiorgan failure . Infect Immun, 2001 Jan, 69(1), 405 - 12 Secreted aspartic proteinase family of Candida tropicalis; Zaugg C et al.; Medically important yeasts of the genus Candida secrete aspartic proteinases (Saps), which are of particular interest as virulence factors . Like Candida albicans, Candida tropicalis secretes in vitro one dominant Sap (Sapt1p) in a medium containing bovine serum albumin (BSA) as the sole source of nitrogen . Using the gene SAPT1 as a probe and under low-stringency hybridization conditions, three new closely related gene sequences, SAPT2 to SAPT4, encoding secreted proteinases were cloned from a C . tropicalis lambdaEMBL3 genomic library . All bands identified by Southern blotting of EcoRI-digested C . tropicalis genomic DNA with SAPT1 could be assigned to a specific SAP gene . Therefore, the SAPT gene family of C . tropicalis is likely to contain only four members . Interestingly, the SAPT2 and SAPT3 gene products, Sapt2p and Sapt3p, which have not yet been detected in C . tropicalis cultures in vitro, were produced as active recombinant enzymes with the methylotrophic yeast Pichia pastoris as an expression system . As expected, reverse transcriptase PCR experiments revealed a strong SAPT1 signal with RNA extracted from cells grown in BSA medium . However, a weak signal was obtained with all other SAPT genes under several conditions tested, showing that these SAPT genes could be expressed at a basic level . Together, these experiments suggest that the gene products Sapt2p, Sapt3p, and Sapt4p could be produced under conditions yet to be described in vitro or during infection. Infect Immun, 2001 Jan, 69(1), 108 - 14 Single-copy IMH3 allele is sufficient to confer resistance to mycophenolic acid in Candida albicans and to mediate transformation of clinical Candida species; Beckerman J et al.; Parasexual genetic analysis of Candida albicans utilized the dominant selectable marker that conferred resistance to mycophenolic acid . We cloned and sequenced the IMH3(r) gene from C . albicans strain 1006, which was previously identified as resistant to mycophenolic acid (MPA) (A . K . Goshorn and S . Scherer, Genetics 123:213-218, 1989) . MPA is an inhibitor of IMP dehydrogenase, an enzyme necessary for the de novo biosynthesis of GMP . G . A . Kohler et al . (J . Bacteriol . 179:2331-2338, 1997) have shown that the wild-type IMH3 gene, when expressed in high copy number, will confer resistance to this antibiotic . We demonstrate that the IMH3(r) gene from strain 1006 has three amino acid changes, two of which are nonconservative, and demonstrate that at least two of the three mutations are required to confer resistance to MPA . We used this gene as a dominant selectable marker in clinical isolates of C . albicans and Candida tropicalis . We also identified the presence of autonomously replicating sequence elements that permit autonomous replication in the promoter region of this gene . Finally, we found the excision of a phi-type long terminal repeat element outside the IMH3 open reading frame of the gene in some strains . We used the IMH3(r) allele to disrupt one allele of ARG4 in two clinical isolates, WO-1 and FC18, thus demonstrating that a single ectopic integration of this dominant selectable marker is sufficient to confer resistance to MPA. Braz J Infect Dis, 1998 Aug, 2(4), 187 - 196 Effect of Antifungal Agents on Candida spp . and Pichia anomala Isolated from Oropharyngeal Candidiasis of AIDS Patients in a University Hospital in Brazil; Branchini ML et al.; In order to determine the clinical significance of oropharyngeal candidiasis in AIDS patients, 44 isolates collected from individual clinical episodes were evaluated . The isolates were identified by microbiologic standard methods and in vitro antifungal susceptibility testing was evaluated for amphotericin B, fluconazole, flucytosine, miconazole, itraconazole, and ketoconazole according to the National Committee for Clinical Laboratory Standards . Candida albicans ATCC90028 was used as control for the MICs . The microbiologic isolation revealed 2 strains of Pichia anomala, an uncommon pathogen in AIDS patients . C.albicans and Candida tropicalis comprised 35 and 6 isolates, respectively . The antifungal susceptibility testing for C.albicans isolates (35 isolates) revealed low sensitivity and 19 strains (54%) were resistant to at least 1 antifungal agent . 5 strains (14%) showed multi-drug resistance, including to fluconazole . The resistant profiles of these Candida spp . are of major concern since at present, fluconazole is not commonly prescribed for the treatment of oral candidiasis in our AIDS patients . All C.tropicalis isolates were resistant to fluconazole and miconazole . Both P.anomala strains were resistant to fluconazole . The local diversity of species and the variability of MICs and IC80s of antifungal agents seen in this study indicate that continuing evaluation of clinical and laboratory aspects of oral candidiasis in our HIV1-patients is needed, and that a new approach and therapy will be necessary to manage the increasing problem. Steroids, 2001 Jan, 66(1), 49 - 54 Expression of 17beta-hydroxysteroid dehydrogenases in mesophilic and extremophilic yeast; Rizner TL et al.; 17beta-hydroxysteroid dehydrogenases (17beta-HSDs) are enzymes responsible for reversible interconversions of biologically active 17-hydroxy and inactive 17-keto steroids . We have performed a survey of 17beta-HSD activity in yeast . Constitutive 17beta-HSD activity was found in three mesophilic yeast species: Candida tropicalis, Cryptococcus tsukubaensis, and Saccharomyces cerevisiae as well as in three extremophilic black yeast species: Hortaea werneckii, Trimmatostroma salinum, and Phaeotheca triangularis, indicating that 17beta-HSD activity is widely distributed among yeast . In extremophilic black yeast, NaCl modulated enzyme activity . Enzymes resembling 17beta-HSD from the filamentous fungus Cochliobolus lunatus were detected in Trimmatostroma salinum and Phaeotheca triangularis . Sequences with identity to the Saccharomyces cerevisiae YBR159w gene were not observed in other yeast species possessing a similar enzyme activity . The results suggest the existence of at least three different types of 17beta-HSD in yeast. Eur J Obstet Gynecol Reprod Biol, 2000 Dec, 93(2), 141 - 5 Determinants of different Candida species infections of the genital tract in women . Sporachrom Study Geoup; Parazzini F et al.; OBJECTIVE: We have analyzed the differences in the epidemiological characteristics of women with different Candida low female genital tract infection . STUDY DESIGN: Eligible for the study were 4228 women aged 18-70 years with symptomatic low gynecological tract infection and clinical findings suggestive for Candida infection consecutively attending during the study period first level outpatients gynecological services in Italy . CHROMagar Candida method was used to identify albicans and non-albicans species and among non-albicans ones Candida glabrata, tropicalis and krusei . RESULTS: Out of the 4228 women who entered the study, Candida infection was confirmed by CHROMagar test in 3351 cases (79.3%): Candida albicans was identified in 1431 cases (43%) and non-albicans in 1920 . Among the 1920 women with non-albicans infection, Candida glabrata was identified in 1207 women, Candida krusei in 290, Candida tropicalis in 404 (in 19 cases other species or non-specified species were involved) . Candida albicans infection was more frequently reported than non-albicans ones in diabetic women (Odds Ratio, OR=1.7, 95%, Confidence Interval, CI 1.1-2.7) . Current oral contraceptive users tended more frequently to be infected with Candida albicans than non-albicans, however the estimated OR was only slightly above unity and of borderline statistical significance (OR 1.3, 9.5%, CI 1.1-1.5) . Women reporting previous treatment with topic antimicotic reported more frequently non-albicans infection, than Candida albicans ones . However the association was limited and of borderline statistical significance (OR albicans vs . non albicans 0.7, 95% CI 0.5-1.0) . Albicans infection was more frequently identified in women whose partner reported symptomatology for Candida infection (OR 1.7, 95% C.I . 1.4-2.0) . CONCLUSIONS: This study shows that in this Italian population with symptomatic Candida infection of low female genital tract, there are some differences in the epidemiological characteristics of women with albicans and non-albicans infection. Arch Med Res, 2000 Jul-Aug, 31(4), 388 - 92 Microbiological findings in febrile neutropenia; Gaytan-Martinez J et al.; BACKGROUND: This study was carried out to assess the isolation rate of bacterial and fungal causative agents in Mexican neutropenic adults with hematological neoplasia . METHODS: A prospective observational survey involving 120 consecutive episodes of febrile neutropenia during 1 year was carried out . These episodes were observed in 630 patients discharged with diagnoses of leukemia or lymphoma, or after bone-marrow transplantation . RESULTS: At least one pathogen was isolated in 42 of 120 episodes (35%), and was present in 39 patients with acute myeloid leukemia (AML) (43%), acute lymphoblastic leukemia (ALL) (23%), and in patients who underwent bone-marrow transplantation (20%) . Primary bacteremia was the most frequent cause of fever (24 episodes, 57%), followed by intravascular device-related infections (5 episodes, 17%), and soft-tissue infections (5 episodes, 15%) . Escherichia coli (33%) was the most frequently isolated agent of primary bacteremia, followed by coagulase-negative Staphylococcus (29%), and Klebsiella oxytoca (16%) . Fungal infection was responsible for five events (4%): two episodes of pneumonia (Penicillium marneffei and Aspergillus fumigatus, one event each); two cases of fungemia, one due to Candida tropicalis and one to Rhodotorula gluttinis, and one cryptococcal meningitis event . CONCLUSIONS: The isolation rate, approximately 30%, was in accordance with previous reports; similar percentages of Gram-positive and Gram-negative isolates were found . A remarkably low rate of viridans group streptococci and fungal agents was observed, despite the fact that neutropenia is the main risk factor for infection due to these agents . Studies reporting local microbiological findings are necessary because they support an antibiotic choice for prophylaxis or therapy more accurately than reports from other areas. Expert Opin Investig Drugs, 2000 Mar, 9(3), 593 - 605 Novel triazole antifungal agents; Hoffman HL et al.; The risk of opportunistic infections is greatly increased in patients who are immunocompromised due to AIDS, cancer chemotherapy and organ or bone marrow transplantation . Candida albicans is often associated with serious systemic fungal infections, however other Candida species such as Candida krusei, Candida tropicalis and Candida glabrata, as well as Cryptococcus neoformans and filamentous fungi such as Aspergillus, have also emerged as clinically significant fungal pathogens . Two triazole antifungal agents, fluconazole and itraconazole, were introduced over a decade ago and since then have been used extensively for the prophylaxis and treatment of a variety of fungal infections . Although both drugs are effective and have their place in therapy, limitations regarding the utility of these agents do exist . For example, fluconazole is not effective for the prophylaxis or treatment of Aspergillus species and has limited activity against C . krusei and C . glabrata . The use of itraconazole has been limited secondary to concerns regarding unpredictable bioavailability . The rising incidence of fungal infections and the reported increase of non-albicans candidal infections noted over the past two decades highlight the need for new antifungal agents with improved spectra of activity . Several new triazole agents are in various phases of preclinical and clinical trials and may be available for human use in the near future . Three such agents voriconazole, posaconazole and ravuconazole are reviewed and compared with existing agents. J Clin Microbiol, 2000 Nov, 38(11), 4186 - 92 Panfungal PCR and multiplex liquid hybridization for detection of fungi in tissue specimens; Hendolin PH et al.; A procedure based on panfungal PCR and multiplex liquid hybridization was developed for the detection of