J Med Vet Mycol, 1990, 28(4), 267 - 73 Cell-constituent polyamines in Candida species and new biotyping of Candida albicans, Candida tropicalis and Candida parapsilosis strains; Bezjak V et al.; Intracellular amines from three Candida species were extracted and chemically derivatized by a modification of the Seiler procedure . The results of qualitative determinations of 13 amines in 20 strains each of Candida albicans, Candida tropicalis and Candida parapsilosis are presented . This report is the first to describe the detection of amines other than the classical spermine, spermidine, putrescine and cadaverine in yeasts and fungi . Characteristic profiles due to the presence or absence of particular amines in the Candida species studied are demonstrated . Although these could not be used as strict differential markers at the species level, biotyping schemes based on amines are proposed to differentiate strains of C . albicans, C . tropicalis and C . parapsilosis. J Med Vet Mycol, 1990, 28(1), 3 - 14 Demonstration of fungal proteinase during phagocytosis of Candida albicans and Candida tropicalis; Borg M et al.; The extracellular acid proteinase of Candida albicans and Candida tropicalis was monitored in vitro during phagocytosis by murine peritoneal macrophages . Fungal blastospores were quickly ingested by the thioglycolate-elicited macrophages and the intracellular blastospores partly resisted killing and started to grow out after 6 h incubation, causing destruction of the macrophage . Proteinase antigen appeared on fungal cells after 30 min in culture medium containing 10% fetal calf serum . The antigen was detected on ingested blastoconidia and filamentous cells of C . albicans serotype A . The proteinase antigen was also expressed by blastoconidia of C . albicans serotype B but was missing on the filamentous cells of this serotype . Isolates of C . tropicalis behaved similarly to C . albicans serotype A . The acid proteolytic activity of Candida cells was confirmed by the haemoglobin test on culture supernatants . Lysates of infected and noninfected phagocytes showed a differential acid proteolytic activity; noninfected macrophages revealed rising activity, while infected macrophages showed a distinct reduction of activity . The proteolytic activity of lysates of noninfected cells is due to lysosomal cathepsin-D . Cathepsin-D was also most likely to be responsible for the declining proteolytic activity in lysates from infected phagocytes; such lysates contained increasing amounts of fungal proteinase antigen . The differential kinetics of this antigen and the total acid proteolytic activity in the lysates suggest a conflict between microbial and lysosomal hydrolases in infected phagocytes . The outcome of this conflict depends on the number and hydrolytic activity of the ingested yeasts and may be decisive in the progress of infection. J Hyg Epidemiol Microbiol Immunol, 1990, 34(2), 129 - 34 The in vitro susceptibility of some mycotic agents to a new orally active triazole, itraconazole; Otcenasek M; The growth-inhibitory effect of itraconazole against 85 strains of mycotic agents belonging to 37 species was studied . MIC values were determined in a microtiter broth dilution method using casitone medium . The widest range of susceptibility was noted for yeasts . More strains of Candida albicans, Candida, spp . and Torulopsis spp . were found to be highly sensitive (MIC = 0.22 mg l-1), a single strain of Candida tropicalis and Rhodotorula glutinis was fully resistant (MIC = 200 mg l-1) . Most isolates of yeasts were susceptible at concentrations of 0.19 to 0.78 mg l-1 . Itraconazole showed the best activity against the clinical isolates of Aspergillus . Of the 16 strains tested, 12 isolates had MIC values less than or equal to 0.09 mg l-1 . The drug was inhibitory for the agents of adiaspiromycosis at a narrow range of concentrations (0.19-0.39 mg l-1) . greater variation in the response was noted for Geotrichum candidum (0.045-3.12 mg l-1) . Zygomycetous fungi (Mucor, Rhizopus) were mostly highly resistant (100-200 mg l-1) . Very susceptible strains of Absidia spp . were an exception . In most genera and species, the results of this study were in agreement with the previously reported data of foreign authors. Ann Pharm Fr, 1990, 48(1), 17 - 22 {Research of antifungal substances secreted by higher fungi in culture}; Pujol V et al.; A screening in vitro of antifungic activity of 24 strains of Basidiomycetes was realized with their culture filtrate . Lycoperdon perlatum Pers . = Pers., Oudemansiella platyphylla (Pers . ex Fr.) Mos., Agrocybe dura (Bolt) Singer have shown an activity against Candida albicans, Candida tropicalis and Aspergillus fumigatus; Pholiota spumosa (Fr.) Singer towards Botrytis cinerea and Lycoperdon perlatum Pers . = Pers . towards Alternaria solani, Botrytis cinerea and Verticillium dahliae . More extensive studies of the kinetics of antifungic substances production from Lycoperdon perlatum Pers . = Pers . were effectued with 2 different media and 2 different seeding technics on the strains which were sensible to the inhibitory character of this Gasteromycete. Biochemistry, 1989 Dec 12, 28(25), 9633 - 40 Mechanism and inhibition of delta 24-sterol methyltransferase from Candida albicans and Candida tropicalis; Ator MA et al.; The S-adenosyl-L-methionine: delta 24-sterol methyltransferase from Candida albicans has been solubilized with a mixture of octyl glucoside and sodium taurodeoxycholate . The enzyme has an apparent molecular weight of approximately 150,000 as measured by gel filtration chromatography . Zymosterol is the preferred substrate for the microsomal methyltransferase . Other nuclear double bond isomers support reduced rates of methenylation, while sterols which bear methyl groups at C-4 or C-14 are not substrates . Initial velocity and product inhibition studies are consistent with a rapid equilibrium ordered kinetic mechanism . A series of novel sterol analogues which contain heteroatoms substituted for C-24 or C-25 have been kinetically characterized as dead-end inhibitors of the methyltransferase, revealing three distinct mechanisms of interaction with the enzyme . Sterols which contain positively charged moieties in these positions are particularly potent inhibitors, supporting the proposed intermediacy of C-24 and C-25 carbocations . The methyltransferase is reversibly inhibited by low concentrations of 24-thiasterols, while behavior consistent with mechanism-based enzyme inactivation is apparent at higher concentrations . Possible mechanisms for this novel inactivation reaction are discussed. J Clin Microbiol, 1989 Nov, 27(11), 2426 - 8 Single-source outbreak of Candida tropicalis complicating coronary bypass surgery; Isenberg HD et al.; Candida tropicalis was isolated from the sternal wounds of eight coronary bypass patients from 18 to 89 days postoperatively; infections were limited to soft tissue in five patients but involved the sternum in three patients . Analysis of surgery records implicated one individual as the potential source of the yeast; this was confirmed by microbiological studies of fingertips and nasopharynx cultures of all personnel in contact with these patients . Only the suspect nurse, then acting as a scrub nurse and not as a circulator, infected the eight patients . Her removal from the cardiac team terminated the cluster outbreak. Diagn Microbiol Infect Dis, 1989 Nov-Dec, 12(6), 521 - 3 Rapid identification of Candida albicans using 4-methylumbelliferyl N-acetyl-beta-galactosaminide; Dalton MT et al.; Rapid identification of Candida albicans is performed mainly by the germ-tube test . However, recent reports have suggested that up to 5% of C . albicans species can give false negative results . We describe the use of 4-Methylumbelliferyl N-acetyl-beta-D-galactosaminide (4-MAG) conjugate as an alternative to the germ-tube test . Our results indicate that, in comparison to the germ-tube test, the 4-MAG test has a sensitivity of 100% and a specificity of 92% . Candida tropicalis can give false-positive results, and that a further screening test is required to identify this species . Problems reading end-points were not encountered. Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1989 Nov, 22(4), 242 - 8 Production of dodecanedioic acid from n-dodecane by yeasts; Liu WH et al.; In order to develop a process for production of dodecanedioic acid (DC-12) as starting materials for the chemical industry, a n-dodecane assimilating and DC-12-producing yeast was isolated from Taiwan soil . The taxonomical characteristics of this newly isolated yeast were examined and it was identified as Candida tropicalis NTU-512 . The newly isolated strain was improved successively by NTG mutagenesis . A high potency DC-12-producing mutant 91 which showed slight growth both in DC-12 and n-dodecane enrichment media was isolated . The DC-12 productivity of mutant 91 reached 3,326 mg/I by shaking culture at 30 degrees C for 72 h . This figure is 2.5-fold that of the parent strain (1,310 mg/I). Mycoses, 1989 Nov, 32(11), 578 - 80 Fungi isolated from lymph nodes of buffaloes; Pal M et al.; A preliminary study was undertaken to investigate the occurrence of fungi in various lymph nodes of domestic buffaloes (Bubalus bubalis), subjected to slaughter for human consumption at a local abattoir . Fungi could be demonstrated in 6 out 42 specimens when examined by mycological techniques . Molds were represented by 5 isolates viz., Aspergillus fumigatus (2), A . flavus, A . niger and A . terreus . Candida tropicalis was the only yeast isolate recovered from the lymph node . The significance of fungi in relation to meat inspection is discussed. FEBS Lett, 1989 Oct 9, 256(1-2), 128 - 34 Heterogeneity within the alkane-inducible cytochrome P450 gene family of the yeast Candida tropicalis; Sanglard D et al.; The reexamination of a genomic lambda gt11 Candida tropicalis expression library for the presence of genes related to the previously reported alkane-inducible cytochrome P450alk gene (P450alk), which is the first member of the P450LII gene family, was undertaken . A positive clone with a DNA fragment having 69% similarity with a portion of P450alk was isolated . As in the case of P450alk, this new putative P450 gene was also induced by tetradecane when C . tropicalis was grown on this carbon source and was therefore named P450alk2, P450alk1 corresponding to the first isolated P450 gene . In addition to P450alk2, the existence of other P450alk-related genes is suggested by the hybridization pattern of P450alk1 and P450alk2 probes with the C . tropicalis genomic DNA . The P450LII gene family in C . tropicalis appears therefore to include several different members . This heterogeneity is presently a unique feature within yeast P450 gene families and resembles the situation existing in P450 gene families of higher eukaryotes. J Pediatr, 1989 Oct, 115(4), 561 - 7 Role of flexible bronchoscopy in the diagnosis of pulmonary infiltrates in pediatric patients with cancer; Stokes DC et al.; We reviewed 60 consecutive flexible bronchoscopies done during a 36-month period in 48 pediatric cancer patients with undiagnosed pulmonary infiltrates . Diagnostic procedures during bronchoscopy included 40 brushings, 50 bronchoalveolar lavages, and 6 transbronchial and mucosal biopsies . A total of 16 specific diagnoses were made by bronchoscopy (27% diagnostic yield), including infection (12), pulmonary leukemia (3), and lymphoma (1) . The largest proportion of specific diagnoses came from lavage (14/50) and the smallest from brushings (1/40) . Biopsies were also useful for selected patients . The low overall yield for bronchoscopy was probably due to the routine use of empiric broad-spectrum antibiotics and antifungal therapy, as well as trimethoprim-sulfamethoxazole prophylaxis for Pneumocystis carinii pneumonitis . Subsequent specific diagnoses were obtained by other procedures (open biopsy, needle aspiration, or autopsy) for 10 patients with negative bronchoscopy results and 3 patients with diagnostic bronchoscopies . These additional diagnoses included 7 infections (Pneumocystis carinii (1), Candida tropicalis (1), cytomegalovirus (1), and Aspergillus (4), and 6 other diagnoses with nonspecific histologic findings . A positive bronchoscopy result may be useful, but negative bronchoscopy findings do not justify delaying other diagnostic procedures or discontinuing antibiotic and antifungal therapy in children with cancer and pulmonary infiltrates. Mycopathologia, 1989 Oct, 108(1), 11 - 9 Immunosuppression in experimental cryptococcosis in rats: modification of macrophage functions by T suppressor cells . Macrophages functions in cryptococcosis; Rubinstein HR et al.; The influence of lymphocytes on the modulation of macrophage functions in altered immune states induced by Cryptococcus neoformans infection in rats has been investigated . In this report we observed a decrease of 'in vitro' phagocytic activity by peritoneal cells (PC) from rats that received T suppressor cells induced by cryptococcal infection, against both the same microorganism that stimulated this suppressor population (p less than 0.05) and another non-pathogenic primary yeast (Candida tropicalis), (p less than 0.02) . The microbicide function of the PC from these animals present a significant decrease in challenge by C . tropicalis (p less than 0.002) when compared with PC from animals transferred with T normal cells . The transference of T suppressor cells induced by cryptococcal infection in animals immunized with human serum albumin-complete Freund's adjuvant (HSA-CFA) produces a significant alteration of the phagocytosis to HSA-human red cells (HSA-HRC) when compared with the phagocytosis observed in animals that received T normal cells or the phagocytosis of normal animals (p less than 0.001) . We could also