Candida tropicalis

Arch Esp Urol, 1996 Jan-Feb, 49(1), 66 - 8
{Candidiasis of the upper urinary tract . Report of a case}; Sanchez Sanchis M et al.; OBJECTIVE: We report a case of candidiasis of the upper urinary tract that presented as acute renal failure associated with septic syndrome . The patient initially required hemodialysis . Right hydronephrosis and perirenal collection were observed on ultrasound examination . METHODS: A percutaneous nephrostomy was performed . Nephrostomy urine cytology and cultures were positive for Candida tropicalis . An anterograde pyelography showed a 'fungus ball' in the urinary tract . RESULTS: Therapy with oral fluconazole and percutaneous amphotericin B achieved excellent results . CONCLUSIONS: Candidiasic urinary infection of the upper urinary tract often produces obstructive uropathy requiring percutaneous nephrostomy, which can also be used to instill amphotericin B . Combination therapy with amphotericin B and fluconazole can achieve excellent results.

Burns, 1995 Dec, 21(8), 594 - 6
Management of candida septicaemia in a regional burn unit; Still JM Jr et al.; Sepsis due to candida infection is a major cause of mortality and morbidity on our unit . Over a period of 3 years and 4 months, 29 cases of candida septicaemia, diagnosed by blood cultures, were encountered at the burn unit at Augusta Regional Medical Center . Factors known to predispose to fungal sepsis were present in all cases . All patients had large burns (14-98 per cent total body surface (TBSA) with a mean of 48.3 per cent) . All but one patient had at least one central venous line . Respiratory problems requiring ventilator support were present in 24 patients . Sixteen patients had Candida albicans sepsis, two in association with another fungal sepsis . Candida parapsilosis was encountered in nine patients, one in combination with another species . Four patients had Candida tropicalis . Amphotericin B was prescribed therapeutically in 25 patients, in seven together with fluconazole . Two patients received fluconazole only and two received no antifungal therapy . There were eight deaths all attributed to sepsis and all of whom had multiple organ failure . Five of those who died had completed a course of amphotericin B therapy, two were receiving treatment at the time of death, and one patient died before culture data became available . Early and aggressive therapy is advised and amphotericin B appears to be the drug of choice.

Arzneimittelforschung, 1995 Dec, 45(12), 1338 - 40
In vitro activity of clotrimazole for Candida strains isolated from recent patient samples; Schmidt A; Minimum inhibitory concentrations (MICs) of clotrimazole (CAS 23593-75-1, Bay 5097, clo) were determined for 142 clinical Candida isolates obtained between 1992 and 1994, including the species Candida albicans (96 strains), Candida glabrata (12 strains), Candida krusei (12 strains), and Candida tropicalis (12 strains) . For some of the Candida isolates of all four species, the MICs of amphotericin B and fluconazole were also determined . No MICs of clo of > 4 micrograms/ml were found for all four Candida species, the median of MICs of clo for Candida albicans being 0.03 micrograms/ml with a range from < 0.015 to 4 micrograms/ml . The MICs of clo for Candida albicans were on average 2 log stages below the MICs of fluconazole and 1 log step below the MICs of amphotericin B on the microgram/ml level . Taking account of the present in vitro resistance situation, clo is a highly active antimycotic substance for isolates of all Candida species with human pathogenetic relevance.

J Clin Microbiol, 1995 Dec, 33(12), 3216 - 20
PCR and single-strand conformational polymorphism for recognition of medically important opportunistic fungi; Walsh TJ et al.; The application of PCR technology to molecular diagnostics holds great promise for the early identification of medically important pathogens . PCR has been shown to be useful for the detection of the presence of fungal DNA in both laboratory and clinical samples . Considerable interest has been focused on the utility of selecting universal primers, those that recognize constant regions among most, if not all, medically important fungi . Once an amplicon, or piece of amplified DNA determined by the unique pair of oligonucleotide primers, has been generated, several different methods may be used to distinguish between genera and between species . The two major approaches have utilized differences in restriction enzyme digestion patterns or hybridization with specific probe . We report the application of single-strand conformational polymorphism (SSCP) as a technique to delineate the differences between fungal species and/or genera . Minor sequence variations in small single-stranded DNA cause subtle changes in conformation, allowing these strands to be separated on polyacrylamide gels by SSCP . We used a 197-bp fragment amplified from the 18S rRNA gene, common to all medically important fungi . After amplification, the fragments were denatured and run on an acrylamide-glycerol gel at room temperature or 4 degrees C for 4.5 or 4 h, respectively . Under room temperature conditions, the SSCP patterns for Candida albicans, Candida tropicalis, and Candida parapsilosis were identical and all strains within each species demonstrated the same pattern . These patterns differed markedly from those of the genus Aspergillus . The SSCP patterns of major and minor bands at room temperature permitted distinction between strains of Aspergillus fumigatus and Aspergillus flavus . There also was consistency of the SSCP banding pattern among different strains of the same Aspergillus species . The SSCP patterns for other medically important opportunistic fungi, such as Cryptococcus neoformans, Pseudallescheria boydii, and Rhizopus arrhizus, were sufficiently unique to permit distinction from those of C . albicans and A . fumigatus . We conclude that the technique of PCR-SSCP provides a novel method by which to recognize and distinguish medically important opportunistic fungi and which has potential applications to molecular diagnosis, taxonomic classification, molecular epidemiology, and elucidation of mechanisms of antifungal drug resistance.

Mycoses, 1995 Nov-Dec, 38(11-12), 443 - 8
Testing susceptibility of Candida species to fluconazole and itraconazole using the microdilution assay; Seibold M et al.; A microdilution system was established for testing the susceptibility of Candida species to fluconazole and itraconazole . The assay used a sodium phosphate-buffered (0.1 mol l-1) Casitone/glucose medium (pH 7.2) supplemented with potassium, iron, magnesium, trace elements and vitamins . Tests were read photometrically after 24 h, and the minimum inhibitory concentration (MIC) was defined as the IC90 . In nearly all strains sharp end points were observed . The MICs against Candida species without any known pre-exposure to azoles were found to range from 0.2 to 1.56 micrograms ml-1 for fluconazole and from 2.3 to 12 ng ml-1 for intraconazole . For strains from candidosis patients refractory to treatment with fluconazole the MICs of fluconazole ranged from 6.25 to 100 micrograms ml-1, while those for itraconazole varied between 12 and > 450 ng ml-1 . The strains did not respond congruent to both azoles . A similar disparity of the MICs was observed with Candida tropicalis and Candida krusei . The unusually low MICs of itraconazole were attained because (1) the dilution series was prepared from a preformed (concentrated) dilution series in 75% dimethylsulphoxide and not directly by serial dilution in the broth and (2) the incubation was performed in microtitre plates made of quartz glass rather than in the generally used polystyrene microtitre plates.

J Antimicrob Chemother, 1995 Nov, 36(5), 787 - 93
Itraconazole susceptibilities of fluconazole susceptible and resistant isolates of five Candida species; Johnson EM et al.; The in-vitro susceptibilities of 1380 isolates of five Candida species were determined in order to establish whether isolates resistant to fluconazole were cross-resistant to itraconazole . IC50 values were determined by a broth microdilution method . 690 Candida albicans isolates, seven Candida glabrata isolates, seven Candida krusei isolates, 120 Candida parapsilosis isolates and 37 Candida tropicalis isolates were susceptible to both fluconazole (IC50 < or = 32 mg/L) and itraconazole (IC50 < or = 4 mg/L) . Twenty eight of 160 C . albicans isolates (17.5%), 180 of 293 C . glabrata isolates (61.4%), six of 48 C . krusei isolates (12.5%), and 10 of 18 C . tropicalis isolates (55.5%) resistant to fluconazole (IC50 > or = 64 mg/L) were also resistant to itraconazole (IC50 > or = 8 mg/L) . In contrast, drug-specific resistance to itraconazole was not observed in any of the isolates tested . However, the itraconazole IC50s for fluconazole susceptible isolates were lower than those for fluconazole resistant isolates, which suggests that patients who fail fluconazole treatment might require itraconazole at higher dosages than usual.

J Clin Microbiol, 1995 Nov, 33(11), 3025 - 7
Use of CHROMagar Candida medium for isolation of yeasts from dental samples; Beighton D et al.; A new differential medium, CHROMagar Candida, for the isolation of clinically important yeasts was investigated to determine its usefulness in facilitating the study of oral yeasts . The recovery of yeasts on the medium was not significantly different from the recovery on Sabouraud dextrose agar . The identities of 450 green colonies on CHROMagar Candida, presumptively identified as Candida albicans on the basis of the manufacturer's instructions, were confirmed by testing for beta-N-acetylgalactosaminidase . Candida tropicalis also formed distinctive colonies, and other yeasts including Candida (Torulopsis) glabrata, Candida Parapsilosis, Candida Magnoliae, Candida lusitaniae, Candida Famata, Candida kefir, and Saccharomyces cerevisiae were readily distinguished from C . albicans and C . tropicalis isolates . CHROMagar Candida is a very useful medium, and its use will facilitate the study of yeasts associated with dental diseases.

J Clin Microbiol, 1995 Nov, 33(11), 2913 - 9
Molecular probes for diagnosis of fungal infections; Sandhu GS et al.; We have developed 21 specific nucleic acid probes which target the large subunit rRNA genes from Aspergillus flavus, Aspergillus fumigatus, Aspergillus glaucus, Aspergillus niger, Aspergillus terreus, Blastomyces dermatitidis, Candida albicans, Candida (Torulopsis) glabrata, Candida guilliermondii, Candida kefyr, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, Coccidioides immitis, Cryptococcus neoformans var . gattii, Cryptococcus neoformans var . neoformans, Filobasidiella neoformans var . bacillispora, Filobasidiella neoformans var . neoformans, Histoplasma capsulatum, Pseudallescheria boydii, and Sporothrix schenckii . A section of the 28S rRNA gene from approximately 100 fungi, representing about 50 species of pathogens and commonly encountered saprophytes, was sequenced to develop universal PCR primers and species-specific oligonucleotide probes . Each step in the process of detection and identification was standardized to a common set of conditions applicable without modification to all fungi of interest and all types of clinical specimens . These steps consist of DNA extraction by boiling specimens in an alkaline guanidine-phenol-Tris reagent, amplification of a variable region of the 28S rRNA gene with universal primers, and amplicon identification by probe hybridization or DNA sequencing performed under conditions identical for all fungi . The results obtained by testing a panel of fungal isolates and a variety of clinical specimens indicate a high level of specificity.

Biochem J, 1995 Oct 15, 311 ( Pt 2), 437 - 43
Molecular cloning of a novel widely expressed human 80 kDa 17 beta-hydroxysteroid dehydrogenase IV; Adamski J et al.; Reactions of oestrogens and androgens at position C-17 are catalysed by 17 beta-hydroxysteroid dehydrogenases (17 beta-HSDs) . Cloning of the cDNA of a novel human 17 beta-HSD IV and expression of its mRNA are described . A probe derived from the recently discovered porcine 17 beta-oestradiol dehydrogenase (17 beta-EDH) was used to isolate a 2.6 kb human cDNA encoding a continuous protein of 736 amino acids of high (84%) similarity to the porcine 17 beta-EDH . The calculated molecular mass of the human enzyme is 79,595 Da . Other sequence similarities shared by the two enzymes are: an N-terminal sequence which is similar to that of members of the short-chain alcohol dehydrogenase family; amino acids 343-607 which are similar to the C-terminal domains of a trifunctional Candida tropicalis enzyme and the FOX2 gene product of Saccharomyces cerevisiae; amino acids 596-736 which are similar to human sterol carrier protein 2 . The previously cloned human 17 beta-HSD I, II and III are less than 25% identical with 17 beta-HSD IV . mRNA for HSD IV is a single species of 3.0 kb, present in many tissues with highest concentrations in liver, heart, prostate and testes . When over-expressed in mammalian cells, the human 17 beta-HSD IV enzyme displays a specific unidirectional oxidative 17 beta-HSD activity.

Clin Microbiol Rev, 1995 Oct, 8(4), 462 - 78
New and emerging yeast pathogens; Hazen KC; The most common yeast species that act as agents of human disease are Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis, and Cryptococcus neoformans . The incidence of infections by other yeasts has increased during the past decade . The most evident emerging pathogens are Malassezia furfur, Trichosporon beigelii, Rhodotorula species, Hansenula anomala, Candida lusitaniae, and Candida krusei . Organisms once considered environmental contaminants or only industrially important, such as Candida utilis and Candida lipolytica, have now been implicated as agents of fungemia, onychomycosis, and systemic disease . The unusual yeasts primarily infect immunocompromised patients, newborns, and the elderly . The role of central venous catheter removal and antifungal therapy in patient management is controversial . The antibiograms of the unusual yeasts range from resistant to the most recent azoles and amphotericin B to highly susceptible to all antifungal agents . Current routine methods for yeast identification may be insufficient to identify the unusual yeasts within 2 days after isolation . The recognition of unusual yeasts as agents of sometimes life-threatening infection and their unpredictable antifungal susceptibilities increase the burden on the clinical mycology laboratory to pursue complete species identification and MIC determinations . Given the current and evolving medical practices for management of seriously ill patients, further evaluations of the clinically important data about these yeasts are needed.

J Antimicrob Chemother, 1995 Oct, 36(4), 713 - 6
In-vitro antifungal activity of sertaconazole, econazole, and bifonazole against Candida spp; Carrillo-Munoz AJ et al.; The in-vitro activity of sertaconazole (7-chloro-3-{1-(2,4-dichlorophenyl- 1-yl)ethoxy-methyl} benzo{b} thiophene) was compared with that of econazole and bifonazole against 150 strains of yeasts which includes six Candida species . Minimum inhibitory concentrations (MICs) were determined by a microdilution technique in Sabouraud's liquid medium, (pH 5.6) . Sertaconazole (arithmetic mean MIC 0.77 mg/L) was more active than econazole (MIC 1.75 mg/L) and bifonazole (MIC 9.05 mg/L) . MIC values for sertaconazole were generally and specifically lower for each tested species, Candida parapsilosis being the most susceptible (MIC 0.31 mg/L), in contrast to Candida tropicalis, which had the highest MIC (1.67 mg/L).

J Clin Microbiol, 1995 Oct, 33(10), 2660 - 4
Comparative evaluation of macrodilution and alamar colorimetric microdilution broth methods for antifungal susceptibility testing of yeast isolates; To WK et al.; A comparative evaluation of the macrodilution method and the Alamar colorimetric method for the susceptibility testing of amphotericin B, fluconazole, and flucytosine was conducted with 134 pathogenic yeasts . The clinical isolates included 28 Candida albicans, 17 Candida tropicalis, 15 Candida parapsilosis, 12 Candida krusei, 10 Candida lusitaniae, 9 Candida guilliermondii, 18 Torulopsis glabrata, and 25 Cryptococcus neoformans isolates . The macrodilution method was performed and interpreted according to the recommendations of the National Committee for Clinical Laboratory Standards (document M27-P), and the Alamar colorimetric method was performed according to the manufacturer's instructions . For the Alamar colorimetric method, MICs were determined at 24 and 48 h of incubation for Candida species and T . glabrata and at 48 and 72 h of incubation for C . neoformans . The overall agreement within +/- 1 dilution for Candida species and T . glabrata against the three antifungal agents was generally good, with the values for amphotericin B, fluconazole, and flucytosine being 85.3, 77.9, and 86.2%, respectively, at the 24-h readings and 69.3, 65.2, and 97.2%, respectively, at the 48-h readings . Most disagreement was noted with fluconazole against C . tropicalis and T . glabrata . Our studies indicate that determination of MICs at 24 h by the Alamar colorimetric method is a valid alternate method for testing amphotericin B, fluconazole, and flucytosine against Candida species but not for testing fluconazole against C . tropicalis and T . glabrata . For flucytosine, much better agreement can be demonstrated against Candida species and T . glabrata at the 48-h readings by the Alamar method . Excellent agreement within +/- dilution can also be observed for amphotericin B, fluconazole, and flucytosine (80, 96, and 96%, respectively) against c . neoformans when the MICs were determined at 72 h by the Alamar method.

Antimicrob Agents Chemother, 1995 Sep, 39(9), 2051 - 60
Antifungal susceptibility testing of yeasts: evaluation of technical variables for test automation; Odds FC et al.; The technical parameters for antifungal susceptibility testing with Candida species were reexamined to determine the optimal conditions for testing with semiautomated preparations of broth microdilution cultures, automated spectrophotometric readings of the cultures, and dose-response and endpoint determinations by means of a computer spreadsheet . Tests were based on proposed standard method M27P of the National Committee for Clinical Laboratory Standards for antifungal agents . RPMI 1640 broth with extra glucose to a final concentration of 2% gave higher and more reproducible drug-free control readings without affecting susceptibility endpoint readings . An inoculum of 8 x 10(4) yeasts per ml prepared from a carbon-limiting broth culture without further standardization was found to give optimal control readings after 48 h of incubation at 37 degrees C . For flucytosine, fluconazole, itraconazole, and ketoconazole, endpoints based on 50% growth inhibition (50% inhibitory concentration) gave the minimum variation with inoculum size and the fewest endpoint differences with RPMI 1640 medium obtained from two different suppliers . The 50% inhibitory concentration was also the optimal endpoint for fluconazole and ketoconazole susceptibilities in comparison with broth macrodilution MICs determined by the method of the National Committee for Clinical Laboratory Standards . Intralaboratory reproducibility was determined by retrospective analysis of replicate results for isolates retested at random over a 2-year period . This approach showed less favorable reproducibility than has been reported from purpose-designed, prospective antifungal susceptibility studies, but it may better reflect real-life test reproducibility . Susceptibility data for 616 clinical isolates of yeasts, representing 16 Candida and Saccharomyces spp., confirmed the tendency of Candida lusitaniae isolates to show relatively low susceptibilities to amphotericin B, the tendency of Candida krusei isolates to show low flucytosine and fluconazole susceptibilities, and the presence of some isolates in the species Candida albicans, Candida glabrata, and Candida tropicalis with low susceptibilities to azole derivative antifungal agents . The study demonstrates the value of automation and standardization in all stages of yeast susceptibility testing, from plate preparation to data analysis.

J Clin Microbiol, 1995 Sep, 33(9), 2476 - 9
Identification of Candida species by PCR and restriction fragment length polymorphism analysis of intergenic spacer regions of ribosomal DNA; Williams DW et al.; The PCR was used to amplify a targeted region of the ribosomal DNA from 84 Candida isolates . Unique product sizes were obtained for Candida guilliermondii, Candida (Torulopsis) glabrata, and Candida pseudotropicalis . Isolates of Candida albicans, Candida tropicalis, Candida stellatoidea, Candida parapsilosis, and Candida krusei could be identified following restriction digestion of the PCR products.

J Med Vet Mycol, 1995 Jul-Aug, 33(4), 275 - 8
pH-dependent denaturation of extracellular aspartic proteinases from Candida species; Wagner T et al.; The yeasts Candida albicans . Candida tropicalis, and Candida parapsilosis secrete aspartic proteinases which may enhance virulence . Profiles of pH-dependent irreversible denaturation of such enzymes were determined at 37 degrees C . C . albicans proteinases from both serotypes A and B maintained 50% of their activity near pH 7.25 . Proteinases from C . parapsilosis and C . tropicalis lost 50% of their activity at pH 6.75 and pH 6.15, respectively . This suggests that in the infected host only proteinases of C . albicans maintain a native state for any length of time.

J Med Vet Mycol, 1995 Jul-Aug, 33(4), 241 - 6
Random amplified polymorphic DNA for strain delineation within Candida tropicalis; Lin D et al.; Candida tropicalis DNA was used as a template in a polymerase chain reaction (PCR) utilizing a 10-mer primer to generate random amplified polymorphic DNA (RAPD) . RAPD patterns associated with 25 primers were obtained for six epidemiologically-unrelated isolates, then a subset of six primers were selected to screen a panel of 18 isolates of C . tropicalis and six isolates of Candida paratropicalis, a species that resembles C . tropicalis but has a sucrose-negative phenotype . The panel, which included nine epidemiologically-related isolates from an outbreak of sternal wound infections, was typed without knowledge of each isolate's origin . The RAPD profiles of the epidemiologically-related isolates were identical to very similar; in contrast, the profiles of most unrelated isolates showed more dissimilarity . While RAPD profiles of C . albicans and Candida parapsilosis differed substantially from those of C . tropicalis, the profiles obtained for C . paratropicalis were consistent with it being a variant of C . tropicalis.

Infect Immun, 1995 Jul, 63(7), 2714 - 9
Antibody response that protects against disseminated candidiasis; Han Y et al.; We previously showed that surface mannans of Candida albicans function as adhesins during yeast cell attachment to mouse splenic marginal zone macrophages . The mannan adhesin fraction was encapsulated into liposomes and used to vaccinate mice over a 5- to 6-week period . Circulating agglutinins specific for the fraction correlated with increased resistance to disseminated candidiasis . Antiserum from vaccinated animals protected naive BALB/cByJ mice against C . albicans serotype A and B strains and Candida tropicalis . Antiserum also protected SCID mice against disseminated disease . The serum protective ability was stable at 56 degrees C, but this ability was adsorbed by C . albicans cells . The antiserum was divided into three fractions after separation by high-performance liquid chromatography . One fraction contained all of the agglutinin activity and transferred resistance to naive mice . A second fraction also transferred resistance . Two monoclonal antibodies (MAbs) specific for candidal surface determinants were obtained . MAb B6.1 is specific for a mannan epitope in the adhesin fraction, and MAb B6 is specific for a different epitope in the fraction . Both MAbs are immunoglobulin M, and both strongly agglutinate candidal cells, but only MAb B6.1 protected both normal and SCID mice against disseminated candidiasis . In one experiment, 10 normal mice were given MAb B6.1 and challenged with yeast cells . Six mice survived the 67-day observation period; 4 of the survivors were cured as evidenced by the lack of CFU in the kidney and spleen . Our studies show that antibodies against certain cell surface antigens of C . albicans help the host resist disseminated candidiasis.

Appl Microbiol Biotechnol, 1995 Jul, 43(3), 489 - 92
A novel promoter, derived from the isocitrate lyase gene of Candida tropicalis, inducible with acetate in Saccharomyces cerevisiae; Umemura K et al.; When the isocitrate lyase gene, containing 5'-upstream and 3'-flanking regions, of an n-alkane-assimilating yeast Candida tropicalis was introduced into Saccharomyces cerevisiae, the enzyme was functionally overexpressed in the cells grown on acetate . The amount of the recombinant isocitrate lyase expressed in S . cerevisiae was as much as 30% of the total soluble proteins in the cells, being comparable to that with GAL7 functional under the control of galactose . The expression was also observed when the cells were grown on glycerol, lactate, ethanol or oleate . These facts indicate that the isocitrate lyase gene upstream region (UPR-ICL) contains a strong promoter functional in S . cerevisiae . UPR-ICL is active as a promoter on cheap carbon sources such as acetate and nonconventional carbon sources such as oleate, whereas many conventional strong promoters demand relatively expensive sugars or sugar derivatives . Therefore, it is promising to construct an economical recombinant protein production system by using UPR-ICL.

Antimicrob Agents Chemother, 1995 Jul, 39(7), 1517 - 21
In vitro and in vivo antifungal activities of DU-6859a, a fluoroquinolone, in combination with amphotericin B and fluconazole against pathogenic fungi; Nakajima R et al.; DU-6859a is an investigational fluoroquinolone agent with potent bactericidal activity, but by itself it has no antifungal activity . When combined with amphotericin B (AmB), however, DU-6859a clearly enhanced the in vitro antifungal activity of AmB against Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, and Cryptococcus neoformans in microdilution checkerboard studies . Positive interactions of DU-6859a with AmB against Aspergillus fumigatus were dependent on the medium used; yeast nitrogen base supplemented with amino acids, ammonium sulfate, and 1% glucose was better for demonstrating synergism, while in RPMI 1640 medium, unexpected antagonism between the drugs occurred against three of the strains tested . In combination with fluconazole (Flu), DU-6859a increased the activity of Flu against C . albicans both in synthetic amino acid medium fungal and in supplemented yeast nitrogen base . An in vitro time-kill study revealed that DU-6859a combined with AmB significantly suppressed the regrowth of C . albicans compared with the suppression brought about by AmB used alone in a concentration-dependent fashion . Furthermore, in a model of C . albicans infection in mice, the fungal load in infected kidneys was significantly less in mice given the combination treatment of DU-6859a plus either AmB or Flu, and thus, the combination treatment resulted in prolonged survival of infected mice compared with treatment with either antifungal alone . The prolonged survival in mice given the combined treatment was also observed in mice with A . fumigatus infection, indicating that DU-6859a potentiated the actions of the antifungal agents in vivo as well as in vitro.

J Infect Dis, 1995 Jun, 171(6), 1660 - 3
Epithelial adhesion in yeast species: correlation with surface expression of the integrin analog; Bendel CM et al.; Epithelial adhesion and expression of the integrin analog, a putative candidal adhesion, were correlated for 33 clinical and laboratory isolates of Candida albicans, other Candida species, and Saccharomyces cerevisiae . On flow cytometry with saturating concentrations of the monoclonal antibody OKM1, surface fluorescence was highest for C . albicans at 67.8% +/- 1.7% and significantly reduced for Candida tropicalis (32.0% +/- 2.6%), Candida parapsilosis (18.3% +/- 2.4%), Candida glabrata (3.3% +/- 0.8%), Candida lusitaniae (2.9% +/- 1.0%), Candida krusei (0.7% +/- 0.1%), and Saccharomyces cerevisiae (1.7% +/- 0.2%) (P < .006 for all other species vs . C . albicans) . Adhesion to a human epithelial cell line was highest for C . albicans at 49.8% +/- 3.5%, lower for C . tropicalis (44.7% +/- 4.6%), and incrementally reduced for all other species (< 25%) (P < .012) . The correlation between integrin expression and epithelial adhesion was highly significant (P = .0066; r2 = .8) . Surface expression of the integrin analog predicts epithelial adhesion for yeast species isolated in opportunistic infections.

Intern Med, 1995 Jun, 34(6), 485 - 90
Clinical evaluation of catheter-related fungemia and bacteremia; Inoue Y et al.; Forty-four patients with catheter-related infection admitted to Hokusho Central Hospital between 1985 and 1991 were studied retrospectively . The rate of catheter-related fungemia or bacteremia to all corresponding cases of fungemia and bacteremia increased from 7.7% in 1985 to 28.8% in 1991 . The isolated pathogens were Candida parapsilosis (8 strains), Candida tropicalis (6 strains), methicillin-resistant Staphylococcus aureus (MRSA) (6 strains), methicillin-sensitive S . aureus (MSSA) (5 strains) and Streptococcus epidermidis (3 strains) . Bacteremia occurred after catheterization of the femoral vein for a mean duration of 37 days . The period was significantly shorter than that after catheterization of the subclavian vein (56 days) . The major isolates from the subclavian vein were Candida spp . (14/17, 82.4%), followed by MRSA (1/17, 5.9%) and MSSA (1/17, 5.9%), while isolates from the femoral vein were Candida spp . (6/16, 37.5%), MRSA (5/16, 31.3%) and MSSA (3/16, 20.8%) . Catheter removal alone did not improve the clinical condition, particularly in MRSA bacteremia; the combination of antimicrobial therapy and removal of the catheter was necessary for a better prognosis.

Burns, 1995 May, 21(3), 167 - 70
A comparison of susceptibility to five antifungal agents of yeast cultures from burn patients; Still JM Jr et al.; Patients with significant degrees of immunocompromise, such as cancer, AIDS and large burns, who have received significant amounts of antibiotics, may develop infections with yeast organisms . Over a 3-year period, all patients with positive fungal blood cultures and most wounds of patients with large burns considered to be a risk of yeast infection were selected and tested for their susceptibility to five antifungal agents, amphotericin B, ketoconazole, miconazole, diflucan, and 5-fluorocytosine . In all, 244 specimens of yeast were tested: 142 Candida albicans, 52 Candida parapsilosis, 26 Candida tropicalis and 13 Trichosporon beigelii . A limited number of other isolates of Candida (12) were also encountered . All Candida organism were sensitive to amphotericin B . There was wide variation in regard to the susceptibility to the other four agents, with C . albicans and C . tropicalis being largely resistant to miconazole and ketoconazole . T . beigelii was recovered in 13 patients . One-half of these organisms was resistant to amphotericin B . Awareness of variations in species and susceptibility are helpful in the selection of appropriate therapeutic antifungal agents.

Arch Microbiol, 1995 May, 163(5), 322 - 8
The upstream region of the isocitrate lyase gene (UPR-ICL) of Candida tropicalis induces gene expression in both Saccharomyces cerevisiae and Escherichia coli by acetate via two distinct promoters; Atomi H et al.; The upstream region of the isocitrate lyase gene (UPR-ICL, 1530bp) of an n-alkane-utilizable yeast, Candida tropicalis, induced gene expression in another yeast, Saccharomyces cerevisiae, when the yeasts were grown on acetate . Surprisingly, UPR-ICL displayed the same regulatory function in the bacterium Escherichia coli when grown on acetate . We determined the interesting nucleotide sequence of UPR-ICL . The deletion analysis of UPR-ICL in both cells revealed the presence of two distinct promoters: one was localized at -394 to -379 and regulated gene expression in S . cerevisiae; the other was located near the initiation codon and regulated gene expression in E . coli . The two promoter sequences were similar, but not identical to regulatory elements that have been previously reported in S . cerevisiae and E . coli, respectively . Accordingly, the possibility of novel regulatory mechanisms could not be excluded . This is an interesting example of the presence of distinct cis-acting regulatory elements responsible for the induction of gene expression in one gene by acetate in both S . cerevisiae and E . coli . Preservation of such promoters through evolution is also discussed.

Mol Gen Genet, 1995 Apr 10, 247(1), 95 - 104
Molecular cloning, sequencing and sequence analysis of the fox-2 gene of Neurospora crassa encoding the multifunctional beta-oxidation protein; Fossa A et al.; We present the molecular cloning and sequencing of genomic and cDNA clones of the fox-2 gene of Neurospora crassa, encoding the multifunctional beta-oxidation protein (MFP) . The coding region of the fox-2 gene is interrupted by three introns, one of which appears to be inefficiently spliced out . The encoded protein comprises 894 amino acid residues and exhibits 45% and 47% sequence identity with the MFPs of Candida tropicalis and Saccharomyces cerevisiae, respectively . Sequence analysis identifies three regions of the fungal MFPs that are highly conserved . These regions are separated by two segments that resemble linkers between domains of other MFPs, suggesting a three-domain structure . The first and second conserved regions of each MFP are homologous to each other and to members of the short-chain alcohol dehydrogenase family . We discuss these homologies in view of recent findings that fungal MFPs contain enoyl-CoA hydratase 2 and D-3-hydroxyacyl-CoA dehydrogenase activities, converting trans-2-enoyl-CoA via D-3-hydroxyacyl-CoA to 3-ketoacyl-CoA . In contrast to its counterparts in yeasts, the Neurospora MFP does not have a C-terminal sequence resembling the SKL motif involved in protein targeting to microbodies.

Methods Find Exp Clin Pharmacol, 1995 Apr, 17(3), 163 - 7
Influence of free or liposomal amphotericin B on killing of Candida species by human peritoneal macrophages; Allwinn R et al.; The influence of free and liposomal amphotericin B at subinhibitory and inhibitory concentrations on killing of Candida albicans (ATCC 10231), Candida tropicalis (ATCC 13803) and Cryptococcus neoformans (930) by human peritoneal macrophages was investigated in vitro . Peritoneal macrophages were harvested from overnight peritoneal dialysate of 10 patients undergoing regular continuous ambulatory peritoneal dialysis and incubated with Candida species (1:2), pooled human serum, fetal calf serum, Medium 199 and Sabouraud broth (with or without amphotericin B) for 6 hours . The killing of Candida species was determined using subculture technique . The combination of amphotericin B (free or liposomal) with peritoneal macrophages enhanced the killing of the Candida species . Candidacidal activity of free and liposomal amphotericin B resulted in comparable effects; however, the killing of the yeasts by liposomal amphotericin B was slower than by free amphotericin B.

Antimicrob Agents Chemother, 1995 Apr, 39(4), 924 - 9
Fluconazole, D0870, and flucytosine treatment of disseminated Candida tropicalis infections in mice; Graybill JR et al.; D0870 is a recently developed triazole with characteristics of a broad spectrum of activity and slow clearance by nonrenal mechanisms . Herein we have evaluated the efficacy of D0870, alone and combined with flucytosine, in a murine model of disseminated Candida tropicalis infection . Four isolates of C . tropicalis were evaluated . Two were highly susceptible in vitro to fluconazole, and two were resistant to fluconazole . All were highly susceptible to flucytosine and D0870 . Animals were pretreated with 5-fluorouracil 1 day before infection because C . tropicalis has reduced virulence in immunocompetent mice . This was done to render them neutropenic for > 10 days . Mice were infected intravenously and treated orally with D0870 or fluconazole, alone or combined with flucytosine . Survival and tissue burden of the spleen and kidneys were used to evaluate the efficacy of antifungal therapy . Fluconazole was less effective for treatment of resistant C . tropicalis than susceptible C . tropicalis . D0870 was more potent than fluconazole and was effective in fluconazole-resistant isolates . Flucytosine was consistently effective when used alone but did not consistently add to the benefit of D0870 or fluconazole . D0870 has potential in treatment of candidiasis caused by C . tropicalis, including fluconazole-resistant isolates.

Appl Environ Microbiol, 1995 Apr, 61(4), 1420 - 5
Phosphorus-31 and carbon-13 nuclear magnetic resonance study of glucose and xylose metabolism in agarose-immobilized Candida tropicalis; Lohmeier-Vogel EM et al.; Candida tropicalis can ferment both hexose and pentose sugars . Here, we have used 31P and 13C nuclear magnetic resonance spectroscopy to study the capacity of this yeast species to metabolize glucose or xylose when immobilized in small (< 1-mm-diameter) agarose beads . Immobilized C . tropicalis metabolizing glucose showed rapid initial growth within the beads . A corresponding drop in the intracellular pH (from 7.8 to 7.25) and hydrolysis of intracellular polyphosphate stores were observed . Although the initial rate of glucose metabolism with immobilized C . tropicalis was similar to the rate observed previously in cell suspensions, a decrease by a factor of 2.5 occurred over 24 h . In addition to ethanol, a significant amount of glycerol was also produced . When immobilized C . tropicalis consumed xylose, cell growth within the beads was minimal . The intracellular pH dropped rapidly by 1.05 pH units to 6.4 . Intracellular ATP levels were lower and intracellular Pi levels were higher than observed with glucose-perfused cells . Consumption of xylose by immobilized C . tropicalis was slower than was previously observed for oxygen-limited cell suspensions, and xylitol was the only fermentation product.

Appl Environ Microbiol, 1995 Apr, 61(4), 1414 - 9
Phosphorus-31 and carbon-13 nuclear magnetic resonance studies of glucose and xylose metabolism in Candida tropicalis cell suspensions; Lohmeier-Vogel EM et al.; The metabolism of glucose and xylose was studied as a function of oxygenation in suspensions of Candida tropicalis by 31P and 13C nuclear magnetic resonance spectroscopy . Both the rate of carbohydrate metabolism and the cytoplasmic pH were independent of the rate of oxygenation in cells metabolizing glucose . However, these two parameters were markedly dependent on the rate of oxygenation in C . tropicalis cells metabolizing xylose . For example, the cytoplasmic pH in fully oxygenated xylose-metabolizing cells was 7.8 but decreased to 6.3 in anoxic cells . In general, suspensions of cells consuming xylose had a lower rate of sugar uptake, a more acidic cytoplasmic pH, lower levels of sugarphosphomonoesters (SP) and ATP, higher levels of intracellular Pi, a more alkaline vacuolar pH, and a lower rate of extracellular Pi assimilation and polyphosphate synthesis than cells consuming glucose . These observations indicate that C . tropicalis metabolizing xylose is less energized than glucose-metabolizing cells . On both carbon sources, however, an inverse correlation between intracellular levels of SP and Pi was observed . Also, uptake of extracellular Pi correlated with the synthesis of polyphosphates within the cells . During anoxia, Pi was not taken up, and polyphosphates were hydrolyzed instead to fulfill the cells' requirements for phosphate.

South Med J, 1995 Apr, 88(4), 465 - 6
Candidemia after endoscopic retrograde cholangiopancreatography; Ojeas H et al.; We report a case of candidemia associated with manipulation of the common bile duct during endoscopic retrograde cholangiopancreatography . The patient had received parenteral nutrition and prolonged antibiotic therapy before the procedure and before Candida tropicalis was grown from blood cultures . Antifungal therapy resulted in clinical response.