fungi in tissue specimens . The PCR amplified the fungal internal transcribed spacer (ITS) region (ITS1-5.8S rRNA-ITS2) . After capture with specific probes, eight common fungal pathogens (Aspergillus flavus, Aspergillus fumigatus, Candida albicans, Candida krusei, Candida glabrata, Candida parapsilosis, Candida tropicalis, and Cryptococcus neoformans) were identified according to the size of the amplification product on an automated sequencer . The nonhybridized products were identified by sequencing . The performance of the procedure was examined with 12 deep-tissue specimens and 8 polypous tissue biopsies from the paranasal sinuses . A detection level of 0.1 to 1 pg of purified DNA (2 to 20 CFU) was achieved . Of the 20 specimens, PCR was positive for 19 (95%), of which 10 (53%) were hybridization positive . In comparison, 12 (60%) of the specimens were positive by direct microscopy, but only 7 (35%) of the specimens showed fungal growth . Sequencing of the nonhybridized amplification products identified an infecting agent in six specimens, and three specimens yielded only sequences of unknown fungal origin . The procedure provides a rapid (within 2 days) detection of common fungal pathogens in tissue specimens, and it is highly versatile for the identification of other fungal pathogens. Adv Perit Dial, 2000, 16, 233 - 6 Role of intraperitoneal urokinase in acute peritonitis and prevention of catheter loss in peritoneal dialysis patients; Gadallah MF et al.; Some studies have demonstrated the efficacy and safety of intraperitoneal (i.p.) urokinase in the resolution of recurrent or relapsing peritonitis, while others have not . Most studies were small, and they varied in methodology . Furthermore, the role of i.p . urokinase in shortening the duration of peritonitis or in preventing recurrence after initial peritonitis has not been examined . In addition, no previous studies have examined the role of i.p . urokinase in preventing, after first infection, catheter loss due to unresolving (resistant) peritonitis . Over a period of 3 years, we prospectively randomized into two groups all peritoneal dialysis (PD) patients who developed a first episode of peritonitis . Group I (n = 40) received i.p . urokinase on the first day of diagnosis (5000 IU intraluminally in the peritoneal catheter and left for 4 hours before next exchange) . Group II (n = 40) received no urokinase . The duration of peritonitis was assessed by daily PD fluid white blood cell (WBC) count . Indications for catheter removal were: persistent peritonitis after four days from initiation of antibiotic therapy, or peritonitis with multiple organisms, suggesting bowel perforation . No statistically significant difference was seen between the two groups in regard to primary cause of end-stage renal disease (ESRD), age, sex, race, weight, type of dialysis {continuous ambulatory peritoneal dialysis (CAPD), automated peritoneal dialysis (APD), continuous cycling peritoneal dialysis (CCPD)}, or duration of dialysis prior to first peritonitis . No statistically significant difference was seen between the two groups in the duration of peritonitis or in the severity of symptoms and signs of peritonitis . Neither was any difference seen in the peritonitis recurrence or relapse rate (10% in the urokinase group vs 7.5% in the control group) . Nine patients lost their catheters (3 in the urokinase group: 1 Pseudomonas aeruginosa and 2 Candida tropicalis; 6 in the control group: 1 Klebsiella pneumonia, 1 enterococcus, 2 Pseudomonas aeruginosa, and 2 Candida tropicalis) . The difference in the rate of catheter loss between the two groups was not statistically significant; it appeared to relate to the type of organism rather than to the response to urokinase . In conclusion, i.p . urokinase plays no significant role in shortening the course of peritonitis or in preventing recurrence or loss of the PD catheter . Loss of PD catheters in patients having their first peritonitis appears to be related primarily to the type of organism causing the infection. Diagn Microbiol Infect Dis, 2000 Sep, 38(1), 21 - 8 Sustained gastrointestinal colonization and systemic dissemination by Candida albicans, Candida tropicalis and Candida parapsilosis in adult mice; Mellado E et al.; The ability of nine clinical isolates of Candida species (three C . albicans, three C . tropicalis and three C . parapsilosis) to colonize and invade the gastrointestinal (GI) tract of adult male CD-1 (ICR) mice was determined . The effect of dietary tetracycline plus glucose supplementation on colonization was evaluated . Strains were intragastrically inoculated . Tetracycline and glucose altered substantially aerobic flora, especially streptococci (average fall 1.1 +/-0.3 log(10) CFU/g, p<0.01 by the Student's t test) . At two weeks after oral challenge, sustained and high colonization of GI tract by Candida (mean 5,28 +/- 0.18 log(10) CFU/g, p<0.01) was achieved in all mice receiving glucose-tetracycline supplementation, excepting in animals inoculated with one of C . tropicalis isolates . Histologic sections of the stomachs revealed multiple intraepithelial micro-abscesses associated with hyphae in the region of the cardial-atrium fold . Under immunosuppression, systemic spread of C . albicans and C . tropicalis was observed in 62% and 24% of animals receiving dietary supplementation respectively . Dissemination was not noted for C . parapsilosis isolates . We have developed a simple and inexpensive murine model of sustained colonization of GI tract . This model could be useful for analyzing prophylaxis, treatment and diagnosis of systemic Candida infections and for evaluating virulence of strains. J Clin Microbiol, 2000 Oct, 38(10), 3735 - 42 Development of a PCR-based line probe assay for identification of fungal pathogens; Martin C et al.; We report on a reverse-hybridization line probe assay (LiPA) which when combined with PCR amplification detects and identifies clinically significant fungal pathogens including Candida, Aspergillus, and Cryptococcus species . DNA probes have been designed from the internal transcribed-spacer (ITS) regions of Candida albicans, Candida parapsilosis, Candida glabrata, Candida tropicalis, Candida krusei, Candida dubliniensis, Cryptococcus neoformans, Aspergillus fumigatus, Aspergillus versicolor, Aspergillus nidulans and Aspergillus flavus . The probes were incorporated into a LiPA for detection of biotinylated ITS PCR products, and the specificity of the probes was evaluated . We established LiPA detection limits for ITS 1 and for full ITS amplicons for genomic DNA from C . albicans, A . fumigatus, and C . neoformans . Further evaluation of the LiPA was carried out on clinical fungal isolates . One hundred twenty-seven isolates consisting of dimorphic yeasts and dermatophytic and filamentous fungi were tested by the LiPA, which correctly identified 77 dimorphic yeasts and 23 of the filamentous isolates; the remaining 27 isolates represented species of fungi for which probes were not included in the LiPA . The fungal-PCR-LiPA technology was applied to blood samples inoculated with Candida cells which were pretreated by minibead beating to mechanically disrupt the cells, with the DNA extracted by either a previously described guanidium thiocyanate-silica method or the commercially available QIAmp tissue kit . PCR amplification of the extracted DNA and subsequent DNA probe hybridization in the LiPA assay yielded detection limits of 2 to 10 cells/ml . An internal standard control was included in the PCR amplification to monitor for PCR inhibition . This fungal PCR-LiPA assay is robust and sensitive and can easily be integrated into a clinical-testing laboratory with the potential for same-day diagnosis of fungal infection. Eur J Clin Microbiol Infect Dis, 2000 Aug, 19(8), 602 - 7 Nosocomial candidemia in non-neutropenic patients at an Italian tertiary care hospital; Luzzati R et al.; In a retrospective study conducted in an Italian tertiary care hospital, the incidence of nosocomial candidemia was evaluated together with causative pathogens, treatment, and risk factors for death . Over a 6-year period (1992-1997), a total of 189 episodes of candidemia occurred in 189 patients (mean age 58+/-19 years), accounting for an average incidence of 1.14 episodes per 10,000 patient-days per year . The most common reasons for hospitalization were solid neoplasia (21%), trauma (17%), abdominal diseases requiring surgery (13%), and cardiovascular diseases (13%) . No patient was neutropenic within 3 weeks prior to the onset of candidemia . One hundred thirty patients were hospitalized in intensive care units, 47 patients in surgical wards, and 12 patients in medical wards . Candida albicans was the most frequently isolated pathogen, accounting for 54% of fungal isolates, followed by Candida parapsilosis (23%), Candida glabrata (7%), Candida tropicalis (5%), Candida pelliculosa (4%), Candida lusitaniae (1%), Candida humicula (1%), and other non-albicans Candida spp . (5%) . Seventy-six (41%) patients received adequate antifungal therapy . Seventy-one (58%) of the 123 evaluable patients with central venous catheters underwent line removal; 51 of them had catheter-related candidemia . The 30-day crude mortality rate was 45% . Older age, hospitalization in an intensive care unit, a longer duration of candidemia, retention of central lines, and inadequate antifungal therapy were significantly associated with poor outcome . In the present study, nosocomial candidemia was a frequent and relatively underestimated illness . Adequate antifungal therapy and central line removal independently reduced the high mortality of the disease. Antimicrob Agents Chemother, 2000 Oct, 44(10), 2752 - 8 Flow cytometry antifungal susceptibility testing of pathogenic yeasts other than Candida albicans and comparison with the NCCLS broth microdilution test; Ramani R et al.; Candida species other than Candida albicans frequently cause nosocomial infections in immunocompromised patients . Some of these pathogens have either variable susceptibility patterns or intrinsic resistance against common azoles . The availability of a rapid and reproducible susceptibility-testing method is likely to help in the selection of an appropriate regimen for therapy . A flow cytometry (FC) method was used in the present study for susceptibility testing of Candida glabrata, Candida guilliermondii, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, and Cryptococcus neoformans based on accumulation of the DNA binding dye propidium iodide (PI) . The results were compared with MIC results obtained for amphotericin B and fluconazole using the NCCLS broth microdilution method (M27-A) . For FC, the yeast inoculum was prepared spectrophotometrically, the drugs were diluted in either RPMI 1640 or yeast nitrogen base containing 1% dextrose, and yeast samples and drug dilutions were incubated with amphotericin B and fluconazole, respectively, for 4 to 6 h . Sodium deoxycholate and PI were added at the end of incubation, and fluorescence was measured with a FACScan flow cytometer (Becton Dickinson) . The lowest drug concentration that showed a 50% increase in mean channel fluorescence compared to that of the growth control was designated the MIC . All tests were repeated once . The MICs obtained by FC for all yeast isolates except C . lusitaniae were in very good agreement (within 1 dilution) of the results of the NCCLS broth microdilution method . Paired t test values were not statistically significant (P = 0.377 for amphotericin B; P = 0.383 for fluconazole) . Exceptionally, C . lusitaniae isolates showed higher MICs (2 dilutions or more) than in the corresponding NCCLS broth microdilution method for amphotericin B . Overall, FC antifungal susceptibility testing provided rapid, reproducible results that were statistically comparable to those obtained with the NCCLS method. Antimicrob Agents Chemother, 2000 Oct, 44(10), 2664 - 71 In vivo activity of amphotericin B lipid complex in immunocompromised mice against fluconazole-resistant or fluconazole-susceptible Candida tropicalis; Warn PA et al.; We compared four doses of amphotericin B lipid complex (ABLC) with three doses of fluconazole in temporarily neutropenic mice in a murine model of disseminated candidiasis due to four different isolates of Candida tropicalis . The mice were infected with a 90% lethal