observe that the DTH to HSA studied during 30 days was negative in rats transferred with PC sensitizated with HSA and treated with suppressor T cells, when compared with the DTH response of animals transferred with PC-HSA cocultured with normal cells (p less than 0.05 21st day) . The data presented in this paper illustrated that following infection of rats with C . neoformans there is a change in some population of accessory cells behavior reflected by the modification of several functions, such as phagocytosis, lytic activity and antigen presentation. J Biochem (Tokyo), 1989 Sep, 106(3), 474 - 8 Peroxisomal acetoacetyl-CoA thiolase and 3-ketoacyl-CoA thiolase from an n-alkane-utilizing yeast, Candida tropicalis: purification and characterization; Kurihara T et al.; Acetoacetyl-CoA thiolase (Thiolase I) and 3-ketoacyl-CoA thiolase (Thiolase III) found in peroxisomes of an n-alkane-utilizing yeast, Candida tropicalis pK 233, were each purified to homogeneity by successive column chromatographies . Thiolase I was composed of six identical subunits whose molecular masses were 41,000 Da, and Thiolase III was a homodimer composed of 43,000 Da subunits . The results of limited proteolysis of the respective thiolases indicated that they were quite different in peptide components . Furthermore, these enzymes were immunochemically distinguishable . The kinetic studies showed that the substrates with long chains were degraded exclusively by Thiolase III, while acetoacetyl-CoA was degraded preferentially by Thiolase I . Thus, in the yeast, the complete degradation of fatty acids is suggested to be carried out efficiently in peroxisomes. Antimicrob Agents Chemother, 1989 Sep, 33(9), 1443 - 6 Comparison of in vivo activity of fluconazole with that of amphotericin B against Candida tropicalis, Candida glabrata, and Candida krusei; Fisher MA et al.; Fluconazole (UK-49,858) is a new oral bis-triazole antifungal agent with demonstrated activity against Candida albicans . Because of the increasing importance of infections due to other species of Candida, we studied the efficacy of fluconazole in a rat model of established systemic candidiasis, using clinical isolates of C . tropicalis, C . glabrata, and C . Krusei . In normal rats, oral fluconazole at both 20 and 80 mg/kg per day for 7 days reduced both kidney and liver titers of C . tropicalis and C . glabrata compared with those in control animals and was only slightly inferior to amphotericin B . Both fluconazole and amphotericin B were ineffective in reducing kidney titers of C . krusei, but amphotericin B was more effective than fluconazole in reducing liver titers . Fluconazole showed no increased efficacy at the higher dose of 80 mg/kg per day compared with 20 mg/kg per day in any experiment . These results suggest that oral fluconazole may be useful in the treatment of established disseminated candidiasis caused by species other than C . albicans . Further in vivo studies are needed, however, to define minimum effective doses and length of therapy and to test additional Candida isolates. Antimicrob Agents Chemother, 1989 Aug, 33(8), 1391 - 2 Evaluation of cilofungin, a lipopeptide antifungal agent, in vitro against fungi isolated from clinical specimens; Hanson LH et al.; Cilofungin (LY121019) is a new lipopeptide antifungal drug . We tested this drug against 141 pathogenic fungal isolates . All fungal species were tested by broth dilution at 35 degrees C . Malassezia furfur was tested by agar dilution . The results demonstrate the specificity of cilofungin activity . Candida albicans, Candida tropicalis, and Malassezia pachydermatis were highly susceptible, whereas Candida parapsilosis, Candida pseudotropicalis, Candida krusei, Torulopsis glabrata, Blastomyces dermatitidis, Cryptococcus neoformans, Aspergillus species, M . furfur, and Paracoccidioides brasiliensis were more resistant. Appl Environ Microbiol, 1989 Aug, 55(8), 1974 - 80 31P nuclear magnetic resonance study of the effect of azide on xylose fermentation by Candida tropicalis; Lohmeier-Vogel E et al.; Maximal ethanol production by Candida tropicalis grown on xylose was obtained at an oxygen transfer rate of 5 to 7 mmol/liter per h . Addition of 0.2 mM azide increased the ethanol yield by a factor of 3 to 4, based on the cell mass produced, and decreased the formation of the by-product xylitol by 80% . In the presence of azide, ethanol was reassimilated before the carbon source was depleted . At all oxygenation levels studied, azide caused 25 to 60% of the carbon to be lost, most probably as carbon dioxide . Identical spectra were obtained with 31P nuclear magnetic resonance spectroscopy performed on extracts of C . tropicalis grown on xylose in the absence and presence of azide . Azide lowered the levels of sugar phosphates . Enzymatic analysis showed extremely low levels of fructose 1,6-diphosphate compared with the levels obtained in the absence of azide, while the level of malate, a citric acid cycle intermediate, was not influenced by azide . 31P nuclear magnetic resonance spectroscopy performed on xylose-grown whole cells of C . tropicalis showed that azide lowered the intracellular pH, inhibited the uptake of external Pi, and decreased the buildup of polyphosphate in relation to results with untreated cells . Similar results were obtained with the uncoupler of oxidative phosphorylation carbonyl cyanide m-chlorophenylhydrazone (CCCP), except that CCCP treatment led to extremely high levels of internal Pi . The dual effect of azide as a respiratory inhibitor and as an uncoupler is discussed with respect to the metabolism and product formation in xylose-assimilating C . tropicalis. Biochem Biophys Res Commun, 1989 Jul 31, 162(2), 646 - 50 Accumulation of cAMP in the cells of Candida tropicalis at an early stage of ethanol-induced filamentous growth and its prevention by myo-inositol; Omi K et al.; Phosphatidylinositol metabolism is enhanced in the cells of Candida tropicalis Pk 233 at an early stage of filamentous growth caused by ethanol, and myo-inositol prevents the ethanol-induced changes in the metabolism and morphology {Uejima et al . (1987) FEBS Lett . 214, 127-129} . The accumulation of cAMP and an increase in adenylate cyclase activity were observed in the cells grown with ethanol to the mid-log phase . Myo-inositol abolished these effects of ethanol also . The activity of cAMP phosphodiesterase was affected by neither ethanol nor myo-inositol . These results suggest that the inositol phospholipid-linked and cAMP-linked signaling pathways may be involved in the mechanism of ethanol-induced filamentous growth of this yeast and also that myo-inositol would affect morphogenesis by controlling these pathways. Am J Perinatol, 1989 Jul, 6(3), 347 - 8 Candida tropicalis empyema associated with acquired gastropleural fistula in a newborn infant; Malik S et al.; A case of congenital left-sided diaphragmatic hernia complicated by formation of gastropleural fistula and Candida tropicalis empyema is presented . The possibility of pleural fistula should be considered in any undiagnosed case of pleural effusion, pneumothorax, empyema, hemothorax, or hydropneumothorax. Mikrobiyol Bul, 1989 Jul, 23(3), 210 - 9 {The production of ethyl alcohol in waste sulfite liquor by free and immobilized Candida tropicalis ceppo 571 using batch and continuous systems}; Guven M et al.; In this study, the free and immobilized cells of Candida tropicalis ceppo 571 yeast were used for the production of ethanol . The yield of ethanol under anaerobic conditions was investigated . Batch technique in flasks and continuous in tubular bioreactor were also applied and the results were compared . It has been shown that the production of ethanol by the free cells was 2.36 times higher than the one by the immobilized cells . The continuous process resulted as the free cells produced ethanol 1.25 times more than the immobilized cells did. Biochem Biophys Res Commun, 1989 Jun 15, 161(2), 843 - 50 Molecular cloning and characterization of the primary structure of the alkane hydroxylating cytochrome P-450 from the yeast Candida maltosa; Schunck WH et al.; A cDNA library was established starting from poly(A) RNA of n-alkane-grown Candida maltosa cells and cDNA clones were isolated containing the entire coding sequence for the alkane hydroxylating cytochrome P-450 . The deduced protein consists of 521 amino acids, contains two putative transmembrane segments in the N-terminal region and has a characteristic heme-binding sequence in the C-terminal part . Sequence alignments with members of 11 reported cytochrome P-450 families revealed a strong homology to an alkane-inducible cytochrome P-450 from Candida tropicalis. J Bacteriol, 1989 Jun, 171(6), 3586 - 9 Interspecific complementation analysis by protoplast fusion of Candida tropicalis and Candida albicans adenine auxotrophs; Corner BE et al.; A protocol employing inositol starvation was used to isolate proline and adenine auxotrophs of Candida tropicalis . Interspecific hybrids between red adenine auxotrophs of C . tropicalis and Candida albicans were formed by protoplast fusion . These C . tropicalis red adenine auxotrophs were shown to fall into two complementation groups by crossing them with a known C . albicans ade1 tester strain . It is suggested that these two groups correspond to the ade1 and ade2 mutants of Saccharomyces cerevisiae and C . albicans and that these defined mutants may be useful in attempts to develop transformation systems for C . tropicalis. Infect Control Hosp Epidemiol, 1989 Jun, 10(6), 280 - 3 Candida tropicalis; Gelfand MS; C tropicalis is a frequent and virulent pathogen in neutropenic patients . Infection control personnel should be familiar with this microorganism and the significance of a positive culture for C tropicalis in a setting of poor host resistance. J Gen Microbiol, 1989 Jun, 135 ( Pt 6), 1423 - 30 Ammonium assimilation by Candida albicans and other yeasts: evidence for activity of glutamate synthase; Holmes AR et al.; Activities and properties of the ammonium assimilation enzymes NADP+-dependent glutamate dehydrogenase (GDH), glutamate synthase (GOGAT) and glutamine synthetase (GS) were determined in batch and continuous cultures of Candida albicans . NADP+-dependent GDH activity showed allosteric kinetics, with an S0.5 for 2-oxoglutarate of 7.5 mM and an apparent Km for ammonium of 5.0 mM . GOGAT activity was affected by the buffer used for extraction and assay, but in phosphate buffer, kinetics were hyperbolic, yielding Km values for glutamine of 750 microM and for 2-oxoglutarate of 65 microM . The enzymes GOGAT and NADP+-dependent GDH were also assayed in batch cultures of Saccharomyces cerevisiae and three other pathogenic Candida spp.: Candida tropicalis, Candida pseudotropicalis and Candida parapsilosis . Evidence is presented that GS/GOGAT is a major pathway for ammonium assimilation in Candida albicans and that this pathway is also significant in other Candida species. Eur J Clin Microbiol Infect Dis, 1989 Jun, 8(6), 564 - 7 Fungicidal activity of cilofungin (LY121019) alone and in combination with anticapsin or other antifungal agents; Pfaller M et al.; Cilofungin (LY121019) was shown to have potent fungicidal activity against clinical isolates of Candida albicans and Candida tropicalis but not Candida parapsilosis . Fungicidal activity was evident against both replicating and non-replicating Candida albicans and was progressive over the first 12 h of incubation . The combination of cilofungin (LY121019) with anticapsin but not with amphotericin B, ketoconazole or 5-fluorocytosine resulted in synergistic fungicidal activity . This compound warrants further investigation of its safety and efficacy in the treatment of Candida infections. Biochem Biophys Res Commun, 1989 May 15, 160(3), 1257 - 66 Disruption of the Saccharomyces cerevisiae gene for NADPH-cytochrome P450 reductase causes increased sensitivity to ketoconazole; Sutter TR et al.; Strains of Saccharomyces cerevisiae deleted in the NADPH-cytochrome P450 reductase gene by transplacement are 200-fold more sensitive to ketoconazole, an inhibitor of the cytochrome P450 lanosterol 14 alpha-demethylase . Resistance is restored through complementation by the plasmid-borne wild type gene from either S . cerevisiae or Candida tropicalis . Neither Southern hybridization nor Western immunoblot techniques provided evidence for a second NADPH-cytochrome P450 reductase gene, suggesting that an alternate pathway may provide for the functions of this reductase in S . cerevisiae. Rev Infect Dis, 1989 May-Jun, 11(3), 379 - 90 Fungemia caused by Candida species and Torulopsis glabrata in the hospitalized patient: frequency, characteristics, and evaluation of factors influencing outcome; Komshian SV et al.; We reviewed 135 cases of candidemia occurring between 1983 and 1986 to examine oncologic and nononcologic populations and assess factors for survival . Candida albicans was the most common species (51%); Candida tropicalis occurred most frequently in leukemia patients (57%), whereas Candida parapsilosis and Torulopsis glabrata were associated with solid tumors and nononcologic diseases . Risk factors identified were: preceding surgery, antibiotics, cannulas, and steroids in solid tumor and nononcologic diseases; and chemotherapy and neutropenia with hematologic malignancies . Even transient cannula-associated candidemia was not a benign process . Intravenous cannulas were common portals of entry (39%) in debilitated patients without cancer (59%) and were associated with high mortality (55%) . Overall mortality was 59%, candidemia directly contributing to death in 75% of cases . In patients with candidemia, failure to initiate therapy with amphotericin B had a negative influence on outcome, whereas analysis of the entire group identified severity of underlying illness as the dominant cofactor influencing outcome. Med Clin (Barc), 1989 Apr 8, 92(13), 503 - 6 {Post-transfusion graft versus host disease in a patient with Hodgkin's disease}; Tomas Martinez JF et al.; A female with Hodgkin's disease developed graft versus host disease (GVHD) after the administration of two units of packed red cells . Ten days after the transfusion she developed fever and rash, with subsequent hepatic and intestinal disease and a profound bone marrow aplasia . She died from a Candida tropicalis sepsis . The diagnosis of GVHD was made on the basis of clinical and histological criteria . We review this uncommon complication of hemotherapy, with special emphasis on its differential clinical features and its prevention. Yeast, 1989 Apr, 5 Spec No, S437 - 9 Increase in cyclic AMP content with enhanced phosphatidylinositol turnover in the cells of Candida tropicalis during mycelial growth caused by ethanol; Kamihara T et al.; Ethanol causes mycelial growth of Candida tropicalis Pk 233, which is associated with enhanced metabolism of phosphatidylinositol at the mid-log phase of growth, and the effects of ethanol are prevented by concomitant addition of myo-inositol (FEBS Lett . 