Eur J Clin Microbiol Infect Dis, 1995 Apr, 14(4), 362 - 5
In vitro activity of amphotericin B, flucytosine and fluconazole against yeasts causing bloodstream infections; Berenguer J et al.; The in vitro activity of amphotericin B, flucytosine and fluconazole against 95 yeasts causing fungemia in a single institution over the last eight years was determined by a broth macromethod recommended by the National Committee for Clinical Laboratory Standards . All strains were inhibited by amphotericin B concentrations of < or = 1 microgram/ml . With flucytosine in most species the MIC50 was between 0.12 and 0.25 microgram/ml and the MIC90 was between 0.25 and 1 microgram/ml . One exception with flucytosine was Candida krusei, with an MIC50 and MIC90 of 16 micrograms/ml and 32 micrograms/ml, respectively . Overall, 12% of the isolates needed at least 8 micrograms/ml of fluconazole to be inhibited . Fluconazole was very active against Candida albicans, Candida tropicalis and Cryptococcus neoformans, with MIC50 ranging from 0.12 to 0.5 microgram/ml and MIC90 of 1 microgram/ml, and somewhat less active against Candida parapsilosis (MIC50 of 1 microgram/ml and MIC90 of 4 micrograms/ml) . Fluconazole exhibited poor in vitro activity against Candida krusei (MIC50 and MIC90 of 64 micrograms/ml) and Torulopsis glabrata (MIC50 of 4 micrograms/ml and MIC90 of 16 micrograms/ml) . High MICs of fluconazole were found for four strains of Candida albicans, one with an MIC of 4 micrograms/ml and three (5.7%) with MICs of > or = 16 micrograms/ml . Previous exposure to fluconazole could be demonstrated in two of these strains . Further work must be done in order to determine appropriate breakpoints of antifungal agents, to assess the clinical relevance of azole resistance in yeasts causing bloodstream infections and to identify risk factors for infections with azole-resistant yeasts.

J Leukoc Biol, 1995 Apr, 57(4), 651 - 6
Effects of granulocyte colony-stimulating factor and interferon-gamma on antifungal activity of human polymorphonuclear neutrophils against pseudohyphae of different medically important Candida species; Roilides E et al.; Polymorphonuclear neutrophils (PMNs) are the major host defense against pseudohyphae, the invasive form of Candida species . We studied the effects of granulocyte colony-stimulating factor (G-CSF) and interferon-gamma (IFN-gamma) on the PMN-induced damage of pseudo-hyphae of Candida albicans, Candida tropicalis, and Candida parapsilosis in vitro by using two antifungal assays: a modified limiting dilution assay and a colorimetric metabolic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay . PMNs from healthy volunteers were incubated with either G-CSF (100-10,000 U/ml) or IFN-gamma (10-5000 U/ml) or buffer at 37 degrees C for 90 min and their capacity to damage nonopsonized pseudohyphae was then measured . C . tropicalis appeared to be the most susceptible species, whereas C . parapsilosis showed the highest rate of resistance to PMN damage . G-CSF (500-10,000 U/ml) and IFN-gamma (100-1000 U/ml) enhanced the antifungal activity of PMNs against C . albicans pseudo-hyphae (P < .01 and P < .05) . Among the others, G-CSF enhanced PMN-induced damage of C . parapsilosis at concentrations 500-10,000 U/ml (P < .05), whereas it enhanced damage of C . tropicalis only at 10,000 U/ml (P < .01) . IFN-gamma (100-1000 U/ml)-primed PMNs also caused augmented damage of C . parapsilosis (P < .05) but not of C . tropicalis at the same concentrations . Species-dependent differences exist in the responses of PMNs to Candida pseudohyphae and G-CSF as well as IFN-gamma are important immunomodulators of phagocytic host defenses against them.

Gene, 1995 Mar 21, 155(1), 123 - 8
Isolation, characterization and expression of the gene that encodes D-arabinitol dehydrogenase in Candida tropicalis; Murray JS et al.; The gene (ARD) that encodes NAD-dependent D-arabinitol dehydrogenase (ArDH) in the pathogenic fungus Candida tropicalis (Ct) was cloned by transforming Escherichia coli (Ec) BW31M (araCc) with a plasmid library of Ct genomic DNA and selecting for D-arabinitol-utilizing (D-arab+) clones . Plasmid DNA from a D-arab+ clone retransformed fresh Ec BW31M cells to D-arab+; these cells produced both ArDH catalytic activity and a 31-kDa protein recognized by antibodies to native Ct ArDH . The plasmid contained an 846-bp open reading frame (ORF) that encoded a deduced protein of 282 amino acids (aa) (30,748 Da) . Four partial aa sequences from Ct ArDH were present in the deduced aa sequence, thus verifying that Ct ARD had been cloned . Ct ArDH was 95% identical to ArDH from Candida albicans (Ca), 85% identical to a xylitol dehydrogenase (XDH) from Pichia stipitis (Ps) and 20-25% identical to many other short-chain dehydrogenases . Ct ArDH, Ca ArDH and Ps XDH were typical short-chain dehydrogenases except that they lacked an N-terminal Gly that is conserved in other members of this family . Thus, these enzymes may represent a subclass of closely-related fungal pentitol dehydrogenases . Large amounts of recombinant ArDH (re-ArDH) were produced in Ec and purified by dye ligand affinity chromatography . The physical and catalytic properties of re-ArDH were similar to those of native Ct ArDH, and re-ArDH and native ArDH performed similarly in an automated enzymatic assay for D-arabinitol in human serum.

Am J Obstet Gynecol, 1995 Mar, 172(3), 1045 - 7
Candida tropicalis chorioamnionitis; Nichols A et al.; A case of Candida tropicalis chorioamnionitis at 25 weeks' gestation is presented, and the literature is reviewed . This is the first report, to our knowledge, of C . tropicalis diagnosed antenatally with confirmed congenital infection.

Infect Immun, 1995 Mar, 63(3), 976 - 83
Penetration and damage of endothelial cells by Candida albicans; Filler SG et al.; The mechanisms of phagocytosis of Candida albicans by human vascular endothelial cells and subsequent endothelial cell injury were examined in vitro . Both live and killed C . albicans cells were phagocytized by endothelial cells . This organism specifically induced endothelial cell phagocytosis because neither Candida tropicalis nor Torulopsis glabrata was ingested . Endothelial cell microfilaments polymerized around C . albicans as the organisms were phagocytized . Cytochalasin D inhibited this polymerization of microfilaments around C . albicans and blocked phagocytosis . The blocking of actin depolymerization with phalloidin had no effect on microfilament condensation around the organism, indicating that the microfilaments surrounding C . albicans are formed from a pool of G-actin . Intact microtubules were also necessary for the phagocytosis of C . albicans, since the depolymerizing of endothelial cell microtubules with nocodazole prevented the condensation of actin filaments around the organisms and inhibited phagocytosis . In contrast, microtubule depolymerization was not required for microfilament function because the blocking of microtubule depolymerization with taxol had no effect on microfilament condensation around C . albicans . The phagocytosis of C . albicans was pivotal in the induction of endothelial cell damage, since the blocking of candidal internalization significantly reduced endothelial cell injury . Endothelial cells were not damaged by phagocytosis of dead organisms, indicating that injury was caused by a factor associated with viable organisms . Therefore, C . albicans is uniquely able to induce endothelial cell phagocytosis by comparison with non-albicans species of Candida . Furthermore, at least two components of the endothelial cytoskeleton, microfilaments and microtubules, are necessary for the phagocytosis of C . albicans.

Clin Infect Dis, 1995 Mar, 20(3), 571 - 5
Candidemia in a pediatric population; Stamos JK et al.; Candidemia results in a mortality of > 50% among adults, but data on children with candidemia are limited . We reviewed 70 episodes of pediatric candidemia that occurred between January 1988 and October 1992 . Of these episodes, 53% were caused by Candida albicans, 24% were caused by Candida parapsilosis, 16% were caused by Candida tropicalis, and 3% were caused by Candida krusei . Twenty-five percent of the patients were premature infants . Other underlying conditions included malignancy (15%); cardiac disease (14%); and short-gut syndrome (14%) . A central venous catheter was in place during 61 (87%) of 70 episodes . Candiduria preceded candidemia in only 4 (8%) of 52 patients . The overall mortality rate was 19%; 36% of those with intravenous catheters that were not removed within 3 days died, whereas none of the patients from whom catheters were removed within 3 days died (P < .0001) . Only two survivors had complications . Therapy with amphotericin B (with or without flucytosine) was administered to 74% of these patients . Seventeen patients were not treated medically; all were immunocompetent and survived . Of these patients, 15 were > 2 months of age; 14 had candidemia for < or = 2 days; and 15 had an intravenous catheter removed within 2 days of the onset of candidemia . No patient stopped receiving amphotericin B because of side effects . The results of this study suggest the following: that mortality associated with candidemia is lower among children than among adults; that failure to remove the indwelling intravenous catheter usually results in a poor outcome; that candiduria rarely precedes candidemia in children; and that amphotericin B is well tolerated by children.

J Clin Microbiol, 1995 Mar, 33(3), 535 - 40
Comparison of Etest and National Committee for Clinical Laboratory Standards broth macrodilution method for azole antifungal susceptibility testing; Colombo AL et al.; The use of Etest strips for antimicrobial susceptibility testing is a new and promising method with broad applications in microbiology . Since these strips contain a predefined continuous gradient of a drug, it is possible to obtain a reproducible, quantitative MIC reading . We performed a prospective and double-blinded study to compare the Etest and National Committee for Clinical Laboratory Standards (Villanova, Pa.) broth macrodilution methods for determining the MICs of fluconazole, itraconazole, and ketoconazole for 100 clinical isolates (25 Candida albicans, 25 Cryptococcus neoformans var . neoformans, 20 Torulopsis {Candida} glabrata, 15 Candida tropicalis, and 15 Candida parapsilosis) . The Etest method was performed according to the manufacturer's instructions, and the reference method was performed according to National Committee for Clinical Laboratory Standards document M27-P guidelines . Despite differences between results for some species-drug combinations, Etest and macrobroth MICs were, in general, in good agreement . The MIC agreement rates for the two methods, within +/- 1 dilution, were 71% for ketoconazole, 80% for fluconazole, and 84% for itraconazole . According to our data, Etest has potential utility as an alternative method.

Bratisl Lek Listy, 1995 Mar, 96(3), 168 - 72
{Results of mycologic examinations of sputum in patients with pulmonary diseases during a 10-year period (1984-1993)}; Kockovska D; The study of present results of mycologic examinations of samples of sputums taken during the period from 1984 to 1993 from patients with pulmonary diseases . The examined group was constituted of 2,452 subjects, out of which 2/3 were male and 1/3 were female patients . The risk age category was discovered to be that over 61 years of age (32%) . 70% of mycologic cultivations were positive . 1.640 etiologic agents were isolated from the positive samples . The most frequently isolated yeast species were Candida albicans (61.7%), Candida tropicalis (8.2%), Torulopsis sp . (4.9%), the most frequent fibrous microscopic fungi findings included Penicillium sp . (4.5%) and Aspergillus sp . (2.9%) . The most frequent diagnoses included bronchitis ac . chr . 15%), pneumonia 13%, TBC of the lungs 11.7%, asthma bronchiale 10.6%, and bronchopneumonia 7% . Repeatedly C . albicans was the most frequently isolated species, namely in the range 52-80%.

Arch Pharm (Weinheim), 1995 Feb, 328(2), 197 - 201
Cytotoxicity and antimicrobial activity of some naphthol derivatives; Shen AY et al.; 2-Hydroxymethyl-1-naphthol diacetate (TAC) and sixteen Mannich base derivatives of naphthol were prepared and examined for cytotoxicity and antimicrobial activity . Cytotoxicity was examined against four human carcinoma cell lines . Several derivatives were effective at concentrations < 4 micrograms/ml . TAC showed the highest cytotoxicity . Inhibition of DNA-, RNA-, and protein synthesis by TAC was also studied and discussed . TAC also exhibits potent antimicrobial activity against Enterobacter clocae 23355, Klebsiella pneumonia 13883, Proteus vulgaris 13315, Pseudomonas aeruginosa 27853, Candida parapsilosis, Candida tropicalis, Trichosposon beigelli, and Rhodotorul spp . with minimum inhibitory concentrations of 0.1-0.4 microM . These results indicate that esterification by Bruson reaction of 1-naphthol Mannich base to TAC enhances the cytotoxicity and antimicrobial activity.

Arch Microbiol, 1995 Feb, 163(2), 104 - 11
Novel NADP-linked isocitrate dehydrogenase present in peroxisomes of n-alkane-utilizing yeast, Candida tropicalis: comparison with mitochondrial NAD-linked isocitrate dehydrogenase; Yamamoto S et al.; Peroxisomal NADP-linked isocitrate dehydrogenase (Ps-NADP-IDH) was purified for the first time from Candida tropicalis cells grown on n-alkane as a carbon source, which was effective in proliferation of peroxisomes . The properties of Ps-NADP-IDH were compared with those of mitochondrial NAD-linked isocitrate dehydrogenase (Mt-NAD-IDH) purified from the cells grown on acetate, in which peroxisomes did not proliferate . Ps-NADP-IDH was a homodimer of identical subunits (45 kDa), while Mt-NAD-IDH was suggested to be a heterooctamer composed of two types of subunits with different molecular masses (41 and 38 kDa) . Kinetic studies revealed that Ps-NADP-IDH gave Michaelis-Menten saturation curves against isocitrate and NADP concentrations, whereas Mt-NAD-IDH was an allosteric enzyme regulated by ATP, AMP, and citrate . Inhibition by 2-oxoglutarate, a precursor of glutamate, was observed only for Ps-NADP-IDH . Both enzymes were inhibited by concomitant addition of oxalacetate and glyoxylate . The function of Ps-NADP-IDH seems to be completely discriminated from that of Mt-NAD-IDH as reflected by their distinct subcellular localizations . Furthermore, the properties of Ps-NADP-IDH were also compared with those of other mitochondrial and cytosolic IDHs from sources reported previously.

Klin Med (Mosk), 1995, 73(3), 44 - 7
{The clinical picture of mycotic complications in HIV-infected patients}; Rakhmanova AG et al.; Mycotic complications were registered in 21 out of 37 HIV-infected subjects . Oropharyngeal candidiasis was most common . It occurred prior to or concurrently with esophageal and skin candidiasis, fungemia, meningoencephalitis and disseminated lesions . With immunodeficiency progression, the prevalence and severity of mycosis go up . The causing fungi vary in great range: Candida albicans, Candida krusei . Candida tropicalis, Candida pseudotropicalis, Candida parapsilosis . Cryptococcus neoformans, Rhodotorula rubra, Penicillium chrysogenum.

Chemotherapy, 1995 Jan-Feb, 41(1), 39 - 44
Candida glabrata, Candida krusei, non-albicans Candida spp., and other fungal organisms in a sixty-bed national cancer center in 1989-1993: no association with the use of fluconazole; Kunova A et al.; During the 5-year period 1989-1993, the incidence of Candida krusei, and other non-albicans Candida spp., was analyzed in a 60-bed cancer department . The frequency of C . krusei, before fluconazole was introduced into therapeutic protocols in 1990, was 16.5%, and after introduction of fluconazole into prophylaxis in acute leukemia in 1991, the incidence of C . krusei was 12.7% . After 3 years of using this drug in therapy and prophylaxis, the incidence of C . krusei in 1993 was 14.8%, what was lower than before this drug was introduced in our country . 97.6% of all isolated fungi were yeasts and only 2.4% were molds . Among yeasts, the most frequently isolated pathogen was Candida albicans with 64.3% in 1989 and 74.2% in 1993 . The next was C . krusei with 21.2% in 1992 and 16.5% in 1989, but 14.8% in 1993, and Candida tropicalis and Candida glabrata with 9.03% in 1989 and 2.7% in 1993 . Among the molds, Aspergillus spp . was the most frequently isolated genus . Analyzing the etiology of mycologically proven fungal infections confirmed by positive blood cultures or biopsies, C . albicans and Aspergillus spp . were the most common causative organisms.

Clin Infect Dis, 1995 Jan, 20(1), 115 - 25
Importance of Candida species other than C . albicans as pathogens in oncology patients; Wingard JR; A number of surveys have documented increased rates of candida infection over the past several decades . In this assessment of the frequency and distribution of non-albicans Candida species among patients with cancer, 37 reports that were published between 1952 and 1992 and that described 1,591 cases of systemic candida infection were reviewed . Species other than Candida albicans accounted for 46% of all systemic candida infections in patients with cancer; specifically, Candida tropicalis accounted for 25%, Candida glabrata for 8%, Candida parapsilosis for 7%, and Candida krusei for 4% . Other species were uncommon . C . tropicalis was the predominant pathogenic Candida species in five reports, C . glabrata in two, C . krusei in two, and Candida stellatoidea in one . The perception that, over time, a greater proportion of candida infections have been caused by non-albicans species was not borne out . The wide variability in reported findings was striking and was due in part to differences in the underlying disease affecting the patients described . For example, patients with leukemia were more likely to be infected by C . albicans or C . tropicalis but less likely to be infected by C . glabrata than patients with other types of cancer . The recent increase in the rate of bone marrow transplantation may also have contributed to discrepancies among reports . Bone marrow transplant recipients were more likely to be infected by C . krusei or C . lusitaniae . The other factors partially responsible for the variability among reports included common-source contamination and the pressures imposed by antimicrobial measures.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 27 - 31
Hyaluronidase and chondroitin sulphatase production by different species of Candida; Shimizu MT et al.; The production of hyaluronidase and chondroitin sulphatase by Candida albicans, Candida tropicalis, Candida parapsilosis, Candida guilliermondii and Candida krusei was investigated using a complex culture medium (Sabouraud glucose agar) and a chemically defined medium . Among the 63 C . albicans isolates tested, 61 (97.8%) were found to be hyaluronidase and chondroitin sulphatase producers; one isolate produced only chondroitin sulphatase and one other was unable to produce either enzyme . The second major hyaluronidase and chondroitin sulphatase producing species was C . tropicalis followed by C . guilliermondii, C . parapsilosis and C . krusei . Among the C . albicans isolates tested no relation between the source of isolation and the amount of hyaluronidase and chondroitin sulphatase produced was found.

Nat Toxins, 1995, 3(3), 138 - 44
Metabolism of the Fusarium mycotoxins zearalenone and deoxynivalenol by yeast strains of technological relevance; Boswald C et al.; The Fusarium mycotoxin zearalenone (ZEA), added at a level of 2 micrograms/ml, was reduced stereoselectively by cultures of Candida tropicalis, Torulaspora delbruckii, Zygosaccharomyces rouxii, and 7 Saccharomyces strains to both alpha- and beta-zearalenol . In contrast, only alpha-zearalenol was produced from ZEA by Pichia fermentans and several yeast strains of the genera Candida, Hansenula, Brettanomyces, Schizosaccharomyces, and Saccharomycopsis . No glucose conjugates of ZEA (zearalenone-4-beta-D-glucopyranoside) were detected . The trichothecene mycotoxin deoxynivalenol (DON) was not metabolized by any of the yeast strains that were used for analysis.

Acta Haematol, 1995, 94(3), 148 - 51
Diagnosis of disseminated candidiasis by fine needle aspiration of lymph node and by splenic imprint in a patient with acute promyelocytic leukemia; Chao TY et al.; Cytologic studies were done on fine needle aspirates of the lymph node and imprints of splenic biopsies from a patient with acute promyelocytic leukemia who was febrile while being treated with chemotherapy . Examination of the lymph node aspirates revealed pus and numerous pseudohyphae which were later identified as Candida tropicalis . When multiple nodular lesions were detected in the spleen by abdominal sonography and CT scan, needle biopsy of the spleen was done . Cytologic examination of touch imprints of the biopsy disclosed intracellular fungal blastospores . The patient was treated with and responded well to amphotericin B and 5-fluorocytosine . As a result of our experience with this patient we emphasize the importance of close incorporation of clinical information and diagnostic cytology . With such a cooperation, cytologic studies become a most useful method for diagnosis.

Clin Infect Dis, 1994 Dec, 19(6), 1049 - 53
Fungal infection of ventriculoperitoneal shunts in children; Chiou CC et al.; Infection is still the most common complication of shunt procedures in children . However, fungal infection is still considered to be rare . We found that fungi accounted for 17% of shunt infections (8 of 48) in a retrospective study . All of the patients were premature babies and had received a ventriculoperitoneal shunt because of hydrocephalus . The clinical manifestations were subtle and insidious . The time of onset of infection ranged from 1 month to 1 year after the insertion of the shunt . Examination of the cerebrospinal fluid of infected patients showed mild pleocytosis with an elevated protein concentration . Candida species (including Candida albicans, Candida parapsilosis, and Candida tropicalis) or Torulopsis glabrata were isolated . In all but one case, shunts were removed and systemic therapy with amphotericin B was administered . Amphotericin B was given intrathecally to two patients, who did not respond to systemic therapy . Treatment with fluconazole failed for one patient . We suggest performing fungal cultures in cases of shunt infection, especially those involving premature infants . Extraventricular drainage, systemic therapy with amphotericin B, and insertion of a new shunt remain the principal components of the treatment regimen for fungal shunt infections in children.

J Clin Microbiol, 1994 Dec, 32(12), 3034 - 6
Rapid identification of Candida albicans by using Albicans ID and fluoroplate agar plates; Rousselle P et al.; Two commercially available agar media, Albicans ID and Fluoroplate, that use a chromogenic or a fluorogenic substrate for the detection and identification of Candida albicans were evaluated . From 1,006 clinical samples containing 723 yeast strains, 352 C . albicans strains were detected with either of the two media . The sensitivity of each of the two media was 93.8% and the specificity was 98.6%, with five false-positive reactions for Candida tropicalis and no false-negative reactions.

J Clin Microbiol, 1994 Dec, 32(12), 2889 - 92
Misidentification of clinical yeast isolates by using the updated Vitek Yeast Biochemical Card; Dooley DP et al.; The Vitek Yeast Biochemical Card (YBC) is widely used as a rapid identification (RI) (within 48 h) system for clinical yeast isolates . We compared the RI results obtained by the YBC technique with matched results obtained with the API 20C system . The RI of germ tube-negative yeasts isolated from 222 clinical specimens was performed with the YBC system, and the results were compared with those of standard identifications obtained by using the API 20C system and morphology, with additional biochemical reactions performed as required . Commonly isolated yeasts (Candida albicans {n = 29}, Candida tropicalis {n = 40}, Torulopsis {Candida} glabrata {n = 28}, Candida parapsilosis {n = 12}, and Cryptococcus neoformans {n = 14}) were generally well identified (115 of 123 {93%} identified correctly, with only C . albicans, C . tropicalis, and C . neoformans mis- or unidentified more than once) . The RI of less commonly isolated yeasts included in the YBC database, however, was less successful (54 of 99 {55%} correct) . The YBC card failed to identify 42% (10 of 24) of Candida krusei isolates, 80% (4 of 5) of Candida lambica isolates, 88% (7 of 8) of Trichosporon beigelii isolates, and 83% (10 of 12) of Cryptococcus isolates (non-C . neoformans species) . For most identification failures (79%; 42 of 53) there was no identification by the end of 48 h; the other identification failures (21%; 11 of 53) gave definite but incorrect identifications . Of eight rare clinical yeast isolates not included in the Vitek database, six were correctly, not identified, while two (25%) were falsely assigned a definite RI (one Hansenula fabianii isolate was identified as Rhodotorula glutinis, and one Hansenula isolate {non-Hansenula anomala} was identified as Hansenula anomala) . While the Vitek YBC rapidly and adequately identifies common yeast isolates, it fails in the RI of more unusual organisms.

Plant Foods Hum Nutr, 1994 Dec, 46(4), 345 - 51
Utilization of cassava peels as substrate for crude protein formation; Antai SP et al.; Mash prepared from cassava peels was inoculated with either Sacchromyces cerevisiae or Candida tropicalis and then left to ferment for 7 days . Chemical analysis of the fermented mash showed substantial increase in crude protein content and decrease in carbohydrate content of the mash . The results also revealed slight increases in the ash, fibre and lipid content of the fermented mash . It was further observed that, when the mash was supplemented with inorganic nitrogen sources (urea, ammonium sulfate or sodium nitrate) before commencement of fermentation, the amount of crude protein formed was almost doubled . This enhanced crude protein production was highest in the mash supplemented with urea . Temperature of 30 degrees C, pH of 5.5 and moisture concentration of 130% were found to be optimum for crude protein formation by the organisms growing on the mash.

Bone Marrow Transplant, 1994 Dec, 14(6), 919 - 24
Antifungal prophylaxis with low-dose fluconazole during bone marrow transplantation . The Bone Marrow Transplantation Team; Alangaden G et al.; The present study investigated the prophylactic efficacy of fluconazole at 100-200 mg/day against invasive fungal infections during bone marrow transplantation (BMT) . During July 1990 to December 1991, all BMT recipients received antifungal prophylaxis with fluconazole at either 200 mg/day or 100 mg/day . Historical controls were those that received no antifungal prophylaxis (January 1989 to June 1990) . Fungemia occurred in 4 of 112 fluconazole recipients and 8 of 79 controls (p < 0.05) prior to engraftment . Torulopsis (Candida) glabrata (three patients), Cryptococcus terreus and Candida tropicalis (mixed in one patient) caused fungemia in four patients in the fluconazole group; Candida albicans caused six of eight fungemic episodes in the controls . All three Torulopsis glabrata isolates were fluconazole-resistant . Colonization due to C . albicans was markedly diminished in the fluconazole group (19 of 112 patients versus 53 of 79 controls) . T . glabrata, on the other hand, was a more common colonizing organism in the fluconazole group (36 of 112 vs 10 of 79) . The frequency of isolating C . albicans and/or T . glabrata was significantly different between fluconazole and control groups (p < 0.0001) . Empiric use of amphotericin B therapy was markedly reduced in the fluconazole group (4.5% vs 34%; p < 0.0001) . Fluconazole at 200 mg/day or 100 mg/day appeared equally effective . Fluconazole at a daily dose of 100 mg or 200 mg as antifungal prophylaxis during BMT: (1) significantly reduced the frequency of systemic fungal infections, (2) markedly reduced colonization and infection due to C . albicans, and (3) markedly reduced the need for empiric amphotericin B therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

Med Clin (Barc), 1994 Nov 5, 103(15), 579 - 82
{Sepsis by Candida tropicalis in patients with granulocytopenia . A study of 10 cases}; Terol MJ et al.; The aim of the present study was to analyze the main clinical and evolutive characteristics of a series of 10 patients diagnosed with sepsis by Candida tropicalis over a 5-year period in a Hematology Unit . The mean age of the 10 patients was 23 years (range 13-66 years) with 6 males and 4 females . Eight patients had acute leukemia, 1 non-Hodgkin's lymphoma and another patient had severe bone marrow aplasia . All the patients presented intense granulocytopenia (< 0.5 x 10(9)/L), had intravenous catheters and were receiving wide spectrum antibiotics as treatment for bacterial infection . The diagnosis of the fungal infection was based on the growth of C . tropicalis in blood cultures together with the evidence of tissue involvement by the fungus . Fever (> 38 degrees C) was the initial symptom of the infection in all the patients, being accompanied by myalgia in 5 cases, pleuritic pain in 2 and septic shock in 1 . Violaceous erthymatomous pustules disseminated over the trunk and limbs, the histologic study of which demonstrated the presence of C . tropicalis were observed in 9 patients . Septic metastasis were found in the liver (2 cases), serosae (2 cases), the psoas muscle and the brain (1 case), respectively . Eight patients underwent treatment with amphotericin B which was complemented with 5-fluorocytosin in 6, with death occurring in the remaining 2 patients prior to the start of treatment . Three patients died with active fungal infection (2 by cerebral hemorrhage and 1 by septic shock) . In 2 patients the infection evolved to chronic systemic candidiasis and in the remaining 5 patients infection was resolved with hemoperipheral values returning to normal . Sepsis by Candida tropicalis is a severe complication in patients with granulocytopenia, being mainly characterized by fever, cutaneous papulae and, to a lesser extent, muscle pain . Amphotericin B alone, or in combination with 5-fluorocytosin constitute a treatment of choice in this infection, which nonetheless is associated with an undisdainful mortality.

Yeast, 1994 Nov, 10(11), 1467 - 76
Near-stoichiometric interaction between the non-specific lipid-transfer protein of the yeast Candida tropicalis and peroxisomal acyl-coenzyme A oxidase prevents the thermal denaturation of the enzyme in vitro; Niki T et al.; A 14-kDa peroxisomal-matrix protein, named PXP-18, of the yeast Candida tropicalis is a structural and functional homologue of the mammalian nonspecific lipid-transfer protein (identical to sterol carrier protein-2) . PXP-18 protected acyl-coenzyme A oxidase (ACO), the rate limiting enzyme of the peroxisomal beta-oxidation of fatty acids, from thermal inactivation at 48 degrees C or 70 degrees C . This effect was dose-dependent and not replaceable either by chicken egg white lysozyme, which is similar to PXP-18 (insofar as it is basic, small, and monomeric), or by bovine serum albumin, a carrier of lipids in the blood . ACO was irreversibly denatured by heat treatment at 70 degrees C for 15 min . However, when ACO and PXP-18 were similarly heat-treated, they formed a large complex at a molar ratio of PXP-18 to ACO subunit that was about one, independent of their initial ratio . This near-stoichiometric complex had ACO activity after a 500-fold dilution and was accompanied by ACO that was free of PXP-18 and indistinguishable from native ACO in size and activity . PXP-18 also protected urate oxidase, another peroxisomal enzyme, from inactivation at 66 degrees C for 15 min and facilitated the renaturation of ACO denatured by 2 M urea . These results indicated that PXP-18 is active in modulating the structure of peroxisomal enzymes in vitro . It is possible that PXP-18 functions as a stress protein or as a part of the system that keeps peroxisomal proteins intact.

J Clin Microbiol, 1994 Oct, 32(10), 2494 - 500
Comparative study of broth macrodilution and microdilution techniques for in vitro antifungal susceptibility testing of yeasts by using the National Committee for Clinical Laboratory Standards' proposed standard; Barchiesi F et al.; A comparative study of broth macro- and microdilution methods for susceptibility testing of fluconazole, itraconazole, flucytosine, and amphotericin B was conducted with 273 yeasts . The clinical isolates included 100 Candida albicans, 28 Candida tropicalis, 25 Candida parapsilosis, 15 Candida lusitaniae, 15 Candida krusei, 50 Cryptococcus neoformans var . neoformans, 25 Torulopsis (Candida) glabrata, and 15 Trichosporon beigelii strains . Both methods were performed according to the National Committee for Clinical Laboratory Standards' (NCCLS) recommendations (document M27-P) . For fluconazole, itraconazole, and flucytosine, the endpoint was the tube that showed 80% growth inhibition compared with the growth control for the macrodilution method and the well with slightly hazy turbidity (score 1) compared with the growth control for the microdilution method . For amphotericin B, the endpoint was the tube and/or well in which there was absence of growth . For the reference macrodilution method, the MICs were determined after 48 h of incubation for Candida spp., T . glabrata, and T . beigelii and after 72 h for C . neoformans var . neoformans . For the microdilution method, either the first-day MICs (24 h for all isolates other than C . neoformans and 48 h for C . neoformans var . neoformans) or the second-day MICs (48 and 72 h, respectively) were evaluated . The agreement within one doubling dilution of the macrodilution reference for all drugs was higher with the second-day MICs than with the first-day MICs for the microdilution test for most of the tested strains . General agreement was 92% for fluconazole, 85.7% for itraconazole, 98.3% for flucytosine, and 96.4% for amphotericin B . For C . neoformans var . neoformans and T . beigelii, the agreement of the first-day reading was higher than that of the second-day reading for fluconazole (94 versus 92%, respectively, for C . neoformans var . neoformans, and 86.7 versus 80%, respectively, for T . beigelii) . Our studies indicate that the microdilution technique performed following the NCCLS guidelines with a second-day reading is a valid alternative method for testing fluconazole, itraconazole, flucytosine, and amphotericin B against these eight species of yeasts.

Clin Infect Dis, 1994 Oct, 19(4), 697 - 703
Candida tropicalis vertebral osteomyelitis: a late sequela of fungemia; Ferra C et al.; Two adult patients who had undergone bone marrow transplantation had transient fungemia due to Candida tropicalis during the posttransplantation neutropenic period and later (at 5 and 14 months post-transplantation) developed vertebral osteomyelitis due to the same organism . The courses of all adult patients who underwent bone marrow transplantation at our center during this time were reviewed for determining the frequency of this problem . Molecular typing techniques were used to establish the relationship between the organisms isolated during the initial fungemia and those causing the subsequent osteomyelitis . Only two of 532 adults who received transplants at our center from 1980 to 1993 developed candidal osteomyelitis . Moreover, they are part of a subset of 13 patients (2.4% of the total) whose cultures were positive for C . tropicalis; five of the 13 had fungemia . The study of fungal isolates from prior sites of colonization and from blood sampled during the original fungemia and of subsequently recovered vertebral bone isolates by means of DNA restriction fragment analysis (with use of standard agarose gel electrophoresis or pulsed-field gel electrophoresis) showed that the colonizing, bloodstream, and bone isolates were identical in each case . Transient C . tropicalis fungemia in severely immunocompromised patients may cause important late infectious complications, including osteomyelitis . Although these initial septic events may appear to resolve easily, the outcome in the two cases in this report suggests that special treatment considerations, such as immediate removal of the central venous catheter and intensive treatment with amphotericin B, may be required in selected cases for prevention of late sequelae.

Mol Cell Endocrinol, 1994 Sep, 104(2), 127 - 31
The sequence of porcine 80 kDa 17 beta-estradiol dehydrogenase reveals similarities to the short chain alcohol dehydrogenase family, to actin binding motifs and to sterol carrier protein 2; Leenders F et al.; The cDNA of porcine 17 beta-estradiol dehydrogenase codes for a polypeptide of 737 amino acids . The dehydrogenase activity of the 80 kDa translation product is located in its N-terminal 32 kDa fragment, which is the major form isolated from endometrial epithelium . beta-Actin co-purifies with some of the 32 kDa enzyme, which contains actin-binding motifs and is homologous to hydratase-dehydrogenase-epimerase of Candida tropicalis . The microbody-targeting signal AKI and sequences resembling sterol carrier protein 2 are present in the C-terminal part of the 80 kDa protein . The N- and C-terminal parts are connected by a sequence containing the putative protease recognition signal AAP.

Eur J Clin Invest, 1994 Sep, 24(9), 586 - 99
Host defence capacities of pulmonary surfactant: evidence for 'non-surfactant' functions of the surfactant system; Pison U et al.; The most well characterized function of pulmonary surfactant is its ability to reduce surface tension at the alveolar air-liquid interface, thereby preventing lung collapse . However, several lines of evidence suggest that surfactant may also have 'non-surfactant' functions: specific components of surfactant (proteins and phospholipids) may interact with different alveolar cells, inhaled particles and micro-organisms modulating pulmonary host defence systems . SP-A, the most abundant surfactant protein, binds to alveolar macrophages via a specific surface receptor with high affinity {128} . Such binding effects the release of reactive oxygen species from resident alveolar macrophages if SP-A is properly presented to the target cell . SP-A also stimulates chemotaxis of alveolar macrophages {142}, and serves as an opsonin in the phagocytosis of herpes simplex virus {161} Candida tropicalis {138} and various bacteria {137} . In addition, SP-A enhances the uptake of particles by monocytes and culture-derived macrophages {140} and improves bacterial killing . SP-D, another hydrophobic surfactant-associated protein, might interact with alveolar macrophages as well, stimulating the release of oxygen radicals {148}, while for the hydrophilic surfactant proteins SP-B and SP-C no macrophage interactions have been described so far . SP-A and SP-D are members of the so-called 'collectins', pattern recognition molecules involved in first line defence . While some surfactant proteins appear to stimulate certain macrophage defence functions, surfactant phospholipids seem to inhibit those of lymphocytes . Suppressed lymphocyte functions include lymphoproliferation in response to mitogens and alloantigens, B cell immunoglobulin production and natural killer cell cytotoxicity . Concerning surfactant's phospholipid composition phosphatidylglycerol is more suppressive than phosphatidylcholine on a molar basis {38} . Bovine surfactant has an immunosuppressive effect on the development of hypersensitivity pneumonitis in a guinea pig model {150} . Despite these interesting observations, several important questions concerning the interactions of surfactant components with pulmonary host defence systems remain unanswered . Sufficient host defence in the lungs works through various humoral-cellular systems in conjunction with the specific anatomy of the airways and the gas exchange surface--how does the surfactant system fit into this network? Surfactant and alveolar cells are both altered during lung injury--is there a relationship between alveolar cells from RDS patients and the endogenous surfactant isolated from such patients? How does exogenous surfactant as used for substitution therapy modulate the defence system of the host? Some of those artificial surfactants have been shown to inhibit the endotoxin-alveolar macrophages, PMNs and monocytes including IL-1, IL-6 and TNF {139,152}.(ABSTRACT TRUNCATED AT 400 WORDS)

J Mol Graph, 1994 Sep, 12(3), 185 - 92, 195
Modeling cytochrome P450 14 alpha demethylase (Candida albicans) from P450cam; Boscott PE et al.; The tertiary structure of cytochrome P450 14 alpha demethylase--Candida albicans (P450 CA) is modeled on the basis of sequence alignment with two closely related proteins and the crystallographic structure of Pseudomonas putida P450cam . The secondary structure prediction system used combines the information from several algorithms and trains the data to offer an optimized prediction of the known P450cam . The trained algorithm was then used to predict the secondary structure of the other P450 sequences . The prediction of the surface coil regions was aided by an alignment between P450 CA and the homologous sequences P450 14 alpha demethylase--Saccharomyces cerevisiae (66 SD) and P450 14 alpha demethylase--Candida tropicalis (72 SD) . The prediction and alignment information was combined to establish an alignment between P450 CA and P450cam, and to assign full secondary structure to the target protein . This secondary structure was folded from the template of P450cam and the predicted structure was relaxed by molecular dynamics . Model checking highlighted minor adjustments in the alignment, correctly orienting hydrophobic and hydrophilic side chains . The model offers explanations for several known experimental results and suggests further investigations that may prove fruitful in understanding the structure and mechanisms of the P450 family (Porter, T.D . and Coon, M.J . Minireview cytochrome P450 . J . Biol . Chem . 1991, 266, 13469-13472 . Waterman, M.R . Cytochrome P450 cellular distribution and structural considerations . Current Opinion in Structural Biology 1992, 2, 384-387 . Aoyama, Y., Yoshida, Y., Sonohdo, Y . and Sato, Y . Structural analysis of the interaction between the side-chain of substrates and the active site of lanosterol 14 alpha demethylase (P450 14DM) of yeast . Biochim . Biophys . Acta 1992, 1122, 251-255.).

