Chemotherapy, 1994 Mar-Apr, 40(2), 124 - 35 Influence of membrane composition of various parasites on the inhibitory activity of a series of bipyridyl analogues; Seydel JK et al.; In a previous report the dependence of the antimycoplasmal activity of bipyridyl analogues on the presence of Cu+ ions has been shown . The inhibitory activity of these compounds has now been studied against Escherichia coli mycobacteria and Candida albicans in the absence and presence of Cu2+ ions using growth kinetic techniques . It was found that the inhibitory activity against E . coli increases in the presence of Cu2+ ions . In contrast, no additional effect of Cu2+ ions is observed for the inhibitory activity against mycobacteria . Some of the derivatives show very promising activity even against Mycobacterium avium strains . More complicated is the effect of Cu2+ ions on the inhibitory activity of the derivatives against C . albicans . For isoquinolones the observed delay in onset of inhibitors is reduced and no influence on the inhibitory activity is observed . The addition of Cu2+ ions to the phenanthrolines leads in contrast to a decrease in antifungal activity . The possible influence of membrane properties of the studied microorganism on the effect of Cu2+ ions is discussed. Cell Immunol, 1994 Mar, 154(1), 99 - 108 Effect of the psychoactive metabolite of marijuana, delta 9-tetrahydrocannabinol (THC), on the synthesis of tumor necrosis factor by human large granular lymphocytes; Kusher DI et al.; The natural killer cell (NK)/3polymorphonuclear neutrophil axis has recently been identified to be important in early defense against the opportunistic fungi, Candida albicans . Repression of this system is therefore likely to contribute to susceptibility to opportunistic infections . delta 9-Tetrahydrocannabinol (THC), an active constituent of marijuana, has been reported to be immunosuppressive at concentrations that exceed attainable plasma levels . In this report, we examine the possibility that human large granular lymphocytes (LGL) can be immunosuppressed by exposure to THC at physiologically relevant concentrations and probed two functions associated with LGL, i.e., cytokine production and tumoricidal activity . We find that these low levels of THC inhibit tumor necrosis factor-alpha (TNF) induction from LGL by C . albicans and are dependent upon THC dose (0.005-5.0 micrograms/ml) and length of exposure (0.05-3.0 hr) . Northern blot analysis indicates that the downregulation of TNF production from LGL by THC resides at the mRNA level . Moreover, exposure of LGL to physiological THC concentrations (0.01-2.0 micrograms/ml) diminishes their cytolytic activity against K562 tumor cells. Infect Immun, 1994 Mar, 62(3), 968 - 73 Structural modification of cell wall mannans of Candida albicans serotype A strains grown in yeast extract-Sabouraud liquid medium under acidic conditions; Kobayashi H et al.; The cell wall mannans of two Candida albicans serotype A strains, NIH A-207 and J-1012 (abbreviated as A and J strains, respectively), cultured in yeast extract-Sabouraud liquid medium at pH 2.0, contained neither a phosphate group nor a beta-1,2-linked mannopyranose unit (H . Kobayashi, P . Giummelly, S . Takahashi, M . Ishida, J . Sato, M . Takaku, Y . Nishidate, N . Shibata, Y . Okawa, and S . Suzuki, Biochem . Biophys . Res . Commun . 175:1003-1009, 1991) . In this study, the mannans obtained from A and J strains grown in pH 2.0 medium (abbreviated as mannans A2 and J2, respectively) exhibited quite different reactivities against rabbit anti-C . albicans and anti-Saccharomyces cerevisiae sera compared with those of mannans from the corresponding strains cultured in conventional medium at pH 5.9 (abbreviated as mannans A and J, respectively) . Namely, mannans A2 and J2 lost reactivity against the former serum but reacted with the latter serum to a higher extent than mannans A and J . In order to account for these difference in more detail, mannans A2 and J2 were subjected to acetolysis . Elution profiles of the acetolysates were completely different from those of acetolysates obtained from mannans A and J reported in our previous papers . The 1H nuclear magnetic resonance spectra of the oligosaccharides from mannans A2 and J2 obtained by this procedure indicate that the side chains are composed of alpha-linked mannopyranose units densely linked to the alpha-1,6-linked backbone . The long side chains containing one alpha-1,3-linked mannopyranose unit are markedly increased. Infect Immun, 1994 Mar, 62(3), 892 - 6 Immunoglobulin A (IgA), IgA1, and IgA2 antibodies to Candida albicans in whole and parotid saliva in human immunodeficiency virus infection and AIDS; Coogan MM et al.; Human immunodeficiency virus (HIV)-infected individuals are predisposed to recurrent oral candidiasis, and, although it has been assumed that this is because of deficient mucosal immune responses, this has not been properly established . The present study aimed to compare the concentrations and secretion rates of immunoglobulin A (IgA) and IgA subclass antibodies to Candida albicans in whole and parotid saliva samples from HIV-infected patients, AIDS patients, and control subjects . Levels of IgA antibody to Candida species in whole saliva were higher in the HIV group than in the controls and were highest in the AIDS group (P < 0.05) . In parotid saliva, the mean antibody levels were significantly greater in HIV-positive patients than in controls (P < 0.05) but fell to lower levels in the AIDS group . The secretion rates of Candida antibodies in parotid saliva were reduced in AIDS patients compared with HIV patients . The specific activities of the IgA antibodies and both subclasses were significantly higher in the HIV and AIDS patients than in the controls in both whole and parotid saliva (P < 0.05) . Antibody levels were significantly correlated with the numbers of Candida organisms isolated from saliva (P < 0.05) . These results suggest clear differences in salivary antibody profiles among HIV-infected . AIDS, and control subjects and are indicative of a response to antigenic challenge by infecting Candida species . No obvious defect in the mucosal immune response in the HIV or AIDS groups that might account for the increased prevalence of candidiasis was apparent. Infect Immun, 1994 Mar, 62(3), 828 - 36 Identification and characterization of a Candida albicans-binding proteoglycan secreted from rat submandibular salivary glands; Hoffman MP et al.; A previously identified Candida albicans-binding glycoprotein secreted from rat submandibular glands (RSMG) has been further purified from an aqueous RSMG extract by ion-exchange chromatography and gel filtration . Biochemical analysis of the glycoprotein revealed high levels of uronic acid and sulfate, suggesting that it was a proteoglycan . Its amino acid and carbohydrate compositions were similar to those observed for other proteoglycans and differed significantly from those of RSMG mucin, the major secretory glycoprotein of RSMG . In addition, the apparent molecular weight of the glycoprotein was reduced following treatment with either chondroitinase ABC or heparitinase, demonstrating the presence of chondroitin sulfate and heparan sulfate . On the basis of its structure and anatomical source, the glycoprotein is referred to as submandibular gland secreted proteoglycan 1 (SGSP1) . SGSP1 also binds monoclonal antibody 1F9, which recognizes the human blood group A carbohydrate epitope found on RSMG mucin . Hence, SGSP1 appears to be a hybrid molecule with carbohydrate structures found in both proteoglycans and RSMG mucin . Enzymatic digestion of SGSP1, followed by its interaction with a radiolabelled C . albicans strain in a filter-binding assay, demonstrated that binding to this strain appears to be mediated primarily via the heparan sulfate side chains of SGSP1 and not via the blood group A oligosaccharide. Infect Immun, 1994 Mar, 62(3), 1125 - 7 Ca ions stabilize the binding of complement factor iC3b to the pseudohyphal form of Candida albicans; Spotl L et al.; The pseudohyphal form of Candida albicans is able to bind iC3b . This may play an important role in the pathogenesis of disseminated candidiasis and, in particular, in adherence to endothelium, protection against complement action, and iron acquisition from erythrocytes . Here we report that Ca2+ ions are required to maintain stable binding of iC3b to C . albicans pseudohyphae. Infect Immun, 1994 Mar, 62(3), 1064 - 9 Mechanisms by which Candida albicans induces endothelial cell prostaglandin synthesis; Filler SG et al.; One strategy for improving resistance to opportunistic pathogens is to determine host cellular responses during the invasion process and upregulate those responses that are relevant to host defense mechanisms . Within this context, we have shown previously that invasion of endothelial cells by Candida albicans in vitro causes increased production of prostaglandins . As a prerequisite for modulating endothelial cell prostaglandin production, we now characterize the mechanisms through which this process occurs . Endothelial cell invasion by C . albicans appeared to stimulate the conversion of arachidonic acid into prostaglandins by upregulating the synthesis of endothelial cell cyclooxygenase and increasing the activity of the endothelial cell phospholipase . The enhanced activities of these two enzymes were independent of calphostin C-sensitive protein kinase C and resulted in the increased production and extracellular secretion of prostaglandin I2 (PGI2), PGF2 alpha, and PGE2 . The secretion of these prostaglandins had no effect on the amount of endothelial cell injury induced by C . albicans . The role of the increased prostaglandin secretion by endothelial cells is likely related to modulation of the leukocyte response at the candida-leukocyte-endothelial cell interface. Infect Immun, 1994 Mar, 62(3), 1032 - 8 Effects of preinduced Candida-specific systemic cell-mediated immunity on experimental vaginal candidiasis; Fidel PL Jr et al.; It has been postulated that systemic cell-mediated immunity (CMI) is an important host defense factor against recurrent vaginal infections caused by Candida albicans . Using an estrogen-dependent murine model of vaginal candidiasis, we have previously shown that mice inoculated vaginally with C . albicans acquire a persistent vaginal infection and develop Candida-specific Th1-type systemic CMI . In the present study, experimental vaginitis was monitored in the presence of preinduced systemic Candida-specific CMI . Mice immunized systemically with C . albicans culture filtrate antigens (CaCF) in complete Freund's adjuvant (CFA) had Th1-type reactivity similar to that of vaginally infected mice . CaCF given to mice intravenously induced Candida-specific suppressor T (Ts) cells . Mice preimmunized with CaCF-CFA and given a vaginal inoculum of C . albicans had positive delayed-type hypersensitivity (DTH) reactivity from the time of vaginal inoculation through 4 weeks . Conversely, mice infected in the presence of Ts cells had significantly reduced DTH responses throughout the 4-week period in comparison with naive infected mice . However, the presence of Th1-type Candida-specific DTH cells or Ts cells, either induced in mice prior to vaginal inoculation or adoptively transferred at the time of inoculation, had no effect on the vaginal Candida burden through 4 weeks of infection . A similar lack of effects was obtained in animals with lower Candida population levels resulting from a reduction in or absence of exogenous estrogen . These results suggest that systemic Th1-type CMI demonstrable with CaCF is unrelated to protective events at the level of the vaginal mucosa. Infection, 1994 Mar-Apr, 22(2), 132 - 6 Correlation between antifungal susceptibility testing of Candida isolates from patients with HIV infection and clinical results after treatment with fluconazole; Ruhnke M et al.; In an open-label controlled study 23 HIV-infected patients (CDC IV A-E) with documented oropharyngeal candidosis were treated with 100 mg fluconazole orally over 5 days (53 episodes; 1-6 treatments/patient) . Efficacy data were compared with a control group of 21 patients who received treatment for 10-21 days with 100 mg fluconazole for candidosis . Candida isolates were repeatedly recovered from patients before and after treatment with fluconazole and antifungal susceptibility testing (microbroth-dilution) was done . Inoculum size, medium pH, incubation time and temperature were standardized . Up to 85% of patients responded to therapy clinically and mycologically . Candida albicans was the most important yeast (86%) isolated from cultures of oral washings . In 90% of C . albicans isolates MIC to fluconazole were low (< or = 1.56 mg/l) . Primary resistance to fluconazole was not seen, but secondary resistance occurred in two cases clinically and in vitro (MIC > or = 25 mg/l) . Short treatment for 5 days was as successful as for 10 to 21 days without leading to significantly more recurrences of oral candidosis in these patients . Selection of Candida spp . other than C . albicans (e.g . Candida krusei, Torulopsis glabrata) under repeated fluconazole treatment occurred rarely . One patient developed clinical signs of chronic recurrent candidiasis, where only C . krusei could be cultured repeatedly. Infection, 1994 Mar-Apr, 22(2), 124 - 31 In vitro susceptibility and sterol biosynthesis of Candida albicans strains after long-term treatment with azoles in HIV-infected patients; Hundt W et al.; Over a period of 6 to 24 months a long term follow up of the in vitro antifungal susceptibility of 306 Candida albicans strains from 49 HIV-infected patients was performed . Using a microdilution test, the strains were tested against the azoles ketoconazole, itraconazole and fluconazole . The susceptibility range for fluconazole was between 1 and 128 mg/l, for itraconazole between 0.015 and 32 mg/l and for ketoconazole between 0.007 and 16 mg/l . 