214, 127-129, 1987) . Ethanol induced also a marked increase in cellular content of cAMP at the mid-log phase, and myo-inositol abolished this effect of ethanol . The elevated level of cAMP content caused by ethanol was gradually lowered through the late-log and stationary phases and reached to control level . Very similar effects of ethanol and myo-inositol were observed in adenylate cyclase activity, while the activity of cAMP phosphodiesterase was not affected by ethanol . The ethanol-induced change in cAMP content was therefore ascribed to that in adenylate cyclase activity . These results suggested that cAMP plays an important role in combination with phosphatidylinositol turnover in the development of mycelial form in this dimorphic yeast. Gene, 1989 Mar 15, 76(1), 121 - 36 Characterization of the alkane-inducible cytochrome P450 (P450alk) gene from the yeast Candida tropicalis: identification of a new P450 gene family; Sanglard D et al.; The P450alk gene, which is inducible by the assimilation of alkane in Candida tropicalis, was sequenced and characterized . Structural features described in promoter and terminator regions of Saccharomyces yeast genes are present in the P450alk gene and some particular structures are discussed for their possible role in the inducibility of this gene . Expression of the P450alk gene was achieved in Saccharomyces cerevisiae using the yeast alcohol dehydrogenase expression system after removal of the P450alk gene flanking regions . The resultant expressed protein had a molecular mass slightly greater than that of P450alk from C . tropicalis . This alteration did not prevent the function and the localization of P450alk expressed in S . cerevisiae, as this organism showed an acquired microsome-bound activity for the terminal hydroxylation of lauric acid . The deduced P450alk amino acid sequence was compared with members of the nine known P450 gene families . These comparisons indicated that P450alk had a low relationship with these members and was therefore the first member (A1) of a new P450 gene family (LII). Mycoses, 1989 Mar, 32(3), 145 - 50 Comparative activity in vitro of cilofungin (LY 121019) with other agents used for treatment of deep-seated Candida infections; Rennie RP et al.; Cilofungin (LY 121019) is a semi-synthetic lipopeptide antifungal agent that inhibits beta-1,3-D-glucan synthase activity in the cell wall of yeasts . Clinical strains of Candida species were tested for susceptibility to cilofungin and seven other antifungal agents . Candida albicans and Candida tropicalis were susceptible to cilofungin with mean MICs of 0.3 and 0.08 microgram/ml respectively . For most other species the mean MICs were greater than 1-2 micrograms/ml of cilofungin . Studies on the paradoxical growth effect observed with cilofungin in Sabouraud broth showed that, at high concentrations of cilofungin, sufficient damage occurred to make damaged cells highly susceptible to killing by fresh cilofungin . The damaged cells also had increased susceptibility to other antifungal agents to which they were normally resistant . These observations indicate that cilofungin may be a useful agent for the treatment of many invasive Candida infections, either alone or in combination with certain other antifungal agents. J Clin Microbiol, 1989 Mar, 27(3), 400 - 4 Protein and enzyme electrophoresis profiles of selected Candida species; Lehmann PF et al.; The cellular protein profiles and malate dehydrogenases, superoxide dismutases, alkaline phosphatases, and esterases from whole cell extracts of Candida spp . were studied with polyacrylamide gel electrophoresis . We investigated isolates that differed in their ability to assimilate sucrose as the sole carbon source . The protein and enzyme patterns of Candida tropicalis and its sucrose-negative variant "Candida paratropicalis Baker, Salkin, Pincus et D'Amato" were indistinguishable . Although the cellular protein and superoxide dismutase patterns of Candida albicans and its sucrose-negative variant "Candida stellatoidea" were quite similar, differences were noted in the profiles of the other enzymes studied . In addition, the C . stellatoidea isolates were found to be separable, on the basis of their enzyme profiles, into the same two types that have been reported by Kwon-Chung et al . (K.J . Kwon-Chung, B.L . Wickes, and W.G . Merz, Infect . Immun . 56:1814-1819, 1988). Indian J Exp Biol, 1989 Mar, 27(3), 224 - 6 Hydrocarbon emulsification by Candida tropicalis and Debaryomyces polymorphus; Singh M et al.; Potentiality of C . tropicalis and D . polymorphus, to produce surface active compounds (bioemulsifiers/biosurfactants) during shake cultivation on hexadecane and oily waste was studied . Better emulsification activity, specific towards aromatic hydrocarbons, was observed with C . tropicalis culture broth . Emulsification activity of culture broth was quite stable and was unaffected by change in pH and by increasing the concentration of NaCl up to 5% . The activity was marginally affected by heating in boiling water bath for 15 min, but inhibited to the tune of 90% by 0.3% CaCl2 . The isolated bioemulsifying factor contained 40, 22 and 17.5% lipid, protein and carbohydrate, respectively. Infection, 1989 Mar-Apr, 17(2), 111 - 7 Secondary mycosis in surgery: treatment with fluconazole; Kujath P et al.; Together with the severity of the underlying disease, mycotic infections are assuming increasing significance in surgical patients under intensive care . 26 patients with severe internal mycotic infections were treated with fluconazole in an open clinical trial . The pathogens isolated were Candida albicans in 22 cases, Candida tropicalis in four, Torulopsis glabrata in three, Candida parapsilosis in two and Aspergillus fumigatus in one . Mixed infections were found in five cases . Most of this series of surgical patients had a severe, life-threatening post-operative condition . Their mean classification by the Apache score was 15.35 . The most frequent risk factors were previous injuries to intraabdominal hollow organs and extensive antibiotic therapy; and the peritoneum was, therefore, the commonest site of infection . Antimycotic therapy was with fluconazole at a dose of 200-400 mg daily for at least ten days . The mycosis was cured in 22 of the 26 patients, one of these being a case of severe mycotic peritonitis . Nine patients died of the underlying disease within four weeks of the beginning of treatment . Fluconazole was effective and well tolerated in the treatment of severe life-threatening infections in surgical patients. Recenti Prog Med, 1989 Mar, 80(3), 119 - 22 {Variability in immunologic aspects of Whipple's disease . Is a unitary interpretation possible?}; Sardeo G et al.; A case of Whipple's disease with histological and ultrastructural studies, characterized by unusual bacteriological and immunologic findings, is reported . Alpha hemolytic Streptococcus and Candida tropicalis were isolated from the culture of the intestinal biopsy specimens . The immunological function study showed a global defect both of humoral and cellular immunity . On the basis of the literature review, the Authors debate a unitary interpretation of the various immunological dysfunctions reported in this disease. South Med J, 1989 Feb, 82(2), 270 - 3 Intervertebral diskitis due to Candida tropicalis; Herzog W et al.; This report describes a case of Candida tropicalis intervertebral diskitis successfully treated with a brief course of amphotericin B followed by a longer course of ketoconazole . Candida tropicalis is an increasingly frequent pathogen in immunocompromised patients, and infection can become manifest weeks or months after an episode of neutropenia has resolved . The excellent response we observed in this patient adds to a growing body of clinical experience testifying to the effectiveness of ketoconazole in treating certain deep-seated candidal infection. Gene, 1989 Jan 30, 75(1), 119 - 26 The nucleotide sequence of POX18, a gene encoding a small oleate-inducible peroxisomal protein from Candida tropicalis; Szabo LJ et al.; We report the molecular cloning and nucleotide sequence of the nuclear gene, POX18, encoding an oleate-inducible peroxisomal protein from the yeast Candida tropicalis . POX18 has a single open reading frame of 381 nucleotides (nt), which encodes a protein of 127 amino acids . The predicted Mr of this protein is 13,792 . Codon usage in the expression of POX18 is non-random, and shows a pattern similar to that used for other peroxisomal genes from C . tropicalis and highly expressed genes from Saccharomyces cerevisiae . Northern analysis of total RNA from oleate-grown cells determined that POX18 mRNA is approximately 750 nt in length . The POX18 gene was expressed in vitro, which resulted in a single translation product that co-migrated in denaturing polyacrylamide gels with an abundant peroxisomal protein (apparent mass of 16 kDa) and was immunoprecipitated by an antiserum against peroxisomal protein. Rev Infect Dis, 1989 Jan-Feb, 11(1), 89 - 96 Periprosthetic candidal infections following arthroplasty; Darouiche RO et al.; Candidal infection after prosthetic arthroplasty has been reported in six previous cases, to which four cases are now added . Candida albicans was the offending organism in four patients, Candida parapsilosis in three, Candida tropicalis in two, and Candida (Torulopsis) glabrata in one . None of the 10 patients had evidence of disseminated candidiasis, and, except for the uniform presence of a prosthesis, other underlying factors that are generally associated with candidal infections were present in only three . Clinical features that distinguished periprosthetic from natural bone and joint infection included an older patient population, the usual lack of predisposing factors other than the prosthesis, and the absence of evidence of disseminated candidiasis . All patients were treated with removal of the prosthesis and antifungal therapy, consisting of amphotericin B alone (six patients) or combined with 5-fluorocytosine (three patients) or ketoconazole (one patient) . Infection appeared to be cured in nine of the 10 patients, but the follow-up was less than 1 year in five cases . Replacement with a new prosthetic joint was attempted in only two cases and successful in only one . Direct inoculation of organisms during surgery or transient unrecognized candidemia may initiate periprosthetic infection, which might then be promoted by favorable local factors, both mechanical and molecular . The role of prosthetic materials, candidal adhesins, and human factors such as fibronectin in initiating these infections has yet to be characterized. Avian Dis, 1989 Jan-Mar, 33(1), 177 - 81 In vitro microbicidal activity of avian peritoneal macrophages; Harmon BG et al.; Peritoneal exudate macrophages collected from 4-wk-old broilers were capable of in vitro bactericidal activity against Escherichia coli and fungistatic activity against Candida tropicalis . Bactericidal activity was inhibited by treatment of macrophages with 2-deoxyglucose . These assays may be used to evaluate macrophage function in normal, diseased, and immunomodulated birds or to assess the direct effects of cytokines and toxins on macrophage microbicidal activity. Jpn J Antibiot, 1989 Jan, 42(1), 31 - 9 {Clinical evaluation of fluconazole}; Oka S et al.; Clinical evaluation of fluconazole was performed on 12 cases of mycotic infections (7 cases of Candida esophagitis; one each case of cryptococcal meningitis with AIDS, Candida tropicalis fungemia and disseminated cryptococcosis in kidney transplant patient; 2 cases of Candida pneumonia) . Satisfactory responses were obtained except 1 case of Candida pneumonia in which clinical efficacy could not be evaluated . Hiccup was noted in 1 case during the fluconazole treatment . No other adverse reaction was observed . When 150 mg and 200 mg of fluconazole were administered orally to a patient with hemodialysis (HD) after HD on separate occasions, concentrations of the drug in serum at 20 hours after ingestion were 5.9 micrograms/ml and 11.6 micrograms/ml, respectively, and in cerebrospinal fluid (CSF) were 3.5 micrograms/ml and 9.2 micrograms/ml, respectively . Two clinical benefits were obtained in our studies . First, it was possible to treat the AIDS-patient as an outpatient with Candida esophagitis using orally administered fluconazole . Second, it was possible to treat the case of cryptococcal meningitis, in which relapse often occurs, to complete the therapy when the cryptococcal antigen in serum and CSF diminished to an undetectable level and to maintain the therapy preventing relapse without severe adverse effects . Ongoing and future clinical trials will define the specific roles of fluconazole more clearly in the treatment of systemic mycosis. Jpn J Antibiot, 1989 Jan, 42(1), 171 - 8 {Clinical efficacy of fluconazole in urinary tract fungal infections}; Nito H; Fluconazole, a new antifungal agent was administered to 7 patients with complicated urinary tract fungal infections . Patients were 6 males and 1 female with ages of 29 to 83 years, with underlying conditions of bladder tumor (3 patients), neurogenic bladder (3 patients) and hydronephrosis (1 patient) . Urinary fungi identified were Candida albicans in 5 patients and Candida tropicalis in 2 patients over 10(4) CFU/ml . These fungi were isolated at least twice in intervals of 5 to 7 days before treatment . Fluconazole was given either orally (4 patients) or intravenously (3 patients) in a dose of 50 mg per day . Clinical efficacies were excellent in 3 patients, moderate in 3 patients and poor in 1 patient, showing an efficacy rate of 85.7% . Mycologically, 6 Candida out of the 7 were eliminated . No side effects nor abnormal laboratory data were observed . In conclusion, fluconazole is effective and safe in the treatment of urinary tract fungal infections. Fukuoka Shika Daigaku Gakkai Zasshi, 1989, 16(3), 407 - 22 {Adherence of Candida albicans to acrylic surfaces}; Kaita H; The ability of Candida albicans IFO 1385 to adhere to acrylic and the partial characterization of an adhesive substance, named AS, which was isolated from the yeast, were studied in vitro . The results obtained were as follows: 1 . The cells cultured in the synthetic media (YNB) containing 500 mM galactose showed a much greater tendency to adhere than did those cells cultured in the YNB containing 500 mM glucose . 