J Clin Microbiol, 1994 Sep, 32(9), 2099 - 102
Comparison of broth macrodilution, broth microdilution, and E test antifungal susceptibility tests for fluconazole; Sewell DL et al.; A comparison of the E test, the broth microdilution test, and the reference broth macrodilution susceptibility test of the National Committee for Clinical Laboratory Standards for fluconazole susceptibility testing was performed with 238 clinical isolates of Candida species and Torulopsis (Candida) glabrata . An 80% inhibition endpoint MIC was determined by the reference broth macrodilution method after 48 h of incubation . The MICs obtained by the two study methods were read after 24 and 48 h of incubation . Overall, excellent agreement within 2 doubling dilutions was obtained between the broth microdilution and the broth macrodilution methods for the combined results for all species at both 24 h (93%) and 48 h (94%) . The correlation of 24-h MIC endpoints between the E test and the broth macrodilution methods was 37% for T . glabrata, 56% for Candida tropicalis, 93% for Candida albicans, and 90% for other Candida species . The percent agreement at 48 h ranged from 34% for T . glabrata to 97% for Candida species other than C . albicans and C . tropicalis . These initial results support the further evaluation of the E test as an alternative method for fluconazole susceptibility testing of Candida species.

Mycoses, 1994 Sep-Oct, 37(9-10), 343 - 7
Growth inhibition of pathogenic yeasts by Pseudomonas aeruginosa in vitro: clinical implications in blood cultures; Grillot R et al.; The interaction between yeasts and bacteria may have clinical implications in polymicrobial septicaemia . The in vitro effect of Pseudomonas aeruginosa on five pathogenic yeast species, Candida albicans, Candida tropicalis, Candida parapsilosis, Candida glabrata and Cryptococcus neoformans, was investigated . Yeast inhibition assays were performed in an aerobic blood culture medium, inoculated with yeast cells (inoculum 1-10 CFU ml-1) and bacterial cells (inoculum 10-10(7) CFU ml-1) . Interactions between P . aeruginosa and yeasts were determined after incubation by enumeration of pure and mixed cultures . Growth of all isolates tested was completely or partially inhibited by P . aeruginosa in blood culture medium, the phenomenon depending on the yeast genus and bacterial inoculum . Suppression of fungal growth was also observed in bacterial culture filtrate . This in vitro antifungal activity may preclude yeast recovery from blood cultures in mixed infections.

Biochemistry, 1994 Aug 16, 33(32), 9791 - 9
Extracellular aspartic proteinases from Candida albicans, Candida tropicalis, and Candida parapsilosis yeasts differ substantially in their specificities; Fusek M et al.; Extracellular aspartic proteinases have been implicated for some time as virulence factors associated with Candida opportunistic fungal infections . Our present knowledge of the enzymatic properties of these proteinases is rather limited . Information on their substrate specificity is important for understanding their roles in invasive Candida infections . We have isolated aspartic proteinases from each of the three Candida yeasts, Candida albicans, Candida tropicalis, and Candida parapsilosis, and investigated the specificities of these proteinases using a library of synthetic substrates and testing inhibition by pepstatin A . The specificities of these aspartic proteinases are different from those of major human proteinases, including gastric pepsins, renal renin, and cathepsin D . For the peptide substrate, Lys-Pro-Ala-Leu-Phe*Phe(p-NO2)-Arg-Leu, the values of kcat/Km were 2.95 x 10(6) M-1 s-1 for cleavage by Candida albicans proteinase, 1.60 x 10(6) M-1 s-1 for cleavage by Candida tropicalis proteinase, and 0.59 x 10(6) M-1 s-1 for Candida parapsilosis proteinase . Substantial differences in specificity among the Candida yeast proteinases were identified . For example, Candida tropicalis shows large changes in the kcat/Km value depending on the acidobasic character of the residue occupying the P2 position (1.6 x 10(6) M-1 s-1 for Leu, 0.47 x 10(6) M-1 s-1 for Lys, and 0.05 x 10(6) M-1 s-1 for Asp at P2, respectively) . Candida parapsilosis by comparison is tolerant of these substitutions at P2 and is highly restrictive at position P4.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemistry, 1994 Aug 16, 33(32), 9496 - 503
Specific adherence of Candida tropicalis to lysophospholipids; Prakobphol A et al.; Candida species are usually commensal organisms, but they become invasive when the host is immunocompromised . Mechanisms by which these organisms adhere to, colonize, and then invade host tissues are poorly understood . To detect potential host receptors, members of a lipid library were chromatographically separated and then overlaid with Candida tropicalis; components to which the organisms bound were visualized by autoradiography . In initial experiments no interactions with either glycolipids or intact phospholipids were detected . However, lysophospholipids supported adherence of C . tropicalis but not Saccharomyces cerevisiae . These results were confirmed by a second assay; C . tropicalis adhered to certain lysophospholipids, but not intact phospholipids, that were immobilized on microtiter plates . Using {14C}-1-palmitoyl-sn-glycero-3-phosphocholine, we showed that C . tropicalis adherence is accompanied by rapid conversion of the labeled lipid to a number of compounds . Thus, the interaction of C . tropicalis with lysophospholipids results in significant changes in both the organism and the lysophospholipid to which it binds . We hypothesize that this interaction could be an important component of the infection process.

Cancer, 1994 Aug 15, 74(4), 1360 - 6
A radiologic syndrome after high dose chemotherapy and autologous bone marrow transplantation, with clinical and pathologic features of systemic candidiasis; Mudad R et al.; BACKGROUND . The use of high dose chemotherapy in the treatment of solid tumors is associated with prolonged neutropenia and, consequently, in some patients, systemic candidiasis . The authors describe their experience with a clinicoradiologic syndrome developing after high dose chemotherapy was administered to patients with breast cancer . METHODS . The authors evaluated the clinical and radiologic records of 12 patients in whom hepatic, splenic, or renal candidiasis developed . RESULTS . Three patients had positive blood cultures for candida tropicalis . One of these patients and two others had fungal organisms identified with special stains of an organ aspirate . Most patients were asymptomatic, and most of them were treated successfully with antifungal agents, although untreated patients also recovered . There were no fatalities due to the candidiasis . CONCLUSIONS . A radiographic syndrome resembling hepatic, splenic, or renal candidiasis is described, which occurred after high dose chemotherapy was administered and autologous bone marrow transplantation was performed on patients with breast cancer . This syndrome has a favorable prognosis . Conclusions as to the more indolent nature of this syndrome cannot be made; however, this topic warrants further investigation.

Yeast, 1994 Aug, 10(8), 1065 - 74
Predominant localization of non-specific lipid-transfer protein of the yeast Candida tropicalis in the matrix of peroxisomes; Tan H et al.; PXP-18 is a 14-kDa major peroxisomal protein of the yeast Candida tropicalis and a homologue of the non-specific lipid-transfer protein (nsLTP) of mammals . Mammalian nsLTP is thought to facilitate the contact of membranes, to stimulate lipid-transfer between them . If PXP-18 functions like nsLTP, it must be present on organelle membranes . Immunoelectron microscopy of C . tropicalis cells indicated that gold particles, which visualized PXP-18, localized exclusively in the matrix of peroxisomes . Subcellular fractionation followed by Western blotting revealed the association of PXP-18 with peroxisomes in C . tropicalis cells . An enzyme-linked immunosorbent assay revealed that almost all the PXP-18 associated with peroxisomes was detectable after the solubilization of the organelle but not before, implying the predominance of PXP-18 inside peroxisomes . This differential assay was applied to the intracellular import of the intact and truncated PXP-18s expressed in Saccharomyces cerevisiae cells . Most of the intact PXP-18 was shown to be imported into the matrix of host-cell peroxisomes, whereas the truncated PXP-18, which lacked the C-terminal tripeptide Pro-Lys-Leu, no longer targeted peroxisomes . These results are consistent with the view that PXP-18 is the matrix protein of peroxisomes and must function in a system other than that of lipid transfer.

J Clin Microbiol, 1994 Aug, 32(8), 1902 - 7
Rapid detection and identification of Candida albicans and Torulopsis (Candida) glabrata in clinical specimens by species-specific nested PCR amplification of a cytochrome P-450 lanosterol-alpha-demethylase (L1A1) gene fragment; Burgener-Kairuz P et al.; PCR of a Candida albicans cytochrome P-450 lanosterol-alpha-demethylase (P450-L1A1) gene segment is a rapid and sensitive method of detection in clinical specimens . This enzyme is a target for azole antifungal action . In order to directly detect and identify the clinically most important species of Candida, we cloned and sequenced 1.3-kbp fragments of the cytochrome P450-L1A1 genes from Torulopsis (Candida) glabrata and from Candida krusei . These segments were compared with the published sequences from C . albicans and Candida tropicalis . Amplimers for gene sequences highly conserved throughout the fungal kingdom were first used; positive PCR results were obtained for C . albicans, T . glabrata, C . krusei, Candida parapsilosis, C . tropicalis, Cryptococcus neoformans, and Trichosporon beigelii DNA extracts . Primers were then selected for a highly variable region of the gene, allowing the species-specific detection from purified DNA of C . albicans, T . glabrata, C . krusei, and C . tropicalis . The assay sensitivity as tested for C . albicans in seeded clinical specimens such as blood, peritoneal fluid, or urine was 10 to 20 cells per 0.1 ml . Compared with results obtained by culture, the sensitivity, specificity, and efficiency of the species-specific nested PCR tested with 80 clinical specimens were 71, 95, and 83% for C . albicans and 100, 97, and 98% for T . glabrata, respectively.

J Chemother, 1994 Aug, 6(4), 226 - 9
Fluconazole, itraconazole and ketoconazole in vitro activity against Candida spp; Arevalo MP et al.; The in vitro activity of fluconazole, itraconazole and ketoconazole against 625 Candida yeast strains from patients treated at the University Hospital of the Canaries, by means of a micromethod of dilution in broth enriched with Yeast Nitrogen Base (YNB), and buffered to pH7, has been assessed . Species distribution was as follows: Candida albicans (388), Candida tropicalis (84), Candida glabrata (84), Candida parapsilosis (69) . Our results show 10.0% and 8.8% of C . albicans resistant to itraconazole and fluconazole, respectively, and 1.8% resistant to ketoconazole; 39.5% of C . tropicalis were resistant to itraconazole, 34.5% to fluconazole and 2.4% to ketaconazole . 19.1% of C . glabrata were resistant to fluconazole and 13.1% to itraconazole; 4.4% of C . parapsilosis were resistant to fluconazole and 1.5% to itraconazole . In general C . tropicalis was the most resistant strain and C . parapsilosis the most sensitive . The greatest percentages of resistance in vitro were seen with the two triazols.

Oral Microbiol Immunol, 1994 Aug, 9(4), 229 - 35
The in vitro proteolytic and saccharolytic activity of Candida species cultured in human saliva; Samaranayake YH et al.; The proteolytic and saccharolytic activity of 4 Candida species was investigated in batch cultures of pooled, human mixed saliva supplemented with glucose . All the Candida species investigated (Candida albicans, Candida tropicalis, Candida glabrata and Candida krusei) demonstrated a marked growth in saliva with a concomitant reduction in pH from about 7.5 to 3.3, within 72 h . Isotachophoretic analysis of the culture supernatant revealed the presence of a variety of acid anions of which pyruvate and acetate were the most abundant . Proteolysis of salivary components, evaluated by a biochemical assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was exhibited by all 4 Candida species, although there was inter-species variation . Despite the similarity in growth rates, C . tropicalis and C . krusei demonstrated greater proteolytic activity than C . albicans and C . glabrata . Neither candidal growth nor proteolysis was observed in glucose-free control saliva samples . In contrast, the degree of saccharolytic and proteolytic activity of a single isolate of C . albicans in glucose-supplemented parotid saliva appeared to be relatively weak compared with mixed saliva . As the oral cavity provides ideal low pH niches periodically supplemented with dietary carbohydrates, the acidic proteinases of Candida species may play a role in the pathogenesis of oral candidiasis.

Antimicrob Agents Chemother, 1994 Jul, 38(7), 1523 - 9
Synergism between the antifungal agents amphotericin B and alkyl glycerol ethers; Haynes MP et al.; The alkyl glycerol ether rac-1-O-dodecylglycerol inhibited the growth of members of two genera of yeasts, Candida and Cryptococcus, and was strongly synergistic with amphotericin B . At one-half its MIC, dodecylglycerol decreased the MIC of amphotericin B by as much as 80-fold . This high degree of synergism between dodecylglycerol and amphotericin B was demonstrated against a number of species of yeasts including Candida albicans, Candida tropicalis, Candida parapsilosis, Cryptococcus neoformans, Cryptococcus albidus, and Cryptococcus laurentii . All fractional inhibitory concentrations (for all strains and species) were calculated to be less than 1, and most were less than 0.6, again demonstrating strong synergism . Other alkyl glycerol ethers with alkyl chain lengths ranging from 8 to 18 carbon atoms were also found to be synergistic with amphotericin B against C . neoformans and C . albicans . Electron microscopy experiments showed that C . neoformans grown in the presence of dodecylglycerol had severely abnormal, deformed capsules . Although the mechanism of action of dodecylglycerol is not known, dodecylglycerol was not simply acting as a detergent . The natural detergent sodium deoxycholate could not substitute for dodecylglycerol . At comparable and higher concentrations, sodium deoxycholate had no fungicidal effect on its own, nor did it potentiate the activity of amphotericin B . Dodecylglycerol did not interact synergistically with the water-soluble antifungal agent fluconazole . The lipid-soluble hydrophobic properties of amphotericin B appear to be important for this synergistic effect, in that alkyl glycerol ethers could promote synergism with amphotericin B by potentially increasing the interaction between membrane-bound ergosterol and amphotericin B.

J Clin Microbiol, 1994 Jul, 32(7), 1650 - 3
Selection of candidate quality control isolates and tentative quality control ranges for in vitro susceptibility testing of yeast isolates by National Committee for Clinical Laboratory Standards proposed standard methods; Pfaller MA et al.; The National Committee for Clinical Laboratory Standards has developed a proposed standard method for in vitro antifungal susceptibility testing of yeast isolates (National Committee for Clinical Laboratory Standards, document M27-P, 1992) . In order for antifungal testing by the M27-P method to be accepted, reliable quality control (QC) performance criteria must be developed . In the present study, five laboratories tested 10 candidate QC strains 20 times each against three antifungal agents: amphotericin B, fluconazole, and 5-fluorocytosine . All sites conformed to the M27-P standards and used a common lot of tube dilution reagents and RPMI 1640 broth medium . Overall, 98% of MIC results with amphotericin B, 95% with fluconazole, and 92% with 5-fluorocytosine fell within the desired 3-log2 dilution range (mode +/- 1 log2 dilution) . Excellent performance with all three antifungal agents was observed for six strains: Candida albicans ATCC 90028, Candida parapsilosis ATCC 90018, C . parapsilosis ATCC 22019, Candida krusei ATCC 6258, Candida tropicalis ATCC 750, and Saccharomyces cerevisiae ATCC 9763 . With these strains, 3-log2 dilution ranges encompassing 94 to 100% of MICs for all three drugs were established . Additional studies with multiple lots of RPMI 1640 test medium will be required to establish definitive QC ranges.

Eur J Clin Microbiol Infect Dis, 1994 Jul, 13(7), 590 - 5
An experimental model for study of Candida survival and transmission in human volunteers; Rangel-Frausto MS et al.; In order to determine the potential for cross-transmission of Candida spp . between health-care workers and patients, the survival of clinical isolates of five species of Candida on the palms of human volunteers was tested . One hundred microliters of a McFarland 1.0 density suspension (5 x 10(5) cfu) from an overnight culture of Candida albicans, Candida krusei, Candida parapsilosis, Candida tropicalis and Candida glabrata was used as inoculum . The degree of hydrophobicity of the different Candida species was also tested and did not influence the survival . The half-lives were brief, being 9.5, 12.4, 7.4, 12.8, 9.6 min for Candida albicans, Candida krusei, Candida glabrata, Candida parapsilosis, and Candida tropicalis, respectively, but at 45 min 2.6 x 10(3) to 3 x 10(4) organisms remained on the hands . Survival of Candida albicans for as long as 24 h on inanimate surfaces was observed . Transmission from one hand to a second hand occurred in 69% of the experiments and from the first to a third hand in 38% . Transmission to and from inanimate surfaces was successful in most of the experiments (90%) . This experimental model aids in the biological study of Candida spp . and suggests some of the potential mechanisms of transmission.

Mycoses, 1994 Jul-Aug, 37(7-8), 285 - 9
In vitro susceptibility of 545 isolates of Candida spp . to four antifungal agents; Arias A et al.; The in vitro susceptibility to amphotericin B, fluconazole, itraconazole and ketoconazole of 545 Candida strains from patients treated at the University Hospital of the Canaries was determined by means of a microdilution test . The distribution of the species was as follows: Candida albicans (342), Candida tropicalis (70), Candida glabrata (68), Candida parapsilosis (65) . Of Candida albicans isolates, 8.5% and 7.6% showed resistance to itraconazole and fluconazole respectively . Of C . tropicalis isolates 34.3%, 27.1% and 2.9% were resistant to itraconazole, fluconazole and ketaconazole respectively . For C . glabrata, 10.3% and 4.4% of the isolates under study demonstrated resistance to fluconazole and itraconazole respectively . Only 4.6% and 1.5% of C . parapsilosis isolates demonstrated resistance to fluconazole and itraconazole respectively . C . tropicalis was the most resistant strain and C . parapsilosis the most sensitive . The greatest percentages of resistance in vitro were seen with the triazoles.

APMIS, 1994 Jun, 102(6), 451 - 6
Mycotic and algal bovine mastitis in Denmark; Aalbaek B et al.; A one-year examination of mammary secretions (n = 2,896) from Danish cattle with clinical or subclinical mastitis revealed 45 strains of fungi and algae . The strains originated from 44 mammary secretions of 42 cows in 40 herds . The following species of fungi were identified: Candida catenulata (n = 2), Candida kefyr (n = 6), Candida krusei (n = 17), Candida rugosa (n = 6), Candida tropicalis (n = 3), Candida valida (n = 1), Geotrichum capitatum (n = 5) . The algal species Prototheca zopfii was demonstrated in five samples.

Ophthalmology, 1994 Jun, 101(6), 1005 - 13
The changing spectrum of fungal keratitis in south Florida; Rosa RH Jr et al.; PURPOSE: To review the clinical experience with fungal keratitis in south Florida over a 10-year period . METHODS: One hundred twenty-five cases of fungal keratitis were identified in the microbiology laboratory records between January 1982 and January 1992 . The medical record of each patient was reviewed . RESULTS: The most commonly associated risk factor was trauma (44%) . Fungal keratitis developed in five patients using extended wear contact lenses and one patient wearing a therapeutic bandage contact lens . Clinical features included irregular, feathery margins (62%), a dry, rough texture (47%), and satellite lesions (41%) . An initial positive culture was obtained in 90% of patients, with a majority of cultures becoming positive within 48 hours . The Fusarium sp accounted for 62% of the isolates, with Fusarium oxysporum being the most commonly isolated organism . New fungal isolates include Candida parapsilosis, Aspergillus terreus, Candida tropicalis, and Trichosporon beigellii . Natamycin 5% suspension was the initial antifungal agent used for 91% of the patients, with an average duration of treatment of 38 days . Twenty-five patients were treated with oral ketoconazole for a median duration of 2 weeks, in addition to topical antifungal therapy . Thirty-four patients (27%) required a penetrating keratoplasty . Six patients had recurrence of fungal keratitis after penetrating keratoplasty . CONCLUSIONS: Trauma, including contact lens wear, is the most commonly associated risk factor . The fungal organisms can be readily identified in culture . F . oxysporum is the most common organism, with new isolates identified . The mainstay of therapy is topical natamycin with the increasing use of imidazoles.

J Clin Microbiol, 1994 May, 32(5), 1184 - 7
Comparison of updated Vitek Yeast Biochemical Card and API 20C yeast identification systems; Fenn JP et al.; The updated Vitek Yeast Biochemical Card (YBC) was compared with the API 20C by using 409 germ tube-negative yeasts and Geotrichum spp . that were either clinical or proficiency sample isolates . The API 20C was the reference standard . The 409 isolates represented nine genera and 21 species . Morphology agars were inoculated and interpreted for each isolate . The API 20C identified 406 isolates (99.3%), while the Vitek YBC identified 367 (89.7%) . Both systems identified the majority of yeasts after 24 h of incubation--73.4% were identified by the API 20C and 77.4% were identified by the Vitek YBC . The Vitek 24-h reading had some incorrect identifications . These included 14 isolates of Candida tropicalis that were identified as Candida parapsilosis (91 to 97% reliability) and 3 isolates of Candida krusei that were called Blastoschizomyces capitatus (Geotrichum capitatum), Candida rugosa, and Candida zeylanoides . In total, the Vitek YBC misidentified 30 isolates, while the API 20C misidentified 3 isolates . In addition, results for 14 isolates with the Vitek YBC were listed under the category "no identification." Morphology agars were required for identification with 89 isolates (21.9%) when the API 20C was used and with 50 isolates (12.6%) when the Vitek YBC was used . Apart from the price of the Vitek instrument, the API 20C costs $1.28 more per test than the Vitek YBC . Overall, the updated Vitek YBC compares favorably with the API 20C in the identification of common yeasts such as Torulopsis glabrata, C . parapsilosis, and Cryptococcus neoformans . However, problems were encountered with the Vitek system in the identification of C . tropicalis, C . krusei, Trichosporon spp., and some Cryptococcus spp . The routine use of morphology agars with either method is recommended.

J Infect, 1994 May, 28(3), 305 - 10
Inhibition of fungal growth by Pseudomonas aeruginosa and Pseudomonas cepacia isolated from patients with cystic fibrosis; Kerr J; This study was undertaken because of the infrequency of infections due to Candida species in patients with cystic fibrosis despite their extensive treatment with broad-spectrum antibiotics . In vitro susceptibility studies revealed significant inhibition of 11 strains of fungi known to infect human beings by 10 strains of Pseudomonas aeruginosa and nine strains of Pseudomonas cepacia isolated from the sputum of patients with cystic fibrosis . The fungi were Candida krusei, Candida keyfr, Candida guillermondii, Candida tropicalis, Candida lusitaniae, Candida parapsilosis, Candida pseudotropicalis, Candida albicans, Torulopsis glabrata, Saccharomyces cerevisiae and Aspergillus fumigatus . Inhibition of fungal growth by Escherichia coli (NCTC 10418), Staphylococcus aureus (NCTC 6571) and Haemophilus influenzae (NCTC 11931) could not be demonstrated . The continued presence in the sputum of patients with cystic fibrosis of strains of P . aeruginosa and P . cepacia, which produce antifungal substances, may inhibit growth of Candida species and so prevent overt Candida infections . A . fumigatus would seem to be the most important fungus causing disease in patients with cystic fibrosis . It is therefore interesting to note that this was the most resistant of all the fungi tested for inhibition by P . aeruginosa and P . cepacia.

Proc Natl Acad Sci U S A, 1994 Apr 12, 91(8), 3453 - 7
A conserved sequence motif within the exceptionally diverse telomeric sequences of budding yeasts; McEachern MJ et al.; Telomeric DNA sequences have generally been found to be remarkably conserved in evolution, typically consisting of repeated, very short sequence units containing clusters of G residues . Recently however the telomeric DNA of the asexual yeast Candida albicans was shown to consist of much longer repeat units . Here we report the identification of seven additional telomeric sequences from sexual and asexual budding yeast species . The telomeric repeat units from this group of relatively closely related species show more phylogenetic diversity in length (8-25 bp), sequence, and composition than has been seen previously throughout a wide phylogenetic range of other eukaryotes . We also show that certain strains of the asexual diploid species Candida tropicalis have two forms of telomeric repeats, which appear to differ by a single base pair . Despite their great diversity, the telomeric repeat units of C . albicans, Saccharomyces cerevisiae, and all of the species we have examined in this report share a conserved approximately 6-bp motif of T and G residues resembling more typical telomeric sequences.

Am J Clin Pathol, 1994 Apr, 101(4), 438 - 42
Clinical comparison of the Baxter MicroScan Yeast Identification Panel and the Vitek Yeast Biochemical Card; Riddle DL et al.; To determine the reliability of the Baxter MicroScan Yeast Identification Panel, processed by the Walkaway-96, and the Vitek Yeast Biochemical Card, 150 clinical yeast isolates (30 Candida albicans, 67 Candida species, not albicans, 26 Torulopsis glabrata, 13 Cryptococcus neoformans, 4 Saccharomyces cerevisiae, 6 Trichosporon beigelii, 3 Rhodotorula species, and 1 Geotrichum species) were tested on both systems . Results were compared with those obtained by the API 20C and the appearance of yeast cells on cornmeal Tween-80 agar . After inoculation of each system, results were available in 4 hours with MicroScan panels, 24-48 hours with Vitek cards, and 72 hours with the API 20C strips . On initial testing, 101 (67%) and 128 (85%) isolates, respectively, were correctly identified by MicroScan and Vitek . After repeat testing, the number of correctly identified isolates increased to 123 (82%) by MicroScan and to 142 (95%) by Vitek . Yeasts most commonly misidentified were Candida tropicalis, T glabrata, and Candida parapsilosis by MicroScan and C tropicalis and T glabrata by Vitek.

Infect Immun, 1994 Apr, 62(4), 1489 - 93
Phagocytosis of medically important yeasts by polymorphonuclear leukocytes; Lyman CA et al.; Phagocytosis is a critical function of polymorphonuclear leukocytes in the control of mycotic infections . By using a modified fluorescence quenching assay to distinguish between attached and ingested organisms, we determined the percent phagocytosis of several medically important yeasts . The percentages of phagocytosis of serum-opsonized Candida albicans, Candida tropicalis, Candida parapsilosis, and Torulopsis glabrata were all comparable at 37 degrees C . By comparison, there was significantly less phagocytosis of Cryptococcus neoformans and Trichosporon beigelii isolates (P < 0.001) . Thus, phagocytosis of C . albicans by polymorphonuclear leukocytes is comparable to that of species other than C . albicans but is significantly greater than that of the basidiomycetous yeasts T . beigelii and C . neoformans.

FEBS Lett, 1994 Mar 28, 342(1), 19 - 22
Expression of alpha-1,3 linkage-containing oligomannosyl residues in a cell-wall mannan of Candida tropicalis grown in yeast extract-Sabouraud liquid medium under acidic conditions; Kobayashi H et al.; We investigated the cell-wall mannan obtained from Candida tropicalis IFO 1647 strain cells grown in yeast extract-Sabouraud medium at pH 3.0 by two-dimensional homonuclear Hartmann-Hahn spectroscopy . The results indicate that the phosphate group and the side chains containing a beta-1,2-linked mannopyranose unit decreased compared to those of mannan from cells grown under conventional conditions (pH 5.9) with concomitant expression of alpha-1,3 linkage-containing oligomannosyl side chains . The results of acetolysis of these mannans indicated that the presence of alpha-1,3-linked mannopyranose unit existed in side chains corresponding to pentaose and hexaose, Manp alpha 1-3 Manp alpha 1-2Manp alpha 1-2 Manp alpha 1-2Man, and Manp alpha 1-2Manp alpha 1-3Manp alpha 1-2Manp alpha 1-2Manp alpha 1-2Man, in the mannan from cells grown at pH 3.0.

Clin Infect Dis, 1994 Mar, 18(3), 440 - 2
An unusual cause of acute renal failure: bilateral ureteral obstruction due to Candida tropicalis fungus balls; Scerpella EG et al.; Fungus balls have rarely been implicated as a cause of urinary tract obstruction . Approximately 50 cases of fungus balls of the urinary tract have been reported previously; the majority of cases were characterized by unilateral ureteral involvement or bladder involvement, and Candida albicans has been the organism most frequently isolated . We report, to our knowledge, the first case of bilateral ureteral obstruction caused by Candida tropicalis fungus balls.

Mycoses, 1994 Mar-Apr, 37(3-4), 101 - 7
Pathogenicity of yeasts and algae isolated from bovine mastitis secretions in a murine model; Jensen HE et al.; The pathogenicity of strains of yeasts and algae isolated from bovine mastitis secretions was evaluated in immunosuppressed mice . Strains of Candida tropicalis (n = 3) were the most pathogenic, but all strains of Geotrichum capitatum (n = 5) and the colourless alga Prototheca zopfii (n = 5) were also lethal to mice at the highest dose of 1 x 10(7) CFU per mouse . Reisolation of the inoculated micro-organisms and the occurrence of histopathological lesions within organs of mice challenged with 1 x 10(3) to 1 x 10(7) CFU per animal revealed strains of C . krusei (n = 17), C . kefyr (n = 6) and C . rugosa (n = 6) to have a moderate pathogenicity, whereas strains of C . valida (n = 1) and C . catenulata (n = 2) were weakly pathogenic and non-pathogenic respectively.

J Clin Microbiol, 1994 Feb, 32(2), 525 - 7
Suppression of fungal growth exhibited by Pseudomonas aeruginosa; Kerr JR; Three surgery patients were monitored postoperatively, with particular reference to lung infection . In each case there was a clinical impression that Pseudomonas aeruginosa suppressed the growth of Candida albicans in patients with clinically significant lung infections from whom both of these organisms were isolated from serial sputum samples . Regrowth of C . albicans after P . aeruginosa eradication occurred in two patients, despite fluconazole therapy, to which both C . albicans isolates were susceptible . In all three patients, the strain of P . aeruginosa was found to inhibit the growth of the corresponding C . albicans strain in vitro . Further in vitro susceptibility studies revealed significant inhibition by 10 strains of P . aeruginosa of 11 strains of fungi known to infect humans; these were Candida krusei, Candida keyfr, Candida guillermondii, Candida tropicalis, Candida lusitaniae, Candida parapsilosis, Candida pseudotropicalis, Candida albicans, Torulopsis glabrata, Saccharomyces cerevisiae, and Aspergillus fumigatus.

Infect Immun, 1994 Feb, 62(2), 615 - 22
Structures of cell wall mannans of pathogenic Candida tropicalis IFO 0199 and IFO 1647 yeast strains; Kobayashi H et al.; We conducted a structural analysis of the cell wall mannans isolated from two Candida tropicalis strains, IFO 0199 and IFO 1647, exhibiting strong agglutinabilities against anti-Candida factor sera 5 and 6 . The products released from these mannans by acid treatment were identified as the oligosaccharides, from biose to pentaose, consisting solely of beta-1,2-linked mannopyranose units corresponding to common epitopes of Candida albicans serotypes A and B (factor 5) . Mild acetolysis of acid- and alkali-treated mannans produced large amounts of hexaose and heptaose, Man rho beta 1-2Man rho beta 1-2Man rho alpha 1-2Man rho alpha 1-2Man rho alpha 1-2Man and Man rho beta 1-2Man rho beta 1-2Man rho beta 1-2Man rho alpha 1-2 Man rho alpha 1-2Man, corresponding to the C . albicans serotype A-specific epitopes (factor 6) . However, the homologous pentaose, Man rho beta 1-2Man rho alpha 1-2 Man, was not generated by this procedure . The oligosaccharides (biose to hexaose) obtained from the mannans by conventional acetolysis were composed exclusively of alpha-1,2-linked mannopyranose units . Therefore, the mannans of C . tropicalis IFO 0199 and IFO 1647 do not have the alpha-1,3-linked mannopyranose units previously observed in the mannans of C . albicans and Candida stellatoidea . The results of this study and previous findings indicate that the similarity of the antigenicities of three Candida species, C . albicans serotype A, C . stellatoidea type II, and C . tropicalis, reside in the beta-1,2 and alpha-1,2 linkages containing oligomannosyl side chain (factor 6) in the cell wall mannan.

Ned Tijdschr Geneeskd, 1994 Jan 1, 138(1), 35 - 8
{Hepatosplenic candidiasis in patients treated for hemato-oncological disorders}; Zweegman S et al.; Hepatosplenic candidiasis is increasingly observed in patients with a haematological malignancy who have received chemotherapy . A case history is described of a male aged 45 who developed symptoms of hepatosplenic candidiasis caused by Candida tropicalis after treatment for acute myeloid leukaemia . The disease is characterized by persistent fever after recovery of the leukopenia induced by the chemotherapy . Echographic and computer-tomographic examination may reveal abscess patterns specific of Candida in the liver . Treatment consists of amphotericin B intravenously or fluconazole orally . Protracted treatment is frequently required.

J Clin Microbiol, 1994 Jan, 32(1), 92 - 7
An automated enzymatic method for measurement of D-arabinitol, a metabolite of pathogenic Candida species; Switchenko AC et al.; An automated enzymatic method was developed for the measurement of D-arabinitol in human serum . The assay is based on a novel, highly specific D-arabinitol dehydrogenase from Candida tropicalis . This enzyme catalyzes the oxidation of D-arabinitol to D-ribulose and the concomitant reduction of NAD+ to NADH . The NADH produced is used in a second reaction to reduce p-iodonitrotetrazolium violet (INT) to INT-formazan, which is measured spectrophotometrically . The entire reaction sequence can be performed automatically on a COBAS MIRA-S clinical chemistry analyzer (Roche Diagnostic Systems, Inc., Montclair, N.J.) . Replicate analyses of human sera supplemented with D-arabinitol over a concentration range of 0 to 40 microM demonstrated that the pentitol could be measured with an accuracy of +/- 7% and a precision (standard deviation) of +/- 0.4 microM . Serum D-arabinitol measurements correlated with those determined by gas chromatography (r = 0.94) . The enzymatic method is unaffected by L-arabinitol, D-mannitol, or other polyols commonly found in human serum . Any of 17 therapeutic drugs potentially present in serum did not significantly influence assay performance . Data illustrating the application of the assay in patients for possible diagnosis of invasive candidiasis and the monitoring of therapeutic intervention are presented . The automated assay described here was developed to facilitate the investigation of D-arabinitol as a serum marker for invasive Candida infections.

J Med Vet Mycol, 1994, 32(2), 115 - 22
Rapid presumptive identification of medically relevant yeasts to the species level by polymerase chain reaction and restriction enzyme analysis; Maiwald M et al.; A method for the rapid presumptive differentiation of a panel of 12 clinically relevant yeasts to the species level was developed on the basis of evaluation by the polymerase chain reaction (PCR) of the gene coding for the small ribosomal subunit 18S-rRNA . The method involved restriction enzyme analysis of PCR products obtained with primers common to all fungi . Using six restriction enzymes, AluI, BanI, BbsI, DraII, Eco147I and NheI, characteristic PCR-restriction enzyme patterns were obtained for Candida albicans, Candida tropicalis, Candida krusei, Candida kefyr, Candida lusitaniae, Candida guilliermondii, Candida glabrata and Saccharomyces cerevisiae, as well as for the pairs Candida parapsilosis/Candida viswanathii and Trichosporon beigelii/Cryptococcus neoformans . The procedure does not involve hybridization steps or the use of radioactivity and can be completed within one working day.

Gynecol Obstet Invest, 1994, 37(3), 164 - 7
In vitro infection of human fetal membranes with Candida species; Gurgan T et al.; To assess whether Candida species can penetrate intact fetal membranes under in vitro conditions, Candida albicans, Candida tropicalis, Candida guilliermondii, Candida pseudotropicalis and Candida glabrata were inoculated onto the surface of the maternal side of the membranes obtained from 4 pregnant women undergoing repeat cesarean section . After incubation under culture conditions, membranes were evaluated by histological examination . C . albicans inoculated onto the maternal side penetrated and passed to the fetal side and caused some degeneration of the structure of the membrane epithelium . The other four Candida species grew heavily on the maternal surface but did not penetrate and invade the membranes . This effect of C . albicans on fetal membranes may explain the potential mechanism in the development of Candida infections of the amniotic fluid, fetal membranes and possibly the fetus.

Med Dosw Mikrobiol, 1994, 46(3), 225 - 31
{Utilization of enzymatic activity in strains of Candida isolated from the vagina for typing and evaluation of pathogenicity}; Krzeminska-Jaskowiak E et al.; The study involved 255 strains of Candida isolated from vaginal secretions (194 with symptomatic mycosis and 61 from asymptomatic cases) . Most frequently isolated species were Candida albicans, Candida krusei and Candida tropicalis . Determination of lipolytic properties at 30 degrees C and 37 degrees C and proteolytic abilities against albumin and casein permitted for differentiation of strains belonging to Candida albicans and remaining strains of other Candida species . This resulted also in determination of biotypes . Strains of Candida albicans belonged mainly to biotype I, while strains of Candida krusei and Candida tropicalis to biotype IX . Worth attention are results regarding symptomic candidiasis of vagina and enzymatic properties of Candida albicans . Among 145 strains of Candida albicans isolated from cases of symptomatic vaginal mycosis 67 (46.2%) exhibited both lipolytic (at 30 degrees C and 37 degrees C) and proteolytic activities against albumin and casein . At the same time, in asymptomatic mycosis only 14 (33.3%) strains of Candida albicans exhibited same properties.

