Candida albicans

Transfusion, 1995 Jul, 35(7), 605 - 11
Protection of human polymorphonuclear leukocyte function from the deleterious effects of isolation, irradiation, and storage by interferon-gamma and granulocyte-colony-stimulating factor; Rex JH et al.; BACKGROUND: Fungal infections represent a difficult challenge to clinicians caring for neutropenic patients with hematologic malignancies, as antifungal therapy often has limited success in that setting . One promising yet problematic alternative approach is leukocyte transfusion . The isolation of polymorphonuclear leukocytes (PMNs) induces apoptosis and functional deterioration, and irradiation to prevent transfusion-associated graft-versus-host disease causes further functional deterioration . STUDY DESIGN AND METHODS: The ability of interferon-gamma and granulocyte-colony-stimulating factor (G-CSF), used both alone and in combination, to protect PMNs after 0 or 20 hours' storage in cell culture (as a model for function after transfusion) and irradiation with 0, 5, or 30 Gy was studied . RESULTS: Without cytokine treatment, 20-hour-old PMNs showed marked apoptosis, no appreciable chemotaxis, and no ability to kill Candida albicans . In contrast, cytokine treatment significantly reduced apoptosis and protected chemotaxis, C . albicans killing, and surface-receptor expression from both storage and irradiation . Although the majority of the benefit appeared to be due to G-CSF, consistent trends suggested better function of PMNs after combined treatment with interferon-gamma and G-CSF . CONCLUSION: Judicious use of cytokines may preserve PMN function . These findings have important implications for the transfusion of PMNs to cytopenic patients.

J Bacteriol, 1995 Jul, 177(13), 3788 - 92
Covalent association of beta-1,3-glucan with beta-1,6-glucosylated mannoproteins in cell walls of Candida albicans; Kapteyn JC et al.; Yeast and hyphal walls of Candida albicans were extracted with sodium dodecyl sulfate (SDS) . Some of the extracted proteins reacted with a specific beta-1,6-glucan antiserum but not with a beta-1,3-glucan antiserum . They lost their beta-1,6-glucan epitope after treatment with ice-cold aqueous hydrofluoric acid, suggesting that beta-1,6-glucan was linked to the protein through a phosphodiester bridge . When yeast and hyphal walls extracted with SDS were subsequently extracted with a pure beta-1,3-glucanase, several mannoproteins that were recognized by both the beta-1,6-glucan antiserum and the beta-1,3-glucan antiserum were released . Both epitopes were sensitive to aqueous hydrofluoric acid treatment, suggesting that beta-1,3-glucan and beta-1,6-glucan are linked to proteins by phosphodiester linkages . The possible role of beta-glucans in the retention of cell wall proteins is discussed.

J Infect Dis, 1995 Jul, 172(1), 192 - 8
Nitric oxide enhances resistance of SCID mice to mucosal candidiasis; Vazquez-Torres A et al.; The capacity of macrophages from SCID and C.B-17 mice to kill Candida albicans via a nitric oxide (NO)-dependent pathway and the contribution of NO in resistance to mucosal candidiasis were assessed . In vitro, an inhibitor of NO synthase (NOS) reduced the candidacidal activity and nitrite-producing capacity of activated resident peritoneal macrophages from immunocompetent C.B-17 and immunodeficient SCID mice . In vivo, stomachs from gnotobiotic SCID mice that were colonized with a pure culture of C . albicans had low-grade infections and expressed inducible NOS (iNOS) mRNA . C . albicans-monoassociated SCID mice treated with an inhibitor of NOS had more severe orogastric candidiasis than controls . These data suggest that NO contributes to the candidacidal capacity of activated macrophages from C.B-17 and SCID mice and that NO synthesized by iNOS may contribute to the resistance of SCID mice to mucosal candidiasis.

Turk J Pediatr, 1995 Jul-Sep, 37(3), 247 - 52
Amphotericin B in the treatment of candida meningitis in three neonates; Aydin M et al.; Candidiasis is an opportunistic infection and may result in significant morbidity and mortality in neonates . Cerebral candidiasis is rare and usually associated with systemic candidiasis . Information concerning the toxicity and efficacy of antifungal therapy for neonates is limited . In this report, we present three neonates with candidiasis . All of the patients were premature with low birth weights, and received antibiotic therapy for one to four weeks before the onset of candidiasis . Candida albicans was isolated from cerebrospinal fluid cultures . Amphotericin B was given administered at an initial dose of 0.25 mg/kg/day intravenously (IV) and increased to a dosage of 2 mg/kg/day, and therapy was continued for three to four weeks . A transient and mild elevation in hepatic enzyme concentration was observed in two patients, and transient thrombocytopenia occurred in all of them.

Antimicrob Agents Chemother, 1995 Jul, 39(7), 1538 - 41
Inhibition of sterol 4-demethylation in Candida albicans by 6-amino-2-n-pentylthiobenzothiazole, a novel mechanism of action for an antifungal agent; Kuchta T et al.; The effects of 6-amino-2-n-pentylthiobenzothiazole (APB), a new antifungal agent, on ergosterol biosynthesis in Candida albicans and Saccharomyces cerevisiae were studied, using {14C}acetate incorporation . In C . albicans, the inhibition of growth was accompanied by a marked inhibition of acetate incorporation in 4-desmethylsterols, with a significant portion of the radiolabel being incorporated in 4,4-dimethylsterols, lanosterol, and 4,4-dimethylzymosterol and minor amounts being incorporated in 4-methylsterols and squalene . The data are interpreted as evidence of a block of the ergosterol biosynthesis pathway at the level of 4-demethylation of 4,4-dimethylzymosterol, with partial inhibition of lanosterol 14-dimethylation and squalene epoxidation also being possible . In 6-amino-2-n-pentylthiobenzothiazole-treated S . cerevisiae, a significant amount of the radiolabel was incorporated also in 4-methylsterols, 4-methylzymosterol, and 4-methylfecosterol, indicating that in this microorganism there are different sensitivities of the two 4-demethylations and that the pathway is blocked at the level of 4-demethylation of 4-methylsterols.

Antimicrob Agents Chemother, 1995 Jul, 39(7), 1512 - 6
Patterns of in vitro activity of itraconazole and imidazole antifungal agents against Candida albicans with decreased susceptibility to fluconazole from Spain; Martinez-Suarez JV et al.; Two groups of recent clinical isolates of Candida albicans consisting of 101 isolates for which fluconazole MICs were < or = 0.5 microgram/ml (n = 50) and > or = 4.0 micrograms/ml (n = 51), respectively, were compared for their susceptibilities to fluconazole, clotrimazole, miconazole, ketoconazole, and itraconazole . Susceptibility tests were performed by a photometer-read broth microdilution method with an improved RPMI 1640 medium supplemented with 18 g of glucose per liter (RPMI-2% glucose; J . L . Rodriguez-Tudela and J . V . Martinez-Suarez, Antimicrob . Agents Chemother . 38:45-48, 1994) . Preparation of drugs, basal medium, and inocula was done by the recommendations of the National Committee for Clinical Laboratory Standards . The MIC endpoint was calculated objectively from the turbidimetric data read at 24 h as the lowest drug concentration at which growth was just equal to or less than 20% of that in the positive control well (MIC 80%) . In vitro susceptibility testing separated azole-susceptible strains from the strains with decreased susceptibilities to azoles if wide ranges of concentrations (20 doubling dilutions) were used for ketoconazole, miconazole, and clotrimazole . By comparison with isolates for which fluconazole MICs were < or = 0.5 microgram/ml, those isolates for which fluconazole MICs were > or = 4.0 micrograms/ml were in general less susceptible to other azole drugs, but different patterns of decreased susceptibility were found, including uniform increases in the MICs of all azole derivatives, higher MICs of several azoles but not others, and elevated MICs of fluconazole only . On the other hand, decreased susceptibility to any other azole drug was never found among strains for which MICs of fluconazole were lower.

Sex Transm Dis, 1995 Jul-Aug, 22(4), 221 - 7
Screening for Chlamydia trachomatis infection in pregnant women in Martinique; Chout RT et al.; GOAL OF THIS STUDY: To determine the prevalence of Chlamydia trachomatis urogenital infection and to identify behavioral, demographic, and clinical factors associated with the infection in pregnant women in Martinique . STUDY DESIGN: One-thousand-four-hundred-eleven patients 15-39 years old, at 10-16 weeks of gestation and attending the prenatal clinic at Lamentin Hospital, were tested for Chlamydia trachomatis infection of the cervix and urethra using tissue culture . RESULTS: Chlamydia trachomatis was isolated from 375 (26.7%) women; 34% of them were positive in the cervix and urethra, 58% in the cervix only, and 8% in the urethra only . Factors found by multivariate analysis to be significantly associated with chlamydial infection were age less than 25 years, first intercourse at less than 18 years old, previous induced abortions, mucopurulent cervicitis, and repeated candidiasis . CONCLUSIONS: None of the factors associated with chlamydial infection was sensitive enough to permit efficient selective screening . It is cost effective to recommend a routine screening for chlamydial infection together with an educational programPIP: To determine the prevalence of Chlamydia trachomatis urogenital infection among pregnant women in Martinique, 1411 consecutive women presenting to Lamentin Hospital for their initial prenatal visit between 1988-90 underwent specimen collection and extensive interviews . The mean age of study subjects was 27.1 years and the mean number of life-time sex partners was 3.2 . C . trachomatis was isolated from 375 women (26.7%), two-thirds of whom were asymptomatic . There was an inverse correlation between age and infection rate; 164 (43.7%) infected women were under 25 years of age . 34% had evidence of infection in both the cervix and urethra, 58% in the cervix only, and 8% in the urethra only . Other sexually transmitted pathogens with a high prevalence in this group included Ureaplasma urealyticum (39.9%), Candida albicans (32%), and Trichomonas vaginalis (13.7%) . Factors that correlated significantly with chlamydia infection by multivariate analysis were age less than 25 years, first intercourse less than 18 years, previous induced abortion, cervicitis, and repeated candidiasis . However, no single risk factor or constellation of risk factors was sufficiently sensitive to form the basis of a selective screening program . Considering the serious maternal and infant complications of C . trachomatis infection, routine screening in pregnant women is urged . Given a prevalence rate of 27%, 1630 infected pregnant women should be identified each year in Martinique . The cost of screening and treating these women and their partners would be US$250,000 compared to $1.2 million required to treat chlamydia-related conjunctivitis and pneumonia in infants and postpartum salpingitis in mothers .

Rev Fr Gynecol Obstet, 1995 Jul-Sep, 90(7-9), 345 - 51
{A comparative study of 2 ways of clinical management in premature rupture of the membranes at term: temporization versus labor induction}; Kouam L et al.; Two defined management approaches, temporization limited to 48 hours and immediate induction of labor, for premature rupture of the membranes at term were compared in a prospective study between January 1 1991 and November 30 1993 in the Maternity Unit of Yaounde University Hospital . During this period, 268 cases of premature rupture of the membranes were seen among 3252 deliveries, i.e . an incidence of 8.2% . In the temporization group (153 cases), spontaneous onset of labor was effective in 95 patients (62.1%) within 12 hours and in 137 patients (89.5%) within 24 hours after premature rupture of the membranes . Spontaneous deliveries in this temporization group accounted for 129 cases (92.8%) . In the induction of labor group, spontaneous delivery occurred in 119 cases (93.2%) . There were ten cesareans in the temporization group and eight cesareans and two vacuum cup extractions in the induction group . Short term (24 hours) prophylactic antibiotics were given to 34 patients, i.e . 16 cases in which the duration of rupture of the membranes was more than 24 hours and 18 cases of cesarean section . Maternal infections concerned 18 cases (6.7%) including 12 cases (4.4%) of malaria . Microbiology of vaginal swabs revealed 6 cases of pseudomonas, 4 cases of staphylococcus aureus and 3 of candida albicans . Neonatal infections confirmed by blood culture and assay of C-reactive-protein involved 24 cases (20.3%) . There were three fetal deaths, i.e . perinatal mortality of 1.1% . Risk factors, in these three fetal deaths, included postmaturity (1 case).(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemistry, 1995 Jun 20, 34(24), 7896 - 903
Mechanism of irreversible inactivation of phosphomannose isomerases by silver ions and flamazine; Wells TN et al.; Silver ions and silver-containing compounds have been used as topical antimicrobial agents in a variety of clinical situations . We have previously shown that the enzyme phosphomannose isomerase (PMI) is essential for the biosynthesis of Candida albicans cell walls . In this study, we find that PMI can be inhibited by silver ions . This process is shown to be irreversible, and is a two-step process, involving an intermediate complex with a dissociation constant, Ki, of 59 +/- 8 microM, and a maximum rate of inactivation of 0.25 +/- 0.04 min-1 in 50 mM Hepes buffer, pH 8.0 at 37 degrees C . The enzyme can be protected against this inactivation by the substrate mannose 6-phosphate, with a dissociation constant of 0.31 +/- 0.04 mM, close to its Km value . Flamazine (silver sulfadiazine) is a silver-containing antibiotic which is used clinically as a topical antimicrobial and antifungal agent . We compared the ability of silver sulfadiazine and two other silver-containing compounds to irreversibly inactivate C . albicans PMI . The addition of the organic moiety increased the affinity of the compounds, with silver sulfadiazine showing a Ki of 190 +/- 30 nM . In all cases, the maximum inhibition rate was similar, implying a similar rate-determining step . Silver sulfadiazine does not inhibit Escherichia coli PMI, and this suggests a role of the only free cysteine, Cys-150, in the inactivation process . To confirm this, we mutated this residue to alanine in C . albicans PMI . The resultant Cys150 --> Ala mutant protein showed similar Vm and Km values to the wild-type enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)

FEMS Microbiol Lett, 1995 Jun 15, 129(2-3), 103 - 13
Adherence of Candida albicans to host cells; Pendrak ML et al.; Research devoted to uncovering the mechanisms of adherence of Candida albicans to human tissue is reviewed . The physical aspects of adherence of the fungus to host cells and the biochemical and molecular features, as far as they are known, are discussed . Relevant pre- and post-adherence events in the pathogenesis of disease caused by this fungus are also noted . Putative adhesins and surface receptors of C . albicans for host proteins are discussed in detail.

J Leukoc Biol, 1995 Jun, 57(6), 842 - 50
Relationship of phospholipase C- and phospholipase D-mediated phospholipid remodeling pathways to respiratory burst activation in human neutrophils stimulated by Candida albicans hyphae; Meshulam T et al.; Neutrophil (PMN) oxidant release, a key component of defenses against disseminated candidiasis, was preceded by oxidant generation after stimulation by Candida albicans hyphae . Opsonized or unopsonized hyphae triggered phospholipase D (PLD) activation within 5 or 30 s, respectively, forming 1-O-alkyl-phosphatidic acid (alkyl-PA) or 1-O-alkyl-phosphatidyl-ethanol in the presence of ethanol . Ethanol, which competitively lowers phosphatidic acid (PA) production, caused dose-dependent inhibition of superoxide (O2-) generation after hyphal stimulation but altered neither baseline-unstimulated O2- production nor responses to phorbol myristate acetate . PA rises evoked by unopsonized hyphae began 2 min before significant O2- release, also preceding both phospholipase C activation and cytosolic Ca2+ rises . Diacylglycerol (DAG) rose in two distinct phases after stimulation by opsonized or unopsonized hyphae, peaking briefly after 60 or 120 s, respectively, followed by prolonged secondary rises . Initial DAG rises preceded inositol triphosphate elevations evoked by unopsonized hyphae . Though PA rose before DAG, no dephosphorylation of PA to form 1-O-alkyl-DAG was noted . Propranalol, which increases PA accumulation by inhibiting PA phosphohydrolase, lowered PMN O2- responses to hyphae . Early DAG rises temporally overlapped respiratory burst initiation but PMN responses to hyphae were unchanged by a DAG kinase inhibitor, R59022, which blocks phosphorylation of DAG to PA and enhances DAG accumulation . Thus, neither PA nor DAG accumulation individually accounted for triggering PMN O2- responses to hyphae . PLD activation and PA production may facilitate PMN fungicidal responses to hyphae but play an indirect role in initiating the respiratory burst.

J Clin Endocrinol Metab, 1995 Jun, 80(6), 1948 - 55
Priming of human polymorphonuclear neutrophilic leukocytes by insulin-like growth factor I: increased phagocytic capacity, complement receptor expression, degranulation, and oxidative burst; Bjerknes R et al.; Insulin-like growth factor I (IGF-I) is a GH-dependent peptide regulating mammalian growth that seems to be of importance for the normal development and function of the immune system . Polymorphonuclear neutrophilic leukocytes (PMNLs) are terminally differentiated phagocytes essential for host defense, and in the present study, recombinant human IGF-I was shown to be a powerful primer of mature human PMNLs . IGF-I augmented the PMNL phagocytosis of both immunoglobulin G-opsonized Staphylococcus aureus and complement-opsonized Candida albicans . In addition, the growth factor increased PMNL complement receptor expression {complement receptors 1 (CD35) and 3 (CD11b)} and primed the cells to stronger f-met-leu-phe-induced degranulation of both specific and azurophilic granules {markers: CD11b, CD35 and CD67 (specific granules); CD63 (azurophilic granules)} . In contrast, IGF-I did not alter the PMNL surface expression of Fc gamma RI (CD64), Fc gamma RII (CDw32), or Fc gamma RIII (CD16) . PMNLs exposed to IGF-I increased their f-met-leu-phe and phorbol myristate acetate-induced oxidative burst, as evaluated by hydrogen peroxide production, whereas IGF-I did not influence PMNL actin polymerization . The priming of PMNLs by IGF-I was dependent on time and concentration, and saturating amounts of a monoclonal antibody to the IGF-I receptor blocked the priming of PMNLs by this peptide . These experiments demonstrate that IGF-I can selectively stimulate mature PMNL functions, providing further evidence for the interaction between the immune and the endocrine systems.

Clin Exp Immunol, 1995 Jun, 100(3), 419 - 24
HIV-specific lymphoproliferative responses in asymptomatic HIV-infected individuals; Pontesilli O et al.; In vitro lymphoproliferative responses to HIV-1 recombinant antigens (gp160, p24, and Rev protein) were studied in 83 patients with asymptomatic HIV-1 infection (CDC groups II and III) and circulating CD4 lymphocyte numbers > 400/mm3 . Significant response to at least one of the three antigens was detected in 52.4% of the subjects, but the responses were weak, and concordance of the response to the three antigens was rare, the frequency of individuals responding to each antigen not exceeding 22.4% . Increasing frequencies of response were observed when recall antigens (tetanus toxoid and Candida albicans glycomannoprotein) (65.5%) and anti-CD3 MoAb (76.6%) were used as stimuli . Although a significant association between lymphocyte response to p24, but not gp160, and steadiness of CD4 lymphocyte numbers before the assay was observed, no predictive value for lack of CD4 cell decrease was confirmed for either antigen, and fluctuation of the responses to HIV antigens was seen during subsequent follow up . The panel of T cell assays used could be regarded as appropriate for monitoring both HIV-specific responses and T lymphocyte function during immunotherapy with soluble HIV antigens.

Obstet Gynecol, 1995 Jun, 85(6), 993 - 8
Torulopsis glabrata vaginitis; Spinillo A et al.; OBJECTIVE: To study the sociodemographic risk factors and clinical features of Torulopsis glabrata vaginal infection . METHODS: We evaluated the sociodemographic and clinical characteristics of 86 consecutive symptomatic women attending a vaginitis clinic and isolated T glabrata . Case patients were compared with a control group of 174 asymptomatic women with negative vaginal cultures and an additional group of 625 symptomatic women with vaginal cultures positive for Candida albicans . In addition, the sensitivity of the isolates to the more common antimycotic agents used was tested by the modified Kirby-Bauer method . RESULTS: Patients with T glabrata vaginal infection were from lower socioeconomic backgrounds and had less education . They were more likely to use vaginal tampons and to be seropositive for human immunodeficiency virus than were negative controls . Compared with C albicans infection, T glabrata was more frequent among women over 38 years of age and in those with less education and of lower social class . In logistic regression analysis, T glabrata was associated more frequently with recurrent vaginal candidiasis than was C albicans (odds ratio 2.46, 95% confidence interval 1.33-4.54; P = .004) . Six of the 86 (7%) T glabrata isolates and none of the C albicans isolates (P < .001 by Fisher exact test) were resistant to the imidazole derivatives tested . CONCLUSION: Torulopsis glabrata was isolated in 10% of women with vulvovaginal candidiasis attending a vaginitis clinic . This infection was associated with recurrent vaginitis in almost one-third of case patients presenting with symptoms.

J Infect Dis, 1995 Jun, 171(6), 1664 - 7
Defective killing of Candida albicans hyphae by neutrophils from beige mice; Jones-Carson J et al.; The candidacidal activity of neutrophils from BALB/c, C.B-17 +/+, C.B-17 scid/scid, scid/scid-bg/bg, C57BL/6 bg/+, C57BL/6 bg/bg, NIH III bg/bg-nu/+, and bg/bg-nu/nu mice was assessed . Killing of blastoconidia was moderately deficient with neutrophils from 2 strains of homozygous beige mice; however, neutrophils from all homozygous beige strains had a significantly decreased capacity to kill Candida albicans hyphae . This is the first demonstration of a decreased capacity of beige mouse neutrophils to kill C . albicans hyphae . The latter defect could be related to the enhanced susceptibility of beige mice to candidiasis.

J Infect Dis, 1995 Jun, 171(6), 1545 - 52
Efficacy and safety of fluconazole prophylaxis for fungal infections after marrow transplantation--a prospective, randomized, double-blind study; Slavin MA et al.; A randomized, double-blind, placebo-controlled trial assessed the efficacy and toxicity of 400 mg/day fluconazole in preventing fungal infections during the first 75 days after marrow transplantation . During prophylaxis, systemic fungal infections occurred in 10 (7%) of 152 fluconazole-treated patients compared with 26 (18%) of 148 placebo-treated patients (P = .004) . There were no Candida albicans infections in fluconazole recipients compared with 18 in placebo recipients (P < .001) and no significant increase in Candida infections other than C . albicans . Fluconazole also significantly reduced the incidence of superficial fungal infections (P < .001), fungal colonization (P = .037), and empiric amphotericin B use (P = .005) . The probability of survival was improved in fluconazole recipients, in whom 31 deaths occurred up to day 110 after transplantation compared with 52 deaths in placebo recipients (P = .004) . No clinically significant toxicity was detected with fluconazole use . Prophylactic fluconazole was safe and significantly reduced systemic fungal infections with other benefits, including improved survival at day 110 after marrow transplantation.

J Infect Dis, 1995 Jun, 171(6), 1539 - 44
Mortality of Candida albicans-infected mice is facilitated by superinfection of Escherichia coli or administration of its lipopolysaccharide; Akagawa G et al.; Pathogenesis of complex infection with Candida albicans and gram-negative bacteria was studied by determining the influence of infection with Escherichia coli or injection of E . coli lipopolysaccharide (LPS) on mortality of C . albicans-infected mice . Mice were infected intravenously with lethal doses of C . albicans, then treated intravenously at various times with viable E . coli or E . coli LPS, which individually were not lethal . Treatments 3 h after C . albicans infection clearly facilitated the death of the mice . Corresponding to this facilitated death, production of tumor necrosis factor (TNF) in sera was augmented 2 h after LPS injection into the infected mice . Similar increased production of TNF was also observed in mice treated with a nonlethal combination of heat-killed C . albicans and LPS . The number of viable C . albicans in kidneys of the infected mice was increased by LPS treatment, which was assumed to be the main cause of the greater mortality rate.

J Bacteriol, 1995 Jun, 177(11), 2971 - 6
D-arabitol metabolism in Candida albicans: construction and analysis of mutants lacking D-arabitol dehydrogenase; Wong B et al.; Candida albicans produces large amounts of the acyclic pentitol D-arabitol in culture and in infected animals and humans, and most strains also grow on minimal D-arabitol medium . An earlier study showed that the major metabolic precursor of D-arabitol in C . albicans was D-ribulose-5-PO4 from the pentose pathway, that C . albicans contained an NAD-dependent D-arabitol dehydrogenase (ArDH), and that the ArDH structural gene (ARD) encoded a 31-kDa short-chain dehydrogenase that catalyzed the reaction D-arabitol + NAD <=> D-ribulose + NADH . In the present study, we disrupted both ARD chromosomal alleles in C . albicans and analyzed the resulting mutants . The ard null mutation was verified by Southern hybridization, and the null mutant's inability to produce ArDH was verified by Western immunoblotting . The ard null mutant grew well on minimal glucose medium, but it was unable to grow on minimal D-arabitol or D-arabinose medium . Thus, ArDH catalyzes the first step in D-arabitol utilization and a necessary intermediate step in D-arabinose utilization . Unexpectedly, the ard null mutant synthesized D-arabitol from glucose . Moreover, 13C nuclear magnetic resonance studies showed that the ard null mutant and its wild-type parent synthesized D-arabitol via the same pathway . These results imply that C . albicans synthesizes and utilizes D-arabitol via separate metabolic pathways, which was not previously suspected for fungi.

Infect Immun, 1995 Jun, 63(6), 2378 - 81
Beta-1,2-linked oligomannosides from Candida albicans act as signals for tumor necrosis factor alpha production; Jouault T et al.; Different cell wall components from Candida albicans have been shown to stimulate murine macrophages for tumor necrosis factor alpha (TNF-alpha) secretion . All of these molecules contain beta-1,2-oligomannosides . In order to examine their role in TNF-alpha production, acid-labile oligosaccharides, released from C . albicans VW32 cell wall phosphopeptidomannan by mild acid hydrolysis, and previously shown to correspond to homopolymers of beta-1,2-linked mannopyranosyl units, were separated by gel filtration chromatography according to their degree of polymerization . Murine macrophages incubated with purified oligomannosides (M2 to M8) released TNF-alpha to an extent which was dependent on, although not directly correlated with, the length of the mannosyl chain . Slight activity was observed with M4 and M5; M6 and M7 had virtually no effect, whereas M8 was associated with strong TNF-alpha release . This effect of M8 was dose dependent and was not altered by polymyxin B, known to interfere with lipopolysaccharide-induced TNF-alpha production . These results suggest that stimulation of TNF-alpha release by C . albicans glycoconjugates containing beta-1,2-linked oligomannosides may be due, at least in part, to the presence of these components.

Infect Immun, 1995 Jun, 63(6), 2173 - 9
Evidence for the presence of collagenous domains in Candida albicans cell surface proteins; Sepulveda P et al.; Rabbit polyclonal antibodies (PAbs) directed towards the amino-terminal cysteine-rich 7S domain (PAb anti-7S), the major internal collagenous domain (PAb anti-type IV), and the C-terminal noncollagenous region (PAb anti-NC1) of the type IV collagen molecule were probed by indirect immunofluorescence against Candida albicans blastoconidia and germinated blastoconidia . Most nongerminating cells and mother blastoconidia from which germ tubes originated showed strong fluorescence when PAb anti-7S was used, whereas with PAb anti-type IV, fluorescence was found almost exclusively on the surface of filamentous forms . A patched fluorescent pattern rather than a homogenous confluent fluorescence was observed in all cases . No fluorescent cells were observed with PAb anti-NC1 . By Western immunoblotting, PAb anti-type IV cross-reacted primarily with a polypeptide of 37 kDa present in wall extracts obtained from intact cells of both growth forms by treatment with beta-mercaptoethanol, whereas PAb anti-7S recognized a major 58-kDa antigen also present in both extracts, along with some other high-molecular-mass (> 106-kDa) polydisperse species present only in the material released from blastoconidia with beta-mercaptoethanol . No reactive bands were observed when PAb anti-NC1 was used as a probe in Western immunoblotting experiments . The sensitivities or resistances to collagenase digestion of the different polypeptides that cross-reacted with PAbs anti-type IV and anti-7S suggest the existence of cell wall components in C . albicans that contain epitopes that mimic the collagenous domains of the type IV collagen molecule.

Infect Immun, 1995 Jun, 63(6), 2126 - 32
Comparative study of the C3d receptor and 58-kilodalton fibrinogen-binding mannoproteins of Candida albicans; Lopez-Ribot JL et al.; Using polyclonal antibodies (PAbs) raised against the Candida albicans C3d receptor (CR2; PAb anti-CR2) and the 58-kDa fibrinogen-binding mannoprotein (mp58; PAb anti-mp58) as well as ligand interactions, we have studied the relationship between these two receptors . In an indirect immunofluorescence assay with germ tubes, greater intensity was observed on the mother blastoconidium when PAb anti-CR2 was used, whereas greater intensity was localized to the hyphal extension when PAb anti-mp58 or binding of soluble fibrinogen was used . No competition or change in the fluorescence pattern was observed in dual-labeling experiments with PAb anti-CR2 and either fibrinogen or PAb anti-mp58 . Binding competition also was not observed in an enzyme-linked immunosorbent assay using the components present in a beta-mercaptoethanol extract from the cell wall of germ tubes . In immunoblots, PAb anti-CR2 recognized three different discrete bands with apparent molecular masses of 21, 40, and 66 kDa in the beta-mercaptoethanol extracts from the cell wall, whereas a different, single, broader band with an apparent molecular mass of 58 kDa was detected with PAb anti-mp58 . However, when nondenaturing conditions were used to separate the materials present in the cell wall extracts, no reactivity could be detected on Western blots (immunoblots) with PAb anti-mp58 . When PAb anti-CR2 was used for analysis, a single band migrating in the area corresponding to approximately 40 kDa was detected . These observations suggest a higher molecular weight for mp58 and one or more of the components detected with PAb anti-CR2 in their native state.

Arch Oral Biol, 1995 Jun, 40(6), 581 - 4
Modulation of the anti-Candida activity of apo-lactoferrin by dietary sucrose and tunicamycin in vitro; Nikawa H et al.; The sensitivity to apo-lactoferrin (apo-LF) of two oral isolates each of Candida albicans and C . krusei preincubated in either sucrose- or tunicamycin-supplemented media was investigated . Known apo-LF-sensitive isolates of both C . albicans and C . krusei demonstrated significantly increased resistance to apo-LF when preincubated in sucrose-supplemented media, but such preincubation had no effect on an apo-LF-resistant isolate of C . albicans, irrespective of the sugar concentration in the medium . Surprisingly, an apo-LF-resistant isolate of C . krusei demonstrated increased susceptibility to apo-LF when preincubated in sucrose . Further, when preincubated with tunicamycin, a chemical that interferes with candidal cell-wall formation, the susceptibility to apo-LF of the resistant isolates increased significantly . These results, when extrapolated in vivo, imply that dietary sucrose may diminish the anticandidal potency of salivary lactoferrin . However, the aberrant behaviour of a single isolate of C . krusei under these circumstances is possibly due to the different cell-wall structure of this yeast and needs to be further investigated.

Arch Oral Biol, 1995 Jun, 40(6), 577 - 9
Oral Candida albicans biotypes in Chinese patients with and without oral candidosis; Xu YY et al.; A total of 53 oral Candida albicans isolates from Chinese patients with clinically diagnosed oral candidosis (27 patients) or without overt signs and mycological manifestations of infection (26) were biotyped using two commercially available API micromethod kits and a boric acid-resistance test . There were no significant differences in the biotypes in health and disease, although the biotype A1R was present only in diseased individuals . The biotype A1S accounted for 21% of the total isolates, as in a number of other previous studies from the West . However, 14 of the 27 biotypes characterized were new biotypes that have not been described before . These preliminary data indicate that biotypic profile of C . albicans may bear no relation to the virulence of the isolates, and that diverse subtypes of the fungus are globally prevalent.

Microbiology, 1995 Jun, 141 ( Pt 6), 1301 - 8
Neocallimastix frontalis enolase gene, enol: first report of an intron in an anaerobic fungus; Durand R et al.; A DNA clone containing a putative enolase gene was isolated from a genomic DNA library of the anaerobic fungus Neocallimastix frontalis . It was deduced from sequence comparisons that the enolase gene was interrupted by a large 331 bp intron . The enolase gene, termed enol, has an ORF of 1308 bp and encodes a predicted 436 amino acid protein . The deduced amino acid sequence shows high identity (71.5-71%) to those of enolases from the yeasts Saccharomyces cerevisiae and Candida albicans . The G+C content of the enolase coding sequence (43.8 mol%) is considerably higher than the G+C content of the intervening sequence (14.2 mol%) or the 5' and 3' non-translated flanking sequences (15.2 and 4.7 mol%, respectively) . The codon usage of the N . frontalis enolase gene was very biased as has been found for the highly expressed genes of yeast and filamentous fungi . The gene has all the canonical features (polyadenylation signal, intron splicing boundaries) of genes isolated from aerobic filamentous fungi . Only one enolase gene could be detected in N . frontalis genomic DNA by Southern analysis with a homologous probe . RNA analysis detected a single enolase transcript of about 1.6 kb . When mycelium was grown on glucose, levels of enolase mRNA were markedly increased by comparison with enolase mRNA levels in mycelium grown on cellulose, suggesting that expression of the N . frontalis enolase gene was transcriptionally regulated by the carbon source.

Cytometry, 1995 Jun 1, 20(2), 127 - 33
Rapid flow cytometric method for measuring lymphocyte subset activation; Maino VC et al.; Standard in vitro methods for assessing T-cell activation have typically measured either the proliferative responses of peripheral blood mononuclear cell (PBMC) cultures to various provocative stimuli employing tritiated thymidine incorporation or the secretion of specific cytokines from activated cells . These bulk assay methods suffer the drawback of being lengthy assays, and, in addition, they do not provide information about functional responses of individual lymphocyte subsets . This report describes a three-color flow cytometric method for the rapid analysis (4 hours) of individual T-cell subsets in whole blood responding to various provocative stimuli, including pokeweed mitogen, the comitogenic monoclonal antibodies (mAbs) CD2/CD2R, the superantigen staphylococcal enterotoxin B (SEB), and the specific antigen Candida albicans . After 4 hours, CD69 expression in response to CD2/CD2R paralleled thymidine incorporation measured after 72 hours . Variations in the proportions of CD4+ and CD4- T cells expressing CD69 were observed with different stimuli . These observations demonstrate the potential of multiparameter flow cytometry for the investigation of functional responses of individual T-cell subsets to a variety of stimuli in whole blood.

J Clin Microbiol, 1995 Jun, 33(6), 1501 - 9
Colonizing populations of Candida albicans are clonal in origin but undergo microevolution through C1 fragment reorganization as demonstrated by DNA fingerprinting and C1 sequencing; Lockhart SR et al.; The genetic homogeneity of nine commensal and infecting populations of Candida albicans has been assessed by fingerprinting multiple isolates from each population by Southern blot hybridization first with the Ca3 probe and then with the 0.98-kb C1 fragment of the Ca3 probe . The isolates from each population were highly related, demonstrating the clonal origin of each population, but each population contained minor variants, demonstrating microevolution . Variation in each case was limited to bands of the Ca3 fingerprint pattern which hybridized with the 0.98-kb C1 fragment . The C1 fragment was therefore sequenced and demonstrated to contain an RPS repetitive element . The C1 fragment also contained part or all of a true end of the RPS element . These results, therefore, demonstrate that most colonizing C . albicans populations in nonimmuno-suppressed patients are clonal, that microevolution can be detected in every colonizing population by C1 hybridization, and that C1 contains the repeat RPS element.

Clin Exp Allergy, 1995 Jun, 25(6), 522 - 8
Detection of IgE antibody against Candida albicans enolase and its crossreactivity to Saccharomyces cerevisiae enolase; Ito K et al.; Candida albicans 46 kDa protein, a glycolytic enolase enzyme, is an important allergen of the yeast . The purpose of the study was to detect circulating IgE and IgG antibodies against C . albicans enolase (CAE) . We isolated CAE using sequential DEAE Sephacel and P11 column chromatography from spheroplasts of C . albicans, and detected IgE and IgG antibody against CAE by immunoblotting . Crossreactivity of enolase of C . albicans and Saccharomyces cerevisiae was also examined by immunoblotting and immunoblot inhibition test . Among 54 sera with positive IgE RAST to C . albicans, IgE antibody against CAE was detected in 20 sera (37%) and IgG antibody in 27 sera (50%) . The allergenic potency of CAE was confirmed using a skin-prick test in three patients . Simultaneous IgE binding to S . cerevisiae enolase was only observed in four out of 20 sera reacting to CAE . Pre-treatment of sera with CAE completely inhibited IgE binding to S . cerevisiae enolase . Whereas the latter only partially inhibited IgE binding to CAE . These results suggest that CAE shares some crossreacting epitopes with S . cerevisiae enolase, representing minor components of CAE but dominant segments of S . cerevisiae enolase.

Trends Microbiol, 1995 Jun, 3(6), 237 - 40
A TH1-TH2-like switch in candidiasis: new perspectives for therapy; Puccetti P et al.; An imbalance in TH1-type and TH2-type responses may allow Candida albicans to modify the host response to favor its own persistence . This hypothesis has important consequences for allergy, autoimmunity and co-infection, and also highlights a potential role for cytokine and anti-cytokine therapy in Candida-related pathology.

Eur J Immunol, 1995 Jun, 25(6), 1559 - 65
Interleukin-4 and -10 exacerbate candidiasis in mice; Tonnetti L et al.; Neutralization of endogenous interleukin (IL)-4 or IL-10 in mice with Candida albicans infection initiates or accelerates development of a T helper (Th)1-associated protective response . Here, we report the effect of IL-4 and IL-10 administration on the course of systemic or gastrointestinal (GI) candidiasis and on the development of Th immunity using yeast/host combinations that result either in Th1-associated self-limiting infection (healer mice) or in Th2-associated progressive disease (nonhealer mice) . Treatment with IL-4 or IL-10 greatly exacerbated the course of systemic infection in nonhealer mice and rendered healer mice, inoculated with attenuated yeast cells, susceptible to infection . Under the latter conditions of yeast challenge and IL-4/IL-10 administration, the development of a fatal disease was associated with inhibition of IL-12 production and detection of progressive Th2 cell dominance . In contrast, in healer mice allowed to resolve their infections and to develop long-lived anti-candidal resistance, the expression of this acquired resistance was not impaired by IL-4 and/or IL-10, as shown by the outcome of reinfection with virulent yeast cells . In the GI model of infection, both IL-4 and IL-10 were found to exacerbate the course of infection and to induce the appearance of CD4+ T cells producing high levels of IL-4 and IL-10 in Peyer's patches . These findings demonstrate that exogenous IL-4 and IL-10 may greatly affect the development of Th responses to C . albicans in vivo, but do not modify the expression of established and predominant Th1 cell reactivity.