2 . More cells prepared by the standing cultivation adhered to acrylic than did those prepared by the stirring cultivation . 3 . A large number of the adherent cells was obtained when the acrylic plates were incubated at 37 degrees C for 90 min in the cell suspension at a concentration of 1.0 x 10(7) cells/ml . The plates were observed without staining . 4 . AS was isolated from the surface of C . albicans, grown on different carbon sources (50 mM glucose, 500 mM glucose and 500 mM galactose), by treatment with ultrasonication . 5 . Three different kinds of AS isolated from the three carbon sources were slightly soluble in distilled water . All were similar in composition to each other, and contained 62-68% carbohydrate (as glucose) and 23-26% protein (as BSA) . 6 . Silica particles adhered to acrylic coated with AS and pretreatment of acrylic with AS promoted C . albicans adhesion . However, similar pretreatment inhibited subsequent Candida glabrata and Candida krusei adhesion . As to subsequent adhesion of Candida tropicalis, no significant data were obtained . 7 . Adhesion assay using the silica particles, the adhesive ability of the AS was significantly reduced by treatment with trypsin or pronase E, but not with papain, alpha-amylase, dextranase or zymolyase. J Clin Microbiol, 1988 Nov, 26(11), 2307 - 12 Collaborative evaluation in seven laboratories of a standardized micromethod for yeast susceptibility testing; Guinet R et al.; The new micromethod for yeast susceptibility testing, MYCOTOTAL, was evaluated with 10 reference strains in seven laboratories . Ready-to-use microtitration plates and the same synthetic medium were used with two dilutions of imidazoles, flucytosine, and amphotericin B, permitting the categorization of each strain as susceptible, intermediate, or resistant . The results were compared with the MIC for each reference strain, and the repeatability and reproducibility were evaluated . The yeasts tested presenting different patterns of susceptibilities in reference MICs included six strains of Candida albicans, two strains of Candida tropicalis, one strain of Candida parapsilosis, and one strain of Torulopsis glabrata . For 4,200 antifungal agent-yeast results, the repeatability was 99.3% and the reproducibility was 96.3% . The correlation between the reference MICs and the category results was 91.5% for seven laboratories (and 92.7% for six laboratories excluding the laboratory which did not follow exactly the same protocol) . We observed only 7.9% minor discrepancies, 0.5% (0.29% for six laboratories) major discrepancies, and 0.1% uninterpretable results . The percentages of concording results were similar for each strain and each antifungal agent tested . The overall results indicated that MYCOTOTAL was a reliable and reproducible method, well correlated with reference MICs . This ready-to-use micromethod with the same medium for all antifungal agents would be an important step in the necessary standardization of yeast susceptibility testing. DNA, 1988 Nov, 7(9), 617 - 26 Primary structure of the cytochrome P450 lanosterol 14 alpha-demethylase gene from Candida tropicalis; Chen C et al.; We report the nucleotide sequence of the gene and flanking DNA for the cytochrome P450 lanosterol 14 alpha-demethylase (14DM) from the yeast Candida tropicalis ATCC750 . An open reading frame (ORF) of 528 codons encoding a 60.9-kD protein is identified . This ORF includes a characteristic heme-binding domain, HR2, common to all P450 proteins . This protein and the 14DM from Saccharomyces cerevisiae share 66.5% identical and 23.1% conservatively replaced amino acids in a 516-amino-acid alignment, and thus are orthologous forms of the P450LIA1 gene . Conversely, C . tropicalis 14DM shares relatively little sequence similarity with P450alk, the predominant P450 protein present when this organism is grown on n-alkanes . Sequence information of these three yeast P450s will be useful for structure-function analyses in the future. Ann Acad Med Singapore, 1988 Oct, 17(4), 551 - 3 Acute disseminated cutaneous candidiasis; Fong PH et al.; Acute disseminated candidiasis is a serious and difficult problem often seen in immunocompromised states . Appearance of a characteristic skin eruption is helpful in the diagnostic . We report below a case report of an eight year old girl with aplastic anemia who had received multiple courses of antibiotics . A profuse monomorphic papular nodular eruption subsequently appeared on the face, palms and soles . Candida tropicalis was identified from the skin biopsy taken from one such lesion. Gene, 1988 Sep 30, 69(2), 171 - 80 cDNA cloning and primary structure determination of the peroxisomal trifunctional enzyme hydratase-dehydrogenase-epimerase from the yeast Candida tropicalis pK233; Nuttley WM et al.; We report the isolation and nucleotide (nt) sequence determination of a cDNA encoding the peroxisomal trifunctional beta-oxidation enzyme hydratase-dehydrogenase-epimerase (HDE) from the yeast Candida tropicalis pK233 . Poly(A)+RNA isolated from C . tropicalis cells grown in oleic acid medium was used to construct a cDNA library in lambda gt11 . The library was screened with a polyclonal antiserum against HDE . A recombinant was confirmed to encode HDE by hybridization-selection translation and immunoprecipitation . The HDE cDNA (HDE) has a single open reading frame of 2718 nt, encoding a protein of 905 amino acids, not including the initiator methionine . The Mr of the protein is 99,350 . A partial gene duplication is believed to have occurred in the evolution of the HDE gene . Codon utilization in the gene is not random, with 86.0% of the amino acids specified by 23 preferentially used codons, a situation similar to that found in genes encoding peroxisomal catalase and the various fatty acyl-CoA oxidases from C . tropicalis . The increase in HDE activity in C . tropicalis cells grown in oleic acid medium as opposed to glucose medium is due, at least in part, to increased HDE-specific mRNA levels. Diagn Microbiol Infect Dis, 1988 Sep, 11(1), 1 - 9 Influence of in vitro susceptibility testing conditions on the anti-candidal activity of LY121019; Pfaller MA et al.; LY121019 is a new antifungal antimicrobic that is structurally similar to the lipopeptide agents echinocandin B and aculeacin A . Because of the importance of in vitro test conditions on the activity of other antifungal agents, we studied the effects of inoculum size, time and temperature of incubation, pH, and medium composition on the in vitro activity of LY121019 against Candida albicans, Candida tropicalis, and Candida parapsilosis . LY121019 was highly active against Candida albicans and Candida tropicalis and inactive against Candida parapsilosis . The in vitro activity of LY121019 is marked by a paradoxical dose-response with isolates of Candida albicans and Candida tropicalis and is influenced by choice of inoculum size, time and temperature of incubation, medium composition, and pH . We recommend the use of an inoculum size of less than 10(5) organisms/ml, a defined medium buffered to a pH of 7.0, and incubation at 30 degrees C for 24 hr for future in vitro studies of LY121019. Hinyokika Kiyo, 1988 Sep, 34(9), 1679 - 82 {Clinical efficacy of flucytosine on urinary candidiasis}; Yasumoto R et al.; An antifungal agent (Flucytosine) was used to treat urinary candidiasis in 9 patients who had an indwelling catheter and developed fungal colony counts greater than 10(4) . Among 9 patients with catheter drainage, urologic underlying diseases were benign prostatic hyperplasia in 7 and a neurogenic bladder in one patient all of whom had accompanied diabetes mellitus . Only one patient was supravesically diverted from the upper urinary tract through an indwelling catheter of bilateral ureterocutaneostomy after the removal of a tumorous bladder . All patients had previously received antimicrobials . Isolated strains of Candida were Candida albicans in 6, Candida tropicalis in 2, and Candida parapsilosis in one patient . Out of 9 patients having received daily administration of 1,500 mg Flucytosine for 2 weeks, 7 patients subsequently had no yield of fungal colony after the treatment . Minimum inhibitory concentration (MIC) of this agent was determined at the range of 0.1 to 0.2 microgram/ml in 5 patients with C . albicans and 0.2 microgram/ml in both patients with C . tropicalis . Otherwise, a high MIC of over 100 micrograms/ml indicating resistance to this agent was observed in only 2 patients with C . albicans and C . parapsilosis . Three of the 7 patients had recurrent urinary Candida infection even 2 weeks after the discontinuation of this antifungal therapy despite rapid and excellent eradication of urinary candidiasis . From these results, Flucytosine may be one of the most promising antifungal agent with a low MIC in the treatment of compromised urinary Candida infection and should be occasionally supplemented with a topical instillation of amphotericin B without any serious complication in the prevention of recurrence. Infect Immun, 1988 Sep, 56(9), 2495 - 8 Endothelial cell contraction increases Candida adherence to exposed extracellular matrix; Klotz SA et al.; Bovine vascular endothelial cells treated with EDTA, urea, or thrombin underwent a marked, reversible contraction resulting in exposure of the subendothelial extracellular matrix (ECM) . Candida yeasts adhered more to contracted monolayers than to confluent monolayers (P less than 0.01) by preferentially adhering to the ECM . Two strains of Candida albicans and one strain of Candida tropicalis bound avidly to exposed ECM, but Pseudomonas aeruginosa did not . However, treatment of endothelium with forskolin, which induces cell shape changes without exposure of the ECM, did not cause an increase in adherence. Antimicrob Agents Chemother, 1988 Sep, 32(9), 1331 - 5 Cilofungin (LY121019), an antifungal agent with specific activity against Candida albicans and Candida tropicalis; Hall GS et al.; Cilofungin (LY121019) is an antifungal agent that interferes with beta-glucan synthesis in the cells walls of fungi . The activity of this agent against 256 clinical isolates of yeasts was determined . It was found to be very active in vitro against Candida albicans (MIC for 90% of isolates {MIC90}, less than or equal to 0.31 microgram/ml; minimal fungicidal concentration for 90% of isolates {MFC90}, less than or equal to 0.31 micrograms/ml) and C . tropicalis (MIC90, less than or equal to 0.31 microgram/ml; MFC90, less than or equal to 0.31 microgram/ml) and moderately active against Torulopsis glabrata (MIC90 and MFC90, less than or equal to 20 micrograms/ml) . All C . parapsilosis, Cryptococcus, and Saccharomyces cerevisiae strains were resistant . The activity of cilofungin was affected by medium and inoculum size . Antibiotic medium no . 3 was used as the standard medium . Isolates of C . albicans and C . tropicalis demonstrated a paradoxical effect in Sabouraud dextrose broth and yeast nitrogen base broth in that growth was partially inhibited at MICs equivalent to those in antibiotic medium no . 