J Med Vet Mycol, 1994, 32(4), 257 - 65
Acid proteinase secreted by Candida tropicalis: virulence in mice of a proteinase negative mutant; Togni G et al.; The relationship between the ability to secrete a specific acid proteinase (ACP) by Candida tropicalis in the presence of bovine serum albumin as a nitrogen source and virulence for mice was studied using two stable proteinase-positive and proteinase-negative strains (DSY68 and DSY65), which were constructed from the wild-type pathogenic yeast C . tropicalis ATCC 750 . The inactivation of the gene encoding the secreted acid proteinase was produced by targeted gene disruption . Mortality rate was slightly lower in groups of mice infected with the proteinase-negative mutant . All other parameters analysed were similar for two strains of yeast . Our results therefore conclude that the ACP secreted by C . tropicalis did not contribute significantly to fungal virulence in systemic infections.

Ann Biol Clin (Paris), 1994, 52(6), 443 - 6
Evaluation of the ATB 32C, automicrobic system and API 20C using clinical yeast isolates; Gutierrez J et al.; The ATB 32C (bioMerieux, Spain), AMS-YBC (Vitek System, bioMerieux, Spain) and API 20C (bioMerieux, Spain) systems were evaluated for their reliability in identifying 100 clinical yeast isolates . The ATB 32C, AMS-YBC and API 20C systems correctly identified 97%, 98% and 100% of the isolates respectively . There were no significant differences in incubation periods between ATB 32C and AMS-YBC systems . One isolate of Candida tropicalis was wrongly identified by the ATB 32C and the AMS-YBC systems . The Saccharomyces cerevisiae isolate was wrongly identified by the ATB 32C system while the AMS-YBC failed to identify it and a third isolate of Candida krusei was wrongly identified by the ATB 32C system . The overall accuracy and rapidity of the ATB 32C and AMS-YBC systems were sufficient to permit recommendation of either of these systems for routine use in the clinical microbiology laboratory, although the first system enjoys the advantages of having a wider data-base and the possibility of manual reading.

Appl Microbiol Biotechnol, 1994 Jan, 40(5), 682 - 6
Characterization of the catalase of the n-alkane-utilizing yeast Candida tropicalis functionally expressed in Saccharomyces cerevisiae; Kinoshita H et al.; Candida tropicalis catalase (CTC) genomic DNA was recombined on a plasmid with the galactose-inducible GAL7 promoter and expressed highly as a heme protein in Saccharomyces cerevisiae as host . The percentage of recombinant CTC (rCTC) in total extractable protein amounted to at least 25% . The rCTC was purified and characterized in terms of subunit mass, behavior in native polyacrylamide gel electrophoresis, absorption spectrum, amino-terminal amino acid sequence, peptide map, specific activity, and Michaelis constant (Km) value for hydrogen peroxide . These properties were similar or identical to those of the purified enzyme from C . tropicalis (CTC) . From these results, this system appears suitable for high expression of functional catalase protein having heme.

J Clin Microbiol, 1993 Dec, 31(12), 3142 - 6
Use of a monoclonal antibody in a dot immunobinding assay for detection of a circulating mannoprotein of Candida spp . in neutropenic patients with invasive candidiasis; De Bernardis F et al.; A dot immunobinding assay for the detection of a circulating mannoprotein (MP) antigen of Candida species in the sera of neutropenic patients in a hematological setting is described . The technique is based on the use of a monoclonal antibody which recognizes an oligomannoside epitope shared by distinct MP of pathogenic Candida species . The sensitivity of the assay for antigen detection in serum was between 2 and 5 ng/ml, and MPs from Candida albicans, Candida tropicalis, Torulopsis glabrata, and Candida parapsilosis, but not Candida krusei, could be detected . A retrospective analysis of sera from patients with proven invasive candidiasis versus sera from controls (Candida-colonized and noncolonized subjects) revealed that the novel assay has sufficient sensitivity, specificity, and predictive values to be of potential diagnostic significance.

Biochem Biophys Res Commun, 1993 Nov 15, 196(3), 1323 - 9
Identification, purification, and characterization of a D-arabinitol-specific dehydrogenase from Candida tropicalis; Quong MW et al.; A novel D-arabinitol (DA) dehydrogenase was identified and purified more than 300-fold from Candida tropicalis . The enzyme is specific for DA and catalyzes the NAD(+)-dependent oxidation at carbon 4 to yield D-ribulose . Purification was accomplished by a combination of protamine sulfate and ammonium sulfate precipitation and dye ligand chromatography on a reactive yellow 86 column . The apparent Km of the enzyme for DA ({NAD+} = 2.2 mM) is 39.8 mM . The apparent Km for NAD+ ({DA} = 384 mM) is 0.12 mM . The pH-optimum for the enzymatic oxidation of DA is approximately 10 . Cofactor stereospecificity studies demonstrate that the enzyme catalyzes transfer of the 4(S) hydrogen of NADH with D-ribulose as substrate . The polyol substrate specificity of the present DA dehydrogenase makes the enzyme potentially useful for the development of a simple and specific method for the measurement of DA, a metabolite of pathogenic Candida spp . which has been described as a marker for disseminated candidiasis.

J Eukaryot Microbiol, 1993 Nov-Dec, 40(6), 733 - 41
Molecular genetic distinction of Pneumocystis carinii from rats and humans; Stringer JR et al.; Pneumocystis carinii from rats and from humans were compared with respect to electrophoretic karyotype, presence of DNA sequences known to be repeated in rat-derived P . carinii, overall DNA sequence homology, and the sequences at two genetic loci . The organisms from each host species were different in each respect . Neither of two repeated DNAs from rat-derived P . carinii was found in the genome of human-derived organisms, and total DNA from rat-derived P . carinii failed to hybridize to human-derived P . carinii DNA . The sequences of the alpha-tubulin genes from the two P . carinii were strikingly different and the base composition of the alpha-tubulin gene from rat-derived P . carinii was rich in adenine and thymine, while the base composition of this gene from human-derived P . carinii was rich in guanine and cytosine . The sequence from the 18S rRNA gene of human-derived P . carinii was twice as divergent from that of rat-derived P . carinii as the sequence from the corresponding region of Candida albicans was from that of Candida tropicalis . These data show that rats and humans can harbor distinct types of P . carinii that are sufficiently different to suggest that P . carinii from the two hosts could be different species.

Mol Gen Genet, 1993 Nov, 241(3-4), 422 - 30
Cloning of the isocitrate lyase gene (ICL1) from Yarrowia lipolytica and characterization of the deduced protein; Barth G et al.; The ICL1 gene encoding isocitrate lyase was cloned from the dimorphic fungus Yarrowia lipolytica by complementation of a mutation (acuA3) in the structural gene of isocitrate lyase of Escherichia coli . The open reading frame of ICL1 is 1668 bp long and contains no introns in contrast to currently sequenced genes from other filamentous fungi . The ICL1 gene encodes a deduced protein of 555 amino acids with a molecular weight of 62 kDa, which fits the observed size of the purified monomer of isocitrate lyase from Y . lipolytica . Comparison of the protein sequence with those of known pro- and eukaryotic isocitrate lyases revealed a high degree of homology among these enzymes . The isocitrate lyase of Y . lipolytica is more similar to those from Candida tropicalis and filamentous fungi than to Saccharomyces cerevisiae . This enzyme of Y . lipolytica has the putative glyoxysomal targeting signal S-K-L at the carboxy-terminus . It contains a partial repeat which is typical for eukaryotic isocitrate lyases but which is absent from the E . coli enzyme . Surprisingly, deletion of the ICL1 gene from the genome not only inhibits the utilization of acetate, ethanol, and fatty acids, but also reduces the growth rate on glucose.

Mycoses, 1993 Nov-Dec, 36(11-12), 417 - 20
Evaluation of the albicans IDR plate method for the rapid identification of Candida albicans; Lipperheide V et al.; Albicans IDR is a ready-to-use medium containing a hexosaminic substrate that allows rapid and specific identification of Candida albicans . In this study, we have evaluated its performance using 330 pure cultures of yeast strains previously identified by conventional mycological methods . The test correctly identified all the Candida albicans strains tested (186 isolates), but there were 28 false-positive reactions after 48 h of culture incubation . The false-positive isolates corresponded to the species Candida tropicalis (13 strains), Trichosporon beigelii (12 strains), Cryptococcus neoformans (one strain), Candida lusitaniae (one strain) and Candida membranaefaciens (one strain) . The sensitivity of the method was 100% and the specificity was 90.3% at 24 h and 86.6% at 48 h of incubation . Albicans IDR plate is a new, easy-to-perform and inexpensive method for C . albicans identification, and it could be used for direct isolation and identification of this species from clinical samples.

J Clin Invest, 1993 Oct, 92(4), 1840 - 9
Distinct mechanisms of epithelial adhesion for Candida albicans and Candida tropicalis . Identification of the participating ligands and development of inhibitory peptides; Bendel CM et al.; The yeast Candida albicans is the leading cause of disseminated fungal infection in neonates, immunocompromised hosts, diabetics, and postoperative patients; Candida tropicalis is the second most frequent isolate . Because the integrin analogue in C . albicans shares antigenic, structural, and functional homologies with the beta 2-integrin subunits alpha M and alpha X, we investigated the role of integrin analogues in epithelial adhesion of C . albicans and C . tropicalis . On flow cytometry with the monoclonal antibody (mAb) OKM1, surface fluorescence was highest for C . albicans and significantly reduced for C . tropicalis (P < 0.001) . However, adhesion to the human epithelial cell line HeLa S3 did not differ for these two candidal species: specific adhesion was highest for C . albicans at 44.0 +/- 1.8%, and only slightly lower for C . tropicalis at 38.8 +/- 3.6% (P = NS) . The disparity between expression of the integrin analogue and epithelial adhesion suggested distinct mechanisms for this process in C . albicans versus C . tropicalis . Preincubation of C . albicans with anti-alpha M mAbs, with purified iC3b (the RGD ligand for the integrin analogue), or with 9-15-mer RGD peptides from iC3b all inhibited epithelial adhesion significantly (P < 0.001-0.04) . Purified fibronectin or fibronectin-RGD peptides failed to block C . albicans adhesion . In contrast, epithelial adhesion of C . tropicalis was significantly inhibited by purified fibronectin and its RGD peptides (P < or = 0.021), but not by iC3b nor the iC3b-RGD peptides . Both iC3b and fibronectin were identified on the surface of epithelial cells after growth in serum-free medium . A polyclonal antibody to C3 inhibited C . albicans adhesion while a control antibody to fibronectin was ineffective; the converse was true for C . tropicalis . These results indicate that the pathogenic yeasts C . albicans and C . tropicalis recognize distinct RGD ligands present at the surface of the epithelial cell and that these interactions can be differentially inhibited by defined RGD peptides containing appropriate flanking sequences.

Infect Control Hosp Epidemiol, 1993 Oct, 14(10), 587 - 90
Outbreak of Candida tropicalis fungemia in a neonatal intensive care unit; Finkelstein R et al.; OBJECTIVE: To describe an outbreak of Candida tropicalis fungemia in a neonatal intensive care unit (NICU), to evaluate the risk factors associated with this infection and the possible mode of nosocomial transmission . DESIGN: Descriptive and case-control study . PATIENTS AND METHODS: Surveillance cultures were taken from hospitalized patients, personnel, and inanimate objects in the NICU . Six patients with C tropicalis fungemia (cases) were compared with C tropicalis culture-negative patients matched for duration of exposure to the NICU (controls) . RESULTS: During a five-month period, C tropicalis was isolated from 29 blood cultures of six premature infants . The same organism also was isolated from fingernail samples taken from the ward housekeeper, who had a mild onychomycosis, and an asymptomatic nurse . Other potential reservoirs of C tropicalis were not identified among all the other infants or in the hospital environment . The six patients with C tropicalis fungemia were more likely to have received a larger number of antibiotics (4.0 versus 1.8, P < 0.001) and to have been subjected to a longer duration of total parenteral nutrition (TPN) therapy (8.5 versus 2.67 days, P = 0.004) than the controls . CONCLUSIONS: The risk of fungemia in this outbreak can be attributed to a larger number of antibiotics and a longer period of TPN administered to the patients . Analysis of events suggests that the outbreak may have been the result of cross-infection between staff and patients.

Am J Med Sci, 1993 Oct, 306(4), 225 - 32
Fungemia in patients with leukemia; Martino P et al.; A nine-year retrospective study on fungemia in patients with leukemia was conducted . A total of 79 episodes of fungemia in 77 patients with leukemia were documented . Candida parapsilosis fungemia was associated more frequently with the presence of a central venous line and to the use of parenteral nutrition than the other fungal species (p = 0.00026 and p = 0.01, respectively) . The same fungus was isolated from both blood and surveillance cultures in 95% of Candida albicans and in 89% of Candida tropicalis fungemia (p < 0.01 and p = 0.02, respectively) . The neutropenia and fungus colonization that resulted was associated significantly with the presence of invasive disease (p = 0.0024 and p = 0.0028, respectively) . Conversely, central venous catheterization and parenteral nutrition appeared to be associated with episodes without deep tissue invasion (p = 0.000037 and p = 0.001, respectively) . Invasive mycosis due to the fungus isolated from blood was documented in 51 patients with a mortality rate of 69%, whereas in 20 patients without invasive mycosis, mortality rate was 21% (p = 0.000059) . In patients with fungemia, related or unrelated to the presence of a central venous catheter, mortality was 24% and 64%, respectively (p = 0.00042) . Mortality was highest with C . tropicalis (p = 0.0017) and lowest with C . parapsilosis (p = 0.057) . Severe neutropenia (polymorphonuclears < 100/mmc) appeared associated with a higher mortality rate (p = 0.012), whereas the recovery of neutropenia was related adversely to a fatal outcome (p < 0.01) . With antifungal therapy, there was no statistically significant difference whether antifungal therapy was given or not.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1993 Sep 25, 268(27), 20143 - 7
Recombinant canditropsin, an extracellular aspartic protease from yeast Candida tropicalis . Escherichia coli expression, purification, zymogen activation, and enzymic properties; Lin X et al.; A cDNA fragment which encodes the zymogen of canditropsin, the extracellular aspartic protease from the yeast Candida tropicalis (Togni,G., Sanglard, D., Falchetto, R., and Monod, M . (1991) FEBS Lett . 286, 181-185) was cloned into a T7 expression vector for the synthesis of the recombinant zymogen in Escherichia coli . Recombinant canditropsinogen (Ctg), which was expressed as inclusion bodies in the cytosol of E . coli, was refolded by dialysis from an 8 M urea solution and purified to homogeneity using chromatographies on Sephacryl S-300 and on MonoQ columns . The purified Ctg was converted into canditropsin by either acid activation or trypsin conversion . The specificity of the resulting recombinant canditropsin toward polypeptide substrates is significantly different from other aspartic proteases . Canditropsin hydrolyzes oxidized insulin B chain between Ala-Leu and many other minor cleavage sites . Canditropsin also hydrolyzes keratin and collagen, which are components of connective tissues known to be hydrolyzed by canditropsin during Candida infections . Canditropsin was strongly inhibited by the universal aspartic protease inhibitor pepstatin (Ki = 1.75 x 10(-8) M) and inactivated by two aspartic protease inactivators, DAN and EPNP . Canditropsin is weakly inhibited by leupeptin and antipain, with an apparent Ki of 1.74 x 10(-4)M and 1.5 x 10(-5) M, respectively.

Biochim Biophys Acta, 1993 Sep 19, 1151(2), 175 - 85
Saccharomyces cerevisiae expression of exogenous vacuolar ATPase subunits B; Pan YX et al.; The precise function of subunit B of the vacuolar H(+)-ATPase class is unknown, but it is essential for proton pumping . We have previously reported the DNA sequence and predicted protein sequence of the vacuolar ATPase subunit B for Candida tropicalis (Gu, H.H., Gallagher, M.J., Rupkey, S . and Dean, G.E . (1990) Nucleic Acids Res . 18, 7446) . When the Candida gene was expressed in a Saccharomyce cerevisiae delta vat2 mutant from which the homologous gene had been deleted, viability and vacuolar acidification was restored to apparently wild-type levels . The predicted identity between these two proteins is 90% . We have searched for vacuolar ATPase subunits B from other species that might show a difference in function, when expressed in yeast, relative to the endogenous gene . We have cloned an apparently full-length 1.8-kb bovine subunit B cDNA from adrenal medulla that is about 1 kb shorter than the previously reported bovine brain cDNA (Puopolo, K., Kumamoto, C., Adachi, I., Magner, R . and Forgac, M . (1992) J . Biol . Chem . 267, 3696-3706; Nelson, R.D., Guo, X.L., Masood, K., Brown, D., Kalkbrenner, M . and Gluck, S . (1992) Proc . Natl . Acad . Sci . USA 89, 3541-3545), but nearly identical throughout the coding nucleotide and protein sequences; it is only 74% identical to the Saccharomyces subunit B protein sequence . Upon expression of this cDNA in two different delta vat2 deletion strains, the bovine cDNA restored function only partially, as judged by both viability at high pH and vacuolar acidification . Current work is aimed at determining which regions of the bovine protein require alteration in order to fully restore the delta vat2 strain to wild-type acidification, with the eventual goal of identifying interactive residues between subunit B and other proteins required for pump function.

Eur J Biochem, 1993 Sep 1, 216(2), 477 - 85
Probing the membrane topology of Candida tropicalis cytochrome P450; Sanglard D et al.; The membrane topology of two alkane-inducible cytochromes P450 from the yeast Candida tropicalis, alk1 and alk2, was tested by construction of fusion proteins with part of invertase and histidinol dehydrogenase (invHIS4C) and expression in a Saccharomyces cerevisiae his4 mutant . Depending on the localization of invHIS4C on the endoplasmic reticulum (ER) cytoplasmic or luminal side, the enzyme converts histidinol to histidine and allows the his4 yeast strain to grow on histidinol-supplemented medium . The N-terminal segments of alk1 and alk2 were fused to invHIS4C at three different locations that follow the first alk1 and alk2 transmembrane domains or a second putative transmembrane domain of alk1 . The combination of this in vivo assay with subcellular immunoprecipitations of the expressed fusion proteins allowed us to establish that both P450s contain only one transmembrane domain with their N-terminus located in the ER lumen . Deletions performed in these fusion proteins removing the first transmembrane domain of alk1 (delta TM) resulted in a less efficient targeting to the ER membrane but did not prevent their insertion in these membranes . Furthermore deletion of a negatively charged peptide preceding the first alk1 transmembrane domain (delta L) in an invHIS4C protein fused after this domain caused the N-terminal to have a positive net charge and to be oriented in the cytoplasm thus translocating the remaining protein into the ER lumen . The presence of the second hydrophobic segment, however, prevented the complete translocation of this fusion protein into the ER lumen . This study describes the first assessment of P450 membrane topology using an in vivo technique.

Crit Care Med, 1993 Sep, 21(9), 1324 - 7
Effect of extracorporeal membrane oxygenation on neutrophil function in neonates; DePuydt LE et al.; OBJECTIVE: To study the effect of long-term extracorporeal membrane oxygenation (ECMO) support on neutrophil function . DESIGN: A prospective, clinical investigation . SETTING: A pediatric intensive care unit . PATIENTS: Four groups of patients: ECMO group 1, newborns after 1 day of ECMO support (n = 10); ECMO group 2, newborns after 5 days of ECMO support (n = 6); group 3, normal newborns (n = 20); group 4, normal adults (n = 30) . INTERVENTIONS: Two mL of heparinized blood was obtained from patients in each group . A modification of the Smith and Rommel technique was used to measure neutrophil phagocytosis and killing utilizing live Candida tropicalis as the test organism . Neutrophils were incubated for 90 mins in normal adult serum with live Candida . Viability of Candida after phagocytosis was tested by vital fluorochrome staining . Phagocytic index (the number of neutrophils with intracytoplasmic Candida divided by the total neutrophils) and candidicidal ratio (neutrophils with dead Candida divided by total neutrophils with Candida) were determined daily . MEASUREMENTS AND MAIN RESULTS: Neutrophils from ECMO group 1 (day 1) and ECMO group 2 (day 5) patients had significantly higher phagocytosis indices (72.8 +/- 20 and 76 +/- 18) and candidicidal ratios (0.15 +/- 0.1 and 0.16 +/- 0.09) compared with neutrophils from group 3 patients (normal neonates) (64 +/- 7 and 0.06 +/- 0.04) . The phagocytosis indices were significantly lower in neutrophils from ECMO group 1 (day 1) and ECMO group 2 (day 5) patients compared with group 4 (adults) patients (86 +/- 9) . However, the candidicidal ratios in neutrophils from ECMO groups 1 and 2 (ECMO day 1 and day 5) patients were equal to that value in group 4 (adults) (0.10 +/- 0.04) . ECMO support for 5 days (ECMO group 2 vs . group 1) did not significantly change either the phagocytosis index or candidicidal ratio . CONCLUSIONS: Phagocytosis and intracellular killing by neutrophils of ECMO-supported neonates were significantly greater than those values found in normal newborns . ECMO support for 5 days produced no significant changes in neutrophil phagocytosis or killing.

J Perinatol, 1993 Sep-Oct, 13(5), 402 - 4
Treatment of Candida arthritis with flucytosine and amphotericin B; Weisse ME et al.; A premature infant with disseminated Candida tropicalis infection with arthritis and osteomyelitis is presented . Although the organism was susceptible to amphotericin B, synovial fluid cultures were still positive for yeast after 2 weeks of intravenous amphotericin B therapy . The addition of oral flucytosine (25 mg/kg four times daily) resulted in sterilization of the synovial fluid within 4 days . Simultaneous serum and synovial levels of flucytosine were 47.5 micrograms/ml and 39.6 micrograms/ml, respectively . This is the first documentation in the medical literature of intraarticular levels of flucytosine, and provides further rationale for the use of flucytosine in addition to amphotericin B in patients with Candida arthritis.

Antimicrob Agents Chemother, 1993 Sep, 37(9), 2030 - 2
Comparison of fluconazole and amphotericin B for treatment of experimental Candida endocarditis caused by non-C . albicans strains; Witt MD et al.; Amphotericin B and fluconazole were compared for the treatment of experimental Candida endocarditis caused by Candida tropicalis and C . parapsilosis . Rabbits received no therapy, amphotericin B (1 mg/kg of body weight per day intravenously), or fluconazole (100 mg/kg/day intraperitoneally) for either 11 or 21 days . Against both species, amphotericin B and fluconazole were equally effective overall; however, amphotericin B was more rapidly fungicidal than fluconazole in vivo against C . tropicalis.

Mycoses, 1993 Sep-Oct, 36(9-10), 299 - 303
Evaluation of two commercialized systems for the rapid identification of medically important yeasts; Quindos G et al.; A total of 77 recent clinical isolates of Candida albicans and other medically important yeasts were identified by two different commercial tests, Rapidec albicans (API-bioMerieux) and Fongiscreen 4H (Sanofi Diagnostics Pasteur), and conventional mycological methods . The strains were from 13 different species of yeasts and consisted of strains of 36 C . albicans, three of Candida famata, nine of Candida (Torulopsis) glabrata, five of Candida guilliermondii, two of Candida kefyr, three of Candida krusei, one of Candida lusitaniae, four of Cryptococcus neoformans, five of Candida parapsilosis, six of Candida tropicalis, one of Candida viswanathii, one of Rhodotorula rubra and one of Saccharomyces cerevisiae . According to the reactivity profiles of the isolates, identification was always correct with Fongiscreen 4H and was correct in 97.3% of the strains with Rapidec albicans . The latter test did not identify two C . albicans isolates that were correctly identified by Fongiscreen 4H . Both methods (97.3% correlation) were very useful for identification of C . albicans achieving the aim of their manufacturers . Additionally, Fongiscreen 4H was very useful for the identification of three other species of yeasts: C . glabrata, C . tropicalis and Cr . neoformans . The results of our study indicate that the accuracy of Rapidec albicans and Fongiscreen 4H is similar to that of the conventional methods used in this study for the identification of C . albicans . The same is true of Fongiscreen 4H in the identification of C . glabrata, C . tropicalis and Cr . neoformans . Both tests could be rapid and easy-to-perform tools in the clinical microbiology laboratory, but differences in cost must be taken into account.

FEMS Microbiol Lett, 1993 Aug 15, 112(1), 31 - 4
Presence of carnitine acetyltransferase in peroxisomes and in mitochondria of oleic acid-grown Saccharomyces cerevisiae; Atomi H et al.; Activity of carnitine acetyltransferase was detected in glucose- and oleic acid-grown Saccharomyces cerevisiae . Oleic acid-grown cells showed a ten-fold higher activity than glucose-grown cells . Subcellular fractionation of oleic acid-grown cells showed that carnitine acetyltransferase was present in peroxisomes, mitochondria, and cytosol . The results suggested the plausible presence of an 'acetylcarnitine shuttle' in this yeast, as in the case of an n-alkane-assimilating yeast, Candida tropicalis.

Clin Infect Dis, 1993 Aug, 17(2), 270 - 2
Hematogenous endophthalmitis due to Candida tropicalis: report of two cases and review; Cohen M et al.; Candida tropicalis is a well-documented pathogen affecting humans . There is limited clinical and experimental evidence that C . tropicalis causes hematogenous endophthalmitis . We report two cases of C . tropicalis endophthalmitis and review 12 cases reported in the literature . Clinical presentation was similar to that described for Candida albicans endophthalmitis . Therapy with amphotericin B, with or without flucytosine, resulted in resolution of the lesions except in one patient, for whom enucleation of the eye was necessary to control the infection . None of the patients were leukopenic.

J Clin Microbiol, 1993 Aug, 31(8), 2021 - 30
Comparison of restriction enzyme analysis versus pulsed-field gradient gel electrophoresis as a typing system for Torulopsis glabrata and Candida species other than C . albicans; Vazquez JA et al.; Candida species have recently emerged as important nosocomial pathogens . Because of the lack of a reliable system for detecting differences within the same species, little is known about the epidemiology of infection with Candida species . We describe a typing system for Torulopsis glabrata and the non-C . albicans Candida species that uses contour-clamped homogeneous electric field electrophoresis (CHEF), a version of pulsed-field gradient gel electrophoresis, and compared it with restriction enzyme analysis (REA) of genomic DNA . One hundred seventeen clinical isolates from 40 patients were evaluated . CHEF and REA were performed on each of the isolates, and the results of the two procedures were compared . The REA procedure revealed 8 different types of Candida lusitaniae, 20 of Torulopsis glabrata, 5 of Candida tropicalis, 3 of Candida parapsilosis, and 7 of Candida kefyr, whereas the CHEF method revealed 14 different types of C . lusitaniae, 16 of T . glabrata, 10 of C . tropicalis, 10 of C . parapsilosis, and 7 of C . kefyr . The CHEF technique yielded unique patterns of electrophoretic karyotypes that could be used to distinguish intraspecies variations . When compared with REA, CHEF demonstrated greater sensitivity in recognizing subtle strain-to-strain variations in most isolates and will be a useful epidemiologic tool for studying non-C . albicans Candida species and T . glabrata.

Am J Infect Control, 1993 Aug, 21(4), 189 - 95
Natural history of bloodstream infections in a burn patient population: the importance of candidemia; Ekenna O et al.; Because of a perceived increase in Candida bloodstream infections in our burn unit, we retrospectively reviewed all the microbiologic data and the medical records of 209 patients with burns admitted during a 42-month period . Twice weekly burn wound cultures demonstrated that Candida species were the tenth most frequently isolated organisms (69/191 patients, 36%) . Despite the low frequency of isolation from burn wounds, Candida species were the most common organisms found in blood cultures and urine cultures . Of 49 patients with positive blood cultures, 16 (33%) had clinically significant culture growth of yeasts: Candida albicans, 12; Candida parapsilosis, 2; Candida tropicalis, 1; and Torulopsis glabrata, 1 . Patients with candidemia were more likely than patients with blood culture growth of other organisms to have burn wound cultures that grew Candida (15/16 vs 21/33, p = 0.02), larger burns (61% vs 38%, p < 0.001), and death (63% vs 27%, p = 0.02) . Multivariate analysis demonstrated that the total number of blood cultures with microorganism growth and large burn size were the greatest independent risk factors for candidemia . These data demonstrate that yeasts are pathogens of major importance in patients with burns, suggesting that in patients with burns who have suspected sepsis and large burn injury or a previous bacteremia, strong consideration should be given to administration of amphotericin B initiation of empiric antibacterial therapy.

Jpn J Antibiot, 1993 Aug, 46(8), 647 - 53
{In vitro susceptibility to fluconazole of fungal strains freshly isolated from child patients with deep-seated mycoses}; Yamaguchi H et al.; The susceptibility to fluconazole (FLCZ) were evaluated for 40 strains of 8 yeast species and 4 strains of 3 Aspergillus species, all of which were isolated from clinical specimens during clinical trials of FLCZ in child patients performed from January 1991 to January 1993 in this country . The in vitro activity of FLCZ against yeast isolates and Aspergillus isolates were measured using the microdilution method with semisolid SAAMF (synthetic amino acid medium fungal) agar and the macrodilution method with Eagle MEM (minimum essential medium), respectively, using amphotericin B (AMPH) and flucytosine as the reference drugs . Out of all yeast species tested, Candida albicans and Candida tropicalis were the most susceptible to FLCZ with geometric mean (GM)-MICs of < 0.4 microgram/ml that were far less than the corresponding values for AMPH or flucytosine . On the other hand, FLCZ showed the lowest activity against Candida krusei and Candida glabrata with GM-MICs of approximately 10 micrograms/ml that were slightly higher than those for AMPH . The susceptibilities to FLCZ of Candida parapsilosis, Candida pelliculosa (anamorph of Hansenula anomala), Candida famata, and Trichosporon cutaneum were intermediate . Antifungal activities of FLCZ and the 2 reference drugs against aspergilli were determined on the basis of the amount of total protein recovered from drug-treated cultures relative to that from drug-free control cultures . Fifty percent and 90% inhibitory concentrations of FLCZ against 2 isolates of Aspergillus fumigatus were 6.25-25 micrograms/ml and 50 micrograms/ml, respectively . Against this and 2 other species of Aspergillus, AMPH appeared more highly active than FLCZ.

Diagn Microbiol Infect Dis, 1993 Jul, 17(1), 45 - 51
In vitro antimicrobial activity of tioconazole and its concentrations in vaginal fluids following topical (vagistat-1 6.5%) application; Jones RN et al.; In vitro assays demonstrated that clinical yeasts were significantly more inhibited by tioconazole (MIC50, < or = 0.5 microgram/ml) than by fluconazole (MIC50, 8 micrograms/ml) . Tioconazole also exhibited high potency against most molds (Alterneria spp . and Acremonium spp.) . All Candida tropicalis isolates had MICs of 8 micrograms/ml, four-fold greater than any other Candida spp . Generally Gram-negative bacteria were less susceptible to tioconazole . Moraxella catarrhalis (MIC90, 2 micrograms/ml) was the most susceptible Gram-negative species . Staphylococci and enterococci were the most susceptible to tioconazole Gram-positive species (MIC50s, 1-8 micrograms/ml) . Bacterial species associated with vaginosis . {Gardnerella vaginalis (MIC90, 16 micrograms/ml), Mobiluncus spp . (MIC90, 16 micrograms/ml) and Prevotella biviadisiens (MIC90, 64 micrograms/ml)} were inhibited by tioconazole . Isolates of Lactobacillus spp . were most resistant (MIC90, > or = 256 micrograms/ml) to tioconazole . Vaginal fluid levels of tioconazole (mean, 91.4 micrograms/ml) persisted above the MIC90 levels (1-64 micrograms/ml) for most fungal and bacterial pathogens for 72 h in 19 evaluable female human subjects receiving 300 mg tioconazole in an intravaginal ointment.

Biochim Biophys Acta, 1993 Jul 1, 1168(3), 280 - 4
Specific labeling of Candida tropicalis peroxisomal proteins with photoreactive fatty-acid derivatives; Mangroo D et al.; The labeling of Candida tropicalis peroxisomal proteins with photoreactive fatty-acid derivatives was investigated . Proteins having molecular masses of 70 kDa, 48 kDa and 15 kDa were labeled with 11-m-diazirinophenoxy-{11-3H}undecanoate while 11-m-diazirinophenoxy-{11-3H}undecanoyl-CoA labeled proteins of 70 kDa and 55 kDa . The 70 kDa protein labeled with both photoreactive probes was resolved into two bands by electrophoresis on a gradient polyacrylamide gel; immunoprecipitation with anti-fatty acyl-CoA oxidase showed that these proteins are fatty-acyl-CoA oxidases . In purified peroxisomal membranes, two proteins of 36 kDa and 25 kDa were labeled with the photoreactive fatty-acid probe, whereas very little labeling of the above proteins or other proteins was observed with the fatty-acyl-CoA probe . The photoaffinity labeling method described is, thus, clearly capable of identifying and distinguishing between proteins having an affinity for fatty acid or fatty-acyl-CoA . The labeling also identified a fatty-acid-binding site on the 16 kDa peroxisomal matrix protein as well as on two peroxisomal acyl-CoA oxidases . This approach thus provides a general means for the identification of fatty-acid metabolizing enzymes, as well as for the identification of fatty-acid-binding sites on known enzymes.

Leuk Lymphoma, 1993 Jul, 10(4-5), 369 - 76
Candida tropicalis infections in children with leukemia; Flynn PM et al.; The Candida species account for approximately three-fourths of fungal infections in patients with cancer . Although Candida albicans is the most frequent cause, C . tropicalis is increasingly implicated as an important pathogen . Over a 12 year period 19 children treated for leukemia at our institution developed C . tropicalis infections . We describe their clinical presentation, extent of fungal infection, treatment, and outcome . Fungemia without meningitis in 11 children was treated successfully, whereas C . tropicalis meningitis in 7 children was uniformly fatal . An additional patient had unsuspected, widespread infection detected at autopsy . Multiple sites, including the cerebrospinal fluid yielded C . tropicalis . Previously reported risk factors including neutropenia, broad-spectrum antibiotic usage, corticosteroid therapy, and total parenteral nutrition were observed in our cases . A high index of suspicion and the early use of aggressive antifungal therapy are critical to the successful management of C . tropicalis infections in children with leukemia.

Zhonghua Min Guo Xiao Er Ke Yi Xue Hui Za Zhi, 1993 Jul-Aug, 34(4), 257 - 63
Candida tropicalis fungemia in children with leukemia and lymphoma; Huang JL et al.; Five episodes of fungemias are described; all had occurred in children with leukemia or lymphoma between January 1, 1978 and December 31, 1990 . These fungemias comprised 3.4% of the total septicemias encountered during that period . Three episodes occurred during the induction phase and two during relapse . All patients had fever of varying degree and duration . In addition to steroids, all were receiving combination antibiotics before the fungemia had occurred . All patients had severe neutropenia lasting more than one week . Bacteremia preceded fungemia in four patients . Two episodes were diagnosed antemortem . The same species were isolated from other sites in three cases . Fever, chills and gastrointestinal symptoms were the most common clinical features; other symptoms included cough, dyspnea, oliguria and azotemia . One patient experienced skin lesion, dysphagia, hoarseness and hemiparesis . Only one patient survived . The prognosis from fungemia in leukemia and lymphoma patients is very poor . Empiric antifungal therapy is indicated in neutropenic patients who have recurrent or persistent fever despite one week of broad spectrum antibiotics . Early diagnosis and treatment will aid in improving the overall poor outcome of this disease.

Mycoses, 1993 Jul-Aug, 36(7-8), 247 - 53
Changes in the spectrum of fungal isolates: results from clinical specimens gathered in 1987/88 compared with those in 1991/92 in the University Hospital Göttingen, Germany; Borg-von Zepelin M et al.; In the University Hospital in Gottingen, the spectra of fungal species in clinical specimens of respiratory secretions, bronchial secretions and urine were compared over periods of 15 months (10/87 to 12/88 and 1/91 to 3/92) before and after the introduction of fluconazole . The following changes could be demonstrated: 1 . In all specimens analysed the number of Candida albicans isolates decreased, while the number of Candida tropicalis isolates remained almost unchanged . 2 . During the observation period the number of Candida glabrata isolates doubled . In 1991 C . glabrata was second to C . albicans as the most common of all fungal isolates, appearing in 8.6% of all specimens . 3 . The total number of Candida krusei isolates increased only slightly, but the rise in the number of isolates in bronchial secretions was statistically significant . 4 . The prevalence of rarely isolated Candida yeasts, such as Candida guilliermondii, Candida lipolytica and Candida kefyr, and Candida isolates which were not further differentiated increased . 5 . During the observation period the number of mixed cultures showed a fourfold increase . C . glabrata and C . krusei were associated in more than 75% of all isolates with C . albicans or C . tropicalis respectively . 6 . The number of mould isolates increased . These changes in the spectra of fungal isolates are discussed with respect to the broad therapeutic and prophylactic usage of fluconazole in the University Hospital of Gottingen.