11.7% of the strains showed elevated IC30-values against ketoconazole, 9.1% against itraconazole and 10.1% against fluconazole . Sterol biosynthesis was examined by thin layer chromatography in 18 less sensitive strains and nine sensitive strains in the presence of ketoconazole . The proportion of ergosterol in the presence of ketoconazole at a concentration of 0.003 mg/l varied between 7.6% and 21.1% in sensitive strains and between 11.1% and 86.6% in less sensitive strains . In resistant control strains the proportion of ergosterol was 73% and 94.2%, respectively . Without ketoconazole the ergosterol proportion was > 85% in all strains . There was a significant correlation between the IC30-values and the inhibition of ergosterol biosynthesis (p = 0.05). J Ethnopharmacol, 1994 Mar, 42(1), 25 - 9 Cytotoxic and antimicrobial screening of selected terpenoids from Asteraceae species; Villarreal ML et al.; Twelve pure compounds originally obtained through a systematic chemotaxonomical study with Mexican plants of the Asteraceae, were subjected to a cytotoxic and in vitro antimicrobial screening . Three different cell lines in culture (KB, KB-VI and P388) were used in the cytotoxicity assay, while antimicrobial activity was tested against Gram-positive and Gram-negative bacteria, as well as Candida albicans . Of the twelve terpenoids tested, only taraxasterol showed antimicrobial activity against Staphylococcus aureus . The significant cytotoxic activity exhibited by five sesquiterpene lactones, and the moderate cytotoxicity of an eudesmane, is discussed. J Pharm Pharmacol, 1994 Mar, 46(3), 186 - 91 Effect of hydroxypropyl-beta-cyclodextrin on the antimicrobial action of preservatives; Lehner SJ et al.; The interaction between hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) and several preservatives with different chemical structures was investigated in aqueous solution . Complex stability constants of the 1:1 complexes were calculated from differential spectra . Using the serial dilution test the antimicrobial activities of the preservatives and their complexes against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans were tested and MIC values determined . For highly water-soluble substances like thimerosal and bronopol, low or no inactivation was found; the more lipophilic substances, such as the phenolic compounds, showed strong inactivation when used in combination with HP-beta-CyD . The loss in activity by complex formation correlated with the bound fraction, thus suggesting that the appropriate antimicrobial substance for the preservation of cyclodextrin solutions can be selected according to the results of this study. Nutrition, 1994 Mar-Apr, 10(2), 151 - 4 Altered gut barrier function to Candida during parenteral nutrition; Pappo I et al.; We hypothesize that catheter-related sepsis with Candida during total parenteral nutrition (TPN) is caused by Candida translocation from the gut . Fifty male Sabra rats weighing 330 +/- 40 g were randomized into four groups and put into metabolic cages: group 1 (n = 16), nonoperated free-feeding controls; group 2 (n = 10), infused with normal saline and free feeding; group 3 (n = 14), infused with TPN solution for a total of 36 kcal and 1.5 g g protein.100 g-1 body wt.day-1;group 4 (n = 10), same TPN regimen as group 3 but also receiving oral and intravenous antibiotics . On day 7, all animals received 1.5 x 10(10) viable Candida albicans CBS 562 cells by gavage, and 24 h later, the number of Candida colony-forming units in blood, mesenteric lymph nodes, and kidneys was determined . No growth of Candida was detected in group 1 or group 2 . Positive Candida cultures were found in the blood, mesenteric lymph nodes, and kidneys of groups 3 and 4, although levels reached statistical significance only for mesenteric lymph nodes in group 3 . Because Candida growth occurred exclusively in groups receiving TPN and bowel rest, we conclude that altered gut-barrier function to Candida occurs during TPN and speculate that Candida sepsis during TPN might be the result of Candida translocation from the gut due to the combination of high-density Candida colonization and favorable local conditions in the gut induced by TPN and bowel rest. J Oral Pathol Med, 1994 Mar, 23(3), 130 - 2 The effect of exposure to chlorhexidine gluconate in vitro and in vivo on in vitro adhesion of Candida albicans to buccal epithelial cells from diabetic and non-diabetic subjects; Darwazeh AM et al.; The effect of 0.2% chlorhexidine gluconate on in vitro adhesion of Candida albicans to buccal epithelial cells (BEC) was studied in 12 healthy subjects and 12 patients with diabetes mellitus . Exposure of BEC for one minute with 0.2% chlorhexidine gluconate in vitro, or by rinsing the mouth in vivo, resulted in a significant reduction in candidal adhesion to BEC in both diabetic and non-diabetic subjects and between both groups . In addition to the known fungicidal effect of chlorhexidine, it also reduces Candida albicans adhesion to oral mucosal cells, a factor of importance in the establishment of candidal infection. Clin Infect Dis, 1994 Mar, 18(3), 364 - 8 Use of amphotericin B during pregnancy: case report and review; Dean JL et al.; Unlike the situation with many antimicrobial agents, there is limited experience with the use of amphotericin B during pregnancy . Although reports of fungal infections during pregnancy have been published, few describe fungemia with either Candida or Torulopsis species . We present a case of fungemia due to Torulopsis glabrata that occurred during pregnancy and that was treated with amphotericin B . Drug concentrations were measured in placental tissue, cord serum, and infant serum at delivery . Although the last dose of amphotericin B was administered 4 weeks before delivery, the concentrations in all three specimens were still within the MIC ranges for most strains of Candida albicans and T . glabrata as measured by broth dilution . We speculate that persistent tissue concentrations of amphotericin B most likely contributed to the sustained hypokalemia in the mother and the increased creatinine level in the infant . In the latter case, placental tissue may have served as the reservoir from which amphotericin B was slowly released into fetal circulation. J Bacteriol, 1994 Mar, 176(6), 1702 - 10 A fourth secreted aspartyl proteinase gene (SAP4) and a CARE2 repetitive element are located upstream of the SAP1 gene in Candida albicans; Miyasaki SH et al.; Candida albicans secreted aspartyl proteinases (Sap), products of the SAP genes, which are presumed to act as virulence factors . In the C . albicans strain WO-1, the ability to secrete Sap1 is regulated with switch phenotype, another putative virulence factor . KpnI restriction fragment length polymorphisms differentiate between several distinct SAP1 alleles in laboratory and clinical strains . Both SAP1 alleles from strain WO-1 along with their 5'- and 3'-flanking regions were cloned and sequenced, as were both alleles from another strain, SS . The 5'-flanking regions were remarkably similar in all four of the sequenced alleles over approximately 1,500 nucleotides . S1 analysis revealed that both alleles of WO-1 are transcribed . Characterization of the one allele from strain WO-1 identified a 284-nucleotide insertion flanked by 8-bp direct repeats that shows homology to the CARE2 repetitive element and that is not present in the other alleles . Characterization of the SAP1 alleles also identified a fourth SAP gene (SAP4) that includes an extended leader sequence . SAP4 is positioned upstream, in tandem to SAP1, in all strains tested and may encode another closely related secreted aspartyl proteinase. Mycoses, 1994 Mar-Apr, 37(3-4), 93 - 9 Effects of recombinant human granulocyte-colony stimulating factor on neutropenic mice infected with Candida albicans: acceleration of recovery from neutropenia and potentiation of anti-C . albicans resistance; Hamood M et al.; The treatment of systemic candidal infection in neutropenic patients continues to be a major problem, and only 20% of patients survive despite treatment with amphotericin B (Amph B) . Granulocyte colony-stimulating factor (G-CSF) is a haemopoietic glycoprotein that appears to control the survival, cycle, activation, proliferation and maturation of neutrophil granulocytes and promoter recovery from neutropenia . Confirming previous results, we observed that subcutaneous (s.c.) injection of recombinant human (rh) G-CSF in mice (30 micrograms kg-1 daily) increased the circulating leucocyte count (fourfold) on day 5 of treatment, and led to an expansion of the bone marrow myeloid compartment . The in vivo effect of rhG-CSF on murine resistance to systemic Candida albicans infection was also studied in neutropenic mice . Neutropenia was induced by intraperitoneal injection of a single dose of cyclophosphamide (CPA, 200 mg kg-1) 4 days before C . albicans infection and 2 days before rhG-CSF treatment . rhG-CSF administration showed a protective role on mice infected intravenously (i.v.) with one million C . albicans spores; all the untreated control mice died within 8 days after infection, whereas about 40% of mice treated with rhG-CSF remained alive for the same period . Furthermore, the survival rate was greater in host animals treated with combined Amph B and rhG-CSF than in those treated with Amph B alone . The number of C . albicans colony-forming units (CFU-C . albicans) in the kidney of infected mice was lower in the rhG-CSF-treated group than in the non-treated control mice . This suggests that the severity of infection is decreased in rhG-CSF-treated host animals. J Oral Pathol Med, 1994 Mar, 23(3), 133 - 9 Comparison of a lesion-inducing isolate and a non-lesional isolate of Candida albicans in an immunosuppressed rat model of oral candidiasis; Allen CM et al.; Two distinct strain-related patterns of organism-host interaction on dorsal tongue of immunocompetent rats have been identified for Candida albicans: some isolates induce mucosal lesions, while other isolates penetrate the keratin layer but do not produce a lesion . This study examined the behavior of each of the two types of isolates in a cyclosporin-immunosuppressed rat model . Groups B (normal) and D (cyclosporin) were orally inoculated with a lesion-inducing isolate of C . albicans, while a non-lesional isolate was given to Groups A (normal) and C (cyclosporin) . A typical dorsal tongue