dose of four strains of C . tropicalis for which the fluconazole MICs ranged from 1 to >125 mg/liter 3 days after receiving 200 mg of cyclophosphamide/kg of body weight . Treatment was started 18 h after infection and lasted for 7 days . ABLC (1, 2, 5, and 10 mg/kg) was administered once a day intravenously, fluconazole was administered by oral gavage once daily (25 and 50 mg/kg/day) or twice daily (125 mg/kg) . MICs determined in five different ways with 24- and 48-h endpoints were also compared . The overall survival rates were controls, 14%; fluconazole, 64%; and ABLC, 82% . Treatment with ABLC at 2 to 10 mg/kg increased survival compared to controls (P = <0.0001) and was also superior to fluconazole at 25 and 50 mg/kg (P = 0.006) . In the fluconazole-resistant C . tropicalis model (MIC, 128 microg/ml), ABLC at 2 to 10 mg/kg was superior to fluconazole at 250 mg/kg and ABLC at 10 mg/kg was superior to all fluconazole doses (P = <0.05) . Fluconazole at 250 mg/kg daily was superior to both 25 and 50 mg/kg at reducing mortality with most isolates . ABLC was superior to fluconazole (P = <0.01), and fluconazole at 250 mg/kg was superior to fluconazole at both 25 and 50 mg/kg (P = 0.02) in all models at reducing C . tropicalis counts in the kidneys . Neither drug consistently sterilized the brain or kidneys . A 48-h endpoint reading with the NCCLS susceptibility testing microtiter variation overestimates resistance to fluconazole . ABLC is an effective treatment for fluconazole-resistant C . tropicalis at all doses tested. Clin Infect Dis, 2000 Aug, 31(2), 451 - 7 Epub 2000 Aug 28. Candidal meningitis in children with cancer; McCullers JA et al.; Candidal meningitis is a rare disease that is seen most frequently in neonates, neurosurgical patients, and the immunocompromised host . We describe a series of 12 children with cancer (all of whom had leukemia) who had candidal meningitis develop . Univariate analysis revealed that duration of fever, antibiotic therapy, and profound neutropenia and use of total parenteral nutrition were significantly associated (P<.05) with candidal meningitis in children with cancer, compared with matched control subjects . Only duration of profound neutropenia (P=.08) and use of total parenteral nutrition (P=.06) approached significance in the multivariate analysis . One species of Candida, Candida tropicalis, was responsible for 11 of the 12 cases, indicating increased pathogenicity of this organism in CNS disease . The cases were invariably fatal, supporting aggressive treatment of candidal meningitis in immunocompromised patients and further study of the prevention, diagnosis, and management of C . tropicalis meningitis. J Antimicrob Chemother, 2000 Sep, 46(3), 485 - 7 In vitro antifungal activity of FK463, a new water-soluble echinocandin-like lipopeptide; Mikamo H et al.; The antifungal activity of FK463 against 72 recent clinical isolates of Candida albicans (24), Candida glabrata (17), Candida tropicalis (11), Candida krusei (8) and Candida parapsilosis (12) was compared with those of amphotericin B, fluconazole and itraconazole by means of a broth microdilution method specified by the National Committee for Clinical Laboratory Standards (NCCLS) document M27-A . The lowest drug concentration at which 90% of the population was inhibited (MIC(90)) of FK463 against C . albicans, C . glabrata, C . tropicalis, C . krusei and C . parapsilosis was 0.0156, 0 . 0156, 0.0313, 0.125 and 1 mg/L, respectively . FK463 exhibited broad-spectrum activity against clinically important Candida spp . (MIC range < or =0.0039-2 mg/L), and its MICs for such fungi were lower than those of other antifungal agents tested . The minimum fungicidal concentrations for Candida spp . did not differ by more than two-fold from the MICs . Results from pre-clinical evaluations performed to date indicate that FK463 should be a potent parenteral antifungal agent. J Antimicrob Chemother, 2000 Sep, 46(3), 475 - 7 Susceptibility of fluconazole-resistant clinical isolates of Candida spp . to echinocandin LY303366, itraconazole and amphotericin B; Cuenca-Estrella M et al.; The in vitro activity of LY303366 was compared with those of itraconazole and amphotericin B against 156 fluconazole-resistant (MIC > or = 16 mg/L) clinical isolates of CANDIDA: spp . An adaptation of the NCCLS reference method was employed for determination of MICs . LY303366 was more potent than either itraconazole or amphotericin B against Candida albicans, Candida glabrata, Candida krusei and Candida tropicalis, even against isolates with itraconazole MICs > or = 1 mg/L . LY303366 was less potent in vitro against Candida parapsilosis and Candida guilliermondii isolates . LY303366 has promising antifungal activity and warrants further investigation. J Infect Dis, 2000 Oct, 182(4), 1276 - 9 Epub 2000 Sep 06. Differential host susceptibility to pulmonary infections with bacteria and fungi in mice deficient in myeloperoxidase; Aratani Y et al.; Myeloperoxidase (MPO), which is located within neutrophils capable of producing hypochlorous acid, is active in vitro against bacteria and fungi . However, MPO-deficient persons are usually healthy . To define the in vivo contribution of MPO to early host defense against pulmonary infections, MPO-deficient and control mice were intranasally infected with various fungi and bacteria, and the number of residual microorganisms in lungs was compared 48 h later . MPO-deficient mice showed severely reduced cytotoxicity to Candida albicans, Candida tropicalis, Trichosporon asahii, and Pseudomonas aeruginosa . However, the mutant mice showed a slight but significantly delayed clearance of Aspergillus fumigatus and Klebsiella pneumoniae and had comparable levels of resistance to the wild type against Candida glabrata, Cryptococcus neoformans, Staphylococcus aureus, and Streptococcus pneumoniae . These results suggest that the MPO-dependent oxidative system is important for host defense against fungi and bacteria, although the effect varies by pathogen species. Sheng Wu Gong Cheng Xue Bao, 2000 Mar, 16(2), 198 - 202 {Studies on fermentation of decane 1,10-dicarboxylic acid(DC12)}; Ren G et al.; Candida tropicalis strain UH-2-48 can increase the production of decane 1,10-dicarboxylic acid(DC12) yield by 30% (from 81.7 g/L to 108.2 g/L) after regulating the fermentation condition . The purity of the DC12 is above 97%, which can meet the criteria of the succeeding synthesized industrial productions . Emulsifier can not only destroy the structure of the cell membrane in some degree, but also can enhance the permeability of the cell membrane . Furthermore, the structure of the emulsifier itself can also effect its function . Tween 60(0.10%) can enhance the production of DC12 tremendously . Urea, as the nitrogen resource, can affect the activity of cytochrome P450 enzyme . Some biochemistry and biophysics factors, such as penicillin, vitamin B2 and alanine, can increase the yield of DC12 during the process of fermentation . Ferment in 20 m3 fermentator, under the optimal condition, the average yield of DC12 using the strain HP-12 which derived from UH-2-48, is 202.1 g/L. FEMS Immunol Med Microbiol, 2000 Sep, 29(1), 35 - 8 A PCR-based approach to sequence the Candida tropicalis HSP90 gene; Santhanam J et al.; The heat shock protein 90 (hsp90) gene sequence is known to be highly conserved across the species barrier . A PCR-based method was thus utilised in an attempt to sequence the Candida tropicalis hsp90 gene . Primers for PCR were designed from conserved regions of the gene, which were identified by comparing the Saccharomyces cerevisiae and Candida albicans hsp90 gene sequences . Different sets of primers were designed to amplify and obtain overlapping DNA sequences of the C tropicalis gene . PCR was carried out on genomic DNA of Candidca tropicalis and the PCR products were cloned into suitable vector molecules for sequencing . In this way, a 2,070-basepair sequence of the C . tropicalis hsp90 gene was obtained . The PCR-based approach proved to be an easier method of obtain the sequence of a highly conserved gene, as compared to more conventional methods. Infection, 2000 Jul-Aug, 28(4), 223 - 6 Liposomal amphotericin B (AmBisome) in the treatment of neonatal candidiasis in very low birth weight infants; Juster-Reicher A et al.; AmBisome (2.5-7 mg/kg/day as a continuous 1 h infusion) was evaluated prospectively from September 1994 to January 1998 in 24 very low birth weight infants (mean birth weight 847+/-244 g, mean gestational age 26 weeks) with systemic candidiasis . Mean age at onset of candidemia was 17 days . One patient had two episodes of candidiasis.Thirteen infants failed previous antifungal therapy with amphotericin B (with or without 5-flucytosine) . Candida spp . were isolated from the blood in all 25 episodes and from skin abscesses and urine in four infants each, respectively . There were 13 isolates of Candida albicans, ten of Candida parapsilosis, two of Candida tropicalis and one of Candida glabrata . One infant had a mixed infection with C . albicans and C . parapsilosis . The mean duration of therapy was 21 days; the cumulative AmBisome dose was 94 mg/kg . Fungal eradication was achieved in 92% of the episodes; mean duration of AmBisome therapy until achieving eradication was 9 days . Twenty (83%) infants were considered clinically cured at the end of treatment . No major adverse effects were recorded; one infant developed increased bilirubin and hepatic transaminases levels during therapy . Four (17%) infants died; in two of them (8%) the cause of death was directly attributed to systemic candidiasis . CONCLUSION: AmBisome represents an effective, safe and convenient antifungal agent in the therapy of systemic fungal infections in very low birth weight infants. J Basic Microbiol, 2000, 40(3), 167 - 75 Effect of D-glucose on induction of xylose reductase and xylitol dehydrogenase in Candida tropicalis in the presence of NaCl; Ikeuchi T et al.; Xylitol production is suppressed by D-glucose . We previously reported that the suppression was abrogated in the presence of NaCl . This was explained in part by high levels of xylose reductase (XR) activity . In this study, we investigated the effects of D-glucose on XR and xylitol dehydrogenase (XDH) induction in Candida tropicalis in the presence of NaCl . We examined the time courses of these activities under the following conditions: 50 mg/ml D-xylose, 20-80 mg/ml D-glucose and 40 mg/ml NaCl . The level of XR increased in the presence of 40 mg/ml NaCl, whereas that of XDH was not affected by NaCl . The effects of NaCl upon XR and XDH induction suggest that the synthesis of both enzymes is not under a coordinate control . The expression of XR is suppressed by D-glucose . In the absence of NaCl, suppression continued even after D-glucose was completely consumed, while in the presence of 40 mg/ml NaCl, suppression stopped after the consumption of D-glucose. Appl Microbiol Biotechnol, 2000 Jul, 54(1), 126 - 32 Competition between n-alkane-assimilating yeasts and bacteria during colonization of sandy soil microcosms; Schmitz C et al.; An n-alkane-assimilating strain of Candida tropicalis was selected in sandy soil inoculated with microorganisms from contaminated sites . Competition experiments with n-alkane utilizers from different strain collections confirmed that yeasts overgrow bacteria in sandy soil . Acidification of the soil is one of the colonization factors useful for the yeasts . It can be counteracted by addition of bentonite, a clay mineral with high ion exchange capacity, but not, however, by kaolin . Strains of different yeast species showed different levels of competitiveness . Strains of Arxula adeninivorans, Candida maltosa, and Yarrowia lipolytica overgrew strains of C . tropicalis, C . shehatae or Pichia stipitis . Two strains of C . maltosa and Y . lipolytica coexisted during several serial transfers under microcosm conditions. J Chemother, 2000 Aug, 12(4), 339 - 44 Candida meningitis in children: report of two cases; Aldress K et al.; Candida meningitis is rare in children . However reports have been increasing recently . We report two cases of meningitis caused by Candida species . The first case was a term male infant who was admitted at 14 days of age with the diagnosis of possible sepsis . He had received multiple courses of antibiotics without improvement . Later his cerebrospinal fluid (CSF) culture grew Candida tropicalis . The damage done by the infection was severe and the patient died . The second case was a 2-month old girl who was born at 34 weeks of gestation . She was admitted to the Neonatal Intensive Care Unit (NICU) and given antibiotics as prophylaxis . Despite this she developed recurrent episodes of fever that required multiple courses of antibiotics . After discharging her, she continued to have fever . Upon investigation, her blood and CSF grew Candida albicans . She was treated and responded to therapy. New Microbiol, 2000 Jan, 23(1), 73 - 8 Antifungal activity of rhodium, iridium, and ruthenium tripodal phosphine complexes; Sulu M et al.; Twenty-eight rhodium, iridium or ruthenium complexes were evaluated for their in vitro antifungal activities against Candida albicans and Candida tropicalis . Fourteen compounds showed an antifungal activity against C . albicans and C . tropicalis with a range of the minimum inhibitor concentrations (MICs) between 16 and 250 micrograms/mL. New Microbiol, 2000 Jan, 23(1), 63 - 71 Effect of dietary carbohydrates on the in vitro epithelial adhesion of Candida albicans, Candida tropicalis, and Candida krusei; Pizzo G et al.; Adhesion to epithelial surfaces is considered as a critical step in the pathogenesis of oral candidosis . Therefore, the effects of the most commonly consumed dietary carbohydrates on the adhesion of Candida albicans, Candida tropicalis, and Candida krusei to monolayered HeLa cells were investigated . Adherence of C . albicans and C . tropicalis appeared significantly promoted by incubation in defined medium containing a high concentration (500 mM) of fructose, glucose, maltose, and sucrose (p < 0.001) . C . albicans organisms grown in sucrose elicited maximal increase in adhesion, whereas adhesion of C . tropicalis and C . krusei was enhanced to the greatest extent when cultured in glucose . Maltose and fructose also promoted adherence of C . albicans and C . tropicalis (p < 0.001), but to a lesser extent than sucrose and glucose . On the other hand, sorbitol-grown yeasts demonstrated a marginal increase in adhesion (p > 0.01) . Xylitol only significantly reduced adherence of C . albicans (p < 0.001) . These results suggest that the frequent consumption of carbohydrates, such as sucrose, glucose, maltose, or fructose, might represent a risk factor for oral candidosis . The limitation of their consumption by substituting xylitol or sorbitol could be of value in the control of oral Candida colonization and infection. Rev Latinoam Microbiol, 1998 Jan-Jun, 40(1-2), 15 - 24 Fungal agents isolated from cancer patients; Alvarez Gasca MA et al.; With the aim to know the frequency of mycotic agents in patients with different types of cancer, samples were obtained from 81 patients from the Hospital de Oncologia, Centro Medico Nacional Siglo XXI, IMSS from May 1995 through May 1996 . In a conventional grouping seven (7) ambulatory patients were found in early stages, twenty seven (27) occasionally hospitalized patients were found in intermediate stage and forty seven (47) hospitalized patients in terminal stage of cancer . The different samples were processed through routine mycologycal methods and the following fungi species were isolated and identified: fifty four strains (58%) of Candida albicans followed by eleven strains (11.8%) of Candida tropicalis, six strains (6.45%) of Candida parapsilosis, five strains (5.37%) of Candida krusei, four strains (4.3%) of Candida humicola and five strains (5.37%) of Rodothorula rubra . From medical devices like catheter tips, drainage catheters (Pen rouse, Foley) and gallbladder catheters; four (4) strains of C . albicans, three (3) strains of Rodothorula rubra and two (2) strains of Aspergillus sp were isolated . Of the Candida non albicans it was relevant to find C . krusei more frequently than Rodothorula rubra, Aspergillus sp and Penicillum sp . The frequency of the presence of fungi increases commensurately to the advancement of the clincal stage of the cancer. J Hosp Infect, 2000 May, 45(1), 69 - 72 Nosocomial fungaemia: a 2-year prospective study; Costa SF et al.; Eighty-six consecutive patients with fungaemia were studied during a period of 2 years, 81% had two or more positive blood cultures . Gastrointestinal tract (28%) and haematological diseases (17%) were the most common underlying conditions . The majority of cases had received vancomycin and/or imipenem (87%) and a central venous catheter (78%) . Candida albicans (50%) and Candida parapsilosis (17%) were the most frequent isolates . Overall mortality was 41%, and for patients with Candida tropicalis was 71% . There was not significant difference in survival with gender, age and days of treatment with antifungal drugs . Haematological diseases, neutropenia and a higher number of positive blood cultures were associated with poor outcome. Yeast, 2000 Aug, 16(11), 995 - 1000 Establishment of a simple system to analyse the molecular interaction in the agglutination of Saccharomyces cerevisiae; Zou W et al.; Saccharomyces cerevisiae a-agglutinin, which is involved in mating and covalently anchoring to the cell wall, consists of two components, Aga1p and Aga2p, whose syntheses are individually regulated . To facilitate the analysis of the protein-protein interaction on agglutination between a- and alpha-agglutinins, the construction of a yeast strain (MATa) with the functional protein prepared by genetic fusion of Aga1p- and Aga2p-encoding genes and by the expression system using the UPR-ICL promoter derived from the n-alkane-assimilating yeast, Candida tropicalis, which is functional under the condition of lower glucose concentration was tried and the agglutination ability of the constructed strain was evaluated with a yeast strain (MATa) which expressed AGalpha1 encoding alpha-agglutinin under the control of the same promoter . The genes were integrated into the yeast chromosomes . Cell agglutination between both (MATa) strains was observed microscopically when these two strains were mix-cultured to a glucose-decreased concentration . The agglutination was further confirmed by the sedimentation test and by the quantification using a filter . These results proved that the constructed Aga1p-Aga2p fusion protein was enoughly functional for the interaction with the Agalpha1 protein, and that this phenomenon occurred dependent on glucose concentration, but independent of the peptide pheromones secreted by the cells of the opposite mating types . Using this system, the role of two disulphide linkages between Aga1p and Aga2p on the binding activity between Aga2p and Aga1p was first evaluated . Under the treatment by the SH-compound (dithiothreitol), in which Agalpha2p is easily released into the medium from the intact cell surface, the Aga1p and Aga2p fusion protein was a good tool to make clear the role of the disulphide linkages . As a result, the linkages had a significant effect on not only the assembly but also the binding activity . The novel and simple system described here may further facilitate the study of molecular interaction in agglutination . Planta, 2000 Jun, 211(1), 150 - 7 Importance of sequences adjacent to the terminal tripeptide in the import of a peroxisomal Candida tropicalis protein in plant peroxisomes; Bongcam V et al.; The peroxisome targeting signal (PTS) required for import of the rat acyl-CoA oxidase (AOX; EC 1.3.3.6) and the Candida tropicalis multifunctional protein (MFP) in plant peroxisomes was assessed in transgenic Arabidopsis thaliana (L.) Heynh . The native rat AOX accumulated in peroxisomes in A . thaliana cotyledons and targeting was dependent on the presence of the C-terminal tripeptide S-K-L . In contrast, the native C . tropicalis MFP, containing the consensus PTS sequence A-K-I was not targeted to plant peroxisomes . Modification of the carboxy terminus to the S-K-L tripeptide also failed to deliver the MFP to peroxisomes while addition of the last 34 amino acids of the Brassica napus isocitrate lyase, containing the terminal tripeptide S-R-M, enabled import of the fusion protein into peroxisomes . These results underline the influence of the amino acids adjacent to the terminal tripeptide of the C . tropicalis MFP on peroxisomal targeting, even in the context of a protein having a consensus PTS sequence S-K-L. Mycoses, 2000, 43(3-4), 101 - 7 Differentiation of Candida strains by lectin-mediated agglutination kinetics; Nenoff P et al.; The lectin-mediated agglutination kinetics of Candida albicans, Candida tropicalis, Candida glabrata, Candida krusei, Candida kefyr, and Candida parapsilosis strains isolated from immunocompromised patients was investigated . The rate of the lectin-induced cell agglutination depends on the physiological state of the yeast cell population . Therefore, the Candida strains have to be cultivated and investigated under identical conditions . Lentil lectin (prepared from Lens culinaris), castor lectin, and concanavalin A were used . Different yeast species showed different agglutination behaviour . Furthermore, the lectin-mediated rate of agglutination is a strain-specific property which makes it possible to distinguish between different yeast strains of the same species . It is concluded that the lectin-mediated agglutination kinetics allows reproducible differentiation of yeast strains of the same species. Oral Microbiol Immunol, 1999 Dec, 14(6), 358 - 63 Adhesion of oral Candida species to human buccal epithelial cells following brief exposure to nystatin; Ellepola AN et al.; Opportunistic oral infections caused by Candida albicans and non-albicans Candida species are particularly common in compromised patients . Nystatin, which belongs to the polyene group of antimycotics, is frequently used as a topical agent in the treatment of oro-pharyngeal candidosis . It is recognized that due to the delivery mode of nystatin (i.e . topical, intermittent), as well as the cleansing effect of saliva within the oral environment, the yeasts undergo a relatively brief exposure to this drug during treatment . Nevertheless, there is a sparsity of data on the effect of such brief exposure to nystatin on the pathogenic attributes of Candida such as their adherence to host surfaces . The adhesion of microbes to host mucosal surfaces is a major determinant of successful colonization and infection . Thus the main aim of our investigation was to compare the in vitro adhesion of 30 oral isolates of Candida belonging to six different species (comprising Candida albicans, Candida tropicalis, Candida glabrata, Candida guilliermondii, Candida krusei and Candida parapsilosis) to human buccal epithelial cells, following their brief exposure (1 h) to minimum inhibitory concentration of nystatin, and subsequent removal of the drug . The adhesion of these isolates to buccal epithelial cells was assessed by a previously described adhesion assay . Compared with the controls, there was a significant reduction in buccal epithelial cell adhesion of all six Candida species after drug exposure (54%-68%) . However the adhesion of C . albicans isolates was the least affected by nystatin exposure, which was significantly different from that of the non-albicans species . These findings imply that sub-therapeutic levels of nystatin, which are likely to persist in the oral cavity during dosing intervals, may also be beneficial, as they inhibit candidal colonization . The significant difference in nystatin-induced suppression of adhesion between C . albicans and the non-albicans species investigated is a further testimonial for the pre-eminent virulence of the former species. Chemotherapy, 2000 Jul-Aug, 46(4), 267 - 74 Exposure to subtherapeutic concentrations of polyene antifungals suppresses the adherence of Candida species to denture acrylic; Egusa H et al.; BACKGROUND: The adherence of Candida species to denture acrylic is the initial event leading to Candida-associated denture stomatitis, with Candida albicans being the main aetiological agent . However, the increased incidence of immunocompromised patients in the community has resulted in the emergence of a number of non-albicans Candida species as causative agents of this disease, which is commonly managed by topically delivered polyene antifungals . Hence, we investigated the effect of the exposure of denture acrylic surfaces to nystatin and amphotericin B on the subsequent adhesion of six different Candida species . METHODS: Acrylic strips were exposed to subtherapeutic concentrations of the two polyenes for 30 min, and the adhesion of 4 isolates each of C . albicans, Candida glabrata, Candida guilliermondii, Candida krusei, Candida parapsilosis and Candida tropicalis was assessed using a previously described in vitro method with slight modifications . RESULTS: Overall, the results indicated a 35.9% (p < 0.01) and 63.1% (p < 0 . 