Rev Clin Esp, 1993 Jun, 193(2), 70 - 2
{Usefulness of serology in the diagnosis of Candida tropicalis septicemia}; Quindos G et al.; In this study, we have evaluated the usefulness of eight serologic tests in the diagnosis of a Candida tropicalis septicaemia in a patient with acute myeloid leukaemia . Among the tests used, detection of mannose and candida antigen in serum allowed a high degree of suspicion of infection, earlier than the isolation of this species on mycological cultures from blood or oesophagic samples . Detection of anti-C . albicans antibodies was useless but low titres of anti-C . tropicalis antibodies were observed . In spite of early diagnosis and antifungal treatment, the complication of the disseminated candidiasis with a Pseudomonas aeruginosa septicaemia, concluded with the patient's exitus.

Bone Marrow Transplant, 1993 Jun, 11(6), 465 - 70
Systemic Candida infection in pediatric BM autotransplantation: clinical signs, outcome and prognosis; Besnard M et al.; Of the 393 children who underwent BM autotransplantation in the pediatric oncology unit of the Institut Gustave Roussy between February 1979 and September 1991, 14 (3.56%) developed disseminated Candida infection within 3 months . This incidence was far lower than in other published series . Eleven of the 14 patients recovered from the infection, giving a far higher survival rate (78%) than among adult BM transplant recipients (usually < 30%) . All 14 patients had four or more risk factors and persistent BM aplasia (median, 25 days); six had Candida tropicalis infection . Four cases of deep visceral involvement were documented, two of which were lethal . Clinical signs were relatively uniform and included secondary high-grade fever (> 40 degrees C) for 8 days; half the patients developed cardiovascular impairment, respiratory distress, neurological disturbances (altered consciousness or delirium) and severe diarrhoea, within as little as 10 days after transplantation . Blood cultures rapidly became positive after the onset of clinical signs and this permitted prompt treatment with a combination of amphotericin B and 5-fluorocytosine; in addition, central catheters were removed . Blood cultures became sterile within 4 days of treatment in 10 of the 14 cases . The generally favourable outcome in this series could be related to the young age of the patients, the absence of GVHD, the absence of total body irradiation in the conditioning regimen, and early antifungal treatment.

Gastrointest Endosc, 1993 May-Jun, 39(3), 413 - 5
Intraluminal fungal colonization of gastrostomy tubes; Gottlieb K et al.; Percutaneous endoscopic gastrostomy tubes are frequently colonized with fungal and bacterial organisms . This has not been previously reported . In our sample of 10 patients, nine percutaneous endoscopic gastrostomy tubes were colonized with fungi . This occurred as early as 1 week after placement . Candida tropicalis was isolated in five patients . It is hypothesized that a variety of fungi use components of the gastrostomy tube polymer, such as polymer additives, which contribute to the structural deterioration of the tube.

Medicine (Baltimore), 1993 May, 72(3), 143 - 50
Candida krusei fungemia . Report of 4 cases and review of the literature; Goldman M et al.; Candida krusei has recently been increasingly recognized as a pathogen in immunocompromised patients with malignancies . We report four immunocompromised patients with C . krusei fungemia and review the literature . Including our 4 cases, 62 cases of C . krusei fungemia were identified in the literature . Detailed information was available for only 25 patients . The clinical features of patients with C . krusei fungemia are similar to those reported for Candida tropicalis . Most patients were neutropenic and more than one half of the patients had received antifungal therapy and had evidence of gastrointestinal mucosal breakdown before the development of C . krusei fungemia . The overall mortality was 48% . Patients treated with regimens containing amphotericin B had improved survival over patients who received no therapy . Favorable response rates were higher in patients receiving high-dose amphotericin B or high-dose amphotericin B plus flucytosine when compared to patients treated with low-dose amphotericin B.

Arch Pathol Lab Med, 1993 May, 117(5), 521 - 3
Concurrent isolation of Candida krusei and Candida tropicalis from multiple blood cultures in a patient with acute leukemia; Sandin RL et al.; Reports of the concurrent isolation of more than one non-albicans species of Candida from blood cultures of immunocompromised patients with disseminated candidiasis are extremely infrequent . We report on the isolation of Candida krusei and Candida tropicalis from 17 blood cultures that were taken from a 67-year-old white man with a diagnosis of acute biphenotypic leukemia during a 2-week period of hospitalization for induction chemotherapy . Despite receiving high-dose amphotericin B throughout this period, the status of the patient worsened, and he experienced pancytopenia, hypernatremia, azotemia, and disseminated intravascular coagulation, which led to his death . Candida krusei and C tropicalis were isolated concurrently from 10 of the 17 blood cultures, while C krusei was the single isolate in three cultures and C tropicalis was isolated alone in four cultures . Each species manifested markedly different colonial morphological features . This case report serves to emphasize to microbiologists that they must exercise extreme suspicion when non-albicans species of Candida are isolated singly or concurrently from blood cultures in neutropenic patients, given the increasing clinical significance of these yeasts.

J Biol Chem, 1993 Apr 5, 268(10), 7372 - 81
Peptide splicing in the vacuolar ATPase subunit A from Candida tropicalis; Gu HH et al.; Subunit A of the vacuolar proton pump appears to be responsible for the ATP hydrolysis which is coupled to the pumping of protons into a variety of intracellular acid compartments, including the fungal vacuole . We report here the cloning and sequence determination of the gene encoding subunit A from Candida tropicalis . Southern blot hybridization analysis indicates that there is a single gene which encodes this protein . The gene contains a single intron at the extreme 5'-end of the coding region . The gene is predicted to encode a polypeptide of 1088 residues with a calculated molecular mass of 119,019 daltons, yet the mature polypeptide appears to be approximately 67 kDa, indicating that this protein probably undergoes the same sort of processing that is evidenced in the homologous protein from Saccharomyces cerevisiae in which an approximately 50-kDa polypeptide (the spacer) is spliced out of the mature protein . The Candida gene, with and without this middle portion, has been expressed in S . cerevisiae and found to restore a Saccharomyces subunit A deletion mutant (tfp1-delta 8) to apparently wild-type growth at pH 7.6, and normal vacuolar acidification . The peptide sequence of the two predicted mature ends is very similar to the sequences of the analogous proteins from Daucus carota, S . cerevisiae, and Neurospora crassa (60.5, 87.4, and 72.9% identity, respectively), but the middle portion bears only very limited homology with the Saccharomyces protein sequence . Processing of the gene product occurs in S . cerevisiae, Escherichia coli, and in rabbit reticulocyte-mediated in vitro translation, indicating that the excision is probably autocatalytic . The limited sequence identity seen between the Saccharomyces and Candida spacer domains may considerably narrow the functionally important regions responsible for the excision event.

J Clin Microbiol, 1993 Apr, 31(4), 904 - 10
Rapid, polymerase chain reaction-based identification assays for Candida species; Niesters HG et al.; Polymerase chain reaction (PCR) amplification of specific regions in the genomes of a variety of lower eukaryotes permits rapid identification of these microorganisms . First, on the basis of the presence of both constant and variable regions in the small subunit (ssu) rRNA, a nested PCR for direct identification of various Candida species can be designed . Amplification of the entire ssu rRNA gene and subsequent reamplification of variable sequences within the V4 domains of these PCR products were combined with direct sequencing . Restriction enzyme maps were made, and species-specific oligonucleotides for hybridization analysis were selected . Unequivocal discrimination of four of the major human pathogenic yeasts (Candida albicans, Candida glabrata, Candida tropicalis, and Candida krusei) is possible if a combination of these techniques is used . Second, by using oligonucleotides aimed at repeated sequences which occur at dispersed positions in the genomes of all eukaryotes, species-specific DNA fingerprints could be generated . This interrepeat PCR using genomic DNA as template proved to be an effective tool in Candida species typing . Both techniques described here can be extrapolated to the high-speed diagnostics of numerous other prokaryotic and eukaryotic pathogens.

Enferm Infecc Microbiol Clin, 1993 Mar, 11(3), 135 - 8
{Evaluation of a new automated system for the determination of antifungal sensitivity}; Lozano MC et al.; BACKGROUND: To evaluate a new semi-automatized system (ATB-Fungus, Bio Merieux, Spain) for antifungal in vitro susceptibility testing in yeast . METHODS: We studied the susceptibility of 91 yeast strains (22 Candida albicans, 18 Candida parapsilopsis, 18 Candida krusei, 17 Candida tropicalis, 11 Candida glabrata, 5 Candida guillermondii) against amphotericin B, flucytosine, ketoconazole, miconazole, econazole and nystatin, by means of "ATB-fungus" and agar diffusion and only against systemic usage antifungal agents by agar-dilution method . RESULTS: The correlation between "ATB-fungus" and diffusion method and agar dilution was 100% for amphotericin B, except for Candida krusei and C . tropicalis strains . This correlation rate was lower for fluocytosine and ketoconazole, without major differences between species . There is an absolute concordance between the system under evaluation and agar diffusion method regarding nystatin . For the two azole drugs remaining (miconazole and econazole) the correlation found for Candida glabrata was 100% and for C . parapsilopsis 95% with agar diffusion method . Candida glabrata was the single species that showed better correlation with all three methods, while major differences were seen with C . krusei and C . tropicalis . CONCLUSIONS: Although "ATB-fungus" seems to be a nearly well standardized method for antifungal susceptibility testing, more studies are required for assessing its reproducibility and also to determine when, how and for which antifungal agents would be a valid procedure.

J Clin Microbiol, 1993 Mar, 31(3), 518 - 23
Diagnosis of invasive candidiasis by a dot immunobinding assay for Candida antigen detection; Reboli AC; A dot immunobinding assay which uses a polyclonal rabbit anti-Candida immunoglobulin G as the primary antibody and colloidal gold coated with goat anti-rabbit immunoglobulin G as the secondary antibody for the detection of Candida cytoplasmic antigens is described . It was able to detect as little as 1 ng of total Candida protein per ml when a cytoplasmic extract of Candida albicans was seeded into buffer and 10 ng/ml when the same extract was seeded into pooled human serum . Serial serum samples from four groups of patients were assayed for Candida antigen: (i) 22 patients with candidemia, (ii) 16 patients at high risk for invasive candidiasis, (iii) 3 patients with other deep mycoses, and (iv) 50 hospitalized patients at low risk for serious Candida infection . Of the 22 candidemic patients, 19 had invasive candidiasis and 3 had transient candidemia . Antigenemia was detected in 16 of the 19 patients with invasive candidiasis (including patients with C . albicans, Candida tropicalis, Candida glabrata, Candida krusei, and Candida parapsilosis) and in 4 of 16 patients at high risk for invasive candidiasis . There was no detectable antigen in 12 high-risk control patients, 3 patients with transient candidemia, 3 patients with other deep mycoses, and 50 relatively low-risk patients . The sensitivity for detecting invasive disease in candidemic patients and specificity for all patients studied were 84.2 and 94.4%, respectively . The positive predictive value was 80%; the negative predictive value was 95.7% . The sensitivity for neutropenic patients with invasive disease was 85.7% . This assay is rapid and accurate and appears to be useful in identifying candidemic patients with invasive candidiasis.

Clin Infect Dis, 1993 Mar, 16(3), 377 - 83
Comparison of pulsed-field gel electrophoresis with isoenzyme profiles as a typing system for Candida tropicalis; Doebbeling BN et al.; Candida species are important nosocomial pathogens, particularly in immunocompromised and critically ill patients . A variety of methods have been used to differentiate strains, but an optimal system has not been established . We compared methods for typing a panel of nine related isolates of Candida tropicalis from an outbreak of sternal wound infections as well as four unrelated control isolates of this species . (The genetic relationships of the nine isolates in the panel had been confirmed previously by restriction fragment analysis.) Typing was undertaken without knowledge of an isolate's origin . Karyotyping by contour-clamped homogeneous electric field (CHEF) gel electrophoresis failed to distinguish between outbreak and control isolates . However, when chromosome-sized DNA was digested with SfiI, EagI, SacII, or NaeI and the fragments were separated by CHEF electrophoresis, the outbreak isolates were readily identified . The isoenzyme profiles of the outbreak isolates were identical and were distinctly different from those of the control isolates . While both isoenzyme profiles and the modified CHEF procedure were discriminatory, the latter is recommended as a relatively convenient and reproducible technique for comparison of types of C . tropicalis.

J Am Vet Med Assoc, 1993 Feb 1, 202(3), 413 - 5
Ventral dermatitis and vasculitis in a calf with bovine leukocyte adhesion deficiency; Ackermann MR et al.; Dermatitis and vasculitis of the skin covering the sternum of a calf with bovine leukocyte adhesion deficiency was observed . The calf had been conceived through artificial insemination, delivered by cesarean section, and placed in a gnotobiotic isolator . Dermatitis was noticed at 54 days of age and was not responsive to antibiotic or ivermectin treatment . Proteus sp, Enterobacter sp, and Candida tropicalis were isolated from skin specimens . The lesion was characterized by lymphoplasmacytic and histiocytic inflammation with vasculitis and thrombosis.

J Med Vet Mycol, 1993, 31(1), 1 - 15
Characterization of two monoclonal antibodies against secretory proteinase of Candida tropicalis DSM 4238; Borg-von Zepelin M et al.; Two murine IgM monoclonal antibodies (mAb; MT1 and MT2), which were produced against the secretory aspartic proteinase of Candida tropicalis DSM 4238, are described . Both antibodies reacted with the native and denatured conformations of the homologous proteinase antigen but showed different patterns of reactivity with other related proteinases (Candida albicans CBS 2730, serotype A; C . albicans ATCC 48867, serotype B; Candida parapsilosis DSM 4237) and with porcine pepsin . Neither of the antibodies inhibited the proteolytic activity of the homologous enzyme . MT1 also reacted with mannoproteins of C . tropicalis DSM 4238 and C . albicans CBS 2730 and immunofluorescence revealed that this antibody bound to the surface of blastoconidia and pseudomycelia of these two Candida species . A reaction with blastoconidia only was observed with C . albicans serotype B . MT1 also reacted weakly with Candida guilliermondii, but not with C . parapsilosis, Candida glabrata, Candida krusei or Candida kefyr . MT2 did not bind to fungal surfaces . Preliminary experiments suggested that mAb MT1 may recognize a carbohydrate epitope, while MT2 binds to an epitope consisting of the protein part of the enzyme . The two antibodies were used in an ELISA for the detection of proteinase antigen . ELISA with MT1 or MT2 as coating antibodies and a specific protein epitope recognizing mAb-biotin conjugate was able to detect 4 ng ml-1 of antigen . Trials with 26 sera from fungemic patients and 14 sera from controls suggest that MT2 is of potential value in antigen-directed serodiagnosis.

Urologe A, 1993 Jan, 32(1), 53 - 5
{Gas-forming mycosis of the kidney}; Vorreuther R et al.; We report on the case of a 22-year-old heroin-dependent diabetic woman . She presented with a gas-forming fungal infection of the kidney caused by Candida tropicalis . The patient was successfully treated by conservative means . Problems in the diagnosis and therapy of this rare disease are discussed.

Mikrobiol Zh, 1993 Jan-Feb, 55(1), 28 - 37
Alternation of exo- and endotrophy during the mitotic cycle of the yeast cells; Ivanov VN et al.; The utilization of the intracellular and extracellular sources of carbon and energy during the mitotic cycle of yeasts Saccharomyces cerevisiae, Kluyveromyces marxianus, Candida boidinii, Candida tropicalis has been studied . Increase in the consumption rate of carbon and energy sources and in the exogenous respiration rate at G1- and G2-phases of the mitotic cycle is shown . The rate of the endogenous respiration of the cells at these phases decreased . The hypothesis has been proposed that during the mitotic cycle of the yeast cell the regular alternation of exotrophy (the utilization of the extracellular carbon and energy sources by a cell) and endotrophy (the process of the utilization of the intracellular carbon and energy sources by a cell) occurs . It is possible to reveal the exotrophic cells by the cytological method which is based on the calculation of dead cells after incubation of the yeast suspension in amyl alcohol solution . This method has revealed that exotrophic and endotrophic processes do not predominate one over another but alternate at the mitotic cycle . Exotrophy and endotrophy are phase-specific processes . The G1- and G2-phases are exotrophic processes, phases S and M are endotrophic ones.

Antimicrob Agents Chemother, 1993 Jan, 37(1), 39 - 45
Multicenter evaluation of a broth macrodilution antifungal susceptibility test for yeasts; Fromtling RA et al.; Thirteen laboratories collaborated to optimize interlaboratory agreement of results of a broth macrodilution procedure for testing three classes of antifungal drugs against pathogenic yeasts . The activities of amphotericin B, flucytosine, and ketoconazole were tested against 100 coded isolates of Candida albicans, Candida tropicalis, Candida parapsilosis, Candida lusitaniae, Torulopsis (Candida) glabrata, and Cryptococcus neoformans . Two starting yeast inoculum sizes (5 x 10(4) and 2.5 x 10(3) cells per ml) were compared, and readings were taken after 24 and 48 h of incubation . All other test conditions were standardized . The resultant turbidities in all tubes were estimated visually on a scale from 0 to 4+ turbidity, and MIC-0, MIC-1, and MIC-2 were defined as the lowest drug concentrations that reduced growth to 0, 1+, or 2+ turbidity, respectively . For flucytosine, agreement among laboratories varied between 57 and 87% for different inocula, times of incubation, and end point criteria . Agreement was maximized (85%) when the lower inoculum was incubated for 2 days and the MICs were defined as 1+ turbidity or less . For amphotericin B, variations in test conditions produced much smaller differences in interlaboratory agreement . For ketoconazole, interlaboratory agreement was poorer by all end point criteria . However, MIC-2 endpoints distinguished T . glabrata as resistant compared with the other species . Overall, the studies indicated that readings from the lower inoculum obtained on the second day of reading result in the greatest interlaboratory agreement . In combination with data from previous multicenter studies (National Committee for Clinical Laboratory Standards, Antifungal Susceptibility Testing: Committee Report, Vol . 5, No . 17, 1988; M . A . Pfaller, L . Burmeister, M . S . Bartlett, and M . G . Rinaldi, J . Clin . Microbiol . 26:1437-1441, 1988; M . A . Pfaller, M . G . Rinaldi, J . N . Galgiani, M . S . Bartlett, B.A . Body, A . Espinel-Ingroff, R.A . Fromtling, G.S . Hall, C.E . Hughes, F . C . Odds, and A . M . SUgar, J . Clin . Microbiol . 34:1648-1654, 1990), these findings will be used by the National Committee for Clinical Laboratory Standards to develop a standardized method for in vitro antifungal susceptibility testing for yeasts.

Leuk Lymphoma, 1993, 11 Suppl 2, 127 - 35
What's new in fungal infection in leukemic patients; Bodey GP; Fungal infections have become an important cause of mortality in patients with hematological malignancies . In recent years, fungi such as Candida tropicalis, Aspergillus spp, Fusarium spp and Trichosporon spp have emerged as important pathogens . Amphotericin B remains the antifungal agent with the broadest spectrum of activity, although some of the newer pathogens may be resistant . The administration of this drug in lipid vehicles reduces the toxicities, permitting the administration of higher doses that may be more effective . The new agents, fluconazole and itraconazole, have activity against some fungal pathogens, although their role in therapy has not been fully determined . Fluconazole has been shown to be effective for prophylaxis of Candida infections.

Eur J Biochem, 1992 Dec 15, 210(3), 999 - 1005
Peroxisomal acetoacetyl-CoA thiolase of an n-alkane-utilizing yeast, Candida tropicalis; Kurihara T et al.; Two genes encoding acetoacetyl-CoA thiolase (thiolase I; EC 2.3.1.9), whose localization in peroxisomes was first found with an n-alkane-utilizing yeast, Candida tropicalis, were isolated from the lambda EMBL3 genomic DNA library prepared from the yeast genomic DNA . Nucleotide sequence analysis revealed that both genes contained open reading frames of 1209 bp corresponding to 403 amino acid residues with methionine at the N-terminus, which were named as thiolase IA and thiolase IB . The calculated molecular masses were 41,898 Da for thiolase IA and 41,930 Da for thiolase IB . These values were in good agreement with the subunit mass of the enzyme purified from yeast peroxisomes (41 kDa) . There was an extremely high similarity between these two genes (96% of nucleotides in the coding regions and 98% of amino acids deduced) . From the amino acid sequence analysis of the purified peroxisomal enzyme, it was shown that thiolase IA and thiolase IB were expressed in peroxisomes at an almost equal level . Both showed similarity to other thiolases, especially to Saccharomyces uvarum cytosolic acetoacetyl-CoA thiolase (65% amino acids of thiolase IA and 64% of thiolase IB were identical with this thiolase) . Considering the evolution of thiolases, the C . tropicalis thiolases and S . uvarum cytosolic acetoacetyl-CoA thiolase are supposed to have a common origin . It was noticeable that the carboxyl-terminal regions of thiolases IA and IB contained a putative peroxisomal targeting signal, -Ala-Lys-Leu-COOH, unlike those of other thiolases reported hitherto.

DNA Cell Biol, 1992 Dec, 11(10), 767 - 80
Identification and characterization of additional members of the cytochrome P450 multigene family CYP52 of Candida tropicalis; Seghezzi W et al.; Using different DNA probes from the first two previously described alkane-inducible cytochrome P450 genes of the Candida tropicalis CYP52 gene family, we isolated five independent additional members by screening a genomic library under low-stringency conditions . These genes are not allelic variants and, when taken gogether, constitute the largest gene family known in this organism . The seven members of this gene family are located on four different chromosomes and four of them are tandemly arranged on the C . tropicalis genome . The products of the seven genes, alk1 to alk7, were compared to each other and revealed a high degree of divergence: the two most diverged proteins exhibit a sequence identity of only 32% . Six of the seven genes were shown to be induced by a variety of different aliphatic carbon sources but repressed when the organism was grown on glucose . Three of the five additional CYP52 genes could be successfully expressed in Saccharomyces cerevisiae and display different substrate specificities in in vitro assays with model substrates: alk2 and alk3 exhibit a strong preference for hexadecane, while alk4 and alk5 preferentially hydroxylate lauric acid.

J Clin Microbiol, 1992 Dec, 30(12), 3132 - 7
Detection of serum Candida antigens by enzyme-linked immunosorbent assay and a latex agglutination test with anti-Candida albicans and anti-Candida krusei antibodies; Fujita S et al.; Serum samples from 197 patients with and without candidiasis were assayed for Candida albicans mannan and Candida krusei mannan by an enzyme-linked immunosorbent assay (ELISA) and a latex agglutination test (LA) and for D-arabinitol by the enzymatic fluorometric method . Of the 43 patients positive for C . albicans mannan (> or = 0.2 ng/ml), 34 were infected with C . albicans and 9 were infected with Candida tropicalis . C . krusei mannan (> or = 0.3 ng/ml) was detected in 10 patients infected with Candida parapsilosis, 2 patients infected with Candida guilliermondii, and 1 patient infected with C . krusei . With both anti-C . albicans antibodies and anti-C . krusei antibodies, the sensitivities of ELISA and LA for detection of invasive candidiasis (58 patients) were 74 and 38%, respectively . No false-positive reactions were observed by the ELISA or the LA . The sensitivity and specificity of the D-arabinitol/creatinine ratio (> or = 1.5 mumol/mg) to invasive candidiasis were 50 and 91%, respectively . The ELISA with antibodies against both C . albicans and C . krusei may be useful in diagnosing invasive candidiasis caused by medically important Candida strains excluding Candida glabrata.

J Biochem (Tokyo), 1992 Dec, 112(6), 845 - 8
Physiological roles of acetoacetyl-CoA thiolase in n-alkane-utilizable yeast, Candida tropicalis: possible contribution to alkane degradation and sterol biosynthesis; Kurihara T et al.; The presence of two types of thiolases, acetoacetyl-CoA thiolase and 3-ketoacyl-CoA thiolase, was demonstrated in peroxisomes of n-alkane-grown Candida tropicalis {Kurihara, T., Ueda, M., & Tanaka, A . (1989) J . Biochem . 106, 474-478}, while acetoacetyl-CoA thiolase was also shown to be present in cytosol . The activity of the enzyme in cytosol was constant irrespective of culture conditions, while the peroxisomal enzyme was inducibly synthesized in the alkane-grown yeast cells . These results indicate that peroxisomal acetoacetyl-CoA thiolase participates in alkane degradation, while the cytosolic enzyme is associated with other fundamental metabolic processes, probably sterol biosynthesis, because this enzyme can catalyze the first step of the sterol biosynthesis . 3-Hydroxy-3-methylglutaryl (HMG)-CoA reductase, a key regulatory enzyme of sterol biosynthesis, was found to be localized exclusively in microsomes of the alkane-grown yeast cells . These results suggest that yeast peroxisomes do not contribute to sterol biosynthesis, unlike the case of mammalian cells.

Methods Find Exp Clin Pharmacol, 1992 Dec, 14(10), 753 - 8
Effect of amphotericin B alone or in combination with rifampicin on phagocytosis of Candida species by human polymorphonuclear leukocytes; Schaumann RF et al.; The influence of amphotericin B and rifampicin alone and in combination on the killing of Candida albicans, Candida tropicalis and Cryptococcus neoformans by human polymorphonuclear leukocytes was investigated in vitro . Granulocytes were harvested from the blood of healthy volunteers and resuspended with the above mentioned fungi . After 6 hours, 81.5% of C . albicans, 97% of C . tropicalis but only 34% of C . neoformans were killed . The activity of amphotericin B against C . albicans and C . tropicalis was better in granulocyte-free medium . A synergism between amphotericin B and rifampicin towards Candida could be detected . It was also demonstrated in the presence of polymorphonuclear leukocytes with a factor > 1 log in reduction of colony count . However, whether granulocytes were present or not, rifampicin (4 mcg/mL) alone did not exert any influence on growth of Candida.

Yeast, 1992 Dec, 8(12), 1065 - 75
DNA transformations of Candida tropicalis with replicating and integrative vectors; Sanglard D et al.; The alkane-assimilating yeast Candida tropicalis was used as a host for DNA transformations . A stable ade2 mutant (Ha900) obtained by UV-mutagenesis was used as a recipient for different vectors carrying selectable markers . A first vector, pMK16, that was developed for the transformation of C . albicans and carries an ADE2 gene marker and a Candida autonomously replicating sequence (CARS) element promoting autonomous replication, was compatible for transforming Ha900 . Two transformant types were observed: (i) pink transformants which easily lose pMK16 under non-selective growth conditions; (ii) white transformants, in which the same plasmid exhibited a higher mitotic stability . In both cases pMK16 could be rescued from these cells in Escherichia coli . A second vector, pADE2, containing the isolated C . tropicalis ADE2, gene, was used to transform Ha900 . This vector integrated in the yeast genome at homologous sites of the ade2 locus . Different integration types were observed at one or both ade2 alleles in single or in tandem repeats.

Rev Rhum Mal Osteoartic, 1992 Nov 30, 59(11), 732 - 4
{Candida tropicalis arthritis of the shoulder associated with articular chondrocalcinosis}; Cerf I et al.; A case of infection of the scapulohumeral joint following articular infiltrations is reported in a female patient with alcohol-related cirrhosis of the liver . Joint destruction occurred despite appropriate treatment with fluconazole . Candida tropicalis joint infections are seen mainly in immunocompromised hosts and usually cause little destruction of the joint . Most are the result of dissemination via the bloodstream . Preexisting chondrocalcinosis in the patient reported here may have been a contributing factor to the development of extensive joint destruction.

J Urol, 1992 Nov, 148(5), 1536 - 8
Prostatic abscess due to Candida tropicalis in a nonacquired immunodeficiency syndrome patient; Yu S et al.; We report a case of prostatic abscess due to Candida tropicalis in a nonacquired immunodeficiency syndrome patient with diabetes . The diagnosis and management are discussed, and the literature is reviewed.

J Bacteriol, 1992 Nov, 174(22), 7379 - 84
Lysine biosynthesis in selected pathogenic fungi: characterization of lysine auxotrophs and the cloned LYS1 gene of Candida albicans; Garrad RC et al.; The alpha-aminoadipate pathway for the biosynthesis of lysine is present only in fungi and euglena . Until now, this unique metabolic pathway has never been investigated in the opportunistic fungal pathogens Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus . Five of the eight enzymes (homocitrate synthase, homoisocitrate dehydrogenase, alpha-aminoadipate reductase, saccharopine reductase, and saccharopine dehydrogenase) of the alpha-aminoadipate pathway and glucose-6-phosphate dehydrogenase, a glycolytic enzyme used as a control, were demonstrated in wild-type cells of these organisms . All enzymes were present in Saccharomyces cerevisiae and the pathogenic organisms except C . neoformans 32608 serotype C, which exhibited no saccharopine reductase activity . The levels of enzyme activity varied considerably from strain to strain . Variation among organisms was also observed for the control enzyme . Among the pathogens, C . albicans exhibited much higher homocitrate synthase, homoisocitrate dehydrogenase, and alpha-aminoadipate reductase activities . Seven lysine auxotrophs of C . albicans and one of Candida tropicalis were characterized biochemically to determine the biochemical blocks and gene-enzyme relationships . Growth responses to alpha-aminoadipate- and lysine-supplemented media, accumulation of alpha-aminoadipate semialdehyde, and the lack of enzyme activity revealed that five of the mutants (WA104, WA153, WC7-1-3, WD1-31-2, and A5155) were blocked at the alpha-aminoadipate reductase step, two (STN57 and WD1-3-6) were blocked at the saccharopine dehydrogenase step, and the C . tropicalis mutant (X-16) was blocked at the saccharopine reductase step . The cloned LYS1 gene of C . albicans in the recombinant plasmid YpB1078 complemented saccharopine dehydrogenase (lys1) mutants of S . cerevisiae and C . albicans . The Lys1+ transformed strains exhibited significant saccharopine dehydrogenase activity in comparison with untransformed mutants . The cloned LYS1 gene has been localized on a 1.8-kb HindIII DNA insert of the recombinant plasmid YpB1041RG1 . These results established the gene-enzyme relationship in the second half of the alpha-aminoadipate pathway . The presence of this unique pathway in the pathogenic fungi could be useful for their rapid detection and control.

J Bacteriol, 1992 Nov, 174(22), 7112 - 20
Phenol hydroxylase from Trichosporon cutaneum: gene cloning, sequence analysis, and functional expression in Escherichia coli; Kalin M et al.; A cDNA clone encoding phenol hydroxylase from the soil yeast Trichosporon cutaneum was isolated and characterized . The clone was identified by hybridization screening of a bacteriophage lambda ZAP-based cDNA library with an oligonucleotide probe which corresponded to the N-terminal amino acid sequence of the purified enzyme . The cDNA encodes a protein consisting of 664 amino acids . Amino acid sequences of a number of peptides obtained by Edman degradation of various cleavage products of the purified enzyme were identified in the cDNA-derived sequence . The phenol hydroxylase cDNA was expressed in Escherichia coli to yield high levels of active enzyme . The E . coli-derived phenol hydroxylase is very similar to the T . cutaneum enzyme with respect to the range of substrates acted upon, inhibition by excess phenol, and the order of magnitude of kinetic parameters in the overall reaction . Southern blot analysis revealed the presence of phenol hydroxylase gene-related sequences in a number of T . cutaneum and Trichosporon beigelii strains and in Cryptococcus elinovii but not in Trichosporon pullulans, Trichosporon penicillatum, or Candida tropicalis.

Infect Immun, 1992 Nov, 60(11), 4907 - 14
Gastrointestinal colonization and systemic dissemination by Candida albicans and Candida tropicalis in intact and immunocompromised mice; de Repentigny L et al.; Gastrointestinal colonization and systemic dissemination by Candida albicans and Candida tropicalis were compared in intact and immunocompromised mice . Five-day-old CFW mice were inoculated by the oral-intragastric route with 1.0 x 10(7) CFU of two C . albicans and two C . tropicalis strains isolated from the blood of patients with acute leukemia and with C . albicans 4918 and its cerulenin-resistant mutant 4918-10 . C . albicans and C . tropicalis spread to the lungs, liver, and kidneys within 30 min postinoculation, and organ CFU of the two species were comparable over the following 10 days . Close association of blastoconidia with the villous surface of the small intestine resulted in lysis of microvilli and then progressive invasion of villi . Blastoconidia within villi were surrounded by a conspicuous zone of clearing . Persistent colonization of the small and large intestines by C . albicans blood isolates and strains 4918 and 4918-10 was similar for 31 days after inoculation, but consistently exceeded that of C . tropicalis . In mice colonized with C . albicans, immunosuppression with cortisone acetate and cyclophosphamide on days 30 and 33 after inoculation increased stomach CFU 40- to 370-fold and intestinal CFU 30- to 80-fold . In contrast, persistent colonization by C . tropicalis was undetectable before immunosuppression and only became apparent after treatment . C . albicans disseminated more frequently and with higher organ CFU than C . tropicalis . Despite this fact, 20% of mice infected with C . tropicalis died, compared with 4% infected with C . albicans blood isolates . Indirect immunofluorescence revealed penetrative growth by Candida hyphae exclusively in the mucosa and submucosa of the stomach from immunosuppressed, persistently colonized mice . Taken together, the data indicate that C . tropicalis appears to be more virulent than C . albicans and that factors responsible for gastrointestinal colonization, systemic dissemination, and mortality in immunocompromised mice may not be identical.

Mycoses, 1992 Nov-Dec, 35(11-12), 371 - 4
Dermatomycoses among industrial workers in Cross River State, Nigeria; Akpata LE et al.; A survey of dermatomycoses was carried out amongst industrial workers in three different factories during the period 1987-1988 . A total of 194 workers were screened, out of which 54 proved to be mycologically positive by microscopy and/or culture . Incidence was apparently highest amongst workers in a cement factory (Calcemco, 33.3%) followed by those in a wood factory (Seromwood, 30.8%) and a rubber factory (CREL, 26.2%) . Pityriasis versicolor was the predominant clinical type of dermatomycosis, followed by tinea pedis . A total of 51 fungal organisms were identified . Malassezia furfur was the most prevalent causative agent (74.4%) followed by Trichophyton soudanense (5.8%), T . rubrum (3.9%) and Epidermophyton floccosum (3.9%) . Other species identified were one isolate each of T . tonsurans, T . mentagrophytes, T . verrucosum, Candida tropicalis, Candida spec . and Geotrichum candidum.

Mikrobiologiia, 1992 Nov-Dec, 61(6), 975 - 80
{Penetration of pancreatic DNase into Candida tropicalis cells}; Kupriianova-Ashina FG et al.; Permeability of Candida tropicalis cells for exogenic DNAse was studied by a cytochemical method . The enzyme were shown to penetrate yeast outer membrane and cell wall after a 20 minute incubation period when incubated together with cells at the beginning of the stationary phase.

Mikrobiologiia, 1992 Nov-Dec, 61(6), 969 - 74
{Effect of exogenous ribonucleases on the propagation of Candida tropicalis yeast}; Kolpakov AI et al.; The effect of exogenic microbial and tissue RNAses on the reproduction of Candida tropicalis yeasts were studied . Catalytically active and inactive forms of ribonucleases were shown to stimulate yeast organism reproduction . Using immunochemical and radioisotopic methods, it was found that RNAses penetrated into yeast cell . The correlation between yeast cell penetration by exogenic enzymes and the physiological state of the cells was revealed . The activation of the chromatin associated RNAse-polymerase under the action of the catalytically active binase was observed in in vitro experiments . The mechanisms of the stimulatory effect of catalytically active and inactive RNAses on growth and reproduction of C . tropicalis are discussed.

Clin Invest Med, 1992 Oct, 15(5), 434 - 9
The comparative efficacy of cilofungin, fluconazole and amphotericin B in disseminated Candida tropicalis infection in neutropenic mice; Fong IW et al.; There is insufficient in vivo data on the efficacy of new antifungal agents against invasive Candida tropicalis infection . Disseminated infection with Candida tropicalis in neutropenic mice was treated with cilofungin, fluconazole, or amphotericin B intraperitoneally, and compared to untreated controls . Early survival rates at the end of treatment (day 10) were similar for amphotericin B (97.5%) and fluconazole (100%), and superior to cilofungin (62.6%) which was better than no treatment (0%) . Late survival rates (day 31) were highest for amphotericin B (95%), and significantly lower for cilofungin (48.7%) and fluconazole (43.9%), p = 0.0001 . Rates of sterilization of the lung, liver, and spleen were high in survivors for all regimens (85.1-100%) but lower for the kidneys: fluconazole, 21.3%; amphotericin B, 39.3%; and cilofungin, 65.5% . Amphotericin B was the most effective agent in this study of disseminated Candida tropicalis (C . tropicalis) infection.

J Clin Microbiol, 1992 Oct, 30(10), 2748 - 52
Colony morphotype on Sabouraud-triphenyltetrazolium agar: a simple and inexpensive method for Candida subspecies discrimination; Quindos G et al.; A new method of Candida subspecies discrimination on Sabouraud-triphenyltetrazolium agar is reported . Five hundred sixty-two strains of Candida and Torulopsis glabrata, previously identified by conventional mycological methods, were studied . Each strain received a three-letter code and a number based on its colonial morphology . Sixteen morphotypes were found for Candida albicans, 6 were found for Candida parapsilosis, 4 were found for both Candida guilliermondii and Candida krusei, and 12 were found for Candida tropicalis . None of the 56 T . glabrata strains studied grew on this agar . A reproducibility of 95% was found for C . albicans . The simplicity and low cost could make this method useful for typing Candida spp.