lesion developed in 4/18 rats in Group B and in 13/16 in Group D (P = 0.00267) . No significant difference in infection rate between the normal and cyclosporin-treated animals was seen for the non-lesional isolate . The lack of a host inflammatory response associated with the non-lesional isolate may represent an ecologic advantage for the organism. Scott Med J, 1994 Feb, 39(1), 17 - 8 Multiple pancreatic abscesses due to Candida albicans following ERCP; Jalan R et al.; A report is presented of a patient who developed multiple abscesses of the pancreas due to Candida albicans following an Endoscopic retrograde chole-pancreatography (ERCP) for acute pancreatitis . He was not immunocompromised, debilitated and had not had recent surgery . There was complete radiological and clinical resolution of the abscess on prolonged treatment with amphotericin alone . Only a few cases of candidal abscess of the pancreas have been reported, none of them having occurred after an ERCP. Proc Natl Acad Sci U S A, 1994 Feb 1, 91(3), 922 - 6 Candida albicans estrogen-binding protein gene encodes an oxidoreductase that is inhibited by estradiol; Madani ND et al.; Candida albicans, the most common fungal pathogen of humans, possesses an estrogen-binding protein (EBP) that binds mammalian estrogens with high affinity . We report here the cloning and complete nucleotide sequence of a gene encoding a C . albicans EBP . Amino acid sequences obtained from cyanogen bromide fragments of purified EBP were used to design oligonucleotide primers for PCR . An 800-bp product was amplified and used to screen a C . albicans genomic library . A clone was isolated containing an insert with an open reading frame of 1221 nt capable of encoding a protein with 407 amino acids and having a calculated molecular mass of 46,073 Da, the estimated size of EBP . The cloned gene, expressed in Escherichia coli as a lacZ fusion protein, demonstrated high-affinity binding for estradiol and a competition profile comparable to C . albicans wild-type EBP . Northern blots of C . albicans RNA revealed a single transcript of approximately 1600 nt, whereas Southern blots identified three hybridizing fragments . Computer searches of data bases showed that EBP shares a 46% amino acid identity with the old yellow enzyme, an oxidoreductase from Saccharomyces cerevisiae, but was unrelated to the human estrogen receptor as previously speculated . In addition, a 51-amino acid region of EBP is highly conserved among a group of flavoproteins including old yellow enzyme . Expressed EBP was shown to exhibit oxidoreductase activity that could be inhibited by 17 beta-estradiol in vitro . In conclusion, the EBP from C . albicans has no evident homology to the mammalian steroid receptor superfamily but appears to be a member of a recently identified family of flavoproteins. J Leukoc Biol, 1994 Feb, 55(2), 161 - 8 Secretion of TNF-alpha by alveolar macrophages in response to Candida albicans mannan; Garner RE et al.; Resident alveolar macrophages (AM phi) were tested for their ability to respond to Candida albicans mannan . AM phi were found to produce tumor necrosis factor alpha (TNF-alpha) in vitro in response to mannan stimulation . TNF-alpha secretion was measured using ELISA and L929B cellular cytotoxicity assays . Cytotoxicity was neutralized in parallel L929B cell cultures by the addition of rabbit anti-TNF-alpha antibody . Mannan preparations were found to be free of contaminating LPS by Limulus assay . When AM phi were cultivated for 18 h at 37 degrees C, 67 micrograms of mannan stimulated the secretion of approximately 207 U/ml of TNF-alpha . By comparison, AM phi treated with 6.7 micrograms of LPS secreted approximately 257 U/ml of TNF-alpha . Optimal TNF-alpha production occurred between 9 and 18 h after mannan stimulation . Disparate mechanisms for stimulation of TNF-alpha secretion were suggested by differential sugar blockade of LPS- and mannan-induced TNF-alpha secretion . The addition of 2% D-mannose or 2% alpha-methyl-D-mannoside to AM phi cultures blocked mannan- but not LPS-stimulated TNF-alpha secretion . Furthermore, the addition of rabbit anti-mannan antibody to mannan-coated plastic culture dishes prevented TNF-alpha secretion by the mannan-sensitive RAW 264.7 cell line . Moreover, the data suggest that C . albicans mannan stimulated AM phi to secrete TNF-alpha by an LPS-independent receptor mechanism which may also function as a mannose receptor. J Bacteriol, 1994 Feb, 176(3), 756 - 63 Induced chromosome rearrangements and morphologic variation in Candida albicans; Barton RC et al.; We have isolated a mutant of Candida albicans that switches between colony morphologies at high frequencies in a strain with several genetic markers . This strain, 1183, has an altered karyotype with two extra chromosomes . The 1183 karyotype is unstable upon passage . Using DNA transformation with the URA3 gene flanked by sequences from the C . albicans repeat sequence 27A, we have marked individual chromosomes of 1183 and 1161, a related smooth, stable strain . Many transformants contained one or more extra chromosomes, ranging in size from 150 kb to 2.1 Mb . Most were less than 800 kb and appeared to be fragments of a single chromosome . All fragments tested derive from one of the two smallest chromosomes . Six of 13 fragments contained the URA3 gene . In some cases, URA3 was located at the end of a fragment with adjacent telomere repeats . The integrated copy of URA3 was unstable in some 1183 transformants . Our results suggest that 1183 has a mutation affecting genomic stability . A connection between karyotypic changes and morphologic variation has been suggested from studies of several C . albicans strains; however, we find that gross karyotypic and morphological changes are separable processes. Infect Immun, 1994 Feb, 62(2), 742 - 6 Evidence for the presence of a high-affinity laminin receptor-like molecule on the surface of Candida albicans yeast cells; Lopez-Ribot JL et al.; Two polypeptides of 37 and 67 kDa that bind laminin were detected in cell wall extracts of Candida albicans blastoconidia . The 37-kDa species, found only in yeast cell wall extracts, cross-reacted with a rabbit polyclonal antibody (PAb 4160) directed towards the carboxyl-terminal laminin-binding domain present in the human 67-kDa high-affinity laminin receptor (67LR) and its 37-kDa precursor (37LRP), whereas another antibody (PAb 4056), directed against internal domains of 67LR and 37LRP, recognized a 37-kDa species in wall extracts from both blastoconidia and germinated blastoconidia . Indirect immunofluorescence with PAb 4160 showed a patchy binding pattern only on yeast cells that represented about 10% of the entire blastoconidia population. Infect Immun, 1994 Feb, 62(2), 709 - 12 Heterogeneous surface distribution of the fibrinogen-binding protein on Candida albicans; Martinez JP et al.; As detected by indirect immunofluorescence and confocal microscopy, fibrinogen binding was heterogeneously distributed on the surface of Candida albicans . A low level of binding was generally observed homogeneously distributed on some yeast and most hyphal extensions of germ tubes . However, on most hyphal extensions, there were randomly distributed areas of increased expression, as revealed by patches of greater fluorescence intensity. J Gastroenterol, 1994 Feb, 29(1), 1 - 5 Significance of modes of adherence in esophageal Candida albicans; Hoshika K et al.; Although esophageal candidiasis is the most common form of Candida infection in the gastro-intestinal tract, little attention has been directed toward determining the mechanism of its infection . We have already clarified the existence of four modes of adherence of Candida albicans to the esophagus; attachment, subepithelial cell insertion, cavitation, and invasion . This study was undertaken to clarify the significance of each of these modes . Scanning electron microscopic observations were made of esophageal specimens from 8-week-old rabbits infected with Candida albicans IFO 1060 . In this study, attachment and subepithelial cell insertion were found to be the most frequent modes of adherence . Cavitation occurred following subepithelial cell insertion, while invasion occurred following attachment and subepithelial cell insertion . These results suggest that attachment and subepithelial cell insertion play the most important role in the initial stage of adherence . The ratios of these modes for living yeast cells were similar to those for dead yeast cells and beads . This suggests that Candida albicans can gain a foothold on the esophageal epithelium solely by physical contact, after which colonization occurs. Antimicrob Agents Chemother, 1994 Feb, 38(2), 371 - 3 Saperconazole therapy of murine disseminated candidiasis: efficacy and interactions with amphotericin B; Sugar AM et al.; The efficacy of a new triazole antifungal agent, saperconazole, in a murine model of disseminated candidiasis was studied . Mice were intravenously infected with Candida albicans blastoconidia and treated for 14 days with oral saperconazole, intraperitoneal amphotericin B, or a combination of these . Amphotericin B alone was the most efficacious in prolonging survival and in decreasing renal colony counts, usually with complete sterilization of the kidneys by the end of the treatment course . Saperconazole improved survival rates and effected a decrease in renal colony counts, but kidneys were not microbiologically sterilized . Combination therapy with saperconazole and amphotericin B did not result in a decrease in the efficacy of amphotericin B by either end point (survival or renal colony counts) . High-pressure liquid chromatographic analysis of saperconazole concentrations in serum indicated low levels of absorption of the drug . We conclude that saperconazole is effective in the treatment of murine invasive candidiasis and that the theoretical concern about adverse interactions between the two drugs does not apply to the dosages studied in these experiments. Antimicrob Agents Chemother, 1994 Feb, 38(2), 363 - 4 Physicochemical cell damage in relation to lethal amphotericin B action; Beggs WH; A direct relationship between the concentration-dependent rate of amphotericin B-induced K+ release from Candida albicans and the concentration-dependent rate of killing by the drug was established . This relationship together with the observed rapidity of both release and killing action supports the conclusion that the lethal action of amphotericin B is primarily physicochemical in nature. Antimicrob Agents Chemother, 1994 Feb, 38(2), 356 - 9 Aerosolized amphotericin B-liposomes for treatment of systemic Candida infections in mice; Gilbert BE et al.; Mice lethally infected with Candida albicans were exposed to small-particle aerosols containing amphotericin B-liposomes . The drug, when administered twice daily for 2 h (0.58 mg/kg of body weight per day) on days 1, 2, and 3 postinoculation, significantly reduced the numbers of Candida organisms in the kidneys . Aerosol treatment increased the survival time of mice given 2 2-h treatments once a week for 4 weeks . A twice-weekly, 2-h small-particle aerosol administration of amphotericin B-liposomes for 1, 2, or 3 weeks significantly increased both the mean time of survival and percent survival. Antimicrob Agents Chemother, 1994 Feb, 38(2), 223 - 7 Influence of lipoproteins on renal cytotoxicity and antifungal activity of amphotericin B; Wasan KM et al.; We examined the influence of high-density lipoproteins (HDLs) and low-density lipoproteins (LDLs) on the toxicity of amphotericin B (AmpB) to fungal and renal cells . Candida albicans was incubated for 18 h at 37 degrees C with AmpB and deoxycholate (Fungizone) or liposomal AmpB (L-AmpB) (0.1 to 2.0 micrograms of AmpB per ml) in the presence or absence of HDLs or LDLs (0.5 mg of protein per ml) . The MICs of AmpB and L-AmpB, whether or not HDLs or LDLs were present, were similar . LLC PK1 renal cells, derived from primary cultures of pig proximal tubular cells, were incubated for 18 h at 37 degrees C in serum-free medium that contained AmpB and deoxycholate or L-AmpB at 20 micrograms of AmpB per ml, HDLs or LDLs at 0.5 mg of protein