3, but growth continued, in many instances, throughout all concentrations tested . There was decreased activity of cilofungin with inocula greater than 10(5) CFU/ml . The temperature and duration of incubation did not affect its activity. Eur J Biochem, 1988 Sep 1, 176(1), 159 - 63 Sequence of the Saccharomyces cerevisiae CTA1 gene and amino acid sequence of catalase A derived from it; Cohen G et al.; The nucleotide sequence of a 2785-base-pair stretch of DNA containing the Saccharomyces cerevisiae catalase A (CTA1) gene has been determined . This gene contains an uninterrupted open reading frame encoding a protein of 515 amino acids (relative molecular mass 58,490) . Catalase A, the peroxisomal catalase of S . cerevisiae was compared to the peroxisomal catalases from bovine liver and from Candida tropicalis and to the non-peroxisomal, presumably cytoplasmic, catalase T of S . cerevisiae . Whereas the peroxisomal catalases are almost colinear, three major insertions have to be introduced in the catalase T sequence to obtain an optimal fit with the other proteins . Catalase A is most closely related to the C . tropicalis enzyme . It is also more similar to the bovine liver catalase than to the second S . cerevisiae catalase . The differences between the two S . cerevisiae enzymes are most striking within four blocks of amino acids consisting of a total of 37 residues with high homology between the three peroxisomal, but low conservation between the S . cerevisiae catalases . The results obtained indicate that the peroxisomal catalases compared have very similar three-dimensional structures and might have similar targeting signals. Biochem J, 1988 Aug 1, 253(3), 845 - 9 Biosynthesis of soluble carnitine acetyltransferases from the yeast Candida tropicalis; Kozulic B et al.; Soluble carnitine acetyltransferase from Candida tropicalis is synthesized as a 76 kDa precursor, which is monomeric and possesses no or very little carnitine acetyltransferase activity . Maturation of the enzyme begins with proteolytic processing of the 76 kDa precursor to 64 and 57 kDa subunits . The processed subunits subsequently associate into two kinds of active oligomers; the 57 kDa subunits are assembled into a tetramer and the 64 kDa subunits into an octamer . Formation of these oligomers depends apparently on growth conditions, since both oligomers were present in cells grown in continuous culture, but cells grown batchwise contained only the tetrameric form of carnitine acetyltransferase. J Clin Microbiol, 1988 Aug, 26(8), 1448 - 59 Multiple Candida strains in the course of a single systemic infection; Soll DR et al.; Species and strain variabilities have been monitored during the history of a prolonged Candida infection in a single compromised bone marrow transplant patient by analyzing sugar assimilation patterns, high-frequency switching repertoires, and Southern blot hybridization patterns with two cloned mid-repeat sequences (Ca3 and Ca7) which are species specific for Candida albicans and one cloned mid-repeat sequence (Ct13-8) which is species specific for Candida tropicalis . Evidence is presented that during the course of this infection (i) two strains of C . albicans and three strains of C . tropicalis were distinguished by their switching repertoires, Southern blot hybridization patterns, and sugar assimilation patterns; (ii) the three C . tropicalis strains were in a high-frequency mode of switching; (iii) two C . tropicalis strains coexisted in the blood and three C . tropicalis strains coexisted in the throat at different times during the history of the infection; (iv) amphotericin B treatment selectively removed one of two C . tropicalis strains coexisting in the blood and this strain exhibited greater susceptibility to amphotericin B in vitro (the remaining strain was subsequently removed from the blood by flucytosine treatment); and (v) both the strain removed from the blood by amphotericin B and the strain removed from the blood by flucytosine reappeared several days later at another site of infection . It is demonstrated for the first time that C . tropicalis is capable of high-frequency switching of colony morphology just as C . albicans is, that there is more than one strain-specific switching repertoire in C . tropicalis, and that a C . tropicalis mid-repeat sequence can be used for discriminating species and assessing strain relatedness, as previously demonstrated for C . albicans mid-repeat sequences. J Clin Microbiol, 1988 Aug, 26(8), 1437 - 41 Multicenter evaluation of four methods of yeast inoculum preparation; Pfaller MA et al.; We initiated a comparative study of four methods of yeast inoculum preparation: a spectrophotometric method, the Wickerham card method, a hemacytometer method, and the Prompt inoculation system . The variability in inoculum size obtained when each method was applied to two strains each of Candida albicans, Candida tropicalis, Candida parapsilosis, Torulopsis glabrata, Cryptococcus neoformans, and Saccharomyces cerevisiae was analyzed in a single laboratory . Each method was performed in triplicate on the same day and on three separate days to provide estimates of within-day and between-day variations . Inoculum size was determined by viable colony counts . The greatest range of inoculum sizes was seen with the Wickerham card method . Viable counts ranged from 1.1 X 10(6) to 24.2 X 10(6) CFU/ml among the 12 yeast isolates . The greatest variation was observed with the Prompt system . Within-day coefficients of variation averaged 19% (range, 4 to 45%), and between-day coefficients of variation averaged 22% (range, 3 to 51%) . Variation between laboratories was evaluated by comparing inoculum values obtained by each method in three different laboratories for two strains of C . albicans . The spectrophotometric method was the least variable and the Wickerham card and hemacytometer methods were the most variable methods between laboratories . The spectrophotometric method is recommended as the method of choice for preparation of a standardized inoculum suspension for susceptibility testing of yeasts. J Infect Dis, 1988 Jul, 158(1), 80 - 8 Evaluation of single-drug and combination antifungal therapy in an experimental model of candidiasis in rabbits with prolonged neutropenia; Thaler M et al.; We developed an experimental model of candidiasis in rabbits with prolonged neutropenia . Rabbits were made neutropenic with cytosine arabinoside (Ara-C) administered through an indwelling silastic catheter that had been surgically implanted in the external jugular vein . Neutropenia was sustained with intravenous Ara-C, and bacterial complications were prevented with parenteral ceftazidime plus ampicillin . Candidiasis was established by intravenously administering Candida albicans or Candida tropicalis (1-2 x 10(5) colony-forming units) and resulted in hepatic and splenic lesions that mimicked those associated with hepatosplenic candidiasis in humans . The kidney proved to be the site most refractory to eradication of Candida spp . and offered a target organ for assessing antifungal therapy . We evaluated amphotericin B, 5-flucytosine, ketoconazole, and rifampin, alone and in combination . Although each agent reduced the colony counts of Candida in the liver, spleen, and lung, the combination of amphotericin B and 5-flucytosine was the only regimen effective in eradicating renal candidiasis. J Am Vet Med Assoc, 1988 Jun 1, 192(11), 1577 - 80 Nonsurgical management of ruptured urinary bladder in a critically ill foal; Lavoie JP et al.; A small tear in the urinary bladder of a severely debilitated 4-day-old foal was managed with an indwelling urinary catheter connected to a urine collecting system . Fluid therapy, parenteral nutrition, and antimicrobial agents were used during the initial management of the ruptured bladder . Aseptic technique for catheter care and systemic administration of antimicrobial agents prevented the development of bacterial cystitis . Catheter management required constant monitoring, but the bladder defect was sealed within 5 days . Fungal arthritis caused by Candida tropicalis, immune-mediated anemia, diarrhea, constipation, venous thrombosis, and pneumonia were observed during 43 days of hospitalization . Although the foal died at 3 months of age, serum creatinine concentration and electrolyte values remained within normal limits during the 3-month period. Spec Care Dentist, 1988 May-Jun, 8(3), 111 - 4 The relationship of oral Candida tropicalis infection to systemic candidiasis in a patient with leukemia; Redding SP et al.; Oropharyngeal candidiasis is an extremely common complication in patients receiving chemotherapy for leukemia . Candida tropicalis appears to be the major infectious agent when these patients develop candidemia . In this article, a case of C tropicalis fungemia with oropharyngeal manifestations is presented . The relationship of oropharyngeal candidiasis to oral candidal infection is discussed. EMBO J, 1988 Apr, 7(4), 1167 - 73 Acyl-CoA oxidase contains two targeting sequences each of which can mediate protein import into peroxisomes; Small GM et al.; Acyl-CoA oxidase is a major induced enzyme in peroxisomes of Candida tropicalis grown on fatty acids . The gene, POX4, encoding acyl-CoA oxidase was expressed in vitro, and the resulting polypeptide was imported into purified peroxisomes in a temperature-dependent fashion . Plasmids containing fragments of POX4 were prepared, expressed and the polypeptides tested for import into peroxisomes . We identified two regions of acyl-CoA oxidase (amino acids 1-118 and 309-427) that contained information that specifically targeted fragments of acyl-CoA oxidase to peroxisomes . The corresponding regions of the gene were fused to cDNA encoding the cytosolic enzyme dihydrofolate reductase (DHFR), and the expressed fusion proteins were likewise imported into peroxisomes . DHFR itself neither bound to, nor was imported into peroxisomes . Thus, there are at least two regions of peroxisomal targeting information in the acyl-CoA oxidase gene. J Med Vet Mycol, 1988 Apr, 26(2), 119 - 26 Polyamine depletion and growth inhibition in Candida albicans and Candida tropicalis by alpha-difluoromethylornithine and cyclohexylamine; Pfaller MA et al.; The ability of two known inhibitors of polyamine synthesis, alpha-difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase (ODC), and cyclohexylamine, an inhibitor of spermidine synthase, to inhibit the in vitro growth and polyamine synthesis of clinical isolates of Candida tropicalis and Candida albicans was examined . Treatment of C . tropicalis and C . albicans with either DFMO or cyclohexylamine resulted in depletion of cellular polyamines and inhibition of growth . The growth inhibition produced by each of these compounds was completely reversed by exogenous polyamines . Depletion of polyamines by low concentrations of DFMO significantly enhanced the growth inhibitory activity of cyclohexylamine versus C . albicans . DFMO inhibited ODC activity in both C . albicans and C . tropicalis . These findings document the ability of cyclohexylamine and DFMO to inhibit polyamine synthesis and growth in clinically important species of Candida. Biochemistry, 1988 Mar 8, 27(5), 1464 - 9 Stereochemical studies of D-glucal hydration by alpha-glucosidases and exo-alpha-glucanases: indications of plastic and conserved phases in catalysis by glycosylases; Chiba S et al.; Alpha-Glucosidases from Aspergillus niger, pig serum, ungerminated rice, buckwheat, and sugar beet seeds (but not from brewers' yeast or honeybee) were found to catalyze the hydration of D-glucal . Each reactive alpha-glucosidase, incubated with D-glucal in D2O, was shown to protonate (deuteriate) this prochiral substrate from above its re face, i.e., from a direction opposite that assumed for protonating alpha-D-glucosidic substrates . At the same time, D-glucal hydration catalyzed by three of the alpha-glucosidases that acted rapidly enough in D2O to determine product configuration was found to yield 2-deoxy-D-glucose of the same specific (alpha-) configuration as the D-glucose produced from alpha-D-glucosidic substrates . These findings substantially extend those reported earlier for the hydration of D-glucal by one (Candida tropicalis) alpha-glucosidase preparation . Together with other recent results, they suggest that the process of catalysis by alpha-glucosidases (and perhaps glycosylases in general) may comprise two separate and separately controlled parts, namely, a "plastic" phase concerned with substrate protonation and a substrate-unrelated "conserved" phase concerned with the creation of product configuration . In contrast to the alpha-glucosidases, three "inverting" exo-alpha-glucanases (Arthrobacter globiformis glucodextranase; Rhizopus niveus and Paecilomyces varioti glucoamylase) were found to protonate D-glucal from below its si face . Further, whereas the catalysis of D-glucal hydration by the alpha-glucosidases was intensively inhibited by excess substrate, that promoted by the exo-glucanases showed no detectable substrate inhibition. Prikl Biokhim Mikrobiol, 1988 Mar-Apr, 24(2), 218 - 25 {Study of the structure of mannans from Candida maltosa and Candida tropicalis using 13C-NMR spectroscopy}; Bovina EV et al.; Mannans were isolated from six Candida strains and characterized . 