Clin Infect Dis, 1992 Sep, 15(3), 414 - 21
Candidemia in a tertiary care hospital: epidemiology, risk factors, and predictors of mortality; Fraser VJ et al.; Demographic information, risk factors, therapy, and outcome for all patients who had candidemia at Barnes Hospital, St . Louis, between 1 September 1988 and 1 September 1989 were retrospectively reviewed . One hundred six candidemic patients were identified, representing 0.5% of all medical and surgical discharges and 0.33% of total patient discharges . These percentages represent a 20-fold increase in the incidence of candidemia at our hospital in comparison with that during 1976-1979 . Candida albicans was the most frequently isolated species (63%), followed by Candida tropicalis (17%), Candida glabrata (13%), Candida parapsilosis (6.5%), and Candida krusei (0.9%) . Overall mortality was 57%, and 14 (23%) of 60 deaths occurred within 48 hours of the detection of candidemia . Mortality was associated with higher APACHE II scores (25 for nonsurvivors vs . 16 for survivors; P = .0001), the presence of a rapidly fatal underlying illness (P = .0009), and sustained positivity of blood cultures (P = .02) . In cases of sustained candidemia, the isolation of non-albicans Candida species also correlated with increased mortality (8 of 8 vs . 10 of 21; P = .005) . Thirty candidemic patients (28%) did not receive any antifungal therapy, and 19 (63%) of these untreated patients died . Eleven untreated patients (37%) survived without sequelae . There has been a marked increase in the incidence of candidemia in our institution that is associated with a high overall mortality . Candidemia lasting less than 24 hours was associated with a lower mortality than was that of longer duration . Severity of illness and duration of candidemia should be used as stratifying factors in prospective studies to determine optimum therapy.

Monatsschr Kinderheilkd, 1992 Sep, 140(9), 633 - 8
{Candida infections in premature infants weighing less than 1,500 g . Mucocutaneous colonization and incidence of systemic infections}; Harms K et al.; BACKGROUND: An increasing incidence of systemic candidiasis has been reported in low birth weight infants requiring intensive care . We have retrospectively analyzed mucocutaneous Candida-colonization and infection rate in 422 preterm infants with a birthweight < 1,500 g . METHODS: All infants were treated at the NICU, University of Gottingen, from 1/1985-5/1991 . 359 neonates (85%) were on mechanical ventilation, no prophylactic antimycotic regimen was applied . Mucocutaneous swabs and cultures from various anatomic sites were regularly obtained from all infants . RESULTS: 37/422 preterm infants (8.8%) had mucocutaneous colonization with candida, none of our patients developed systemic candidiasis . In 7 mechanically ventilated patients (1.9%) Candida albicans or Candida tropicalis was repeatedly detected in the bronchial secretions; 1 patient who had invasive Candida-pneumonia was effectively treated with 5-Fluocytosin and Fluconazol . 4/352 (1.1%) central silastic catheters were colonized with Candida albicans; none of these patients required specific treatment . CONCLUSION: The low rate of mucocutaneous Candida-colonization and invasive infection found in our patients may be explained--at least in part--by epidemiological and obstetrical factor as well as by the procedures of the neonatal management.

J Hosp Infect, 1992 Sep, 22(1), 65 - 72
An outbreak of Candida tropicalis peritonitis in patients on intermittent peritoneal dialysis; Yuen KY et al.; An outbreak of Candida tropicalis peritonitis in intermittent peritoneal dialysis patients during a 6-week period is reported from a general hospital . Five patients were involved and there were three deaths . The strains recovered from affected patients were identical to those recovered from the water baths on the basis of biotyping, antimicrobial susceptibility pattern and chromosomal DNA fingerprints . No further cases were identified on subsequent surveillance after the prohibition of wet-warming of peritoneal dialysate in the hospital.

Mikrobiologiia, 1992 Sep-Oct, 61(5), 887 - 92
{Study of asynchronous and synchronous yeast cultures using flow cytofluorometry}; Ivanov VN et al.; Distribution of Saccharomyces cerevisiae, Candida boidinii and Candida tropicalis cells according to DNA content was investigated using laser flow cytofluorometry . Cells distribution curves according to DNA content possessed two maxima in case the sample belonged to the exponential phase of the asynchronous batch culture, or one maximum in case the sample was from the stationary phase of growth . In synchronous cultures variations of cells distribution curves according to DNA content (age structure of the population) were demonstrated and the curves with one maximum and plateau were observed.

Yeast, 1992 Sep, 8(9), 721 - 34
Antibodies directed against a yeast carboxyl-terminal peroxisomal targeting signal specifically recognize peroxisomal proteins from various yeasts; Aitchison JD et al.; The carboxyl-terminal tripeptide Ala-Lys-Ile is essential for targeting Candida tropicalis trifunctional enzyme (hydratase-dehydrogenase-epimerase) to peroxisomes of both Candida albicans and Saccharomyces cerevisiae (Aitchison,J.D., Murray, W.W . and Rachubinski, R . A . (1991).J . Biol . Chem . 266, 23197-23203) . We investigated the possibility that this tripeptide may act as a general peroxisomal targeting signal (PTS) for other proteins in the yeasts C . tropicalis, C . albicans, Yarrowia lipolytica and S . cerevisiae, and in rat liver . Anti-AKI antibodies raised against the carboxyl-terminal 12 amino acids of trifunctional enzyme were used to search for this PTS in proteins of these yeasts and of rat liver . The anti-AKI antibodies reacted exclusively with multiple peroxisomal proteins from the yeasts C . tropicalis, C . albicans and Y . lipolytica . There was a weak reaction of the antibodies with one peroxisomal protein from S . cerevisiae and no reaction with peroxisomal proteins from rat liver . Antibodies directed against a synthetic peptide containing a carboxyl-terminal Ser-Lys-Leu PTS (Gould, S . J., Krisans, S., Keller, G.-A . and Subramani, S . (1990) . J . Cell Biol . 110,27-34) reacted with multiple peroxisomal proteins of rat liver and with peroxisomal proteins of yeast distinct from those identified with anti-AKI antibodies . These results provide evidence that several peroxisomal proteins of different yeasts contain a PTS antigenically similar to that of C . tropicalis trifunctional enzyme and that this signal is absent from peroxisomal proteins from at least one mammalian system, rat liver.

FEMS Microbiol Lett, 1992 Aug 15, 74(2-3), 149 - 56
Disruption of the gene encoding the secreted acid protease (ACP) in the yeast Candida tropicalis; Sanglard D et al.; The gene for the secreted acid protease (ACP), a potential virulence factor of Candida species, was inactivated in Candida tropicalis by gene disruption . The disruption was performed by cotransformation of an ade2 C . tropicalis mutant with a linear DNA fragment carrying a deletion in ACP, and the replicative vector pMK16 which carries a selectable ADE2 gene marker . Few of the transformants exhibited lower protease secretion levels and were shown to have one deleted and one unaffected ACP copy, since C . tropicalis is a diploid yeast . These transformants were rendered homozygotic for this deletion by mild UV-treatment . One of the homozygotic acp deletion mutants obtained was completely devoid of extracellular protease activity and grew poorly on bovine serum albumin-containing medium . This mutant could be complemented by an ACP fragment inserted in pMK16, but also by an acid protease gene isolated from C . parapsilosis.

Jpn J Antibiot, 1992 Aug, 45(8), 1060 - 4
{Clinical evaluation of flucytosine in patients with urinary fungal infections}; Tokunaga S et al.; The clinical effectiveness of flucytosine (5-FC) was evaluated in 52 patients with urinary fungal infections which were diagnosed because of the presence of both funguria (greater than or equal to 10(4) cfu/ml) and pyuria (greater than or equal to 5 WBC/hpf) . The patients received oral daily doses of 5-FC ranging 20 to 150 mg/kg, with a mean treatment duration of 11 days . The 57 fungus strains isolated from urine specimens consisted of 51 Candida spp . including 28 Candida albicans, 10 Candida glabrata, 4 Candida tropicalis and 6 Trichosporon beigelii . Forty-seven (92.2%) and 4 (66.7%) of the 51 Candida spp . and 6 T . beigelii strains isolated were eradicated by the treatment, respectively, giving an overall eradication rate of 89.5% . Clinical responses in 32 patients, from whom only fungi were submitted, were excellent in 5, moderate in 22, and poor in 5, thus the overall efficacy rate was 84.4%, based on the criteria of Japanese UTI Committee . Of the 18 strains of Candida spp . tested, 5-FC showed 0.20 micrograms/ml or less of the minimum inhibitory concentrations (MICs) against 16 strains and higher than 100 micrograms/ml against the remaining 2 strains . The MICs of the 5 T . beigelii strains tested were between 3.12 and 6.25 micrograms/ml . As adverse reactions, only 2 patients (3.8%) experienced diarrhea but did not require treatment cessation . These results suggest that flucytosine is an effective drug for urinary fungal infections.

Arzneimittelforschung, 1992 Aug, 42(8), 1045 - 8
Synthesis of some hydroxamic acid derivatives of benzimidazole and their antibacterial and antifungal activities; Gunes HS et al.; A number of 1H-benzimidazole-2-propanoic acid derivatives have been synthesized by Phillips method, and their antibacterial and antifungal activities have been tested . The chemical structures of the synthesized compounds were elucidated by spectroscopic (IR, NMR, mass) and elementary analysis . Investigation of antimicrobial activity of the compounds was done by agar dilution technique using bacteria (Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Mycobacterium smegmatis ATCC 607) and yeast-like fungi (Candida albicans, Candida tropicalis, Candida pseudotropicalis, Candida kruzei, Candida globrata) . Among the compounds tested N-hydroxy-3-(1H-benzimidazol-2-yl)-propionamide (Compound 6) showed considerable activity against both Candida albicans and Candida tropicalis.

Kansenshogaku Zasshi, 1992 Aug, 66(8), 1030 - 6
{Limulus test (factor G) and polysaccharides from fungus}; Miyazaki T et al.; Picogram quantifies per ml of endotoxin showed a positive limulus test . The concentration more than ten pg/ml of curdlan ((1-3)-beta-D-glucan) was also positive to a conventional limulus test, which contains a factor G . One nanogram per ml of CM-curdlan was positive, but higher concentration of 10 pg/ml of CM-curdlan decreased its optical density of the conventional limulus test . More than one hundred ng/ml of mannan from Saccharomyces cerevisiae did not react with factor G . However, a culture supernatant of Saccharomyces cerevisiae activated factor G . This result suggested that beta-glucan activated factor G . In addition, culture supernatant of Aspergillus fumigatus, Rhodotorula rubra, Candida parapsilosis, Candida tropicalis, Candida krusei also activated factor G, but Cryptococcus neoformans did not activate it.

Biotechnology (N Y), 1992 Aug, 10(8), 894 - 8
Metabolic engineering of Candida tropicalis for the production of long-chain dicarboxylic acids; Picataggio S et al.; We have engineered an industrial strain of the yeast, Candida tropicalis, for the efficient production of long-chain dicarboxylic acids, which are important raw materials for the chemical industry . By sequential disruption of the four genes encoding both isozymes of the acyl-CoA oxidase which catalyzes the first reaction in the beta-oxidation pathway, alkane and fatty acid substrates have been successfully redirected to the omega-oxidation pathway . Consequently, the conversion efficiency and chemical selectivity of their terminal oxidation to the corresponding dicarboxylic acids has been improved to 100 percent . The specific productivity of the bioconversion has been increased further by amplification of the cytochrome P450 monooxygenase and NADPH-cytochrome reductase genes encoding the rate-limiting omega-hydroxylase in the omega-oxidation pathway . The amplified strains demonstrated increased omega-hydroxylase activity and a 30% increase in productivity compared to the beta-oxidation-blocked strain in fermentations . The bioconversion is effective for the selective terminal oxidation of both saturated and unsaturated linear aliphatic substrates with chain-lengths ranging from 12 carbons to 22 carbons and also avoids the undesirable chain modifications associated with passage through the beta-oxidation pathway, such as unsaturation, hydroxylation, or chain shortening . It is now possible to efficiently produce a wide range of previously unavailable saturated and unsaturated dicarboxylic acids with a high degree of purity.

Mycopathologia, 1992 Jun, 118(3), 139 - 45
Factors involved in the adherence of Candida albicans and Candida tropicalis to protein-adsorbed surfaces . An in vitro study using immobilized protein; Nikawa H et al.; The adherence of Candida albicans and C . tropicalis to protein-adsorbed surfaces was investigated with surface-modified glass slides to which serum or salivary proteins were covalently bound . A specific adherence like a ligand-receptor interaction was observed between C . albicans and mucin- or salivary protein-immobilized glass slides . This interaction was eliminated by deglycosylation of the slides, suggesting that the receptor may be an oligosaccharide(s) contained mucin or saliva . A similar specific interaction was also observed between C . tropicalis and fibrinogen-immobilized glass surfaces . When the numbers of adherent cells to deglycosylated protein-immobilized glass glides were plotted against zeta potentials and contact angles of these protein-immobilized glass slides, a significant correlation was observed between the numbers of adherent cells and zeta potentials in the case of C . albicans (r = -0.87), whereas a significant correlation was observed between cell numbers and contact angles (r = 0.82) in the case of C . tropicalis . These results suggest that the forces governing the adherence of fungi to pellicle in dentures may vary depending upon the surface properties of fungi and substrate.

J Biochem (Tokyo), 1992 Jun, 111(6), 783 - 7
Beta-oxidation of butyrate, the short-chain-length fatty acid, occurs in peroxisomes in the yeast Candida tropicalis; Kurihara T et al.; When an n-alkane-utilizable yeast, Candida tropicalis pK233, was cultivated on butyrate, the fatty acid of shortest chain-length for beta-oxidation, as the sole source of carbon and energy, catalase and the enzymes of the fatty acid beta-oxidation system were inducibly synthesized at high levels . As in the alkane-grown cells, the proliferation of peroxisomes was harmonized with the induction of peroxisomal enzymes . The results of subcellular fractionation and immunoelectronmicroscopy indicated the localization of these enzymes in peroxisomes, not in mitochondria . It was suggested that only peroxisomes have a role in fatty acid beta-oxidation in the yeast cells, unlike in mammalian cells, in which cooperation between peroxisomes and mitochondria is essential.

Cell Struct Funct, 1992 Jun, 17(3), 203 - 7
Immunoelectron microscopic localization of thiolases, beta-oxidation enzymes of an n-alkane-utilizable yeast, Candida tropicalis; Kamasawa N et al.; The location of acetoacetyl-CoA thiolase (T-I) and 3-ketoacyl-CoA thiolase (T-III), enzymes of the fatty acid beta-oxidation system, was studied in n-alkane-grown Candida tropicalis cells by immunoelectron microscopy using a post-embedding method with colloidal gold conjugated IgG . The deposition of gold particles for T-I was detected in the microbodies and cytoplasm and that of gold particles for T-III specifically in the microbodies . The double labeling technique confirmed that T-I and T-III occurred concurrently in a microbody and T-I also in cytoplasm . These results were consistent with the biochemical data based on subcellular fractionation and indicated that the yeast beta-oxidation system operates efficiently only in the microbodies.

Infection, 1992 May-Jun, 20(3), 132 - 5
Surveillance of nosocomial fungal infections in a burn care unit; Chakrabarti A et al.; A survey was conducted to trace the source of nosocomial fungal infections in the burn care unit of Nehru Hospital, Chandigarh, India, by collection of samples from wounds of 25 severely burnt patients and their surroundings . The environmental sampling revealed predominant fungal contamination by dematiceous hyphomycetes, aspergilli, Penicillium, Fusarium and yeasts (Candida albicans, Candida tropicalis, Candida krusei, Candida parapsilosis), whereas the colonising or invading fungi from the patients were Aspergillus flavus and yeasts of the genus Candida (C . albicans, C . tropicalis, C . krusei, C . parapsilosis, Torulopsis glabrata) . This study thus corroborates the more pathogenic potential of some of the environmental fungal isolates located in the vicinity of the immunocompromised patients and stresses the need for decontamination of the environment of the burn care unit.

Kansenshogaku Zasshi, 1992 May, 66(5), 612 - 9
{Analysis of 84 cases with fungemia}; Nakazawa S et al.; Eighty-four patients with fungemia were analyzed . Fungi had been isolated by culture of blood samples, including blood from the catheter for intravenous hyperalimentation, between 1986-1990 . Candida albicans (39.3%), Candida parapsilosis (20.2%), Candida tropicalis (11.9%), Candida glabrata (10.7%), Candida guilliermondii (4.8%) and Trichosporon beigelii (4.8%) were the most frequently isolated fungal pathogens . Four patients' blood yielded two different fungal species . Fifty-nine cases were male, and 25 cases were female . Forty-six of the 84 patients died (54.8%), but there were no differences in the overall mortality rate as a function of the fungal species or sex . All patients had underlying diseases: solid cancer, 37 cases; cardiovascular diseases, 9 cases; gastrointestinal diseases excluding gastrointestinal cancer, 8 cases; central nervous system diseases, 7 cases; premature infants and congenital abnormality, 7 cases; leukemia, 6 cases and miscellaneous, 10 cases . Twenty-four of the 46 dead cases were autopsied, and eight cases showed systemic fungal lesions . However, in one case of pulmonary cryptococcosis and one case of pulmonary penicilliosis, there was no correlation between the isolation of C . glabrata by blood culture and the pathological findings . A fungus-positive blood culture was surmised to be a result of contamination of the sample in 33 cases, and the mortality rate for those cases was 72.2% (24 cases) . For 6 of the corpses, fungal lesions observed at autopsy were compatible with the types of lesions found by the fungi which had been isolated before death . Removal of the catheter reduced the mortality rate to 41.7% . Fungal endophthalmitis was diagnosed in six cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur J Epidemiol, 1992 May, 8(3), 444 - 51
DNA relatedness, karyotyping and gene probing of Candida tropicalis, Candida albicans and its synonyms Candida stellatoidea and Candida claussenii; Mahrous M et al.; Isolates of Candida albicans with varied phenotypes, including sucrose-negative variants (C . stellatoidea, serotypes A and B) and avirulent germ tube-negative forms (C . claussenii) showed significant (greater than 90%) DNA relatedness to classical C . albicans, but insignificant relatedness to C . tropicalis and sucrose-negative C . tropicalis . A transverse alternating-field gel electrophoresis procedure (TAFE) showed discrete karyotype patterns among the phenotypic variants of C . albicans including the sucrose-negative C . stellatoidea . The number of chromosome-sized DNA bands for C . tropicalis (7 bands) were within the range of bands observed for C . albicans (5 to 10 bands) . The general DNA-migration pattern for C . albicans appeared distinct from that of C . tropicalis . An aspartyl proteinase (PrA) gene probe from C . albicans hybridized with chromosomal DNA from C . albicans, C . claussenii and C . stellatoidea but not with that from C . tropicalis.

Clin Infect Dis, 1992 Apr, 14(4), 952 - 4
Treatment of candidemia in critically ill surgical patients with intravenous fluconazole; Nolla-Salas J et al.; Six nonneutropenic, critically ill patients with candidemia who underwent surgery were treated with intravenous fluconazole, a new, nontoxic triazole derivative . The portal of entry for Candida species could be demonstrated for four patients (the peritoneal cavity in two and a central venous catheter in two) . There were three cases of fungemia due to Candida albicans; two cases were due to Candida tropicalis, and one case was due to Candida parapsilosis . Fluconazole was administered to these patients for a mean of 20 days at doses ranging from 100 to 200 mg/d . All patients survived and Candida species were eradicated from all sites . The decision to treat patients with proven or suspected systemic candidiasis has been made easier by the development of new, nontoxic antifungal agents.

Clin Infect Dis, 1992 Apr, 14(4), 875 - 83
Vascular catheter-associated fungemia in patients with cancer: analysis of 155 episodes; Lecciones JA et al.; We reviewed all 155 episodes of central venous catheter-associated fungemia among inpatients at the National Cancer Institute during a 10-year period . Candida species accounted for 98% of episodes . Fungemia was documented by culture of blood drawn through catheters in 50% of cases and by culture of both catheter-drawn and peripheral blood in 39%; mortality and the rate of dissemination were similar for these two groups . Four management strategies were used: catheter removal, antifungal therapy (with amphotericin B), both, or neither; indications for the use of both modes of treatment included fever, neutropenia, long-term indwelling catheterization, positive cultures of both catheter-drawn and peripheral blood, isolation of Candida tropicalis, and fungal isolation from two or more blood cultures . Disseminated fungal infection was documented in 82% of cases with these features but also in 35% of the less severe cases treated only with catheter removal . In addition, nine (82%) of 11 cases managed only with antifungal therapy had a negative outcome (either death from disseminated infection or the recurrence of fevers and/or fungemia), a finding suggesting that intravascular catheters should be removed in fungemia . Virtually all cases of catheter-associated fungemia in patients with cancer are clinically significant and require prompt therapy with amphotericin B.

Clin Infect Dis, 1992 Mar, 14(3), 756 - 66
Candida parapsilosis: epidemiology, pathogenicity, clinical manifestations, and antimicrobial susceptibility; Weems JJ Jr; Early reports associated Candida parapsilosis with endocarditis in intravenous narcotic addicts . More recently, this species has emerged as an important nosocomial pathogen, with clinical manifestations including fungemia, endocarditis, endophthalmitis, septic arthritis, and peritonitis, all of which usually occur in association with invasive procedures or prosthetic devices . Outbreaks of C . parapsilosis infections have been caused by contamination of hyperalimentation solutions, intravascular pressure monitoring devices, and ophthalmic irrigating solution . Experimental studies have generally shown that C . parapsilosis is less virulent than Candida albicans or Candida tropicalis . However, characteristics of C . parapsilosis that may relate to its increasing occurrence in nosocomial settings include frequent colonization of the skin, particularly the subungual space, and an ability to proliferate in glucose-containing solutions, with a resultant increase in adherence to synthetic materials . Recently developed molecular techniques may facilitate the continued exploration of the epidemiology and pathogenesis of C . parapsilosis infections.

Diagn Microbiol Infect Dis, 1992 Mar-Apr, 15(3), 233 - 7
Yeast in blood cultures . Evaluation of factors influencing outcome; Bryce EA et al.; Sixty episodes of candidemia occurring between January 1987 and September 1989 were retrospectively reviewed and the outcome was compared for three patient categories: neutropenic, postsurgical, and nonneutropenic nonsurgical . Because of increasing reports of resistant yeasts, minimal inhibitory concentrations (MICs) to amphotericin B (amB) were determined to examine its relationship to patient outcome . No significant differences between patient categories were found for selected risk factors, mortality, or species isolated, although there was a tendency for Candida tropicalis to occur in neutropenic patients . Statistical analysis using logistic regression showed that both an amB dose greater than 500 mg and a yeast MIC greater than 0.78 micrograms/ml were independent prognosticators of patient outcome.

Clin Infect Dis, 1992 Mar, 14 Suppl 1, S120 - 5
Candidemia in immunocompromised patients; Meunier F et al.; This study reviews data on patients with fungemia and confirms the high prevalence (50%) of infections caused by non-albicans species of Candida . Fungemia due to C . albicans or Torulopsis glabrata occurred significantly more often in patients with a solid tumor, while fungemia due to Candida tropicalis or Candida krusei was significantly more common in patients with hematologic malignancy (P = .001) . For 31% of patients, only a single blood culture was positive for yeasts, and the prognosis for these patients was not significantly different than that for patients with three or more positive blood cultures (P = 1), including those who had C . albicans fungemia . The overall mortality rate was 41.8%, which is much lower than that previously reported in studies of patients with cancer . No significant difference was observed between patients treated with amphotericin B and those treated with fluconazole in this retrospective analysis . Although no significant difference was observed in the mortality rate among patients who had fungemia with or without neutropenia, the incidence of disseminated candidiasis was significantly higher among neutropenic patients (P = .03).

Biochem J, 1992 Mar 1, 282 ( Pt 2), 325 - 31
Purification and some properties of alcohol oxidase from alkane-grown Candida tropicalis; Dickinson FM et al.; Alcohol oxidase was purified to homogeneity from membrane fractions obtained from alkane-grown Candida tropicalis . The enzyme appears to be a dimer of equal-sized subunits of Mr 70000 . The purified enzyme is photosensitive and contains flavin-type material which is released by a combination of boiling and proteolytic digestion . The identity of the flavin material is not yet known, but it is not FMN, FAD or riboflavin . The enzyme is most active with dodecan-I-ol, but other long-chain alcohols are also attacked . The enzyme shows a weak, but significant activity towards long-chain aldehydes . Detailed kinetic studies with decan-1-ol as substrate suggest a group-transfer (Ping-Pong)-type mechanism of catalysis.

J Pediatr, 1992 Mar, 120(3), 455 - 61
Outbreak of Candida bloodstream infections associated with retrograde medication administration in a neonatal intensive care unit; Sherertz RJ et al.; An outbreak of candidemia involving five infants receiving total parenteral nutrition in the neonatal intensive care unit was investigated . Cultures of the intravenous fluids demonstrated that the retrograde medication syringe fluids were significantly more likely to be contaminated with Candida than were other fluids being administered to the infants (p less than 0.001) . Candidemia was significantly associated with total parenteral nutrition (p = 0.04) and retrograde medication administration (p = 0.02) . A survey of nursing practice found that reuse of the retrograde syringes was the most likely cause of contamination . Molecular typing showed that the strains of Candida albicans that were isolated from the bloodstream were also found in the retrograde syringes and that at least three strains of C . albicans and one strain each of Candida tropicalis and Candida parapsilosis were involved . In vitro growth curves demonstrated that Candida species had a selective growth advantage versus bacteria in the total parenteral nutrition fluid . An in vitro simulation of the retrograde medication administration system suggested that the outbreak probably developed after the frequency of changing intravenous tubing was decreased from every 24 hours to every 72 hours . The outbreak was terminated by using syringes only once and resuming intravenous tubing changes every 24 hours . Retrograde medication administration in association with total parenteral nutrition may increase the risk of Candida line infection.

Farmaco, 1992 Feb, 47(2), 219 - 28
Antimicrobial and genotoxic activities of N-substituted 2,2'-dicarboxamidodiphenyldisulfides; Zani F; This paper describes the in vitro evaluation of antibacterial, antifungal and genotoxic activities of a series of 2,2'-dicarboxamidodiphenyldisulfides . All the studied compounds exhibited antibacterial activity against Bacillus subtilis . Most compounds were also active against Staphylococcus aureus . Several compounds were active against Saccharomyces cerevisiae and, in most cases, also against Candida tropicalis . No effects were observed against Escherichia coli or Aspergillus niger . Antimicrobial activity turned out to be affected by the substituent in the benzene ring and structure-activity relationships were found . The antibacterial and antifungal activities of the studied derivatives were compared with those of the corresponding 1,2-benzisothiazolin-3-ones and in some cases strong differences were observed . None of the tested compounds contained alerting groups or showed genotoxic properties.

Farmaco, 1992 Feb, 47(2), 203 - 17
Synthesis and evaluation of antimicrobial activity of new 4-nitroso and 4-diazopyrazole derivatives; Daidone G et al.; Some N-(pyrazol-5-yl)-2-nitrobenzamides, variably substituted in the pyrazole nucleus as well as in the amidic group, were reacted in acetic acid media with potassium nitrite and hydrochloric acid . The different chemical behaviour of the reacted pyrazole derivatives in relation to the substitution pattern in both the pyrazole nucleus and the amidic group, was observed . All compounds isolated from the reaction mixtures (4-nitroso, 4-nitro, 4-diazo and 4-chloro derivatives) were evaluated by the agar diffusion method for their "in vitro" growth inhibitory activity against Candida albicans (our collection), Candida tropicalis ATCC 13803, Saccharomyces cerevisiae ATCC 36375, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853 . The 1-methylpyrazole derivatives showed larger inhibition zones than the 1-phenyl ones in the antimicrobial tests.

Eur J Clin Microbiol Infect Dis, 1992 Feb, 11(2), 152 - 6
Effects of fluconazole on the sterol and carbohydrate composition of four species of Candida; Pfaller M et al.; The effects of fluconazole, a bis-triazole antifungal agent, on the sterol and carbohydrate composition of Candida albicans, Candida tropicalis, Candida krusei and Candida parapsilosis were investigated . Exposure of Candida species to fluconazole resulted in a profound depletion of ergosterol with a corresponding increase in lanosterol content versus control cells . Carbohydrate analysis revealed a significant increase in chitin and either a decrease (Candida albicans, Candida tropicalis and Candida parapsilosis) or an increase (Candida krusei) in glucan content in fluconazole-treated cells . The decreased ergosterol and increased lanosterol content is consistent with 14-alpha-demethylase inhibition by fluconazole . The increase in cell wall chitin is most likely due to deregulation of chitin synthesis secondary to ergosterol depletion in the cell membrane . Because chitin, glucan and ergosterol are critical components of the fungal cell, perturbation of the production and localization of these components by fluconazole is likely to contribute to the selective toxicity of this compound to Candida species and other fungi.

Appl Microbiol Biotechnol, 1992 Feb, 36(5), 650 - 4
Targeted integrative transformation of Candida tropicalis by electroporation; Rohrer TL et al.; A method for integrative transformation of the diploid yeast Candida tropicalis by electroporation has been developed . by linearizing the transforming plasmid DNA containing the URA3 gene prior to electroporation of recipient cells, its integration was targeted to a specific locus in the genome, resulting in single or multiple tandem integrations . The optimal electroporation conditions for this yeast were established and include an electric pulse of 2.25 kV/cm for a duration of 50 ms . Using these conditions, Ura+ transformants were readily obtained at a high frequency (45 transformants/micrograms DNA) as the result of targeted integration of the URA3 gene containing plasmid DNA at the chromosomal ura3 locus . The number of transformants resulting from this procedure is comparable to that achieved with a recently reported spheroplast transformation procedure for C . tropicalis; in addition, it offers the advantages of being simple, rapid and reproducible.

J Med Chem, 1992 Jan, 35(1), 100 - 6
Synthesis, specificity, and antifungal activity of inhibitors of the Candida albicans delta 24-sterol methyltransferase; Ator MA et al.; A series of side chain modified analogues of cholesterol and lanosterol (1-10) have been synthesized and evaluated as inhibitors of the Candida albicans delta 24-sterol methyltransferase . Two sterol substrate analogues 1 and 2 which contained a 24-thia substituent were relatively modest inhibitors of the enzyme (Ki = 1.5-72 microM) . Compounds which mimic the carbocation intermediates proposed for the methyltransferase reaction, including sulfonium salts 4-6, amidines 7 and 8, and imidazoles 9 and 10 were substantially more potent inhibitors (Ki = 5-500 nM) . All of the sterol analogues examined displayed less than 10-fold selectivity for inhibition of the methyltransferase versus the rat liver delta 24-sterol reductase . The sterol analogues were tested for in vitro antifungal activity against C . albicans, Candida tropicalis, and Torulopsis glabrata . The minimum inhibitory concentrations versus C . albicans correlated well with the Ki values for methyltransferase inhibition, and the potency of several compounds approached that of amphotericin B, although only modest fungicidal activity was observed.

Biochem Cell Biol, 1992 Jan, 70(1), 76 - 80
Fatty acid uptake in Candida tropicalis: induction of a saturable process; Trigatti BL et al.; The rates of oleate uptake by Candida tropicalis cells grown on a high oleate concentration (3.5 mM oleate in the presence of 0.50% Brij 58) were higher than those observed in cells grown on glucose; however, oleate uptake was not saturable with substrate concentration . Cells grown at a low oleate concentration (1.0 mM oleate and 2.5% Brij 58) grew to a lower density and at a slightly slower rate; these cells were found to take up oleate at a rate 43-fold higher than cells grown on high oleate concentration . Furthermore, oleate uptake by the cells grown in low oleate medium was a saturable process with Kt and Vmax values of 56 microM and 15 nmol/(min.mg cell protein), respectively . The growth of C . tropicalis under low fatty acid concentration thus clearly results in the induction of a saturable process for its uptake . The total level of acyl-CoA synthetase activity in cells grown on the low oleate concentrations was only twofold higher than in high oleate or glucose grown cells; the level of this enzyme thus does not account for the saturable process and suggests that either the enzyme is regulated in vivo or else a hitherto unidentified enzyme is induced by growth in low concentrations of oleate.

Microbiol Immunol, 1992, 36(6), 637 - 41
Purification and characterization of secretory proteinase of Candida albicans; Yamamoto T et al.; Proteolytic activity of medically important yeasts was tested in both YCB-BSA agar and medium . All of 134 strains of Candida albicans, 13 of 18 strains of Candida tropicalis and 11 of 18 strains of Candida parapsilosis had this activity, while none of 52 Candida glabrata strains or of 11 Cryptococcus neoformans strains tested had proteolytic activity . Strains of C . albicans fell into five groups based on the level and time-course of in vitro proteinase productivity . Five strains randomly selected from each group were tested for pathogenicity in mice . The strain possessing the strongest pathogenicity was used to purify proteinase . The molecular weight of the proteinase was approximately 44,000 daltons and its isoelectric point was pH 4.2 . Optimal pH of the proteinase was 3.2 and the enzyme was stable below pH 7.0 and lost its activity above pH 8.0 at 37 C in a 60-min incubation . The 23 amino acid sequence of the proteinase N-terminus was determined.

Microbiol Immunol, 1992, 36(10), 1099 - 104
Comparison of secretory acid proteinases from Candida tropicalis, C . parapsilosis and C . albicans; Sono E et al.; Acid proteinases secreted by Candida tropicalis and C . parapsilosis were newly isolated . Their physico-chemical and enzymatic properties of molecular weight, pH stability, isoelectric points, specific activity, and N-terminal amino acid sequences were determined and compared with those of a C . albicans acid proteinase . The two acid proteinases secreted by C . parapsilosis were found to be new enzymes in their molecular weights . The acid proteinases from C . tropicalis and C . parapsilosis showed lower activity at neutral pH, less resistance to neutral and alkaline pH than that from C . albicans, and a half or a third of the specific activity of the C . albicans enzyme . These differences seemed to be associated with the difference of pathogenesis between Candida species . Of the 31 N-terminal amino acids, the enzymes of these three Candida species revealed 12 homologous amino acids.

Chemotherapy, 1992, 38(5), 335 - 9
In vitro susceptibility of 245 yeast isolates to amphotericin B, 5-fluorocytosine, ketoconazole, fluconazole and itraconazole; Martin E et al.; The in vitro activity of amphotericin B, 5-fluorocytosine, ketoconazole, fluconazole and itraconazole was tested against 245 yeast strains isolated from clinical specimens (68 Candida albicans, 74 Candida tropicalis, 43 Candida krusei, 28 Candida glabrata, 19 Candida parapsilosis, 8 Candida lusitaniae and 5 Candida guilliermondii) . An agar dilution method was employed to carry out testing . Minimal inhibitory concentrations to restrain 90% of isolate growth (MIC90) ranged from 0.12 to 2 mg/l for amphotericin B and for 5-fluorocytosine, from 0.03 to 8 mg/l for ketoconazole, from 0.05 to 50 mg/l for itraconazole and from 0.1 to > 100 mg/l for fluconazole . Among the azole derivatives, the most active was ketoconazole, followed by itraconazole . Only 1 strain of C . albicans was resistant to amphotericin B (MIC > 4 mg/l) . Both C . tropicalis and C . krusei responded poorly to fluconazole and the former to itraconazole as well . The species most susceptible to the antifungal agents tested was C . glabrata and the most resistant were C . tropicalis and C . krusei.

Nauchnye Doki Vyss Shkoly Biol Nauki, 1992, (4), 90 - 100
{The effect of Bacillus intermedius RNAse on the multiplication of Candida tropicalis yeasts}; Kupriianova-Ashina FG et al.; The effect of Bacillus intermedius RNAse on the reproduction of Candida tropicalis and synthesis of the main biopolymers in the yeast cells . It has been found that stimulating action of the enzyme appears at the concentration of 10(-5)-10(-6) mg/ml and does not depend on the physiological state of the sowing culture . The connection between the increase of the ionic penetration and stimulation of the RNA and proteins synthesis in the yeast cells subjected to the RNAse action is shown . The mechanism of chromatine-associated RNA-polymerase activation is suggested to include the alteration of the ionic penetration of cells under the RNAse action.

J Biol Chem, 1991 Dec 5, 266(34), 23197 - 203
The carboxyl-terminal tripeptide Ala-Lys-Ile is essential for targeting Candida tropicalis trifunctional enzyme to yeast peroxisomes; Aitchison JD et al.; The gene encoding Candida tropicalis peroxisomal trifunctional enzyme, hydratase-dehydrogenase-epimerase (HDE), was expressed in both Candida albicans and Saccharomyces cerevisiae . The cellular location of HDE was determined by subcellular fractionation followed by Western blot analysis of peroxisomal and cytosolic fractions using antiserum specific for HDE . HDE was found to be exclusively targeted to and imported into peroxisomes in both heterologous expression systems . Deletion and mutational analyses were used to determine the regions within HDE which are essential for its targeting to peroxisomes . Deletion of a carboxyl-terminal tripeptide Ala-Lys-Ile completely abolished targeting of HDE to peroxisomes, whereas large internal deletions of HDE (amino acids 38-353 or 395-731) had no effect on HDE targeting to peroxisomes in either yeast . This tripeptide is similar to, but distinct from, other tripeptide peroxisomal targeting sequences (PTSs) as identified in peroxisomal firefly luciferase and four mammalian peroxisomal proteins . Substitutions within the carboxyl-terminal tripeptide (Ala----Gly and Lys----Gln) supported targeting of HDE to peroxisomes of C . albicans but not of S . cerevisiae . This is the first detailed analysis of the peroxisomal targeting signal in a yeast peroxisomal protein.