Exp Mycol, 1995 Jun, 19(2), 101 - 10
Detection of myosin immunoanalogue in the yeast Candida albicans; Ghazali M et al.; Detection and localization of myosin immunoanalogue protein in the yeast Candida albicans were achieved by immunoblotting, indirect immunofluorescence assay, and immunoelectron microscopy . A polypeptide with an M(r) about 110,000, from cytosolic extract and insoluble fraction in the corresponding membrane pellet, was reacted with polyclonal and monoclonal antibodies raised against vertebrate muscle myosin . This protein was located by immunofluorescence and immunoelectron microscopy in the cell cortex along the plasmalemma, in the cytoplasm, and in the septum corresponding to bud scar region situated between the yeast-mother cell and the bud.

Nat Med, 1995 Jun, 1(6), 552 - 7
Gamma delta T cell-induced nitric oxide production enhances resistance to mucosal candidiasis; Jones-Carson J et al.; Despite the prevalence of gamma delta T cells in mucosae that are typically colonized by Candida albicans, little is known of the possible role of these cells in resistance to candidiasis . A sharp increase in the number of gamma delta T cells and macrophages following intraperitoneal inoculation of mice with C . albicans led us to examine the role of these cells in the immune response to C . albicans . We show that the gamma delta T cells enhance macrophage nitric oxide (NO) production and anti-candida activity, in vitro . We also propose that the gamma delta T cells regulate macrophage function during candidiasis in vivo as well, because depletion of these cells abrogated inducible NO synthase expression in mucosae and enhanced murine susceptibility to candidiasis.

FEMS Immunol Med Microbiol, 1995 Jun, 11(3), 157 - 62
Nitric oxide-dependent killing of Candida albicans by murine peritoneal cells during an experimental infection; Rementeria A et al.; The phagocytic and candidacidal activities of the peritoneal cells of Candida albicans-infected mice were studied 20 days following experimental infection . Both activities were enhanced during infection . The production of nitric oxide (NO) by the peritoneal cells of infected mice was determined, and an increase in the nitrite concentration in supernatants of peritoneal cell cultures was detected . The period of NO production by the peritoneal cells coincided partially with the period of enhanced C . albicans killing . The inhibition of NO synthesis by N-monomethyl-L-arginine was concomitant with inhibition of candidacidal activity . We conclude that NO synthesis is the primary candidacidal mechanism of the murine peritoneal cells activated by C . albicans infection.

J Hosp Infect, 1995 Jun, 30 Suppl, 209 - 17
Infections in liver transplantation: risk factors and strategies for prevention; Kibbler CC; Infection affects up to 70% of liver transplant recipients and is the second most common complication after rejection and graft dysfunction . Identified risk factors for infection include: previous transplantation; type of biliary anastomosis; transfusion requirements at surgery; surgical complications; duration of operation; duration of postoperative ventilation; serological status of donor and recipient; steroid use and serotherapy for rejection; and pre- and post-transplant antibiotic usage . The majority of symptomatic infections are bacterial and relate to surgery (intra-abdominal, biliary and wound infections), ventilation and intravenous cannulae . Cytomegalovirus infections occur in 45-100% of recipients but are asymptomatic in the majority . Fungal infections are mostly due to Candida albicans but infections due to Aspergillus spp . occur in approximately 6% and carry a high mortality . There are very few prospective comparative trials of antimicrobial prophylaxis in this patient population . The management of these patients needs to be based on such studies.

J Antimicrob Chemother, 1995 Jun, 35(6), 793 - 804
Correlation of in-vitro susceptibility test results with clinical response: a study of azole therapy in AIDS patients; Rodriguez-Tudela JL et al.; The in-vitro susceptibilities of 40 clinical isolates of Candida albicans to ketoconazole and fluconazole were determined and an attempt was made to correlate these data with the clinical responses of the patients from whom the strains were originally isolated to treatment with these agents . Of 40 patients with the acquired immunodeficiency syndrome (AIDS) with oropharyngeal and/or oesophageal candidosis, 21 received ketoconazole and 19 fluconazole . Susceptibility testing was performed by a microbroth dilution method with RPMI-2% glucose medium according to the recommendations of the National Committee for Clinical Laboratory Standards; growth inhibition was estimated spectrophotometrically and the MIC endpoint was defined in terms of the IC1/2 . The MICs of 236 additional strains of C . albicans, which were also isolated from AIDS patients, were used to establish a susceptibility profile for this species . On the basis of the susceptibility test results and the clinical responses of the 40 patients, the following tentative breakpoints for ketoconazole and fluconazole are proposed: patients with infections caused by C . albicans strains with MICs of ketoconazole and fluconazole or < or = 0.001 and < or = 0.25 mg/L respectively would be expected to respond to treatment with these agents and isolates with MICs which meet these criteria are therefore classified as susceptible; patients with infections caused by strains with MICs of ketoconazole and fluconazole of > or = 0.06 and > or = 16.0 mg/L respectively would not be expected to respond to treatment with these agents and isolates with MICs which meet these criteria are therefore classified as resistant; the response of patients with infections caused by strains with MICs of ketoconazole and fluconazole of 0.003-0.03 and 0.5-8.0 mg/L respectively cannot be reliably predicted and isolates with MICs which fall within these ranges are therefore classified as being of indeterminate susceptibility . The present study demonstrates that the results of in-vitro susceptibility testing with RPMI-2% glucose broth correlate with the clinical response to therapy and can be used to facilitate optimal treatment in AIDS patients with oropharyngeal and/or oesophageal candidosis.

J Antimicrob Chemother, 1995 Jun, 35(6), 739 - 49
Defining conditions for microbroth antifungal susceptibility tests: influence of RPMI and RPMI-2% glucose on the selection of endpoint criteria; Rodriguez-Tudela JL et al.; We have compared amphotericin B, flucytosine, ketoconazole and fluconazole susceptibilities of 40 clinical isolates of Candida albicans by broth microdilution in two different media: RPMI 1640 (RPMI) and the same medium supplemented with 18 g of glucose per L (RPMI-2% glucose) . Preparation of media, drugs and inocula, as well as incubation temperature, followed the recommendations of the National Committee for Clinical Laboratory Standards (Villanova, PA, USA) antifungal agent working group for broth macrodilution tests with antifungal agents . Microtitre plates were agitated for 5 min before spectrophotometric readings were performed with an automatic plate reader . MIC endpoints were defined in three different ways: (i) MIC-100% for amphotericin B and flucytosine; (ii) MIC-80% for azole-drugs and also for flucytosine; (iii) IC1/2 for azole-drugs . The MIC endpoints were compared between the two different media in order to ascertain which was the best criterion . For amphotericin B, 100% of strains had a maximum difference of a twofold dilution in both media . For flucytosine, MIC values were very similar in both media, irrespective of the MIC endpoint chosen, MIC-100% or MIC-80% . For azole-drugs the (MIC-80%)50 and (MIC-80%)90 in RPMI were higher than those in RPMI-2% glucose . However, (IC1/2)50 and (IC1/2)90 were identical in both media as well as (MIC-80%)50 and (MIC-80%)90 in RPMI-2% glucose, The limited growth of yeasts in RPMI precludes the selection of an azole-MIC-80% endpoint, although the MIC determination was performed with an objective turbidimetric method (spectrophotometric reading plus mathematical MIC calculation) . The use of RPMI-2% glucose produces the same MIC whatever methods was used, IC1/2 or MIC-80% . However, some minor discrepancies can be expected between IC1/2 and MIC-80% when strains with higher "trailing effect" are being tested . Therefore, we recommended IC1/2 in RPMI-2% glucose as the method of choice for MIC calculation, until more studies correlating in-vitro results with clinical outcome have been performed.

Biol Pharm Bull, 1995 Jun, 18(6), 923 - 5
Bioactivity of nickel(II) complex containing N-glycosides derived from D-glucosamine and ethylenediamine against pathogenic yeast, Candida albicans; Yano S et al.; A nickel(II) complex containing N-glycosides derived from the reaction of D-glucosamine (D-GlcN) and ethylenediamine (en) {Ni(D-GlcN-en)2}Cl2.H20 showed effective antifungal activity against pathogenic yeast, Candida albicans, where the MIC (minimal concentration of inhibition) value of the complex is 0.25 mM.

Clin Exp Allergy, 1995 Jun, 25(6), 529 - 35
Characterization of IgE-binding epitopes on Candida albicans enolase; Ito K et al.; Candida albicans enolase is one of the important allergens in Candida allergy . We isolated and purified 46kDa C . albicans enolase (CAE) from C . albicans and characterized epitopes for IgE antibody by lectin-blotting and enzymatic digestion followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting . Lectin blotting and deglycozilation indicated that this protein did not contain polysaccharide side chains . The purified CAE and recombinant fusion protein produced from CAE gene possessed common epitopes for IgE antibody . We estimated IgE binding epitopes on the basis of reported amino acid sequences from the analysis of cDNA encoding CAE . V8 protease digestion of CAE gave six polypeptide fragments (A-F) . The N-termini of each fragment were confirmed by amino acid sequence and the C-termini were estimated by molecular weights of each fragment and the specific cutting site of V8 protease . Fragment C (25.0 kDa; F-171-I-399) reacted to 90% IgE antibodies examined, whereas fragments D (21.0 kDa; F-171-I-360), E (16.2 kDa; F-171-D-317) and F (13.0 kDa; A-47-E-170) showed no IgE binding . Our results suggest that epitopes for IgE antibodies exist near the C-terminal of the protein.

J Infect Dis, 1995 Jun, 171(6), 1668 - 71
Preliminary assessment of a human recombinant antibody fragment to hsp90 in murine invasive candidiasis; Matthews R et al.; Seroconversion to hsp90 is associated with recovery from systemic candidiasis in humans, and a murine monoclonal antibody to this hsp90 antigen (LKVIRK epitope) was protective in mice . A human recombinant antibody to the same epitope was assessed in acute and chronic models of murine invasive candidiasis . Lethal intravenous challenge with fluconazole-susceptible (strain 4) or fluconazole-resistant (strain 019) Candida albicans, followed 2 h later by a single dose of recombinant antibody, was associated with a statistically significant drop in mortality of > or = 40% (two experiments in BALB/c mice given strain 4; one experiment in CD-1 mice given strain 019) or 23% (BALB/c mice, strain 019) . In mice sublethally infected with strain 4, treatment with recombinant antibody was associated with improved renal clearance of infection . Antibody-mediated protection may involve neutralization of the protein-binding properties of circulating candidal hsp90, since LKVIRK strongly bound dexamethasone in vitro.

Med Hypotheses, 1995 Jun, 44(6), 507 - 15
Chronic intestinal candidiasis as a possible etiological factor in the chronic fatigue syndrome; Cater RE 2nd; The chronic candidiasis syndrome, also known as the Candida-related complex, putatively caused by the overgrowth of Candida albicans in the gastrointestinal tract and secondarily in the genital organs, is briefly described . Patients with this disorder have many of the same symptoms as those with the chronic fatigue syndrome, except for the recurrent flu-like symptoms of the latter disorder . The positive response of a large number of patients with the chronic fatigue syndrome (CFS) to an oral antifungal agent and a diet for intestinal candidiasis has been described by another clinician . There is evidence that Candida albicans infection of the mucous membranes depresses T cell and natural killer (NK) cell function . Similar abnormalities of immune function are found in the CFS . The function of cytotoxic T cells, T helper cells, and NK cells is important in preventing reactivation of infections from Epstein-Barr virus, cytomegalovirus, and other herpesviruses . Reactivation of one or more of these viruses could lead to the expression of the flu-like symptoms in the CFS . Yet the immune dysfunction found in this disorder has been considered the primary underlying causal factor . It is proposed that chronic intestinal candidiasis may be an agent which leads to immune depression in many CFS patients and therefore that it could be a causal factor in CFS.

FEMS Microbiol Lett, 1995 May 15, 128(3), 271 - 7
Glucosylation of cell wall proteins in regenerating spheroplasts of Candida albicans; Kapteyn JC et al.; The cell wall of Candida albicans contains mannoproteins that are covalently associated with beta-1,6-glucan . When spheroplasts were allowed to regenerate a new cell wall, initially non-glucosylated cell wall proteins accumulated in the medium . While the spheroplasts became osmotically stable, beta-1,6-glucosylated proteins could be identified in their cell wall by SDS-extraction or beta-1,3-glucanase digestion . At later stages of regeneration, beta-1,3-glucosylated proteins were also found . Hence, incorporation of proteins into the cell wall is accompanied by extracellular coupling to beta-1,6-/beta-1,3-glucan . The SDS-extractable glucosylated proteins probably represent degradation products of wall proteins rather than their precursors . Tunicamycin delayed, but did not prevent the formation of beta-1,6-glucosylated proteins, demonstrating that beta-1,6-glucan is not attached to N-glycosidic side-chains of wall proteins.

J Immunol, 1995 May 15, 154(10), 5273 - 81
Growth inhibition of Candida albicans hyphae by CD8+ lymphocytes; Beno DW et al.; We have shown previously that IL-2-activated splenocytes can inhibit the growth of Candida albicans hyphae in vitro . Herein we demonstrate that plastic nonadherent lymphocytes that are CD8+ mediate the antifungal activity . Enrichment for CD8+ cells markedly enhanced the antifungal activity of the IL-2-activated lymphocyte population for C . albicans and the cytotoxic activity of the lymphocytes for an NK-resistant cell line . Depletion of CD8+ cells reduced the lymphocyte population's antifungal activity and cytotoxic activity for the NK-resistant cell line . Enrichment for NK1.1+ cells markedly reduced the antifungal activity of the lymphocyte population for C . albicans and increased the cytotoxic activity of the lymphocytes for an NK-sensitive cell line . Depletion of NK1.1+ cells increased the lymphocyte population's antifungal activity and cytotoxic activity for the NK-resistant cell line . Generation of the antifungal lymphocytes in culture required IL-2 and was not replaced with IFN-gamma . These data show that IL-2-activated CD8+ T lymphocytes exert the greatest amount of antifungal effect against the hyphal form of C . albicans, whereas IL-2- or IFN-gamma-activated NK cells have little or no effect against the hyphae.

Eur J Biochem, 1995 May 15, 230(1), 111 - 8
Selenomethionine labelling of phosphomannose isomerase changes its kinetic properties; Bernard AR et al.; Phosphomannose isomerase (PMI) is an essential enzyme in the early steps of the protein glycosylation pathway in both prokaryotes and eukaryotes . Lack of the enzyme is lethal for fungal organisms and it is thus a potential fungicidal target . To facilitate the solution of the three-dimensional structure of the enzyme from the pathogen Candida albicans, we have produced the recombinant selenomethionine-labelled enzyme (SeMet-PMI) . DL41, a methionine auxotroph Escherichia coli strain, was transformed with a PMI expression plasmid and grown on an enriched selenomethionine-containing medium to high-cell densities . The SeMet-PMI protein has been purified and found by amino acid analysis to have its methionine residues replaced by selenomethionine residues . Electrospray mass spectroscopy showed a major species of 49,063 +/- 10 Da for SeMet-PMI compared to 48,735 +/- 6 Da for the normal recombinant enzyme, accounting for the incorporation of seven selenomethionine residues . SeMet-PMI crystallised isomorphously with the normal PMI protein and the crystals diffract to 0.23 nm . Kinetic characterisation of SeMet-PMI showed that its Km for the substrate mannose-6-phosphate was fourfold higher than that of its methionine-containing counterpart . The inhibition constant for zinc ions was also increased by a similar factor . However, the Vmax was unaltered . These results suggested that one or more methionine residues must be in close proximity to the substrate-binding pocket in the active site, rendering substrate access more difficult compared to the normal enzyme . This hypothesis was confirmed by the finding of four methionine residues lying along one wall of the active site.

Nucleic Acids Res, 1995 May 11, 23(9), 1481 - 6
The CUG codon is decoded in vivo as serine and not leucine in Candida albicans; Santos MA et al.; Previous studies have shown that the yeast Candida albicans encodes a unique seryl-tRNA(CAG) that should decode the leucine codon CUG as serine . However, in vitro translation of several different CUG-containing mRNAs in the presence of this unusual seryl-tRNA(CAG) result in an apparent increase in the molecular weight of the encoded polypeptides as judged by SDS-PAGE even though the molecular weight of serine is lower than that of leucine . A possible explanation for this altered electrophoretic mobility is that the CUG codon is decoded as modified serine in vitro . To elucidate the nature of CUG decoding in vivo, a reporter system based on the C . albicans gene (RBP1) encoding rapamycin-binding protein (RBP), coupled to the promoter of the C . albicans TEF3 gene, was utilized . Sequencing and mass-spectrometry analysis of the recombinant RBP expressed in C . albicans demonstrated that the CUG codon was decoded exclusively as serine while the related CUU codon was translated as leucine . A database search revealed that 32 out of the 65 C . albicans gene sequences available have CUG codons in their open reading frames . The CUG-containing genes do not belong to any particular gene family . Thus the amino acid specified by the CUG codon has been reassigned within the mRNAs of C . albicans . We argue here that this unique genetic code change in cellular mRNAs cannot be explained by the 'Codon Reassignment Theory'.

J Biochem (Tokyo), 1995 May, 117(5), 1131 - 7
A novel big defensin identified in horseshoe crab hemocytes: isolation, amino acid sequence, and antibacterial activity; Saito T et al.; Hemocytes of the horseshoe crab (limulus) contain a family of arthropodous peptide antibiotics, termed the tachyplesin family, and antibacterial protein, called anti-LPS factor, of which the former is located in the small (S) granules and the latter in the large (L) granules of the hemocytes . In our ongoing studies on granular components, we have identified here a novel defensin-like substance present in both L- and S-granules . This substance strongly inhibits the growth of Gram-negative and -positive bacteria, and fungi, such as Candida albicans . The isolated substance, tentatively termed "big defensin," consists of 79 amino acid residues, of which the COOH-terminal 37 residues have a sequence similar to those of mammalian neutrophil-derived defensins, especially rat defensin . Characterization of the disulfide motif in big defensin indicated that the disulfide array is identical to that of beta-defensins from bovine neutrophils . One clear structural difference is that the limulus hemocyte-derived big defensin has an extension of the NH2-terminal hydrophobic sequence with 35 amino acid residues followed by the COOH-terminal cationic defensin portion . This amphipathic nature of big defensin seems likely to be associated with its potent antibacterial activity . Furthermore, antibacterial activities of the NH2-terminal hydrophobic region and the COOH-terminal defensin portion separated by tryptic digestion are significantly different: the former displays a more potent activity against Gram-positive bacteria, whereas the latter is more potent against Gram-negative bacteria . Big defensin, therefore, may prove to represent a new class of defensin family possessing two functional domains with different antimicrobial activities.

Gen Diagn Pathol, 1995 May, 141(1), 35 - 9
Neuropathologic findings after organ transplantation . An autopsy study; Schwechheimer K et al.; Since 1972 organ transplantations of kidney, bone marrow, liver, heart and lung have been performed at the University Hospital of Essen, Germany . Out of 2535 transplantations until September 1993, autopsies were performed in 157 patients In 25 patients (15.9%) neuropathologic findings (n = 26) were found . In 97 autopsies after bone marrow transplantation, 9 patients (9.3%) exhibited a severe neuropathologic alteration . In six patients (6/9; 66.6%), necrotisizing toxoplasmose encephalitis was found . Other cases showed a septic-metastatic mycotic encephalitis with crypto-coccus neoformans and candida albicans (n = 2) and leucemia infiltrates (n = 1) . Massive cerebral hemorrhage was the most frequent neuropathologic finding after liver (4/8) and kidney transplantation (3/6) . In addition liver-transplanted patients exhibited septic-metastatic encephalitis (3/8) and embolic brain infarct (1/8) as well as cerebral metastases (2/6) and primary malignant cerebral lymphoma in kidney transplantation (1/6) . CNS findings in five autopsies after heart-lung-transplantation were diverse . They comprised intracerebral hemorrhage, intravasal lymphoma and septic-metastatic encephalitis, respectively . In summary, neuropathologic autopsy findings after organ transplantation are diverse and preferentially comprise infections, cerebral hemorrhages, and malignant lymphomas . After bone marrow transplantation, the most frequent neuropathologic autopsy finding was toxoplasmose encephalitis and massive cerebral hemorrhages after liver and kidney transplantations.

Mycoses, 1995 May-Jun, 38(5-6), 235 - 7
The aetiological agents of superficial cutaneous mycoses in Jos, Plateau State of Nigeria; Ayanbimpe GH et al.; A survey of superficial skin mycoses was carried out among miners and office workers employed in different establishments in Jos, Nigeria . Mycotic infection was demonstrable by microscopy and culture in 45 (10.4%) subjects: 20 males and 25 females . Malassezia furfur was the predominant aetiological agent, followed by Candida albicans and Trichophyton soudanense . Other aetiological agents frequently recovered were T . rubrum., T . mentagrophytes., Microsporum audouinii and Trichosporon beigelii.

Mycoses, 1995 May-Jun, 38(5-6), 191 - 5
Preliminary studies of the antifungal activities of some medicinal plants against Basidiobolus and some other pathogenic fungi; Nwosu MO et al.; The antifungal activities of extracts of 10 medicinal plants collected from south-eastern parts of Nigeria were tested against seven pathogenic fungi using the broth dilution and agar plate methods . All the extracts at 1:10 dilution inhibited the growth of Basidiobolus haptosporus and B . ranarum but did not inhibit that of Aspergillus fumigatus, Geotrichum candidum and Candida albicans . While extracts from Piper guineense, Ocimum gratissimum, Moringa oleifera and Erythrophleum suaveolens inhibited the growth of Trichophyton rubrum and T . mentagrophytes, those from Fatropha curcas, Mitracarpus villosus, Azadirachta indica and Gongronema latifolium failed to do so at 1:10 dilution . Extract from Piper sp . was also able to inhibit the growth of B . haptosporus at a concentration as low as 1:80 dilution followed by those of Ocimum and Rauvolfia spp . at 1:40 dilution . These results indicate possible use of certain plant extracts in the treatment of subcutaneous phycomycosis in humans and animals.

Neurosurgery, 1995 May, 36(5), 1009 - 12; discussion 1012-3
Candidal pituitary abscess: case report; Heary RF et al.; We report a case of a culture-proven intrasellar Candida albicans abscess . A 36-year-old woman presented with a history of headaches, menstrual irregularities, and mild symptoms of diabetes insipidus . She was neurologically intact at the time of a transsphenoidal surgery for a presumed pituitary adenoma . An extensive work-up revealed that although the patient was seronegative for human immunodeficiency virus, she was immunocompromised with a T-cell dysfunction . Fungal abscesses of the pituitary gland have rarely been reported . This is the first documented case of a patient who is seronegative for human immunodeficiency virus who becomes infected by an ordinarily innocuous fungus, Candida albicans.

J Dent Res, 1995 May, 74(5), 1152 - 61
Oral Candida: clearance, colonization, or candidiasis?
Cannon RD, Holmes AR, Mason AB, Monk BC.
Candida albicans is frequently isolated from the human mouth, yet few carriers develop clinical signs of candidiasis . Oral candidiasis presents clinically in many forms . This reflects the ability of the yeast to colonize different oral surfaces and the variety of factors which predispose the host to Candida colonization and subsequent infection . Colonization of the oral cavity appears to be facilitated by several specific adherence interactions between C . albicans and oral surfaces which enable the yeast to resist host clearance mechanisms . Thus, Candida has been shown to adhere to complement receptors, various extracellular matrix proteins, and specific sugar residues displayed on host or bacterial surfaces in the oral cavity . Oral candidiasis results from yeast overgrowth and penetration of the oral tissues when the host's physical and immunological defenses have been undermined . Tissue invasion may be assisted by secreted hydrolytic enzymes, hyphal formation, and contact sensing . While these and other phenotypic characteristics may endow certain Candida species or strains with a competitive advantage in the oral cavity, it is the host's immune competence that ultimately determines whether clearance, colonization, or candidiasis occurs.

Microbiology, 1995 May, 141 ( Pt 5), 1147 - 56
Cloning of a Candida albicans peptide transport gene; Basrai MA et al.; A Candida albicans peptide transport gene, CaPTR2, was cloned from a C . albicans genomic library by functional complementation of a peptide transport deficient mutant (strain ptr2-2) of Saccharomyces cerevisiae . CaPTR2 restored peptide transport to transformants as determined by uptake of radiolabelled dileucine, growth on dipeptides as sources of required amino acids, and restoration of growth inhibition by toxic peptides . Plasmid curing experiments demonstrated that the peptide transport phenotype was plasmid borne . CaPTR2 was localized to chromosome R of C . albicans by contour-clamped homologous electric field gel chromosome blots . Deletion subclones and frameshift mutagenesis were used to narrow the peptide transport complementing region to a 5.1 kb DNA fragment . DNA sequencing of the complementing region identified an ORF of 1869 bp containing an 84 nucleotide intron . The deduced amino acid sequence predicts a protein of 70 kDa consisting of 623 amino acids with 12 hydrophobic segments . A high level of identity was found between the predicted protein and peptide transport proteins of S . cerevisiae and Arabidopsis thaliana . This study represents the first steps in the genetic characterization of peptide transport in C . albicans and initiates a molecular approach for the study of drug delivery against this pathogen.

J Toxicol Environ Health, 1995 May, 45(1), 75 - 82
Effects of exposure to NO2 or SO2 on bronchopulmonary reaction induced by Candida albicans in guinea pigs; Kitabatake M et al.; The effects of NO2 or SO2 on the bronchopulmonary reactions induced by Candida albicans in guinea pigs were evaluated . Thirty-six guinea pigs (3 groups of 12 animals each) were sensitized with intraperitoneal injection of 10 mg of C . albicans, given twice . Two groups of animals were exposed to about 5 ppm of NO2 or SO2 for 4 h/d, 5 d/wk; this exposure was conducted a total of 30 times during the study . The third group served as the control and was not exposed to these pollutants . Two weeks after the second sensitization, all the animals were subjected to inhalation exposure to C . albicans . For 42 h after the antigen challenge, the respiratory rates and expiration/inspiration ratios of the animals were automatically monitored . The number of animals showing tachypnea was significantly higher in the NO2 exposure group than in the control from 15 h after antigen challenge . In the SO2 exposure group, the number of animals showing prolonged expiration or prolonged inspiration, or both, was significantly higher than that in the control group, and the symptoms were observed from approximately 15 h after antigen challenge . These findings showed that delayed-type dyspneic symptoms in guinea pigs were increased by exposure to NO2 or SO2, although the symptoms and degree of dyspnea were different for the two gases.

J Med Microbiol, 1995 May, 42(5), 372 - 9
The genotypic relationship of Candida albicans strains isolated from the oral cavity of patients with denture stomatitis; Mathaba LT et al.; Fifty-seven isolates of Candida albicans were obtained from different sites within the oral cavities of 18 dental patients without AIDS or any malignancies . Eleven of the patients had oral candidosis associated with the wearing of dentures . The genotypic relationships of the individual isolates were determined by hybridisation of a C . albicans-specific moderately repetitive sequence, 27A, to EcoRI-digested C . albicans chromosomal DNA . From the DNA profiles, the isolates could be divided into 22 distinct genetic groups . In the majority of patients, a single unique strain of C . albicans appeared to dominate in the oral cavity . Re-infection following antifungal therapy was generally due to the re-emergence of the original infecting strain . The C . albicans strains isolated from dental plates did not form a distinct genetic group . These results suggest that denture stomatitis is due to the outgrowth of commensal strains of C . albicans.

J Infect Dis, 1995 May, 171(5), 1289 - 94
Resistance of zinc-supplemented Candida albicans cells to the growth inhibitory effect of calprotectin; Santhanagopalan V et al.; Calprotectin is a neutrophil cytoplasmic protein with significant microbistatic activity . This protein may compete for zinc, or the metal may inactivate a different microbistatic activity of the protein . To distinguish between these possibilities, the sensitivity to calprotectin was determined for zinc-supplemented Candida albicans cells . The latter had increased growth in cultures containing either human empyema fluid as a source of calprotectin or purified calprotectin itself . This increased growth did not appear to be due to leakage of zinc into the medium . In other experiments, empyema fluid supernatants did not suppress C . albicans growth across a dialysis membrane; however, other studies suggested that it is difficult to significantly suppress C . albicans growth by zinc depletion unless the depleting agent remains in the cultures . These results indicate that calprotectin inhibits C . albicans growth through competition for zinc.

J Bacteriol, 1995 May, 177(10), 2953 - 5
The "universal" leucine codon CTG in the secreted aspartyl proteinase 1 (SAP1) gene of Candida albicans encodes a serine in vivo; White TC et al.; A number of Candida species possess a tRNA(Ser)-like species that recognizes CTG codons that normally specify leucine (Leu) in the universal code of codon usage . Mass spectrometry and Edman sequencing of peptides from the secreted aspartyl proteinase isoenzyme (Sap1) demonstrate that positions specified by the CTG codon contain a nonmodified serine (Ser) in Candida albicans.

EMBO J, 1995 May 1, 14(9), 1932 - 41
Lymphoproliferative disorder and imbalanced T-helper response in C/EBP beta-deficient mice; Screpanti I et al.; C/EBP beta is considered a key element of interleukin-6 (IL-6) signalling as well as an important transcriptional regulator of the IL-6 gene itself . We describe here how mice lacking C/EBP beta develop a pathology similar to mice overexpressing IL-6 and nearly identical to multicentric Castleman's disease in human patients, with marked splenomegaly, peripheral lymphadenopathy and enhanced haemopoiesis . Humoral, innate and cellular immunity are also profoundly distorted, as shown by the defective activation of splenic macrophages, the strong impairement of IL-12 production, the increased susceptibility to Candida albicans infection and the altered T-helper function . Our data show that C/EBP beta is crucial for the correct functional regulation and homeostatic control of haemopoietic and lymphoid compartments.

Cell Immunol, 1995 May, 162(2), 256 - 64
Cytokine response to inactivated Candida albicans in mice; Rosati E et al.; Inactivated Candida albicans (CA) cells induce strong activation of natural cytotoxic effectors in mice . In the present study we examined the expression of cytokine genes involved in the immune response to CA . It has been reported that differential cytokine production by natural immune cells is important for regulating the development of specific TH response . Northern blot analysis was performed on peritoneal exudate cells (PEC) recovered from CD2F1 mice injected ip with five doses of CA (CA-5d, on Days -14, -10, -7, -3, 0 with respect to the in vitro assays at 2, 24, and 72 hr) or from mice injected ip with four doses of CA (CA-4d, on Days -14, -10, -7, -3 with respect to the in vitro assay on Day 0) . On Day 0, before the fifth CA injection, PEC expressed a high level of IL-2 and a low level of IL-1 beta mRNAs while genes coding for IL-4, IL-5, IL-6, IL-10, IL-12, TNF alpha, and IFN gamma were not expressed and there was a high level of NK activity . Two hours after CA-5d a high level of IFN gamma and a low level of IL-10 mRNAs were already evident, while IL-2 and much more IL-1 beta had greatly increased . IL-6, TNF alpha, and IL-2R alpha chain mRNAs were also detectable, whereas IL-4, IL-5, and IL-12 were not expressed . IL-12 mRNA was also absent in earlier stages of the CA sensitization . Both cellularity and NK activity of peritoneal exudate had increased with respect to Day 0 . At 24 hr whereas IL-2 mRNA remained high, both IL-1 beta and IFN gamma mRNAs expression had decreased . Expression of other cytokines was no longer detectable but NK activity remained high and a significant LAK activity was also induced . After 72 hr, while the IL-2 mRNA level and NK activity were still high the IL-1 beta mRNA expression had further decreased . These results indicate that CA induces a predominant production of IFN gamma and IL-2, cytokines involved in the development of TH1 response but it is unable to induce IL-12 . This secondary pathway, without IL-12 involvement in the development of TH1 response, is probably the result of the ability of IL-2, IL-1 beta, and TNF alpha to synergize in inducing IFN gamma synthesis by NK cells.

Infect Immun, 1995 May, 63(5), 1993 - 8
Evidence implicating phospholipase as a virulence factor of Candida albicans; Ibrahim AS et al.; Three different approaches were used to investigate the role of extracellular phospholipases in the pathogenicity of Candida albicans . First, we compared 11 blood isolates of this yeast with an equal number of commensal strains isolated from the oral cavities of healthy volunteers . Blood isolates produced significantly more extracellular phospholipase activity than the commensal strains did . Second, two clinical isolates of C . albicans that differed in their levels of virulence in a newborn mouse model were compared for their ability to secrete phospholipases . The invasive strain produced significantly more extracellular phospholipase activity than the noninvasive strain did . Third, nine blood isolates were characterized for their phospholipase and proteinase production, germ tube formation, growth, and adherence to and damage of endothelial cells in vitro . These factors were analyzed subsequently to determine whether they predicted mortality in a mouse model of hematogenously disseminated candidiasis . By proportional hazard analysis, the relative risk of death was 5.6-fold higher (95% confidence interval, 1.672 to 18.84 {P < 0.005}) in the mice infected with the higher-phospholipase-secreting strains than in the low-phospholipase secretors . None of the other putative virulence factors predicted mortality . Characterization of phospholipases secreted by three of the blood isolates showed that these strains secreted both phospholipase B and lysophospholipase-transacylase activities . These results implicate extracellular phospholipase as a virulence factor in the pathogenesis of hematogenous infections caused by C . albicans.

Infect Immun, 1995 May, 63(5), 1887 - 92
Expression of Candida albicans SAP1 and SAP2 in experimental vaginitis; De Bernardis F et al.; Several strains of Candida albicans were compared for their ability to cause vaginal infection in a rat model, and their vaginopathic potentials were correlated with the expression of two aspartyl proteinases genes (SAP1 and SAP2) and adherence in vivo to the vaginal epithelium . Dot blot reactions and Northern blot analysis with RNA extracted from the vaginal fluid of rats infected with the highly vaginopathic strains H12 and 10261 demonstrated the expression of both SAP1 and SAP2 during the first week of infection . In contrast, neither gene was expressed during infection by a nonvaginopathic strain (N), even though the organism could be recovered during the first 24 h postinfection . A moderately vaginopathic strain (P) also expressed both genes, but the level of SAP1 mRNA appeared to decrease prior to that of SAP2 . Neither gene was expressed, even by the highly vaginopathic strains, after the first week of infection, concomitant with a decrease in the number of organisms recovered from the vaginas . Analysis of in vivo adherence showed that the nonvaginopathic strain (N) adhered to vaginal epithelial cells less readily than the highly vaginopathic strain (H12) and moderately vaginopathic strain (P) . Thus, in addition to its inability to express SAP1 and SAP2 in vivo, the nonvaginopathic strain does not colonize host cells to the same extent as the other strains tested . Our results demonstrate the early in vivo expression of two aspartyl proteinase gene during candidal vaginitis and suggest its association with the establishment of a vaginal infection.

Infect Immun, 1995 May, 63(5), 1806 - 9
Differential susceptibility of yeast and hyphal forms of Candida albicans to macrophage-derived nitrogen-containing compounds; Blasi E et al.; Candida albicans is a dimorphic fungus capable of transition from the yeast form (Y-Candida) to the hyphal form (H-Candida) . Both Y-Candida and H-Candida are known to be growth inhibited by murine macrophages (M phi) in vitro . In the present report, we demonstrate that M phi exposed to interferon gamma (IFN-gamma) plus lipopolysaccharide (LPS) show enhanced anti-Y-Candida and anti-H-Candida activities . To further investigate the phenomenon, Y-Candida and H-Candida were assessed for susceptibilities to M phi-derived supernatants . Only the growth of H-Candida, and not that of Y-Candida, is impaired by cell-free supernatants from M phi treated with IFN-gamma plus LPS . In contrast, no H-Candida growth inhibition occurs when supernatants from M phi exposed to IFN-gamma plus LPS in the presence of NG-monomethyl-L-arginine, an inhibitor of nitric oxide (NO) synthesis, are employed . Finally, supernatants from M phi incubated with sodium nitroprusside, an NO-generating agent, also show anti-H-Candida activity . In conclusion, these results indicate that H-Candida but not Y-Candida is susceptible to extracellular antifungal mechanisms employed by M phi, which likely involve stable nitrogen-containing compounds.

Pediatrics, 1995 May, 95(5), 682 - 7
Invasive fungal dermatitis in the < or = 1000-gram neonate; Rowen JL et al.; OBJECTIVE . In 1991, we noted the emergence amongst our extremely low birth weight neonates of a new clinical entity, invasive fungal dermatitis, characterized by erosive, crusting lesions and a high rate of subsequent systemic fungal infection . We sought to define this condition and examine potential risk factors . METHODS . Sixteen neonates with invasive fungal dermatitis were seen during a 2-year period in three Baylor College of Medicine affiliated intensive care nurseries . Seven were confirmed cases, with skin biopsy evidence of invasion beyond the stratum corneum . Nine had a consistent clinical course and a positive potassium hydroxide examination of skin scrapings or isolation of fungi from skin or systemic cultures . Three controls were matched to each case by hospital, date of admission, and birth weight . Data was collected by retrospective chart review . RESULTS . Invasive fungal dermatitis occurred in 5.9% of at-risk infants . Case patients had a mean birth weight of 635 g and developed skin lesions at a mean age of 9 days (range, 6 to 14) . Candida albicans was the most commonly implicated pathogen, but other Candida species, Aspergillus, Trichosporon beigelii, and Curvularia were also seen . Disseminated infection occurred in 69%, all due to Candida sp . Case patients were significantly more premature than controls (mean gestation, 24.4 vs 25.9 weeks) and were more likely to be delivered vaginally (81% vs 50%) . Postnatal steroids were administered to cases (81%) more often than controls (46%) . Case patients had more prolonged hyperglycemia (as assessed by insulin administration) than controls (mean 4.3 vs 2.0 days) . CONCLUSIONS . Invasive fungal dermatitis is a disease of the smallest, most immature neonates and is associated with vaginal birth, steroid administration, and hyperglycemia . We speculate that the skin serves as a portal of entry for colonizing fungal species and may thus lead to disseminated infection . Methods to improve skin barrier function may be useful in preventing this disorder.