01) reduction, respectively, in yeast adhesion to denture acrylic following exposure to nystatin and amphotericin B, although this effect was not uniform for all the tested isolates . Thus, all C . glabrata, 3 C . guilliermondii and a single isolate each of C . krusei, C . parapsilosis and C . tropicalis were not significantly affected by nystatin exposure, and a single isolate each of C . glabrata and C . guilliermondii were not significantly affected by amphotericin B . CONCLUSIONS: The present data, the first on the effect of polyenes on a wide range of Candida species, indicate that the in vitro exposure of denture acrylic to subtherapeutic concentrations of nystatin and amphotericin B suppresses the adherence of pathogenic Candida species in general . Chest, 2000 Jun, 117(6), 1672 - 8 Fungal empyema thoracis: an emerging clinical entity; Ko SC et al.; STUDY OBJECTIVES: To analyze the clinical spectra, pathogenesis, treatment, outcome, and prognostic factors of fungal empyema thoracis . DESIGN: The medical records of patients with positive fungal cultures from pleural effusions were retrospectively analyzed . SETTING: A university-based tertiary care hospital in Taipei, Taiwan . PATIENTS AND METHODS: From January 1990 through December 1997, patients diagnosed with fungal empyema were included in this study . The criteria for diagnosis of fungal empyema thoracis were as follows: (1) isolation of a fungal species from the pleural effusion; (2) significant signs of infection, such as fever (body temperature > 38.3 degrees C) and leukocytosis (white blood cell > 10,000/microL); and (3) isolation of the same mold species from pleural effusion on more than one occasion, or from pleural effusion and other specimens such as blood, sputum, or surgical wounds that showed evidence of tissue invasion . RESULTS: Sixty-seven patients with fungal empyema thoracis were included . Their mean age was 54 years (range, 2 weeks to 93 years), and 64% (43 patients) were men . Fifty-seven patients (85%) had various underlying diseases, and 18 (27%) had more than one immunocompromising condition . A total of 73 fungal isolates were recovered from pleural effusion; the most commonly encountered were Candida species (47 isolates, 64%), Torulopsis glabrata (13 isolates, 18%), and Aspergillus species (9 isolates, 12%) . Candida albicans (28 isolates) was the most common Candida species, followed by Candida tropicalis (13 isolates) . Six patients (9%) had two fungal strains isolated, and 16 (24%) had concomitant bacterial empyema thoracis . Eighteen patients (27%) had concurrent fungemia . Most (56 patients, 84%) cases of fungal empyema thoracis were nosocomial, and many case (43 patients, 64%) were acquired in ICUs . Abdominal disease (20 patients, 30%), especially previous abdominal surgery and GI perforation (12% and 10%, respectively), was the most common cause of fungal empyema thoracis, followed by bronchopulmonary infection (15 patients, 22%) and chest surgery (12 patients, 18%) . Forty-nine patients (73%) received systemic antifungal therapy, and 38 (57%) underwent closed drainage therapy . Eleven patients (16%) underwent pleural irrigation with normal saline solution, povidone-iodine solution, or antifungal agents . Six patients (9%) finally received decortication . All patients receiving surgery or pleural irrigation with antifungal agents survived . Despite the aforementioned management, the crude mortality was high (73%) . Multivariate analysis showed a significantly increased risk of death in immunocompromised patients (relative risk, 1.58; p < 0.005) and those with respiratory failure (relative risk, 2.31; p < 0.001) . Systemic antifungal therapy was associated with a significantly lower risk of death (relative risk, 0.69; p < 0.05) . CONCLUSION: These data imply an increasing incidence of fungal empyema thoracis in recent years and the necessity for aggressive treatment of patients with this disease. Antimicrob Agents Chemother, 2000 Jul, 44(7), 1917 - 20 Evaluation of voriconazole pharmacodynamics using time-kill methodology; Klepser ME et al.; Voriconazole is an investigational azole antifungal agent with activity against a variety of fungal species, including fluconazole-susceptible and -resistant Candida species and Cryptococcus neoformans . In this study, we employed in vitro time-kill methods to characterize the relationship between concentrations of voriconazole and its fungistatic activity against Candida albicans, Candida glabrata, Candida tropicalis, and C . neoformans . Isolates were exposed to voriconazole concentrations ranging from 0.0625 to 16 times the MIC, and the viable colony counts were determined over time . The 50 and 90% effective concentrations (EC(50) and EC(90), respectively) were determined at 8, 12, and 24 h following the addition of voriconazole . At each time point, near-maximal fungistatic activity, as indicated by the EC(90), was noted at a drug concentration of approximately three times the MIC . Additionally, EC(50) and EC(90) did not change over time, thus suggesting that the rate of activity was not improved by increasing concentrations . Voriconazole exhibits non-concentration-dependent pharmacodynamic characteristics in vitro. Int J Antimicrob Agents, 2000 Jul, 15(2), 83 - 90 Criteria used when initiating antifungal therapy against Candida spp . in the intensive care unit; Munoz P et al.; Invasive candidiasis is a life threatening complication for intensive care unit (ICU) patients . The infection is difficult to recognise so that treatment may be delayed or even not given . Risk factors for candidiasis include the use of antimicrobial agents, central intravascular devices (mainly Hickmann catheters), recurrent gastrointestinal perforations, surgery for acute pancreatitis or splenectomy and renal dysfunction or haemodialysis . Therapy against Candida spp is recommended in ICU patients with endophthalmitis or chorioretinitis possibly caused by Candida spp., in symptomatic patients with risk factors for invasive candidiasis especially if two or more anatomical sites are colonised and for asymptomatic high-risk surgical patients (with recent abdominal surgery or recurrent gastrointestinal perforations or anastomotic leakages) . The isolation of Candida from any site poses an increased risk but there are a few microbiological data that might help to establish the predictive value of a particular isolate . These include the site of isolation, the number of culture positive, noncontigous sites, the density of colonisation and the species isolated . Antifungals should be started when Candida spp . are recovered from blood cultures or from usually sterile body fluids, abscesses or wounds in burns patients . They should also be considered in patients with a colonisation index >0.5 or a corrected colonization index >0.4 or when the isolate is identified as Candida tropicalis. Int J Syst Evol Microbiol, 2000 May, 50 Pt 3, 1381 - 9 DNA fingerprinting patterns of Candida species using HinfI endonuclease; Fujita S et al.; Strain delineation was performed by means of restriction endonuclease analysis (REA) of genomic DNA with the restriction enzyme HinfI followed by conventional electrophoresis . A total of 337 yeast isolates representing 21 Candida species and five non-Candida yeast species was evaluated . A survey of isolates showed that Candida albicans and non-albicans species could be divided into mutually exclusive groups, and that subgroups could be created . Individual REA patterns for 111 C . albicans isolates, four Candida krusei isolates and 35 Candida glabrata isolates varied greatly, whereas 11 Candida dubliniensis isolates, 48 Candida tropicalis isolates and 41 Candida guilliermondii isolates could be divided into two, nine and nine groups, respectively . REA of the 49 Candida parapsilosis isolates with HinfI, however, showed that 47 (95 %) of them belonged to one group . REA patterns of the other yeast isolates, representing 19 species, were also quite different at the species level . These results showed that REA with HinfI may be useful for the identification and strain delineation of common and emerging Candida species. J Clin Microbiol, 2000 Jun, 38(6), 2254 - 60 Comparative evaluation of PASCO and national committee for clinical laboratory standards M27-A broth microdilution methods for antifungal drug susceptibility testing of yeasts; Arthington-Skaggs BA et al.; The PASCO antifungal susceptibility test system, developed in collaboration with a commercial company, is a broth microdilution assay which is faster and easier to use than the reference broth microdilution test performed according to the National Committee for Clinical Laboratory Standards (NCCLS) document M27-A guidelines . Advantages of the PASCO system include the system's inclusion of quality-controlled, premade antifungal panels containing 10, twofold serial dilutions of drugs and a one-step inoculation system whereby all wells are simultaneously inoculated in a single step . For the prototype panel, we chose eight antifungal agents for in vitro testing (amphotericin B, flucytosine, fluconazole, ketoconazole, itraconazole, clotrimazole, miconazole, and terconazole) and compared the results with those of the NCCLS method for testing 74 yeast isolates (14 Candida albicans, 10 Candida glabrata, 10 Candida tropicalis, 10 Candida krusei, 10 Candida dubliniensis, 10 Candida parapsilosis, and 10 Cryptococcus neoformans isolates) . The overall agreements between the methods were 91% for fluconazole, 89% for amphotericin B and ketoconazole, 85% for itraconazole, 80% for flucytosine, 77% for terconazole, 66% for miconazole, and 53% for clotrimazole . In contrast to the M27-A reference method, the PASCO method classified as resistant seven itraconazole-susceptible isolates (9%), two fluconazole-susceptible isolates (3%), and three flucytosine-susceptible isolates (4%), representing 12 major errors . In addition, it classified two fluconazole-resistant isolates (3%) and one flucytosine-resistant isolate (1%) as susceptible, representing three very major errors . Overall, the agreement between the methods was greater than or equal to 80% for four of the seven species tested (C . dubliniensis, C . glabrata, C . krusei, and C . neoformans) . The lowest agreement between methods was observed for miconazole and clotrimazole and for C . krusei isolates tested against terconazole . When the data for miconazole and clotrimazole were removed from the analysis, agreement was >/=80% for all seven species tested . Therefore, the PASCO method is a suitable alternative procedure for the testing of the antifungal susceptibilities of the medically important Candida spp . and C . neoformans against a range of antifungal agents with the exceptions only of miconazole and clotrimazole and of terconazole against C . krusei isolates. J Clin Microbiol, 2000 Jun, 38(6), 2219 - 26 Oral colonization, phenotypic, and genotypic profiles of Candida species in irradiated, dentate, xerostomic nasopharyngeal carcinoma survivors; Leung WK et al.; The aim of this study was to investigate oral yeast colonization and oral yeast strain diversity in irradiated (head and neck), dentate, xerostomic individuals . Subjects were recruited from a nasopharyngeal carcinoma clinic and were segregated into group A (age, <60 years {n = 25; average age +/- standard deviation (SD), 48 +/- 6 years; average postirradiation time +/- SD, 5 +/- 5 years}) and group B (age, >/=60 years {n = 8; average age +/- SD, 67 +/- 4 years; average postirradiation time +/- SD, 2 +/- 2 years}) and were compared with age- and sex-matched healthy individuals in group C (age, <60 years {n = 20; average age +/- SD, 44 +/- 12 years} and group D (age, >/=60 years {n = 10; average age, 70 +/- 3 years}) . Selective culture of oral rinse samples was carried out to isolate, quantify, and speciate yeast recovery . All test subjects underwent a 3-month comprehensive oral and preventive care regimen plus topical antifungal therapy, if indicated . A total of 12 subjects from group A and 5 subjects from group B were recalled for reassessment of yeast colonization . Sequential (pre- and posttherapy) Candida isolate pairs from patients were phenotypically (all isolate pairs; biotyping and resistotyping profiles) and genotypically (Candida albicans isolate pairs only; electrophoretic karyotyping by pulsed-field gel electrophoresis, restriction fragment length polymorphism {RFLP}, and randomly amplified polymorphic DNA {RAPD} assays) evaluated . All isolates were Candida species . Irradiated individuals were found to have a significantly increased yeast carriage compared with the controls . The isolation rate of Candida posttherapy remained unchanged . A total of 9 of the 12 subjects in group A and 3 of the 5 subjects in group B harbored the same C . albicans or Candida tropicalis phenotype at recall . Varying degrees of congruence in the molecular profiles were observed when these sequential isolate pairs of C . albicans were analyzed by RFLP and RAPD assays . Variations in the genotype were complementary to those in the phenotypic characteristics for some isolates . In conclusion, irradiation-induced xerostomia seems to favor intraoral colonization of Candida species, particularly C . albicans, which appeared to undergo temporal modifications in clonal profiles both phenotypically and genotypically following hygienic and preventive oral care which included topical antifungal therapy, if indicated . We postulate that the observed ability of Candida species to undergo genetic and phenotypic adaptation could strategically enhance its survival in the human oral cavity, particularly when salivary defenses are impaired. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1578 - 84 Experimental induction of fluconazole resistance in Candida tropicalis ATCC 750; Barchiesi F et al.; Candida tropicalis is less commonly isolated from clinical specimens than Candida albicans . Unlike C . albicans, which can be occasionally found as a commensal, C . tropicalis is almost always associated with the development of fungal infections . In addition, C . tropicalis has been reported to be resistant to fluconazole (FLC) . To analyze the development of FLC resistance in C . tropicalis, an FLC-susceptible strain (ATCC 750) (MIC = 1.0 microg/ml) was cultured in liquid medium containing increasing FLC concentrations from 8.0 to 128 microg/ml . The strain developed variable degrees of FLC resistance which paralleled the concentrations of FLC used in the medium . The highest MICs of FLC were 16, 256, and 512 microg/ml for strains grown in medium with 8.0, 32, and 128 microg of FLC per ml, respectively . Development of resistance was rapid and could be observed already after a single subculture in azole-containing medium . The resistant strains were cross-resistant to itraconazole (MIC > 1.0 microg/ml) and terbinafine (MIC > 512 microg/ml) but not to amphotericin B . Isolates grown in FLC at concentrations of 8.0 and 32 microg/ml reverted to low MICs (1.0 microg/ml) after 12 and 11 passages in FLC-free medium, respectively . The MIC for one isolate grown in FLC (128 microg/ml) (128 R) reverted to 16 microg/ml but remained stable over 60 passages in FLC-free medium . Azole-resistant isolates revealed upregulation of two different multidrug efflux transporter genes: the major facilitators gene MDR1 and the ATP-binding cassette transporter CDR1 . The development of FLC resistance in vitro correlated well with the results obtained in an experimental model of disseminated candidiasis . While FLC given at 10 mg/kg of body weight/day was effective in reducing the fungal burden of mice infected with the parent strain, the same dosing regimen was ineffective in mice infected with strain 128 R . Finally, the acquisition of in vitro FLC resistance in strain 128 R was related to a loss of virulence . The results of our study elucidate important characteristics and potential mechanisms of FLC resistance in C . tropicalis. Microbios, 2000, 102(401), 45 - 52 Growth of the fungal pathogen Candida in parotid saliva of patients with burning mouth syndrome; Chen Q et al.; Subclinical Candida infection has been suggested as one of the aetiological factors in patients with burning mouth syndrome (BMS) . In order to investigate the possible factors which contribute to the relatively high isolation rate of Candida in BMS, parotid saliva samples (20 in toto) from patients with this condition were collected and the growth of Candida in each sample dynamically observed using a computerized turbidometric assay system . A total of thirteen parotid saliva samples obtained from healthy individuals served as normal controls . The results showed no significant growth differential within the test and control saliva samples, when a single isolate each of Candida albicans and Candida tropicalis were cultured for 24 h, at 37 degrees C . A single isolate of Candida glabrata tended to grow better in the saliva from BMS patients than the controls . These results indicate that the composition of saliva may be a contributory factor for the high isolation rate of Candida in saliva of BMS patients. Zhonghua Wai Ke Za Zhi, 1997 Jul, 35(7), 395 - 7 {The nosocomial infection in ICU: characteristics and prevention}; An Y et al.; In 84 consecutive patients who were treated in ICU, 35 had microbiological 5 data . 117 strains (bacteria: 77; fungus: 40) were isolated from 24 of the 35 patients for 97 times . The positive rate was 68.6%, or 28.6% of 84 patients . The gram-negative bacilli were still the main source of nosocomial infection in ICU, especially the pseudomonas aeruginosa, clostridium welchii, and E . coli, but gram-positive cocci, especially MRSA infection were also increased . Mycotic infection increased very rapidly . 12 of the 24 patients were found a complicated mycotic infection . Six of the 12 (50%) died . Candida tropicalis and candida guilliermondi replaced candida albicans as the main pathogenic fungus . Sputum is still the most important specimen source of positive culture, followed by wound drainage, blood, urine, central venous catheter as well as feces . It is indicated that the respiratory system is still the main focus of nosocomial infection, and mycotic infection is most dangerous in ICU . Regular monitoring of microbial flora changes, detection correct use of antibiotics by drug sensitivity test, adequate surgical drainage, early and prevention of mycotic infection as well as improvement of ICU design will prevent or minimize the nosocomial infection in ICU. Kansenshogaku Zasshi, 2000 Mar, 74(3), 221 - 30 {Evaluation of a new method for antifungal drugs susceptibility testing to yeasts}; Ishigaki S et al.; We compared the Etest with a broth microdilution method (FP panel), performed according to the National Committee for modified Clinical Laboratory Standards (NCCLS) document M27-P guidelines, for determining the MICs of 81 clinical isolates of yeasts (7 Candida albicans, 8 Candida glabrata, 10 Candida parapsilosis, 6 Pichia anomala, 10 Candida tropicalis, 4 Candida guilliermondii, 4 Candida krusei, 6 Trichosporon cutaneum, 5 Candida ciferrii, 3 Candida famata, 4 Candida norvegensis, 2 Rhodotorula rubra, 3 Candida lusitaniae, 2 Candida curvata, 1 Candida inconspicua, 1 Candida intermedia, 1 Candida colliculosa, 1 Cryptococcus spp, 1 Tricosporon capitatum, 1 Pichia ohmeri, 1 Saccharomyces cerevisiae) . The Etest results for 6 ATCC standard strains correlated well with reference MICs except those of flucytosine (5-FC) for C . krusei, which tended to be 1 to 2 log2 dilution higher than the MIC range determined by NCCLS guidelines . However, the best agreement between the results for clinical isolates was seen with 5-FC (100% agreement {Within +/- 2 log2 dilutions} between the results of the two methods) . There was a 91.4% agreement between the results of the two methods with amphotericin B (Etest MICs tended to be 1 to 2 log2 dilution lower than those of the FP panel) . The Etest results with litraconazole for clinical isolates except C . tropicalis were similar to MICs of the FP panel (Etest for C . tropicalis showed 1 to 2 log2 dilution lower than FP panel) . Also, the Etest results with fluconazole for clinical isolates except C . tropicalis were similar of 1 log2 dilution higher than MICs of the FP panel (Etest for C . tropicalis showed more than 2 log2 dilution lower than FP panel) . These results showed a good level of overall agreement between the Etest method and the broth microdilution test (FP panel) . Since the Etest is a less laborintensive and much simpler method, it appears to be a useful procedure for testing the susceptibility of yeasts to antifungal agents. Can J Microbiol, 2000 Apr, 46(4), 350 - 7 Development of a xylitol biosensor composed of xylitol dehydrogenase and diaphorase; Takamizawa K et al.; In preparation for the development of a xylitol biosensor, the xylitol dehydrogenase of Candida tropicalis IFO 0618 was partially purified and characterized . The optimal pH and temperature of the xylitol dehydrogenase were pH 8.0 and 50 degrees C, respectively . Of the various alcohols tested, xylitol was the most rapidly oxidized, with sorbitol and ribitol being reduced at 65% and 58% of the xylitol rate . The enzyme was completely inactive on arabitol, xylose, glucose, glycerol, and ethanol . The enzyme's xylitol oxidation favored the use of NAD+ (7.9 U/mg) over NADP+ (0.2 U/mg) as electron acceptor, while the reverse reaction, D-xylulose reduction, favored NADPH (7.7 U/mg) over NADH (0.2 U/mg) as electron donor . The K(m) values for xylitol and NAD+ were 49.8 mM and 38.2 microM, respectively . For the generation of the xylitol biosensor, the above xylitol dehydrogenase and a diaphorase were immobilized on bromocyan-activated sephallose . The gel was then attached on a dissolved oxygen electrode . In the presence of vitamin K3, NAD+ and phosphate buffer, the biosensor recorded a linear response to xylitol concentration up to 3 mM . The reaction was stable after 15 min . When the biosensor was applied to a flow injection system, optimal operation pH and temperature were 8.0 and 30 degrees C, respectively . The strengths and limitations of the xylitol biosensor are its high affinity for NAD+, slow reaction time, narrow linear range of detection, and moderate affinity for xylitol. J Assoc Physicians India, 1999 Sep, 47(9), 921 - 2 Postoperative endocarditis due to Candida tropicalis; Mathew R et al.; In conclusion, we emphasize the importance of isolating the fungal agent from repeated blood cultures, particularly in infective endocarditis following heart surgeries. Antimicrob Agents Chemother, 2000 May, 44(5), 1242 - 6 In vitro activity of A-192411.29, a novel antifungal lipopeptide; Nilius AM et al.; A-192411.29 is a novel antifungal agent derived from the structural template of the natural product echinocandin . The in vitro activity of A-192411.29 against common pathogenic yeasts was assessed by National Committee for Clinical Laboratory Standards method M27-A . It demonstrated broad-spectrum, fungicidal activity and was active against the most clinically relevant yeasts, such as Candida albicans, Candida tropicalis, and Candida glabrata, as well as less commonly encountered Candida species; in general, its potency on a weight basis was comparable to that of amphotericin B . It maintained potent in vitro activity against Candida strains with reduced susceptibilities to fluconazole and amphotericin B . The in vitro activity of A-192411.29 against Cryptococcus neoformans was comparable to its activity against Candida spp . However, A-192411.29 did not demonstrate complete growth inhibition of Aspergillus fumigatus by the broth microdilution method used . A-192411.29 possesses an antifungal profile comparable to or better than those of fluconazole and amphotericin B against pathogenic yeasts, including strains resistant to fluconazole or amphotericin B, suggesting that it may be a therapeutically useful new antifungal drug. J Chemother, 2000 Feb, 12(1), 22 - 9 A head-on comparison of the in vitro antifungal activity of conventional and lipid-based amphotericin B: a multicenter study; Jessup C et al.; A comparative study of conventional amphotericin B, Abelcet and AmBisome was performed using a