J Dermatol, 1991 Dec, 18(12), 695 - 706
H1 histones contribute to candidacidal activities of human epidermal extract; Kashima M; In the study of the purification of candidacidal cationic proteins from human epidermis by using high performance liquid chromatography, it was found that these proteins were composed of several groups . Among them, the most active group was purified . The amino acid compositions of this protein group were almost the same as those of human splenic H1 histones . The results of immunoblotting suggested that the proteins were human epidermal H1 histones . The killing speed of the epidermal H1 histone was very rapid, 8 micrograms of the protein killed 90% of Candida tropicalis (1 x 10(5) CFU) within 10 minutes . The cidal activity increased in lower pH conditions and decreased at higher ionic strengths . Because nuclei of the epidermis disintegrate in the granular layer, it is suggested that, in this layer, nuclear histones may be released from the nuclei . My immunohistochemical results suggest that H1 histones may contribute to form a barrier which inhibits candida from invading deeper than the granular cell layer in cases of skin candidial infection.

J Biochem (Tokyo), 1991 Dec, 110(6), 909 - 14
Presence of two transcribed malate synthase genes in an n-alkane-utilizing yeast, Candida tropicalis; Hikida M et al.; The presence of two genomic DNA regions encoding malate synthase (MS) was shown by Southern blot analysis of the genomic DNA from an n-alkane-assimilating yeast, Candida tropicalis, using a partial MS cDNA probe, in accordance with the fact that two types of partial MS cDNAs have previously been isolated . This was also confirmed by the restriction mapping of the two genes screened from the yeast lambda EMBL library . Nucleotide sequence analysis of the respective genomic DNAs, named MS-1 gene and MS-2 gene, revealed that both regions encoding MS had the same length of 1,653 base pairs, corresponding to 551 amino acids (molecular mass of MS-1, 62,448 Da; MS-2, 62,421 Da) . Although 29 nucleotide pairs differed in the sequences of the coding regions, the number of amino acid replacements was only one: 159Asn (MS-1)----159Ser (MS-2) . In the 5'-flanking regions, there were replacements of four nucleotide pairs, deletion of one pair, and insertion of four pairs . In spite of the fact that two genomic genes were present and transcribed, RNA blot analysis demonstrated that only one band (about 2 kb) was observable even when the carbon sources in the cultivation medium were changed . A comparison of the amino acid sequences was made with MSs of rape (Brassica napus L.), cucumber seed, pumpkin seed, Escherichia coli, and Hansenula polymorpha . A high homology was observed among these enzymes, the results indicating that the protein structure was relatively well conserved through the evolution of the molecule.(ABSTRACT TRUNCATED AT 250 WORDS)

J Infect Dis, 1991 Nov, 164(5), 928 - 35
Candida albicans stimulates endothelial cell eicosanoid production; Filler SG et al.; The response of human umbilical vein endothelial cells to invasion by Candida species was examined in vitro . Live Candida albicans caused significant endothelial release of eicosanoids, mainly prostaglandins . Since prostaglandin I2 (PGI2) is an important prostaglandin produced by endothelial cells, factors influencing its release were studied . The ability of different strains and species of Candida to induce endothelial PGI2 release was closely related to their capacity to injure the endothelium (r = .99) . C . albicans was the only species tested that either stimulated PGI2 release or damaged the endothelial cells; only this organism possessed detectable phospholipase activity . Candida glabrata and Candida tropicalis had no phospholipase activity and neither increased PGI2 release nor caused significant endothelial damage . Close proximity with germinated C . albicans was required for endothelial injury and PGI2 release . The ability of C . albicans to stimulate endothelial cells may have important implications in regulating neutrophil activities against organisms that interact with endothelial cells.

Zentralbl Bakteriol, 1991 Oct, 275(4), 504 - 12
Mode of action of the antimycotic agent G2 isolated from alfalfa roots; Polacheck I et al.; The mode of action of the antimycotic alfalfa root saponin, medicagenic acid 3-O-beta-D-glucopyranoside (compound G2), which possesses a pronounced antifungal activity against medically important yeasts and dermatophytes, was studied in Saccharomyces cerevisiae . Compound G2 caused lethal leakage of ions out of the yeast cells . Exposure of S . cerevisiae to compound G2 resulted in a disappearance of the main sterol, ergosterol, from the cell membranes, suggesting that compound G2 was highly specific for ergosterol . Independently, chemical data indicated that compound G2 forms stable complexes with both ergosterol and cholesterol . Addition of cholesterol or ergosterol protected the cells of S . cerevisiae and several pathogenic yeasts from the inhibitory activity of compound G2 by producing a higher ratio of sterols (mainly ergosterol) to phospholipids in the membranes . The fact that an amphotericin B-resistant Candida tropicalis was susceptible to G2 suggested that its mode of action was different from that described for polyene antibiotics . This was also confirmed by the finding that 0.2 M KCl did not protect S . cerevisiae cells against ion leakage with G2, but did so with amphotericin B.

Appl Microbiol Biotechnol, 1991 Oct, 36(1), 48 - 60
Optimization of Candida tropicalis cytochrome P450alk gene expression in Saccharomyces cerevisiae with continuous cultures; Beretta I et al.; The cytochrome P450alk gene (P45alk) from Candida tropicalis ATCC 750 was expressed in Saccharomyces cerevisiae GRF18 under control of the alcohol dehydrogenase I (ADHI) promoter . To achieve stable expression over long time periods, a 2-microns derived replicative and an integrative expression system were tested in continuous culture . The 2-microns derived replicative system could not be maintained in cells over high generation numbers . In continuous culture, the instability was more pronounced at high dilution rates (D) and high histidine concentration, for which the yeast is auxotrophic . The nature of the instability was probably due to a gene conversion event between the plasmid and the yeast chromosome . In contrast, the integrative expression system was stably maintained in cells over prolonged cultivation times . Since this work focused on the production of large quantities of P450 by heterologous expression in yeast over prolonged time periods, the integrant was used to optimize P450alk expression by varying continuous culture parameters . The P450alk expression was shown to be dependent on the D applied to the culture . The highest P450alk expression levels were obtained at high D, when cell metabolism was shifted to partial glucose oxidation, yielding ethanol as a major metabolite in the culture supernatant . In contrast, when glucose was completely oxidized at low D, the ADHI-dependent P450alk expression was reduced and followed by a corresponding decrease in heterologous protein.

Gene, 1991 Sep 30, 106(1), 61 - 9
Sequences of two tandem genes regulated by carbon sources, one being essential for n-alkane assimilation in Candida maltosa; Hwang CW et al.; Several n-alkane-inducible clones were isolated from the genomic library of an n-alkane-assimilation yeast, Candida maltosa, by the differential hybridization method . Among these, one of the most predominantly expressed clones was analyzed . The nucleotide sequence of the cloned DNA fragment showed that it contained two open reading frames, one encoding a protein of 127 amino acids (aa) and the other a protein of 276 aa . The former was named POX18Cm, because the sequence was highly homologous to that of the Candida tropicalis gene, POX18, which already had been identified as encoding a small oleate-inducible peroxisomal protein . The latter, named ALI1, had no homologous sequences in the EMBL database (1990 release) . Northern-blot hybridization indicated that the expression of these two genes was regulated by carbon sources in the media . From gene-disruption experiments, it was concluded that ALI1 was essential for assimilation of n-alkane by C . maltosa.

Gene, 1991 Sep 30, 106(1), 51 - 60
Characterization of a second alkane-inducible cytochrome P450-encoding gene, CYP52A2, from Candida tropicalis; Seghezzi W et al.; A second alkane-inducible cytochrome P450-encoding gene (CYP52A2) from the yeast Candida tropicalis was sequenced and characterized . CYP52A2 is located 1 kb upstream from CYP52A1, the previously characterized P450 gene {Sanglard and Loper, Gene 76 (1989) 121-136} and shows the same orientation . Like CYP52A1, CYP52A2 is induced by growth on alkane . Both promoter regions share repeats of the sequence CATGTGAA that could be of importance for the induction of the two genes . At the amino acid level, alk2 shows an overall identity of 68.2% and an overall similarity of 81.6% to alk1 . Regions of high homology between the two proteins are found in the distal and proximal heme binding sites which contain the highly conserved cysteine residue as the fifth ligand to the heme iron . However, marked differences between the two proteins exist at their N-terminal end, which includes the transmembrane domain, and at the putative substrate-binding domain . Upon expression of CYP52A2 in Saccharomyces cerevisiae, alk2 was shown to hydroxylate hexadecane, but had no hydroxylation activity towards lauric acid, whereas alk1 showed both activities . Comparative immunoblots demonstrate that neither alk1 nor alk2 expressed in S . cerevisiae corresponds to the main cytochrome P450 present in C . tropicalis when grown on alkane.

Br J Ophthalmol . 1991 Sep;75(9):565.
Candida tropicalis endophthalmitis following penetrating keratoplasty; Behrens-Baumann W et al.; A case of Candida tropicalis endophthalmitis following penetrating keratoplasty is presented . The donor was an alcoholic, who died of bronchopneumonia and pancreatitis . We presume the candida infection was transmitted by the donor because Candida tropicalis was cultured in life from the donor's throat swab and corresponding fungal elements were discovered post mortem in kidney sections of the donor.

Infect Immun, 1991 Sep, 59(9), 2972 - 7
Molecular cloning of the secretory acid proteinase gene from Candida albicans and its use as a species-specific probe; Ganesan K et al.; Candida albicans secretes an acid proteinase when grown with a protein as a sole nitrogen source . The gene encoding this proteinase was isolated from a genomic expression library of C . albicans constructed in lambda gt11 by screening with antiproteinase antibodies . The affinity-purified antibodies used to verify the clones are monspecific; these do not cross-react with any other protein in the culture supernatants or crude extracts of C . albicans but strongly react with fusion proteins encoded by recombinant clones, revealing that these are true proteinase clones . Genomic Southern blot analysis shows that the proteinase gene is present at a unique locus and that there is no other closely related gene in the C . albicans genome . The proteinase gene probe identified two transcripts on Northern blots (RNA blots), which are present at a much higher level in C . albicans cells induced for proteinase secretion than in uninduced cells . The aspartyl proteinase gene reported earlier (T.J . Lott, L.S . Page, P . Boiron, J . Benson, and E . Reiss, Nucleic Acids Res . 17:1779, 1989) is not that of secretory acid proteinase, since the N-terminal amino acid sequence of secretory acid proteinase does not correspond to the deduced amino acid sequence of the aspartyl proteinase gene . The secretory acid proteinase gene was used to probe Southern blots of genomic DNA of several medically important Candida species and Saccharomyces cerevisiae . Under hybridization and wash conditions of low stringency, Candida tropicalis and Candida parapsilosis, in addition to C . albicans strains, gave specific signals, implying that C . tropicalis and C . parapsilosis have homologous secretory acid proteinase genes . However, under under wash conditions of high stringency, signals were obtained only with C . albicans strains, suggesting that this gene can be used as a species-specific probe . A simple yeast colony hybridization technique is sufficient to distinguish C . albicans from other yeasts.

Mol Cell Biol, 1991 Sep, 11(9), 4333 - 9
Determination of Candida tropicalis acyl coenzyme A oxidase isozyme function by sequential gene disruption; Picataggio S et al.; A recently developed transformation system has been used to facilitate the sequential disruption of the Candida tropicalis chromosomal POX4 and POX5 genes, encoding distinct isozymes of the acyl coenzyme A (acyl-CoA) oxidase which catalyzes the first reaction in the beta-oxidation pathway . The URA3-based transformation system was repeatedly regenerated by restoring the uracil requirement to transformed strains, either through selection for spontaneous mutations or by directed deletion within the URA 3 coding sequence, to permit sequential gene disruptions within a single strain of C . tropicalis . These gene disruptions revealed the diploid nature of this alkane- and fatty acid-utilizing yeast by showing that it contains two copies of each gene . A comparison of mutants in which both POX4 or both POX5 genes were disrupted revealed that the two isozymes were differentially regulated and displayed unique substrate profiles and kinetic properties . POX4 was constitutively expressed during growth on glucose and was strongly induced by either dodecane or methyl laurate and to a greater extent than POX5, which was induced primarily by dodecane . The POX4-encoded isozyme demonstrated a broad substrate spectrum in comparison with the narrow-spectrum, long-chain oxidase encoded by POX5 . The absence of detectable acyl-CoA oxidase activity in the strain in which all POX4 and POX5 genes had been disrupted confirmed that all functional acyl-CoA oxidase genes had been inactivated . This strain cannot utilize alkanes or fatty acids for growth, indicating that the beta-oxidation pathway has been functionally blocked.

Hinyokika Kiyo, 1991 Sep, 37(9), 969 - 74
{Candida urinary tract infection with special reference to ascending pyelonephritis}; Ohkawa M et al.; The pathogenesis of Candida urinary tract infection (UTI) has been investigated clinically and experimentally with special reference to ascending pyelonephritis in rats . Among the Candida species, Candida albicans was most frequently isolated from clinical specimens including urine in two medical centers, one in Japan and the other in the United States . The isolates of C . albicans serotype B showed a significantly lower susceptibility to 5-fluorocytosine compared to those of serotype A (p less than 0.01) . The distribution pattern of the serum antibody titers against C . albicans in 20 candiduria patients (C . albicans 19 and Candida tropicalis 1) was similar to that in 23 bacterial complicated UTI patients . All patients with candiduria had underlying diseases of the urinary tract, such as neurogenic bladder, bladder cancer or benign prostatic hyperplasia: indwelling urinary catheters were present in 15 patients and all had received antimicrobial agents before the study . Ascending Candida pyelonephritis has been investigated in female rats which were transurethrally inoculated into the bladder with C . albicans ATCC 10259 strain . The incidence of Candida pyelonephritis was approximately 80% in rats treated with cyclophosphamide and more than 70% in rats with partial ureteral obstruction . There was a significant relationship between renal and urinary Candida cell populations (p less than 0.01) . Furthermore, a significant relationship was revealed between renal Candida cell populations and histological grades of pyelonephritis (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Vnitr Lek, 1991 Sep-Oct, 37(9-10), 800 - 4
{Candida myocarditis and polymyositis in acute lymphoblastic leukemia}; Resl M et al.; Candida sepsis in 18-year-old man with acute lymphoblastic leukemia was associated with multisystem alteration accentuated in cardiac and skeletal muscles . Candida tropicalis identified in the post-mortem blood culture was resistant to azole-chemotherapeutics and semi-sensitive to polyene-antibiotics . Unusual myocardial involvement and a role of Candida spp . during opportune infections in immunosuppressed patients are discussed.

Gene, 1991 Aug 30, 105(1), 135 - 6
Sequence of the gene encoding Candida tropicalis peroxisomal trifunctional enzyme; Aitchison JD et al.; The gene (HDE) encoding the peroxisomal trifunctional enzyme, hydratase-dehydrogenase-epimerase, of the yeast Candida tropicalis, was cloned and its nucleotide sequence is reported.

Gene, 1991 Aug 30, 105(1), 129 - 34
Glucose-responsive and oleic acid-responsive elements in the gene encoding the peroxisomal trifunctional enzyme of Candida tropicalis; Sloots JA et al.; We have investigated the regulation of expression of the gene (HDE), encoding the peroxisomal trifunctional enzyme hydratase-dehydrogenase-epimerase (HDE), of the diploid yeast Candida tropicalis . Heterologous expression in Saccharomyces cerevisiae of constructs containing deletions in the upstream region of the HDE gene has allowed for determination of regions responsible for the control of expression of the HDE gene . Expression was monitored by immunoblot analysis of yeast lysates with anti-HDE serum . Regions have been identified that are responsible for both repression by glucose and induction by oleic acid . A glucose-responsive region lies between nucleotides (nt) -526 and -393 . An oleic acid-responsive region lies between nt -393 and -341 . An additional region controlling derepression by nonfermentable carbon sources is located downstream from nt -341 . Comparison of the nt sequences of these regions to upstream regions of other oleic acid-responsive genes of C . tropicalis has identified possible consensus nt sequences for glucose- and oleic acid-responsive upstream elements in these genes . The regulation of the HDE gene in S . cerevisiae closely resembles that found in C . tropicalis, suggesting that similar mechanisms of transcriptional control operate in both yeasts.

Cancer, 1991 Aug 1, 68(3), 594 - 9
Candida tropicalis and Candida albicans fungemia in children with leukemia; Marina NM et al.; The records were reviewed for all patients hospitalized at a pediatric oncology center for complications of leukemia (n = 822) or lymphoma (n = 290) during an 8-year period . The results of surveillance cultures (throat, rectal, and urine) and blood cultures were analyzed to identify cases of Candida tropicalis and C . albicans colonization and/or fungemia . None of the patients with lymphoma who had positive surveillance cultures for C . albicans (n = 89) or C . tropicalis (n = 23) had fungemia . Among patients with leukemia, significant fungal infection was documented in 12 of 107 colonized with C . tropicalis (11.2%) versus 14 of 700 (2%) colonized with C . albicans (P less than 0.001) . The two groups of children with fungemia were similar in primary diagnoses (predominantly acute lymphoblastic leukemia) and in the frequency of several known risk factors for infection, including the duration of neutropenia (absolute neutrophil counts, less than 500/microliters) . Patients with C . tropicalis fungemia all had disseminated disease compared with nine of 14 patients with C . albicans fungemia . Also, subcutaneous abscesses were unique to patients with C . tropicalis in this series . Two patients in each group died of their infection; central nervous system involvement was present in both fatal cases of C . tropicalis fungemia . A high index of suspicion and the early institution of appropriate antifungal therapy are critical to the successful management of these infections in patients with leukemia.

Postgrad Med J, 1991 Aug, 67(790), 767 - 9
Syringomyelia associated with post-meningitic spinal arachnoiditis due to Candida tropicalis; Phanthumchinda K et al.; A 63 year old man who suffered from syringomyelia related to post-meningitic spinal arachnoiditis caused by Candida tropicalis is reported . The clinical syndrome of syringomyelia developed gradually and a definite diagnosis was delayed for more than 10 years . The patient has partially recovered after surgical treatment . This form of fungal infection and its delayed neurological complication in the form of syringomyelia has not been reported previously, to our knowledge.

Eur J Clin Microbiol Infect Dis, 1991 Aug, 10(8), 665 - 8
Comparison of the in vitro antifungal activity of free and liposome-encapsulated amphotericin B; Anaissie E et al.; One hundred and four pathogenic yeast isolates (32 Candida albicans, 20 Candida tropicalis, 20 Candida parapsilosis and 32 Cryptococcus neoformans) and 21 mould isolates (13 Aspergillus spp . and eight Fusarium spp.), most of which were isolated from patients with cancer, were tested for susceptibility to free amphotericin B and a small unilamellar liposomal formulation of amphotericin B using a microbroth dilution method . Minimal fungicidal concentrations were also determined for Candida spp . and Cryptococcus neoformans . The minimal inhibitory concentrations of liposomal amphotericin B were comparable with those of the free drug; furthermore, the fungicidal activity of these two drugs did not differ significantly.

FEMS Microbiol Lett, 1991 Aug 1, 66(2), 209 - 13
The route of lysine breakdown in Candida tropicalis; Large PJ et al.; Candida tropicalis was found to contain high levels of the following enzymes after growth in defined medium on L-lysine as sole nitrogen source: L-lysine N6-acetyltransferase, N6-acetyl-lysine aminotransferase, and aminotransferase activity for 5-aminovalerate and 4-aminobutyrate . Extracts were also capable of converting 5-acetamidovalerate (and 4-acetamidobutyrate) to acetate . N6-Acetyllysine however, only gave rise to acetate in the presence of 2-oxoglutarate, NAD+ and thiamine pyrophosphate . These activities were undetectable or present in much lower concentrations in cells that had been grown on ammonium sulphate as sole nitrogen source . It is concluded that L-lysine is degraded in this organism via N6-acetyllysine, 5-acetamidovalerate and 5-aminovalerate, both nitrogen atoms being removed by transamination.

FEBS Lett, 1991 Jul 29, 286(1-2), 61 - 3
Genes encoding peroxisomal enzymes are not necessarily assigned on the same chromosome of an n-alkane-utilizable yeast Candida tropicalis; Fukuda Y et al.; We have resolved eight chromosomal bands from an n-alkane-assimilating yeast, Candida tropicalis pK 233, by using contour-clamped homogeneous electric field gel electrophoresis (CHEF) . From the results of hybridization of DNA probes of yeast peroxisomal enzymes--catalase, acyl-CoA oxidase, carnitine acetyltransferase, isocitrate lyase, malate synthase, acetoacetyl-CoA thiolase, and 3-ketoacyl-CoA thiolase--to Southern transfers of CHEF gels, these genes were proven not necessarily to be located on the same chromosome . This fact shows that the genes encoding the enzymes tested were not distributed to be cistronic, although simultaneous and inducible synthesis of peroxisomal enzymes occurred in harmony with the proliferation of peroxisomes, suggesting that their co-ordinated expression might be mainly regulated by certain trans-acting factors.

FEBS Lett, 1991 Jul 29, 286(1-2), 181 - 5
Isolation and nucleotide sequence of the extracellular acid protease gene (ACP) from the yeast Candida tropicalis; Togni G et al.; The extracellular acid protease of Candida tropicalis was purified from the supernatant fraction of culture medium containing bovine serum albumin as nitrogen source and the NH2-terminal amino acid (aa) sequence of the protein was determined . The gene for the acid protease (ACP) was isolated using a pool of synthetic oligonucleotides as a probe and a segment of the deduced aa sequence was found to be in agreement with the NH2-terminal aa sequence of the protein . The deduced aa sequence of ACP is similar to the aa sequence of proteases of the pepsin family . The nucleotide sequence of the 5' portion of this gene revealed a coding sequence for a 60 residue propeptide containing two Lys-Arg amino acid pairs that have been identified as sites for peptidase processing of several exported peptides and proteins . The final Lys-Arg site occurs at the junction with the mature extracellular form of the acid protease.

Kansenshogaku Zasshi, 1991 Jul, 65(7), 838 - 43
{Laboratory and clinical investigation of lysis centrifugation blood culture method for fungemia}; Yasuoka A et al.; We compared the lysis centrifugation blood culture system using Isolator (Du Pont) to the conventional blood culture bottle using BCB system (Roch), and the following results were obtained . In the investigation of experimental fungemia made by human blood and Candida albicans, the organism was detected 9/12 after 1 day and 11/12 after 3 days by Isolator, although the organism was detected 3/12 after 3 days and 11/12 after 6 days by the culture bottle system . In the experimental fungemia of Candida tropicalis, the organism was detected 12/12 after 2 days by Isolator, although it was detected 1/12 after 6 days and 4/12 after 10 days with the culture bottle system . Isolator showed faster and a more sensitive detection rate for fungemia . Clinical investigation of 94 cases (180 cultures) showed a 11.1% isolation rate with Isolator system and a 7.1% isolation rate with the culture bottle system . In 43 cases which were examined by the both systems, 7 cases were positive with Isolator, 3 positive with culture bottle . The culture bottle system isolated only Candida albicans and Candida parapsilosis, where as the Isolator isolated Candida albicans, Candida parapsilosis, Candida tropicalis, Candida krusei, and Candida spp . This investigation suggested that the Isolator system was an excellent method for the detection of fungemia.

Yeast, 1991 Jul, 7(5), 503 - 11
Assignment of most genes encoding major peroxisomal polypeptides to chromosomal band V of the asporogenic yeast Candida tropicalis; Kamiryo T et al.; The peroxisomes of the asporogenic yeast Candida tropicalis contain about 20 major polypeptides (PXPs) . We have isolated a number of genes encoding them; 11 POX genes encoded independent PXPs and three POY genes were likely to encode three other PXPs . To locate these genes on the chromosomes, chromosomes of C . tropicalis were separated by pulsed-field gel electrophoresis . Eight chromosomal bands were observed over the range of 1.0 Mbp (band 1) to 2.8 Mbp (band VIII); the genome size was estimated to be about 20 Mbp . Southern blot analysis showed that ten genes were on band V, three genes were on band IV, and the other gene was on band VI . Three genes gave hybridization signals of nearly equal intensity on two different chromosomal bands: POX6A and POX8B, on bands V and VII; and POX8A, on bands IV and VI . Ribosomal RNA genes also hybridized to two bands, VI and VII . Most genes assigned to only one band hybridized to two restriction fragments produced by either NotI or SfiI endonuclease . The results suggested that C . tropicalis was diploid and that restriction sites were conserved little between homologues . The three POX genes that were found on two chromosomal bands hybridized to not more than two restriction fragments, implying that the allelic genes were present on different chromosomal bands.

J Infect, 1991 Jul, 23(1), 77 - 80
Disseminated Candida tropicalis infection following endoscopic retrograde cholangiopancreatography; Ito M et al.; A necropsy case of a 59-year-old man who developed disseminated Candida tropicalis infection following endoscopic retrograde cholangiopancreatography (ERCP) is reported . Septic fungal infection complicating ERCP has not been described previously.

Mikrobiologiia, 1991 Jul-Aug, 60(4), 661 - 6
{Approximation of time-profiles of pH alteration by cells of Candida tropicalis by means of reactions of hypothetical linear object with negative feed-back}; Arzamastsev AA; The autostabilization of the environment pH value in batch cultures of yeast Candida tropicalis was experimentally investigated . The pH alternation had unidirectional compensatory nature, which allowed to represent the reaction of the system as the reaction of hypothetical linear object with negative feed-back . The transmission functions of structural elements of this object were determined . It was shown that such a model describes experimental data quite well in a wide range of initial environment pH values.

Eur J Clin Microbiol Infect Dis, 1991 Jul, 10(7), 588 - 92
In vitro comparison of cilofungin alone and in combination with other antifungal agents against clinical isolates of Candida species; Smith KR et al.; Cilofungin, a lipopeptide antifungal agent, was tested for in vitro activity alone and in combination with ketoconazole, itraconazole, flucytosine and amphotericin B against 102 clinical isolates of Candida species . At 48 hours all isolates of Candida albicans, Candida tropicalis, Candida paratropicalis and Candida glabrata were inhibited by less than or equal to 5 meg/ml of cilofungin . In contrast, the MIC90 for Candida krusei was 10 mcg/ml and for Candida parapsilosis greater than 40 mcg/ml . The interaction of combinations of cilofungin with amphotericin B, itraconazole, ketoconazole and flucytosine was additive or indifferent at 48 hours for 100%, 88%, 78% and 70% of all Candida species isolates, respectively . Overall, cilofungin demonstrated good activity in vitro against most Candida species isolates.

Oral Microbiol Immunol, 1991 Jun, 6(3), 187 - 90
Identification of oral yeasts by polyacrylamide gel electrophoresis; Maiden MF et al.; Protein profiles of sonicated cells for 9 species of yeast isolated from oral samples were obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and visualized with a silver stain . The profiles obtained on 12% acrylamide gels were distinct, and characteristic for 6 species of Candida, and Torulopsis glabrata, Yarrowia lipolytica, and Saccharomyces cerevisiae . Using this method, 79 fresh isolates of yeasts from saliva samples were identified; 58 as Candida albicans, 9 as Candida parapsilosis, 1 as Candida tropicalis, and 11 as T . glabrata . SDS-PAGE offers a rapid, convenient alternative or adjunct to yeast identification systems based on carbohydrate assimilation tests.

J Clin Microbiol, 1991 Jun, 29(6), 1089 - 94
Comparison study of broth macrodilution and microdilution antifungal susceptibility tests; Espinel-Ingroff A et al.; An evaluation of broth dilution antifungal susceptibility tests was performed by determining both the micro- and macrodilution MICs of amphotericin B, flucytosine, fluconazole, ketoconazole, and cilofungin against 38 isolates of Candida albicans, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, Cryptococcus neoformans, and Torulopsis glabrata . The following preliminary antifungal working group recommendations of the National Committee for Clinical Laboratory Standards for broth macrodilution tests with antifungal agents were used: inocula standardized to 1 x 10(4) to 5 x 10(4) CFU/ml with a spectrophotometer, RPMI 1640 medium buffered with morpholinopropanesulfonic acid (pH 7.0), incubation at 35 degrees C for 24 to 48 h, and an additive drug dilution procedure . Broth microdilution MICs were higher (two or more dilutions) than broth macrodilution MICs for all isolates tested with amphotericin B and for most isolates tested with ketoconazole, fluconazole, and cilofungin . MICs of flucytosine were the same by both techniques or lower by the broth microdilution test except in tests with C . neoformans . However, the only statistically significant differences between the two tests were observed with amphotericin B against all isolates (P = 0.01 to 0.07), ketoconazole against C . neoformans (P = 0.01 to 0.02), and cilofungin against C . albicans (P = 0.05 to 0.14) . Tests performed with less dense inocula (1 x 10(3) to 5 x 10(3} produced similar results.

J Clin Microbiol, 1991 Jun, 29(6), 1268 - 70
Restriction fragment analysis of a Candida tropicalis outbreak of sternal wound infections; Doebbeling BN et al.; An apparent single-source outbreak of Candida tropicalis sternal wound infections in eight patients was investigated by utilizing DNA restriction fragment analysis (RFA) with HindIII and BstNI . All eight outbreak isolates appeared to be identical and were easily differentiated from control isolates by DNA RFA . Compared with an arbitrarily selected reference outbreak isolate, greater than or equal to 95% of the bands in the restriction digests identified by a computerized image analysis system from each of the outbreak isolates were identical versus 13 to 53% of the bands in any of the nine control isolates . Outbreak strains were significantly more likely to match the reference outbreak isolate than were controls (P less than 0.0001) . The RFA was greatly facilitated by the use of computerized image analysis and confirmed the epidemiologic link between a scrub nurse and the infected patients.

Arch Biochem Biophys, 1991 May 1, 286(2), 504 - 10
The reductive half-reaction in acyl-CoA oxidase from Candida tropicalis: interaction with acyl-CoA analogues and an unusual thioesterase activity; Wang R et al.; A series of acyl-CoA analogues has been used to probe the substrate binding site and reductive half-reaction of acyl-CoA oxidase from the alkane utilizing yeast Candida tropicalis . Alkyl-SCoA thioethers, from octyl- to hexadecyl-SCoA, bind to the oxidase with progressively larger spectral perturbation of the flavin chromophore and with an incremental binding energy of about 260 cal/methylene group . The hydrocarbon binding subsite for acyl-CoA oxidase appears extensive and only weakly hydrophobic . CoA binding per se appears to contribute about 2.8 kcal to the observed binding energy . A number of acyl-CoA analogues such as 3-thia-acyl-, 3-oxa-acyl-, trans-3-enoyl-, and 3-keto-acyl-CoA derivatives form charge transfer complexes with the oxidase, but these long wavelength bands are both less pronounced and much less stable than those encountered with the acyl-CoA dehydrogenases . This instability reflects an intrinsic thioesterase activity of the oxidase which is observed with those ligands forming enolate to oxidized flavin charge-transfer complexes, but not with normal substrates such as palmitoyl-CoA . Chemical precedent suggests that these enzyme-bound enolates eliminate CoA via a ketene intermediate . The differences in behavior between acyl-CoA oxidase and dehydrogenase toward the ligands used in this work are discussed in terms of the need to exclude oxygen from productive encounters with substrate-reduced dehydrogenase.

Jpn J Antibiot, 1991 May, 44(5), 580 - 7
{In vitro antifungal activity of itraconazole: a comparative study with ketoconazole}; Hiratani T et al.; In vitro antifungal activities of itraconazole (ITZ), a new oral triazole antifungal agent, were studied against a wide range of medically important fungi including 16 genera, 37 species and 51 strains stocked in this center . The test was carried out using the agar dilution method on Sabouraud dextrose agar . ITZ showed equal or superior antifungal activities to ketoconazole against most strains of pathogenic yeasts, dimorphic fungi, non-pigmented hypomycetes, dermatophytes and dematiacious fungi . Although some strains of Candida albicans and Candida tropicalis were not completely inhibited by ITZ at concentrations up to 80 micrograms/ml, partial growth inhibitions were observed even at drug concentrations as low as 0.04 microgram/ml . The antifungal activity of ITZ against C . albicans was markedly influenced by medium composition, medium pH, inoculum size and incubation time.

Jpn J Antibiot, 1991 Mar, 44(3), 357 - 64
{Susceptibility to miconazole (base) of isolates from the oral cavity and esophagus of patients with mycosis}; Uchida K et al.; An antifungal agent of the imidazole class, miconazole (MCZ) (base), was investigated for its in vitro antifungal activities against isolates from the oral cavity and esophagus of patients with mycosis, and the following findings were obtained: 1 . Those yeast-like fungi which were presumed to be the causative agents were isolated from the oral cavity and esophagus of patients with mycosis, and identified . Candida albicans was the most frequently occurring species, accounting for 71.4% of all the isolates . Less frequently, several other Candida species, such as Candida krusei, Candida tropicalis, Candida lusitaniae and Candida lipolytica were also isolated mostly from patients with polymicrobial infections . 2 . The MICs of MCZ against the isolates of Candida species which were obtained from materials from patients with oral and esophageal candidiasis as the presumable causative agent distributed through such a wide range as less than or equal to 0.04 to 20 micrograms/ml, and susceptibilities of these isolates to MCZ proved to be no less than those to amphotericin B, which was used as the control drug . The results suggest that an appropriate preparation of MCZ (base) will be effective in treating mycosis of the oral cavity and esophagus.

Vopr Pitan, 1991 Mar-Apr, (2), 60 - 3
{Effect of a new exopolysaccharide on animal body}; Smoliar VI et al.; The influence of new exopolysaccharide, synthesized by Candida tropicalis and Pseudomonas sp . consortium in ethanol, on the animal body was studied in chronic experiments conducted in 200 non-inbred white rats . The animals were sacrificed after 3, 10 and 18 months of follow-up . The results of the investigations permit a conclusion that exopolysaccharide given to the animals during a long period does not produce a significant effect on the animal's growth and development, on the antitoxic function of the liver, on the parameters of protein, lipid and carbohydrate metabolism, on oxidizing phosphorylation in the mitochondria of the liver, and is low-hazard with respect to allergenic properties.

Can J Microbiol, 1991 Mar, 37(3), 226 - 32
Ammonium assimilation by Candida albicans and other yeasts: a 13N isotope study; Holmes AR et al.; Nuclear magnetic resonance spectra of cultures of Candida albicans incubated in the presence of 15N-labelled ammonium demonstrated that glutamine and glutamate were the only initial products of ammonium assimilation . The nature of the route of assimilation in the yeasts Candida albicans, Saccharomyces cerevisiae, and Candida tropicalis was further examined by the use of the short-lived isotope 13N . {13N}ammonium was generated in the reaction 16O(p,alpha)13N, induced by proton bombardment of water in tandem accelerator . High-pressure liquid chromatography was used to separate and identify the products of assimilation, and radioactivity was detected and corrected for decay, using a computer-linked NaI scintillation detector . In the three yeasts studied, the labelled ammonium was assimilated into the acid-extractable fraction of cell suspensions within 1 min, and over 75% was converted to glutamine and glutamate . Subsequent to exhaustion of the labelled ammonium, the stoichiometry of the distribution of radiolabel was consistent with a net transfer of radiolabel from glutamine to glutamate, confirming the operation of glutamate synthase (EC 1.4.1.14) in these yeasts . Initial assimilation of label was mostly into glutamine (at a maximal rate within 10 s in C . albicans), whereas accumulation in glutamate did not occur at maximal rate until more than 70% of the labelled ammonium had been assimilated (between 30 and 60 s in C . albicans) . We conclude that the glutamine synthetase-glutamate synthase pathway is the major route of ammonium assimilation in C . albicans and also in nitrogen-starved cultures of S . cerevisiae and C . tropicalis.

Chem Pharm Bull (Tokyo), 1991 Feb, 39(2), 493 - 5
Synthesis of Mannich bases of 5-hydroxynapthalene-1,8-carbolactone as potential antifungal or antitumor agents; Fillion H et al.; Mannich bases of 5-hydroxynaphthalene-1,8-carbolactone 1 were prepared from various secondary amines or bulky primary amines and formaldehyde . They were isolated in almost all cases as hydrochlorides . These derivatives were submitted to in vitro antifungal and cytotoxic assays . The antifungal assays were performed against three strains of yeasts and five strains of human pathogenic fungi . Two of the tested compounds, 2i and 2j, exhibited interesting antifungal activities against Candida albicans and Candida tropicalis . The cytotoxic activity was evaluated towards L 1210 leukemia cells . Almost all of the Mannich bases had shown significant activity against this tumor cell line as values of IC50 less than or equal to 4 micrograms/ml are considered interesting . Only one derivative 2 developed better cytotoxicity than the parent compound 1.

Mycopathologia, 1991 Feb, 113(2), 81 - 7
Phagocyte-mediated killing of Candida tropicalis; Lindemann RA et al.; Human peripheral monocytes (MO), neutrophils (PMN), and lymphocytes (PBL) were tested for their ability to kill Candida tropicalis . With incubation times between 30 min and 2 h, unstimulated MO and PMN, but not PBL, were efficient killers of C . tropicalis . Both leukocyte subsets were able to kill at minimum 2.5:1 effector to target ratios . Pre-incubation of MO for 24 h with interferon-gamma or tumor necrosis factor (TNF) increased their ability to kill yeast targets . TNF alone had no effect on C . tropicalis targets at concentrations up to 1000 U/ml . PBL activated for 4 d with interleukin-2 did not kill yeast targets . PMN exhibited more cytocidal efficiency per cell than MO in these assays . Direct contact of effectors and targets was required; no significant killing by PMN or MO supernatants was measured . PMN-mediated killing, but not MO killing, was inhibited by a mixture of catalase and superoxide dismutase suggesting that oxygen-dependent killing mechanisms were partially responsible for candidacidal activity.