13C-NMR spectroscopy revealed interspecific and interstrain difference of the yeasts in the structure of their mannans. Infect Immun, 1988 Mar, 56(3), 601 - 6 Antigenic differences in the surface mannoproteins of Candida albicans as revealed by monoclonal antibodies; Sundstrom PM et al.; Monoclonal antibodies to Candida albicans were prepared with blastoconidia bearing germ tubes used as the immunogen . Four antibodies reacted by immunofluorescence with surfaces of C . albicans as well as Candida stellatoidea, Candida tropicalis, and several strains of C . albicans, but not with Torulopsis glabrata . One antibody reacted with Saccharomyces cerevisiae . In addition, the monoclonal antibodies precipitated material of approximately 200 kilodaltons when tested against metabolically labeled blastoconidia digests . The monoclonal antibodies exhibited heterogeneous staining of C . albicans surfaces, as shown by immunofluorescence . None of the monoclonal antibodies were specific to germ tubes . More importantly, however, two of the monoclonal antibodies reacted with the mannoprotein precipitin arc of C . albicans that was produced by reference rabbit polyclonal antisera by crossed immunoelectrophoresis, thus linking the heterogeneity seen by immunofluorescence to the heterogeneity in mannoproteins . Finally, three of the monoclonal antibodies reacted with a glycan fraction of cell digests, indicating their reactivity with the carbohydrate portion of the mannoprotein. FEBS Lett, 1988 Feb 29, 229(1), 215 - 8 Occurrence and possible roles of acetoacetyl-CoA thiolase and 3-ketoacyl-CoA thiolase in peroxisomes of an n-alkane-grown yeast, Candida tropicalis; Kurihara T et al.; Two kinds of 3-ketoacyl-CoA thiolases were found in the peroxisomes of Candida tropicalis cells grown on n-alkanes (C10-C13) . One was a typical acetoacetyl-CoA thiolase specific only to acetoacetyl-CoA, while another was 3-ketoacyl-CoA thiolase showing high activities on the longer chain substrates . A high level of the latter thiolase activity in alkane-grown cells was similar to that of other enzymes constituting the fatty acid beta-oxidation system in yeast peroxisomes . These facts suggest that the complete degradation of fatty acids to acetyl-CoA is carried out in yeast peroxisomes by the cooperative contribution of acetoacetyl-CoA thiolase and 3-ketoacyl-CoA thiolase. Chemioterapia, 1988 Feb, 7(1), 38 - 41 Comparative activity in different media of ketoconazole, miconazole and amphotericin B against Candida lusitaniae and sucrose-negative Candida tropicalis; Mignini F et al.; This study evaluates the susceptibility of sucrose-negative Candida tropicalis and Candida lusitaniae strains to amphotericin B (AMB), miconazole (MCZ) and ketoconazole (KTZ) . The susceptibility tests were carried out in different media: Antibiotic Medium 3 (AM-3m) and Earle Minimum Essential Medium (E-MEM) for AMB: Yeast Nitrogen Base (YNB) and E-MEM for imidazole compounds . The minimal fungicidal concentrations (MFCs) of AMB were slightly higher than minimal inhibitory concentration (MICs) except against Candida lusitaniae strains; whereas the MFCs of MCZ and KTZ were higher than the MICs by almost two-fold for all strains tested . AMB was more efficacious against sucrose-negative Candida tropicalis and the MICs were very definite; on the contrary, the MICs with KTZ were difficult to read . The MICs of AMB in E-MEM were essentially the same as those in AM-3m; whereas for KTZ and MCZ determined in YNB the MICs were generally higher than those obtained in E-MEM. Eur J Clin Microbiol Infect Dis, 1988 Feb, 7(1), 80 - 1 Comparative in vitro activity of LY121019 and amphotericin B against clinical isolates of Candida species; Spitzer ED et al.; LY121019 and amphotericin B were equally active in vitro against most clinical isolates of Candida albicans and Candida tropicalis . Higher concentrations of LY121019 were required for inhibition of Candida glabrata . Other Candida species were inhibited by amphotericin B but not by LY121019. Eur J Clin Microbiol Infect Dis, 1988 Feb, 7(1), 77 - 80 Evaluation of in vitro antifungal activity of LY121019; Hobbs M et al.; LY121019 is a new semisynthetic lipopeptide antifungal agent with potent in vitro fungicidal activity against multiple clinical strains of Candida albicans and Candida tropicalis but is 10-100 fold less active against Torulopsis glabrata and Candida parapsilosis . Its in vitro activity against Candida albicans and Candida tropicalis is comparable to that of amphotericin B . The in vitro fungicidal activity of this new agent supports further investigations into its use in treatment of Candida infections. Nahrung, 1988, 32(4), 327 - 33 Influence of selected sugars and temperature on fatty acids composition in Candida tropicalis; Tahoun MK et al.; Temperature, mono- and disaccharides affected lipids, biomass production as well as odd-chain and unsaturated fatty acids contents of Candida tropicalis . With various sugars as carbon sources at 30 degrees C, the order for biomass production was, galactose greater than glucose greater than sucrose greater than fructose greater than lactose, while lipids production/g biomass decreased as follows: galactose, glucose, sucrose, fructose and lactose . On the other hand, the odd-chain fatty acids contents decreased in the following order: fructose, sucrose, glucose and lactose . Lowering the temperature of cultivation to 15 degrees C, decreased biomass and lipids production . However, a notable decrease in odd-chain fatty acids contents was detected. Chemotherapy, 1988, 34(2), 96 - 100 In vitro synergistic activity of ketoconazole with trifluoperazine and with chlorpromazine against medically important yeasts; Ben-Gigi G et al.; Combination of ketoconazole and trifluoperazine or chlorpromazine yielded an in vitro synergistic effect on growth inhibition of Candida albicans, Torulopsis glabrata, Cryptococcus neoformans, Candida parapsilosis and Candida tropicalis . The optimal pH range for the synergistic effects was 7.0-7.6 . At pH 5.0 the drug combination was antagonistic . (Application for patent protection has been filed). Mol Gen Mikrobiol Virusol, 1988 Jan, (1), 29 - 33 {Identification of mutations in the asporogenic yeast Candida tropicalis using intrageneric fusion of protoplasts}; Golovina GI et al.; The possibility of genetic identification of mutations in asporogenic yeast by the technique of intrageneric fusion of yeast protoplasts of Candida tropicals and Saccharomyces cerevisiae has been demonstrated for Candida tropicals strains G5-9 (Ade- Leu-) and G32-4 (Leu-) . The mutations to auxotrophy ade- in the strain G5-9 and leu- in G32-4 of Candida tropicals are allelic to ade2 and leu1 mutations in the genes of Saccharomyces cerevisiae yeast . The allelic character of adenine auxotrophy mutation in Candida tropicals and ade2 mutation in Saccharomyces cerevisiae is confirmed by the absence of AIR-carboxylase activity in cellular extract from the strain G5-9. Acta Microbiol Hung, 1988, 35(3), 269 - 76 Changes of alternative respiration during the division cycle of Candida tropicalis; Novak B et al.; The changes in cyanide-resistant and SHAM-sensitive respiration have been examined in the cell cycle of yeast cells synchronized by nutrient starvation . The changing of the cyanide-resistant respiration of cells proliferating in synchronizing fermentor, where the glucose concentration was continuously decreasing, was associated with the stage in cell cycle rather than with the alteration of environment . The cyanide-resistant alternative respiration may be attributed characteristic of a certain part of the cell cycle termed "A" phase . This hypothesis is supported by the observation that in cultures proliferating, synchronously under constant conditions, the cyanide-resistant respiration changes periodically and reaches its maximum in the "A" phase after the cell division. Acta Microbiol Hung, 1988, 35(3), 259 - 68 The kinetics of cell division in a synchronizing fermentor; Novak B et al.; The kinetics of cell proliferation of Candida tropicalis synchronized by periodical glucose starvation was examined in a continuous synchronizing fermentor . The length of the deterministic (B) and stochastic (A) phase of the cell cycle was determined by cell density data analysis using the Transition Probability Model . The size distribution of the population shows that the absence of nutrient stops the cells at the beginning of the cell cycle but the succeeding cell divisions are not perfectly synchronized . The reason of this comes from the difference between the generation times of the cells which is the consequence of the different long "A" phases in their cycle. Microbiol Immunol, 1988, 32(10), 1013 - 24 Detection of specificity of a new antigen in Candida tropicalis and its evaluation by taxonomic DNA analyses; Hamajima K et al.; Monospecific factor serum for identifying Candida tropicalis was obtained either from rabbit antiserum to heated cells of C . tropicalis M 1519 (S 96) or from antiserum to C . tropicalis IFO 1400, by adsorption with heated cells of Candida albicans serotype A, or C . albicans (A) and Candida krusei, respectively . We designated this adsorbed serum factor t serum . The monospecific factor serum reacted with 31 out of 32 strains of C . tropicalis, only when tested on heat-treated cell antigens, whereas it did not react with any of 72 strains of the six other medically important species of Candida . The morphological and physiological characteristics of the one strain of C . tropicalis that did not react with the factor t serum, designated the t- -strain, were shown to be similar to those of the type strain of C . tropicalis by most of the methods employed for identifying Candida . Therefore, cell wall mannan from the t- -strain was compared with that from several typical strains of C . tropicalis for its specificity by the precipitation reaction and also for its 1H-nuclear magnetic resonance spectrum . The results showed that these mannans are similar to each other serologically and physicochemically, suggesting that the new antigen t is not mannan . Taxonomic characterization of the t-- and several typical strains of C . tropicalis was carried out by determining the mol% G+C of their DNA and also their DNA homology . Although the mol% G+C values of four typical strains of C . tropicalis were fairly similar (35.2 to 36.2 mol% by the Tm method and 35.5 to 36.4 mol% by the HPLC method), the t- -strain had a G+C content of 44.1 (Tm) and 43.3 (HPLC) mol% . Furthermore, the DNAs of the t- -strain and the type strain of C . tropicalis showed only 18.2% relatedness . These results suggest that the antigen corresponding to serum factor t exists only in the cell wall of C . tropicalis strains, not in those of the other medically important Candida, and that the t- -strain should not be classified as C . tropicalis . In conclusion, the taxonomic value and usefulness of factor t serum is primarily for differentiating C . tropicalis from C . albicans serotype A serologically. Eur J Biochem, 1987 Dec 30, 170(1-2), 105 - 10 Catalase gene of the yeast Candida tropicalis . Sequence analysis and comparison with peroxisomal and cytosolic catalases from other sources; Okada H et al.; A clone harbouring the genomic DNA sequence for the peroxisomal catalase of an n-alkane-utilizable yeast, Candida tropicalis, has been isolated by the hybrid-selection method and confirmed with a probe of catalase partial cDNA . Nucleotide sequence analysis of the cloned DNA disclosed that the gene fragment coding for catalase had a length of 1455 base pairs (corresponding to 485 amino acids; m = 54937 Da), and that the size of this enzyme was the smallest among all catalases reported hitherto . No intervening sequence was found in this coding region and some portions coincided with the amino acid sequences obtained from the analysis of the purified catalase . The comparison with three peroxisomal catalases from rat liver, bovine liver and human kidney, and one cytosolic catalase from Saccharomyces cerevisiae has revealed that catalase from C . tropicalis was more homologous to the peroxisomal enzymes than to the cytosolic one . C . tropicalis used the codons of the high-expression type . Amino acid residues were all conserved at the active and heme-binding sites . In the N and C-terminal regions there was no characteristic signal sequence or consensus sequence . However, a noticeable region, which can be discriminated between peroxisomal and cytosolic catalases, was proposed. Arch Intern Med, 1987 Dec, 147(12), 2117 - 20 Nosocomial fungemia in a large community teaching hospital; Harvey RL et al.; This report reviews 48 episodes of hospital-acquired fungemia that occurred over a four-year period at a large community teaching hospital . The incidence of hospital-acquired fungemia increased eightfold during the study period . Candida albicans (58%), Candida tropicalis (25%), and Candida parapsilosis (15%) were the most common fungal pathogens isolated from blood cultures . Twenty-one patients (44%) had concomitant bacteremia . Intravascular catheters (100%), antibiotic administration (98%), urinary catheters (81%), surgical procedures (65%), parenteral alimentation (60%), and corticosteroid administration (54%) were the most common predisposing factors . The overall mortality rate was 75% . Hospitalization on the medical service, age greater than 60 years, and hospital stay less than 100 days were associated with a significantly increased mortality rate. Eur J Clin Microbiol, 1987 Dec, 6(6), 628 - 33 Predictive value of surveillance cultures for systemic infection due to Candida species; Pfaller M et al.; Weekly fungal surveillance cultures (1,542 cultures) of urine (475), stool (520) and oropharyngeal (547) specimens from 111 patients on the bone marrow transplant and hematologic malignancy services were analyzed . Forty-three percent of the patients were colonized by Candida albicans and 10.8% by Candida tropicalis . There were 22 proven systemic fungal infections, ten due to Candida albicans, eight to Candida tropicalis, one each to Candida pseudotropicalis and Torulopsis glabrata, and two to Aspergillus species . Positive surveillance cultures for Candida tropicalis were highly predictive of systemic infection . The finding of two or more positive cultures yielded high positive predictive values (100%) as a function of body site . Positive surveillance cultures for Candida albicans were not predictive of disease but negative cultures for Candida albicans and Candida tropicalis had a high negative predictive value (95-99%) . Surveillance culture data for specific Candida species may aid in diagnostic and therapeutic decision making. Am J Clin Pathol, 1987 Dec, 88(6), 743 - 5 The use of the Prompt Inoculation System in preparing a standardized yeast inoculum; Pfaller MA et al.; One of the most important steps in performing broth dilution susceptibility testing of yeast isolates is the preparation of the starting inoculum . Although not specifically developed for yeast inoculum preparation, the Prompt Inoculation System (3M) provides a novel alternative approach that may provide a more standardized yeast inoculum than previously employed methods . The authors examined the relationship between the number of colonies picked with the Prompt Inoculation Wand and the hemacytometer and viable colony counts for each of six test