per ml, mixtures of AmpB with HDLs or LDLs, and mixtures of L-AmpB with HDLs or LDLs . HDL-associated AmpB was less toxic than AmB to LLC PK1 cells (53.0% +/- 2.5% versus 81.3% +/- 3.6% cytotoxicity; P = 0.01), while LDL-associated AmpB was as toxic as AmpB . L-AmpB, HDL-associated L-AmpB, and LDL-associated L-AmpB were less toxic to LLC PK1 cells than was AmpB (48.3% +/- 1.5%, 25.5% +/- 2.2%, and 52.2% +/- 2.5% versus 81.3% +/- 3.6% cytotoxicity; P = 0.02) . To further understand why HDL-associated AmpB reduced renal cytotoxic effects, the LLC PK1 cells were examined for the presence of HDL and LDL receptors . LLC PK1 cells expressed high-affinity (K(d) = 0.0538 nanograms/ml; 96,000 sites per cell) and low-affinity (K(d) = 222.22 nanograms/ml; 77 sites per cell) LDL receptors but only a low-affinity HDL receptor (K(d) = 71.43 nanograms/ml; 2 sites per cell) . HDL-associated AmpB and LDL-associated AmpB were less toxic than AmpB to trypsinized LLC PK1 cells (46.6% +/- 10.9% and 16.8% +/- 15.98% versus 74.7% +/- 7.7% cytotoxicity; P = 0.02) . HDL-associated AmB and LDL-associated L-AmpB were also less toxic than AmpB to the cells (20.4% +/- 6.2% and 13.5% +/- 8.6% versus 74.7% cytotoxicity; P = 0.01) . The antifungal activities of AmpB and L-AmpB were not altered in the presence of HDLs or LDLs . We conclude that the reduced nephrotoxicity associated with the use of L-AmpB is related to a decreased uptake of AmpB by renal cells when AmpB is associated with HDLs because of the low level of expression of HDL receptors in these cells. J Clin Pharm Ther, 1994 Feb, 19(1), 41 - 6 Microbial contamination and preservative efficacy of topical creams; Na'was T et al.; Nineteen different brands of topical creams were tested in duplicate for microbial growth . The efficacy of their preservation was assessed, when possible, by standard pharmacopoeial procedures . Microbial growth was demonstrated in cultures from all specimens . However, high counts (> 10(4) c.f.u./g) were obtained from eight specimens from six different brands . The microbes were found to belong to different species of bacteria and fungi including Pseudomonas aeruginosa . Two samples of each of the seven, easily emulsifiable creams, were challenged with standard strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans to assess the efficacy of the preservatives used in their preparations . With the exception of a single brand, all creams tested were ineffectively preserved . This study highlights the dangers of marketing domestically prepared topical creams and more effective monitoring of the components for all topical creams is recommended . The inclusion of a proper preservative system in these formulas is essential. J Antimicrob Chemother, 1994 Feb, 33(2), 309 - 18 The effect of fluconazole prophylaxis on fungal colonization in neutropenic cancer patients . Bone Marrow Transplantation Team; Chandrasekar PH et al.; The impact of prophylaxis with 400 mg/day fluconazole on fungal colonization at different body sites was assessed in a randomized, double-blind, placebo controlled study among patients with leukaemia and those undergoing bone marrow transplantation . The study drug was given throughout the period of neutropenia and samples were obtained at weekly intervals . Of the 23 patients in each group, 11 of those given fluconazole and 12 placebo recipients were colonized at entry . The commonest sites were the oropharynx and rectum and Candida albicans was the most frequent isolate . Fluconazole led to a marked reduction in colonization by the second week of treatment to 29% compared with 68% for those given the placebo . Two-weeks after stopping the study regimen there was little change with yeast being isolated from 33% and 81% respectively . Fluconazole was particularly effective in reducing the carriage of C . albicans in the oropharynx from 46% to 0-10% and in maintaining this throughout prophylaxis . Recovery of Candida (Torulopsis) glabrata from the perianal region steadily increased to around 30% in both patient groups and while Candida krusei species were found exclusively in patients given fluconazole, other candida were more common in the placebo group . These results demonstrate that by rapidly reducing the colonization of the alimentary tract, fluconazole eliminates the major reservoir for infection with yeasts other than C . glabrata and C . krusei during the critical period of neutropenia. Microbiology, 1994 Feb, 140 ( Pt 2), 281 - 7 The role of microfilaments and microtubules during pH-regulated morphological transition in Candida albicans; Yokoyama K et al.; Yeast cells of Candida albicans produced germ tubes in a salt-glucose medium containing 4% calf serum at pH 7 and 37 degrees C . Hyphal growth continued for 24 h and the filaments did not revert to yeast cells . When cells were grown at pH 4, reversion to yeast growth was observed, despite the presence of serum . The elongation of hyphae was inhibited within 30 min . The distribution of microtubules and microfilaments during pH-regulated morphological transition was studied by an immunofluorescence technique using an antitubulin antibody with a FITC-conjugated secondary antibody, and by staining with tetramethylrhodaminyl phalloidin for filamentous actin and actin granules . After changing to acidic conditions, microtubules were distributed normally in the cytoplasm; however, microfilaments disappeared from hyphal cells, and actin granules were localized at the site of budding . These results show that microfilaments play an important role during pH-regulated morphological transition. Microbiology, 1994 Feb, 140 ( Pt 2), 271 - 80 The role of the cytoskeleton in the polarized growth of the germ tube in Candida albicans; Akashi T et al.; Cells of the dimorphic yeast Candida albicans are easily induced to germinate in synchrony . Using germinating cells of strain FC18, we examined the effects of several drugs that are known to affect the cytoskeleton on growth and cytoskeletal organization . Cytochalasin A (CA), an inhibitor of actin function, inhibited the germination of the yeast cells and changed the cylindrical expansion of the apex of the germ tube to swelling growth . Effects of CA on the organization of actin were examined with rhodamine-phalloidin (Rh-Ph), which specifically stains F-actin . In CA-untreated cells, Rh-Ph staining resulted in condensed dot-like fluorescence at the growing tip, as well as filamentous fluorescence (actin cables) that ran from the apex to the basal region . In CA-treated cells, condensed dot-like fluorescence was still observed at the swelling tip, but actin cables had disappeared completely . This result indicates that CA does not affect the asymmetrical distribution of actin, and suggests that the actin cables are not required for maintenance of the polarized localization of actin . Benomyl, an anti-microtubule drug, inhibited the germination of yeast cells and the apical growth of germinated cells . Benomyl not only disrupted microtubules (MTs), but also affected the distribution of actin . In benomyl-treated cells, actin dots were randomly dispersed all over the cell . This result indicates that benomyl destroyed the mechanism that maintains the asymmetrical distribution of actin, and suggests that MTs are involved in such a mechanism . The polarized localization of organelles is one of the most important factors associated with dimorphism.(ABSTRACT TRUNCATED AT 250 WORDS) Ann Pharmacother, 1994 Feb, 28(2), 252 - 60 Use of antifungal therapy in hospitalized patients . I . Results prior to the marketing of fluconazole; Grasela TH et al.; OBJECTIVE: To evaluate the use of antifungal agents in hospitalized patients prior to marketing of fluconazole and to assess characteristics associated with their use . DESIGN: A cohort of hospitalized patients receiving topical or systemic antifungal therapy was monitored concurrently . SETTING: Sixty-nine hospitals ranging in size from 100 to more than 500 beds, 70.1 percent affiliated with medical schools . PATIENTS: Participating clinical pharmacists each identified 15 consecutive patients receiving systemic antifungal therapy and 5 consecutive patients receiving topical antifungal therapy at their institutions . Data collection began October 1989 and ended March 1990 . INTERVENTION: All data collected were observational in nature, and no patient intervention was required . MEASURES: Characteristics of patients receiving antifungal therapy were compared using t-tests and chi-square tests . Utilization and patterns of use of antifungal therapy were reported . RESULTS: The most common risk factors necessitating antifungal therapy, in descending order, were: administration of broad-spectrum antibiotics and/or presence of invasive catheters, carcinoma, AIDS, leukemia or lymphoma, diabetes mellitus, solid organ or bone marrow transplantation, and chronic obstructive pulmonary disease . Five hundred seventeen patients received systemic therapy and 464 (89.7 percent) received a single systemic agent . Of these, 242 (52.2 percent) received amphotericin B, 215 (46.3 percent) received ketoconazole, 6 (1.3 percent) received flucytosine, and 1 (0.2 percent) received intravenous miconazole . Fifty-three patients received two systemic agents either concurrently or consecutively . Ketoconazole was most often used for presumed or documented oral, urogenital, or esophageal infections and amphotericin B was the preferred agent for disseminated infections and fungemia (p < 0.001) . Almost half of the patients receiving amphotericin B or ketoconazole (48.3 percent) received these drugs as empiric therapy . Documented infections were more likely to be treated with amphotericin B (54.8 percent) than with ketoconazole (27.4 percent) (p < 0.001) . The predominant fungal isolates were Candida albicans, Candida spp., and unspecified yeasts . Amphotericin B toxicity led to discontinuation of drug therapy in only 5.1 percent of cases . Two hundred sixty-nine patients (34.2 percent) received topical antifungal therapy only . Nystatin oral suspension was prescribed to 65.3 percent of the patients, clotrimazole troches to 23.0 percent, amphotericin B irrigation to 10.9 percent, and nystatin tablets to 0.8 percent . CONCLUSIONS: The utilization patterns of antifungal agents in this survey follow established therapeutic guidelines . Prior to the introduction of fluconazole, amphotericin B was the agent of choice for documented systemic fungal infections . Ketoconazole was more often used for prophylaxis of fungal infections and treatment of oral and esophageal infections. Clin Infect Dis, 1994 Feb, 18(2), 240 - 2 Resistance of Candida albicans to fluconazole during treatment of oropharyngeal candidiasis in a patient with AIDS: documentation by in vitro susceptibility testing and DNA subtype analysis; Redding S et al.; We describe a patient with recurrent episodes of oropharyngeal candidiasis who required progressively higher doses of fluconazole to control and infection . The patient was treated for 14 infections over a 2-year period with doses of fluconazole that ranged from 100 to 800 mg per day . Clinical response, two methods of in vitro susceptibility testing, and molecular epidemiologic techniques were evaluated for 12 of the 14 episodes . Ultimately, the patient became unresponsive clinically to a dose of 800 mg of fluconazole per day . In vitro susceptibility testing of isolates obtained during these successive episodes of infection revealed the development of resistance to fluconazole, and molecular epidemiologic techniques confirmed the persistence of the same Candida albicans strain throughout all 12 episodes. Jpn J Antibiot, 1994 Feb, 47(2), 125 - 8 {Cross-resistance of Candida albicans to several different families of antifungals with ergosterol biosynthesis-inhibiting activity}; Hiratani T et al.; Using 9 strains of Candida albicans mutants resistant to either a triazole-antifungal vibunazole, an imidazole-antifungal ketoconazole or a morpholine-antifungal amorolfine, experiments were conducted to see whether cross-resistance was developed to any of these 3 different families, as well as to an allylamine-antifungal terbinafine . All four compounds are known to selectively