J Med Vet Mycol, 1995 May-Jun, 33(3), 205 - 7
Molecular identification of Candida albicans; Weissman Z et al.; The benomyl resistant gene (BenR) found in Candida albicans, but not in other species of Candida, was used as a probe for the identification of C . albicans in clinical specimens . The utility of this probe to detect this species was demonstrated by Southern and dot-blot analysis, and by PCR . The possible use of this gene in C . albicans typing by the RFLP method is also demonstrated.

J Med Vet Mycol, 1995 May-Jun, 33(3), 201 - 3
Highly sensitive detection of fungal antigens by ultrasound-enhanced latex agglutination; Grundy MA et al.; Treatment with ultrasound has been employed to greatly enhance the sensitivity of commercially available latex agglutination tests for fungal antigens . This 5 min procedure detects 40 pg ml-1 of Candida albicans mannan and 70 pg ml-1 of Aspergillus fumigatus galactomannan, a 250 and 500-fold improvement respectively over conventional agglutination test sensitivities . The ultrasound-enhanced test offers the possibility of improved diagnosis and management of patients with systemical candidosis or invasive aspergillosis.

J Med Vet Mycol, 1995 May-Jun, 33(3), 191 - 5
Combined effect of amphotericin B and flucytosine on hyphal growth of Candida albicans estimated at a single hypha level; Oh KB et al.; The in vitro efficacy of amphotericin B and flucytosine, separately and in combination, against the hyphal growth of Candida albicans was evaluated in situ using an automatic analysing system . A colony of C . albicans was in contact with a glucose-salt medium supplemented with biotin plus calf serum (GS medium) and GS medium containing the antifungal agent, in sequence . Minimum inhibitory concentrations at single hyphal level (S-MIC) were determined based on the response that was measured . Amphotericin B S-MICs ranged from 0.8 to 0.4 microgram ml-1, and S-MIC values > 102.4 micrograms ml-1 were obtained with flucytosine when the two agents were used independently . When the two agents were used in combination, however, a synergistic interaction between the two agents at concentrations below their individual S-MICs was observed.

J Interferon Cytokine Res, 1995 May, 15(5), 421 - 9
Defective expression of interferon-gamma, granulocyte-macrophage colony-stimulating factor, tumor necrosis factor alpha, and interleukin-6 in activated peripheral blood lymphocytes from glioma patients; Urbani F et al.; The ability of a mannoprotein antigen from Candida albicans (MP) or interleukin-2 (IL-2) to induce cytokines in cultures of peripheral blood mononuclear cells (PBMC) of glioma patients and healthy controls was evaluated by mRNA expression and by protein secretion . The subjects studied were all responsive to both MP and IL-2, as assayed by lymphoproliferation of PBMC cultures . In control subjects, MP and IL-2 were strong inducers of IFN-gamma, IL-1 beta, TNF-alpha, and GM-CSF mRNA expression, but only MP was able to induce considerable levels of IL-6 and IL-2 mRNA expression . In MP-activated PBMC from glioma subjects, a highly defective IFN-gamma, together with a significant reduction in TNF-alpha and GM-CSF mRNA expression, was observed . This impairment was paralleled by a decreased accumulation of IL-6 and IL-2 mRNA . The pattern of cytokine mRNAs in IL-2-activated PBMC of glioma patients confirmed the impairment of IFN-gamma mRNA expression paralleled by a reduction in IL-6, TNF-alpha and GM-CSF mRNA, compared with healthy subjects . Coherently, in PBMC cultures from glioma patients, there was a clear-cut decrease in the secretion of IL-6 and TNF-alpha and especially of IFN-gamma compared with healthy controls . No or very low levels of IL-4, IL-10, and TGF-beta 2 mRNA expression were detected in PBMC cultures of both glioma and control populations, irrespective of the activation conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

J R Soc Med, 1995 May, 88(5), 258 - 63
Fungal feeding-line infections: beware the eyes and teeth; Nightingale JM et al.; Twenty-four fungal feeding-line infections occurred in 17 patients during 1984-1992 . Thirteen were receiving long-term home parenteral feeding and, in them, the first infection occurred after a median of 30 months (range 1-120) continuous feeding with a line that had been in situ for a median of 20 months (range 1-37) . Four were receiving short-term feeding through a line that had been inserted 1-2 months previously . At the time of the first infection all patients were febrile and most were anaemic (15/16), however a leucocytosis was rare (three of 16) . The fungi isolated were Candida albicans(6), Candida parapsilosis(5), Candida glabrata(2), Candida guillermondii(2) and other species (2) . In 16 patients, the feeding-line was removed at the time of the first infection and no other treatment was given, and no other complications occurred in eight (50%) of these . In 11, the line was reinserted a median of 7 days after removal (range 1-11) . Four patients (24%) developed a Candida infection of the eye 1-8 weeks after the diagnosis, uveitis (2) and endophthalmitis (2) which, in one patient, led to complete blindness in one eye . Two patients had recurrent infections which began within a month of dental therapy . In one, the infections stopped after dental extractions and, in the other, after a dental clearance . An ophthalmoscopic examination should be performed in all patients with a fungal feeding-line infection . Recurrent candidal infections may have a dental origin.

Immunology, 1995 May, 85(1), 153 - 9
Mannose binding protein is involved in first-line host defence: evidence from transgenic mice; Tabona P et al.; Mannose binding protein (MBP) is a calcium-dependent C-type lectin secreted by the liver which seems to be an important component of innate or natural immunity . We have investigated the effects of Candida albicans and thioglycolate injection into transgenic mice bearing the human MBP gene . The transgenes contained a 15 kb fragment of the MBP gene which included the complete coding sequence . Northern blot hybridization showed human MBP mRNA transcripts in the liver of two transgenic lines with low and high copy number respectively . Western blot analysis showed the presence in serum of human MBP which associated into the higher multimeric forms which are capable of activating complement . Enzyme-linked immunosorbent assays (ELISA) showed that serum human MBP concentrations in the transgenes (1.90 +/- 0.16 mg/l, mean +/- SEM) were about twice as high as the levels in man . The serum concentration of MBP A, which is the mouse homologue of MBP, (13.9 +/- 0.45 mg/l) was about seven times that of human MBP . Intravenous injection of Candida albicans caused the serum human MBP level to fall by more than 50% in the first hour and then slowly recover, but it did not return the initial value by 72 hr . Candida injection caused a 25% fall in serum mouse MBP A in the first hour which then rose to supranormal levels by 72 hr . Following Candida injection mouse MBP A mRNA concentrations increased over 72 hr in contrast to human MBP mRNA which remained constant in both transgenic lines . These data indicate that the human MBP gene fragment in the transgene did not include the regulatory elements of the gene . Total haemolytic complement activity and C3 concentrations also fell immediately after Candida and thioglycolate injection while the concentrations of mannose specific immunoglobulin G (IgG) and immunoglobulin M (IgM) did not fall . The data indicate that mannose binding protein plays an important role in the initial stages of defence against infection which, in this model, is quantitatively greater than that of mannose-specific IgG and IgM antibodies . Mannose binding protein is probably most important in defense of previously unexposed and non-immune hosts.

J Clin Microbiol, 1995 May, 33(5), 1283 - 8
Isolation and characterization of species-specific DNA probes from Taenia solium and Taenia saginata and their use in an egg detection assay; Chapman A et al.; Cysticercosis results from ingestion of the eggs of the tapeworm Taenia solium . Reduction of the incidence of human and swine cysticercosis requires identification and treatment of individuals who carry the adult tapeworm . T . solium and Taenia saginata eggs cannot be differentiated on the basis of morphology; thus, in order to improve existing methods for the diagnosis of taeniasis, we have developed highly sensitive, species-specific DNA probes which differentiate T . solium and T . saginata . Recombinant clones containing repetitive DNA sequences which hybridize specifically with genomic DNAs from either species were isolated and characterized . T . solium-specific DNA sequences contained complete and truncated forms of a tandemly repeated 158-bp DNA sequence . An unrelated T . saginata DNA sequence was also characterized and shown to encode a portion of the mitochondrial cytochrome c oxidase I gene . T . solium- and T . saginata-specific DNA probes did not hybridize in dot blot assays either with genomic DNA from the platyhelminths Taenia hydatigena, Taenia pisiformis, Taenia taeniaeformis, Echinococcus granulosus, and Schistosoma mansoni or with genomic DNA from other eukaryotes, including Saccharomyces cerevisiae, Candida albicans, Cryptosporidium parvum, Entamoeba histolytica, Trypanosoma gambiense, Trypanosoma brucei, and Giardia lamblia, Caenorhabditis elegans, and human DNA . By using these T . solium and T . saginata DNA probes, a rapid, highly sensitive and specific dot blot assay for the detection of T . solium eggs was developed.

J Clin Microbiol, 1995 May, 33(5), 1238 - 42
Molecular typing of Candida albicans in oral candidiasis: karyotype epidemiology with human immunodeficiency virus-seropositive patients in comparison with that with healthy carriers; Lupetti A et al.; Candida albicans organisms isolated from the oral cavities of healthy carriers (26 individuals) and compromised hosts (40 human immunodeficiency virus {HIV}-seropositive patients, all showing symptomatic oral candidiasis) were compared by resolving chromosome-sized DNA molecules into electrophoretic karyotypes . Seven- to 10-band electrophoretic patterns were obtained, with significant and reproducible differences in the distributions of the DNA bands . Seven distinct classes were identified and were designated type a (8 bands), type b (8 bands), type c (7 bands), type d (9 bands), type x (10 bands), type y (10 bands), and type z (9 bands) . Four of these (types a to d) were the most representative within all of the isolated strains (95.5%), and the other three (types x to z) were observed only once in three HIV-seropositive individuals (4.5%) . Only types b and c were isolated from healthy carriers, with the percentage of their isolation being 61.5 and 38.5%, respectively, while all the described karyotypes were isolated from HIV-seropositive patients, with type b being the most frequent (45%); this was followed by types c (25%), a (15%), and d (7.5%) . The prevalence of type b and c karyotypes in HIV-infected individuals, as well as in healthy carriers, suggests that commensal strains in the oral cavities of healthy individuals may become the prevalent agents of subsequent oral candidiasis in compromised hosts . However, replacement of the original, commensal strain, if there is one, cannot be excluded in a compromised host, although strain replacement may be more reasonably hypothesized for types a and d, since only these types were isolated at a relative high percentage from the oral lesions of HIV-infected individuals.

J Clin Microbiol, 1995 May, 33(5), 1223 - 30
Evidence for nosocomial transmission of Candida albicans obtained by Ca3 fingerprinting; Schmid J et al.; The moderately repetitive sequence Ca3 was used to fingerprint Candida albicans isolates from 32 patients hospitalized for more than 3 days, 17 recent admissions or outpatients, and 8 recently readmitted patients and 10 commensal isolates from the community in Wellington, New Zealand, plus isolates from 21 hospitalized patients, 26 outpatients or recent admissions, 4 recently readmitted patients, and 10 healthy individuals in the community in Auckland, New Zealand . In Wellington, isolates from patients hospitalized in Wellington Hospital for more than 3 days were genetically significantly less diverse than were isolates from outpatients or recent admissions or isolates from healthy individuals in the community . In addition, two clusters of genetically similar strains were isolated from hospitalized patients significantly more often than from other individuals . These observations provide evidence (albeit indirectly) for nosocomial transmission of hospital-specific C . albicans strains . In contrast, no indication of hospital-specific transmission of C . albicans was found in Auckland Hospital . Since these results were obtained under conditions in which no candidiasis outbreak occurred in either hospital, they also suggest that Ca3 fingerprinting may be a useful tool in preventive nosocomial infection control programs, allowing assessment of the extent of C . albicans transmission occurring in a hospital.

J Clin Microbiol, 1995 May, 33(5), 1129 - 35
Cluster of oral atypical Candida albicans isolates in a group of human immunodeficiency virus-positive drug users; Boerlin P et al.; Twenty-one chlamydospore-forming and germ tube-positive Candida albicans clinical isolates from 15 human immunodeficiency virus (HIV)-positive and 3 HIV-negative patients were examined by two different genetic methods . Multilocus enzyme electrophoresis and hybridization with the C . albicans-specific Ca3 probe showed that such isolates can be split into two genetically distinct groups that can be clearly distinguished . One group mainly contained strains with atypical sugar assimilation patterns and could be distinguished from the other group by the absence of intracellular beta-glucosidase activity . All 13 strains belonging to this group were isolated from the oral cavities of asymptomatic HIV-positive drug users and may be less pathogenic than the eight strains from the other group isolated either from HIV-positive patients with oropharyngeal candidiasis or from HIV-negative patients with invasive candidiasis.

Arzneimittelforschung, 1995 May, 45(5), 620 - 3
Synthesis and antifungal activities of some new triazolylacetophenone derivatives; Ertan R et al.; In this study some alpha-(1,2,4-triazolyl)acetophenone derivatives were synthesized by the condensation of various triazole and alpha-bromoacetophenone rings . The structures of the compounds have been elucidated by UV, IR, 1H-NMR, mass spectra and elementary analysis . The in vitro antifungal activity of the compounds were investigated against some yeast-like fungi (Candida albicans, C . parapsilosis, C . stellatoidea and C . pseudotropicalis) and moulds such as Trichophyton rubrum and T . mentagrophytes by the tube dilution method and MIC (minimal inhibitory concentration), MFC (minimal fungicidal concentration) values were determined . Compound A16 was significantly more effective than the other compounds . The results obtained for antifungal activity against moulds were not significant.

Kansenshogaku Zasshi, 1995 May, 69(5), 590 - 6
{Effect of erythromycin on macrophage functions}; Xu G et al.; Recently, it has been suggested that macrolide antibiotics act as immunomodulators . In this study, we evaluated the effect of EM on macrophage function . We used the mouse macrophage cell line, J774.1 . The following direct effects of EM on macrophage function were evaluated: chemotaxis to EM, chemokinetic effect by EM, and the effect of EM on macrophage growth . In order to examine the indirect effects of EM on macrophage functions, we preincubated macrophages with several concentrations of EM and then removed the EM . Thereafter, the phagocytosis of beads, cytocidal activity against Candida albicans, and chemotaxis to lipopolysaccharide were evaluated . EM (at the concentration between 0.04 and 0.2 microgram/ml) directly stimulated macrophage chemotaxis and chemokinesis . In addition, EM dose-dependently stimulated the growth of macrophages . EM pretreatment (for 4 hours at the contractions between 0.04 and 0.2 microgram/ml) stimulated macrophage phagocytosis, cytocidal activity against Candida albicans, and chemotaxis to lipopolysaccharide . These results suggest that EM has direct and indirect effects on macrophage functions.

Kansenshogaku Zasshi, 1995 May, 69(5), 582 - 9
{Protective effect of human macrophage colony-stimulating factor on fungal infection (2) . In vitro effect of human macrophage colony-stimulating factor on systemic aspergillosis and in vitro effect on the activities of macrophage}; Fujita H et al.; We studied the protective effect of human macrophage colony-stimulating factor (M-CSF) of fungal infection due to systemic aspergillosis in normal mice . We also examined the effect of M-CSF against the activities of mouse peritoneal macrophage which were relating to the phagocytosis, the killing, the production of superoxide after contacting with phorbol myristate acetate and the production of nitric oxide after contacting with interferon-gamma in vitro . M-CSF improved the median survival time and the survival rate of systemic aspergillosis . Combination therapy with M-CSF and amphotericin-B (AMPH-B) showed the therapy with either M-CSF or AMPH-B alone . M-CSF enhanced the activities of phagocytosis and the killing of ingested Candida albicans H and spores of Aspergillus fumigatus K by macrophage . Furthermore, M-CSF promoted the production of superoxide and nitric oxide in macrophage . These results indicate that M-CSF can enhance the fungicidal activity of macrophages by activation in vivo, thereby preventing the dissemination of fungal infection.

Eur J Clin Microbiol Infect Dis, 1995 May, 14(5), 406 - 11
Differential and enrichment media for selective culture and recognition of yeast species from clinical material; Louwagie B et al.; An enrichment medium was developed and evaluated for isolation of fluconazole-resistant minority yeast species in a hospital setting . The enrichment medium was made by adding fluconazole (10 micrograms/ml) to yeast nitrogen base/glucose broth . Under laboratory conditions the broth permitted detection of 20 of 20 Candida krusei isolates and 20 of 20 Candida glabrata isolates in mixtures with Candida albicans even when the Candida albicans cells outnumbered those of the other species by 1000:1 . Results of culture on the enrichment medium were compared with those obtained on routine agar media and on a yeast differential agar which facilitates detection of mixed yeast species by their colony colours . Only one Candida glabrata isolate was detected on the enrichment broth but not found on routine culture of 68 yeast-positive clinical specimens . However, bacterial over-growth in some broths may have retarded the appearance of other yeast isolates . On the yeast differential agar, 20 clinical specimens were found to contain mixtures of yeast species compared with only 2 on routine culture.

Int Immunol, 1995 May, 7(5), 785 - 96
Immunoprotection against systemic candidiasis in mice; Tavares D et al.; We have previously described an immunosuppressive B cell mitogenic (ISM) protein, p43, produced by Candida albicans, which plays an important role in the survival of the microorganism in the host . The N-terminal amino acid sequence of p43 was found to be different from all amino acid sequences registered in updated protein databanks . Immunization of BALB/c mice with p43 partially neutralized the biological effects of this protein, namely depletion of bone marrow pre-B and B cells, the increased numbers of total and large B and CD4+ lymphocytes, and the non-specific polyclonal response of splenic IgG2a-, IgG2b- and IgM-secreting plaque forming cells . Immunization of BALB/c mice with p43 fully protected the mice against the fungal infection . In contrast, immunization with C . albicans sonicates (Cs) was not protective . Our data indicated that specific antibodies against p43 protected, whereas those against Cs facilitated C . albicans infection . Thus, the ratio between anti-p43 and anti-Cs antibody titres was much lower in the non-protected mice (Cs-immunized and control non-immunized) than in p43-immunized mice . Moreover, passive administration of specific anti-p43 antibodies significantly protected against fungal infection, whereas passive administration of specific anti-Cs antibodies markedly increased the susceptibility to C . albicans infection . These observations are discussed on the basis of alternative approaches of immunointervention.

Biochem Mol Biol Int, 1995 May, 35(6), 1215 - 22
Status of membrane lipids and amino acid transport in morphological mutants of Candida albicans; Koul A et al.; The phospholipid composition of various morphological mutants of Candida albicans revealed a complete absence of phosphatidylinositol (PI) from plasma membranes of those cells which completely lacked mycelial growth . No other phospholipid was found to be specific to morphogenesis . The plasma membrane fractions isolated from mutants were more rigid than its wild type as was evident from their unsaturation index and fluorescence polarization measurements . The enhanced membrane rigidity of mutant cells was noted regardless whether the cells could grow only as mycelia or in their budding forms . Although some amino acids are considered to affect the morphological transition of C . albicans, this was not reflected in the transport activities of L-proline, L-alanine, L-lysine and L-glutamic acid.

Pharm Res, 1995 May, 12(5), 649 - 52
Primary interactions of three quaternary ammonium compounds with blastospores of Candida albicans (MEN strain); Schep LJ et al.; The absorption of three quaternary ammonium compounds (QAC), cetylpyridinium chloride, cetrimide and benzalkonium chloride, onto the surface of blastospores of Candida albicans (MEN strain) was examined at room temperature . Equilibrium uptake occurred in less than 30 seconds for cetylpyridinium chloride and cetrimide whereas 5 min contact time was required for benzalkonium chloride . The adsorption of all three agents may be mathematically described as Langmuirian and hence a concentration-dependent formation of drug-monolayer on the surface of the blastospore occurred . From this the number of molecules adsorbed onto the surface of a single blastospore was calculated to be 1.33 x 10(12), 3.17 x 10(12) and 2.32 x 10(12) for cetylpyridinium chloride, cetrimide and benzalkonium chloride, respectively . These dissimilarities are most likely due to differences in the orientations of both the cationic nitrogen atom and the accompanying lipophilic portions of each QAC at the blastospore surface . Relating these observations to the known antiadherence effects of cetylpyridinium chloride and cetrimide, it can be concluded that monolayer coverage of the blastospore surface with QAC does not account for the observed reduced adherence . This suggests that the anti-adherence effects are due to either direct interaction with, or steric blockade of, adhesions on the blastospore surface.

Yeast, 1995 Apr 15, 11(4), 301 - 10
Cloning and heterologous expression of the Candida albicans gene PMI 1 encoding phosphomannose isomerase; Smith DJ et al.; Using a DNA fragment derived from the Saccharomyces cerevisiae phosphomannose isomerase (PMI) structural gene as a probe against a random ordered array library of genomic DNA from the pathogenic fungus Candida albicans, we have cloned the C . albicans PMI 1 gene . This gene, which is unique in the C . albicans genome, can functionally complement PMI-deficient mutants of both S . cerevisiae and Escherichia coli . The DNA sequence of the PMI 1 gene predicts a protein with 64.1% identity to PMI from S . cerevisiae . Sequential gene disruption of PMI 1 produces a strain with an auxotrophic requirement for D-mannose . The heterologous expression of the PMI 1 gene at levels up to 45% of total cell protein in E . coli leads to partitioning of the enzyme between the soluble and particulate fractions . The protein produced in the soluble fraction is indistinguishable in kinetic properties from the material isolated from C . albicans cells.

FEMS Microbiol Lett, 1995 Apr 15, 128(1), 95 - 100
Cloning of a DNA fragment encoding part of a 70-kDa heat shock protein of Candida albicans; Eroles P et al.; Immunoscreening of a mycelial expression library with polyclonal antibodies raised against mycelial cell wall resulted in the detection of a cDNA encoding a heat shock protein of Candida albicans . Sequence analysis of a 0.8-kb cDNA subclone, 2M-1, revealed an open reading frame encoding 244 amino acids . Southern blot analysis with this fragment as a probe demonstrated hybridization to C . albicans DNA . Northern analysis showed a substantial increase in 2M RNA expression levels after cells were subjected to heat shock . Western blot analysis with 2M monospecific antibodies recognized a 70-kDa protein which was present in membrane particles and cytosolic fractions.

Cell Immunol, 1995 Apr 15, 162(1), 105 - 13
Suppression of the functional activity of IL-2-activated lymphocytes by CGRP; Wang X et al.; CGRP is a 37-amino acid neuropeptide found in the central and peripheral nervous systems as well as the nerve endings in lymphoid organs . Specific CGRP receptors are present on both T and B lymphocytes . There is increasing evidence that CGRP plays a role in regulation of the immune response . However, few investigations have examined the effects of CGRP on lymphocyte effector functions . In this report, CGRP (0.1 nM-1 microM) was shown to cause concentration-dependent inhibition of IL-2-activated lymphocyte growth inhibition of the fungus Candida albicans and cytotoxic activity for tumor cells . Maximum inhibition of lymphocyte activity by CGRP was 47.4% for the hyphae of C . albicans, 44.8% for a natural killer cell susceptible cell line, and 52.9% for a natural killer cell-resistant cell line . CGRP-mediated inhibition of lymphocyte function was mimicked by 8-bromo-cAMP (1 mM) and was correlated in a concentration-dependent manner with an increase in intracellular levels of cAMP . These increases were potentiated by pretreatment of the lymphocytes with 3-isobutyl-1-methylxanthine (0.5 mM, 10 min), an inhibitor of the cAMP phosphodiesterase . hCGRP 8-37, a selective blocker of the CGRP1 receptor, abrogated the effect of CGRP on lymphocyte function and on intracellular cAMP level elevation induced by rCGRP . CGRP had no direct effect on the capacity of IL-2-activated lymphocytes to adhere to the hyphae of C . albicans . However, both CGRP and 8-bromo-cAMP diminished the capacity of the lymphocytes to release cytoplasmic granular content when stimulated by the hyphae of C . albicans . These data show that CGRP inhibits functional activity of IL-2-activated lymphocytes and suggest that hCGRP8-37 may be a useful tool for assessing the role of CGRP in the immune system.

Gene, 1995 Apr 14, 156(1), 133 - 8
Analysis and expression of the Candida albicans FAS2 gene; Southard SB et al.; Candida albicans (Ca) FAS2, the gene encoding the alpha-subunit of fatty acid synthase (FAS), has been isolated and characterized . Saccharomyces cerevisiae (Sc) FAS2 was used as a probe to screen genomic libraries of Ca, strain 4918 . Clones were obtained that contained all but the first 1 kb of the gene . The 5' end, as well as upstream sequences, were subsequently isolated by PCR . The Ca FAS2 gene is comprised of an open reading frame (ORF) of 5655 bp that is free of introns and encodes a 207, 587-Da protein of 1885 amino acids . The Ca FAS2 gene and the encoded polypeptide exhibit 70 and 67% homology with the Sc FAS2 gene and protein, respectively . The gene specifies a single transcript of approx . 6 kb, and transcript levels are regulated independently of morphogenesis . Chromosome analysis localized the gene to chromosome 3 . In addition, no differences were noted when comparing the FAS2 sequence, that encompasses the cerulenin-binding domain of FAS, between strain 4918 and two derived cerulenin-resistant (CerR) mutants . Thus, alteration within this region is not responsible for the increased CerR of the mutant strains.

Rev Argent Microbiol, 1995 Apr-Jun, 27(2), 81 - 9
{In vitro sensitivity tests for yeasts: evaluation of a micromethod}; Rodero LL et al.; A microdilution antifungal susceptibility test for yeasts was evaluated, based on the macro broth dilution method standardized by the National Committee for Clinical Laboratory Standards (NCCLS) Subcommittee on Antifungal Susceptibility Testing . Both methods were compared using six reference strains with different patterns of susceptibility to the following antifungal drugs: amphotericin B (AMB), flucytosine (5FC), fluconazole (FCZ), itraconazole (ITZ), ketoconazole (KTZ) and miconazole (MCZ) . Minimal inhibitory concentration (MIC) results obtained by both methods differed only in 1 or 2 dilutions . Microdilution MIC's determined as visual endpoints and as quantitative measurement of 80% inhibition of relative growth showed a significant correlation (p < 0.001) for AMB, 5FC, FCZ, MCZ and ITZ, conversely no correlation (p = 1.00) for KTZ was observed as determined with 47 local isolates of Candida albicans.

Exp Mol Pathol, 1995 Apr, 62(2), 109 - 17
Rapid destruction of skeletal muscle fibers by mycelial growth forms of Candida albicans; Ashman RB et al.; The process of tissue invasion by the yeast Candida albicans after subcutaneous inoculation into the footpad of mice has been examined by light and electron microscopy . The blastospores germinated within 2 hr after injection . Hyphal and, to a lesser extent, pseudohyphal elements were observed penetrating into the adjacent striated muscle fibers and the endothelial cells of capillaries and destroying the collagen bundles of the interstitium . Electron microscopy showed evidence of structural tissue degradation associated with the mycelial elements, particularly at the apical tip . This was presumably due to fungal enzymatic activity . By 3 hr after infection, polymorphonuclear leucocytes had engulfed both the extracellular and intracellular mycelia, and often there was structural evidence of mycelial degradation . The results provide morphological evidence that initial tissue damage following subcutaneous infection is caused by the mycelial growth form.

Braz J Med Biol Res, 1995 Apr, 28(4), 477 - 83
Increased clearance of Candida albicans from the peritoneal cavity of mice pretreated with concanavalin A or jacalin; Felipe I et al.; We have studied the role of polymorphonuclear leukocytes and macrophages in the clearance of Candida albicans from the peritoneal cavity of Swiss mice after treatment with jacalin or concanavalin A (Con-A) . Mice (25-30 g, N = 7 per group) received jacalin or Con-A (500 micrograms/0.5 ml PBS) intraperitoneally 96 h before intraperitoneal inoculation of 5 x 10(7) yeast cells . The clearance of Candida from the peritoneal cavity was complete 24 h after inoculation for animals pretreated with jacalin and 48 h after inoculation for animals pretreated with Con-A, whereas a reduction to 4 x 10(4) yeast cells/cavity occurred in control animals 48 h after inoculation . Pretreatment with jacalin or Con-A reduced the recovery of C . albicans from spleen, kidney and liver 10- to 80-fold compared to control animals . Pretreatment with the lectins increased the number of phagocytic cells in the peritoneal exudate 5- to 10-fold and their candidacidal activity was increased 6-fold compared to controls . These data explain the increased rate of clearance and reduced yeast dissemination to the viscera of lectin treated mice.

J Bacteriol, 1995 Apr, 177(7), 1772 - 9
Genetic organization and mRNA expression of enolase genes of Candida albicans; Postlethwait P et al.; In previous work, we cloned a Candida albicans cDNA for the glycolytic enzyme enolase and found a single, abundant enolase transcript on Northern (RNA) blots and a single protein on immunoblots, using antiserum raised against a recombinant enolase fusion protein . Because C . albicans enolase is abundantly produced during infection and elicits strong host immune responses, the mechanisms regulating enolase production are important for understanding the growth of C . albicans in vivo . To obtain more information on enolase gene expression by C . albicans, we used the enolase cDNA clone to investigate the genetic organization of enolase genes and the steady-state levels of enolase mRNA under several growth conditions . Gene disruption techniques in combination with Southern blot analyses of genomic DNA showed the presence of two enolase gene loci that could be distinguished by the locations of ClaI and Mn/I sites in their 3' flanking regions . Enolase steady-state mRNA levels were greatest during the middle phase of the logarithmic growth curve and were low during stationary phase . Minimal differences in enolase mRNA levels between yeast cells and hyphae were found . Propagation of C . albicans in glucose did not cause increased enolase mRNA levels compared with growth in a nonfermentable carbon source (pyruvate) . It was concluded that two gene loci exist for C . albicans enolase and that enolase mRNA is constitutively produced at high levels during active metabolism.

Mol Cell Biol, 1995 Apr, 15(4), 2197 - 206
Functional characterization of the MKC1 gene of Candida albicans, which encodes a mitogen-activated protein kinase homolog related to cell integrity; Navarro-Garcia F et al.; Mitogen-activated protein (MAP) kinases represent a group of serine/threonine protein kinases playing a central role in signal transduction processes in eukaryotic cells . Using a strategy based on the complementation of the thermosensitive autolytic phenotype of slt2 null mutants, we have isolated a Candida albicans homolog of Saccharomyces cerevisiae MAP kinase gene SLT2 (MPK1), which is involved in the recently outlined PKC1-controlled signalling pathway . The isolated gene, named MKC1 (MAP kinase from C . albicans), coded for a putative protein, Mkc1p, of 58,320 Da that displayed all the characteristic domains of MAP kinases and was 55% identical to S . cerevisiae Slt2p (Mpk1p) . The MKC1 gene was deleted in a diploid Candida strain, and heterozygous and homozygous strains, in both Ura+ and Ura- backgrounds, were obtained to facilitate the analysis of the function of the gene . Deletion of the two alleles of the MKC1 gene gave rise to viable cells that grew at 28 and 37 degrees C but, nevertheless, displayed a variety of phenotypic traits under more stringent conditions . These included a low growth yield and a loss of viability in cultures grown at 42 degrees C, a high sensitivity to thermal shocks at 55 degrees C, an enhanced susceptibility to caffeine that was osmotically remediable, and the formation of a weak cell wall with a very low resistance to complex lytic enzyme preparations . The analysis of the functions downstream of the MKC1 gene should contribute to understanding of the connection of growth and morphogenesis in pathogenic fungi.

Infect Immun, 1995 Apr, 63(4), 1373 - 9
Expression of surface hydrophobic proteins by Candida albicans in vivo; Glee PM et al.; Candida albicans modulates cell surface hydrophobicity during growth and morphogenesis in vitro . To determine if surface hydrophobicity is expressed during pathogenesis, we generated a polyclonal antiserum against yeast hydrophobic proteins . The antiserum was then used for indirect immunofluorescence analysis of tissues from mice colonized and chronically infected with C . albicans . Results demonstrated that yeast hydrophobic proteins are exposed on fungal cells present in host tissues . The polyclonal antiserum distinguished between hydrophobic and hydrophilic cell surfaces in vitro and gave similar staining patterns and intensities for C . albicans cells in vivo . Of the yeast forms present within tissue lesions, approximately half exhibited moderate to intense immunofluorescence with the antiserum . Immunoblot analysis indicated that antigens recognized by the antiserum are predominantly low-molecular-mass hydrophobic proteins that are expressed by different C . albicans isolates and are expressed regardless of growth temperature . Taken together, the immunofluorescence and immunoblot analyses of antigens indicate that C . albicans displays surface hydrophobic proteins during pathogenesis and these proteins are available for hydrophobic interactions with host tissues . The effect of hydrophobic protein exposure on the virulence of C . albicans is discussed.

Infect Immun, 1995 Apr, 63(4), 1253 - 7
Differential susceptibility of yeast and hyphal forms of Candida albicans to proteolytic activity of macrophages; Blasi E et al.; The dimorphic transition of Candida albicans from the yeast (Y-Candida) to the hyphal (H-Candida) form is a complex event whose relevance in fungal pathogenicity is still poorly understood . Using a cloned macrophage (M phi) cell line (ANA-1), we have previously shown that a M phi can discriminate between the two fungal forms, eliciting different secretory responses . In the present study, we investigated the susceptibility of Y-Candida and H-Candida to M phi proteolytic activity . In particular, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot (immunoblot) techniques were employed to analyze the patterns of lyticase proteinaceous extracts from cell walls of Y-Candida and H-Candida which had been unexposed or exposed to ANA-1 M phis for 3 h . Silver staining allowed detection of a complex protein pattern in both forms of C . albicans, qualitatively and quantitatively differing from each other, mainly at molecular masses below 106 kDa . Western blot staining with anti-C . albicans mannan antibodies and convalescent-phase sera of mice previously infected systemically or intracerebrally with C . albicans showed that, after contact with M phis, Y-Candida but not H-Candida proteinaceous cell wall components are profoundly modified, with substantial reduction and/or disappearance of many bands . Our experimental approach provides initial insights into the differential susceptibility of Y-Candida and H-Candida to the proteolytic activity of M phis.

Immunol Cell Biol, 1995 Apr, 73(2), 125 - 33
Humoral immune responses to systemic Candida albicans infection in inbred mouse strains; Costantino PJ et al.; The protective role of humoral antibodies in the resolution of systemic candidiasis remains controversial . Investigation of the humoral immune responses in mouse strains of varying susceptibility to infection may demonstrate a link between mouse strain susceptibility, antibody production and specificity, and the ability to resolve an infection . The antibody response in five different strains of mice during primary immune response to systemic infection with Candida albicans was investigated . Immune sera were fractionated by protein A affinity chromatography to yield fractions containing IgG1, IgG2a and IgG2b immunoglobulins . BALB/c mice of low susceptibility to the infection and DBA/2J mice of high susceptibility produced increased levels of the IgG1 isotype and decreased levels of the IgG2a isotype . AKR, CBA/H and C57B1/6J mice of moderate susceptibility produced antibodies predominantly of the IgG2a isotype . The patterns of antigen recognition by antibodies in immune sera and in fractions obtained after protein A chromatography of immune sera were investigated by western blotting and immunostaining . Antibodies from AKR(H-2K) and CBA/H (H-2k) mice reacted strongly after immunoblotting with antigens of 87 and 96 kDa . In contrast, immune sera from both the highly susceptible DBA/2J (H-2d) mice and the resistant BALB/c (H-2d) mice reacted strongly with an antigen of 48 kDa . C57B1/6J (H-2b) mice produced variable antibody reactivity to antigens of 48, 65, 66 and 79 kDa depending on the IgG subclass tested . The IgG subclass responses and the patterns of antigen recognition in these mice suggest that humoral responses to C . albicans may be restricted by H-2 haplotype.(ABSTRACT TRUNCATED AT 250 WORDS)

J Clin Microbiol, 1995 Apr, 33(4), 962 - 7
Microtitration plate enzyme immunoassay to detect PCR-amplified DNA from Candida species in blood; Fujita S et al.; We developed a microtitration plate enzyme immunoassay to detect PCR-amplified DNA from Candida species . Nucleotide sequences derived from the internal transcribed spacer (ITS) region of fungal rDNA were used to develop species-specific oligonucleotide probes for Candida albicans, C . tropicalis, C . parapsilosis, and C . krusei . No cross-hybridization was detected with any other fungal, bacterial, or human DNAs tested . In contrast, a C . (Torulopsis) glabrata probe cross-reacted with Saccharomyces cerevisiae DNA but with no other DNAs tested . Genomic DNA purified from C . albicans blastoconidia suspended in blood was amplified by PCR with fungus-specific universal primers ITS3 and ITS4 . With the C . albicans-specific probe labeled with digoxigenin, a biotinylated capture probe, and streptavidin-coated microtitration plates, amplified DNA from a few as two C . albicans cells per 0.2 ml of blood could be detected by enzyme immunoassay.