organisms, including Candida albicans, Candida tropicalis, Candida krusei, Candida parapsilosis, Candida glabrata, and Cryptococcus neoformans . They found that by picking five colonies 1-2 mm in diameter, inoculum densities of 1 to 5 X 10(6) CFU/mL were obtained with most of the test organisms . A considerably higher inoculum density was observed with C . glabrata (1 to 2 X 10(7) CFU/mL) because of the small size of this organism . No overfilling of the Inoculation Wand was observed when more colonies were touched . This study indicates that the Prompt Inoculation System offers a convenient and simple method for yeast inoculum preparation. Eur J Biochem, 1987 Oct 1, 168(1), 245 - 50 Characterization of a soluble carnitine acetyltransferase from Candida tropicalis; Kozulic B et al.; Carnitine acetyltransferase was purified from the cytoplasmic fraction of Candida tropicalis grown on alkanes in continuous culture . By ion-exchange chromatography the enzyme was resolved in two fractions with the same specific activity of 80 U/mg . The molecular mass of both enzyme forms, determined by non-denaturing gradient gel electrophoresis, was 540 kDa . After SDS electrophoresis only one band of 64 kDa was detected indicating that both enzymes are oligomers each containing eight subunits . Isoelectric focusing in agarose under non-denaturing conditions demonstrated the presence of at least four different charged species in the pH range between 5.6 and 6.7 . After isoelectric focusing in 9 M urea/1% Nonidet P-40 gels, both enzyme forms were resolved into four bands . Peptide mapping, performed by cyanogen bromide cleavage of polypeptides separated by denaturing isoelectric focusing followed by second-dimension SDS electrophoresis, revealed a very high degree of homology between these polypeptides . The presence of the octameric form of carnitine acetyltransferase already in the starting material was demonstrated by non-denaturing gradient gel electrophoresis and immunoblotting . Antibodies against carnitine acetyltransferase from C . tropicalis ATCC 32113 formed precipitation lines with extracts from several Candida species but not with extracts of Candida utilis, Candida ethanothermophilum and an another strain of C . tropicalis. Carcinogenesis, 1987 Oct, 8(10), 1543 - 8 Possible mycological etiology of oral mucosal cancer: catalytic potential of infecting Candida albicans and other yeasts in production of N-nitrosobenzylmethylamine; Krogh P et al.; Yeasts were isolated from 12 cases of oral precancerous lesions (leukoplakia and erythroleukoplakia) by sampling the lesion as well as normal mucosa of each patient, yielding 21 strains of Candida albicans belonging to 15 biotypes, two strains of C . tropicalis, one strain of C . parapsilosis and two strains of Torulopsis glabrata . Biopsies were obtained from the lesions for histologic examination . The catalytic potential of the yeast strains to form N-nitrosobenzylmethylamine (NBMA) from the precursors N-benzylmethylamine and nitrite was assessed at pH 6.8 . The NBMA produced was identified and quantitated by h.p.l.c . and confirmed by g.c.-m.s . Nitrosation rates were calculated as total nitrosamine subtracted the chemically produced nitrosamine, and related to number of yeast cells . The yeast strains differed in nitrosation potential (P less than 0.001), ranking from 0 to 1.2 micrograms NBMA/10(6) cells . Candida albicans strains, belonging to the biotypes 051, 147, 151, 153, 157 and 353, which constitute more rarely occurring biotypes, exhibited the highest nitrosation potential . Candida tropicalis, C . parapsilosis and T . glabrata were ranked lower . Strains with high nitrosation potential were generally isolated from lesions with more advanced precancerous changes . The yeast cells were present in the superficial part of the epithelium of the lesions as branching mycelium, and in some cases extending from the mucosal surface to the deeper epithelial cell layers . This might represent a fungal transportation system which could channel precursors in the saliva at the mucosal surface to the deeper part of the epithelium where the produced nitrosamine could be deposited . Thus, further evidence is provided supporting the hypothesis that certain strains of C . albicans and of other yeasts play a causal role in the development of oral cancer, by means of endogenous nitrosamine production. Farmaco {Sci}, 1987 Oct, 42(10), 747 - 53 Antimycotic action of methyl substituted N-(5-pyrimidinyl)benzenesulfonamide derivatives; Pecorari P et al.; Some series of N-(5-pyrimidinyl)benzenesulfonamide variously methylated at the ring and/or sulfonamidic nitrogens and substituted at the benzene with NO2 or NH2 were synthesized and studied spectrometrically (N.M.R) . When tested on several strains of Candida albicans and Candida tropicalis, some of the compounds exhibited very slight antimycotic activity. Rev Infect Dis, 1987 Sep-Oct, 9(5), 1006 - 12 Mycoses caused by Candida lusitaniae; Hadfield TL et al.; Candida lusitaniae, a fungus with a low incidence of infection in immunocompetent people, is emerging as an opportunistic pathogen in immunocompromised hosts . This yeast is generally resistant to amphotericin B and may present therapeutic difficulties . C . lusitaniae may be misidentified as one of several other fungal species, including Candida tropicalis, Candida parapsilosis, and even Saccharomyces cerevisiae . As judged by in vitro antifungal susceptibility testing, minimal inhibitory concentrations of antifungal agents other than amphotericin B are achievable, but fungicidal levels are not . When encountered in blood or other body sites, C . lusitaniae should be carefully considered as a potential pathogen. Mycopathologia, 1987 Sep, 99(3), 155 - 71 Uncommon yeastlike zoopathogens and commercial systems for their identification; Land GA et al.; Opportunistic infections caused by yeasts or yeastlike fungi have increased in incidence in recent years as a result of clinical and therapeutic factors . Several formerly uncommon yeastlike zoopathogens--Candida lusitaniae, Candida paratropicalis (sucrose-negative variant of Candida tropicalis), Trichosporon beigelii, Blastoschizomyces capitatus, and Rhodotorula species--have been isolated from patients with invasive infections . The increased isolation of such opportunistic pathogens from a variety of clinical specimens has created a demand for simple, rapid, reliable, and accurate commercial systems to assist laboratorians in identification . Here we summarize the manual and automated systems currently available and present detailed descriptions of three representative commercial products, i.e., API 20C, Abbott Quantum II, and API Yeast-Ident. J Clin Microbiol, 1987 Sep, 25(9), 1701 - 4 Reaction of fungal products with amebocyte lysates of the Japanese horseshoe crab, Tachypleus tridentatus; Hodes DS et al.; A commercially available endotoxin assay (CS-TAL) employing a chromogenic peptide and an amebocyte lysate from the Japanese horseshoe crab, Tachypleus tridentatus, gave a positive result with aqueous extracts of all 15 strains of Candida albicans and 1 strain each of Candida tropicalis, Cryptococcus neoformans, and a Mucor species that we tested . Purified glucans prepared from the Candida strains gave the same results . Reconstruction experiments showed that the positive results were not due to contaminating endotoxin . By contrast, assays employing amebocyte lysates of the American horseshoe crab, Limulus polyphemus, were inconsistent . Japanese workers have presented evidence that glucans activate the Tachypleus amebocyte lysate system by acting on an enzyme different from that on which endotoxin acts . Using a Tachypleus lysate preparation (Endospecy; Seikagaku Kogyo, Tokyo, Japan) from which this enzyme was excluded, we demonstrated a 5- to 10-fold drop in reactivity to the aqueous Candida extracts and glucans, whereas reactivity to endotoxin was unchanged . Normal human plasma was shown to decrease the effect of fungal extracts on CS-TAL . This inhibition was completely removed by heating the plasma . Our results suggest that Tachypleus systems may be of use clinically in distinguishing bacterial from fungal infections. Biochem Biophys Res Commun, 1987 Aug 14, 146(3), 1311 - 7 Isolation of the Candida tropicalis gene for P450 lanosterol demethylase and its expression in Saccharomyces cerevisiae; Chen C et al.; We have isolated the gene for cytochrome P450 lanosterol 14 alpha-demethylase (14DM) from the yeast Candida tropicalis . This was accomplished by screening genomic libraries of strain ATCC750 in E . coli using a DNA fragment containing the yeast Saccharomyces cerevisiae 14DM gene . Identity of this gene was confirmed by a) observing a heme binding region common to all P450s after sequencing the 3' portion of the gene, and b) based upon tests of its expression in strains of Saccharomyces cerevisiae. FEBS Lett, 1987 Aug 10, 220(1), 31 - 5 Isolation of several cDNAs encoding yeast peroxisomal enzymes; Ueda M et al.; Several candidate clones carrying partial cDNAs for yeast peroxisomal enzymes, such as catalase, carnitine acetyltransferase, isocitrate lyase, malate synthase and acyl-CoA oxidase, were efficiently isolated at a single plating from a phage lambda gt11 recombinant cDNA library prepared with poly(A)-rich RNA from an n-alkane-grown yeast, Candida tropicalis, with a mixture of antibodies against the respective purified enzymes . Among them, one candidate clone carrying partial cDNA for catalase was subcloned and subjected to nucleotide sequence analysis . We succeeded in determining that the amino acid sequence deduced from the nucleotide analysis included the sequences derived from the two peptide fragments obtained from the purified enzyme. J Clin Microbiol, 1987 Aug, 25(8), 1416 - 20 Candida tropicalis infection in normal, diabetic, and neutropenic mice; Fromtling RA et al.; Opportunistic infections caused by Candida tropicalis have been noted with increasing frequency in compromised patients . The pathogenicity of three isolates of C . tropicalis was studied in normal CD-1 mice, streptozotocin-induced diabetic mice, and cyclophosphamide-induced neutropenic mice . Lethal dose 50% endpoints and quantitative distribution of yeast cells in spleen, liver, and kidneys of mice infected intravenously were determined . The virulence of one yeast isolate was greater than that of the other two . The order of susceptibility to mortality and degree of organ colonization was neutropenic greater than diabetic greater than normal . Renal lesions resembling those associated with infection by C . albicans appeared by day 5 postinfection in diabetic and neutropenic mice . Greater numbers of C . tropicalis yeast cells were isolated from homogenates of the affected kidneys, suggesting that the kidney is a target organ for this fungus . This study demonstrates the increased susceptibility of compromised mice to C . tropicalis as compared with normal mice and verifies the ability of these yeasts to cause opportunistic disease. J Bacteriol, 1987 Aug, 169(8), 3696 - 700 Kinetics and inhibition studies of catechol O-methyltransferase from the yeast Candida tropicalis; Veser J; The Kms for esculetin and S-adenosyl-L-methionine for catechol O-methyltransferase from the yeast Candida tropicalis were 6.2 and 40 microM, respectively . S-Adenosyl-L-homocysteine was a very potent competitive inhibitor with respect to S-adenosyl-L-methionine, with a Ki of 6.9 microM . Of the catechol-related inhibitors, purpurogallin, with a Ki of 0.07 microM, showed the greatest inhibitory effect . Sulfhydryl group-blocking reagents, such as thiol-oxidizing 2-iodosobenzoic acid and mercaptide-forming p-chloromercuribenzoic acid, provided evidence for sulfhydryl groups in the active site of the enzyme . Yeast catechol O-methyltransferase is a metal-dependent enzyme and requires Mg2+ for full activity . Zn2+ and Mn2+ but not Ca2+ were able to substitute for Mg2+ . Mn2+ showed optimal enzyme activation at concentrations 50- to 100-fold lower than those of Mg2+. J Cell Biol, 1987 Jul, 105(1), 247 - 50 Import of the carboxy-terminal portion of acyl-CoA oxidase into peroxisomes of Candida tropicalis; Small GM et al.; We report the sequence of a cDNA clone that codes for the carboxy-terminal portion of the peroxisomal protein, acyl-CoA oxidase, from the yeast, Candida tropicalis . This is a newly identified acyl-CoA oxidase sequence, most likely a second allele of POX4 . The cDNA clone was expressed by in vitro transcription followed by translation . The major product, a 43-kD protein, associated with isolated peroxisomes in an in vitro import assay . More than half of the peroxisome-associated protein was protected from added protease, implying that it was internalized within the organelle . These findings indicate that there is sufficient information in the carboxy-terminal portion of the protein to target it to peroxisomes. Biochim Biophys Acta, 1987 Jun 6, 909(1), 35 - 43 Isolation of cDNA clones coding for peroxisomal proteins of Candida tropicalis: identification and sequence of a clone for catalase; Rachubinski RA et al.; A cDNA library, complementary to mRNAs of alkane-grown Candida tropicalis, was screened by differential DNA dot-blot hybridization with {32P}cDNA reverse-transcribed from mRNA of alkane-grown cells or from cells in which peroxisome formation was repressed by growth on glucose . 