inhibit fungal ergosterol biosynthesis . All the mutant strains were found to be more or less resistant to other families of ergosterol biosynthesis inhibitors but not to terbinafine. J Clin Microbiol, 1994 Feb, 32(2), 506 - 9 Comparative evaluation of alternative methods for broth dilution susceptibility testing of fluconazole against Candida albicans; Pfaller MA et al.; A comparative evaluation of methods for broth macro- and microdilution susceptibility testing of fluconazole was conducted with 119 clinical isolates of Candida albicans . Macro- and microdilution testing were performed according to National Committee for Clinical Laboratory Standards recommendations . For reference macrodilution testing, an 80% inhibition endpoint (MIC 80%) was determined after 48 h of incubation in accordance with National Committee for Clinical Laboratory Standards proposed standard M27-P . Microdilution endpoints were scored as the first tube or well in which a prominent reduction in turbidity (score 2 out of a possible 4) was observed compared with the growth control (Micro MIC-2) . Alternative endpoint criteria were assessed independently of the reference MIC 80% and Micro MIC-2 values and included a colorimetric microdilution endpoint determined by using an oxidation-reduction indicator (Alamar Blue; Alamar Bio-sciences Inc., Sacramento, Calif.) . The MICs for the two microdilution test systems were read after 24 and 48 h of incubation . The percentage of fluconazole MICs within 2 doubling dilutions of the macrodilution reference values was 94% for both microdilution tests read at 24 h . Agreement was slightly lower at 48 h and ranged from 91 to 93% . Comparison of Micro MIC-2 and colorimetric microdilution MICs resulted in agreements of 97 and 93% at 24 and 48 h, respectively . These results show excellent agreement among alternative methods for fluconazole susceptibility testing. Am J Med Sci, 1994 Feb, 307(2), 115 - 8 Case report: Candida meningitis with an intradural filling defect 1 year after candidemia; Sarmiento R et al.; Candida albicans meningitis developed in a 55-year-old diabetic female, 1 year after catheter-associated candidemia . It was characterized by a protracted course, lack of meningeal signs, and the presence of an intradural filling defect within the caudal canal . Complete resolution of this filling defect with antifungal therapy implies that it probably represented an inflammatory mass . The development of meningitis after self-limiting candidemia and similar intradural filling defects have not been reported previously. Planta Med, 1994 Feb, 60(1), 41 - 4 Antifungal tests in phytochemical investigations: comparison of bioautographic methods using phytopathogenic and human pathogenic fungi; Rahalison L et al.; The detection limits in two bioautographic assays have been determined for a series of antifungal compounds, including clinically used antimycotics, fungicidal agrochemicals, and various classes of secondary plant metabolites . Target organisms were the filamentous fungus Cladosporium cucumerinum and the yeast Candida albicans . For clinical agents and agrochemicals, the detection limits in the two assays reflected to a certain extent their known spectrum of activity . Most of the plant-derived compounds tested showed a positive response in both assays, but with detection limits varying by a factor up to tenfold . For screening purposes, it is thus advisable to use both tests, as some activities would otherwise go undetected . The MIC values of these substances were determined in order to verify a possible correlation with the detection limit in the bioautographic assays. J Trauma, 1994 Feb, 36(2), 161 - 7 Activation of polymorphonuclear neutrophilic granulocytes following burn injury: alteration of Fc-receptor and complement-receptor expression and of opsonophagocytosis; Vindenes H et al.; Polymorphonuclear neutrophilic leukocytes (PMNLs) play a key role in host defense, and phagocyte dysfunction has been associated with increased susceptibility to infection in patients with thermal injury . We have used flow cytometric analysis (FCM) to longitudinally study PMNL expression of IgG Fc-receptor II (Fc gamma RII) and Fc-receptor III (Fc gamma RIII), as well as the complement receptors CR1 (receptor for C3b) and CR3 (receptor for C3bi) in 22 patients with large burns . Analyses of PMNL complement and immunoglobulin-mediated phagocytosis of Candida albicans were performed in parallel . Burn patient PMNL Fc gamma RIII expression was decreased to 58% of control values at admission, and remained low for the first 3 weeks . The expression of patient PMNL Fc gamma RII was not altered at admission or throughout the hospital stay . The CR1-dependent fluorescence was increased by 62% at admission, and reached a maximum at day 2, 138% greater than that of controls . The CR1 expression then gradually returned to normal at discharge . The PMNL CR3-dependent fluorescence showed an increase of 110% at admission and remained high during the first 3 weeks . The immunoglobulin-mediated phagocytosis was decreased by 12% at admission, whereas the lowest value was observed at day 10, with a reduction of 30% compared with controls . The patient PMNL complement-mediated phagocytosis of C . albicans was increased by about 160% at admission, and reached a maximum at day 2, before it gradually decreased to control levels at discharge . The expression of complement receptors correlated positively, whereas the expression of Fc gamma RIII correlated negatively, with total body surface area (TBSA) burn.(ABSTRACT TRUNCATED AT 250 WORDS) Transplantation, 1994 Feb, 57(3), 398 - 402 Candida carriage in the alimentary tract of liver transplant candidates; Kusne S et al.; Thirty randomly selected patients with advanced chronic liver disease, which had been evaluated for possible liver transplantation, were sampled endoscopically at 7 alimentary tract locations to assess the frequency and amount of Candida carriage . Eighty-one percent (127/156) of the samples obtained contained Candida and 53% (82/156) yielded high counts (> 300 CFU/ml) . The most predominant Candida species isolated at each site was Candida albicans, which accounted for 103 (64%) of the 160 fungal isolates . The other Candida species isolated included C tropicalis 30 (19%), C krusei 16 (10%), and C glabrata 11 (7%) . Although the number of sites at which yeast was present and the quantities of yeast at each site varied widely among the patients studied, 100% of the patients had Candida in at least one site of the gastrointestinal tract . Eighty-six percent (24/28) of the duodenal aspirates contained Candida and 50% (14/28) of the duodenal samples contained greater than 300 CFU/ml . A positive culture from the stomach was a reliable predictor of the presence of Candida in the duodenum (P = 0.0001), but a positive culture at no other site readily predicted the presence of Candida at yet another site . Importantly, there was no correlation between the presence or absence of Candida in either oral or rectal swabs and colonization at other anatomic sites within the gastrointestinal tract . These findings are important in liver transplantation, particularly in those cases in which the bowel has been opened to create a choledochojejunostomy anastomosis . The operative attempts to reduce gastrointestinal fungal carriage using oral antifungal agents may be justified before liver transplantation in an effort to lower the risk of posttransplantation fungal infections, particularly in those patients expected to have a Roux-en-Y choledochojejunostomy biliary reconstruction. J Infect Dis, 1994 Feb, 169(2), 452 - 6 A subset of proteins found in culture supernatants of Candida albicans includes the abundant, immunodominant, glycolytic enzyme enolase; Sundstrom P et al.; Immunoblot analysis showed that enolase is one of a subset of proteins found in cell supernatants of Candida albicans . Enzyme assays on whole cell extracts indicated that enolase is an abundant protein, comprising 0.7% and 2.0% of the total protein from yeast and hyphal forms of C . albicans, respectively . Comparison of enolase enzyme activities in whole cell extracts and cell culture supernatants showed the enzyme to be located primarily within cells . Extracellular glyceraldehyde-3-phosphate dehydrogenase activity was absent or lower than that of enolase, despite equivalent intracellular levels . The results suggest that enolase, released from fungi in the absence of host factors, may contribute to enolase found circulating in the blood of patients with hematogenously disseminated candidiasis . In addition, the release from cells of highly immunogenic fungal proteins, such as enolase, may be important in defining the selective stimulation of host antifungal responses during infection. J Infect Dis, 1994 Feb, 169(2), 369 - 74 Antifungal activity of recombinant human macrophage colony-stimulating factor in models of acute and chronic candidiasis in the rat; Vitt CR et al.; Models of acute and chronic candidiasis were developed in Fischer 344 rats to evaluate the therapeutic efficacy of recombinant human macrophage colony-stimulating factor (rhM-CSF) alone and in combination with the antifungal agent fluconazole . In the acute model, rats were challenged by intravenous injection with 2 x 10(6) Candida albicans, approximately 4 times the LD50 . Daily subcutaneous (sc) bolus injections of rhM-CSF for 10 days plus a single sc bolus dose of 0.3 mg/kg of fluconazole improved the median survival time from 5 days (32% survival) with fluconazole alone to > 30 days (88% survival) in the rhM-CSF- and fluconazole-treated rats . In the chronic model, daily sc bolus injections of rhM-CSF for 10 days plus a single sc bolus dose of 1.0 mg/kg of fluconazole decreased the median titer of C . albicans cultured from the kidneys by 10-fold at 15 and 30 days after infection . These studies showed that rhM-CSF treatment improved the therapeutic outcome in both the acute and chronic rat model of candidiasis when used with fluconazole, a standard fungistatic agent. J Chemother, 1994 Feb, 6(1), 50 - 2 Antibiotics affecting gastrointestinal colonization of mice by yeasts; Samonis G et al.; A group of three-month old, male Crl:CD1(ICR) BR mice, was fed chow containing Candida albicans, while another group of the same type of mice was fed regular chow . Both groups were treated subsequently with either antibiotics or normal saline for 10 days . Stool cultures were performed before treatment, at the end of treatment, and one week after the end of treatment, to determine the level of colonization of the gastrointestinal tract by the yeast . The stools of mice fed Candida and treated with antibiotics had substantially higher Candida counts than control mice fed C . albicans and treated with saline . The highest concentrations of the yeast were observed in the stools of mice treated with cefotaxime as compared to those of mice treated with pefloxacin, amikacin and amoxicillin . No Candida was found in the stools of mice fed regular chow and treated with antibiotics or saline . Dissemination of Candida was not observed in the visceral organs of any mouse. Comp Immunol Microbiol Infect Dis, 1994 Feb, 17(1), 77 - 84 Decline in the phagocytic function of alveolar macrophages from mice exposed to cigarette smoke; Ortega E et al.; An investigation was made of the effects of a short and acute exposure to cigarette smoke on the capacity of alveolar macrophages from mice to carry out all stages of the phagocytic process . Cigarettes were commercial 80 mm filter cigarettes that contained 17 mg of tar and 1.1 mg of nicotine per cigarette . The acute exposure of each animal was with one cigarette for 15 min (until the complete consumption of the cigarette) in a box-shaped plastic chamber, 7732 cm3 vol and 450 cm2 floor surface, with a 3 cm dia airhole . Animals were sacrificed immediately after the exposure to the smoke . The results showed no differences either in the adherence or in the chemotaxis capacities between alveolar macrophages from control mice and from mice exposed to cigarette smoke . However, there was a significant decline both in attachment capacity and in ingestion capacity for Candida