J Clin Microbiol, 1995 Apr, 33(4), 915 - 7
Use of a colorimetric system for yeast susceptibility testing; Tiballi RN et al.; We examined the reliability and accuracy of a colorimetric assay using Alamar Blue reagent in the performance of susceptibility tests for Candida albicans . We compared the broth macrodilution method recommended by the National Committee for Clinical Laboratory Standards (NCCLS) with a macrodilution method modified with the Alamar reagent and a microdilution method modified with the Alamar reagent . The MICs of fluconazole and itraconazole for 97 isolates of C . albicans and 3 control isolates were tested . For fluconazole, the Alamar-modified broth macrodilution method yielded 94% (91 of 97) concordance within 2 dilutions compared with the NCCLS method, while the microdilution method yielded 95% (92 of 97) concordance . With Alamar-modified methods for itraconazole, broth macrodilution yielded 97% (94 of 97) concordance within 2 dilutions . MICs obtained by the microdilution method, although tightly nested, were shifted to a higher value when compared with those obtained by the NCCLS method; there was only 77% (75 of 97) concordance within 2 dilutions but 97% concordance (94 of 97) within 3 dilutions . Tests by all methods with quality control strains showed excellent reproducibilities . For fluconazole, the methods modified with the Alamar reagent yielded clear endpoints and excellent correlation for the broth macrodilution and microdilution methods . For itraconazole, the methods modified with the Alamar reagent yielded clear endpoints and were reproducible, but higher MICs were obtained by the microdilution methods compared with those obtained by the NCCLS methods.

J Clin Microbiol, 1995 Apr, 33(4), 794 - 6
Stability of karyotype in serial isolates of Candida albicans from neutropenic patients; Barton RC et al.; Serial isolates of Candida albicans were obtained from 20 patients with leukemia over periods of up to 8 months . The fingerprinting of these isolates by interrepeat PCR and random amplified polymorphic DNA PCR has been described previously (A . van Belkum, W . Melchers, B . E . de Pauw, S . Scherer, W . Quint, and J . F . Meis, J . Infect . Dis . 169:1062-1070, 1994) . Contour-clamped homogeneous electric field gel electrophoresis was used to examine the chromosomes of these isolates . When changes in the karyotype were seen in a series of isolates, additional interrepeat PCR and Southern blotting with a repeat DNA sequence from the 27A family were performed . These two genotyping tools were used to determine if karyotypic changes seen in a series of isolates were due to chromosome rearrangements in a single strain or due to colonization with more than one strain . It was determined that changes in karyotype in a series of strains indicated infection by a new strain.

Gesundheitswesen, 1995 Apr, 57(4), 214 - 22
{A new method for early detection of neurotoxic diseases (exemplified by pyrethroid poisoning)}; Muller-Mohnssen H et al.; This pilot-study should contribute to the question whether Pyrethroid intoxication can be distinguished from other diseases by characteristic clinical symptoms . The results show that the characteristics of the intoxication do not consist in singular symptoms but in combinations and correlations of symptoms, i.e . of central-neurological with peripheral- and autonomic-neurological as well as with characteristic immunological disturbances . Neurological symptoms consist in cerebro-organic disfunctions, locomotory disorders reminiscent of multiple sclerosis or M . Parkinson, and sensory, motoric and vegetative polyneuropathy, leading, for instance, to cardiovascular regulatory disorder like sympathicotonia or, orthostatic hypotonia . Non-neurological symptoms include immunosuppression with consecutive opportunistic infections, like candida albicans, most frequently of the alimentary tract, but also dermal and mucosal swellings, lichen-ruber-like efflorescences, loss of hair, conjunctivitis . Other symptoms are: hypoglycaemic crises inhibition of fertility, disturbances of blood clotting, and most frequently in children, suspected hematopoetic disorders.

Aust Dent J, 1995 Apr, 40(2), 91 - 7
Oral Candida albicans from patients infected with the human immunodeficiency virus and characterization of a genetically distinct subgroup of Candida albicans; McCullough M et al.; Candida albicans has been shown to vary in its phenotypic expression with the progression of human immunodeficiency virus (HIV) infection . This study was designed to investigate whether in Category IV HIV infected patients (CDC, Atlanta, USA) these phenotypic changes were related to changes in the genetic strain of the organism . Isolates of C . albicans were obtained from 45 patients with HIV infection during the progression of their disease as determined by percentage T4 lymphocyte count . Isolates were strain differentiated using two methods . In 67 per cent of the patients a single strain of C . albicans, as determined by the DNA analysis, was isolated from each individual over the experimental period . The phenotypic expression of the genetically identical strains isolated from each patient varied considerably over the experimental period with the morphotype 754 being predominant . These results showed that the genotype of C . albicans isolated persisted in the majority of HIV infected individuals, but that the phenotypical expression of this strain changed . A finding in this study was that 18 strains of C . albicans had DNA which did not hybridize to the probe used . These strains were analysed for the presence of two other C . albicans specific DNA segments using PCR . The probe 27A hybridizing strains yielded PCR products which differed from the non-hybridizing strains . Five of these genetically atypical C . albicans strains and 98 of the C . albicans strains were then analysed for purported virulence factors.(ABSTRACT TRUNCATED AT 250 WORDS)

Antimicrob Agents Chemother, 1995 Apr, 39(4), 958 - 63
Treatment of exogenous Candida endophthalmitis in rabbits with oral fluconazole; Park SS et al.; We investigated the efficacy of oral fluconazole, alone or in combination with oral flucytosine (5FC), in treating Candida endophthalmitis using a rabbit model . Albino rabbits were infected with an intravitreal inoculation of 1,000 CFU of susceptible Candida albicans and randomized 5 days later to receive treatment with oral fluconazole alone (80 mg/kg of body weight per day), a combination of fluconazole and 5FC (100 mg/kg/12 h), or no treatment . The treatment effect was assessed at 2 and 4 weeks after therapy by funduscopy, quantitative vitreous culture, and histopathology . Intravitreal levels of fluconazole, 2 to 24 h after the first dose, were measured to be > 10 times the MIC of the drug for C . albicans . Among rabbits treated with fluconazole for 2 weeks, 67% had a > 90% reduction in fungal load (P < 0.05) and 33% were sterile . After 4 weeks, all had a > 99% reduction in fungal load (P < 0.05) and 75% were sterile (P = 0.01) . This treatment effect was unchanged 4 weeks after discontinuation of fluconazole . Among rabbits treated with fluconazole and 5FC for 2 weeks, 67% died during therapy . Among the surviving rabbits, 75% had a > 90% reduction in fungal load (P < 0.05) and 25% were sterile . We conclude that oral fluconazole may be useful for treatment of Candida endophthalmitis . Addition of 5FC was associated with high toxicity and minimal additional antifungal effect in our rabbit model.

Antimicrob Agents Chemother, 1995 Apr, 39(4), 868 - 71
In vitro activity of D0870 compared with those of other azoles against fluconazole-resistant Candida spp; Wardle HM et al.; We compared the in vitro activity of a new triazole, D0870, with those of fluconazole, itraconazole, and ketoconazole against 41 clinical isolates of fluconazole-resistant Candida belonging to nine different species . The 50% inhibitory concentrations (IC50s) were determined by a microdilution method with morpholinopropanesulfonic acid (MOPS)-buffered RPMI medium and an inoculum of approximately 10(4) yeasts per ml . After incubation for 48 h at 37 degrees C the optical density at 550 nm was measured . The IC50 was the lowest drug concentration which reduced the optical density at 550 nm by > or = 50% compared with that for a drug-free control . D0870 had significant activity against many of the isolates . Its activity was comparable to that of ketoconazole, slightly superior to that of itraconazole, and markedly superior to that of fluconazole against Candida albicans . Against Candida glabrata, Candida krusei, and Candida inconspicua, it had activity similar to those of itraconazole and ketoconazole but had activity superior to that of fluconazole . D0870 IC50s for some isolates were increased . This may be due to cross-resistance mechanisms because the IC50s of both itraconazole and ketoconazole for these isolates were often high . When IC50s and IC80s were compared there was a marked organism and drug variation . With C . glabrata much higher endpoints for itraconazole were observed when an IC80 endpoint was used . For C . albicans there was also a significant shift upward in endpoints for itraconazole and ketoconazole . Values were changed little when IC50 and IC80 endpoints of D0870 were compared . For 35 of 41 isolates tested the D0870 IC50 was less than the 2.5-mg/liter breakpoint threshold proposed previously . Therefore, D0870 may be a useful agent for the therapy of infections caused by fluconazole-resistant Candida spp.

Enferm Infecc Microbiol Clin, 1995 Apr, 13(4), 209 - 12
{Assessment of the sensitivity to antifungal agents of clinical isolates of Candida albicans serotype A and B by the ATB Fungus method}; Quindos G et al.; BACKGROUND: Candida albicans is the most frequently species isolated in patients with candidiasis . This species may be antigenically divided in serotypes A and B, which have a different pathogenic behavior and antifungal susceptibility pattern . METHODS: The antifungal susceptibility of 443 clinical isolates from both serotypes of C . albicans to 5-fluorocytosine (5FC), amphotericin B, nystatin, econazole, miconazole and ketoconazole has been tested by means of the ATB Fungus method . RESULTS: Both serotypes showed a similar in vitro susceptibility to amphotericin B, nystatin, econazole, miconazole and ketoconazole . All the isolates were susceptible to amphotericin B and nystatin . A small number of in vitro resistant isolates were observed to azole compounds . However, serotype B was significantly more resistant than serotype A (p = 0.0001) to 5FC . CONCLUSIONS: C . albicans serotype B shows a lower susceptibility than serotype A to 5FC, an antifungal product not marketed in Spain . Serotyping of C . albicans does not offer additional information on antifungal susceptibility of clinical isolates from this species.

J Antibiot (Tokyo), 1995 Apr, 48(4), 306 - 10
A whole-cell Candida albicans assay for the detection of inhibitors towards fungal cell wall synthesis and assembly; Frost DJ et al.; A whole-cell C . albicans screen was designed to identify novel inhibitors interacting with the synthesis, assembly and regulation of the fungal cell wall . C . albicans was grown in a paired broth assay in 96-well plates with natural product extracts or pure chemical compounds in the presence and absence of the osmotic stabilizer, sorbitol . Growth was visually examined over a 7-day period and scored into different growth categories . Positives from the sorbitol rescue were then examined under the microscope for morphological alterations and grouped into several morphological classes . Sorbitol protection and cell morphology were indicators of novel antifungal agents from natural product extracts and pure compounds.

Clin Radiol, 1995 Apr, 50(4), 220 - 2
A histological basis for the 'sonographic snowstorm' in opportunistic infection of the liver and spleen; Keane MA et al.; We report 3 cases of opportunistic infection of the liver and spleen due to Pneumocystis carinii, Candida albicans and Aspergillus with an unusual but similar sonographic appearance . In the patient with Pneumocystis we report for the first time this same appearance in the bowel and pleura . Histology showed either extensive fibrosis or focal fibrinous exudates as the underlying cause . Calcification, though present, was scanty and was not thought to be the likely explanation for the appearances.

Am J Pathol, 1995 Apr, 146(4), 1017 - 24
Pulmonary alveolar proteinosis . A spontaneous and inducible disease in immunodeficient germ-free mice; Warner T et al.; Spontaneous pulmonary alveolar proteinosis (PAP), which resembles human PAP, was found in aging (35 to 40 weeks) germ-free SCID-beige (scid/scid-bg/bg) mice . Spontaneous PAP was not observed in germ-free SCID mice . We describe the induction of PAP in SCID mice monoassociated with a pure culture of Candida albicans for 15 to 40 weeks . The gastrointestinal tracts only are colonized, and disseminated or pulmonary candidiasis does not occur . Another spontaneous form of PAP, designated type II, was discovered in germ-free beige (bg/bg and bg/+) mice and in beige-nude (bg/bg-nu/nu) mice . In this form of PAP, macrophages appear to be unable to digest the ingested phospholipoprotein complex and then accumulate in the alveolar spaces . These murine models should prove useful in elucidating the relationships between immune deficiencies, infections, and cytokine regulation of granulocyte and macrophage production and function in pulmonary alveolar proteinosis.

J Med Microbiol, 1995 Apr, 42(4), 291 - 8
Various Candida and Torulopsis species differ in their ability to induce the production of C3, factor B and granulocyte-macrophage colony-stimulating factor (GM-CSF) in human monocyte cultures; Hogasen AK et al.; The incidence of infections with Candida albicans and also with non-albicans yeast species is increasing rapidly, particularly in immunocompromised patients . Eight Candida and Torulopsis species were compared for their ability to stimulate production of complement components C3 and factor B by monocytes . In addition, the release of granulocyte-macrophage colony-stimulating factor (GM-CSF) was determined, because this cytokine affects monocyte complement production . The highest ranked pathogenic yeasts, i.e., C . albicans, C . tropicalis and C . parapsilosis, were the most effective inducers of C3, factor B and GM-CSF production . C . krusei and T . glabrata showed intermediate activity, whereas C . kefyr, C . guilliermondii and T . candida had only a moderate stimulatory effect on C3 production and did not affect either factor B or GM-CSF release . The stimulated cytokine and complement production in response to the yeasts was highly variable in monocytes from different donors, but there was a consistent inverse relationship between C3 and GM-CSF concentrations in the monocyte supernates . This is in agreement with the previously described suppressive effect of GM-CSF on yeast-induced C3, but not factor B production . The monocyte responses elicited by a specific yeast species may be linked to its pathogenicity, and may also explain the predilection of some yeasts for particular underlying diseases.

Oral Microbiol Immunol, 1995 Apr, 10(2), 110 - 4
A novel method to study the hyphal phase of Candida albicans and to evaluate its hydrophobicity; Nikawa H et al.; As little is known about the surface properties of the hyphal phase of the dimorphic fungus Candida albicans due to the difficulty in examining the latter phase in isolation, a novel method was designed to compartmentalize the two phases of the yeast using a commercially available filter-type device-Chemotaxicell . When yeast cells in the blastospore phase were incubated in filter chambers of Chemotaxicell submerged in hyphal-induction media, hyphae traversed through the pores of the filter into the exterior of the chamber enmeshing the entire outer filter surface, after 72 h of incubation . However, the inner or the chamber surface of the filter comprised mainly blastospores . The hydrophobicity of the two morphologic phases of the yeast was then compared using this method, by contact angle measurement . When the 72-h specimens with matted hyphal elements were evaluated, the contact angles varied depending on the incubation medium (such as TC 199 or serum), and in each case hydrophobicity of the hyphal phase was significantly higher than the blastopore phase yeasts . This simple, reproducible method should help not only in evaluating the properties of the hyphal elements of C . albicans but also in studying parameters such as saliva and serum, which are known to affect hyphal formation in vivo.

Eur J Epidemiol, 1995 Apr, 11(2), 221 - 4
Comparison of seven phenotyping methods for Candida albicans; Otero L et al.; Seven different phenotyping methods for strain differentiation of Candida albicans (auxonotyping, enzymotyping, resistotyping, Phongpaichit's morphotyping, Hunter's morphotyping and Odds and Abbott's biotyping method-1980 and 1983 versions) were compared on a single population of 94 strains . 77.6% of the strains belonged to auxonotyping 1, 59.6% to enzymotyping A, 34% to resistotyping B and 30.8% to BC, 40.4% to Phongpaichit's morphotyping 000,000 and 40.4% to Hunter's morphotyping 'No fringe/Smooth surface' . Using biotyping systems (1980 and 1983 versions), the most frequent biotypes were 145 (29.8%) and 147 (31.9%) respectively . The Discriminatory Index of Hunter and Gaston was employed to carry out comparisons among the different systems . The best discriminatory results, although far from ideal, were found using Phongpaichit's morphotyping (DI = 0.827) and Odds and Abbott's method (DI = 0.815 and 0.831--1980 and 1983 versions) . A good discriminatory result was also found using Hunter's morphotyping method together with the biotyping of Odds and Abbott (1983 version) . These approximated the ideal (DI = 0.950) and showed minimal difficulty in interpretation . The proposed combined method revealed high discrimination among the vulvovaginal strains, and suggested the absence of transmissible pathogenic strains.

J Chemother, 1995 Apr, 7(2), 83 - 9
The activity of cilofungin on the incorporation of glucan associated proteins into hyphal cells of Candida albicans; Angiolella L et al.; The effect of the cilofungin, a beta 1-3 glucan synthase inhibitor, on the incorporation of the glucan associated proteins (GAP) into the mycelial wall of Candida albicans was investigated . For this study sub-inhibitory (< 2 micrograms/ml) doses of cilofungin were employed during the yeast to mycelial transition in a defined chemical medium, at 37 degrees C for 24 hours . Under these conditions, and particularly at the dose of 0.50 micrograms/ml cilofungin exerted a marked effect on GAP incorporation into the mycelial cell wall . The changes were essentially the absence of the two prominent bands of 46 and 31 kDa of the untreated cell wall coupled with an apparent increase in the amount of 55-56 kDa constituent, as well as of a minor constituent of 27-28 kDa . Radiolabel incorporation experiments demonstrated increased synthesis of a 34 kDa GAP, in addition to confirming the absence of the 46 kDa constituent, in mycelial cells under cilofunging treatment . Thus, sub-inhibitory doses of cilofungin may greatly alter the pattern of essential cell wall constituents such as the glucan-associated proteins, suggesting that this drug also has important effects on cell wall structure and fine organization, independent of, or prior to, its principal lytic effect on the fungal organism.

J Electron Microsc (Tokyo), 1995 Apr, 44(2), 72 - 8
Morphological aspects of cell wall formation during protoplast regeneration in Candida albicans; Nishiyama Y et al.; The dynamics of cell wall regeneration in Candida albicans protoplasts was investigated by fluorescence microscopy using a new fluorescent dye, Fungiflora Y, which specifically binds to beta-linked polysaccharides, as well as by electron microscopy . When freshly prepared protoplasts were incubated in osmotically stabilized medium, multiple binding sites of Fungiflora Y were detected on their surface within 15 min of incubation . At this initial stage of reversion, protoplasts began to form microfibrils outside the cytoplasmic membrane . From 1 h onward, the whole cell surface displayed a bright fluorescence . After 3 h of incubation, the reverting protoplasts were covered by an interwoven network of microfibrils . After 5 h of incubation, although most of the revertants had become osmotically resistant, regenerated cell walls in which the constituent materials appeared highly integrated were observed to be still monolayered in profile . Cell wall regeneration was complete after 24 h of incubation, at which time the reverted cells were capable of proliferating.

Br J Clin Pharmacol, 1995 Apr, 39(4), 460 - 2
The effect of amoxycillin on salivary nitrite concentrations: an important mechanism of adverse reactions?
Dougall HT, Smith L, Duncan C, Benjamin N.
Broad spectrum antibiotics are known to predispose towards oral candidiasis and gastroenteritis . Oral nitrite synthesis by commensal bacteria may be important in protecting the mouth and lower intestine from pathogenic organisms, including Candida albicans . The effect of 2 days administration of the broad spectrum antibiotic amoxycillin on salivary nitrite concentration, following a 200 mg potassium nitrate oral load, was studied in 10 healthy volunteers . The Cmax fell by 40% and the AUC was reduced by 1227 microM h (43%, 95% CI 273, 2181, P < 0.006) in the antibiotic treated group when compared with control . These findings suggest that destruction of nitrate reductase containing bacteria in the mouth by antibiotics may explain an increased incidence of infection with Candida and other pathogens.

J Antimicrob Chemother, 1995 Apr, 35(4), 509 - 19
Biodistribution of liposomal amphotericin B (AmBisome) and amphotericin B-desoxycholate (Fungizone) in uninfected immunocompetent mice and leucopenic mice infected with Candida albicans; van Etten EW et al.; The biodistribution of liposomal amphotericin B (L-AmB; AmBisome) and amphotericin B-desoxycholate were compared after a single injection of drug in uninfected immunocompetent mice and in leucopenic mice 6 h after inoculation with Candida albicans . Amphotericin B-desoxycholate was administered at the maximum tolerated dose (MTD) of 0.3 mg/kg whereas L-AmB was given at either 0.3 mg/kg or the MTD of 7 mg/kg . Amphotericin B (AmB) concentrations in the blood, liver, spleen, lungs and kidneys were determined by HPLC analysis at various intervals during the 48 h after administration . The biodistribution of both preparations of AmB followed similar patterns in both uninfected immunocompetent mice as well as those that were leucopenic and infected with C . albicans . Administration of L-AmB resulted in increased concentrations of drug in the blood, liver, and spleen but decreased concentrations in the kidney and lung . Hepatosplenic uptake of L-AmB was highly dose dependent with 7 mg/kg resulting in a relatively prolonged blood circulation . Blood and tissues retained high AmB concentrations after administration of L-AmB at the MTD . By using radiolabelled L-AmB, it was found that the high AmB concentrations in blood represented liposome-associated AmB and that during circulation in blood slow release of AmB occurred.

Res Commun Mol Pathol Pharmacol, 1995 Apr, 88(1), 115 - 8
Growth phase in relation to the lethal action of tamoxifen on Candida albicans; Beggs WH; Tamoxifen, a drug used to treat breast cancer, might have antifungal potential . The fungicidal activity of tamoxifen against Candida albicans surpassed that of the most active of the imidazole-type drugs . At 15-20 microM, tamoxifen rapidly killed yeast cells in stationary phase as well as in logarithmic phase, whereas imidazoles at this level generally do not exert significant lethal activity against stationary phase organisms.

Clin Exp Allergy, 1995 Apr, 25(4), 357 - 63
A standardized densitometric immunoblotting analysis of Candida albicans protein allergens; Savolainen J; Sera from 196 patients with a strong skin reactivity to Candida albicans and a history of atopic disease were used for C . albicans IgE-immunoblotting . The IgE-immunoblots were analysed by densitometry and integration of the densitogram peaks . A standardized reference disc system allowed parallel analysis of several different immunoblotting experiments . Altogether 105 patients had C . albicans-specific IgE antibodies in immunoblotting . The IgE-response against C . albicans was extremely polyspecific and was directed against 42 different bands . The most important IgE binding bands were 46 kDa and 27 kDa bands against which 44 (42%) and 29 (28%) patients, respectively, had IgE responses . A combination of IgE-immunoblotting and densitometry was found practical in analysis of large number of patient sera allowing a profound and accurate analysis and characterization of C . albicans allergen extract . This data evolved from a sufficiently large patient population can be utilized in further standardization of C . albicans extracts.

Clin Exp Allergy, 1995 Apr, 25(4), 350 - 6
Molecular cloning of cDNA coding for the 68 kDa allergen of Penicillium notatum using MoAbs; Shen HD et al.; To characterize the 68 kDa allergen of Penicillium notatum (also known as P . chrysogenum), a molecular antibody (MoAb) (P40) was previously generated . For cDNA cloning, three more MoAbs (3F, 5A3, 5G2) were generated in the present study . A mixture of all the four MoAbs was used in cloning of the gene coding for the 68 kDa allergen from a lambda gt11 cDNA library of P . chrysogenum . A cDNA clone (A6) with DNA insert of about 0.5 kb which encodes for the 3'-terminal nucleotide sequence of the 68 kDa allergen was obtained . The cloned sequence contained two putative N-glycosylation sites . The reduction in molecular weight from 68 to 62 kDa in immunoblotting after treatment of the crude extract of P . notatum with N-glycosidase F indicates that the 68 kDa allergen is a glycoprotein . Nucleotide sequence determination showed that 188 (54%) of the 348 nucleotides of the cDNA sequence obtained were identical to the same region of the nucleotide sequence of the beta-N-acetylglucosaminidase gene of Candida albicans . Although the cDNA clone obtained did not encode the full-length gene of the 68 kDa allergen, polypeptide expressed from the A6 cDNA showed positive immunological reactivities to all four MoAbs used in the cloning experiment and to IgE antibodies in sera of asthmatic patients . There was a loss of immunoblotting activity to the 68 kDa component after absorption of MoAb P40-containing culture supernatant with filters blotted on plaque lawns of cDNA clone A6 . Moreover, the immunoblotting activity remained when the MoAbs affinity-purified with filters containing polypeptides encoded by the cDNA insert of clone A6 were used.(ABSTRACT TRUNCATED AT 250 WORDS)

Arerugi, 1995 Apr, 44(4), 481 - 90
{Detection of IgE antibody to Pityrosporum orbiculare in patients with atopic dermatitis}; Ito K et al.; A lipophilic yeast, Pityrosporum orbiculare, found in normal microflora of the human skin, is considered to be a pathogenic allergen of atopic dermatitis (AD) . We measured IgE antibody to P . orbiculare using a CAP RAST FEIA kit in 44 patients with AD (AD alone 17, AD+bronchial asthma (BA) 27) . The incidence of positive RAST was 88.2% in the patients with AD alone and 74.1% in the patients with AD+BA . The IgE RAST index was 2.77 +/- 1.34 (mean +/- SD) in severe AD patients (n = 23), which was significantly higher than that in mild AD patients (1.88 +/- 1.41, n = 21) . The patients with eczematous skin on the face, neck and scalp (n = 27) showed higher IgE titers (2.79 +/- 1.26) than those without skin lesions (1.64 +/- 1.44, n = 17) . These results suggest that P . orbiculare is a pathogenic allergen of severe AD . IgE levels for P . orbiculare showed a statistically significant relation to those for Candida albicans (r = 0.62, p < 0.01) . In RAST inhibition tests on five patients' sera, C . albicans extract significantly inhibited IgE RAST to P . orbiculare in only one serum, indicating that P . orbiculare shares some IgE binding epitopes with C . albicans, but that P . orbiculare-specific epitopes also exist.

Mol Microbiol, 1995 Apr, 16(2), 241 - 50
Genetic studies reveal that myristoylCoA:protein N-myristoyltransferase is an essential enzyme in Candida albicans; Weinberg RA et al.; MyristoylCoA:protein N-myristoyltransferase (Nmt) catalyses the co-translational, covalent attachment of myristate (C14:0) to the amino-terminal glycine residue of a number of eukaryotic proteins involved in cellular growth and signal transduction . The NMT1 gene is essential for vegetative growth of Saccharomyces cerevisiae . Studies were carried out to determine if Nmt is also essential for vegetative growth of the pathogenic fungus Candida albicans . A strain of C . albicans was constructed in which one copy of NMT was partially deleted and disrupted . A Gly-447-->Asp mutation was introduced into the second NMT allele . This mutation produced marked reductions in catalytic efficiency at 24 and 37 degrees C, as judged by in vitro kinetic studies of the wild-type and mutant enzymes which had been expressed in, and purified from, Escherichia coli . The growth characteristics of isogenic NMT/NMT, NMT/delta nmt, and nmt delta/nmtG447D C . albicans strains were assessed under a variety of conditions . Only the nmt delta/nmtG447D strain required myristate for growth . This was true at both 24 and 37 degrees C . Palmitate could not substitute for myristate . Incubation of nmt delta/nmtG447D cells at 37 degrees C in the absence of myristate resulted in cell death as observed by the inability to form colonies on media supplemented with 500 microM myristate . Studies in an immunosuppressed-mouse model of C . albicans infection revealed that the NMT/delta nmt strain produced 100% lethality within 7 d after intravenous administration while the isogenic nmt delta/nmtG447G strain produced no deaths even after 21 d . These observations establish that Nmt is essential for vegetative growth of C . albicans and suggest that inhibitors of this acyltransferase may be therapeutically useful fungicidal agents.

Diagn Microbiol Infect Dis, 1995 Apr, 21(4), 191 - 4
Evidence of nosocomial spread of Candida albicans causing bloodstream infection in a neonatal intensive care unit; Reagan DR et al.; Candida albicans is an increasingly important bloodstream pathogen . We investigated a cluster of bloodstream infections in the neonatal intensive care unit (NICU) to determine whether nosocomial transmission occurred . Subjects included any patient in the NICU who developed clinically significant bloodstream infection with C . albicans from January 1984 to December 1987 (N = 7) . Isolates were typed by restriction fragment length polymorphism analysis using a C . albicans-specific DNA probe (27A) . Four of the neonates were infected from June to August 1984 (1.4 infections per 100 admissions) (the epidemic period) versus none in the period from January to May 1984, and three in the period from September 1984 to December 1987 (0.12 infections per 100 admissions) (P = .002) . Three of the four patients in the epidemic period were infected with identical strains, readily distinguished from epidemiologically unrelated strains from the NICU . We conclude that nosocomial transmission of C . albicans occurred and that neonates in intensive care units may represent one group at increased risk.

Cytometry, 1995 Apr 1, 19(4), 370 - 5
Enumeration of viable Candida albicans blastospores using tetrabromofluorescein (eosin Y) and flow cytometry; Costantino PJ et al.; A rapid assay was developed for determination of the viability of blastospores of Candida albicans utilizing flow cytometry to detect the accumulation of tetrabromofluorescein in non-viable yeast cells . Eosin Y was shown to stain non-viable C . albicans blastospores selectively without affecting the cellular viability of competent yeast cells or resulting in non-specific staining of viable cells . Results of these studies show that this flow cytometric method of determining cell viability of C . albicans is more accurate and more precise than the more common method of enumerating colony forming units.

Pol Arch Med Wewn, 1995 Apr, 93(4), 346 - 50
{Overdose of methotrexate with a fatal outcome in a patient with rheumatoid arthritis}; Porawska W; A 52-year old female patient with rheumatoid arthritis (RA) had mistakenly taken 112.5 mg methotrexate (MTX) over a period of five days . As a result, extensive erosions along with necrotic changes occurred in the oral cavity, the groins and vulva mucous membrane . Haemorrhages in the gastrointestinal tract, dermal xanthochromia, interstitial pneumonia and progressive renal insufficiency, as well as hair loss, also followed . Laboratory examination showed peripheral blood agranulocytosis, a decrease in the overall number of blood platelets, anaemia, an increase in the level of transaminases and bilirubin, along with megakaryocyte deficiency . Cultivated dermal specimens initially developed bacteria, then Candida albicans . The patient died on the 22nd day after the overdose from pneumonia and extensive mycosis.

J Ethnopharmacol, 1995 Apr, 46(1), 31 - 47
Screening of hundred Rwandese medicinal plants for antimicrobial and antiviral properties; Vlietinck AJ et al.; A series of 100 Rwandese medicinal plants (267 plant extracts), used by traditional healers to treat infections, were screened for antibacterial, antifungal and antiviral properties . The results of the testing showed that 45% were active against Staphylococcus aureus, 2% against Escherichia coli, 16% against Pseudomonas aeruginosa, 7% against Candida albicans, 80% against Microsporum canis and 60% against Trichophyton mentagrophytes . Not less than 27% of the plant species exhibited prominent antiviral properties against one or more test viruses, more specifically 12% against poliomyelitis, 16% against coxsackie, 3% against Semliki forest, 2% against measles and 8% against herpes simplex virus.

J Biol Chem, 1995 Mar 31, 270(13), 7272 - 80
Inositol trisphosphate-dependent and -independent Ca2+ mobilization pathways at the vacuolar membrane of Candida albicans; Calvert CM et al.; Vacuolar membrane vesicles were isolated from Candida albicans protoplasts, and marker enzyme assays were employed to identify the membranes as vacuolar in origin . The mechanisms of Ca2+ uptake and Ca2+ release at the vacuolar membrane were investigated . Ca2+ accumulation by vacuolar membrane vesicles can be generated via H+/Ca2+ antiport . The inside-acid pH is in turn generated by a vacuolar-type H(+)-ATPase, as demonstrated by the sensitivity of Ca2+ uptake to ionophores and the vacuolar H(+)-ATPase inhibitor bafilomycin A1 . Vacuolar membrane vesicles exhibit two Ca2+ release pathways: one induced by inositol 1,4,5-trisphosphate (InsP3) and the other by inside-positive voltage . These two pathways are distinct with respect to the amount of Ca2+ released, the nature of response to successive stimuli, and their respective pharmacological profiles . The InsP3-gated pathway exhibits a K0.5 for InsP3 of 2.4 microM but is not activated by inositol 4,5-bisphosphate or inositol 1,3,4,5-tetrakisphosphate at concentrations up to 50 microM . Ca2+ release by InsP3 is blocked partially by low molecular weight heparin . Ca2+ released by the voltage-sensitive pathway occurs at membrane potentials estimated to be over a physiological range from 0 to 80 mV . The voltage-sensitive Ca2+ release pathway can be blocked by lanthanide ions and organic channel blockers such as ruthenium red and verapamil . Furthermore, the voltage-sensitive Ca2+ release pathway exhibits Ca(2+)-induced Ca2+ release . These findings are discussed in relation to the mechanism of Ca(2+)-mediated cellular signaling in C . albicans and other fungi.

Proc Natl Acad Sci U S A, 1995 Mar 28, 92(7), 2544 - 8
Molecular cloning and characterization of chitinase genes from Candida albicans; McCreath KJ et al.; Chitinase (EC 3.2.1.14) is an important enzyme for the remodeling of chitin in the cell wall of fungi . We have cloned three chitinase genes (CHT1, CHT2, and CHT3) from the dimorphic human pathogen Candida albicans . CHT2 and CHT3 have been sequenced in full and their primary structures have been analyzed: CHT2 encodes a protein of 583 aa with a predicted size of 60.8 kDa; CHT3 encodes a protein of 567 aa with a predicted size of 60 kDa . All three genes show striking similarity to other chitinase genes in the literature, especially in the proposed catalytic domain . Transcription of CHT2 and CHT3 was greater when C . albicans was grown in a yeast phase as compared to a mycelial phase . A transcript of CHT1 could not be detected in either growth condition.

J Heart Valve Dis, 1995 Mar, 4(2), 171 - 5
Porcine stentless aortic valve in re-replacements and acute aortic valve endocarditis; Gontijo BF et al.; Between June 1990 and June 1993, 135 patients received a porcine stentless aortic valve (PSAV) at our institution . In this group, there were 33 patients in whom the stentless valve was used to replace a previously inserted malfunctioning aortic valve prosthesis (n = 19) or to treat acute aortic valve endocarditis (n = 14, five native, nine prosthetic endocarditis) . There was one hospital death caused by multiple organ failure in a patient with endocarditis and preoperative cerebral stroke . Two patients died after hospital discharge; one suffered sudden death following a pacemaker failure four month after surgery and the other died due to Candida albicans sepsis after a prolonged antibiotic treatment . There were two reoperations; one to correct a dehiscence of the proximal suture line and the other to replace a degenerated valve 3.5 years after implantation in a 14 year old boy . All operative survivors were followed clinically with serial color Doppler echocardiography . No recurrence of endocarditis was detected . Aortic root reconstruction was achieved even in the presence of multiple abscesses . All but one patients showed a normally functioning valve with none or minimal aortic insufficiency . In our opinion the PSAV is an excellent aortic valve substitute for patients with damaged aortic annulus, because it promotes aortic root remodeling, decreases the incidence of postoperative paravalvular leaks and helps to prevent endocarditis recurrence.

J Med Microbiol, 1995 Mar, 42(3), 225 - 9
Non-opsonic phagocytosis of Trichophyton mentagrophytes arthroconidia by human neutrophils in vitro; Richardson MD et al.; A non-opsonic mechanism of binding and phagocytosis by human neutrophils of Trichophyton mentagrophytes arthroconidia is described . This was in direct contrast to the complement dependency of Candida albicans phagocytosis . Both serum complement and specific antibody to T . mentagrophytes promoted maximal phagocytosis (61% and 40% of neutrophils, respectively, contained arthroconidia) . Increasing the ratio of arthroconidia to neutrophils did not increase non-opsonic phagocytosis (18-26%) . Phagocytosis of arthroconidia exposed to trypsin in the absence of opsonin was not affected (18%) . However, proteinase and chitinase reduced the level of non-opsonic and opsonic phagocytosis to negligible levels (6.3% and 4.5%, respectively) . When mannose was added to neutrophils, mannose receptors on the phagocyte membrane were partially blocked when arthroconidia were opsonised, but this did not reduce the level of non-opsonic phagocytosis . The non-opsonic mechanism proposed here may have direct relevance in skin sites poor in opsonins.

J Am Acad Dermatol, 1995 Mar, 32(3), 429 - 35
Antifungal activity of itraconazole and terbinafine in human stratum corneum: a comparative study; Pierard GE et al.; BACKGROUND: The evaluation of antifungal agents by in vitro and animal experiments cannot predict clinical efficacy with certainty . New models are needed to assess and compare antifungal activity . OBJECTIVE: We compared on human stratum corneum ex vivo the antifungal activity and lingering effect of 200 mg itraconazole daily and twice daily, and 250 mg/day terbinafine . METHODS: Three groups of 10 healthy volunteers entered the open comparative trial . Results were evaluated in a blinded manner . Cyanoacrylate skin surface strippings (CSSS) were taken from the back and superficial dermatome skin samples (SDSS) were taken from plantar skin at days 0, 1, 3, 7, 8, 10, 14, 21, 28, and 35 . Spores or yeasts of selected fungi (Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, and Candida albicans) were deposited and cultured on the CSSS and SDSS . The 1-week fungal growth on CSSS and SDSS was assessed over time by computerized image analysis to derive the inhibitory effect of the oral antifungal agents administered . Fungitoxic activity was also assessed by the use of 2-day cultures on CSSS followed by a transfer to Sabouraud medium . RESULTS: Comparable antifungal activity against dermatophytes was found for all three regimens . Itraconazole at both dosages was always significantly more active than terbinafine against C . albicans on CSSS and SDSS . Overall, 200 mg itraconazole twice daily appeared to be more fungitoxic than 250 mg/day terbinafine and 200 mg/day itraconazole . CONCLUSION: The ex vivo culture of fungi on human stratum corneum is very similar to the in vivo situation . Both itraconazole and terbinafine display high antidermatophyte activity . Faster onset and longer posttherapy activity were demonstrated in the itraconazole treatment groups . Terbinafine had marginal activity against C . albicans in this model.