microdilution format of the NCCLS M27-A methodology for susceptibility testing against 300 fungal isolates (152 yeasts, 148 filamentous fungi) in both RPMI-1640 and antibiotic medium #3 (AB3) . The clinical isolates included Candida albicans (n=54), Candida glabrata (n=25), Candida parapsilosis (n=23), Candida krusei (n=19), Candida lusitaniae (n=14), Cryptococcus neoformans (n=5), Candida tropicalis (n=12), Aspergillus flavus (n=34), Aspergillus fumigatus (n=46) and 68 other filamentous fungi encompassing 22 different genera . The minimal inhibitory concentrations (MIC) for all drugs were defined as the lowest concentrations in which there was no visible growth . MICs were determined after 48 h for yeasts and 72 h for filamentous fungi . The mean MICs +/- standard error (microg/ml) for yeasts and filamentous fungi, respectively, were: Abelcet, 0.51+/-0.21, 4.34+/-0.61; AmBisome, 1.28+/-0.24, 5.68+/-0.57; amphotericin B, 0.29+/-0.11, 1.12+/-0.19, respectively . Overall, against both yeasts and filamentous fungi Abelcet proved to have more potent antifungal activity than AmBisome . Using AB3 as opposed to RPMI-1640 generally produced lower MIC values but did not have any effect on the order of relative activity with all of the antifungal agents tested . In conclusion, our data shows that Abelcet is more active than AmBisome against pathogenic yeast and filamentous fungi when assayed in AB3 in vitro . Comparison of the activities of these antifungals in experimental animal models is necessary to determine whether these in vitro findings are correlated with in vivo efficacy. Chemotherapy, 2000 May-Jun, 46(3), 209 - 12 Current status and fluconazole treatment of pelvic fungal gynecological infections; Mikamo H et al.; The incidence of opportunistic fungal infections has recently been increasing in many clinical fields . Fluconazole is commonly used against systemic fungal infections . The present study was undertaken to investigate the current status and the efficacy of fluconazole in pelvic fungal gynecological infections . Thirty-eight patients aged 36-72 years old diagnosed with pelvic peritonitis with positive fungal culture in pelvic ascites were enrolled in this study and given fluconazole treatment . Forty-two pathogens were isolated from the 38 assessable patients . The predominant pathogen was Candida albicans with an incidence of 61.9% (26/42) . Others included non-albicans Candida species amounting to 38.1% (16/42): 19.0% (8/42) Candida glabrata, 7.1% (3/42) Candida tropicalis, 7.1% (3/42) Candida parapsilosis and 4.8% (2/42) Candida krusei . The clinical cure rate at the end of fluconazole treatment was assessed as 30/38 (78.9%), and the fungal eradication rate as 26/42 (61.9%) . Each rate was 29/38 (76.3%) and 26/42 (61.9%), respectively, at 1 week after the treatment, while the eradication rate of C . albicans and non-albicans species was 20/26 (76.9%) and 6/16 (37.5%), respectively . There was no adverse effect except for slight elevations of GOT, GPT and LDH observed in 1 patient (2.6%), which returned to normal after the treatment . It seems there may be an increasing trend of non-albicans species in pelvic fungal gynecological infection, against which fluconazole appears to be rather effective . Arch Microbiol, 2000 Mar, 173(3), 187 - 92 A mutated hygromycin resistance gene is functional in the n-alkane-assimilating yeast Candida tropicalis; Hara A et al.; Development of a transformation system in the n-alkane-assimilating diploid yeast Candida tropicalis requires an antibiotic resistance gene in order to establish a selectable marker . The resistance gene for hygromycin B has often been used as a selectable marker in yeast transformation . However, C . tropicalis harboring the hygromycin resistance gene (HYG) was as sensitive to hygromycin B as the wild-type strain . Nine CTG codons were found in the ORF of the HYG gene . This codon has been reported to be translated as serine rather than leucine in Candida species . Analysis of the tRNA gene in C . tropicalis with the anticodon CAG {tRNA(CAG) gene}, which is complementary to the codon CTG, showed that the sequence was highly similar to that of the C . maltosa tRNA(CAG) gene . In C . maltosa, the codon CTG is read as serine and not leucine . These results suggested that the HYG gene was not functional due to the nonuniversal usage of the CTG codon . Each of the nine CTG codons in the ORF of the HYG gene was changed to a CTC codon, which is read as leucine, by site-directed mutagenesis . When a plasmid containing the mutated HYG gene (HYG#) was constructed and introduced into C . tropicalis, hygromycin-resistant transformants were successfully obtained . This mutated hygromycin resistance gene may be useful for direct selection of C . tropicalis transformants. J Bacteriol, 2000 May, 182(9), 2492 - 7 An n-alkane-responsive promoter element found in the gene encoding the peroxisomal protein of Candida tropicalis does not contain a C(6) zinc cluster DNA-binding motif; Kanai T et al.; When an asporogenic diploid yeast, Candida tropicalis, is cultivated on n-alkane, the expression of the genes encoding enzymes of the peroxisomal beta-oxidation pathway is highly induced . An upstream activation sequence (UAS) which can induce transcription in response to n-alkane (UAS(ALK)) was identified on the promoter region of the peroxisomal 3-ketoacyl coenzyme A (CoA) thiolase gene of C . tropicalis (CT-T3A) . The 29-bp region (from -289 to -261) present upstream of the TATA sequence was sufficient to induce n-alkane-dependent expression of a reporter gene . Besides n-alkane, UAS(ALK)-dependent gene expression also occurred in the cells grown on oleic acid . Several kinds of mutant UAS(ALK) were constructed and tested for their UAS activity . It was clarified that the important nucleotides for UAS(ALK) activity were located within 10-bp region from -273 to -264 (5'-TCCTGCACAC-3') . This region did not contain a CGG triplet and therefore differed from the sequence of the oleate-response element (ORE), which is a UAS found on the promoter region of 3-ketoacyl-CoA thiolase gene of Saccharomyces cerevisiae . Similar sequences to UAS(ALK) were also found on several peroxisomal enzyme-encoding genes of C . tropicalis. Am J Reprod Immunol, 2000 Mar, 43(3), 144 - 51 Polyherbal formulations with wide spectrum antimicrobial activity against reproductive tract infections and sexually transmitted pathogens; Talwar GP et al.; PROBLEM: Recent reports indicate high incidence of genital infections, most of which are sexually transmitted . Although specific drugs and antibiotics are available for some, a safe spermicidal formulation with wide spectrum antimicrobial action would be a desirable addition to the presently available spermicides . METHODS: Formulations at different dilutions were tested in culture systems on standard strains and clinical isolates including some isolates resistant to drugs . The effect on (HSV)-2 and Chlamydia trachomatis was determined in vivo in progestin sensitized mice . The effect on HIV-1 was investigated in two standardized systems . RESULTS: Polyherbal cream inhibited the growth in culture of clinical isolates of Candida albicans, Candida krusei and Candida tropicalis . Both the polyherbal cream and the Praneem polyherbal pessary inhibited urinary tract Escherichia coli (including multidrug resistant strains), and Neisseria gonorrhoeae (including 2 strains resistant to penicillin) . Both formulations manifested virucidal activity against HIV-1 at >2 and 50% dilutions (in two different test systems) on contact for 1-2 min . Intravaginal inoculation of the cream and the pessary suspensions before inoculation of the pathogen prevented lesions and vaginal transmission of HSV-2 and C . trachomatis in progestin sensitized mice . CONCLUSIONS: Polyherbal formulations have wide spectrum antibacterial, antifungal and antiviral effect against the tested sexually transmitted pathogens. Scand J Infect Dis, 2000, 32(1), 86 - 9 Successful non-surgical treatment of Candida tropicalis endocarditis with liposomal amphotericin-B (AmBisome); Melamed R et al.; Fungal endocarditis in children is most commonly a complication of palliative or curative surgery for congenital heart disease, rheumatic valvulitis and prolonged indwelling central venous and umbilical catheters . We describe here the case of a 3-y-old patient with chronic diarrhoea and prolonged total parenteral alimentation who developed severe C . tropicalis endocarditis and was treated successfully using a liposomal preparation of amphotericin-B (AmBisome) without surgical intervention. Drug Metabol Drug Interact, 1999, 15(2-3), 127 - 40 The in vivo metabolism of 5-(4-nitrophenyl)-4-phenyl-2,4-dihydro-3H-1,2,4-triazole-3-thione in rats; Oruc EE et al.; It is known that substituted 1,2,4-triazole-3-thione derivatives have several biological activities, such as antimicrobial, diuretic and antidepressant activities . In our previous studies, the antifungal activity of 5-(4-aminophenyl)-4-phenyl-2,4-dihydro-3H-1,2,4-triazole-3-thione was found to be active against Candida tropicalis K1022 . The aim of this study was to investigate the in vivo metabolic pathway of 5-(4-nitrophenyl)-4-phenyl-2,4-dihydro-3H-1,2,4-triazole-3-thione which was selected as a model compound for this study . The substrate and its potential metabolites, i.e . the acetylation and nitro reductive products, were synthesized and then separated using HPLC on a reverse phase system . In the in vivo metabolism study, a 4 mg dose was administered i.p . to male Wistar rats . Blood samples were collected at 0, 2, 4, 8, 12, 24 and 56 hours after administration and were passed through a Sep-Pak cartridge . The acetylated metabolite {5-(4-acetylaminophenyl)-4-phenyl-2,4-dihydro-3H-1,2,4-triazole-3-thione }, the amine metabolite {5-(4-aminophenyl)-4-phenyl-2,4-dihydro-3H-1,2,4-triazole-3-thione} and an unknown metabolite were detected. Eur J Clin Microbiol Infect Dis, 2000 Jan, 19(1), 61 - 3 Candida tropicalis vertebral osteomyelitis complicating epidural catheterisation with disease paralleled by elevated D-arabinitol/L-arabinitol ratios; Eisen DP et al.; Deep-seated Candida infections are challenging to diagnose by noninvasive means, and new modalities are needed to improve the yield of such investigations . Reported here is a case of Candida tropicalis vertebral osteomyelitis complicating epidural catheterisation in a diabetic patient with complicated abdominal sepsis . The diagnosis was supported by detection of increased D-arabinitol/L-arabinitol ratios in urine samples, and failure of medical management was indicated by elevated D-arabinitol/L-arabinitol ratios, which later decreased to baseline with successful surgical debridement and prolonged antifungal therapy. Diagn Microbiol Infect Dis, 2000 Feb, 36(2), 101 - 5 In vitro pharmacodynamic characteristics of flucytosine determined by time-kill methods; Lewis RE et al.; Two Candida albicans isolates, three non-albicans Candida isolates (Candida glabrata, Candida krusei, and Candida tropicalis), and one Cryptococcus neoformans isolate were evaluated by time-kill methods to characterize the relationship of flucytosine concentrations to antifungal activity and the duration of the post-antifungal effect (PAE) . Against Candida and Cryptococcusisolates, flucytosine at concentrations > 1 x MIC exhibited fungistatic (</=99% reduction in CFU) activity over a 24-h time-period . The rate and extent of fungistatic activity of flucytosine against all isolates was generally not increased when 5-FC concentrations exceeded 4 x MIC . A notable PAE was detected for flucytosine against both Candida and Cryptococcus species that persisted 2 to 4 h . These in vitro data suggest that flucytosine is predominately a concentration-independent fungistatic agent at clinically achieved serum concentrations . This pharmacodynamic characteristic coupled with the persistent PAE and the relatively long half-life of flucytosine in humans (> 5 h), suggests lower daily dosing may possible without loss of antifungal efficacy. Appl Environ Microbiol, 2000 Mar, 66(3), 930 - 6 Succession of microbial communities during hot composting as detected by PCR-single-strand-conformation polymorphism-based genetic profiles of small-subunit rRNA genes; Peters S et al.; A cultivation-independent technique for genetic profiling of PCR-amplified small-subunit rRNA