albicans . The reason for this was a decline in the number of macrophages with phagocytic capacity (percent of phagocytosis) and in the number of C . albicans phagocytized per cell (phagocytic efficiency) . The conclusion is that a short, acute exposure to a smoke-filled atmosphere induced a decrease in the phagocytic function of alveolar macrophages. Bull Tokyo Dent Coll, 1994 Feb, 35(1), 23 - 6 Efficacy of Listerine antiseptic against MRSA, Candida albicans and HIV; Yamanaka A et al.; The antiseptic activity of Listerine and Cool Mint Listerine against methicillin-resistant Staphylococcus aureus (MRSA), Candida albicans, human immuno deficiency virus (HIV) and oral bacteria was examined in this study . Exposure for 30 seconds to Listerine killed MRSA completely . Exposure for 30 seconds significantly decreased viable cells of C . albicans . More than 60% of HIV was inactivated by a 30 second exposure to 50% Listerine . Listerine exhibited a potent bactericidal effect against cariogenic and periodontopathic bacteria . Cool Mint Listerine had almost the same antiseptic effect against tested microorganisms . Listerine appears to be effective for killing etiologic microorganisms of opportunistic infection in the oral cavity. J Invest Dermatol, 1994 Feb, 102(2), 197 - 204 Prevention of ultraviolet radiation-induced suppression of contact and delayed hypersensitivity by Aloe barbadensis gel extract; Strickland FM et al.; We investigated the ability of Aloe barbadensis gel extract to prevent suppression of contact hypersensitivity (CHS) and delayed-type hypersensitivity (DTH) responses in mice by ultraviolet (UV) irradiation . Local immune suppression was induced in C3H mice by exposure to four daily doses of 400 J/m2 UV-B (280-320 nm) radiation from FS40 sunlamps, followed by sensitization with 0.5% fluorescein isothiocyanate (FITC) through the irradiated skin . Topical application of 0.167-1.67% Aloe gel after each irradiation significantly reduced this suppression . Aloe treatment partially preserved the number and morphology of Langerhans and Thy-1+ dendritic epidermal cells in skin, compared to those in the skin of mice given only UVR or UVR plus the vehicle . Experiments using a single (2 kJ/m2) dose of UVR followed by Aloe treatment showed that the effect of Aloe was not due to screening of the UVR . Systemic suppression of DTH to Candida albicans or CHS to FITC was induced in C3H mice exposed to 5 or 10 kJ/m2 UV-B radiation, respectively, on shaved dorsal skin and sensitized 3 d later with a subcutaneous injection of formalin-fixed Candida or FITC painted on unirradiated, ventral skin . Treatment of the UV-irradiated skin with Aloe immediately after irradiation prevented suppression of both DTH to Candida and CHS to FITC . Aloe treatment did not prevent the formation of cyclobutyl pyrimidine dimers in the DNA of UV-irradiated skin or accelerate the repair of these lesions . These studies demonstrate that topical application of Aloe barbadensis gel extract to the skin of UV-irradiated mice ameliorates UV-induced immune suppression by a mechanism that does not involve DNA damage or repair. Infect Immun, 1994 Feb, 62(2), 738 - 41 Lack of effect of Candida albicans mannan on development of protective immune responses in experimental murine candidiasis; Garner RE et al.; Candida albicans mannoprotein (MAN) administered to mice before or during immunization with viable C . albicans downregulates MAN-specific delayed hypersensitivity . In the experiments reported here we determined the effect of MAN downregulation on protective immunity in minimally immunized mice, i.e., mice exposed to C . albicans either intradermally or intragastrically, and in maximally immunized mice, i.e., mice immunized by a combination of intradermal and intragastric exposure, in experimental systemic candidiasis . MAN suppression did not induce statistically significant alterations in the protective responses in experimental candidiasis, although 8 of 12 groups of mice treated with MAN had fewer CFU of C . albicans in their kidneys than their non-MAN-treated counterparts . The results emphasize the lack of correlation of delayed hypersensitivity with protection in candidiasis and suggest that MAN may contain epitopes involved in the protective response. Infect Immun, 1994 Feb, 62(2), 509 - 19 Modulation of cell surface-associated mannoprotein antigen expression in experimental candidal vaginitis; De Bernardis F et al.; The monoclonal antibody (MAb) AF1 recognizes an oligosaccharide epitope present on highly immunogenic and immunomodulatory mannoproteins (MP) of Candida albicans . The expression of this epitope (AF1-MP) during experimental candidal vaginitis was studied in two strains of C . albicans (3153 and CA-2) which were equally vaginopathic but differed in the mode of hypha formation in the vagina . In both strains, immunofluorescence of vaginal samples, taken 1 h after challenge, revealed an intense, MAb AF1-specific labelling of the yeast cells . This labelling was very scarce in fungal cells taken at 24 h and on subsequent days during the development of filamentous forms . Electron-microscopic gold immunolabelling observations showed that molecules carrying AF1-MP spanned the entire cell wall in the initial yeast cells but were absent on the cell surface and in the outermost, capsular layer of the cell wall of the germ tubes and filamentous forms . In both strains, at any time and for any form of intravaginal growth, AF1-MP was clearly expressed in the cytoplasm and cytoplasmic vesicles, and was fully incorporated into the inner layers of the cell wall . As seen by immunofluorescence, the vaginal fluid from C . albicans-infected rats did not hinder the expression of AF1-MP on the yeast cells surface in vitro . In electron-microscopic gold immunolabelling, a hypha-specific MAb (3D9) labelled the surface of the hyphal but not of the yeast cells of C . albicans harvested from rat vagina . Overall, these data strongly suggest that cell surface expression of MP antigen is modulated during intravaginal growth and morphogenesis of C . albicans. Eur J Immunol, 1994 Feb, 24(2), 440 - 4 Interleukin-2 prevention of apoptosis in human neutrophils; Pericle F et al.; Evidence is presented that interleukin (IL)-2 maintains viability of human polymorphonuclear cells (PMN) in culture by preventing these cells from undergoing programmed cell death (PCD) and induces the synthesis of new RNA and protein . Our laboratory has recently discovered that human PMN constitutively express IL-2 beta receptor and more importantly, PMN are able to respond functionally to IL-2 by enhanced growth inhibitory activity against an opportunistic fungal pathogen, Candida albicans . We now report that IL-2 was able to interfere with the PCD process and reduce the number of apoptotic PMN to < 40% in 72-h culture . Freshly isolated PMN usually underwent a time-dependent aging process and > 80% of PMN cultured in medium alone for 72 h showed morphologic features of PCD as depicted by hematoxylin and eosin staining as well as by electron microscopy . During the PCD process, untreated PMN not only exhibited condensed nuclear structure and decrease in cell size, but also displayed DNA fragmentation . DNA fragmentation in PMN was prevented by IL-2 . Prevention of PCD by IL-2 was associated with an increase in new RNA and protein synthesis in PMN, which may reflect cytokine induction, such as tumor necrosis factor, as we have recently shown . Thus, our data expands our current understanding of PMN in that they may be an active component of the immune system, with a longer life-span when activated than expected. Brain Res, 1994 Jan 28, 635(1-2), 1 - 8 Fever of unknown origin: due to C . albicans or other fungi acting on the hypothalamus? Barwick VS, Wooten MH, Bradfield JF, Myers RD. Recently, it was shown that cerebrospinal fluid (CSF) contaminated with the fungus Trichosporon beigelii produces an intense fever when the organism is microinjected directly into the thermosensitive region of the anterior hypothalamic preoptic area (AH/POA) . The purpose of this study was to determine if the AH/POA possesses a corresponding sensitivity to another fungal organism, Candida albicans . In adult male Sprague-Dawley rats, an intracerebral cannula was implanted stereotaxically above the AH/POA and a radio transmitter for the continuous recording of body temperature (Tb) was placed in the peritoneal cavity . After recovery, one of two solutions was microinjected in the AH/POA: a pyrogen-free, filtered artificial CSF and a second cultured with C . albicans in a concentration of approximately 12 x 10(8) organisms/ml . Whereas the filtered CSF failed to evoke a significant rise in Tb, C . albicans produced a febrile response of 0.8-1.5 degrees C in the rats within 1 h after its microinjection into the AH/POA . This fever persisted typically for > or = 12 h but after 24 h Tb returned to the baseline . Histological examination of the cerebral tissue postmortem revealed focally extensive granulomatous encephalitis with disseminated inflammation throughout the parenchyma of rats given repeated microinjections of C . albicans . Since C . albicans is a highly potent pyrogen acting directly on thermosensitive neurons, it is envisaged that a massive accumulation of the organism within the brain could be responsible pathologically for the protracted fever "of unknown origin" which gives rise to clinical morbidity. J Biol Chem, 1994 Jan 14, 269(2), 802 - 4 Active site mapping of affinity-labeled rat oxidosqualene cyclase; Abe I et al.; Rat liver oxidosqualene cyclase (OSC), a 78-kDa membrane-bound enzyme, was purified and labeled with the mechanism-based irreversible inhibitor, {3H}29-methylidene-2,3-oxidosqualene (Abe, I., Bai, M., Xiao, X.-Y., and Prestwich, G . D . (1992) Biochem . Biophys . Res . Commun . 187, 32-38) . A 6-kDa CNBr peptide was separated by Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted to a polyvinylidene difluoride membrane . The sequence of the first 30 amino acids of this peptide were determined by Edman degradation and showed unexpectedly high similarity to the fungal OSC from Candida albicans (50% identity with Arg413-Val442) and to the bacterial squalene cyclase from Alicyclobacillus (formerly Bacillus) acidocaldarius (37% identity with Lys356-Leu385) . Further, radioanalysis clearly established that the two adjacent Asp residues in the highly conserved region (Asp-Asp-Thr-Ala-Glu-Ala or DDTAEA) were equally labeled by the irreversible inhibitor . This result provides the first information on the structural details of the active site of OSC and shows for the first time the ancient lineage of this vertebrate enzyme to ancestral eukaryotic and prokaryotic cyclases . Interestingly, the covalently modified DDXX(D/E) sequence of rat liver OSC showed surprising similarity to the putative allylic diphosphate binding site sequence of sesquiterpene cyclases and prenyl transferases. Dtsch Med Wochenschr, 1994 Jan 7, 119(1-2), 13 - 8 {Unusual course of candidiasis of the central nervous system}; Lisch S et al.; Six months after an attack of pyelonephritis, adnexitis and candida colpitis an 18-year-old girl developed some clouding of consciousness . On neurological examination she showed organic behavioural changes, discrete anisocoria and possible meningism . Computed tomography revealed hydrocephalus and signs of increased cerebrospinal fluid (CSF) pressure . CSF contained 2336/3 cells, while total protein was raised to 7.0 g/l and lactate concentration to 6.85 mmol/l . Glucose concentration in CSF was 51 mg/dl and 75 mg/dl in serum . As tuberculous meningitis was suspected, treatment was started with four tuberculostatic drugs, but there was no improvement . Five weeks later microscopic CSF examination showed fungal spores and nonbranching hyphae . The maximal candida haemagglutination titre in CSF was 1:2048 . CSF culture grew Candida albicans . The further course was complicated by side effects to the antimycotic drugs (amphotericin B between 4.5 and 45 mg daily; flucytosine 1.7 g four times daily) and recurrent obstruction in the ventricular system requiring repeated neurosurgical interventions . However, full cure was