J Bacteriol, 1995 Mar, 177(5), 1239 - 46
Structure and regulation of a Candida albicans RP10 gene which encodes an immunogenic protein homologous to Saccharomyces cerevisiae ribosomal protein 10; Swoboda RK et al.; The Candida albicans clone cDNA10 was isolated on the basis that it encodes a protein which is immunogenic during infections in humans (R . K . Swoboda, G . Bertram, H . Hollander, D . Greenspan, J . S . Greenspan, N . A . R . Gow, G . W . Gooday, and A . J . P . Brown, Infect . Immun . 61:4263-4271, 1993) . cDNA10 was used to isolate its cognate gene, and both the cDNA and gene were sequenced, revealing a major open reading frame with the potential to encode a basic protein of 256 amino acids with a predicted molecular weight of 29 kDa . Over its entire length, the open reading frame showed strong homology at both the nucleic acid (75 to 78%) and amino acid (79 to 81%) levels to two Saccharomyces cerevisiae genes encoding the 40S ribosomal protein, Rp10 . Therefore, our C . albicans gene was renamed RP10 . Northern (RNA) analyses in C . albicans 3153 revealed that RP10 expression is regulated in a manner very similar to that of S . cerevisiae ribosomal genes . The level of the RP10 mRNA decreased upon heat shock (from 25 to 45 degrees C) and was tightly regulated during growth . Maximal levels of the mRNA were reached during mid-exponential phase before they decreased to negligible levels in stationary phase . The level of the RP10 mRNA was induced only transiently during the yeast-to-hyphal morphological transition but did not appear to respond to hyphal development per se.

Infect Immun, 1995 Mar, 63(3), 984 - 8
Degradation of humoral host defense by Candida albicans proteinase; Kaminishi H et al.; The effect of an extracellular proteinase from the pathogenic yeast Candida albicans on the bactericidal and opsonizing activities of human serum was studied . The ability of human polymorphonuclear leukocytes to kill Staphylococcus aureus was greatly reduced when the bacteria were opsonized with human serum treated with the proteinase . The reduction in the opsonizing activity of human serum was attributed to degradation of the Fc portion of immunoglobulin G by the action of C . albicans proteinase as determined by immunoprecipitation reaction . However, the Fab portion of immunoglobulin G was resistant to proteolysis by the proteinase . A clear reduction in the bactericidal activity of human serum against Escherichia coli was observed when the serum was treated with C . albicans proteinase . The reduction of serum bactericidal activity was attributed to the degradation of complement C3 by proteolysis by the proteinase as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, while C5 resisted the action of the proteinase . As determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the proteinase also degrades endogenous proteinase inhibitors, such as alpha 2 macroglobulin and alpha 1 proteinase inhibitor, which are involved in regulating inflammation . These results suggest that destruction of a host's defense-oriented or regulatory proteins facilitates debilitation of the infected host.

Infect Immun, 1995 Mar, 63(3), 1142 - 4
Nitric oxide production does not directly increase macrophage candidacidal activity; Vazquez-Torres A et al.; Some activated murine macrophages produced nitrite but were unable to kill Candida albicans . Furthermore, a nitric oxide (NO) generator inhibited C . albicans growth but was not candidacidal . Our results suggest that NO is candidastatic and that No is not directly involved but is associated with or induces other macrophage candidacidal mechanisms.

Mol Cell Biol, 1995 Mar, 15(3), 1797 - 805
Functional analysis of the promoter of the phase-specific WH11 gene of Candida albicans; Srikantha T et al.; Candida albicans WO-1 switches spontaneously, frequently, and reversibly between a hemispherical white and a flat gray (opaque) colony-forming phenotype . This transition affects a number of morphological and physiological parameters and involves the activation and deactivation of phase-specific genes . The WH11 gene is transcribed in the white but not the opaque phase . A chimeric WH11-firefly luciferase gene containing the 5' upstream region of WH11 was demonstrated to be under phase regulation regardless of the site of integration, and a series of promoter deletion constructs was used to delineate two white-phase-specific transcription activation domains . Gel retardation experiments with the individual distal or proximal domain and white-phase or opaque-phase protein extract demonstrated the formation of one distal white-phase-specific complex and two proximal white-phase-specific complexes . Specific subfragments were tested for their ability to compete with the entire domain in the formation of complexes with white-phase protein extract in order to map the proximal domain sequence involved in white-phase-specific complex formation . Our results indicate that white-phase-specific transcription of WH11 is positively regulated by trans-acting factors interacting with two cis-acting activation sequences in the WH11 promoter.

Antimicrob Agents Chemother, 1995 Mar, 39(3), 598 - 601
Combination therapy of murine invasive candidiasis with fluconazole and amphotericin B; Sugar AM et al.; A study was performed to assess the in vivo relevance of the in vitro antagonism between fluconazole and amphotericin B against Candida albicans . Combinations of fluconazole and amphotericin B were explored for their efficacies against acute (100% mortality in 2 to 5 days) or less acute (100% mortality in 30 days) invasive candidiasis infections in mice with healthy immune systems and immunocompromised mice . Treatment efficacy was assessed by protection from mortality and/or a reduction in the fungal burden in tissue . In models of acute infection in mice with healthy immune systems or less acute infection in immunocompromised mice, combinations of fluconazole and amphotericin B were superior to fluconazole alone, and the effects were at least additive . Combination therapy was at least as efficacious as amphotericin B alone . In a different model of less acute infection in mice with healthy immune systems, combinations of fluconazole and amphotericin B showed no interactions and were no better than either drug alone . We conclude that combination therapy with fluconazole and amphotericin B is not antagonistic in vivo, in contrast to published in vitro studies, and, consequently, suggest that combination therapy should be considered in the management of clinical candidiasis.

C R Acad Sci III, 1995 Mar, 318(3), 359 - 65
Mycobacterial polar glycopeptidolipids enhance resistance to experimental murine candidiasis; Lagrange PH et al.; Intraperitoneal administration of polar glycopeptidolipids extracted from Mycobacterium chelonae (pGPL-Mc) greatly increased the resistance of mice against a lethal disseminated Candida albicans infection . This enhanced resistance was demonstrated by an increase in the number of survivors and the prolongation of the mean survival time of animals following a lethal challenge . These effects were dependent upon the infective dose of Candida albicans, the dose of pGPL-Mc and the timing of its administration . This enhanced resistance was correlated with the development and persistence of a hyperleukocytosis, associated with a long lasting increase in the number of polymorphonuclear neutrophils . On the contrary, no candidacidal effect of the serum collected from pretreated mice was observed; suggesting that the ability of pGPL-Mc to increase resistance against Candida albicans infection is likely to be mediated by polymorphonuclear neutrophils . These results confirm previously described immunostimulating properties of pGPL-Mc and open the way for the evaluation of its effect in the prevention of opportunistic infections in neutropenic patients.

Cell Mol Biol (Noisy-le-grand), 1995 Mar, 41(2), 297 - 305
Internalization of Candida albicans and cytoskeletal organization in macrophages and fibroblasts treated with concanavalin A; Bodo M et al.; This paper investigates the ability of macrophages and of non-typically phagocitic cells such as fibroblasts to internalize 51Cr-labelled C . albicans in presence or in absence of lectin concanavalin A (Con A) . The results obtained demonstrate that fibroblasts are also able to internalize C . albicans and that this property is potentiated by the presence of Con A . Lectin modifies only the phenotype of the fibroblast, which, poorly attached to the substrate, is globular in shape . Despite reduced cellular spreading, phagocytosis is stimulated by the lectin . In both cell populations, changes in the organization of some cytoskeletal proteins such as tubulin, actin and alpha-actinin are evident during the C . albicans infection; such rearrangements are more evident and longlasting in the fibroblasts treated with Con A.

Chem Pharm Bull (Tokyo), 1995 Mar, 43(3), 441 - 9
Optically active antifungal azoles . V . Synthesis and antifungal activity of stereoisomers of 3-azolyl-2-(substituted phenyl)-1-(1H-1,2,4-triazol-1-yl)-2- butanols; Tasaka A et al.; The (2S,3S)-, (2R,3S)- and (2S,3R)-stereoisomers of (2R,3R)-3-azolyl-2-(substituted phenyl)-1-(1H-1,2,4-triazol-1-yl)-2-butanols {(2R,3R)-1a--d} were prepared and evaluated for antifungal activity against Candida albicans in vitro and in vivo to clarify the relationships between stereochemistry and biological activities . The results revealed that the in vitro antifungal activity in each set of the four stereoisomers {(2R,3R)-, (2S,3S)-, (2R,3S)- and (2S,3R)-1a--d} definitely paralleled the in vivo antifungal activity against candidosis in mice, and the order of potency was (2R,3R) >> (2R,3S) > or = (2S,3S) > or = (2S,3R) . In addition, the four stereoisomers in each set were assessed for sterol biosynthesis-inhibitory activities in C . albicans and rat liver . The (2R,3R)-isomer was found to exert a strong and selective inhibitory effect on the sterol synthesis in C . albicans as compared with that in rat liver.

Chem Pharm Bull (Tokyo), 1995 Mar, 43(3), 432 - 40
Optically active antifungal azoles . IV . Synthesis and antifungal activity of (2R,3R)-3-azolyl-2-(substituted phenyl)-1-(1H-1,2,4-triazol-1-yl)-2-butanols; Tasaka A et al.; (2R,3R)-3-Azolyl-2-(substituted phenyl)-1-(1H-1,2,4-triazol-1-yl)-2-butanols (III) were prepared from (2R,3S)-3-methyl-2-(substituted phenyl)-2-(1H-1,2,4-triazol-1-yl)methyloxiranes (21a-f) by a ring-opening reaction with 1H-1,2,3-triazole and 1H-tetrazole and evaluated for antifungal activity against Candida albicans in vitro and in vivo . The optically active oxiranes (21a--f) which serve as the key synthetic intermediates, were synthesized from 1-{(2R)-2-(3,4,5,6-tetrahydro-2H-pyran-2-yl)oxypropanoyl}morpholin e (24) and substituted phenylmagnesium bromide (23) via six steps in a stereocontrolled manner . The 3-(1H-1,2,3,-triazol-1-yl)-(IIIa) and 3-(2H-2-tetrazolyl)-2-butanol (IIId) derivatives showed strong protective effects against candidosis in mice.

J Can Dent Assoc, 1995 Mar, 61(3), 199 - 200, 203-5
Effectiveness of a topical antifungal regimen for the treatment of oral candidiasis in older, chronically ill, institutionalized, adults; Banting DW et al.; Because of predisposing systemic disease, the frequent administration of medication, and the use of a complete denture, oral candidiasis is a common problem among older, chronically ill, institutionalized adults . This randomized clinical trial was designed to evaluate the effectiveness of an antifungal denture soaking solution (48 mL nystatin liquid, 100,000 IU/mL, dissolved in 432 mL of distilled water producing 10,000 IU nystatin mL solution), used as an adjunct to a nystatin vaginal lozenge (100,000 IU/g, dissolved in the mouth three times daily for seven days) in a group of older, chronically ill, institutionalized adults . Although the clinical signs and symptoms of oral candidiasis were resolved in all subjects following therapy, the presence of invasive Candida hyphae was detected in approximately 80 per cent of tissue and/or dentures . When compared to tap water, the use of an antifungal denture soaking solution produced no detectable difference in the presence of Candida albicans hyphae over a three-month period (M-H chi-square = 0.021, p = 0.886), but it did reduce the rate of recurrence of clinical signs and symptoms . The appropriateness of this regimen for the treatment of oral candidiasis in this type of patient is challenged.

Scand J Gastroenterol, 1995 Mar, 30(3), 216 - 8
Omeprazole and dry mouth; Teare JP et al.; BACKGROUND: Omeprazole causes irreversible inhibition of the hydrogen/potassium adenosine triphosphatase enzyme, leading to a marked reduction in both acid secretion and volume of gastric juice . Reported side-effects include nausea, vomiting, diarrhoea, constipation, and headache . We report the development of dry mouth during omeprazole therapy . METHODS: We have identified six patients taking omeprazole for more than 6 weeks who complained of a dry mouth . Salivary production was measured as whole salivary flow produced over a 10-min period spat into a collecting vessel and as 5% citric acid-stimulated parotid salivary flow collected with a Lashley cup device placed over the parotid duct . Flow rates were evaluated both during and after cessation of treatment . Saliva produced was then cultured for microbes . RESULTS: Four of the six had subnormal parotid or whole salivary flow rates on treatment that recovered after stopping treatment . The increase after treatment was marked in four . Significant amounts of Candida albicans grew from the saliva of the three patients with the lowest salivary flows; one saliva also grew Staphylococcus aureus . CONCLUSION: Salivary flow is reduced in some patients treated with omeprazole, returning to normal after cessation of treatment . This reduction may predispose to opportunistic infection, particularly in the edentulous.

Epidemiol Mikrobiol Imunol, 1995 Mar, 44(1), 33 - 5
{Secretory proteases of pathogenic Candida}; Kotyza J et al.; The enzymic activity of secreted aspartic proteinase (SAP) in 117 clinical isolates of Candida albicans strains ranges from 0.0 do 0.12 mumol Tyr/min/ml of culture supernatants following 24 h incubation in a YCB-BSA-glucose medium . SAP was purified by the modification of a combined chromatographic method (Banerjee et al., 1991) and a final gel filtration . The diagnostic value of the SAP activity estimation and immunochemical use of purified enzyme are discussed.

Clin Infect Dis, 1995 Mar, 20(3), 634 - 40
DNA subtypes and fluconazole susceptibilities of Candida albicans isolates from the oral cavities of patients with AIDS; Barchiesi F et al.; Sixty-two Candida albicans isolates from the oral cavities of 28 patients with AIDS who were receiving fluconazole therapy were typed by restriction endonuclease analysis followed by pulsed-field gel electrophoresis; these isolates were then tested for fluconazole susceptibility by a standard broth dilution method . Sequential isolates (range, 2-4) were evaluated for 22 patients; only one isolate was evaluated for six patients . DNA subtyping revealed a total of 37 different DNA subtypes . Twelve (54.5%) of 22 patients with multiple episodes of oropharyngeal candidiasis were infected with a single DNA subtype throughout the observation period . Ten (45.5%) of 22 patients with multiple episodes of oropharyngeal candidiasis were infected with two or three DNA subtypes during the observation period . In vitro susceptibility tests revealed that MICs of fluconazole ranged from < or = 0.125 microgram/mL to 64 micrograms/mL, with an MIC50 of 0.5 microgram/mL and an MIC90 of 4 micrograms/mL . A significant increase in the MICs (fourfold or greater) of fluconazole for sequential C . albicans isolates was found for 66.6% of the patients infected with a single DNA subtype and for 50% of the patients infected with multiple DNA subtypes . Despite a limited number of patients and isolates, our data suggest that C . albicans isolates that are susceptible to fluconazole at MICs of > or = 8 micrograms/mL in vitro will be less susceptible in vivo to standard doses (100-200 mg/d) of this drug.

J Clin Microbiol, 1995 Mar, 33(3), 769 - 71
Molecular epidemiology of Candida isolates from AIDS patients showing different fluconazole resistance profiles; Lischewski A et al.; Thirty Candida isolates obtained from the oropharynxes of three AIDS patients were genotypically characterized . In vitro fluconazole MIC determination revealed increasing fluconazole resistances during treatment, thereby confirming the in vivo situation . Pulsed-field gel electrophoresis karyotyping, randomly amplified polymorphic DNA analysis, and hybridizations with Candida albicans repetitive element 2 were used to determine possible genotypic changes . The isolates from two patients showed genetic homogeneity, suggesting the selection for resistant variants . One patient experienced a strain switch to Candida krusei . Horizontal spread of identical strains between the patients could be excluded . However, the molecular methods used might not be sufficient to detect the underlying genetic basis of resistance to fluconazole.

J Clin Microbiol, 1995 Mar, 33(3), 696 - 700
Characterization of genetically distinct subgroup of Candida albicans strains isolated from oral cavities of patients infected with human immunodeficiency virus; McCullough M et al.; During the course of a study of oral Candida albicans strains from 60 human immunodeficiency virus-infected patients over a 2.5-year period, 18 of the 295 C . albicans isolates had genomes that failed to hybridize with a C . albicans-specific DNA probe (27A) . These strains were germ tube positive and chlamydospore positive and were identified as C . albicans by the ID 32C test (API Systems, Montlieu, France) . These strains were analyzed for the presence of two other C . albicans-specific DNA segments by PCR . The first was a C . albicans 1,348-bp species-specific sequence, and the second was a 1,059-bp C . albicans repetitive element . The probe 27A-hybridizing strains yielded PCR products which differed from those of the nonhybridizing strains . Five of these genetically atypical C . albicans strains and 98 of the C . albicans strains were then analyzed for purported virulence factors . The genetically atypical C . albicans strains, in comparison with typical C . albicans strains, produced greater amounts of extracellular proteinase (P = 0.038, Student's t test), adhered to a greater degree to buccal epithelial cells (P = 0.018, Student's t test), and were less susceptible to the antifungal drug 5-flucytosine (P = 0.0003, Mann-Whitney test) . Analysis of these strains with other common antifungal drugs showed no statistically significant variation in susceptibility . The results of this study indicated that these genetically atypical C . albicans strains possess increased virulence in comparison with typical C . albicans strains.

J Clin Microbiol, 1995 Mar, 33(3), 661 - 7
Susceptibility testing of Candida albicans and Aspergillus species by a simple microtiter menadione-augmented 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay; Jahn B et al.; We describe a simple microtiter method for determining the susceptibility of Candida albicans and hyphal forms of Aspergillus fumigatus against antifungal agents . The assay measures mitochondrial respiration by determining reduction of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to formazan, a process that is enhanced in the presence of menadione . C . albicans or conidial suspensions of A . fumigatus are seeded into microtiter plates . Hyphal outgrowth of Aspergillus spp . was achieved by a 12 to 14-h culture at 30 degrees C . Antifungal agents (amphotericin B, fluconazole, itraconazole) were added to the cultures for 24 h . Thereafter, incubations were continued for 3 h in the presence of MTT plus 0.1 mM menadione . Formazan formation was quantified photometrically after extraction of the formazan with acid isopropanol . Well-defined dose-response curves reflecting impairment of mitochondrial function by the antifungal agents were obtained . With C . albicans, the results correlated excellently with the MIC determinations performed according to the standard macrodilution procedure . In confirmation of a recent report, it was found that fluconazole was unable to exert its fungistatic action on a sensitive C . albicans strain in the presence of serum . The presented method can easily be integrated in the standard repertoire of a diagnostic microbiology laboratory and should prove useful as a means to assess the antifungal action of various agents on yeasts and filamentous fungi in the presence and absence of serum proteins or body fluids.

J Clin Microbiol, 1995 Mar, 33(3), 625 - 8
Improved detection of Candida albicans by PCR in blood of neutropenic mice with systemic candidiasis; van Deventer AJ et al.; A PCR using primers aimed at the multicopy gene coding for the small subunit rRNA and resulting in the synthesis of a 180-bp fragment was evaluated for its use in diagnosing invasive candidiasis in comparison with blood culture . With the use of a C . albicans-specific probe, +/- 10 to 15 C . albicans cells are detected in 100 microliters of whole blood by Southern analysis . A DNase pretreatment was critical in the purification process of yeast DNA from whole blood . Omission of the DNase pretreatment decreased assay sensitivity 10-fold . PCR analysis of blood specimens collected from mice with invasive candidiasis is more sensitive than blood culture (100 versus 67%, respectively) at 72 h after intravenous (i.v.) inoculation with C . albicans . Furthermore, the intensity of the hybridization signals increased with the progression of infection . In contrast, multiple blood samples from gastrointestinally colonized mice were all negative by PCR, indicating that the PCR assay is also specific and may, therefore, make a positive contribution to the detection and follow-up of invasive candidiasis.

J Clin Microbiol, 1995 Mar, 33(3), 576 - 80
Investigation of Candida albicans transmission in a surgical intensive care unit cluster by using genomic DNA typing methods; Voss A et al.; An apparent outbreak of serious Candida albicans infections (n = 6) occurred in a surgical intensive care unit over a 4-week period . Four patients developed C . albicans bloodstream infections . An additional patient developed catheter-related C . albicans infection; the sixth patient developed an infection of cerebrospinal fluid . C . albicans was isolated from the hands of five health care workers (17%) and the throat of one health care worker (3%) during the outbreak investigation . Karyotyping and restriction endonuclease analysis of genomic DNA with BssHII of 23 C . albicans isolates from patients and the 6 health care worker isolates revealed 9 and 12 different patterns, respectively . Three of six patients appeared to be infected with the same C . albicans strain (two bloodstream infections and one cerebrospinal fluid infection) . The hands of a health care worker were colonized with strain that appeared identical to an isolate from a patient prior to infection of the patient . However, restriction endonuclease analysis with SfiI found differences among the isolates determined to be identical by the other two methods . Karyotyping alone does not appear to be sufficient to differentiate between outbreak and control isolates . Restriction endonuclease analysis typing may be a more sensitive method than karyotyping alone in the investigation of a cluster of C . albicans infections . Furthermore, the use of more than one restriction enzyme may be necessary for optimal strain discrimination in restriction endonuclease analysis of genomic DNA.

J Appl Bacteriol, 1995 Mar, 78(3), 264 - 9
Antimicrobial activity of the major components of the essential oil of Melaleuca alternifolia; Carson CF et al.; Tea tree oil, or the essential oil of Melaleuca alternifolia, is becoming increasingly popular as a naturally occurring antimicrobial agent . The antimicrobial activity of eight components of tea tree oil was evaluated using disc diffusion and broth microdilution methods . Attempts were also made to overcome methodological problems encountered with testing compounds which have limited solubility in aqueous media . After assessing media with and without solubilizing agents, the disc diffusion method was used to determine the susceptibility of a range of micro-organisms to 1,8-cineole, 1-terpinen-4-ol, rho-cymene, linalool, alpha-terpinene, gamma-terpinene, alpha-terpineol and terpinolene . While the disc diffusion method lacked reproducibility, it was considered useful as a procedure for screening for antimicrobial activity . Terpinen-4-ol was active against all the test organisms while rho-cymene demonstrated no antimicrobial activity . Linalool and alpha-terpineol were active against all organisms with the exception of Pseudomonas aeruginosa . Minimum inhibitory and minimum cidal concentrations of each component against Candida albicans, Escherichia coli and Staphylococcus aureus were determined using a broth microdilution method . Modifications to this method overcame solubility and turbidity problems associated with the oil components and allowed the antimicrobial activity of each of the components to be quantified reproducibly . There was reasonable agreement between minimum inhibitory concentrations and zones of inhibition . These results may have significant implications for the future development of tea tree oil as an antimicrobial agent.

J Med Vet Mycol, 1995 Mar-Apr, 33(2), 131 - 6
Effectiveness of fluconazole in murine Candida albicans and bacterial C . albicans peritonitis and abscess formation; Sawyer RG et al.; The role of fluconazole in the treatment of many forms of focal mycoses remains unclear . We studied the effectiveness of three different oral doses of fluconazole in three murine models of Candida albicans peritonitis leading to intra-abdominal abscess formation . During monomicrobial Candida infection, fluconazole decreased mortality and the number of C . albicans cultured per abscess; prolonged treatment also eliminated Escherichia coli translocation . In mixed C . albicans/E . coli/Bacteroides fragilis infection, prolonged treatment with higher doses of fluconazole decreased mortality, the number of abscesses formed, and the number C . albicans per abscess . In animals with a similar polymicrobial infection but with concurrent cefoxitin treatment, fluconazole decreased mortality and the number of C . albicans per abscess; in addition, prolonged treatment reduced the number of abscesses . Amphotericin B gave similar results in all three models . These data indicate that the clinical use of fluconazole in peritonitis should be investigated.

J Med Vet Mycol, 1995 Mar-Apr, 33(2), 127 - 30
Fungal cultures on cyanoacrylate skin surface strippings as a dose-finding method for topical antifungals . A placebo-controlled study with 0.25% and 0.50% itraconazole cream; Arrese JE et al.; The antimycotic activities of 0.25% and 0.50% itraconazole cream were compared in the stratum corneum after once-daily applications for 1 week . Two groups of 12 healthy volunteers applied either itraconazole or placebo on the inner side of each forearm, in a double-blind design . Cyanoacrylate skin surface strippings (CSSS) were taken on days 8, 11 and 21 . Conidia or yeasts of selected fungi (Trichophyton rubrum, Trichophyton metagrophytes, Microsporum canis and Candida albicans) were deposited on CSSS . Fungal growth on CSSS was assessed in time by computerized image analysis to derive the inhibitory effect of the previously applied antifungal preparations . Comparable antimycotic activity was found against dermatophytes for both concentrations . Itraconazole 0.50% appeared to be more active than 0.25% against C . albicans . The 0.50% concentration yielded prominent fungitoxic effect after 1 week of treatment, and showed a lingering effect in the stratum corneum for at least 3 days . This method could be useful in a pre-clinical setting and serve as a predictive tool for further clinical dose-finding studies with topical antimycotics.

J Med Vet Mycol, 1995 Mar-Apr, 33(2), 117 - 22
Influence of cell surface hydrophobicity on attachment of Candida albicans to extracellular matrix proteins; Silva TM et al.; Cell surface hydrophobicity expression by Candida albicans facilitates a diffuse binding distribution of yeast cells to host tissues ex vivo . One possibility for the receptor site responsible for the binding pattern of hydrophobic cells is the extracellular matrix (ECM) . In this study, we evaluated the interaction of hydrophobic and hydrophilic C . albicans with ECM proteins immobilized onto wells of microtitre tissue . Culture plates, and the ability of ECM proteins to block the binding of hydrophobic cells to splenic tissue ex vivo . Hydrophobic C . albicans bound in greater numbers than hydrophilic cells to the immobilized proteins, particularly fibrinogen, fibronectin, collagen type IV and laminin . Similar results were obtained regardless of C . albicans strains or of growth medium . Collagen and fibronectin blocked the binding of hydrophobic cells to the white pulp but not to the marginal zones in splenic tissues when tested with the ex vivo assay . These results suggests that the diffuse binding pattern of hydrophobic cells in the ex vivo assay may be due to their enhanced ability over hydrophilic cells to bind to ECM proteins, particularly fibronectin and collagen type IV.

J Med Vet Mycol, 1995 Mar-Apr, 33(2), 105 - 11
Cloning of cDNAs coding for Candida albicans cell surface proteins; Sentandreu M et al.; Two cDNA libraries were constructed from mRNAs obtained from yeast cells and germ-tubes of Candida albicans in lambda gt11 . Immunoscreening with polyclonal antibodies raised against cell wall components allowed the detection of 29 positive clones . Two of these clones were selected for their specific reactivity with antisera either from yeast (clone 11Y) or germ-tubes (clone 24M) . cDNA fragments were isolated by the digestion of lambda DNA with EcoRI . Southern blot analysis with these fragments as probes demonstrated homology with C . albicans DNA, and by Northern analysis two mRNAs transcripts were detected with sizes of approximately 1.5 kb for 11Y and 1.1 kb for 24M . Both transcripts were present in yeast cells as well as in germ-tubes . The whole genes were isolated from a C . albicans genomic library in the YRp7 vector by hybridization with the cDNA probes . Monospecific antibodies were purified from polyclonal antisera by affinity for the fusion proteins . Western blot analysis with 11Y-specific antibodies revealed a cross-reactivity with material found in the yeast cell wall as well as in other subcellular fractions, whereas clone 24M codes for a 30 kDa protein detected mainly in the membrane fraction and in the SDS-solubilized material from mycelial cell walls . Sequencing of the cDNA molecules and restriction map of the cloned genes demonstrate that clone 11Y is an enolase previously characterized in C . albicans, whereas clone 24M does not show significant homology with any other cloned gene.

Mol Biochem Parasitol, 1995 Mar, 70(1-2), 119 - 29
Purification and characterization of a soluble nucleoside diphosphate kinase in Trypanosoma cruzi; Ulloa RM et al.; A soluble nucleoside diphosphate kinase (NDP kinase) was purified and characterized in epimastigote forms of Trypanosoma cruzi . The enzyme was purified by affinity chromatography on Blue-agarose and Q-Sepharose columns and by FPLC on a Superose 12 column . A membrane-associated NDP kinase was identified which accounts for 30% of total enzymatic activity . Western blot analysis of the soluble NDP kinase revealed a 16.5-kDa monomer recognized by polyclonal antibodies to NDP kinase from Dictyostelium discoideum, Candida albicans or human . Most of the T . cruzi NDP kinase is found in the cell as a hexamer composed of 16.5-kDa monomers . The Km values of the enzyme for ATP, GDP and dTDP were 0.2 +/- 0.008 mM, 0.125 +/- 0.012 mM and 0.4 +/- 0.009 mM, respectively . The parasite enzyme was stable, remained active at 65 degrees C and was found to tolerate up to 2.5 M urea . The 16.5-kDa subunit was phosphorylated with {gamma-32P}ATP or thiophosphorylated with {35S}GTP gamma S . The incubation of the 32P-labelled phosphoenzyme with unlabelled nucleoside 5'-diphosphates resulted in the formation of 32P-labelled nucleoside 5'-triphosphates without strict base specificity, indicating that the reaction mechanism of the T . cruzi enzyme is the same as reported for other NDP kinases . When the phosphoenzyme was incubated with a mixture of nucleoside 5'-diphosphates, GTP was preferentially formed.

Bratisl Lek Listy, 1995 Mar, 96(3), 141 - 3
{Treatment of mycotic skin infections}; Simaljakova M; Mycotic infections of the skin remain still to represent a therapeutic problem . The development of new antimycotics widens the possibilities of therapy . One of them is oxiconasolnitrate which is a derivate of of imadazole . It is available on our market as Myfungar cream . The clinical postregistration study verified the effectivity of this preperation . The examined group was constituted of 30 patients . 24 patients were afflicted with epidermophytia inguinalis, 1 patient with epidermophytia cruris, 1 patient with epidermophytia manuum, 1 patient with trichophytia corporis and 2 with candidosis submammaris . Cultivation examination revealed 20 time Trichophyton rubrum, one time Trichophyton mentagrophytes var . interdigitale, one time Trichophyton mentagrophytes var . granulosum, one time Epidermophyton floccosum, and 2 time Candida albicans . Therapy lasted 21-42 days . All patients were treated for a period of 6 weeks . 24 treated cases after the therapy included 10 wit persistent squamming, 4 cases with erythema, 3 cases with infiltration, and 1 case with persistent vesiculation . 4 patients suffered from itch irritation, and 1 from pain . 4 patients were not evaluated for their therapy was interrupted . Cultivation examinations in 5 cases revealed the presence of T . rubrum . Microscopic examinations were positive in 10 patients . Following 6 weeks of therapy 20% of patients remained to yield positive cultivation findings and 40% of patients were microscopically positive . 2 patients with candidosis were clinically cured and their laboratory findings were negative in 5-6 weeks . Myfungar creme is a preparation which had in 80% of patients a ver y good or good therapeutic effect . Its low sensibilization abilities and application once per day are advantageous . (tab . 6, Ref . 6.)

Bratisl Lek Listy, 1995 Mar, 96(3), 122 - 6
{Mycotic infections in childhood}; Simaljakova M et al.; The occurrence of mycotic infections has a rising tendency . Epidemiologic studies investigate this problem in adults . The problem is still insufficiently analyzed in children . The study is aimed at gaining information about the occurrence of mycoses evoked by dermatophytes and yeasts in children living in Slovakia, namely on the basis of the analysis of results for the mycological laboratory . From 1992 to 1993 6,511 samples which included 556 samples from children were processed on the basis of suspective dermatophytoses, the children being at the age of 1 to 15 years . 6,896 samples suspective of yeast infections included 3,620 mycologically positive samples . 279 samples were taken from 265 children at the age of 3 months to 15 years . 556 children with dermatophytosis included 116 mycologically positive cases, 64 boys and 52 girls . Microscopic examination was positive in 90 children and cultivation examination was positive in 85 children . Trichophytosis was confirmed in 38 children (32.8%) microsporiasis in 9 (7.8%), epidermophytosis in 50 (43.1%) and onychomycosis in 19 children (16.3%) . Isolated antropophilic dermatophytes were represented most frequently by Trichophyton rubrum, 26 times, isolated zoophylic species included T . verrucosum, 15 times and T . mentagrophytes var . granulosum 13 times . 29 children (25%) had epidermophytosis and onychomycosis of feet . The group of 265 children with yeast infections included 123 with primary skin disease and 134 with internal disease . The most frequently monitored materials were stools, 147 times, and scrapings of the oral cavity, 78 times . The total number of isolated species of yeasts was 17 . Candida albicans was isolated 218 times (78.2%) . The total number of examinations included 7.1-8.5% of samples from children.(ABSTRACT TRUNCATED AT 250 WORDS)

Curr Genet, 1995 Mar, 27(4), 320 - 9
Molecular cloning and characterization of a novel gene of Candida albicans, CDR1, conferring multiple resistance to drugs and antifungals; Prasad R et al.; By functional complementation of a PDR5 null mutant of Saccharomyces cerevisiae, we have cloned and sequenced the multidrug-resistance gene CDR1 of Candida albicans . Transformation by CDR1 of a PDR5-disrupted host hypersensitive to cycloheximide and chloramphenicol resulted in resistance to cycloheximide, chloramphenicol and other drugs, such as the antifungal miconazole, with collateral hypersensitivity to oligomycin, nystatin and 2,4 dinitrophenol . Our results also demonstrate the presence of several PDR5 complementing genes in C . albicans, displaying multidrug-resistance patterns different from PDR5 and CDR1 . The nucleotide sequence of CDR1 revealed that, like PDR5, it encodes a putative membrane pump belonging to the ABC (ATP-binding cassette) superfamily . CDR1 encodes a 1501-residue protein of 169.9 kDa whose predicted structural organization is characterized by two homologous halves, each comprising a hydrophobic region with a set of six transmembrane stretches, preceded by a hydrophilic nucleotide binding fold.

FEMS Immunol Med Microbiol, 1995 Mar, 11(1), 69 - 72
Cleavage of human big endothelin-1 by Candida albicans aspartic proteinase; Tsushima H et al.; A Candida albicans aspartic proteinase (CAP), one of the secretory proteinases of Candida albicans, is thought to be a possible virulence factor in Candida albicans infection . Whereas endothelin-1 is found as an endothelium-derived strong vasoconstrictive peptide, it is known to have a role in the maintenance of vascular homeostasis and tissue survival . Endothelin-1 is generated from a precursor form of endothelin-1, the so-called big endothelin-1 . It has recently been reported that cathepsin D, E and pepsin, which are aspartic proteinases, convert big endothelin-1 to endothelin-1 . In this study, the relationship between CAP and big endothelin-1 was studied . High performance liquid chromatography analysis revealed that big endothelin-1 was cleaved into several amino acid sites by CAP, but endothelin-1 was not converted from big endothelin-1 . CAP cleaved big endothelin-1 at different sites when compared with that of other known aspartic proteinases, and it suppressed endothelin-1 production through the degradation of big endothelin-1 . CAP may break homeostatic mechanism of endothelin-1 in Candida albicans infectious lesions.

FEMS Immunol Med Microbiol, 1995 Mar, 11(1), 57 - 63
Adherence of germ tubes of Candida albicans to tissues from immunocompromised mice; Vespa MN et al.; The influence of immune status of the host on binding of germ tubes of Candida albicans to murine tissue sections in an ex vivo assay was examined . Generally, germ tubes appeared randomly adhered to the tissues examined and binding was unaffected by immunodeficiency induced by treatment with cyclophosphamide and cortisone acetate . Adherence was somewhat reduced in spleen and kidney sections or increased in liver sections and unchanged in lymph node sections from treated mice compared to sections from control animals . Scanning electron micrographs showed organisms appeared to be loosely or tightly bound to the surface or partially embedded in spleen sections from both control and treated mice . These observations suggested that qualitative and quantitative difference in adhesion of germ tubes to various tissues may contribute little to the susceptibility of the immunodeficient animal to candidal infection.

FEMS Immunol Med Microbiol, 1995 Mar, 11(1), 45 - 55
Topical application of a corticosteroid destabilizes the host-parasite relationship in an experimental model of the oral carrier state of Candida albicans; Deslauriers N et al.; Using an experimental model in the mouse we have shown that both local and central lines of defense, involving CD4+ T cells, participate in a dynamic interaction to maintain a long-term carrier state of Candida albicans in the oral cavity . We have tested the impact of a predisposing factor to oral candidiasis in the form of a topical application of a corticosteroid (Topsyn gel) to the oral mucosa for 75 mice twice a day for a 20-day period . Very rapidly after the treatment was initiated, i.e . on day 4, the residual population of Candida increased up to 40-fold and by day 21, the population was 400-fold that of the carrier state . The resident population of intraepithelial CD4+ T cells in the oral mucosa virtually disappeared during the treatment . A topical corticosteroid application also resulted in a massive depletion of T cells in the lymph nodes and in the transient abrogation of the DTH reaction to Candida antigens . On cessation of treatment, normal levels of both Candida and intraepithelial CD4+ T cells were also quickly restored . These results suggest that resistance to superficial invasion by Candida is linked to the presence of an oral mucosal line of defense and that topical application of corticosteroids may dramatically shift the host-parasite relationship in favor of Candida.

Acta Otorrinolaringol Esp, 1995 Mar-Apr, 46(2), 85 - 9
{Otomycosis and topical application of thimerosal: study of 152 cases}; Tisner J et al.; PURPOSE: To evaluate the effectiveness of the topical application of Timerosal (merthilate tintura) in mycosis involving the external auditory canal . PATIENTS AND METHODS: The study includes 152 patients with the clinical, otoscopic and microscopic diagnosis of otomycosis . Results were assessed 72 hours and 10 days after the application . RESULTS: Bacteriological study was performed in 83 patients, finding Aspergilly niger in 54.0% of the cases, Candida albicans in 25.4%, Aspergillus fumigatus in 15.8% and Penicillium in 4.8% . Improvement at 72 h . was found in 66.4% and at 10 days in 93.4% of the patients . Bacteriological contamination was found in 6.6% of the total . COMMENTS: In most of the patients, the otomycosis healed after cleaning of the external auditory canal and topical application of timerosal . This method is easy to apply, fast, effective, of low cost and few side effects.