J Gen Microbiol, 1991 Jan, 137 ( Pt 1), 161 - 7
Variation of colony morphology and chromosomal rearrangement in Candida tropicalis pK233; Suzuki T et al.; UV irradiation treatment of the asexual yeast Candida tropicalis gave rise to morphological mutants exhibiting at least four different types of abnormal colonies on glucose-containing solid medium . These mutants were named according to their colony morphologies: 'doughnut', 'frilly', 'echinoid' and 'walnut' mutants . The doughnut mutant produced a wrinkled colony with a hollow in its central region that was rich in filamentous pseudohyphal cells . With increased incubation time, the colony gradually changed to a reticulate shape . The parent strain, which normally produced smooth colonies, gave similar colonies to those of the doughnut mutant when grown in medium containing oleic acid as carbon source . Both the frilly and the walnut mutants produced pseudohyphal cells in a similar fashion to the doughnut mutant . The echinoid mutant produced an echinulate colony morphology with aerial hyphae and contained true hyphal cells as well as pseudohyphal ones . Pulsed-field gel electrophoresis showed that the echinoid and frilly mutants had different karyotypes from that of their parent strain, suggesting the occurrence of chromosomal rearrangements associated with these morphological mutations.

Chemotherapy, 1991, 37(1), 23 - 31
In vitro susceptibility of 119 yeast isolates to fluconazole, 5-fluorocytosine, amphotericin B and ketoconazole; Morace G et al.; The in vitro activity of fluconazole was tested against 13 yeast species (119 strains) isolated from clinical specimens during a 3-month period . For comparative purposes, three other antifungal compounds (5-fluorocytosine, amphotericin B and ketoconazole) were also tested . The tests were carried out using a microautomated method previously developed in our laboratory . The method allowed us to determine the minimum inhibitory concentration (MIC) of the four antifungal drugs used . For each of the drugs we utilized different media . The MIC ranges (mg/l) of fluconazole were: 0.04-12.5 for Candida albicans, 0.19-6.25 for Candida parapsilosis, 12.5-50 for Candida krusei, 0.04-100 for Candida tropicalis, 0.04- greater than 100 for Candida glabrata, 0.09- greater than 25 for Cryptococcus neoformans, 0.09-0.78 for Saccharomyces cerevisiae, 6.25- greater than 100 for Trichosporon beigelii and 0.09-0.19 for Blastoschizomyces capitatus (Trichosporon capitatum) . The MIC value (mg/l) was 0.39 for Candida guilliermondii and Candida lusitaniae, greater than 100 for Cryptococcus laurentii and 0.09 for the 3 isolates of Torulopsis candida . These results were obtained using the medium recommended for in vitro testing of fluconazole (high-resolution medium) by Pfizer UK.

J Med Vet Mycol, 1991, 29(2), 93 - 8
Agar disk diffusion susceptibility tests with cilofungin (LY 121019); Espinel-Ingroff A et al.; Cilofungin (LY 121019) is a semi-biosynthetic lipopeptide agent which is active both in vitro and in vivo against isolates of Candida species . The purpose of this study was to demonstrate the usefulness of experimental 10- and 20-micrograms cilofungin disks in predicting probable in vitro susceptibility to this drug . Fifty-two isolates of pathogenic yeasts, which included 34 isolates of Candida albicans, 10 isolates of Candida tropicalis and eight isolates of Candida glabrata, were tested . Both agar dilution minimal inhibitory concentration (MIC) tests and matched agar disk diffusion tests were performed using 14-h-old broth cultures grown in single strength yeast nitrogen base (YNB) supplemented with asparagine and dextrose . Both tests were incubated at 35 degrees C and the results were read after 24 h . Regression analysis was used to measure the degree of correlation between MIC values and matched averaged zones of inhibition, and demonstrated that both the 10-micrograms (r = 0.9278) and 20-micrograms (r = 0.9082) disks can predict probable MIC values.

Arch Microbiol, 1991, 156(6), 439 - 43
High level expression of isocitrate lyase gene of n-alkane-utilizing yeast Candida tropicalis in Saccharomyces cerevisiae; Oda K et al.; The genomic DNA of peroxisomal isocitrate lyase (ICL) isolated from an n-alkane-assimilating yeast, Candida tropicalis, was truncated to utilize the original open reading frame under the control of the GAL7 promoter and was expressed in Saccharomyces cerevisiae . The recombinant ICL was synthesized as a functionally active enzyme with a specific activity similar to the enzyme purified from C . tropicalis, and was accounted for approximately 30% of the total extractable proteins in the yeast cells . This recombinant enzyme was easily purified to homogeneity . N-Terminal amino acid sequence, molecular masses of native form and subunit, amino acid composition, peptide maps, and kinetic parameters of the recombinant ICL were essentially the same as those of ICL purified from C . tropicalis . From these facts, S . cerevisiae was suggested to be an excellent micro-organism to highly express the genes encoding peroxisomal proteins of C . tropicalis.

Biol Met, 1991, 4(4), 233 - 7
Biosorption of copper by yeasts; Junghans K et al.; The ability to accumulate copper from aqueous solutions was determined with different yeast species . Yeast cells did not show any significant differences in process kinetics . The uptake was very fast and was influenced by environmental factors . The metal-accumulating capacity differed among the tested strains . The yeast Candida tropicalis and Pichia guilliermondii were chosen for extensive research . Cells of the stationary growth phase were able to adsorb a high amount of copper . The uptake capacity decreased with increasing biomass concentration . Copper adsorption obeyed the Freundlich isotherm . Optimal pH range was between 5 and 7 . The biomass could be used repeatedly for biosorption after desorption by mineral acids.

Int Arch Allergy Appl Immunol, 1991, 96(2), 142 - 8
Characterization of a monoclonal antibody (RJ5) against the immunodominant 41-kD antigen of Candida albicans; Shen HD et al.; A 41-kD component of Candida albicans was identified to be the major antigen radioimmunoprecipitated by antibodies with increased titers in the sera of patients with invasive candidiasis . A mouse monoclonal antibody (RJ5) was generated which, by immunoblotting, showed positive reactivity to the immunoprecipitated 41-kD component . By two-dimensional gel electrophoresis and immunoblotting, MoAb RJ5 was shown to react with different isoforms of the 41-kD component with pI values from 6.1 to 6.9 . Furthermore, MoAb RJ5 showed positive reactivity to cytoplasmic antigens of C . albicans by frozen section and immunoperoxidase staining . By SDS-polyacrylamide gel electrophoresis and immunoblotting, MoAb RJ5 showed no cross-reactivity to antigens of Candida tropicalis and Candida parapsilosis . The epitope of the 41-kD molecule recognized by MoAb RJ5 was susceptible to treatment of proteinase K at concentrations of greater than or equal to 5 micrograms/ml, and was relatively resistant to periodate oxidation with concentration of NaIO4 up to 20 mM . This MoAb may be useful in the purification and characterization of the immunodominant 41-kD antigen of C . albicans, and as a probe in the detection of Candida antigens in the sera of patients with invasive candidiasis.

Chem Pharm Bull (Tokyo), 1990 Dec, 38(12), 3449 - 51
Reduction of nitroolefin using microorganisms; Mori A et al.; Microbial reduction of 1-phenyl-2-nitro-1-propene (3) was carried out using 57 strains of yeast, 40 strains of aerobic and facultatively anaerobic bacteria and 40 strains of strictly anaerobic bacteria . Nine strains of yeast (Candida tropicalis, etc.,) had the ability to reduce (3) to 1-phenyl-2-nitropropane (1) (94.1%-60.3% yield) . The ability of the aerobic and anaerobic bacteria was weaker than yeast (35.6%-14.0% and less than 5%, respectively) . When 11 strains of strictly anaerobic bacteria (Clostridium innocuum etc.,) were used, a final reduced product like amphetamine (2) was detected, although the efficiency of reduction was very poor.

Appl Environ Microbiol, 1990 Dec, 56(12), 3785 - 92
Oxygen requirements of yeasts; Visser W et al.; Type species of 75 yeast genera were examined for their ability to grow anaerobically in complex and mineral media . To define anaerobic conditions, we added a redox indicator, resazurin, to the media to determine low redox potentials . All strains tested were capable of fermenting glucose to ethanol in oxygen-limited shake-flask cultures, even those of species generally regarded as nonfermentative . However, only 23% of the yeast species tested grew under anaerobic conditions . A comparative study with a number of selected strains revealed that Saccharomyces cerevisiae stands out as a yeast capable of rapid growth at low redox potentials . Other yeasts, such as Torulaspora delbrueckii and Candida tropicalis, grew poorly mu max, 0.03 and 0.05 h-1, respectively) under anaerobic conditions in mineral medium supplemented with Tween 80 and ergosterol . The latter organisms grew rapidly under oxygen limitation and then displayed a high rate of alcoholic fermentation . It can be concluded that these yeasts have hitherto-unidentified oxygen requirements for growth.

Eur J Clin Microbiol Infect Dis, 1990 Oct, 9(10), 738 - 44
Activity of amphotericin B and intraconazole against intraphagocytic Candida albicans; Ponce E et al.; The activity of amphotericin B and intraconazole against intracellular Candida albicans was determined in vitro using murine resident peritoneal macrophages . Amphotericin B at concentrations of 0.5 and 2 micrograms/ml produced significantly less rapid killing of intracellular than of extracellular Candida albicans as measured in macrophage-free medium . Amphotericin B at concentrations of 0.1 micrograms/ml or itraconazole concentrations of up to 3 micrograms/ml produced only fungistatic or limited fungicidal activity against both intracellular and extracellular organisms . Against intracellular Candida albicans amphotericin B acted by direct antifungal action rather than through stimulation of macrophage function, as demonstrated by the fact that (i) activity persisted when macrophages were successively exposed to amphotericin B, washed and disrupted by sonication, and (ii) no activity was seen when amphotericin B was tested against intracellular amphotericin B-resistant Candida tropicalis or Salmonella typhimurium . Pre-exposure of macrophages to itraconazole (0.4 micrograms/ml) inhibited subsequent killing activity of amphotericin B (2 micrograms/ml) against intracellular susceptible Candida albicans . These experiments validate the conventional methods of susceptibility testing for determining the fungistatic activity of antifungal agents.

J Biol Chem, 1990 Sep 25, 265(27), 16428 - 36
Isolation and characterization of the alkane-inducible NADPH-cytochrome P-450 oxidoreductase gene from Candida tropicalis . Identification of invariant residues within similar amino acid sequences of divergent flavoproteins; Sutter TR et al.; The gene coding for the Candida tropicalis NADPH-cytochrome P-450 oxidoreductase (CPR, NADPH: ferricytochrome oxidoreductase, EC 1.6.2.4) was isolated by immunoscreening of a C . tropicalis lambda gt11 expression library and colony hybridization of a C . tropicalis genomic library . The C . tropicalis CPR gene produces a 2.35-kilobase mRNA transcript, levels of which were shown to be increased 16-fold in cells grown on tetradecane relative to cells grown on glucose as the sole carbon source . A 3-kilobase DNA fragment was sequenced, including 554 and 397 base pairs of 5'- and 3'-noncoding sequence, respectively . A single open reading frame of 2040 base pairs was identified and predicts a 76,683-Da polypeptide of 680 amino acid residues . The deduced C . tropicalis CPR amino acid sequence was compared with each of the CPR sequences reported from other organisms and invariant residues were identified . Multiple pairwise alignments of divergent members of protein families, previously recognized for their sequence similarities in their respective binding domains for FMN, FAD, and NADPH, have allowed identification of a subset of these invariant residues . From these analyses we infer the importance of 25 of the 680 amino acid residues.

FASEB J, 1990 Sep, 4(12), 3028 - 32
A common ancestor for Candida tropicalis and dehydrogenases that synthesize antibiotics and steroids; Baker ME; Candida tropicalis peroxisomes contain a 905-residue trifunctional enzyme with hydratase-dehydrogenase-epimerase activity that is important in fatty acid beta-oxidation . At its amino terminus are two tandem copies of an approximately 280 residue domain of unknown function . We provide evidence that this domain is homologous to oxidoreductases used for metabolizing sugars and synthesizing antibiotics and steroids such as estradiol, androstenedione, corticosterone, and hydrocortisone . The trifunctional enzyme shows no sequence similarity to the bifunctional hydratase-dehydrogenase found in animal peroxisomes and plant glyoxysomes, which are homologs of each other . We suggest that the C . tropicalis trifunctional enzyme and the animal and plant bifunctional enzymes have different ancestors.

Eur J Clin Microbiol Infect Dis, 1990 Sep, 9(9), 697 - 9
In vitro activity of the new semi-synthetic polypeptide cilofungin (LY121019) against Aspergillus and Candida species; Huang A et al.; The in vitro activity of cilofungin (LY121019), a new semi-synthetic antifungal agent was evaluated . Potent activity was seen against Candida albicans and Candida tropicalis, with almost identical MIC and MFC results, whereas no activity was seen against any isolates of Candida parapsilosis or three Aspergillus spp . However, MICs were dependent on medium and test conditions chosen . It is concluded that cilofungin has good activity against some medically important yeasts in vitro, and that its in vitro activity depends on the method used.

Haematologica, 1990 Sep-Oct, 75(5), 480 - 1
An unusual case of Candida tropicalis sepsis in a patient submitted to allogeneic bone marrow transplantation; Rosti G et al.; We describe an exceptional case of Candida tropicalis sepsis in a patient submitted to allogeneic BMT; the diagnosis was made on a peripheral blood smear, when the pt was neutropenic and only mildly febrile . The combination of GM-CSF to accelerate hematological recovery and the possibility of administering large doses of a liposomal form of Amphotericin B were the contributing factors to the resolution of the infection.

Rinsho Byori, 1990 Aug, 38(8), 931 - 6
{Identification and susceptibility of clinically isolated yeast-like fungi}; Fukuchi K et al.; We studied the identification and susceptibility of clinically isolated yeast-like fungi at Showa University Hospital from April 1988 to March 1989 . Clinically significant of yeast-like fungi were observed in 7.1% of specimens from outpatients, 13.0% of inpatients . In both outpatients and inpatients, yeast-like fungi were isolated mainly from sputum and urine . But, one third of them were considered as non-pathogenic and not identified . The species of isolates were, Candida albicans 57%, Candida tropicalis 14% and Candida glabrata 8% in both inpatients and outpatients, and these species shared most part . The isolation frequency of Candida parapsilosis was higher in blood and cerebrospinal fluid (CSF) specimen than the others . The susceptibility test by agar dilution method indicated most of the isolates were susceptible to Amphotericin B and Miconazole (MIC less than or equal to 25 micrograms/ml) . There was no difference in MIC between predominantly isolated fungi and commonly isolated fungi . Notably, isolates from blood and CSF showed a significant high tolerance against Amphotericin B and Miconazole than from the other specimens . The MICs of Fluconazole were shown to be very high (greater than 100 micrograms/ml) in normal Sabouraud agar, were decreased dose-dependently by human sera in the medium . These findings indicated the component(s) of sera enhanced the anti-fungal activity of Fluconazole.

Appl Environ Microbiol, 1990 Aug, 56(8), 2562 - 4
Isolation of Candida tropicalis auxotrophic mutants; Gleeson MA et al.; An enrichment scheme using nystatin was designed for the isolation of auxotrophic mutants from the diploid-alkane-utilizing yeast Candida tropicalis . A collection of 194 auxotrophs representing 7 phenotypes was isolated . One class of mutants was identified as having a defect in histidinol dehydrogenase activity and a second class of mutants was identified as having a defect in orotidine monophosphate decarboxylase activity . These strains are good candidates to be carrying mutations corresponding to the HIS4 and URA3 genes of Saccharomyces cerevisiae.

J Bacteriol, 1990 Aug, 172(8), 4571 - 7
Development of an integrative DNA transformation system for the yeast Candida tropicalis; Haas LO et al.; We developed the alkane and fatty-acid utilizing yeast Candida tropicalis as a host for DNA transformations . The system is based on an auxotrophic mutant host of C . tropicalis which is defective in orotidine monophosphate decarboxylase (ura3) . The ura3 host was isolated by mutagenesis and a double-selection procedure that combined nystatin enrichment selection and 5-fluoro-orotic acid resistance selection . As a selectable marker, we isolated and characterized the C . tropicalis URA3 gene . Plasmid vectors that contained the C . tropicalis URA3 gene transformed the C . tropicalis mutant host at a frequency of 10(3) to 10(4) transformants per micrograms of plasmid DNA . Vectors that contained the Saccharomyces cerevisiae URA3 gene could not transform C . tropicalis . DNA transfer was accomplished by modified versions of either spheroplast generation (CaCl2-polyethylene glycol)-fusion or cation (LiCl) procedures developed for S . cerevisiae . Plasmid vectors that had been cut within the C . tropicalis URA3 fragment integrated by homologous recombination at the URA3 locus.

Infect Immun, 1990 Aug, 58(8), 2613 - 20
Characterization of Candida albicans mannan-induced, mannan-specific delayed hypersensitivity suppressor cells; Garner RE et al.; We have shown previously that CBA/J mice immunized with Candida albicans developed delayed hypersensitivity (DH) demonstrable with mannan (MAN) extracted from the same organism and that the intravenous (i.v.) injection of MAN prior to or during the immunization phase resulted in the suppression of the MAN-specific DH response . In this study, we demonstrate that MAN-induced suppression of DH is a T-lymphocyte-mediated phenomenon . Suppressor cells induced in vivo by the i.v . injection of MAN into naive mice 1 to 7 days prior to harvest were passaged through nylon wool, treated with various surface-specific antibodies and complement, and then injected i.v . into immunized syngeneic recipients . Enrichment of splenic T cells by passage over nylon wool and transfer of the nylon-wool-nonadherent populations to immunized recipient mice suppressed DH in a dose-dependent manner . Depletion of Thy+ or Lyt-2+ cells from nylon-wool-nonadherent populations regularly ablated the ability of such suspensions to transfer suppression . Treatment of the same transfer suspensions with anti-Lyt-1 had variable effects, suggesting that the surface density of the Lyt-1 antigen was not as constant from population to population as was the Lyt-2 antigen . In addition, C . albicans MAN-induced suppressor cells were able to suppress DH demonstrable with Candida tropicalis MAN in animals immunized with C . tropicalis . Suppression of DH by MAN in this model, therefore, is mediated by Thy+ Lyt-2+ lymphocytes.

J Clin Microbiol, 1990 Jul, 28(7), 1509 - 13
Discriminant analysis of cellular fatty acids of Candida species, Torulopsis glabrata, and Cryptococcus neoformans determined by gas-liquid chromatography; Marumo K et al.; We used discriminant analysis of cellular fatty acid compositions determined by gas-liquid chromatography to differentiate yeastlike fungi (a total of 190 strains; including 37 Candida albicans strains, 21 Candida krusei strains, 13 Candida guilliermondii strains, 37 Candida tropicalis strains, 10 Candida pseudotropicalis strains, 24 Candida parapsilosis strains, 32 Torulopsis glabrata strains, and 16 Cryptococcus neoformans strains) . Previous results with a standard strain of C . albicans indicated that reproducible fatty acid chromatograms can be obtained with cells grown in a medium of 2% Sabouraud glucose agar at 35 degrees C for between 48 and 72 h . These conditions were also maintained in cultures of the other organisms that we studied . The cellular fatty acid compositions of the organisms were determined quantitatively by gas-liquid chromatography and analyzed by discriminant analysis . The total correct identification expressed as relative peak percent was 95.8% (89.2% for C . albicans to 100% for C . krusei, C . guilliermondii, C . pseudotropicalis, T . glabrata, and C . neoformans) . The total correct identification expressed as the common peak (palmitic acid) ratio was 94.7% (87.5% for C . parapsilosis to 100% for C . pseudotropicalis, T . glabrata, and C . neoformans) . Both results suggest that cellular fatty acid compositions can be differentiated by this method.

Infect Immun, 1990 Jul, 58(7), 2105 - 14
Production and characterization of monoclonal antibodies to cell wall antigens of Aspergillus fumigatus; Ste-Marie L et al.; Two murine monoclonal antibodies (MAbs) against Aspergillus fumigatus were produced and characterized . Splenocytes from cell wall-immunized BALB/c mice were fused with SP2/0 myeloma cells . The hybridomas were screened with a cold alkali (CA) extract of mycelium containing protein, mannose, and galactose, and two MAbs of the immunoglobulin M class were purified from ascites fluid . MAbs 1 and 40 were characterized by double immunodiffusion against CA antigen, indirect enzyme immunoassay with mannans of Candida albicans serotypes A or B or Candida tropicalis, indirect immunofluorescence with C . albicans- or A . fumigatus-infected tissues, indirect immunofluorescence with smears of other pathogenic fungi, Western blotting (immunoblotting) with the lectin concanavalin A or BS-1 from the seeds of Bandeirea simplicifolia, and immunoelectron microscopy . MAb 1 did not cross-react with Candida mannan and recognized a periodate-sensitive, pronase- and heat-resistant epitope in CA antigen and three mannose- and galactose-containing components (80, 62, and 49 kilodaltons) of a mycelial homogenate . Immunoelectron microscopy demonstrated binding of MAb 1 to the inner cell wall and intracellular membranes of hyphae and conidia of A . fumigatus . Circulating antigen was detected in experimental invasive aspergillosis by inhibition enzyme immunoassay with MAb 1 and CA antigen . MAb 40 was a nonprecipitating antibody cross-reactive with Candida species, and competition for an epitope located diffusely in the cell wall of A . fumigatus hyphae was demonstrated by incubating MAb 40 with mannan of C . albicans serotype A . These results suggest that MAb 1 recognizes immunodominant oligogalactoside side chains of A . fumigatus galactomannan, while MAb 40 binds to mannopyranosyl side chains common to A . fumigatus galactomannan and C . albicans mannan.

Curr Genet, 1990 Jun, 17(6), 481 - 6
In vivo import of Candida tropicalis hydratase-dehydrogenase-epimerase into peroxisomes of Candida albicans; Aitchison JD et al.; We present a system for studying peroxisomal protein targeting in Candida . We have expressed the Candida tropicalis gene encoding hydratase-dehydrogenase-epimerase (HDE) in Candida albicans . Immunoblot analyses of C . albicans transformants demonstrate the presence of oleic-acid inducible HDE (100 kDa) in peroxisomes of transformed cells, but not of control cells . Peroxisomes isolated from transformed cells show increased beta-hydroxyacyl-CoA dehydrogenase specific activity, indicating that HDE is imported into peroxisomes of C . albicans where it is enzymatically active . C . albicans provides a useful model for the study of protein targeting to peroxisomes in vivo.

Oral Surg Oral Med Oral Pathol, 1990 Jun, 69(6), 683 - 7
Characterization of the mycoflora from oral mucosal surfaces of some HIV-infected patients; Franker CK et al.; Oral mucosal surfaces from 54 patients seropositive for human immunodeficiency virus (HIV) were assayed for the presence of cultivable yeasts . Oral colonization with Candida albicans, represented by 6 biotypes, was evident in 35 persons . The closely related variant, Candida stellatoidea, was found in 3 patients . Single isolates of Candida parapsilosis, Candida glabrata, Candida tropicalis, and Candida paratropicalis were also identified . One patient harbored a population of Saccharomyces cerevisiae . The susceptibilities of these 43 isolates to clotrimazole and nystatin were compared by the disk diffusion technique.

Pathol Biol (Paris), 1990 Jun, 38(5 ( Pt 2)), 575 - 8
{Comparative sensitivity of yeasts to ketoconazole, itraconazole and fluconazole using a liquid medium standardized micromethod}; Guinet R et al.; The minimal inhibitory concentrations of 200 yeast strains (48 reference strains and 152 recently isolated from pathological products) were evaluated with a new standardized micromethod using a liquid medium comparatively for ketoconazole, itraconazole and fluconazole . The ready-to-use microtitration plates contained the antifungal agents in concentrations ranging from 100 to 0.10 mg/l . The cumulative MIC curves clearly showed the superiority of ketoconazole, fluconazole being the less active product . These results were confirmed after categorization in sensitive (S: MIC less than 0.78 mg/l), intermediate (I: 0.78 less than MIC less than 6.25) or resistant (R: MIC greater than 6.25) . These results are discussed particularly for fluconazole showing the largest half-life and for which higher plasmatic levels could be achieved . Important variations were observed depending on the species and as example for Candida tropicalis the sensitivities were ketoconazole 100%, itraconazole 55% and fluconazole 0% . The less sensitive species to the 3 azoles were Saccharomyces cerevisiae and Torulopsis glabrata . This new micromethod being very easy to use and allowing the determination of fungicidal activities should be introduced in routine.

Infect Immun, 1990 Jun, 58(6), 2005 - 7
The spermicidal compound nonoxynol-9 increases adhesion of Candida species to human epithelial cells in vitro; McGroarty JA et al.; All 25 strains of Candida spp . tested were able to grow in medium supplemented with 25% nonoxynol-9 in vitro . Adhesion of Candida spp . to HeLa cells was found to increase in the presence of 5% nonoxynol-9 (2.2- to 6.6-fold; P less than 0.001) and, to a lesser extent, in 12.5% nonoxynol-9 . Adhesion of Candida strains cultured in medium supplemented with nonoxynol-9 varied, with five of six strains of Candida albicans and the single strain of Candida tropicalis demonstrating increases of 1.4 to 4.0 times control levels (P less than 0.05) . The increased adhesion did not correlate with altered yeast cell surface hydrophobicity or germ tube formation.

Eur J Biochem, 1990 May 31, 190(1), 107 - 12
A novel peroxisomal nonspecific lipid-transfer protein from Candida tropicalis . Gene structure, purification and possible role in beta-oxidation; Tan H et al.; We have sequenced the nucleotides of the gene POX18 that encodes PXP-18, a major peroxisomal polypeptide inducible by oleic acid in the yeast Candida tropicalis . POX18 had a single open reading frame of 127 amino acids . Some 33% of the amino acid sequence of the predicted basic polypeptide (13,805 Da), was identical to that of the nonspecific lipid-transfer protein (sterol carrier protein 2) from rat liver . PXP-18, purified to near homogeneity from isolated peroxisomes, had an amino-terminal sequence identical to that of the predicted polypeptide except for the initiator methionine, and had nonspecific lipid-transfer activity comparable to that of its mammalian equivalents . Unexpectedly, PXP-18 lacked the cysteine residue thought to be essential for the activity of this protein in mammals . RNA blot analysis showed that the POX18 gene was expressed exclusively in cells grown on oleic acid, suggesting that PXP-18 has a role in the beta-oxidation of long-chain fatty acids . PXP-18 modulated acyl-coenzyme A oxidase activity at low pH.

Kansenshogaku Zasshi, 1990 May, 64(5), 636 - 41
{A successfully treated case of infective endocarditis due to Candida tropicalis}; Kasugai H et al.; A 34-year-old man, a heavy drinker, was admitted with a high fever and hematuria two months previously . Surgery was performed for acute sever pancreatitis and postoperatively antibiotics were administered with intravenous hyperalimentation . After discharge he was readmitted and infective endocarditis was strongly suspected because of high fever, hematuria, Osler's nodes, Janeway's lesions, splinter hemorrhages and mitral regurgitation . Penicillin G in combination with Gentamycine therapy was started on the first hospital day . On the second hospital day, blood culture revealed Candida tropicalis so Miconazole therapy was commenced . On the forth hospital day, he underwent surgery for replacement of a mitral prosthesis with a prosthetic valve because he had embolus in the radial artery . Despite intensive antifungal therapy, he showed no improvement in clinical symptoms . Then we changed the antifungal drug from Miconazole to Amphotericin B and 5-fluorocytosine . On the 109th hospital day, his clinical symptoms improved . Antifungal therapy was halted and at present 10 months later, he is healthy.

Antimicrob Agents Chemother, 1990 May, 34(5), 875 - 80
Enzyme immunoconjugates utilizing glucose oxidase and myeloperoxidase are cytotoxic to Candida tropicalis; Casentini-Borocz D et al.; A dual-enzyme immunoconjugate system was evaluated for its cytotoxic effect on Candida tropicalis . Glucose oxidase, which generates hydrogen peroxide in the presence of glucose and oxygen and myeloperoxidase, which catalyzes the oxidation of halides in the presence of hydrogen peroxide, were each conjugated to a C . tropicalis-specific monoclonal antibody . Neither the glucose oxidase nor the myeloperoxidase conjugates exhibited any significant cytotoxic effect by themselves . A combination of glucose oxidase conjugate (3.2 ng/ml) and myeloperoxidase conjugate (12.8 ng/ml) in the presence of 5 mg of glucose per ml, 150 mM chloride, and 50 microM iodide was cytotoxic to C . tropicalis, killing 99.9% of the treated sample . Flow cytometry was used to characterize the binding of the conjugates to yeast cells and demonstrated that the binding of both conjugates to the yeast cell surface is required for cytotoxicity . In addition, the concentrations of conjugates required for a cytotoxic effect were below the concentrations required to saturate all of the yeast cell surface antibody-binding sites.

Gene, 1990 Apr 16, 88(2), 247 - 52
Structure and transcriptional control of the Saccharomyces cerevisiae POX1 gene encoding acyl-coenzyme A oxidase; Dmochowska A et al.; We have cloned the Saccharomyces cerevisiae gene coding for the peroxisomal enzyme: fatty acyl-CoA oxidase (POX) . The gene (named POX1) is unique in S . cerevisiae and has been identified through homology with the POX4 and POX5 genes of Candida tropicalis . The POX1 gene encodes a 84-kDa POX protein composed of 748 amino acids . The identity between the S . cerevisiae and C . tropicalis enzymes is about 40%, and there is a greater degree of similarity between the N termini than the C termini . A disruption of the POX1 coding sequence diminishes the ability of yeast cells to grow on oleic acid as a sole carbon source . The expression of the POX1 gene is regulated at the level of transcription, and is induced more than 25-fold by the addition of oleic acid to the medium.

Biochem Biophys Res Commun, 1990 Mar 30, 167(3), 1050 - 6
Specific, high-affinity binding sites for human luteinizing hormone (hLH) and human chorionic gonadotrophin (hCG) in Candida species; Bramley TA et al.; The presence of specific binding sites for {125I}-labelled hLH and hCG is described in Candida species . Binding was present in three strains of Candida albicans, and in Candida tropicalis, and was greatest in microsomes, though binding was also present in cytosol fractions . hLH and hCG mutually competed for these binding sites . Other hormones did not bind and did not compete for hLH binding sites . Scatchard plots showed two classes of binding sites, one with high affinity, low capacity and the other with lower affinity, high capacity binding in both microsomes and cytosol . This is the first report of specific binding sites for mammalian peptide hormones in a yeast.

J Cell Sci, 1990 Mar, 95 ( Pt 3), 463 - 70
Isolation and characterization of membranes from oleic acid-induced peroxisomes of Candida tropicalis; Nuttley WM et al.; We report a methodology for the isolation of peroxisome membranes from the yeast Candida tropicalis pK233 grown on oleic acid, and the characterization of the polypeptide and lipid compositions of these membranes . Peroxisomes purified in either sucrose or Nycodenz gradients are treated with Tris-HCl (pH 8.5) and then with sodium carbonate (pH 11.5) to yield a final peroxisome membrane preparation (hereafter called 'peroxisome membranes') . Electron microscopy revealed peroxisome membranes that are approximately 8.1 nm thick, have a typical trilaminar appearance, and form either flattened sheets or whorled structures . Peroxisome membranes contain 3.1% and 2.2% of the total protein of sucrose- and Nycodenz-gradient-purified peroxisomes, respectively . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed three predominant polypeptide bands of 34 (PMP 34), 29 (PMP 29), and 24 (PMP 24) x 10(3) Mr in peroxisome membranes . Immunoblotting with an antiserum to PMP 24 showed that PMP 24 segregates with the peroxisome membrane fractions and is induced by growth of Candida tropicalis on oleic acid . Peroxisome membranes contain neutral lipids and phospholipids . The principal phospholipids are phosphatidyl choline and phosphatidyl ethanolamine . The phospholipid/protein ratio of peroxisome membranes is approximately 430 nmol mg-1.

Genetika, 1990 Mar, 26(3), 567 - 9
{The role of lipids during exposure of mutant cells of Candida tropicalis D-2 to heterologous DNA after treatment with calcium cations}; Danilenko II et al.; It was shown that treatment of mutant cells of Candida tropicalis D-2 strain with deficiency for histidine and respiration Ca2+ ions leads to increase in the number of revertants at the action of heterologous DNA . The effect of preliminary treatment with Ca2+ ions depends on the time of their action, the number of intracellular lipids, the nature of DNA and temperature of treatment.

J Biochem (Tokyo), 1990 Feb, 107(2), 262 - 6
Peroxisomal isocitrate lyase of the n-alkane-assimilating yeast Candida tropicalis: gene analysis and characterization; Atomi H et al.; A genomic DNA clone encoding isocitrate lyase, a key enzyme of the glyoxylate cycle and a peroxisomal enzyme of the n-alkane-assimilating yeast Candida tropicalis has been isolated with a cDNA probe from the yeast lambda EMBL library . Nucleotide sequence analysis of the genomic DNA clone disclosed that the region coding isocitrate lyase had a length of 1,650 base pairs, corresponding to 550 amino acids (61,602 Da) . RNA blot analysis demonstrated that only one kind of mRNA (2 kb) supposed to be transcribed from this gene was present in the cells . A comparison of the amino acid sequences was made with the isocitrate lyase of castor bean, one of the glyoxysomal enzymes, and the enzyme of E . coli . The isocitrate lyases of C . tropicalis and castor bean had high homology, and the presence of some amino acid stretches conserved in all three enzymes suggests that these might be involved in the catalysis of the common reaction . There was an insertion common to the isocitrate lyases of C . tropicalis and castor bean, which is of interest concerning their evolution . In the C-terminal region, a characteristic sequence similar to that previously proposed as the import signal to peroxisomes was present.

Am J Vet Res, 1990 Jan, 51(1), 71 - 5
Disassociation of bactericidal and fungistatic activities from the oxidative burst of avian macrophages; Harmon BG et al.; Avian peritoneal exudate macrophages, when exposed to phagocytic stimuli, produced an appreciable oxidative burst as measured by production of chemiluminescence, superoxide anion, and hydrogen peroxide . Metabolic inhibitors of the oxidative burst and scavengers of oxygen radicals clearly inhibited macrophage chemiluminescence, but had no significant effect on macrophage bactericidal activity against Escherichia coli or fungistatic activity against Candida tropicalis . Therefore, avian macrophages were capable of oxygen-independent bactericidal and fungistatic activities.

NCI Monogr, 1990, (9), 43 - 5
Role of surveillance cultures in prevention and treatment of fungal infections; Walsh TJ; Fungal surveillance cultures have been studied as potential predictors of invasive or disseminated mycoses . Several studies have demonstrated that the presence of Candida tropicalis in mucosal surveillance cultures has a high predictive value for invasive fungal infection due to this pathogen in granulocytopenic patients . By comparison, surveillance cultures for Candida albicans have a poor positive predictive value for invasive fungal infection . The value of routine surveillance cultures of the nares for Aspergillus spp . has not been consistently confirmed . The use of surveillance cultures for less common fungal pathogens, such as Trichosporon beigelii, also remains unclear . Fungal surveillance cultures of the inanimate hospital environment have proven useful in identifying the source of conidia in well-defined clusters or outbreaks of nosocomial aspergillosis and other mycoses . As investigational tools, fungal surveillance cultures also may be useful for studying the effects of new antifungal agents on mucosal flora . Fungal surveillance cultures, especially for C . tropicalis and possibly Aspergillus spp . in high-risk populations, may be useful when a pathogen-directed approach to antifungal therapy is used . However, the time required, diagnostic limitations, and expense of routine mucosal fungal surveillance cultures must be balanced against the effect of this information on therapeutic decisions . Empirical antifungal therapy and early diagnostic approaches for high-risk patients may obviate the need for routine fungal surveillance cultures while decreasing the frequency of invasive mycoses.

Chemotherapy, 1990, 36(6), 396 - 402
In vitro susceptibility testing of Candida isolates from clinical specimens to four antifungal agents; Ohkawa M et al.; A total of 231 isolates of Candida species including 163 of Candida albicans (132 of serotype A and 31 of serotype B), 42 of Candida tropicalis, 17 of Candida glabrata, 7 of Candida parapsilosis and 2 of Candida krusei were collected from clinical specimens in two medical centers, one in Japan and the other in the United States . The in vitro antifungal activities of amphotericin B, 5-fluorocytosine, miconazole and fluconazole were evaluated for the above isolates by means of a photo-read broth microdilution method for determination of the 30% inhibitory concentration (IC30) . The results showed that IC30 values of fluconazole were generally great compared with those of the other drugs, regardless of Candida species . The isolates of C . albicans serotype B showed a significantly lessened susceptibility to 5-fluorocytosine compared with those of serotype A (p less than 0.01) . This method has the advantage of objective measurement, thus minimizing observer variability as compared with the agar dilution test.

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