achieved after seven months' hospital treatment. Am J Surg, 1994 Jan, 167(1), 145 - 50 Total parenteral nutrition, bacterial translocation, and host immune function; Shou J et al.; Total parenteral nutrition (TPN) is associated with increased infectious complications in trauma and perioperative patients compared with enteral nutrition support . This study evaluated the effects of TPN on splenocyte and peritoneal macrophage (PM phi) function and intestinal bacterial translocation . Male Wistar rats underwent central vein cannulation and were randomized to isocaloric feeding of a regular chow diet (RD) plus saline infusion or TPN for 7 days . Splenocytes and PM phi were harvested to assess concanavalin A mitogenesis, superoxide production, and Candida albicans phagocytosis . Bacteria-positive mesenteric lymph nodes (MLNs) were found in 77% (10 of 13) of TPN-fed rats compared with 17% (2 of 12) of RD-fed rats (p < 0.05) . Splenocyte mitogenesis, PM phi superoxide production, and C . albicans phagocytosis were significantly decreased in the TPN group compared with results in the RD group . In a second study, rats received RD, TPN, and parenteral nutrition (PN) with 10% or 20% of calories given as oral chow (PN and 10% chow and PN and 20% chow) for 7 days . PN and 10% chow reversed the TPN-induced suppression of C . albicans phagocytosis . PN + 20% chow significantly increased splenocyte mitogenesis, PM phi superoxide production, and C . albicans phagocytosis and killing to normal levels and was associated with a decreased incidence of bacteria-positive MLN . Thus, administration of TPN is associated with impaired PM phi microbicidal and splenocyte proliferative function . These defective cellular functions were reversed with a small amount of oral feeding. J Allergy Clin Immunol, 1994 Jan, 93(1 Pt 1), 93 - 9 The production of interferon-gamma in response to a major peanut allergy, Ara h II correlates with serum levels of IgE anti-Ara h II; Dorion BJ et al.; The current study was undertaken to examine the potential role of T cells in the pathogenesis of peanut allergy . Peripheral blood mononuclear cells (PBMCs) from patients with peanut allergy, patients with asthma, and nonatopic normal control subjects were assessed for proliferation after stimulation with a 17 kd major peanut allergen (Ara h II), ovalbumin, casein, soy, and Candida albicans . We found that Ara h II and C . albicans induced significantly higher levels of proliferation than ovalbumin, casein, and soy . Because interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) play critical roles in IgE regulation, we assessed the production of these cytokines after stimulation with C . albicans and Ara h II . C . albicans stimulated similar levels of IFN-gamma in all three study groups . In contrast, after stimulation with Ara h II, culture supernatants from PBMCs of subjects with peanut allergy contained significantly lower levels of IFN-gamma than did the PBMCs of the two control groups (p = 0.02) . More important, there was a significant (p = 0.05) inverse correlation between the serum IgE anti-Ara h II levels and IFN-gamma production by PBMCs from the respective peanut-allergic patients . IL-4 protein was not detected in culture supernatants of PBMCs stimulated with Ara h II . However, amplification of cytokine gene transcripts by polymerase chain reaction did demonstrate IL-4 expression in Ara h II-stimulated PBMCs from both patients with peanut allergy and control subjects . These data suggest that the level of IFN-gamma production in response to Ara h II may be an important factor in determining the development of peanut-specific IgE responses. Cell Immunol, 1994 Jan, 153(1), 239 - 47 Heterogeneous secretory response of phagocytes from different anatomical districts to the dimorphic fungus Candida albicans; Blasi E et al.; In the present study, we have examined the ability of phagocytes from different anatomical districts to discriminate between the two morphogenetic forms of Candida albicans . We have demonstrated that resident peritoneal macrophages (RP-M phi) and thioglycollate-elicited peritoneal macrophages (TP-M phi) were able to distinguish between the hyphal (H-Candida) and the yeast (Y-Candida) form of the fungus, since TNF production was observed only upon exposure of RP-M phi and TP-M phi to H-Candida . In contrast, splenic macrophages (S-M phi), bone marrow-derived macrophages (BM-M phi) and polymorphonuclear neutrophils (PMN) did not discriminate between the two forms because S-M phi and PMN produced TNF regardless of the morphogenetic status of the fungus, while BM-M phi did not . Under the same experimental conditions, we failed to observe IL-1 production from any of the phagocytic cell populations examined, with the exception of PMN . This implies that the interaction between phagocytes and C . albicans triggers differential secretory responses depending upon the morphogenetic status of the fungus and the anatomical localization of the immune cells. J Nutr, 1994 Jan, 124(1 Suppl), 144S - 148S Dietary nucleotides: cellular immune, intestinal and hepatic system effects; Carver JD; Investigations with animals demonstrate that dietary nucleotides influence immune function . Restriction of dietary nucleotides in mice decreases several indices of cell-mediated immunity as well as resistance to challenge with Staphylococcus aureus or Candida albicans . Spleen cells of mice maintained on nucleotide-free diet produce less interleukin-2 and have lower natural killer cell cytotoxicity and macrophage activation than those of animals fed nucleotide-supplemented diets . In vivo lymphoproliferative response, macrophage phagocytic activity and expression of interleukin-2 receptor and lyt1 surface marker are also lower in animals fed nucleotide-free diets . At 2 mo of age, infants fed breast milk or nucleotide-supplemented infant formula exhibit increased natural killer cell activity compared with infants fed unsupplemented formula . Dietary nucleotide restriction in animals may also result in hepatic lipid accumulation and decreased mucosal height and gut wall thickness . Adenosine monophosphate, a mediator of hepatic and small bowel blood flow, may play a unique role among the nucleotides studied . In conclusion, de novo synthesis and salvage of nucleotides is a metabolically costly process . An exogenous source of nucleotides from the diet may optimize the function of rapidly dividing tissues, particularly when growth is rapid and the diet is low in nucleotides. Arch Surg, 1994 Jan, 129(1), 99 - 105 Exudative neutrophils . Modulation of microbicidal function in the inflammatory microenvironment; Yee J et al.; OBJECTIVE: To determine whether the inflammatory microenvironment primes neutrophils for increased microbicidal activity . DESIGN: In vitro studies of host defense were performed on surgical patients . SETTING: A tertiary care, university hospital . PATIENTS: A volunteer sample of hospitalized preoperative, noninfected surgical patients . INTERVENTION: Exudative neutrophils were collected from skin-blister chambers and functionally compared with circulating neutrophils . METHODS: Flow cytometry was used to evaluate directly neutrophil microbicidal activity (using fluoresceinated Candida albicans), formyl-methionyl-leucyl-phenylalanine (fMLP)-induced superoxide production (using 123 dihydrorhodamine), and surface expression of CD11b, CD16, and the fMLP receptor . In vitro tumor necrosis factor alpha was used to determine the possibility and extent of further priming in both circulating and exudative neutrophils . RESULTS: Exudative polymorphonuclear neutrophils have enhanced microbicidal activity, superoxide production, and expression of CD11b, CD16, and the fMLP receptor . Exogenous tumor necrosis factor was able to prime circulating neutrophils but did not further augment superoxide production in exudative neutrophils . CONCLUSION: The microbicidal activity of neutrophils is enhanced after exudation and is associated with neutrophil priming . The inability of exogenous tumor necrosis factor to further augment superoxide production after exudation suggests that this priming has been maximized. Chest, 1994 Jan, 105(1), 317 - 8 Role of Candida albicans in chronic hypersensitivity pneumonitis; Ando M et al.; An autopsy was performed on a patient who had chronic hypersensitivity pneumonitis that was observed for 9 years . The patient was a farmer who developed symptoms every March through July during the use of moist hay that was infected heavily with Candida . Precipitins and an inhalation challenge test to C albicans were positive . We interpret the role of C albicans in this case. J Med Vet Mycol, 1994, 32(1), 59 - 64 Relationship between Candida albicans epidermolytic proteinase activity and virulence in mice; Louie A et al.; Five strains of Candida albicans with previously characterized epidermolytic acid proteinase activity were evaluated for virulence following intravenous (i.v.) injection in mice . Increased proteinase activity was associated with increased virulence in female, NYLAR mice receiving 10(6) cells i.v . Mean mortality times (1.25, 2.0, 2.0, 4.25 and 19.6 days, in groups of 20 mice for each of the five strains) correlated directly with degree of proteinase activity . Three of the strains were selected for additional in vivo study and the association between increased proteinase activity and increased mortality rates was confirmed in dose-response studies in two additional strains of mice . The mean survival times appeared to be independent of fungal growth rate in vitro . These results support the positive correlation between proteinase activity and virulence. J Med Vet Mycol, 1994, 32(1), 21 - 30 Arrays of Candida albicans pseudohyphae that protect the organisms from neutrophil fungicidal mechanisms in experimental infections of mice; Sohnle PG et al.; Experimental subcutaneous Candida albicans infections in mice were used to examine the manner in which this pathogen is cleared in animals recovering from cyclophosphamide-induced leucopenia . In this system, infections at the inoculation sites progressed rapidly during a 6 day period of leucopenia to form arrays of parallel filamentous organisms that effectively isolated those in the interior from contact by neutrophils, even when the leucopenia had resolved . Dense collections of organisms also developed at sites of metastatic infection in the kidneys . A majority of the organisms were found to be viable when they were retrieved from the infected subcutaneous sites of animals that had recovered from leucopenia and whose abscesses had begun to drain spontaneously . Removal of the protective arrays of fungal cells appeared to be accomplished by drainage of abscess contents through the surface of the skin or into the collecting system of the kidney . Drainage of the subcutaneous abscesses did not occur in the cyclophosphamide-treated animals until after the neutrophilic infiltrates had developed, suggesting that this drainage process was mediated by neutrophils rather than by the organisms themselves . In summary, the above findings demonstrate that C . albicans infections in leucopenic hosts may progress to the extent that they would be very difficult to clear solely through the microbicidal processes of returning neutrophils . However, neutrophils also appear to promote the removal of masses of viable fungal cells to the exterior of the body. J Med Vet Mycol, 1994, 32(1), 13 - 20 Purification and characterization of fatty acid synthase from Candida albicans strain 4918 and two derived spontaneous cerulenin-resistant mutants; McElhaney-Feser GE et al.; Fatty acid synthase from three strains of Candida albicans (parental strain 4918, and two spontaneous cerulenin-resistant mutants, 4918-2 and 4918-10) has been purified and characterized . In all three cases the purification protocol included ammonium sulfate precipitation, fractionation with butyl-Toyopearl, differential centrifugation and sedimentation velocity centrifugation . Inclusion of protease inhibitors, aprotinin, leupeptin and pepstatin was a prerequisite to maximize recoveries . Polyacrylamide gel electrophoresis analysis demonstrated protocol efficacy and showed