Mycoses, 1995 Mar-Apr, 38(3-4), 119 - 23
Mycoserological study of the treatment of paediatric cystic fibrosis patients with Saccharomyces boulardii (Saccharomyces cerevisiae Hansen CBS 5926); Muller J et al.; Saccharomyces boulardii (SB) (Saccharomces cerevisiae Hansen CBS 5926) is a yeast widely used in humans for the prevention and treatment of infectious enterocolitis . SB is said also to antagonize Candida albicans when given orally to living organisms . This double-blind trial was performed to determine the effect and tolerance of SB as an oral therapeutic in patients suffering from cystic fibrosis receiving long-term treatment with cephalosporins or cotrimoxazole, by examining C . albicans counts in the intestine . Extensive mycoserological examinations for drug safety evaluation were also performed . To be selected for the study patients had to present C . albicans in their intestinal flora . None of the patients enrolled exhibited clinical symptoms of candidosis . A daily dose of 750 mg (250 mg t.i.d.) of lyophilized SB given for 21 days did not affect the number of C . albicans commensals in those patients . However, the mycoserological data confirmed the safety of SB treatment with respect to a hypothetically possible SB fungaemia and a possible falsification of Candida serology.

Mycoses, 1995 Mar-Apr, 38(3-4), 107 - 10
Identification of oral yeast species isolated from individuals with diabetes mellitus; Aly FZ et al.; In our epidemiological study of 439 patients with diabetes mellitus, the proportion of Candida albicans isolated by the oral rinse technique was 67% . A comparison of the conventional germ tube test with the API 20C Auxanogram kit revealed that 23.6% (129/546) of germ tube-positive species were not identified as C . albicans by the kit . The API 20C Auxanogram therefore underestimated the prevalence of C . albicans . Additionally, a significant number of yeasts (138/1050, 13.1%) isolated from these patients could not be reliably identified by the kit.

FEMS Microbiol Lett, 1995 Feb 15, 126(2), 177 - 80
Avirulence of Candida albicans auxotrophic mutants in a rat model of oropharyngeal candidiasis; Cole MF et al.; The virulence of Candida albicans strain SC5413 and two isogenic derivatives have been investigated in a rat model of oropharyngeal candidiasis . The results demonstrate that both mutant strains are avirulent in this animal model while the parental strain readily initiates infection . Avirulence is not related to altered growth characteristics or the inability of the strains to undergo yeast-to-hyphal morphogenesis . The potential importance of nutritional sufficiency as a virulence factor as well as the possibility of utilizing such strains in the development of an in vitro expression technology system for Candida albicans is discussed.

Mol Gen Genet, 1995 Feb 6, 246(3), 342 - 52
The frequency of integrative transformation at phase-specific genes of Candida albicans correlates with their transcriptional state; Srikantha T et al.; The phase transition between the white and opaque phenotypes in the switching system of Candida albicans strain WO-1 is accompanied by the differential expression of the white-specific gene WH11 and the opaque-specific gene PEP1 . The frequency of integrative transformation at the white-specific gene locus WH11 is between 4.5 and 7.0 times more frequent in white than in opaque spheroplasts, and the frequency of disruptive transformation at the opaque-specific gene locus PEP1 is 30.5 times more frequent in opaque spheroplasts than in white spheroplasts . In contrast, the frequencies of integrative transformation at the constitutively expressed loci ADE2 and EF1 alpha 2 are similar in the white and opaque phases . Therefore, the frequency of integration of linear plasmid DNA containing sequences of phase-specific genes correlates with the transcriptional state of the targeted locus.

Ginekol Pol, 1995 Feb, 66(2), 117 - 20
{Prophylaxis of puerperal infections using the Gyno-Pevaryl 150 preparation}; Poludniewski G et al.; The authors estimated the biocenosis of vagina in women before delivery . The examination were made upon 50 women between 28 and 40 week of pregnancy and on the third day after delivery; comparing the results of estimation the purity of vagina and bacterial inoculations in two groups of pregnant women:-control group (without G-P 150 treatment) and in group with G-P 150 application . The authors proved the entire disappearance of mycotic infection (candida albicans) in women treated with G-P 150 . The also stated that potential pathogenic bacterial flora during puerperium occurs much more rarely.

FEMS Microbiol Lett, 1995 Feb 1, 126(1), 93 - 6
A purine permease in Candida glabrata; Sen Gupta S et al.; Competition experiments revealed that adenine and guanine were transported by a purine permease in both Candida glabrata 4 and a C . glabrata 4 cytosine permease negative mutant . The C . glabrata 4 cytosine permease negative mutant was isolated using 5-fluorocytosine selection . This mutant no longer transported cytosine, but transported adenine and guanine . A transport system for hypoxanthine was not detected . Hence, in addition to the cytosine permease, a purine permease exists in C . glabrata . This differs from the purine cytosine permeases in Saccharomyces cerevisiae and Candida albicans which transport adenine, cytosine, guanine and hypoxanthine.

J Am Podiatr Med Assoc, 1995 Feb, 85(2), 104 - 15
Candida albicans, the opportunist . A cellular and molecular perspective; Dupont PF; Candida albicans causes the majority of opportunistic fungal infections . The yeast's commensualistic relationship with humans enables it, when environmental conditions are favorable, to multiply and replace much of the normal flora . Virulence factors of C . albicans, enabling the organism to adhere to and penetrate host tissues, involve specific molecular interactions between the cells of the fungus and the host . Localized disease, such as oral candidiasis, onychomycosis, and vaginitis, results . These infections are usually limited to surfaces of the host, and can be quickly and successfully controlled by the use of one of the available antifungal agents . Candida albicans infections typically become systemic and life threatening when the host is immunocompromised . Depending on the immune defect in the host, one of the spectrum of Candida diseases can develop . If successful treatment of these patients is to be achieved, modulation of the immune deficit, as well as the use of an appropriate antifungal drug, must become a routine part of therapeutic interventions.

Toxicol Appl Pharmacol, 1995 Feb, 130(2), 316 - 21
Biochemical and functional analysis of rat bronchoalveolar macrophages containing chemically induced phospholipid inclusions; Waites CR et al.; Cationic amphiphilic drugs (CADs) are structurally characterized by hydrophobic ring structures and hydrophilic side chains . Studies have demonstrated that repeated administration of CADs to experimental animals and humans may induce phospholipid (PL) accumulation within the cells of various tissues . The immunomodulatory azaspiranes are novel CADs with beneficial effects in a number of animal models of autoimmune disease and transplantation . Although the mechanism of action of these compounds is unclear, efficacy in all of the disease models is accompanied by the generation of suppressor cell (SC) activity in various lymphoid organs . SK&F 105685 (N,N-dimethyl-8,8-dipropyl-2-azaspiro{4,5}decane-2-propanamine+ ++ hydrochloride) and two analogs, SK&F 106615 and SK&F 103811, were compared with chlorphentermine and chloroquine for their ability to induce PL accumulation and SC activity . Oral administration of SK&F 105685 and SK&F 106615 caused PL accumulation in bronchoalveolar lavage macrophages (AM) but to a far lesser extent (three- to fivefold) than chlorphentermine . Neither the immunologically unreactive azaspirane SK&F 103811 nor chloroquine affected PL levels . AM from rats treated with SK&F 105685 or SK&F 106615 expressed more potent SC activity than chlorphentermine . Thus, SC activity did not correlate with the extent of PL accumulation . Neither SK&F 103811 nor chloroquine induced SC activity . AM from SK&F 105685-treated rats had an enhanced ability to kill the opportunistic pathogen Candida albicans in vitro indicating that there was no impairment of macrophage-dependent host defense mechanisms.

Am J Clin Pathol, 1995 Feb, 103(2), 130 - 5
Specific immunohistochemical identification of Candida albicans in paraffin-embedded tissue with a new monoclonal antibody (1B12); Monteagudo C et al.; In invasive candidiasis, the identification of Candida organisms in tissue samples or in normally sterile fluids is essential for an accurate diagnosis . Species identification is an important clue for the source of infection and in epidemiological studies . In this article, the authors have tested the value of a new monoclonal antibody (1B12) to detect C albicans in culture by immunofluorescence, and in tissue samples by immunohistochemistry . MAb 1B12 was found to specifically recognize C albicans, does not cross-react with other Candida species or other structurally similar fungi, and is very sensitive and specific in paraffin-embedded tissue, having no reactivity in normal human tissues or necrotic areas . Therefore, MAb may be a valuable tool in the evaluation of fungal infections in paraffin-embedded tissue, particularly when Candida species identification is needed.

J Leukoc Biol, 1995 Feb, 57(2), 287 - 96
Transforming growth factor beta modulates C3 and factor B biosynthesis and complement receptor 3 expression in cultured human monocytes; Hogasen AK et al.; Complement biosynthesis in monocytes is stimulated by different pathogens and modulated by a variety of cytokines, but little is known about the possible effect of transforming growth factor beta (TGF-beta) on this monocyte function . We therefore studied the effect of TGF-beta 1 and TGF-beta 2 on constitutive, lipopolysaccharide (LPS)- and Candida albicans-induced monocyte biosynthesis of complement components C3 and factor B . Under all three conditions, both forms of TGF-beta (20 ng/ml) induced a two- to fourfold increase in C3 concentration in monocyte supernatants harvested after 2 or 5 days of cell culture, an effect that was abrogated by cycloheximide . In contrast, constitutive and pathogen-induced production of factor B was suppressed by TGF-beta . The effects of TGF-beta on complement production were neutralized by a monoclonal anti-TGF-beta antibody . Moreover, TGF-beta suppressed the pathogen-induced release of granulocyte-macrophage colony-stimulating factor and down-regulated the expression of complement receptor 3 (CD11b/CD18), while the expression of CD11a/CD18, a related beta 2 integrin, was unaffected . These novel effects of TGF-beta emphasize the immunomodulatory significance of this cytokine.

J Infect Dis, 1995 Feb, 171(2), 393 - 9
Pharmacologic inhibitors of tumor necrosis factor production exert differential effects in lethal endotoxemia and in infection with live microorganisms in mice; Netea MG et al.; Tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) are principal mediators of septic shock; inhibition of TNF-alpha production may ameliorate outcome in severe infections . Pentoxifylline, chlorpromazine, and thalidomide inhibit TNF-alpha production . Their effects were tested in lethal endotoxemia in sensitized mice . Only chlorpromazine significantly improved survival . Chlorpromazine and pentoxifylline significantly reduced postendotoxin circulating TNF-alpha, by 89% and 76%, respectively . Chlorpromazine also significantly reduced IL-1 beta and soluble TNF receptor-P75 . No drug improved survival in Klebsiella pneumoniae-infected mice despite significantly lower circulating TNF-alpha concentrations in chlorpromazine- or pentoxifylline-treated animals . The three compounds decreased circulating TNF-alpha in Candida albicans-infected mice, but survival was not influenced . In neutropenic mice, chlorpromazine had no influence on candida in organs, but in normal mice, Candida counts in kidneys were higher in chlorpromazine-treated mice . Thus, inhibition of TNF-alpha production was of no benefit in K . pneumoniae infection and worsened outcome in C . albicans infection.

Mol Cell Biol, 1995 Feb, 15(2), 601 - 13
PHR1, a pH-regulated gene of Candida albicans, is required for morphogenesis; Saporito-Irwin SM et al.; Candida albicans, like many fungi, exhibits morphological plasticity, a property which may be related to its biological capacity as an opportunistic pathogen of humans . Morphogenesis and alterations in cell shape require integration of many cellular functions and occur in response to environmental signals, most notably pH and temperature in the case of C . albicans . In the course of our studies of differential gene expression associated with dimorphism of C . albicans, we have isolated a gene, designated PHR1, which is regulated in response to the pH of the culture medium . PHR1 expression was repressed at pH values below 5.5 and induced at more alkaline pH . The predicted amino acid sequence of the PHR1 protein was 56% identical to that of the Saccharomyces cerevisiae Ggp1/Gas1 protein, a highly glycosylated cell surface protein attached to the membrane via glycosylphosphatidylinositol . A homozygous null mutant of PHR1 was constructed and found to exhibit a pH-conditional morphological defect . At alkaline pH, the mutant, unlike the parental type, was unable to conduct apical growth of either yeast or hyphal growth forms . This morphological aberration was not associated with defective cytoskeletal polarization or secretion . The results suggest that PHR1 defines a novel function required for apical cell growth and morphogenesis.

Infect Immun, 1995 Feb, 63(2), 569 - 72
Adherence of Candida albicans to human buccal epithelial cells: host-induced protein synthesis and signaling events; Bailey A et al.; The synthesis of proteins by Candida albicans was studied following adherence of blastoconidia to human buccal epithelial cells (HBEC) . Initially, labeling of HBEC, C . albicans, and HBEC-C . albicans with {35S}methionine was performed . After a 3-h incubation and prior to labeling with {35S}methionine, the cultures were treated with cycloheximide to prevent HBEC protein synthesis . The HBEC-C . albicans mixture as well as C . albicans and HBEC incubated separately were extracted with beta-mercaptoethanol (beta-ME) . These extracts as well as the cell residue (solubilized by boiling with sodium dodecyl sulfate {SDS}) were examined by SDS-polyacrylamide gel electrophoresis and autoradiography . In comparison to cultures of C . albicans incubated without HBEC, proteins with molecular masses of approximately 52 to 56 kDa from beta-ME extracts and from SDS-solubilized cells were observed only from adhering cultures . In addition, unlabeled beta-ME extracts were electrotransferred to nitrocellulose and immunoblotted with antiphosphotyrosine antibodies to determine whether cell signaling events were occurring during adherence . Proteins with molecular masses of 54 and 60 kDa were recognized only in mixed cultures of C . albicans and HBEC . These data indicate that following adherence of C . albicans to HBEC, new Candida proteins are expressed . Further, these events are accompanied by the expression of signal proteins, presumably of Candida origin.

Infect Immun, 1995 Feb, 63(2), 547 - 53
Mice immunized by primary vaginal Candida albicans infection develop acquired vaginal mucosal immunity; Fidel PL Jr et al.; It has been postulated that systemic cell-mediated immunity (CMI) is an important host defense mechanism against Candida infections of the vagina . However, in an estrogen-dependent murine model of experimental vaginal candidiasis, we recently showed that systemic Candida-specific Th1-type CMI induced by immunization with Candida culture filtrate antigen had no effect on vaginal Candida population levels during the course of a vaginal infection . In the present study, mice given a second vaginal inoculation in the presence of peripheral Candida-specific Th1-type CMI induced by prior vaginal infection had anamnestic-type increased delayed-type hypersensitivity (DTH) responses, concomitant with significantly fewer Candida organisms in the vagina than in primary-infected mice . In addition, organisms in secondary-infected mice were fragmented and superficial penetration into the epithelium was reduced . The systemic presence of Candida-specific T suppressor (Ts) cells that significantly suppressed the infection-derived anamnestic DTH reactivity did not abrogate the protective effect in the vagina . Additional experiments showed that vaginally immunized mice were not protected from gastrointestinal or systemic candidiasis and, in contrast to mice with a second vaginal infection, did not demonstrate anamnestic DTH reactivity . These results suggest that a moderate level of local protection against a Candida vaginal infection can be achieved by vaginal immunization but that the protective role of acquired peripheral Candida-specific Th1-type reactivity at the vaginal mucosa appears to be limited.

Int Arch Allergy Immunol, 1995 Feb, 106(2), 118 - 23
Nitrocellulose-RAST analysis of allergenic cross-reactivity of Candida albicans and Saccharomyces cerevisiae mannans; Nermes M et al.; Two chemically purified mannan preparations and one affinity purified mannan preparation of Candida albicans and Saccharomyces cerevisiae were analyzed for allergenic cross-reactivity . Simultaneous IgE binding in radioallergosorbent test (RAST) was studied with all preparations and the chemically purified mannans were also used for RAST inhibitions . The yeast mannans were purified with the Peat method, using repeated ethanol precipitations and Fehling's precipitation in alkaline conditions, and the Cetavlon method, using hexadecyltrimethylammonium bromide and ethanol precipitations . Affinity purification of the mannans was performed with concanavalin A lectin bound to a Sepharose column . Mannan from Pityrosporum ovale was purified with the Cetavlon method for analysis of simultaneous RAST reactivity . Simultaneous IgE binding to all the studied yeast mannans was generally seen in all the studied sera of patients suffering from atopic dermatitis . The strongest responses were seen against C . albicans mannan, especially the Cetavlon mannan . In the RAST inhibition studies IgE binding to S . cerevisiae mannan was completely inhibited by C . albicans mannan preparations, whereas reciprocal inhibition was not complete . These results indicate that in atopic dermatitis simultaneous IgE response to yeast polysaccharides occurs and that the major sensitizer is C . albicans and IgE antibodies against S . cerevisiae mannan are cross-reacting.

Acta Paediatr, 1995 Feb, 84(2), 183 - 7
Prospective evaluation of Candida antigen and antibody assays for detection of Candida infections in children with malignant disease; Ormala T et al.; The clinical efficacy of assays for Candida albicans antigens by latex agglutination and for antibodies by indirect haemagglutination were prospectively evaluated in the diagnosis of invasive Candida infections in 38 children suffering from acute leukaemia or other malignant disease . The controls were 74 other patients without any malignancy; 72 of these had no signs or symptoms of fungal infections, but 2 had an invasive C . albicans infection . During a period of 21 months, 302 serum samples were tested by both assays, and the results were compared with clinical and other microbiological data . Invasive fungal infection was diagnosed on clinical grounds in 2 of the immunocompromised children, and periodic gut colonization was demonstrated in 11 of 36 (31%) children in this group . Positive Candida antigen was detected in 14 patients (37%) and a positive antibody titre in 7 patients (18%) . Colonization was not correlated with antigen or antibody titre . Compared with the presence of invasive fungal infection, the antibody assay detected all four infections, whereas the antigen assay detected one of the two C . albicans septicaemias . Although the Candida antibody assay performed well, a detectable change in antibody titres appeared only slowly . Thus it was of no clinical help when antifungal treatment was to be considered . Follow-up of antibody titres, however, gave confirmation of the presence of fungal infection as well as the response to antifungal treatment.

Neurosurgery, 1995 Feb, 36(2), 411 - 2
Magnetic resonance image findings of spinal intramedullary abscess caused by Candida albicans: case report; Lindner A et al.; We present the clinical, serological, and radiological features of a patient with a spinal intramedullary abscess caused by Candida albicans . Antimycotic treatment was successful, and no neurosurgical approach was necessary.

Antimicrob Agents Chemother, 1995 Feb, 39(2), 422 - 6
Multidrug resistance in Candida albicans: disruption of the BENr gene; Goldway M et al.; The BENr gene of Candida albicans, which confers resistance on susceptible strains of Saccharomyces cerevisiae to six structurally and functionally unrelated drugs, was described recently (R . Ben-Yaacov, S . Knoller, G . Caldwell, J . M . Becker, and Y . Koltin, Antimicrob . Agents Chemother . 38:648-652, 1994) . This gene bears similarity to membrane proteins encoding antibiotic resistance in prokaryotes and eukaryotes . The effect of disruption of this gene on viability and drug susceptibility was determined . The results indicate that the gene is not essential but its inactivation leads to susceptibility to three of the four drugs tested . Inactivation of this gene did not increase the susceptibility of the mutant to benomyl, suggesting that C . albicans has other mechanisms of resistance, some of which may be additional efflux pumps that confer resistance to this tubulin-destabilizing agent.

Can J Microbiol, 1995 Feb, 41(2), 136 - 44
Antimicrobial mode of action of secretions from the metapleural gland of Myrmecia gulosa (Australian bull ant); Mackintosh JA et al.; Secretions from exocrine metapleural glands of Myrmecia gulosa (Australian bull ant) exhibit broad-spectrum antimicrobial activity . Treatment of the yeast Candida albicans with metapleural secretion resulted in the rapid and total leakage of K+ ions from cells within 10 min . Ultrastructural analysis of the bacteria Bacillus cereus, Escherichia coli, and Pseudomonas aeruginosa, and cells and protoplasts of Candida albicans demonstrated gross damage of the cell membrane and aggregation of the cytoplasmic matrix of treated cells . Degradation of membrane-bound organelles was also observed in Candida albicans . The antimicrobially active components of metapleural secretions were nonpolar and interacted with the phospholipid bilayer, causing damage to the structural integrity of liposomes and the release of carboxyfluorescein . The data suggest that the antimicrobial agents in metapleural secretion act primarily by disrupting the structure and function of the phospholipid bilayer of the cytoplasmic membrane.

J Clin Microbiol, 1995 Feb, 33(2), 432 - 9
Reduced inhibition of Candida albicans adhesion by saliva from patients receiving oral cancer therapy; Umazume M et al.; The effect of saliva on the adhesion of Candida albicans to epithelial cells was examined in vitro by using saliva from healthy controls and patients with oral squamous cell carcinoma . The adhesion of C . albicans to established epithelial tumor cells was reduced by 40% by salivary treatment of the C . albicans or epithelial cells . The inhibitory activity of saliva was almost completely abolished by anti-secretory immunoglobulin A antibody, concanavalin A, and mannose . Compared with saliva from healthy individuals, that from patients who had received chemoradiotherapy for oral carcinoma showed reduced suppression of C . albicans adhesion, which accompanied decreased salivary secretory immunoglobulin A and lactoferrin concentrations . A greater number of C . albicans cells adhered to buccal cells obtained from patients who had received chemoradiotherapy than to those from healthy individuals . Treatment of either epithelial cells or C . albicans with anticancer drugs induced an increase in adherence of epithelial cells and yeast cells . In contrast, concanavalin A- and mannose-pretreated C . albicans exhibited reduced adhesion to epithelial cells . No further decrease of C . albicans adhesion was observed when both epithelial cells and yeast phase C . albicans were treated with mannose . In conclusion, the inhibition of C . albicans adhesion by saliva depends largely on mannose residues on salivary glycoproteins and mannose is one of the binding ligands on both C . albicans and epithelial cells . In addition, anticancer therapy may induce oral C . albicans overgrowth by decreasing salivation and the concentrations of glycoproteins in saliva inhibiting C . albicans adhesion and by increasing the adhesive properties of both C . albicans and oral epithelial cells.

J Antibiot (Tokyo), 1995 Feb, 48(2), 126 - 33
Helioferins; novel antifungal lipopeptides from Mycogone rosea: screening, isolation, structures and biological properties; Grafe U et al.; Helioferins A and B were detected as novel aminolipopeptides in cultures of Mycogone rosea DSM 8822 in the course of a screening for mediators of helianthate anion transfer from aqueous to toluene phases . Their structures as novel antibiotics and cytotoxic agents were elucidated by mass spectrometry and NMR spectroscopic methods . Antimicrobial activity was estimated against Candida albicans and Gram-positive bacteria including Mycobacterium spp.

Microbiology, 1995 Feb, 141 ( Pt 2), 469 - 76
Analysis of the chromosomal localization of the repetitive sequences (RPSs) in Candida albicans; Chindamporn A et al.; The location and organization of repetitive sequences, members of the RPS family, which are sequences specific to Candida albicans, were determined on each chromosome of C . albicans strain FC18 . Using pulsed-field gel electrophoresis, we separated seven fractions from eight chromosomes . Each chromosome was cleaved by BamHI and XhoI to excise the RPSs, which were then detected by hybridization with an RPS probe . All chromosomes except chromosome 4 carried RPSs, and these RPSs were located within a limited region on each chromosome . From the digestion of each chromosome with SfiI and probing with the RPSs, we found that these recognition sites within the RPS region were conserved among all RPS-containing chromosomes . For further characterization of the RPSs, the locations and the boundary regions of the RPSs were examined on chromosome 6 of strain FC18 as a model chromosome . Using the restriction enzymes SfiI, SmaI, XhoI, BamHI, MluI and NruI, we constructed a semi-macro physical map of the RPSs and their boundary regions on this chromosome . We also determined which part of the RPS was adjacent to each boundary by using sub-fragments of RPS as probes . The physical configuration around the RPSs and their boundary regions are presented . The results obtained should be useful for future analysis of the function of these regions.

Comp Immunol Microbiol Infect Dis, 1995 Feb, 18(2), 105 - 13
Seasonal variations in the immune system of the cyprinid Tinca tinca . Phagocytic function; Collazos ME et al.; Seasonal variations in the in vitro phagocytic process of blood granulocytes from the tench Tinca tinca were examined . Different stages of the phagocytic process: mobility rate, attachment, ingestion and killing of Candida albicans were evaluated . Tench were kept in natural ponds in ambient water temperature, and the in vitro assays were performed at both 22 degrees C and the relative ambient temperature . Results between the seasonal samples were then compared . In vitro induced mobility, attachment, ingestion and killing of C . albicans showed strong seasonal variations, furthermore, the phagocytic process at 22 degrees C varied significantly according to season . Phagocytic activity from samples taken during the spring demonstrated the highest activity at 22 degrees C, whilst greatest activity at seasonal temperature, in terms of mobility rate, phagocytic index and microbicide capacity, occurred during the winter . These results are consistent with the hypothesis that phagocytosis in fish is resistant to low temperatures.

J Nutr Sci Vitaminol (Tokyo), 1995 Feb, 41(1), 127 - 37
Alteration of the respiratory burst and phagocytosis of macrophages under protein malnutrition; Teshima S et al.; To understand the role of macrophages in impaired host defense under protein malnutrition (PM), we examined the activities of the respiratory burst and phagocytosis of resident peritoneal macrophages from weaning female mice fed 5% casein or 5% soy protein isolate (SPI) diet for 14 days . Resident macrophages from the low-protein diet groups released larger amounts of superoxide anion (O2-) after stimulation by phorbol 12-myristate 13-acetate (PMA) or opsonized zymosan than those from the 20% casein and 20% SPI diet groups . Activation of macrophages from protein-deficient mice in vitro with lipopolysaccharide (LPS) or macrophage colony-stimulating factor (M-CSF) under LPS-free conditions did not further enhance O2- production . In spite of the increased O2- production with opsonized zymosan, macrophages from protein-deficient mice did not show any acceleration of phagocytosis of Candida albicans in the presence of normal serum . Our results confirm that the phagocytic function of macrophages is susceptible to PM, and suggest that functional alterations of macrophages may be involved in the failure of development of a specific immune response under PM . Furthermore, the enhanced production of oxygen intermediates by macrophages may augment tissue damage under PM.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1995 Feb, 79(2), 170 - 4
Fluconazole response patterns in HIV-infected patients with oropharyngeal candidiasis; Diz Dios PD et al.; A prospective study to assess the efficacy of fluconazole in oropharyngeal candidiasis in patients with HIV was conducted . A cohort of 30 HIV-positive persons with clinical and microbiologic confirmed oropharyngeal candidiasis (Candida albicans > 1000 CFU/ml) received fluconazole 100 mg daily for 7 days . In vitro antifungal susceptibility tests demonstrated a lack of fluconazole resistances . Cultures of mouth swabs were performed at the end of therapy and 2 weeks later . There was a clinical and microbiologic cure in 26 patients (87%) . In 10 of these 26, cultures remained negative after 2 weeks; most of them had CD4 lymphocyte count > 400/ml . In the other 16 patients (53%), cultures showed a microbiologic relapse 2 weeks after treatment . In spite of clinical improvement, treatment failure was observed in four patients, all of them with CD4 lymphocyte count < 50 ml.

J Clin Invest, 1995 Feb, 95(2), 913 - 8
Milk-induced eczema is associated with the expansion of T cells expressing cutaneous lymphocyte antigen; Abernathy-Carver KJ et al.; The extravasation of T cells at sites of inflammation is critically dependent on the activity of homing receptors (HR) involved in endothelial cell recognition and binding . Two such HR (the cutaneous lymphocyte antigen {CLA} and L-selectin) have been shown to be selectively involved in T cell migration to skin and peripheral lymph nodes, respectively . This study was designed to assess the relationship between the organ specificity of an allergic reaction to food and the expression of HR on T cells activated in vitro by the relevant food allergen . Peripheral blood mononuclear cells were isolated from seven milk allergic children with a history of eczema when exposed to milk . All patients had a positive prick skin test and double-blind placebo-controlled food challenge to milk . 10 children with either allergic eosinophilic gastroenteritis or milk-induced enterocolitis and 8 nonatopic adults served as controls . Five-parameter flow cytometry using monoclonal antibodies was used for detection of the specific HR on freshly isolated T cells versus T cell blasts induced by a 6-d incubation with casein, as compared with Candida albicans . After in vitro stimulation with casein, but not C . albicans, patients with milk allergy and atopic dermatitis had a significantly greater percentage of CLA+ T cells (P < 0.01) than controls with milk-induced enterocolitis, allergic eosinophilic gastroenteritis, or nonatopic healthy controls . In contrast, the percentage of L-selectin-expressing T cells did not differ significantly between these groups . These data suggest that after casein stimulation allergic patients with milk-induced skin disease have an expanded population of CLA+ T cells, as compared with nonatopics or allergic patients without skin involvement . We postulate that heterogeneity in the regulation of HR expression on antigen-specific T cells may play a role in determining sites of involvement in tissue-directed allergic responses.

J Biol Chem, 1995 Jan 20, 270(3), 1113 - 22
Existence of branched side chains in the cell wall mannan of pathogenic yeast, Candida albicans . Structure-antigenicity relationship between the cell wall mannans of Candida albicans and Candida parapsilosis; Shibata N et al.; Isolation of side chain oligosaccharides from mannans of Candida albicans NIH B-792 (serotype B) and Candida parapsilosis IFO 1396 strains has been conducted by acetolysis under mild conditions . Structural study of these oligosaccharides by 1H and 13C NMR and methylation analyses indicated the presence of novel branched side chains with the following structures in C . albicans mannan . {sequence: see text} It was observed that the H-1 proton chemical shifts of the second and the third mannose units from the reducing terminus in each oligosaccharide are shifted upfield by substitution with an alpha-linked mannose unit at position 6 of the 3-O-substituted mannose unit . An agglutination inhibition assay between factor 4 serum and cells of Candida stellatoidea IFO 1397 lacking the beta-1,2-linked mannose unit, with oligosaccharides obtained from these mannans, indicated that only the branched oligosaccharides were active . This finding suggests that the branched oligosaccharides correspond to the epitope of antigenic factor 4 . The presence of the branched structure in other mannans was detected by the characteristic H-1-H-2-correlated cross-peak of the alpha-1,2-linked mannose unit connected with the 3,6-di-O-substituted one by two-dimensional homonuclear Hartmann-Hahn spectroscopy.

FEMS Microbiol Lett, 1995 Jan 15, 125(2-3), 143 - 7
Cell wall anchoring to cytoplasmic membrane of Candida albicans; Hazen KC et al.; Cell wall ultrastructure of the opportunistic pathogenic yeast Candida albicans was investigated by stereoscopic freeze-etching technique . Three wall layers were distinguishable by this technique . No clear periplasmic space was evident . Bilayer membrane invaginations were extensive . The outermost regions of the membrane invaginations were lined with thin, spine-like fibrils, which extended into the cell wall . We suggest that the fibrils along the invaginations are involved in anchoring the cell wall to the membrane.

FEMS Microbiol Lett, 1995 Jan 15, 125(2-3), 121 - 6
Molecular biology of yeast exoglucanases; Larriba G et al.; Three exoglucanase (Exg) genes have been reported in Saccharomyces cerevisiae . Gene EXG1 encodes the major isoenzyme (ExgI) . Differential glycosylation of the primary translation product throughout the secretory pathway results in the secretion of several glycoforms . The major glycoform (ExgIb) contains two short carboxypeptidase Y-like oligosaccharides attached to both potential glycosylation sites present in the molecule . A minor glycoform (ExgIa) arises from the former by elongation of the second oligosaccharide . The protein portion is processed in the secretory pathway by the Kex2 protease . Gene EXG2 encodes a 63 kDa polypeptide with 12 potential glycosylation sites . The predicted protein, ExgII, carries a signal peptide at the amino terminus and a glycosyl-phosphatidyl inositol anchoring motif at the carboxyl end . The latter appears responsible for the particulate nature of this isoenzyme, since its elimination results in the secretion of this activity into the culture medium . Gene SSG1 encodes a 52 kDa polypeptide which is specifically synthesized during sporulation of diploids . SSG1 expression is under control of both sexual (a1-alpha 2 element) and nutritional control . Although homozygous ssg1/ssg1 diploid strains are still able to complete sporulation, they exhibited a delay in the appearance of mature asci . Single or double disruption of EXG1 and EXG2 did not result in any relevant phenotype and the triple mutant behaved as ssg1/ssg1 . A ExgI-related enzyme is secreted by Candida albicans . All these four enzymes share 8 highly conserved regions in the same relative positions, indicating that they derived from a common ancestor . However, no clear function has so far been demonstrated for them.

Eur J Biochem, 1995 Jan 15, 227(1-2), 372 - 8
Kinetics of beta-1,3 glucan interaction at the donor and acceptor sites of the fungal glucosyltransferase encoded by the BGL2 gene; Goldman RC et al.; Formation of branched glucan, glucan-glucan cross links, and glucan-chitin cross links most likely involves the action of fungal wall glucanases and transglycosylases . We developed an HPLC assay using radiolabeled substrates in order to study the kinetics of interaction of donor and acceptor molecules with a glucosyltransferase present in the cell walls of both Saccharomyces cerevisiae and Candida albicans . Purified transferase first forms an activated intermediate from a donor beta-1,3 glucan, releasing free disaccharide . The activated intermediate is transferred, in the presence of an appropriate acceptor beta-1,3 glucan, yielding a linear glucan containing a beta-1,6 linkage at the transfer site {Yu, L., Goldman, R., Sullivan, P., Walker, G . & Fesik, S . W . (1993) J . Biomol . NMR 3, 429-441} . An apparent Km of 0.41 mM for the acceptor site was determined using laminaritetraose as the acceptor . An apparent Km of 31 mM for the donor site was determined using increasing concentrations of laminaripentaose, and monitoring formation of laminaribiose . The enzyme functioned as a glucanase at low concentrations of acceptor molecules, with excess H2O competing for reaction at the activated donor site, thus resulting in hydrolysis . However, as the concentration of acceptor increased, the reaction shifted from hydrolysis to glucosyltransfer . The reaction appeared specific for beta-1,3 glucan as acceptor, in as much as no transfer was detected when either hexa-N-acetyl-chitohexaose or maltooligosaccharides were used as acceptors . The roles of such an enzymic activity in cell wall metabolism is discussed in terms of repair, cross linking and incorporation of newly synthesized chains of beta-1,3 glucan into the previously existing cell wall structure.

Experientia, 1995 Jan 15, 51(1), 35 - 9
Biological activity of secondary metabolites from Bupleurum salicifolium (Umbelliferae); Gonzalez JA et al.; Secondary metabolites from Bupleurum salicifolium were tested against viruses, Gram-positive and Gram-negative bacteria, the yeast Candida albicans, the nematodes Globodera pallida and G . rostochiensis, the insect Spodoptera littoralis and the crustacean Artemia salina . These compounds were also tested against tumoral and non-tumoral cell lines . The polyacetylene 8S-heptadeca-2(Z)-9(Z)-diene-4,6-diyne-1,8-diol exhibited toxicity for A . salina and specific antibiotic activity against Gram-positive bacteria . Nine of the lignans and one coumarin showed toxicity for A . salina, and the lignans bursehernin and matairesinol inhibited the hatching of the two nematode species . These are the first lignans that have been reported as affecting phytoparasitic nematodes, and the first natural products known to have an effect on the hatching of G . pallida . Lignans may play a role in the defence mechanisms of potato plants, as allelopathic substances acting against cyst-forming nematodes.

Transplantation, 1995 Jan 15, 59(1), 45 - 50
Liposomal amphotericin B prevents invasive fungal infections in liver transplant recipients . A randomized, placebo-controlled study; Tollemar J et al.; Eighty-six consecutive liver transplant recipients were prospectively randomized in a double-blind, placebo-controlled antifungal prophylaxis study . Seventy-seven patients received 5 days of prophylaxis starting during the transplantation with either liposomal amphotericin B (AmBisome) 1 mg/kg/day or placebo . Among 40 AmBisome-treated patients, no invasive Candida infection was seen during the first month, compared with 5 invasive Candida albicans infections among 37 control patients (P < 0.05) . Furthermore, 1 placebo patient experienced Aspergillus niger pneumonia . Thus, the overall incidence of invasive fungal infections was 0/40 (0%) in the AmBisome group versus 6/37 (16%) in the placebo group (P < 0.01) . Patient survival at 30 days was 92% versus 94% for AmBisome- and placebo-treated patients, respectively . One patient experienced backache related to AmBisome infusion . Two patients had transient thrombocytopenia possibly caused by AmBisome treatment . AmBisome was otherwise well tolerated . The total cost for all antifungal drugs used in both groups was equal . However, prophylaxis with AmBisome was $5000 less expensive than treatment of proven invasive fungal infections among placebo patients.

Regul Pept, 1995 Jan 5, 55(1), 47 - 56
Inhibition of murine peritoneal macrophage functions by sulfated cholecystokinin octapeptide; De la Fuente M et al.; The effect in vitro of the sulfated octapeptide form of cholecystokinin, CCK-8, at concentrations from 10(-12) M to 10(-6) M on several functions of resting peritoneal macrophages from BALB/c mice: adherence to substrate, mobility (spontaneous and directed by chemical gradient or chemotaxis), ingestion of inert particles (latex beads) or cells (Candida albicans), and production of superoxide anion measured by nitroblue tetrazolium reduction was studied . CCK-8, at concentrations from 10(-10) M to 10(-8) M, inhibited significantly all functions studied with the exception of adherence to substrate, which was increased . A dose-response relationship was observed, with a maximum inhibition of macrophage functions found at 10(-8) M . This neuropeptide induced in murine macrophages a significant, but transient, increase of cAMP levels at 60 sec . On the contrary, CCK-8 produced a slight but significant decrease of protein kinase C (PKC) activity at 5 min of incubation . These results suggest that CCK-8 is a negative modulator of several macrophage functions, and that the inhibition of these activities is carried out through an increase of intracellular cAMP levels and a decrease in PKC activity.