genes (SSU rDNA) was chosen to characterize the diversity and succession of microbial communities during composting of an organic agricultural substrate . PCR amplifications were performed with DNA directly extracted from compost samples and with primers targeting either (i) the V4-V5 region of eubacterial 16S rRNA genes, (ii) the V3 region in the 16S rRNA genes of actinomycetes, or (iii) the V8-V9 region of fungal 18S rRNA genes . Homologous PCR products were converted to single-stranded DNA molecules by exonuclease digestion and were subsequently electrophoretically separated by their single-strand-conformation polymorphism (SSCP) . Genetic profiles obtained by this technique showed a succession and increasing diversity of microbial populations with all primers . A total of 19 single products were isolated from the profiles by PCR reamplification and cloning . DNA sequencing of these molecular isolates showed similarities in the range of 92.3 to 100% to known gram-positive bacteria with a low or high G+C DNA content and to the SSU rDNA of gamma-Proteobacteria . The amplified 18S rRNA gene sequences were related to the respective gene regions of Candida krusei and Candida tropicalis . Specific molecular isolates could be attributed to different composting stages . The diversity of cultivated bacteria isolated from samples taken at the end of the composting process was low . A total of 290 isolates were related to only 6 different species . Two or three of these species were also detectable in the SSCP community profiles . Our study indicates that community SSCP profiles can be highly useful for the monitoring of bacterial diversity and community successions in a biotechnologically relevant process. Infection, 2000 Jan-Feb, 28(1), 26 - 9 Epidemiology of candidemia--a nationwide survey in Israel; Rennert G et al.; Bloodstream infections with Candida are often lethal and have been reported to be increasing in frequency . The current retrospective study describes the magnitude and epidemiological characteristics of candidemia in all western-type hospital facilities in Israel in 1994 . Comprehensiveness of the data from the reporting hospitals was checked by cross-study of the data from the infectious diseases records and from the hospitalization records . Vital status of all reported cases was evaluated 1 year after the diagnosis . Data on 298 newly diagnosed cases of candidemia were received from 14 of the 18 general hospitals in Israel . The proportion of candidemia in the Israeli hospitals ranged from 0.1 to 0.01% of all admissions, with a mean of 0.05% . The incidence of candidemia differed significantly between the wards from 4-5/10,000 in general surgery and internal medicine wards to about 60/10,000 and 80/10,000 in intensive care and preterm units, respectively . Of all detected cases 53.6% were Candida albicans . Another nine specific species of Candida (mainly Candida parapsilosis, Candida tropicalis and Candida glabrata) were detected, with major differences between the various hospitals . The species of Candida differed significantly by sex and age . Of the cases of candidemia 21.5% died within 30 days of the isolation of the pathogen . The one-year mortality rate was 31.9% . Species-specific 30-day mortality rate was highest for C . glabrata . Throughout the analysis, C . glabrata emerged as a unique cause of candidemia, producing higher mortality, appearing at a younger age and predominating among females. Br J Biomed Sci, 1999, 56(2), 99 - 104 Characterisation of yeasts implicated in vulvovaginal candidosis in Irish women; al-Rawi N et al.; High-vaginal swabs were taken from 98 women attending a genitourinary clinic in Dublin city who presented with symptoms of vaginitis . Twenty-eight (28.6%) proved culture-positive for yeasts . Candida albicans was isolated from 26 of the yeast-positive patients and Candida glabrata and Candida tropicalis were isolated from a further two patients . Two patients were colonized by two yeasts simultaneously (C . albicans and Candida lucitaniae, and C . albicans and Candida krusei) . Yeasts were characterised on the basis of their adherence ability, extracellular enzyme production and resistance to antifungal agents . All C . albicans isolates demonstrated high adherence abilities to buccal epithelial cells, but produced relatively low levels of phospholipase and acid proteinase . The non-C . albicans isolates demonstrated low levels of adherence, but no extracellular enzyme production was detected . Isolates were most sensitive to the polyenes amphotericin B and nystatin but a large proportion (50%) of C . albicans isolates were resistant to clotrimazole, which is an important agent in the treatment of vulvovaginal candidosis. Fungal Genet Biol, 1999 Dec, 28(3), 135 - 45 Integrin-like proteins in Candida spp . and other microorganisms; Hostetter MK; The vertebrate integrins provide a paradigm for cell surface proteins involved in adhesion and morphogenesis . However, homologs of integrins have been found in more primitive organisms . This review will discuss the evidence for surface proteins in Candida albicans and Candida tropicalis that contain motifs reminiscent of integrins and will analyze the contributions of one of these proteins, Int1p, to adhesion, morphogenesis, and virulence . Other microorganisms thought to express integrin-like proteins will also be addressed . J Biol Chem, 2000 Feb 11, 275(6), 4445 - 52 A consensus sequence for long-chain fatty-acid alcohol oxidases from Candida identifies a family of genes involved in lipid omega-oxidation in yeast with homologues in plants and bacteria; Vanhanen S et al.; The yeast Candida cloacae is capable of growing on alkanes and fatty acids as sole carbon sources . Transfer of cultures from a glucose medium to one containing oleic acid induced seven proteins of M(r) 102,000, 73,000, 61,000, 54,000, and 46,000 and two in the region of M(r) 45,000 and repressed a protein of M(r) 64,000 . The induction of the M(r) 73,000 protein reached a 7-fold maximum 24 h after induction . The protein was confirmed by its enzyme activity to be a long-chain fatty-acid alcohol oxidase (LC-FAO) and purified to homogeneity from microsomes by a rapid procedure involving hydrophobic chromatography . An internal peptide of 30 amino acids was sequenced . A 1100-base pair cDNA fragment containing the LC-FAO peptide coding sequence was used to isolate a single exon genomic clone containing the full-length coding sequence of an LC-FAO (fao1) . The fao1 gene product was expressed in Escherichia coli and was translated as a functional long-chain alcohol oxidase, which was present in the membrane fraction . In addition, full-length coding sequences for a Candida tropicalis LC-FAO (faoT) and a second C . cloacae LC-FAO (fao2) were isolated . The DNA sequences obtained had open reading frames of 2094 (fao1), 2091 (fao2), and 2112 (faoT) base pairs . The derived amino acid sequences of fao2 and faoT showed 89.4 and 76.2% similarities to fao1 . The fao1 gene is much more highly induced on alkane than is fao2 . Although this study describes the first known DNA sequences encoding LC-FAOs from any source, there are unassigned Arabidopsis sequences and an unassigned Mycobacterium sequence in the GenBank(TM) Data Bank that show strong homology to the described LC-FAO sequences . The conservation of sequence between yeast, plants, and bacteria suggests that an as yet undescribed family of long-chain fatty-acid oxidases exists in both eukaryotes and prokaryotes. J Clin Microbiol, 2000 Feb, 38(2), 870 - 1 Trends in antifungal susceptibility among Candida sp . Urinary isolates from 1994 and 1998; Baran J Jr et al.; Antifungal susceptibilities were determined from 80 urinary isolates of Candida species collected in 1994 and 1998 . Our findings demonstrate increasing geometric means of fluconazole MICs and fluconazole resistance in Candida albicans and Candida tropicalis (those for Candida glabrata were unchanged) within the 4-year span . Amphotericin B and voriconazole MICs remained constant. J Clin Microbiol, 2000 Feb, 38(2), 537 - 41 Preliminary evaluation of a semisolid agar antifungal susceptibility test for yeasts and molds; Provine H et al.; This report presents a semisolid agar antifungal susceptibility (SAAS) method for the rapid susceptibility screening of yeasts and molds . The reproducibility and accuracy of the SAAS method were assessed by comparing the MICs of amphotericin B and fluconazole obtained for 10 candidate quality control (QC) American Type Culture Collection yeast strains in >/=15 replicates with those found by six independent laboratories using the National Committee for Clinical Laboratory Standards (NCCLS) M27-P broth macrodilution method (M . A . Pfaller et al., J . Clin . Microbiol . 33:1104-1107, 1995) . Overall, 96% of MICs for both drugs fell within 1 log(2) dilution of the modal MIC for each strain . The MICs for amphotericin B showed 99% agreement with the NCCLS proposed QC ranges within 1 log(2) dilution . Likewise, the MICs for fluconazole at >/=75% growth reduction showed 99% agreement for seven strains . Three strains, Candida albicans ATCC 24333 and ATCC 76615 and Candida tropicalis ATCC 750, showed a less sharp fluconazole endpoint at >/=75% growth reduction, but at >50% growth reduction, the agreement was 98% within 1 log(2) dilution of the proposed range . The MIC agreement within the proposed range for the suggested QC strains Candida parapsilosis ATCC 22019 and Candida krusei ATCC 6258 was 100% for fluconazole and 100% within 1 log(2) dilution of the proposed range for amphotericin B . The SAAS method demonstrated the susceptibility or resistance of 25 clinical isolates of filamentous fungi such as Aspergillus fumigatus to amphotericin B, itraconazole, and fluconazole, usually within 48 h . Although the results are preliminary, this SAAS method is promising as a rapid and cost-effective screen and is worthy of concerted investigation. Infect Immun, 2000 Feb, 68(2), 570 - 6 Adherence of platelets to Candida species in vivo; Robert R et al.; The in vivo interactions of platelets with Candida species yeast cells were investigated in a murine model . Mice were injected intravenously via the lateral caudal vein, and blood drawn by periorbital puncture was collected in phosphate-buffered saline-formaldehyde to avoid in vitro platelet activation . The study of the clearance of blastoconidia of Candida albicans and Candida glabrata showed that these cells disappeared quickly from the bloodstream . Microscopic observation of blood samples, stained by Calcofluor white or May Grunwald Giemsa, demonstrated the rapid attachment of platelets to fungal elements of all the Candida spp . tested . The attachment of murine platelets to C . albicans cells, observed by scanning electron microscopy, revealed morphological changes . The platelets lost their discoid shape, generated pseudopodia, and flattened against the yeast cells . The reversibility of platelet binding to C . albicans by chelating agents suggests a cation-dependent link . In contrast, the fixation of C . glabrata and Candida tropicalis was not modified by chelating agents . The mechanisms involved in the in vivo adherence of platelets to Candida cells may therefore differ according to the species of Candida. Int J Food Microbiol, 1999 Dec 15, 53(2-3), 169 - 84 Degradation of cyanogenic glycosides by Lactobacillus plantarum strains from spontaneous cassava fermentation and other microorganisms; Lei V et al.; Strains of Lactobacillus plantarum, Leuconostoc mesenteroides, Candida tropicalis and Penicillium sclerotiorum were screened for 19 enzymatic activities using the commercial kit API zym (Bio Merieux) . This activity was compared to the ability of degrading the toxic cyanogenic glycosides amygdalin, linamarin, and linseed cyanogens (a mixture of linustatin and neolinustatin) . Good correlation between the beta-glucosidase activity found in the API zym screening and the ability to degrade the cyanogenic glycosides was found for the first three species mentioned . P . sclerotiorum strains exhibited very high activity in the API zym test (substrate: 6-Br-2-naphthyl-beta-D-glucopyranoside), but proved unable to degrade any of the cyanogenic substrates . Among the seven strains of L . plantarum tested, a great variation was seen in the beta-glucosidase activity as well as in the ability