9% of the library encodes alkane-induced sequences . The cell-free translation products of eight hybrid-selected mRNAs were characterized by SDS-polyacrylamide gel electrophoresis and fluorography: most of them are probably peroxisomal proteins . Among these, a catalase clone was identified by immunoprecipitation of the translation product with anti-catalase . The clone was sequenced: the inferred amino acid sequence is homologous to the carboxytermini of mammalian and Saccharomyces cerevisiae catalases . C . tropicalis catalase mRNA is 1.7-1.8 kb long by Northern analysis, of which 1.5-1.6 kb is required to code for the 57 kDa polypeptide . Catalase mRNA (assayed by dot-blot hybridization) is strikingly induced in C . tropicalis by growth on alkanes, suggesting that peroxisome induction is transcriptionally regulated . This sublibrary of alkane-induced, mostly peroxisomal clones, together with a recently developed cell-free peroxisome protein import assay, will permit investigation of the targeting of proteins to peroxisomes. Pharmazie, 1987 Jun, 42(6), 378 - 81 Synthesis and antimycotic activity of some benzyloxyimino compounds; Garuti L et al.; Some benzyloxyimio compounds, related to oxiconazole and having a 1H-indole or 1H-benzimidazole moiety, have been synthesized and tested in vitro for their antimycotic activity against Candida tropicalis and C . albicans . The most active was showed to be 0-(2,4-dichlorobenzyl)-1-benzyl-5-nitro-1H-benzimidazole-2-carboxaldehyd e oxime (MIC: 25 micrograms/ml against both microorganisms) . A structural feature important for the biological activity of the series appears to be presence of a benzimidazole nucleus substituted by an electron withdrawing group. Pathol Biol (Paris), 1987 Jun, 35(5 Pt 2), 879 - 81 {In vitro antifungal activity of nitroxoline . Preliminary clinical results}; Cancet B et al.; Nitroxoline is an oxyquinoline derivative with a large antifungical activity . The fungistatic activity of nitroxoline is greater against Candida albicans than against Torulopsis glabrata, Candida tropicalis and Candida krusei . The MIC are compatible with urinary concentrations of nitroxoline . These preliminary clinical results favor the use of nitroxoline in the management of fungal urinary tract infections. Infect Immun, 1987 Jun, 55(6), 1490 - 7 Flow cytometric analysis of the DNA synthetic cycle of Candida species; Dvorak JA et al.; The total DNA per cell and DNA synthetic cycle phases were determined by flow cytometry in five Candida isolates including three species: Candida albicans 208R1, Candida tropicalis ATCC 750, and Candida parapsilosis 970, 3138, and ATCC 22019 . The cells were prepared for flow cytometry by fixation in Carnoy fixative followed by staining with mithramycin . Marked but stable and reproducible inter- and intraspecific differences in total DNA per cell of stationary-phase cultures were found which did not correlate directly to diphenylamine estimates of the same parameter . This discrepancy was resolved by mathematically converting flow cytometry data into diphenylamine data . The reason for the discrepancy was found in studies of the DNA synthetic cycle of these yeasts: a large but isolate-specific variable proportion of the population is arrested in the S and G2-M phases after the culture passes from exponential to stationary phase . Histograms of exponential-growth-phase Candida isolates demonstrate that the majority of the population is in the G2-M phase of the DNA synthetic cycle . The DNA content of the C . tropicalis and C . parapsilosis isolates studied is as high as or higher than that of C . albicans . Extranuclear fluorescent particles were observed in the C . tropicalis isolate . No equivalent particles could be detected in the other four Candida isolates . The nature of the particles is unknown. Mol Cell Biol, 1987 May, 7(5), 1848 - 55 Efficient association of in vitro translation products with purified stable Candida tropicalis peroxisomes; Small GM et al.; Newly synthesized peroxisomal proteins enter preexisting peroxisomes posttranslationally in vivo, generally without proteolytic processing . An efficient reconstitution of this process in vitro together with cloned DNAs for peroxisomal proteins would make possible investigation of the molecular information that targets proteins to peroxisomes . We have previously reported the isolation of clones for Candida tropicalis peroxisomal proteins; here we describe the association (and possible import) of peroxisomal proteins with peroxisomes in vitro . C . tropicalis was grown in a medium containing Brij 35, resulting in the induction of a moderate number of medium-sized peroxisomes . These peroxisomes, isolated in a sucrose gradient, had a catalase latency of 54% and were sufficiently stable to be concentrated and used in an import assay . The reticulocyte lysate translation products of total RNA from oleate-grown cells were incubated with the peroxisomes at 26 degrees C in the presence of 50 mM KCl, protease inhibitors, 0.5 M sucrose, 2.5 mM MOPS (morpholinepropanesulfonic acid) (pH 7.2), and 0.5 mM EDTA . Ten major translation products (which could be immunoprecipitated with antiserum against peroxisomal protein) became progressively associated with the peroxisomes during the first 30 min of incubation (some up to approximately 70%) . These include acyl coenzyme A oxidase and the trifunctional protein hydratase-dehydrogenase-epimerase . This association did not occur at 4 degrees C nor did it occur if the peroxisomes were replaced with mitochondria. J Bacteriol, 1987 May, 169(5), 2284 - 6 Peroxisomal localization and activation by bivalent metal ions of ureidoglycolate lyase, the enzyme involved in urate degradation in Candida tropicalis; Takada Y et al.; Ureiodoglycolate lyase (EC 4.3.2.3) was found only in the peroxisomes in urate-induced Candida tropicalis . The enzyme was markedly activated by the bivalent metal ions Mn2+, Fe2+, and Ni2+ . The activation by Mn2+ was suggested to be the result of its binding to the apoenzyme. J Oral Pathol, 1987 May, 16(5), 282 - 4 Carriage of Candida species in the oral cavity in diabetic patients: relationship to glycaemic control; Fisher BM et al.; To study the possible relationship between the quality of glycaemic control in diabetes mellitus and the carriage of Candida species, the candidal carrier status of 412 diabetic patients was examined using an oral rinse technique and correlated with measurements of random blood glucose and total glycosylated haemoglobin . Candida was isolated in 210 diabetics (51%) with 13 patients (6%) carrying more than one species . The positive isolates were: Candida albicans (89%), Candida krusei (2.8%), Candida glabrata (2.8%), Candida tropicalis (6.2%), Candida stellatoidea (2.8%) and Candida parapsilosis (0.5%) . No association was identified between carriage rates and the type of treatment of diabetes, or with the quality of glycaemic control . As in non-diabetic subjects, the carriage rates were higher in diabetic patients wearing dentures . Thus, the oral carriage of Candida in diabetic patients was independent of glycaemic control but in certain sub-groups the carriage rates were higher, and involved uncommon candidal species. Biochem Biophys Res Commun, 1987 Apr 14, 144(1), 251 - 7 Isolation of the alkane inducible cytochrome P450 (P450alk) gene from the yeast Candida tropicalis; Sanglard D et al.; The gene for the alkane-inducible cytochrome P450, P450alk, has been isolated from the yeast Candida tropicalis by immunoscreening a lambda gt11 library . Isolation of the gene has been identified on the basis of its inducibility and partial DNA sequence . Transcripts of this gene were induced by alkane to levels 500 to 1000 fold over those detected in glucose-grown cells . The nucleotide sequence of the 3' portion of this gene revealed a coding sequence for the heme binding segment characteristic of the P450 gene family. Infect Immun, 1987 Mar, 55(3), 541 - 6 Candidacidal factors in murine bronchoalveolar lavage fluid; Nugent KM et al.; Respiratory secretions provide an efficient method for protecting the large surface area of the lower respiratory tract . To determine whether lung secretions contribute to antifungal defenses, we tested bronchoalveolar lavage fluid for fungicidal activity . Candida albicans (blastoconidia) was incubated in unconcentrated cell-free lavage fluid from Swiss Webster mice and then cultured quantitatively to measure residual viability . In control buffer the residual fractions of viable fungi were 1.03 +/- 0.12 at 60 min and 0.84 +/- 0.05 at 120 min, whereas the residual fractions in lavage fluid were 0.64 +/- 0.07 and 0.23 +/- 0.05, respectively (P less than 0.05 by t tests) . This activity was trypsin sensitive and heat stable (56 degrees C) and did not require divalent cations . It did not sediment with the surfactant fraction of lung lavage fluid . Unconcentrated lavage fluid reduced the adherence of C . albicans to serum-coated glass tubes to 2.3 +/- 1.5% of that of control Candida suspensions (n = 5, P less than 0.05 by t test) . It did not alter Candida ingestion or intracellular processing by alveolar macrophages . Lavage fluid also killed clinical isolates of Candida tropicalis and Torulopsis glabrata but did not kill Candida krusei or Candida parapsilosis . Lavage fluid was concentrated and passed through an acrylamide-agarose gel matrix . The chromatogram indicated that the candidacidal activity eluted in a peak with a molecular weight range of 29,000 to 40,000 . After electrophoresis on 15% sodium dodecyl sulfate-polyacrylamide gels, these fractions resolved into three bands . These were transferred to nitrocellulose and then eluted with Triton X-100; this procedure permitted the isolation of a single band of candidacidal activity with a molecular weight of 29,000 . In summary, murine lavage fluid contains a heat-stable protein with direct antifungal activity . This soluble factor may contribute to lung defense processes by reducing fungal viability and adherence to tissue surfaces. Antimicrob Agents Chemother, 1987 Mar, 31(3), 421 - 9 Effect of lipid composition and liposome size on toxicity and in vitro fungicidal activity of liposome-intercalated amphotericin B; Szoka FC Jr et al.; Intercalation of amphotericin B into liposomes at a 10 mol% drug/lipid ratio decreased its cytotoxicity by 3- to 90-fold in cultured murine cells and reduced its lethality by 2- to 8-fold in a median lethal dose (LD50) test in mice when compared with the commercial deoxycholate-solubilized drug (LD50 = 2.3 mg/kg) . The cytotoxicity and lethality of the liposomal preparations were a function of their lipid composition and diameter . There was no correlation between the reduction of toxicity in the tissue culture assay and the reduction of lethality in the LD50 test . The rank order of reduction of lethality was sterol-containing liposomes greater than solid liposomes greater than fluid liposomes . In general, small sterol-containing vesicles were less lethal than large vesicles of the same composition . Intercalation of amphotericin B in sterol or solid liposomes increased not only the LD50 but also the time to death . The organ distribution of amphotericin B 24 h after intravenous administration was similar whether the drug was given as the commercial deoxycholate preparation or in liposomes . Finally, there were no differences among any of the formulations in their fungicidal activity against Candida tropicalis and Saccharomyces cerevisiae in vitro . The lesser and slower lethality of the liposomal and detergent-solubilized drug suggests that the mechanism by which liposomes reduce the lethality of amphotericin B is by slowing its rate of transfer to a sensitive cellular target. Rev Infect Dis, 1987 Mar-Apr, 9(2), 398 - 402 Rapid diagnosis of candidiasis and aspergillosis; Bennett JE; Published studies support the hypothesis that at least two antigens of Candida albicans and Candida tropicalis circulate in the bloodstream of patients with severe candidiasis . One antigen (probably mannan) is stable and the other (probably protein) is labile . The stabile antigen can rarely be detected without prior dissociation from antibody . Dissociation treatment destroys the labile antigen, leaving in doubt whether any is antibody bound . Dissociation steps have also been necessary for detection of Aspergillus fumigatus antigen in sera from patients with invasive aspergillosis . Concentrations of the stable antigens of both Candida and Aspergillus appear to be in the nanogram-per-milliliter range, generally lying at the limits of detection by conventional assays . Improvements in sensitivity and practicality are needed, but the tests are clearly promising. J Clin Microbiol, 1987 Mar, 25(3), 563 - 6 Molecular probe for identification of medically important Candida species and Torulopsis glabrata; Mason MM et al.; A cloned DNA fragment from Candida albicans containing the gene for the protein actin was used to probe the molecular structure of the actin gene of several medically important yeasts (C . albicans, Candida stellatoidea, Candida tropicalis, Candida pseudotropicalis, Candida krusei, Candida parapsilosis, Candida guilliermondii, and Torulopsis glabrata) . Whole-cell DNA from each species was digested with restriction endonucleases, electrophoresed on agarose gels, and transferred to nitrocellulose . Radioactively labeled C . albicans actin gene was hybridized to the DNA fragments on the nitrocellulose . The C . albicans probe produced a strong signal with all of the Candida DNAs tested, indicating considerable conservation of this gene . In addition, the actin genes of all of the species tested were found to have no internal EcoRI or SalI restriction sites . With the exception of C . guilliermondii, all of the species tested had a single internal HindIII recognition site . However, the location of flanking restriction sites was found to be species specific . For all of the enzymes tested, the locations of the flanking restriction sites in C . albicans and C . stellatoidea were identical; all of the other strains yielded fragments clearly distinct from one another . These differences provide a molecular tool for the differentiation of medically important Candida species. Jpn J Antibiot, 1987 Feb, 40(2), 271 - 83 {Laboratory and clinical