Acta Biochim Pol, 1995, 42(4), 497 - 504
Multiple drug resistance in Candida albicans; Prasad R et al.; By functional complementation of a PDR 5 (pleiotropic drug resistance) null mutant of S . cerevisiae, we have recently cloned and sequenced a multidrug resistance gene CDR 1 (Candida Drug Resistance) . Transformation by CDR 1 of a PDR 5 disrupted host hypersensitive to cycloheximide and chloramphenicol resulted in resistance to these as well as other unrelated drugs . The nucleotide sequence of CDR 1 revealed that, like PDR 5, it encodes a putative membrane pump belonging to the ABC superfamily . CDR 1 encodes a protein of 169.9 kDa whose predicted structural organisation is characterised by two homologous halves, each comprising a hydrophobic region, with a set of six transmembrane stretches, preceded by a hydrophilic binding fold . We now have evidence to suggest that there are several PDR homologues present in C . albicans which display multidrug resistance and a collateral sensitivity pattern different from PDR 5 and CDR 1 . The functions of such genes and their products in the overall physiology of C . albicans is not yet established.

Acta Biochim Pol, 1995, 42(4), 481 - 96
Targeting the fungal plasma membrane proton pump; Monk BC et al.; The need for new mechanistic classes of broad spectrum antifungal agents has prompted development of the membrane sector and ectodomain of the plasma membrane proton pumping ATPase as an antifungal target . The fungal proton pump is a highly abundant, essential enzyme in Saccharomyces cerevisiae . It belongs to the family of P-type ATPases, a class of enzymes that includes the Na+,K(+)-ATPase and the gastric H+,K(+)-ATPase . These enzymes are cell surface therapeutic targets for the cardiac glycosides and several anti-ulcer drugs, respectively . The effects of acid-activated omeprazole show that extensive inhibition of the S . cerevisiae ATPase is fungicidal . Fungal proton pumps possess elements within their transmembrane loops that distinguish them from other P-type ATPases . These loops, such as the conformationally sensitive transmembrane loop 1+2, can attenuate the activity of the enzyme . Expression in S . cerevisiae of fully functional chimeric ATPases that contain a foreign target comprising transmembrane loops 1+2 and/or 3+4 from the fungal pathogen Candida albicans suggests that these loops operate as a domain . The chimera containing C . albicans transmembrane loops 1+2 and 3+4 provides a prototype for mutational analysis of the target region and the screening of inhibitors directed against opportunistic fungal pathogens . Panels of mutants with modified ATPase regulation or with altered cell surface cysteine residues are also described . Information about the ATPase membrane sector and ectodomain has been integrated into a model of this region.

Med Dosw Mikrobiol, 1995, 47(3-4), 203 - 212
{Usefulness of the microcolorimetric method of XTT reduction for evaluation of intracellular killing of pathogens}; Burow A et al.; The aim of this study was to evaluate a XTT assay in testing the killing activity of mouse phagocytes in vitro . Live microorganisms converted XTT to water soluble orange formazan in the presence of CQ . Absorption of formazan measured at 492 nm was directly related to the number of viable cells . The percentage of staphylococci killed by granulocytes and macrophages was 10-40% (1 h- and 2 h-respectively), in the cultures containing 10 bacteria/phagocyte . Killing of Candida albicans (1-3 blastospors/phagocyte) was seen after 2 h of incubation . The percentage of Listeria and Mycobacterium killed by phagocytes depended on pathogenicity of the tested strains . The bactericidal activity of phagocytes estimated in the XTT assay and by the CFU method was quite similar.

J Fr Ophtalmol, 1995, 18(12), 819 - 21
{In vitro changes induced by phenylmercury nitrate on fungi causing keratitis . Scanning electron microscopy study}; Duhamel C et al.; Antifungal activity of phenylmercury nitrate was studied on fungi from keratitis . Important deterioration and bursting were observed with 2 mg/100 ml phenylmercury nitrate concentration on Aspergillus flavus, Scedosporium and Candida albicans spores.

Braz Dent J, 1995, 6(2), 131 - 6
Oral manifestations of diabetes mellitus in controlled and uncontrolled patients; Quirino MR et al.; The authors studied the oral manifestations of a sample of 70 diabetic patients, divided into controlled and uncontrolled patients . Medical history and stomatological data were analyzed and diabetic controlled patients were matched to uncontrolled patients . The main symptoms observed were hyposalivation, taste alterations and burning mouth, with the main sign being parotid enlargement . The lesions observed were candidosis of the erythematous type and proliferative lesions both associated to the use of total prosthesis . No pathognomic lesions or alterations could be observed in relation to the disease . The frequency of carriers of Candida albicans and also the lesions observed could be compared to normal patients also using total dentures.

Mycopathologia, 1995-96, 132(3), 123 - 8
Synergy of fluconazole with macrophages for antifungal activity against Candida albicans; Garcha UK et al.; The possible between macrophages and fluconazole for antifungal activity against different isolates of C . albicans was studied . The susceptibility of C . albicans isolates to fluconazole (FCZ), when incubated in RPMI-1640 with 10% fetal bovine serum (FBS) and 10% fresh mouse serum (test medium, TM) was determined by using a quantitative culture methodology, Multiplication of isolate Sh27 was strongly inhibited by FCZ, even at 1.0 microgram/ml . However, FCZ even at 100 microgram/ml was not fungicidal . Resident murine peritoneal macrophages (MP) incubated for 48 h in RPMI-1640 + 10% FBS (tissue culture medium, TCM), then challenged with Sh27 in TM for 24 h, were fungistatic (20 +/ 9%, n = 4) . Cultured macrophages synergized with FCZ (10 micrograms/ml) for fungicidal activity when co-cultured with sh27 in TM for 24 h (46 +/ 8%) and for 48 h (74 +/ 5%), n = 3 . Macrophages and FCZ (10 micrograms/ml) could not synergize for significant killing of a less FCZ-sensitive C . albicans isolate 94-164 . Multiplication of a FCZ-resistant isolate (94-20) was not inhibited by FCZ at 10 micrograms/ml TM; however, macrophages and FCZ (10 micrograms/ml) could synergize for fungistatic (64%), but not fungicidal, activity.

Scand J Infect Dis, 1995, 27(4), 421 - 4
Fluconazole failure in two cases of disseminated candidosis; Jahnson L et al.; Amphotericin B has been the standard treatment of disseminated candidosis although the less toxic fluconazole tends to be used more frequently . In a recent report, no significant difference in the efficacy of candidaemia treatment was observed between fluconazole and amphotericin B . However, we report on 2 cases of invasive candidosis where fluconazole failed to eradicate Candida albicans although the isolates were susceptible to fluconazole in vitro . Pulsed-field gel electrophoresis was used to confirm the persistence of the same C . albicans strain after therapy failure in both patients.

Scand J Infect Dis, 1995, 27(4), 419 - 20
Candida albicans meningitis in a 27 weeks premature infant treated with liposomal amphotericin-B (AmBisome)
Jarlov JO, Born P, Bruun B.
We report a case of Candida albicans meningitis in a neonate born after 27 weeks of gestation . To the best of our knowledge this is the first report of a premature infant with Candida-meningitis treated with liposomal amphotericin B (AmBisome(R)) . The patient did not respond well to conventional Amphotericin B, but was successfully cured with Ambisome(R) . Liposomal amphotericin B was well tolerated and the baby recovered with a postinfectious hydrocephalus which necessitated a permanent ventriculo-peritoneal shunt . Six months after the infection the baby appears to have a near-normal cerebral development.

Scand J Infect Dis, 1995, 27(4), 391 - 5
Oral Candida albicans isolates with reduced susceptibility to fluconazole in Swedish HIV-infected patients; Chryssanthou E et al.; A total of 62 patients with HIV-related conditions were examined for clinical and mycological oral findings . Cultures from 51 patients were positive for yeasts and included 49 Candida albicans and 8 non-albicans isolates . Of patients with positive culture, 35% had pseudomembranous thrush . In vitro susceptibility testing of 49 C . albicans isolates revealed that the minimal inhibitory concentration for 50% of the strains (MIC50) was 2.0 mg/l for fluconazole, and the MIC50 was < or = 0.125 mg/l for both ketoconazole and itraconazole . Fluconazole resistance (MIC > or = 32.0 mg/l) was found for 14% of the C . albicans isolates tested . Two C . albicans isolates showed cross-resistance to ketoconazole and itraconazole . Associations between reduced susceptibility to fluconazole and low CD4+ cell counts, the length of time since the first AIDS-defining illness and the interval from the first fluconazole treatment, indirectly reflecting the total fluconazole exposure, were observed.

Folia Histochem Cytobiol, 1995, 33(3), 157 - 62
Changes in actin cytoskeleton are involved in the cytopathic action of Candida albicans upon human skin fibroblasts; Marewicz E et al.; The infection of human skin fibroblasts grown in culture with Candida albicans causes the death of infected cells, as assayed with the trypan blue exclusion test . The cell death is preceded by: (1) an attachment of germ tubes to the perinuclear surface of fibroblasts; (2) the disappearance of stress fibers and the disorganization of F-actin cytoskeleton and (3) cell rounding and detachment from the substratum . The results provide evidence for the role of F-actin cytoskeleton as a target in the cytopathic activity of C . albicans against human skin fibroblasts.

Microbiol Immunol, 1995, 39(7), 443 - 50
Biomaterial-associated infection with Candida albicans in mice; Rozalska B et al.; Candida yeasts are frequently isolated from patients with continuous ambulatory peritoneal dialysis peritonitis or other biomaterial-associated infections . The mouse model of candidal peritonitis was used to study the interaction of Candida cells with end-point attached heparinized polyethylene (H-PE) and with polymorphonuclear leukocytes (PMNs) or macrophages (M phi) . Two Candida strains differing in cell surface hydrophobicity and in expression of fibronectin (Fn) binding were used for the study . Cells of both Candida strains adhered at higher numbers to H-PE surfaces preadsorbed with Fn or with human dialysis fluid (HDF) than to non-modified H-PE, supporting a role of Fn in mediating adhesion . C . albicans 4016 cells expressing low hydrophobicity and low binding of soluble Fn demonstrated stronger adhesion to PMNs than the more hydrophobic C . albicans 3248 yeasts, which express high binding of soluble Fn . However, C . albicans 4016 cells were more resistant to phagocytic killing and were hardly eradicated in intraperitoneally infected mice . The animals depleted in PMNs by treatment with CY were neither able to eradicate C . albicans 3248 (rapidly eliminated by normal mice) nor C . albicans 4016 yeasts (with a tendency to persist in the tissues of normal mice).

J Fr Ophtalmol, 1995, 18(10), 614 - 6
{Palpebral mycosis}; Offret H et al.; A 47-year-old man presented with a necrotic and haemorrhagic ulceration of the lower eyelid . He had a-one-year history of secondary hepatocarcinoma . The patient underwent excision biopsy of the lesion . Histological examination showed fungal spores, which could be candida albicans.

Eur Arch Otorhinolaryngol, 1995, 252(7), 417 - 21
DNA amplification for the in vitro detection of Candida albicans in head and neck squamous cell carcinomas; Werner JA et al.; DNA was extracted from whole cells of Candida albicans and digested with HindIII restriction enzyme . After electrophoresis in a segment of the lane containing between 800 and 1200 base pairs (bp) of DNA fragments, a 1.1-kilobase (kb) fragment was found that hybridizes to biopsied tumor cells from head and neck squamous cell carcinomas (SCC) . From the nucleotide sequence of the putative gene locus, primers were synthesized for use in a polymerase chain reaction (PCR) with DNA extracted from 18 SCC of the upper aerodigestive tract . After 30 cycles of amplification all tumors were found to contain sufficient amplified DNA to be detected in polyacrylamide or agarose gels . In contrast, template DNA from lymph nodes and malignant lymphomas failed to generate positive signals under these conditions . However, samples of DNA obtained from head and neck SCC cells in vitro, Candida glabrata, and Candida parapsilosis after PCR were found to contain homologous sequences . Application of this technique to head and neck SCC biopsies may help to identify quickly the presence of concurrent candidal species.

Microbiol Immunol, 1995, 39(6), 405 - 9
Suppression of anti-Candida activity of murine neutrophils by progesterone in vitro: a possible mechanism in pregnant women's vulnerability to vaginal candidiasis; Nohmi T et al.; Sex steroid hormones were examined for their effect on mycelial growth of Candida albicans, and the inhibitory activity of casein-induced murine peritoneal neutrophils against mycelial growth of C . albicans was examined in vitro using a crystal violet staining method or a {3H}glucose incorporation method . Four steroid hormones, danazol, estradiol, estriol and testosterone had no effect on mycelial growth of C . albicans, but progesterone appeared to convert the growth form of C . albicans from hyphal to yeast . Danazol (10(-6) M) and progesterone (10(-5) M) suppressed anti-Candida activity of neutrophils of non-treated mice, while testosterone, estradiol, and estriol did not . The anti-Candida activity of neutrophils of estradiol-pretreated mice was clearly suppressed by progesterone even at 10(-6) M which corresponded to its plasma concentration in pregnant women in the third trimester . The physiological significance of this suppressive effect of progesterone was discussed in relation to the vulnerability of pregnant women to vaginal candidiasis.

Adv Perit Dial, 1995, 11, 277 - 80
Peritoneal membrane failure in children on peritoneal dialysis; Neiberger RE; Peritoneal dialysis (PD) success depends on adequate renal function replacement . Reports in adult dialysis populations indicate the risk of ultrafiltration failure (UF) increases with the time on dialysis . Type 1 UF is the most common . For children, dialysis modalities are temporizing measures until renal transplantation, considered optimal therapy for end-stage renal disease in children, can be performed . Children are frequently on dialysis for less than 2 years prior to transplantation . Thus if type 1 UF frequency increases with time on dialysis, it would be expected to occur less frequently in children, because they often are on dialysis for shorter periods . A retrospective chart review was performed to determine the cause of ultrafiltration failure in children; 172 children, mean age 8.0 +/- 5.5 (SD) years received a mean of 15 +/- 11.9 (SD) months of chronic PD; 39 patients received only continuous ambulatory peritoneal dialysis, 18 received only continuous cycling peritoneal dialysis, 22 received only tidal PD, and 94 received more than one type of PD . Ten patients (5.8%) developed type 2 UF failure as sequellae of atypical peritonitis, Candida albicans (6 patients), Mycobacterium foruitum (2), Achromatium spp . (1), and Pseudomonas aeruginosa (1) . There was no significant difference in time on dialysis for children who developed membrane failure . No patients with type 1 or type 3 UF could be identified . It appears that the causes of peritoneal membrane failure in children are different from those in adults.

Immunol Res, 1995, 14(2), 148 - 62
T helper cell dichotomy to Candida albicans: implications for pathology, therapy, and vaccine design; Romani L et al.; Acquired immunity to Candida albicans is believed to prevent mucosal colonization of adult immunocompetent individuals from progressing to symptomatic infection . Resistance to disease appears to correlate with the detection of delayed-type hypersensitivity responses in vivo and a T helper type 1 (Th1) cytokine secretion profile in vitro . Cellular immunodeficiency, particularly HIV infection, greatly increases the risk of mucosal infection, confirming that CD(4+)-cell-directed immunity is effective locally in controlling infectivity of the yeast . While Th1-type CD4+ cell activation resulting in phagocyte-dependent immunity clearly represents an important mechanism of anticandidal resistance, clinical observations suggest that Th2-type CD4+ cell reactivity may be triggered by Candida antigens in several disease states, including symptomatic infections and immunopathology . This may imply that a Th1-type pattern of reactivity characterizes the saprophytic yeast carriage and resistance to disease by healthy humans, whereas Th2-type responses would be mostly associated with pathology . Moreover, Candida-specific T helper responses, namely humoral and cell-mediated immunity, appear to be reciprocally regulated, as typically occurs in experimental models of parasitic and retroviral infection, where the Th1/Th2 paradigm of acquired immunity has been best characterized . Recent studies, besides providing direct evidence for the occurrence of cross-regulatory Th1 and Th2 responses in mice with candidiasis, emphasize the potential of cytokine/anticytokine therapy for recruiting Candida-specific responses toward protective, Th1-type CD4+ cell reactivity . At the same time, these studies call attention to the possible consequences of C . albicans infection for immunopathology, allergy, and coinfection.

Adv Exp Med Biol, 1995, 371A, 201 - 6
The leucocyte protein L1 (calprotectin): a putative nonspecific defence factor at epithelial surfaces; Brandtzaeg P et al.; The L1 protein occurs at high concentrations in neutrophils, monocytes, certain reactive tissue macrophages, squamous mucosal epithelia, and reactive epidermis . It constitutes in fact about 60% of the neutrophilic cytosol protein fraction . The two L1 chains (L1H and L1L) are referred to by a bewildering collection of names, various authors having different preferences (MRP-8 and MRP-14; CFA or calgranulin A and B) . The most recent proposal is calprotectin because of its calcium-binding properties and antimicrobial effect shown in vitro . L1 belongs to the S-100 protein family and may be involved in the regulation of keratinocyte proliferation and differentiation . It exists at high levels in blood and interstitial tissue fluid in several infectious, inflammatory, and malignant disorders, and it is released abundantly in foci of granulocytes and macrophages . The C-terminal sequence of the L1H chain has been shown to be identical to the N-terminus of peptides known as neutrophil immobilizing factors . Such an activity of L1 could be important for the accumulation of vital granulocytes, while L1 released from neutrophils, macrophages and epithelial cells might exert antimicrobial activity, perhaps by depriving microorganisms of zinc . The minimum inhibitory concentrations of L1 in vitro were found to be 4-32 mg/l for Candida albicans, 64 mg/l for Staphylococcus aureus, 64-256 mg/l for S . epidermidis, and 256 mg/ml for Escherichia coli and Klebsiella spp . Killing was observed at 2-4 times higher concentrations . In patients with HIV infection, those who developed oral candidiasis had significantly lower parotid L1 levels than those who did not (67 micrograms/l vs . 216 micrograms/l).

Ginecol Obstet Mex, 1995 Jan, 63, 15 - 8
{Itraconazole in the treatment of Candida vulvovaginitis in patients with type II diabetes mellitus (non-insulin dependent)}; Gonzalez Ortiz M et al.; The purpose of this study was to know the utility of traditional administration of itraconazole to vulvovaginitis by Candida in patients with diabetes mellitus . A randomized, single blind, controlled clinical assay, was carried out in 32 patients with diabetes mellitus type II and vulvovaginitis for Candida albicans . A study group was formed with 16 patients . They received 200 mg/day of itraconazole with breakfast for 3 days . The other 16 women were the control group . A good clinical response was obtained in 87.50% of the patients (p = 0.001) . Candida disappeared in 56.25% of the cases . General response showed medication failure in 43.75% . It is concluded that the traditional treatment of itraconazole for vulvovaginitis for Candida albicans in women with diabetes mellitus is a good choice to control the signs and symptoms, but it doesn't erradicate the fungus.

Microbiology, 1995 Jan, 141 ( Pt 1), 213 - 9
Adherence of Candida albicans to human salivary components adsorbed to hydroxylapatite; Cannon RD et al.; Colonization of the oral cavity by Candida albicans involves adherence of yeast cells to oral surfaces . An assay was developed to measure the attachment of C . albicans cells, metabolically labelled with {35S}methionine, to saliva-coated hydroxylapatite (SHA) beads--a model for the tooth surface . Using this assay approximately 1.26 x 10(6) C . albicans cells (50.5% of input cells) attached to the SHA beads (12 mg) . Different strains of C . albicans adhered to varying degrees to SHA beads, but in general adherence was promoted by growth of cells at 28 degrees C and by starvation of cells for glucose . Proteins in human whole, or parotid, saliva samples were fractionated by gel filtration (Sephacryl S-200 column) and fractions were adsorbed to hydroxylapatite beads . Fractions that contained proline-rich proteins or statherin promoted attachment of C . albicans ATCC 10261 cells . Neuraminidase treatment of SHA beads, but not of cells, significantly increased yeast cell binding to the SHA beads . Neuraminidase activity in the oral cavity may unmask glycoprotein receptors involved in the adhesion of C . albicans to saliva-coated surfaces.

Dermatology, 1995, 190(1), 39 - 42
Fungal infections in the Netherlands . Prevailing fungi and pattern of infection; Korstanje MJ et al.; BACKGROUND: Species of fungi have specific characteristics in geographic distribution, and they have a predilection for certain body areas . OBJECTIVE: To obtain information about the prevailing fungi and their pattern of infection in the Netherlands . METHODS: An analysis was made of the results of mycological examinations carried out in subjects referred to the mycological laboratory of the Department of Dermatology, University Hospital Leiden, the Netherlands, in the period of 1972-1992 . RESULTS: The feet, extremities and groin were most commonly infected . The feet and extremities were mainly infected with dermatophytes (Trichophyton rubrum), but in the groin Candida albicans accounted for 49.9% of the fungal infections . Onychomycosis ranked third in prevalence (17.3% of all fungal infections) . The main etiologic agent in the toenails was T . rubrum, but in fingernails C . albicans seems to be at least as important as T . rubrum . On the trunk (mainly the chest, especially in women) and buttocks C . albicans was again the main etiologic agent for fungal infections . Dermatophytes accounted for only 32.4% and 14.2% of the fungal infections on the buttock and trunk, respectively . On the buttock and trunk T . rubrum was the main etiologic agent as far as dermatophytes are concerned . On the trunk, Microsporum canis and M . ferrugineum were of some importance as well . The prevalence of tinea capitis was very low and accounted for only 0.7% of all fungal infections . CONCLUSION: Cutaneous candidosis accounted for 30.3% of all fungal infections and is therefore important . On the chest, buttocks, groin and finger-nails, the prevalence of C . albicans is higher than that of dermatophytes.

Arzneimittelforschung, 1995 Jan, 45(1), 84 - 7
Inhibition of Candida albicans adhesiveness to human buccal and vaginal cells by sub-inhibitory concentrations of rilopirox; Braga PC et al.; Candida albicans is an opportunistic dimorphic pathogenic yeast which is present on the human mucosal epithelial cell surface . Its adhesion is considered to be an important first step in colonization and in the subsequent symptomatic or asymptomatic infection of buccal or vaginal mucosa . Because the ability to adhere is an important element of the pathogenicity of Candida we investigated in this study the compared effects of sub-inhibitory concentrations (sub-MICs) of rilopirox (CAS 104153-37-9) with those of ciclopirox olamine (CAS 41621-49-2) in inhibiting Candida adhesion to human buccal (BEC) and vaginal cells (VEC) . Rilopirox is a new hydroxypyridone antimycotic agent with strong activity, especially against Candida albicans . There was a significant reduction in the mean number of Candida adhering to both buccal and vaginal cells with up to 1/8 MIC rilopirox for buccal and 1/16 MIC for vaginal cells, while for ciclopirox olamine reduction was significant up to 1/16 MIC for buccal and 1/8 MIC for vaginal cells . There were no significant differences in the dose-effect curves for BEC and VEC with either rilopirox and ciclopirox olamine, but on a molar basis, rilopirox was more active than ciclopirox olamine . The present in-vitro results support the developmental concept of an oropharyngeal and vaginal preparation of rilopirox . It can be expected that even sub-inhibitory concentrations of rilopirox exert an important additional effect in the treatment of oral and vaginal candidosis by impairing the pathogenic adhesion process of the fungus.

Infect Immun, 1995 Jan, 63(1), 280 - 8
Noninhibitory binding of human interleukin-2-activated natural killer cells to the germ tube forms of Candida albicans; Arancia G et al.; During incubation in vitro with yeast or germ tube forms of Candida albicans, only 2 to 6% of freshly isolated human natural killer (NK) cells (> 85% CD16+, CD56+, CD3-; < 15% CD3+; cytolytic for the NK-susceptible target K562 but not for the NK-resistant target DAUDI), were seen to interact with the fungal cells . As seen under the electron microscope, the contact area had a limited extent and was narrow, and neither the surface nor the intracytoplasmic organization of the NK cell was altered . In contrast, more than 30% of interleukin-2-activated NK (LAK) cells (> 96% CD16+, CD56+, CD3-; 1.5% CD3+; cytolytic for both K562 and DAUDI targets) interacted closely with the fungus . This interaction was particularly extensive with the surface of the fungal germ tube that was intimately enveloped by villous protrusions from the lymphocyte surface . The fungus-interacting LAK cell also showed a remarkable redistribution of surface microvilli and polarization of cytoplasmic organelles, such as the Golgi apparatus, centrioles, and granules, toward the area of fungal contact . Together with the elevated cytolytic potential against the K562 and DAUDI targets, all the morphological data suggested the presence of a potentially active lytic machinery in the fungus-interacting LAK cell . Nonetheless, two independent assays for anticandidal activity did not show consistent killing or fungal growth inhibition by either fresh NK or LAK cells . While offering direct evidence of the strong interaction between human LAK cells and the germ tubes, precursors of tissue-invasive hyphal forms of C . albicans, our observations also suggest that this interaction may not be sufficient to kill the fungus or arrest its growth.

J Biochem (Tokyo), 1995 Jan, 117(1), 54 - 8
Palmitoyl lysozyme-induced stabilization of PE (phosphatidylethanolamine) liposomes and their interaction with Candida albicans; Lee EO et al.; The interaction with fungal cells of dioleoylphosphatidylethanolamine liposomes coated with palmitoyl lysozyme was investigated using lysozyme as a stabilizer for an unstable bilayer as well as a recognizer for a specific target cell . Lysozyme, which interacts with chitin in the fungal cell wall, lyses chitin and kills the cells, was acylated with N-hydroxysuccinimide ester of palmitic acid (NHSP) and incorporated into the lipid bilayer . Lysozyme was optimally modified when the ratio of NHSP to lysozyme was 15 at pH 8.0 . Modification of lysozyme was detected by SDS-PAGE, and its molecular weight was about 1,500 greater than that of the intact lysozyme at the optimal ratio of NHSP to lysozyme . The activity of palmitoyl lysozyme toward glycolchitin was reduced to 35% of that of intact lysozyme . Both lysozyme and palmitoyl lysozyme had antifungal activities, but palmitoyl lysozyme was more effective than intact lysozyme against Candida albicans . The minimal molar ratio of palmitoyl lysozyme to phosphatidylethanolamine required to form stable liposomes was 2.4 x 10(-4), and the optimal ratio was about 2.4 x 10(-3) . The percentage survival of cells treated with the inserted palmitoyl lysozyme was lower than that of cells treated with free palmitoyl lysozyme . These findings suggest that palmitoyl lysozyme-incorporated liposomal membrane is more effectively adsorbed by Candida albicans than free palmitoyl lysozyme is.

J Antimicrob Chemother, 1995 Jan, 35(1), 155 - 9
Improved method of determining the susceptibility of Candida albicans to fluconazole; Polanco AM et al.; The MICs of fluconazole for four reference strains of Candida albicans were determined in one laboratory by the macrobroth dilution method recommended by the National Committee for Clinical Laboratory Standards (NCCLS) and by a modified version of the method which incorporated microbroth dilution, RPMI 1640 medium supplemented with glucose and the reading of MIC endpoints by spectrophotometry . The results obtained by the two methods were within one two-fold dilution . In addition, the susceptibilities of 58 clinical isolates of C . albicans to fluconazole were determined in two laboratories by the modified NCCLS method . Close interlaboratory agreement was observed, 98.2% of MICs varying by no more than one two-fold dilution.

J Antimicrob Chemother, 1995 Jan, 35(1), 103 - 14
Emergence of azole drug resistance in Candida species from HIV-infected patients receiving prolonged fluconazole therapy for oral candidosis; Johnson EM et al.; We examined the effect of different fluconazole treatment regimens on the emergence of azole drug resistance among Candida species recovered from the mouths of 54 HIV-infected individuals . Patients were assigned to one of three treatment groups depending on their history of oral candidosis and fluconazole use . Mouthwashes obtained at regular intervals were cultured and isolates identified using standard methods . Antifungal broth micro-dilution tests were performed to determine IC30s of fluconazole and ketoconazole . Sixty-four Candida albicans isolates from 20 patients with no evidence of oral candidosis who had not received fluconazole all had IC30s of < or = 4 mg/L . Thirty-four (83%) of 41 C . albicans isolates from ten patients receiving intermittent, short-term fluconazole treatment for oral candidosis had IC30s of < or = 4 mg/L, but only two isolates (5%) had IC30s > or = 64 mg/L . In contrast, 26 (40%) of 65 C . albicans isolates from 15 patients given long-term fluconazole (50-200 mg/day or 150 mg/week) were classified as resistant having IC30s of fluconazole of > or = 64 mg/L . Ten of these 26 fluconazole-resistant isolates were susceptible to ketoconazole with IC30s of < or = 4 mg/L suggesting azole drug cross-resistance is not inevitable . Tests on multiple colonies from individual isolation plates showed that it was not unusual to obtain differing IC30 values, indicating that a sweep inoculum is essential if resistance is to be detected . Nine (60%) of the 15 patients given long-term fluconazole harboured isolates of C . albicans that were resistant to fluconazole at some time during the study period . All had low CD4 counts and were approaching the final stage of their illness . Three patients on long-term treatment had resistant organisms at the outset of the study; in the remainder, resistant strains emerged during the study period . In six of the nine cases, emergence of resistance in vitro correlated with persistent clinical signs of oral infection . Thirty-six isolates of Candida species other than C . albicans were also recovered from patients receiving long-term fluconazole and 29 (81%) of these had IC30s of > or = 64 mg/L . Our experience with C . albicans in patients with HIV infection, suggests that the long-term azole drug use may be an important factor in the development of fluconazole resistance as such resistance was rare and transient in patients on intermittent short-term treatment.

Mol Microbiol, 1995 Jan, 15(1), 39 - 54
Candida albicans ALS1: domains related to a Saccharomyces cerevisiae sexual agglutinin separated by a repeating motif; Hoyer LL et al.; Transfer of budding Candida albicans yeast cells from the rich, complex medium YEPD to the defined tissue culture medium RPMI 1640 (RPMI) at 37 degrees C and 5% CO2 causes rapid onset of hyphal induction . Among the genes induced under these conditions are hyphal-specific genes as well as genes expressed in response to changes in temperature, CO2 and specific media components . A cDNA library constructed from cells incubated for 20 min in RPMI was differentially screened with yeast (YEPD)- and hyphal (RPMI)-specific probes resulting in identification of a gene expressed in response to culture conditions but not regulated by the yeast-hyphal transition . The deduced gene product displays significant identity to Saccharomyces cerevisiae alpha-agglutinin, encoded by AG alpha 1, an adhesion glycoprotein that mediates mating of haploid cells . The presence of this gene in C . albicans is curious since the organism has not been observed to undergo meiosis . We designate the C . albicans gene ALS1 (for agglutinin-like sequence) . While the N- and C-termini of the predicted 1260-amino-acid ALS1 protein resemble those of the 650-amino-acid AG alpha 1, ALS1 contains a central domain of tandem repeats consisting of a highly conserved 36-amino-acid sequence not present in AG alpha 1 . These repeats are also present on the nucleotide level as a highly conserved 108 bp motif . Southern and Northern blot analyses indicate a family of C . albicans genes that contain the tandem repeat motif; at least one gene in addition to ALS1 is expressed under conditions similar to those for ALS1 expression . Genomic Southern blots from several C . albicans isolates indicate that the number of copies of the tandem repeat element in ALS1 differs across strains and, in some cases, between ALS1 alleles in the same strain, suggesting a strain-dependent variability in ALS1 protein size . Potential roles for the ALS1 protein are discussed.

Acta Derm Venereol, 1995 Jan, 75(1), 75 - 8
Human papilloma virus infection among women attending an STD clinic correlated to reason for attending, presence of clinical signs, concomitant infections and abnormal cytology; Voog E et al.; The purpose of this study was to demonstrate the prevalence of cervical human papilloma virus (HPV) infection correlated to reason for attending an STD clinic, presence of clinical signs of HPV infection, concomitant infection and abnormal cytology . Samples from the cervical canals of 588 consecutive women attending the STD clinic, Department of Dermato-Venereology, Sahlgrenska Hospital, Gothenburg, were taken with a Cytobrush for detection of HPV DNA with the dot blot/Southern-blot technique . Visible condylomata, i.e . filiform or papular condylomata, were registered . Acetic acid test and colposcopy were not routinely performed . Cytological examination was performed as well as isolation of Chlamydia trachomatis on Mc Coy's cells and culture on Sabouraud agar for Candida albicans . The prevalence of HPV DNA was 8% (48/588) . In the group of 233 women attending because of concern about HPV infection, 94 (40%) had visible signs of HPV infection and 30 (13%) were positive for HPV DNA in the cervix . In 355 women attending for other reasons, such as discharge, pruritus or STD check-up, 4 (1%) had visible signs of HPV infection and 18 (5%) were HPV DNA positive . Of 98 women with visible signs of vulvar/vaginal HPV infection, 33 (34%) were HPV-positive in the cervix with a commercial Southern-blot test . Of 490 patients without visible signs of HPV infection, 15 (3%) were HPV-positive in the cervix . In the group of HPV-positive women a positive culture for Candida was demonstrated in 26% (11/43), Compared to 16% (79/504) of the HPV-negative women.(ABSTRACT TRUNCATED AT 250 WORDS)

Clin Infect Dis, 1995 Jan, 20(1), 77 - 83
Isolation of fluconazole-resistant Candida albicans from human immunodeficiency virus-negative patients never treated with azoles; Goff DA et al.; Isolation of fluconazole-resistant strains of Candida species from human immunodeficiency virus (HIV)-infected patients after repeated or continuous courses of treatment has been reported with increasing frequency . During 1991-1992, MICs of fluconazole for 139 Candida albicans isolates from our institution were bimodally distributed: 102 strains were susceptible (MICs, < or = 4 micrograms/mL) and 37 were resistant (MICs, > or = 8 micrograms/mL) . There was incomplete cross-resistance between fluconazole and ketoconazole or miconazole, and there was no cross-resistance between azoles and amphotericin B or flucytosine . Twenty of the 37 fluconazole-resistant strains were isolated from 17 HIV-negative patients, some with systemic infections, who had never been treated with azoles . There were no differences in characteristics or risk factors for those patients as compared with those for an equal number of HIV-negative patients from whom fluconazole-susceptible strains were isolated . Among patients with systemic infection, 6 (50%) of 12 with infection caused by fluconazole-resistant strains survived and 11 (69%) of 16 with infection caused by fluconazole-susceptible strains survived (P = .54) . Survival was not found to be related to treatment regimen, but the number of patients was small . The emergence of fluconazole-resistant C . albicans among HIV-negative patients never exposed to azoles is of concern.

J Oral Pathol Med, 1995 Jan, 24(1), 32 - 6
Biotypes of oral Candida albicans isolates in human immunodeficiency virus-infected patients from diverse geographic locations; Tsang PC et al.; Oral Candida albicans isolates from HIV-infected individuals in Hong Kong, Australia, Germany and England were characterised using a biotyping system based on enzyme profiles, carbohydrate assimilation patterns and boric acid resistance of the yeasts . A total of 44 biotypes were found amongst the 117 oral C . albicans isolates examined . The major biotype A1R accounted for 17.9% of all isolates while the second commonest biotype was A1S (11.1% of isolates) . Whereas these two biotypes were isolated from all the regions studied, there were a number of other biotypes unique to individual countries . The data indicate that there are many different sub-strains of oral C . albicans in HIV-infected patients, some of which are globally prevalent . However, further work is required to ascertain the diversity of oral C . albicans biotypes, if any, in health and diseasePIP: 11-96% of patients with HIV infection develop oral candidosis at some point during the progression of HIV infection to AIDS . In the early stages of HIV infection, the development of oral candidosis is highly predictive of worsening immunodeficiency . Despite its importance as a sentinel opportunistic infection in HIV disease, however, little is known about the epidemiology of the major etiological agent, Candida albicans, associated with the disease . The authors conducted a study to identify the different biotypes of C . albicans isolated from oral samples of HIV-infected patients from Hong Kong, Australia, Germany, and England, and to gain insight into their geographic distribution . 33 isolates from Hong Kong, 37 from Australia, 30 from Germany, and 17 from England were characterized using a biotyping system based upon enzyme profiles, carbohydrate assimilation patterns, and boric acid resistance of the yeasts . 44 biotypes were identified . A1R and A1S were the two major biotypes, accounting for 17.9% and 11.1% of all isolates, respectively, isolated from all the regions studied . Some other biotypes were unique to individual countries . This study therefore found that there are many different sub-strains of oral Candida albicans in HIV-infected patients, some of which are globally prevalent .

FEMS Immunol Med Microbiol, 1995 Jan, 10(2), 151 - 5
Functional impairment of rat Kupffer cells induced by aflatoxin B1 and its metabolites; Cusumano V et al.; Contamination of food with mycotoxins is a major health problem . Impairment of several immune functions has been repeatedly reported in animals fed with contaminated fodder . Since the liver is a major target of toxicity by aflatoxins, the effects of aflatoxins B1, and its hepatic metabolites Q1 and M1 on Kupffer cell function was investigated in vitro . Aflatoxin B1 induced significant (P < 0.05) inhibition of phagocytosis, intracellular killing of Candida albicans, and intrinsic anti-Herpes virus activity at concentrations as low as 0.01 pg ml-1 . Aflatoxin Q1 and M1 had similar effects on phagocytosis and microbicidal activity, but were two- to ten-fold less potent than aflatoxin B1.

Exp Nephrol, 1995 Jan-Feb, 3(1), 58 - 60
Antineutrophil cytoplasmic antibody-positive sera inhibit candidacidal activity of granulocytes; Bartunkova J et al.; Antineutrophil cytoplasmic antibodies (ANCA) are suspected of being involved in the pathogenesis of tissue injury in systemic vasculitis . We have investigated the effect of 10 sera from 8 patients with ANCA-associated diseases on the capacity of neutrophils derived from healthy persons to kill ingested Candida albicans . ANCA-containing sera inhibited candidacidal activity by 55-80% in comparison to control sera . This phenomenon could lead to the depression of antimicrobial resistance of patients with ANCA and could be involved in the pathogenesis of granuloma formation.

Ann Pharmacother, 1995 Jan, 29(1), 10 - 5
Salivary concentrations of ketoconazole and fluconazole: implications for drug efficacy in oropharyngeal and esophageal candidiasis; Force RW et al.; OBJECTIVE: To determine whether salivary concentrations of ketoconazole and fluconazole may explain the apparent disparity between in vitro activity and clinical efficacy observed with these drugs . DESIGN: Healthy subjects received a single oral dose of ketoconazole 400 mg or fluconazole 100 mg in a randomized, crossover fashion . Saliva was collected at 0, 1, 2, 3, 6, 12, and 24 hours . Blood samples were obtained at 2 and 24 hours . Salivary concentrations and plasma concentrations for each drug were determined by HPLC . Minimum inhibitory concentration (MIC) testing was determined in triplicate on 6 clinical isolates of Candida albicans, and times over the median MIC values were calculated . PARTICIPANTS: Eight subjects completed the study . RESULTS: The mean (+/- SD) peak salivary concentration for ketoconazole was 0.119 +/- 0.050 microgram/mL at 3 hours; no subject had a detectable ketoconazole salivary concentration at 24 hours . At 2 and 24 hours, mean ketoconazole plasma concentrations were 7.64 +/- 3.87 and 0.11 +/- 0.05 microgram/mL, respectively . The saliva to plasma concentration ratio at 2 hours was 0.01 . The mean peak salivary concentration of fluconazole was 2.56 +/- 0.34 microgram/mL at 3 hours . At 24 hours, the mean salivary concentration was 1.44 +/- 0.33 microgram/mL . At 2 and 24 hours, mean fluconazole plasma concentrations were 4.39 +/- 3.33 and 3.72 +/- 2.83 micrograms/mL, respectively . The saliva to plasma concentration ratio at 2 hours was 0.55 . Median MIC values were 0.0625 microgram/mL (range 0.0313-0.125) for ketoconazole and 0.25 microgram/mL (range 0.125-0.5) for fluconazole . Calculated times over which ketoconazole and fluconazole exceeded the median MICs in saliva were approximately 13 and greater than 24 hours, respectively . CONCLUSIONS: After a single oral dose, fluconazole achieved higher salivary concentrations than did ketoconazole . This may explain the increased clinical efficacy of fluconazole in the treatment of oropharyngeal-esophageal candidiasis when compared with ketoconazole.

Jpn J Antibiot, 1995 Jan, 48(1), 140 - 5
{In vitro antifungal activities of lanoconazole against stock cultures and clinical isolates of Candida albicans}; Niwano Y et al.; In vitro antifungal activities of lanoconazole (LCZ) against 9 stock cultures and 10 clinical isolates of Candida albicans were determined using three different testing media, Sabouraud's glucose broth (SGB), Sabouraud's glucose agar (SGA) and casitone agar (CA) . MIC values of LCZ against both the stock cultures and clinical isolates measured on CA distributed in a range of 0.63-5 micrograms/ml . The values were 8-64-fold lower than those obtained in SGB and on SGA . MIC ranges and the geometric mean MIC values of LCZ against stock cultures were virtually the same as those against clinical isolates, no matter which assay techniques were used . These results suggest that there is no difference in the LCZ susceptibility between stock cultures and clinical isolates of C . albicans, although anti-Candida activity of LCZ was determined to be greater on CA than that in SGB or on SGA.

Antimicrob Agents Chemother, 1995 Jan, 39(1), 40 - 4
Antifungal susceptibility testing of isolates from a randomized, multicenter trial of fluconazole versus amphotericin B as treatment of nonneutropenic patients with candidemia . NIAID Mycoses Study Group and the Candidemia Study Group; Rex JH et al.; The antifungal susceptibilities of 232 pathogenic blood stream Candida isolates collected during a recently completed trial comparing fluconazole (400 mg/day) with amphotericin B (0.5 mg/kg of body weight per day) as treatment for candidemia in the nonneutropenic patient were determined both by the National committee for Clinical Laboratory Standards M27-P macrobroth methodology and by a less cumbersome broth microdilution methodology . For amphotericin B, M27-P yielded a very narrow range of MICs (0.125 to 1 microgram/ml) and there were no susceptibility differences among species . For fluconazole, a broad range of MICs were seen (0.125 to > 64 micrograms/ml), with characteristic MICs seen for each species in the rank order Candida albicans < C . parapsilosis approximately equal to C . lusitaniae < C . glabrata approximately equal to C . krusei approximately equal to C . lipolytica . The MIC distribution for C . tropicalis was bimodal and could not be ranked . Both microdilution MICs were within one tube dilution of the M27-P MIC for > 90% of isolates with amphotericin B and for > or = 77% of isolates with fluconazole . For both methods, elevated MICs did not predict treatment failure . In the case of amphotericin B, the MIC range was too narrow to permit identification of resistant isolates . In the case of fluconazole, MICs for isolates associated with failure to clear the bloodstream consistently were equivalent to the median MIC for the given species . Successful courses of therapy were seen with four isolates from four patients despite MICs of > or = 32 micrograms/ml . As MICs obtained by M27-P and similar methods correlate with responsiveness to fluconazole therapy in animal models and in AIDS patients with oropharyngeal candidiasis, the lack of correlation in this setting suggests that the MICs for these isolates are at or below the relevant fluconazole breakpoint for this dose of fluconazole and patient setting and that host factors such as failure to exchange intravenous catheters were more important than MIC in predicting outcome.

Cah Anesthesiol, 1995, 43(1), 13 - 9
{Beware of yeast septicemia in intensive care medicine}; Dhahri MA et al.; A retrospective study of 24 cases of systemic candidosis observed in a polyvalent intensive care unit over a 6.5 yr period (1987-1993) led to some constatations: an increasingly high incidence of this type of septicaemia (up to 27.5% of all septicaemias), large responsibility of skin saprophytes Candida ( > 62% vs 21% from intestinal Candida albicans), frequent diagnostic difficulties, and a fatal outcome in 7/24 patients (mainly from severe causal illness) . In order to improve the prognosis, a more systematic and often empirical resort to fungicidal agents could be justified whenever patients with very severe surgical or medical conditions develop a protracted fever of unclear origin.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 83 - 5
In vitro susceptibility to itraconazole and fluconazole of switch phenotypes of Candida albicans serotypes A and B, isolated from immunocompromised hosts; Velegraki A; Data from 78 oropharyngeal samples obtained from severely immunocompromised individuals, have shown that the B serotype and the stipple phenotype of Candida albicans isolates have significantly higher minimum inhibitory concentration values towards itraconazole and fluconazole, as compared with the corresponding serotype A phenotype (P < 0.05) . The epidemiological significance of the phenotype-dependent drug susceptibility patterns is discussed.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 77 - 80
Genetic differentiation of Candida albicans strains by mixed-linker polymerase chain reaction; McCullough MJ et al.; Restriction fragment length polymorphisms (RFLPs) of C . albicans were visualized using a mixed-linker polymerase chain reaction (PCR) and a C . albicans-specific primer (C . albicans 1059-bp species-specific repeat sequence) . The method produced 10-14 bands of between 200 and 1400 bp on agarose gel electrophoresis and ethidium bromide staining . It gave comparable results to Southern blot hybridization with good reproducibility and the time required for the production of typing profiles was reduced to less than 2 days.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 53 - 8
Solubilization and identification of essential functional groups of Candida albicans oxidosqualene cyclase; Carrano L et al.; The enzyme properties and location of essential functional groups of solubilized oxidosqualene cyclase of Candida albicans have been studied . We show that the C . albicans enzyme is much more heat-labile compared with Saccharomyces cerevisiae and rat liver cyclases, requires a histidyl residue for enzyme activity, contains an essential thiol residue either close to or in the active site and exhibits a carbocationic mechanism for catalysis, as the enzyme-bound substrate protects the enzyme from inactivation by a site-directed inactivator.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 49 - 52
Development of Candida albicans and C . albicans/Escherichia coli/Bacteroides fragilis intraperitoneal abscess models with demonstration of fungus-induced bacterial translocation; Sawyer RG et al.; The development of models of both Candida albicans and mixed C . albicans/Escherichia coli/Bacillus fragilis peritonitis in immunologically normal mice are described, each with significant mortality and intra-abdominal abscess formation . C . albicans inoculated alone induced bacterial translocation into abscesses, and the addition of bacteria reduced the number of, but did not eliminate . C . albicans in abscesses . There was no synergy seen between fungi and bacteria in terms of either morbidity or mortality.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 33 - 7
A longitudinal study of the change in resistance patterns and genetic relationship of oral Candida albicans from HIV-infected patients; McCullough M et al.; Candida albicans is an important opportunistic pathogen in the oral cavity of HIV-infected patients . C . albicans was isolated repeatedly over a period of 30 months from the mouth-swills of five males with category IV HIV infection who suffered from recurrent pseudomembranous oral candidosis and were being treated with fluconazole . C . albicans was identified by the germ tube test, carbon assimilation by the API 20C test (bioMerieux sa, Marcy, l'Etoile, France) and morphology on cornmeal Tween 20 agar . Sensitivity testing was performed with a disk diffusion method using neosensitabs (Rosco Diagnostica, Taasrup, Denmark) and the broth microdilution assay using RPMI 1640 medium . Isolates from the five patients taken between 14 and 30 months apart (mean = 23 months) showed the development of resistance to several antifungal agents, notably fluconazole, miconazole and itraconazole by the disc diffusion method and markedly higher minimum inhibitory concentrations (MICs) for fluconazole by the broth microdilution assay . Genetic relatedness of the C . albicans strains was assessed using the mixed-linker polymerase chain reaction method . Four patients had isolates which were genetically identical, while one patient had isolates of varying genetic type . This study has shown that in HIV-infected patients with recurrent oral pseudomembraneous candidosis, the development of drug resistance to antifungal agents may either be due to increased resistance of a single strain or replacement with a more resistant strain.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 1 - 8
Induction of candidal vaginitis in diabetic mice and attempts to prevent the infection; Segal E et al.; In the present study on the induction and prevention of candidal vaginal infection in diabetic mice, ICR female mice were rendered diabetic by intraperitoneal injections of 160 mg kg-1 streptozotocin (SZ) which induced a stable non-lethal diabetic state apparent 2-7 days post-SZ injection . Diabetic mice and non-diabetic controls were inoculated intravaginally with 10(7)-10(10) Candida albicans yeasts . Infection was evaluated by the number of infected mice 3 days post-inoculation and the rate of clearance of infection during a 35-day follow-up . All C . albicans doses investigated induced vaginal infection in the diabetic mice, while in the non-diabetic controls only 40% of those inoculated with the lower (10(7) CFU) dose developed infection . The yeast load in the diabetic mice was 10-fold higher than in the controls and the duration of infection was longer . In an attempt to prevent infection intravaginal treatment with 5-40 mg of a chitin derivative (CSE) mouse-1 was given prior and pre- and post-inoculation with C . albicans . The optimal CSE treatment regimen was both pre- and post-administration of 10 or 20 mg CSE mouse-1 which significantly reduced the number of infected animals and also the duration of infection.

Mycoses, 1995 Jan-Feb, 38(1-2), 37 - 40
Discrimination of strains of Candida albicans isolated from deep and superficial sites by resistotyping; Hunter PR; Resistotyping was used to characterise 106 strains of Candida albicans classified as to whether they came from deep or superficial infections . The data were analysed by logistic regression analysis . There was a statistically significant difference between the two groups of strains . After removal of variables by the log-likelihood method four resistotyping agents were found to predict source of strain: boric acid, benzalkonium chloride, malachite green and mercurochrome . Whilst there were phenotypic differences between strains isolated from deep and superficial sites it is not clear whether this strain represents different strain types or strain histories.

Mycoses, 1995 Jan-Feb, 38(1-2), 29 - 36
Validity of contour-clamped homogeneous electric field electrophoresis as a typing system for Candida albicans; Sangeorzan JA et al.; Instigated by an increase in serious human Candida infections and aided by advances in technology, there has been renewed interest in the study of the epidemiology of fungal infections . Among the newer techniques available, contour-clamped homogeneous electric field (CHEF) electrophoresis has shown great promise as a tool for typing strains of Candida albicans . However, few studies have addressed the reproducibility of the preparatory and electrophoretic methods . Through a series of analyses on clinical isolates of C . albicans, we were able to demonstrate that (a) sample preparation induced no appreciable artifacts in CHEF banding patterns; (b) the electrophoretic patterns were reproducible over time; (c) changes in colony morphology were not associated with changes in the electrophoretic pattern, and (d) the method was more sensitive than restriction enzyme analysis (REA) for demonstrating strain differences . CHEF electrophoresis is a sensitive and reproducible tool for the study of Candida epidemiology . Further use and study of this methodology is warranted.

Mycoses, 1995, 38 Suppl 1, 33 - 9
{Changes in the fungal spectrum of dermatomycoses}; Tietz HJ et al.; The spectrum of aetiologic agents isolated from 3607 patients suspicious for dermatomycosis being in the care of the Berlin Charite Clinics was analysed . Identification of dermatophytes and moulds were performed conventionally . For the identification of yeasts biochemical and genetic methods were used . Among the dermatophytes in comparison of present with previous incidence rates changes can be observed . Opportunistic yeasts are recognized in increasing importance . Five fungal species are forming a stable base of aetiologic agents of dermatomycoses, i.e . Trichophyton rubrum, T . mentagrophytes, Candida albicans, C . parapsilosis and Trichosporon cutaneum, completed by increasing incidence of Microsporum canis, T . mentagrophytes var . granulosum and T . tonsurans.

Mycoses, 1995, 38 Suppl 1, 14 - 21
{Post-antibiotic effect and post-exposure polyene antagonism of azole antimycotics in Candida albicans: dependency on lipophilia}; Scheven M et al.; With the lipophilic azoles itraconazole (ICZ), ketoconazole (KCZ), and miconazole (MCZ) two effects, occurring in parallel, on Candida albicans were observed: Firstly, these azoles caused a growth inhibition which persisted for at least 24 hours (post-antibiotic effect, found regularly with KCZ and MCZ, with ICZ only occasionally) . Furthermore, the fungicidal activity of amphotericin B (AMB, 1 mg/1) after exposure to the azoles was reduced . In contrast, to this, fluconazole (FCZ) produced neither of these effects . Additional experiments indicate that both actions of the three lipophilic azoles may be related to their noncovalent binding to lipophilic cytoplasmatic components of the yeast cells . In the case of fluconazol such bonds seem to be much weaker . Presumably, the amount of the relatively hydrophilic fluconazole, which will be bound to the cell, is too low as to produce long lasting post-exposure effects like those caused by the lipophilic azoles.

Arch Dermatol Res, 1995, 287(5), 448 - 51
Ketoconazole in atopic dermatitis: therapeutic response is correlated with decrease in serum IgE; Back O et al.; The prevalence of specific IgE antibodies to the yeasts Pityrosporum orbiculare and Candida albicans was investigated in adult patients with atopic dermatitis (AD) or with seborrhoeic dermatitis and in healthy controls by means of the radioallergosorbent test (RAST) . Of 63 AD patients, 28 (44%) had IgE antibodies to P . orbiculare and 21 (33%) to C . albicans . This is highly significant, since no antibodies were found in sera from other patients or controls . With the intention to treat, 20 patients with AD and a positive RAST to P . orbiculare were given ketoconazole 200 mg daily for 2 months and 200 mg twice weekly for further 3 months . The clinical scores improved during treatment with a reduction in the levels of specific IgE to P . orbiculare and total serum IgE . However, there were no correlations between clinical score and serum levels of P . orbiculare-specific IgE . C . albicans-specific IgE, on the other hand, correlated both with clinical score and with total serum IgE.

Am J Reprod Immunol, 1995 Jan, 33(1), 86 - 93
Effects of endometrial serpin-like proteins on immune responses in sheep; Skopets B et al.; PROBLEM: The uterine milk proteins (UTMP) are a pair of related glycoproteins that are the major secretory products of the endometrium of the pregnant ewe . UTMP are members of the serpin superfamily of serine protease inhibitors but have no known antiprotease activity . One possible role for UTMP is to inhibit uterine immune responses--UTMP inhibit mitogen and mixed lymphocyte-induced proliferation of peripheral blood lymphocytes, natural killer (NK) cell activity and abortion caused by NK cell activation . Present objectives were to further evaluate the lymphocyte-inhibitory activity of UTMP and test whether UTMP modify immune responses in vivo . METHOD: One experiment demonstrated that UTMP inhibited antigen-induced lymphocyte proliferation induced by Candida albicans extract . In another experiment, ewes were immunized with OVA mixed with 3.75 mg/ml of UTMP or ovine serum albumin (OSA control) . Injections of 1 mg OVA+UTMP or OSA in incomplete adjuvant were administered 6 wk later . Titers of antibody to OVA were lower (P < 0.001) for ewes administered UTMP than for ewes administered OSA . Effects of UTMP on delayed hypersensitivity reactions were evaluated in three experiments using skin-fold thickness assays . RESULTS: UTMP did not inhibit the increase in skin-fold thickness caused by PHA and Mycobacterium tuberculosis but rather tended to increase the response to PHA . CONCLUSION: Results strengthen the thesis that UTMP are physiologically relevant immunoregulatory molecules . Nonetheless, effects on skin-fold responses indicate that actions of UTMP can be more complex than would be predicted based on the proteins only having a single biological effect.

Cell Biol Int, 1995 Jan, 19(1), 65 - 9
A GCN-like response in Candida albicans; Pereira SA et al.; The control of amino acid and purine biosynthesis in the yeast Saccharomyces cerevisiae is mediated by the transcriptional activator GCN4 . We previously identified the presence of two putative GCN4 responsive elements (GCREs) in the promoter sequence of the Candida albicans ARO3 gene, which encodes an enzyme in the aromatic amino acid pathway . We now show that amino acid deprivation results in a dramatic rise in the steady-state level of ARO3-specific mRNA, indicative of a GCN-like pathway in C . albicans.

Crit Rev Oral Biol Med, 1995, 6(2), 147 - 60
Oral leukoplakia; Sciubba JJ; Leukoplakia has evolved as a clinico-pathologic concept over many years, with the current clinical designation being accepted worldwide . Reflective of the biology of leukoplakia is the concept of cellular atypia and epithelial dysplasia . Adding to a better understanding of leukoplakia in general has been the definition of relevant clinical subsets which, in some cases, includes etiology (snuff), while in other cases a verrucous clinical appearance will suggest a more aggressive anticipated behavior pattern . Tobacco usage, in many of its forms, remains the prime etiologic factor; however, other considerations also apply . More recently, the potential etiologic role of Candida albicans has been stressed, as well as its possible role in carcinogenesis . So-called oral hairy leukoplakia has been defined in relation to a possible Epstein-Barr viral infection, usually in the immunosuppressed patient . Other viruses, human papilloma virus in particular, have been implicated in leukoplakia, while genetic alterations involving tumor suppressor elements (p53) have also been investigated . Finally, the management of this common condition remains a variable and includes local, topical, and systemic therapies such as anti-oxidants, carotenoids, and retinoids.

Res Microbiol, 1995 Jan, 146(1), 73 - 83
Antimicrobial activity of 9-oxo and 9-thio acridines: correlation with interacalation into DNA and effects on macromolecular biosynthesis; Cremieux A et al.; The antimicrobial activity of several new 9-acridinones and 9-thioalkylacridines towards Escherichia coli, Staphylococcus aureus, Mycobacterium smegmatis and Candida albicans was investigated . Minimal inhibitory, bactericidal and fungicidal concentrations were determined using a microplate assay which enabled inhibitory, bactericidal and fungicidal indices to be calculated . These indices facilitated structure/activity relationship studies . DNA-intercalating capability and DNA supercoiling inhibitory effects as well as inhibitory effects on macromolecular synthesis were determined . Results showed that intercalation into DNA, which is the mechanism of action usually postulated for acridines, cannot be correlated with the properties examined . However, inhibition of RNA synthesis may be involved in the antimicrobial activity of the drugs.

Gynecol Obstet Invest, 1995, 39(1), 67 - 9
In situ mycotoxin production by Candida albicans in women with vaginitis; Shah DT et al.; The virulence attributes of Candida albicans in cases of mucocutaneous disease have not been identified . Based on the recent finding that C . albicans is able to produce an immunosuppressive mycotoxin, gliotoxin, we analyzed vaginal samples of 3 women severely symptomatic for vaginal candidiasis and found that they contained significant levels of gliotoxin . Three control women who were not colonized with C . albicans showed no gliotoxin in vaginal samples . These findings raise the possibility that gliotoxin may play a role in the virulence of C . albicans.

Acta Otorhinolaryngol Belg, 1995, 49(3), 251 - 5
Nosocomial pansinusitis in orotracheally intubated critically ill patients; Spapen H et al.; Nosocomial pansinusitis (N P) is most often described in nasotracheally intubated patients with craniocerebral or facial trauma . We retrospectively reviewed its occurrence and complications in the course of prolonged mechanical ventilation in orotracheally intubated patients without maxillofacial or cranial injuries . N P was deemed to be present when (1) CT scan showed opacification and/or air-fluid levels in the maxillary, ethmoid and sphenoid sinuses and (2) aspiration of both maxillary sinuses yielded pus, cultures of which revealed a high concentration of micro organisms . Nosocomial pneumonia and bacteremia were considered related to the N P if the organisms found in the sinus were identical to those recovered from the blood and/or the bronchi . During an 18-month study period, 38 cases of sinusitis were diagnosed . N P was present in 13 patients: 18 organisms (12 Gram-negative, 5 Gram-positive and 1 Candida albicans) were isolated . Pneumonia occurred in 8 patients, 6 with multi-resistant Pseudominas aeruginosa and 2 with methicillin resistant Staphylococcus aureus . Pseudomonas aeruginosa was isolated from the blood, lung and sinus in two patients . This study demonstrates that N P is relatively frequent in patients requiring long-term mechanical ventilation, even in the absence of predisposing factors . N P in these patients is mostly monomicrobial with multi-resistant Pseudomonas aeruginosa and Staphylococcus aureus as the main causative agents.

J Immunol, 1994 Dec 15, 153(12), 5643 - 9
Candidacidal mechanisms in the human neonate . Impaired IFN-gamma activation of macrophages in newborn infants; Marodi L et al.; We studied the interaction between Candida albicans and mononuclear phagocytes derived from cord blood . In the presence of normal serum, the extent of phagocytosis and killing of candida by monocyte-derived macrophages was equivalent for newborns and adults . In the absence of serum both phagocytosis and killing by macrophages were reduced by half, but cord and adult cells were still equivalent . Mannosylated BSA and mannan inhibited ingestion of unopsonized candida by macrophages, suggesting a role for the mannose receptor . Exposure of cord and adult macrophages to IFN-gamma (10-500 U/ml) gave quantitatively different results in Candida killing, as well as in release of superoxide anion (O2-) . Maximal increase in these functions with adult macrophages was achieved with 100 U/ml IFN-gamma . No enhancement with cord macrophages could be detected after treatment with 100 U/ml, and at 500 U/ml there was still significantly lower killing and O2- release compared with adult cells . Defective up-regulation of O2- release was also present in cord monocytes exposed to IFN-gamma on day 0 . Studies of the surface expression of IFN-gamma receptors using a "nonblocking" mAb against the IFN-gamma receptor revealed a comparable number of receptors on cord and adult monocytes . When blocking Abs were used, however, there was a three times higher number of positive cells in cord monocytes . Specific binding of 125I-IFN-gamma to cord monocytes and macrophages was also higher compared with adult cells . These data suggest that neonatal macrophages have a normal capacity to ingest and kill both opsonized and unopsonized Candida but cannot be fully activated by IFN-gamma, a finding that could not be attributed to lower expression of IFN-gamma receptors on the neonatal cells.

Mol Gen Genet, 1994 Dec 15, 245(6), 716 - 23
Molecular cloning and analysis of CDC28 and cyclin homologues from the human fungal pathogen Candida albicans; Sherlock G et al.; In the budding yeast Saccharomyces cerevisiae, progress of the cell cycle beyond the major control point in G1 phase, termed START, requires activation of the evolutionarily conserved Cdc28 protein kinase by direct association with G1 cyclins . We have used a conditional lethal mutation in CDC28 of S . cerevisiae to clone a functional homologue from the human fungal pathogen Candida albicans . The protein sequence, deduced from the nucleotide sequence, is 79% identical to that of S . cerevisiae Cdc28 and as such is the most closely related protein yet identified . We have also isolated from C . albicans two genes encoding putative G1 cyclins, by their ability to rescue a conditional G1 cyclin defect in S . cerevisiae; one of these genes encodes a protein of 697 amino acids and is identical to the product of the previously described CCN1 gene . The second gene codes for a protein of 465 residues, which has significant homology to S . cerevisiae Cln3 . These data suggest that the events and regulatory mechanisms operating at START are highly conserved between these two organisms.

Biochem Biophys Res Commun, 1994 Dec 15, 205(2), 1079 - 85
ACPR, a STE12 homologue from Candida albicans, is a strong inducer of pseudohyphae in Saccharomyces cerevisiae haploids and diploids; Singh P et al.; ACPR from Candida albicans encodes a protein antigenically related to the secretory acid proteinase of this yeast . Its amino terminal domain is highly similar to the amino terminal, DNA-binding domain of STE12 of Saccharomyces cerevisiae . STE12 is involved in mating of haploids and in pseudohyphae formation in diploids . ACPR, or its DNA-binding domain swapped into STE12, can support pseudohyphae formation in S . cerevisiae diploids . However, unlike STE12, these constructs affect the budding pattern and induce pseudohyphae formation in S . cerevisiae haploids as well, and this induction is independent of the nitrogen status of the medium . ACPR appears to be a stronger inducer of pseudohyphae than STE12 and is likely to be involved in the formation of pseudohyphae and hyphae in C . albicans.

Science, 1994 Dec 9, 266(5191), 1723 - 6
Suppression of hyphal formation in Candida albicans by mutation of a STE12 homolog; Liu H et al.; A Candida albicans gene (CPH1) was cloned that encodes a protein homologous to Saccharomyces cerevisiae Ste12p, a transcription factor that is the target of the pheromone response mitogen-activated protein kinase cascade . CPH1 complements both the mating defect of ste12 haploids and the filamentous growth defect of ste12/ste12 diploids . Candida albicans strains without a functional CPH1 gene (cph1/cph1) show suppressed hyphal formation on solid medium . However, cph1/cph1 strains can still form hyphae in liquid culture and in response to serum . Thus, filamentous growth may be activated in C . albicans by the same signaling kinase cascade that activates Ste12p in S . cerevisiae; however, alternative pathways may exist in C . albicans.

J Trauma, 1994 Dec, 37(6), 944 - 9
Monocytes overcome lymphocyte dysfunction in injured adults with elevated Candida antigen titers; Sweeney JF et al.; OBJECTIVE: Severely injured adults with elevated Candida antigen titers have increased mortality from sepsis, in part because of known neutrophil (PMN) dysfunction . Since PMN function is modulated by monocytes and lymphocytes, this study was undertaken to determine the ability of monocytes and lymphocytes isolated from injured adults with elevated Candida antigen titers to activate the anticandidal function of normal PMNs . METHODS: Lymphocytes with or without monocytes, isolated from 18 injured adults with elevated titers, were cultured in the presence or absence of heat-killed Candida albicans for 48 hours . Culture supernatants were harvested, diluted 1:40, 1:160, and 1:640, and tested for the ability to stimulate the anticandidal function of normal PMNs using an 3H-glucose incorporation assay . Monocytes and lymphocytes isolated from nine volunteers were studied for comparison . RESULTS: Supernatants of lymphocytes from healthy volunteers that were cultured with heat-killed C . albicans significantly augmented normal PMN anticandidal function . Supernatants of lymphocytes from injured adults with elevated titers that were cultured with heat-killed C . albicans did not significantly augment normal PMN anticandidal function . Supernatants of monocytes or lymphocytes from both groups of patients were able to upregulate PMN anticandidal function . CONCLUSIONS: Lymphocytes from injured adults with elevated Candida antigen titers are defective in their ability to stimulate PMN anticandidal function . Monocytes from these patients can respond to Candida exposure and overcome the lymphocyte functional defect seen in injured patients with elevated titers.

J Infect Dis, 1994 Dec, 170(6), 1566 - 9
Karyotyping of Candida albicans isolates obtained longitudinally in women with recurrent vulvovaginal candidiasis; Vazquez JA et al.; Ten women with recurrent vulvovaginal candidiasis (RVVC) due to Candida albicans were followed for a mean of 35.3 months, and 81 vaginal isolates were evaluated by pulsed-field gel electrophoresis for strain delineation . The initial strain of C . albicans isolated was unique to each patient; in addition, in 8 women, only 1 strain type of C . albicans was identified throughout follow-up . In 1 patient, 3 strains of C . albicans were identified over a 27-month period and in another, 2 strains were recovered over a 30-month period . Two pairs of women shared identical strains of C . albicans . These results confirm the enormous diversity of strain types of C . albicans and demonstrate the persistence of colonization with the same strain over prolonged periods despite therapy . Results also support the concept of recurrent vaginitis being due to vaginal relapse or endogenous reinfection with the identical strain.

J Infect Dis, 1994 Dec, 170(6), 1557 - 65
Response of human polymorphonuclear leukocytes and monocytes to Trichosporon beigelii: host defense against an emerging opportunistic pathogen; Lyman CA et al.; To further understand human host defenses against Trichosporon beigelii, functional responses were investigated of polymorphonuclear leukocytes (PMNL) and elutriated human monocytes (EHM) to this opportunistic fungal pathogen . There was significantly less PMNL phagocytosis (P < .001) and killing (P < .001) of T . beigelii isolates than of Candida albicans . However, levels of superoxide anions generated by PMNL in response to T . beigelii and C . albicans were comparable . Pretreatment of PMNL with granulocyte colony-stimulating factor or interferon-gamma (IFN-gamma) did not significantly enhance fungicidal activity . Killing of T . beigelii by EHM also was significantly impaired compared with killing of C . albicans (P < .001) . However, pretreatment of EHM with macrophage colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, or IFN-gamma all resulted in enhanced fungicidal activity . Thus, phagocytosis and killing of T . beigelii by PMNL and EHM are significantly less efficient than that of C . albicans . However, monocytes may be more important in the control of Trichosporon species than previously shown.

Arch Surg, 1994 Dec, 129(12), 1227 - 32
Glucocorticoid receptor blockade reverses postinjury macrophage suppression; Cech AC et al.; OBJECTIVES: To study the effects of the stress-induced surge of endogenous glucocorticoids on macrophage function and the role of inhibiting glucocorticoids with a receptor antagonist, mifepristone (RU 486) . DESIGN: One hundred thirty female Swiss-Webster mice were randomly assigned to either injury by femur fracture or uninjured anesthesia control in this intervention study . SETTING: A university-based surgical laboratory and animal facility . INTERVENTION: Injured mice were randomized to receive either the glucocorticoid receptor antagonist mifepristone (10 mg/kg by oral gavage) or its vehicle . Mifepristone or its vehicle were given either 2 hours before or 2 hours after the injury . MAIN OUTCOME MEASURES: Peritoneal macrophages were harvested 24 hours after the injury . Macrophages were assayed for the stimulated (phorbol myristate acetate, 1 microgram/mL) production of superoxide anion, secretion of interleukin-6, tumor necrosis factor alpha, and prostaglandin E2 in response to endotoxin (lipopolysaccharide at 10 micrograms/mL) and killing of Candida albicans . RESULTS: Pretreatment with mifepristone significantly prevented or reduced suppression of several macrophage functions following injury, including superoxide production and C albicans killing . Treatment after the injury preserved only C albicans . Mifepristone failed to block the increased secretion of prostaglandin E2 after injury . CONCLUSION: Pretreatment with mifepristone before an injury prevented suppression of several macrophage functions . Further studies are required on the effects of glucocorticoid inhibition on other aspects of the immune and metabolic responses to injury to define the potential clinical applications of mifepristone trauma.

Infect Immun, 1994 Dec, 62(12), 5353 - 60
A mannoprotein constituent of Candida albicans that elicits different levels of delayed-type hypersensitivity, cytokine production, and anticandidal protection in mice; Mencacci A et al.; To identify major immunogenic constituents of Candida albicans, the effect of a mannoprotein fraction (MP-F2) on the elicitation of a delayed-type hypersensitivity (DTH) reaction, cytokine production, and protection from a virulent Candida challenge in a mouse candidiasis model was studied . In mice immunized with whole cells of a low-virulence strain of C . albicans and thus protected against a challenge with a highly virulent strain of this fungus, MP-F2 was able to elicit a strong DTH response that was accompanied by splenocyte proliferation in vitro in the presence of Candida antigen . The supernatants of MP-F2-stimulated splenocyte cultures contained gamma interferon (IFN-gamma, a typical CD4+ T helper-1 (Th1) cytokine, but no interleukin-4, (IL-4), a typical CD4+ Th2 cytokine . IFN-gamma was produced by CD4+ cells, and its level could be greatly increased by the addition of anti-IL-4 or, mostly, anti-IL-10 antibodies to the CD4+ cell cultures . Upon a suitable schedule of immunization, MP-F2 was also able to induce a vigorous DTH response in Candida-uninfected mice, a response that could be efficiently transferred into naive recipients by CD4+ cells from the spleens of MP-F2-immunized mice . The immunization described above also conferred to mice a low degree of protection against a virulent Candida challenge, both in terms of median survival time and in the number of Candida cells in the kidney . However, while DTH induction by MP-F2 was as strong as that induced by whole cells, MP-F2-induced protection was significantly weaker than that conferred by Candida whole-cell immunization . Mice immunized with either MP-F2 or Candida whole cells had an inverted ratio between the number of CD4+ splenocytes producing IFN-gamma and that of cells producing IL-4, compared with nonimmunized animals . However, the number of IL-4-producing CD4+ cells was significantly higher in MP-F2-vaccinated, weakly protected mice than in Candida whole-cell-vaccinated, highly protected animals . Overall, our data suggest that the MP-F2 fraction contains one or more major immunogens of C . albicans which are capable of interfering with the balance of CD4+ Th1 and Th2 responses that is so critical in the outcome of host-Candida relationship and are thus potentially relevant in the mechanisms of Candida-specific DTH regulation and protection.

Clin Immunol Immunopathol, 1994 Dec, 73(3), 344 - 9
Effect of abscess fluid supernatants on the kinetics of Candida albicans growth; Radke LL et al.; Abscess fluid supernatants have been found to inhibit microbial growth, an effect that appears to be due to a protein originating in the cytoplasm of neutrophils . The antimicrobial activity of calprotectin, the responsible protein, is reversible by the addition of zinc to the culture medium . To more carefully analyze this type of antimicrobial activity supernatants of fluids from experimental subcutaneous Candida albicans abscesses in mice were studied to determine how they affect the in vitro growth kinetics of C . albicans . The abscess fluid supernatants inhibited growth in a dose-dependent fashion and more at early times than at later ones . Instability of the abscess fluid antimicrobial activity did not appear to explain the timing of the growth inhibition . A marked lengthening of the lag phase of growth was observed in cultures containing the supernatants (from approximately 6 hr in the control cultures to 15-20 hr in those with the supernatants) . On the other hand, the abscess fluid supernatants had only minimal effects on the generation times of actively proliferating C . albicans yeast cells . In addition, these supernatants did not appear to significantly affect the percentage of inoculated organisms undergoing cell division, as determined by a limiting dilution assay . Therefore, these results indicate that abscess fluid supernatants extend the lag phase of C . albicans growth, an effect similar to that seen with zinc-deprived organisms.

Acta Odontol Scand, 1994 Dec, 52(6), 335 - 45
Evaluation of the antimicrobial effects of sodium benzoate and dichlorobenzyl alcohol against dental plaque microorganisms . An in vitro study; Ostergaard E; Evaluation of antimicrobial agents is based on in vivo and in vitro studies . The minimum inhibitory concentrations (MICs) of sodium benzoate and dichlorobenzyl alcohol to 115 strains of plaque microorganisms were determined by a broth-dilution method . Sodium benzoate did not inhibit growth of any gram-positive cocci (MIC > 106,590 microM) . MICs for Porphyromonas gingivalis and two strains of Treponema socranskii were 26,650 microM . The MIC of dichlorobenzyl alcohol to the reference strain of Actinobacillus actinomycetemcomitans was 723 microM and to P . gingivalis, two strains of T . socranskii, and Candida albicans 1,446 microM . MICs for other organisms were 2,892 to 5,784 microM . Saliva samples from 10 volunteers, collected at various times after toothbrushing with a dentifrice containing 10% sodium benzoate and 0.3% dichlorobenzyl alcohol, were analyzed gas-chromatographically . Immediately after toothbrushing mean levels of sodium benzoate and dichlorobenzyl alcohol were 372,626 microM and 7,529 microM, respectively . After 5 min mean levels were 38,700 microM and 734 microM . In conclusion, the concentrations of both antimicrobials dropped rapidly during the first 30 min, but for 5-10 min they were high enough to inhibit growth of potential periodontal pathogens.

Microbiology, 1994 Dec, 140 ( Pt 12), 3319 - 28
Occurrence of chromosome rearrangements during the fusion process in the imperfect yeast Candida albicans; Suzuki T et al.; Auxotrophic derivatives of three strains of the pathogenic yeast Candida albicans of different origins, including 1006 derived from CBS5736, A5153 derived from FC18 and NARA2 derived from NUM961, were used in spheroplast fusion experiments . The DNA content of the prototrophic fusion product obtained following fusion between strains 1006 and A5153 approximated to the sum of those of the parents, but was variable when NARA2 was used as the parent for fusion . Chromosome-sized DNA molecules of the fusion derivatives were separated by pulsed-field gel electrophoresis to examine whether either or both of the chromosome-sized DNA molecules of each parent were transferred into the fusion derivatives . In the fusion derivatives obtained following fusion between strains 1006 and A5153, nearly the full complement of chromosomes was shown to be transferred, but partial transfer of chromosomes occurred in the fusion derivatives that were obtained following fusion between strains NARA2 and A5153 . Results indicated that chromosome loss also occurred when these two strains were fused . Variations in the size of R chromosomes, the rDNA-containing chromosomes, were observed in all fusion derivatives tested, indicating high-frequency recombination between R chromosomes during the fusion process.

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