the apparent molecular mass of the two enzyme sub-units from each strain to be 195 kDa and 210 kDa . The Km (malonyl-CoA) and Vmax of each fatty acid synthase were similar . In contrast, inactivation kinetics of the respective enzymes in the presence of cerulenin showed enzyme activity from both mutants to differ significantly from the parent and from each other . Other experiments suggested in vivo cerulenin resistance of mutant strains is not solely attributable to enzyme alteration. J Med Vet Mycol, 1994, 32(1), 1 - 11 Specific inhibition of acid proteinase secretion in Candida albicans by Lys-Nva-FMDP; Milewski S et al.; Secretion of aspartic (acid) proteinase by Candida albicans is inhibited by the action of a new anticandidal agent, L-lysyl-L-norvalyl-{N3-(4-methoxyfumaroyl)}-L-2,3-diamino pro panoic acid (Lys-Nva-FMDP), at low, even sub-minimum inhibitory concentrations . The observed phenomenon is a direct consequence of inhibition of the enzyme, glucosamine-6-phosphate synthase . As a result of this inhibition, biosynthesis of candidal mannoproteins is markedly reduced . A possible correlation between general inhibition of mannoprotein biosynthesis and acid proteinase secretion is suggested . The reported inhibition of acid proteinase secretion by Lys-Nva-FMDP is more specific than the previously described effects of methyl patricin, 5-fluorocytosine and fenticonazole. Allergy, 1994 Jan, 49(1), 50 - 6 Identification of allergen components of the opportunistic yeast Pityrosporum orbiculare by monoclonal antibodies; Zargari A et al.; The yeast Pityrosporum orbiculare (P . orbiculare) is a member of the normal human cutaneous flora, but it is also associated with several clinical manifestations of the skin . We have previously observed IgE-binding components in P . orbiculare extracts, using sera from patients with atopic dermatitis . In the present study, we raised several monoclonal antibodies (MoAbs) against P . orbiculare to characterize some of its antigens, and used Candida albicans (C . albicans) as a control . We obtained several IgG1 MoAbs which specifically recognized P . orbiculare in ELISA . Two of these were selected for immunoblotting studies on P . orbiculare, and two patterns of reactivity emerged . Firstly, one MoAb showed a distinct band at a molecular mass of 67 kDa . In the second pattern, a sharp band at about 37 kDa appeared . In contrast, the IgM antibodies raised reacted with a 14-kDa component; but they reacted with C . albicans in addition to P . orbiculare . The IgG1 antibodies seemed to react with proteins, as their ability to react in ELISA with extract pretreated with protease was greatly reduced . In contrast, IgM MoAbs were much less affected, suggesting that they recognized nonprotein components . To determine whether these MoAbs-binding components were also recognized by human IgE, we adopted a radioimmunoassay (RIA) using the MoAbs as catcher antibodies . Both the 67-kDa and the 37-kDa components were IgE-binding proteins . P . orbiculare RAST positive sera were scored as positive in the RIA, whereas the control serum was not. Farmaco, 1994 Jan, 49(1), 51 - 5 Antifungal agents . 7 . Dichlorophenylpyrrylimidazolylmethane derivatives: synthesis and antifungal activities; Massa S et al.; The synthesis and antifungal activities of some dichlorophenyl-1H-pyrrol-2-yl-1H-imidazol-1-ylmethane derivatives substituted at pyrrole nitrogen are reported . When tested against Candida albicans and Candida spp., some derivatives were found to be from two to four times less active than miconazole, bifonazole and ketoconazole, used as standard controls. Zh Mikrobiol Epidemiol Immunobiol, 1994 Jan-Feb, (1), 14 - 6 {Macrophage interaction with Candida albicans in immunodepression}; Pakhomova EN et al.; The interaction of fungi of the species C . albicans with macrophages has been studied in vivo under the conditions of immunosuppression resulting from the action of immunosuppressive agents, belonging to different groups with respect to the chemical nature and mechanism of their action (cyclophosphamide, azathioprine, hydrocortisone acetate, methotrexate, cyclosporin A) . The data obtained in this study indicate that under the conditions of immunity disturbances macrophages have decreased functional activity . Azathioprine and cyclosporin A have the greatest inhibiting influence on macrophages during their interaction with fungi, while cyclophosphamide has the least influence. Microbiology, 1994 Jan, 140 ( Pt 1), 167 - 71 Candida albicans aspartic proteinase cleaves and inactivates human epidermal cysteine proteinase inhibitor, cystatin A; Tsushima H et al.; It is known that the cysteine proteinase inhibitor, cystatin, has a defence function against exogenous pathogens . Human epidermal cysteine proteinase inhibitor, cystatin A, which is a member of the cystatin family, is localized in the upper epidermal layer . In this study, the relationship between cystatin A and Candida aspartic proteinase (CAP), a putative Candida virulence factor, was studied . CAP activity was not affected by human epidermal cystatin A, while 90% of cystatin A activity was lost after incubation with CAP for 12 h at 37 degrees C . Human epidermal cystatin A was cleaved into small peptides by CAP, and the released peptides had no cystatin activity . These results suggest that CAP may induce an imbalance between cysteine proteinase and its inhibitor in cutaneous Candida infectious lesions through the degradation and inactivation of epidermal cystatin A. J Nat Prod, 1994 Jan, 57(1), 79 - 83 Discobahamins A and B, new peptides from the Bahamian deep water marine sponge Discodermia sp; Gunasekera SP et al.; Discobahamin A {1} and discobahamin B {2} are two bioactive peptides isolated from a new species of the Bahamian deep water marine sponge Discodermia . The discobahamins are inhibitors of the growth of Candida albicans, and the isolation and structure elucidation of 1 and 2 by nmr and chemical methods is described. Gynecol Obstet Invest, 1994, 37(2), 110 - 4 Detection of anti-Candida albicans IgE antibodies in vaginal washes from patients with acute vulvovaginal candidiasis; Regulez P et al.; Vaginal washes from 55 women were investigated by means of an ELISA method for the presence of IgE antibodies against Candida albicans . These antibodies were detected in 87.1% of patients with clinical acute vulvovaginal candidiasis (group I), 100% of patients with suspected vulvovaginal candidiasis but negative by microscopy and culture (group II), 0% of asymptomatic carriers (group III) and 33.3% of uninfected controls (group IV) . Statistically significant differences were observed comparing groups I and II vs . groups III and IV . The highest IgE vaginal antibody titers were mostly at the expense of serotype A C . albicans strains, which represented 83.3% of the C . albicans isolates . Non-C . albicans species also showed very low IgE levels . No correlation between serum and vaginal IgE was found . Furthermore, a second determination of vaginal IgE levels was performed in 3 patients . A decrease in IgE levels concomitant to a decline in clinical symptoms was observed in all of them after treatment. Can J Microbiol, 1994 Jan, 40(1), 77 - 81 Adherence of Candida albicans germ tubes to murine tissues in an ex vivo assay; Lopez-Ribot JL et al.; Adhesion of Candida albicans germ tubes to murine tissues was examined . An ex vivo assay previously employed to examine adhesion of yeast cells of C . albicans was adapted for use with germ tubes . Binding of germ tubes to kidney, liver, spleen, and lymph node tissues was found to occur throughout the tissue section, with little morphologic specificity . In general, more organisms adhered to spleen and lymph node tissues than to kidney and liver tissues . Observation of adhesion with scanning organisms adhered to spleen and lymph node tissues than to kidney and liver tissues . Observation of adhesion with scanning electron microscopy showed three germ tube-tissue interactions described as loose, tight, or embedded. J Appl Bacteriol, 1994 Jan, 76(1), 68 - 74 The use of impedance for preservative efficacy testing of pharmaceuticals and cosmetic products; Connolly P et al.; Impedance was investigated for its applicability to preservative efficacy testing of pharmaceuticals and cosmetics . A good correlation between impedance detection time (Td) and total colony counts (colony-forming units (cfu)) was obtained for untreated suspensions of Staphylococcus aureus, Candida albicans, Aspergillus niger and Pseudomonas aeruginosa in phosphate-buffered saline (PBS) . A good correlation between Td and the number of cfu was also obtained for suspensions of test organisms treated for varying contact periods with selected concentrations of chlorhexidine, methyl paraben and phenoxyethanol in PBS, and methyl paraben in cetomacrogol cream, but these correlations were significantly different from those for untreated suspensions . It was found that for any given number of cfu the Td for preservative treated cells was extended . It is concluded that impedance represents a valid method for preservative efficacy testing of pharmaceuticals and cosmetics which could be used to achieve more comprehensive but economic screening of formulations against a wider range of preservative systems and concentrations than is the current approach where only a limited range of systems are tested because of the workload involved. Antimicrob Agents Chemother, 1994 Jan, 38(1), 90 - 5 In vivo pharmacokinetics and pharmacodynamics of topical ketoconazole and miconazole in human stratum corneum; Pershing LK et al.; A direct study evaluating whether differential drug uptake of topical 2% miconazole and 2% ketoconazole from cream formulations into human stratum corneum correlated with differential pharmacological activity against Candida albicans was investigated in healthy human subjects . A single 24-h topical dose of 2% ketoconazole cream or 2% miconazole cream was applied unoccluded, at the same dose (2.6 mg of formulation per cm2 of surface area), at four skin sites on both ventral forearms of six human subjects . At the end of the treatment, residual drug was removed with a tissue from all sites and the treated site was tape stripped 11 times, either 1, 4, 8, or 24 h later . The first tape disc was discarded . The remaining tape discs, 2 through 11, were combined and extracted for drug quantification by high-performance liquid chromatography and bioactivity against C . albicans growth in vitro . Topical 2% ketoconazole produced 14-, 10-, and 7-fold greater drug concentrations in stratum corneum than 2% miconazole at 1, 4, and 8 h after a single topical dose . Ketoconazole and miconazole concentrations in the stratum corneum were similar 24 h after drug removal . Tape disc extracts from 2% ketoconazole-treated skin sites demonstrated significantly greater bioactivity in the bioassay than 2% miconazole . The increased efficacy of 2% ketoconazole compared with that of 2% miconazole in vitro reflects their differential uptake into the stratum corneum and inherent pharmacological activity . Tape stripping the drug-treated site in conjunction with a bioassay is therefore a useful approach in the determination of bioavailability of topical antifungal agents. Antimicrob Agents Chemother, 1994 Jan, 38(1), 45 - 8 Improved medium for fluconazole susceptibility testing of Candida albicans; Rodriguez-Tudela JL et al.; We have compared fluconazole susceptibilities of 92 clinical isolates of Candida albicans by broth microdilution in two different media: standard RPMI 1640 (RPMI) and the same medium supplemented with 18 g of glucose per liter (RPMI-glucose) . Preparation of media, drugs, and inocula, as well as incubation conditions, followed the preliminary recommendations of the National Committee for Clinical Laboratory Standards (Villanova, Pa.) antifungal agent working group for broth macrodilution tests with antifungal agents, adapted to microdilution . Microtiter plates were agitated for 5 min before spectrophotometric readings were performed with an automatic plate reader set at 405 mm . The MIC endpoint was defined as an inhibitory concentration calculated from the turbidimetric data as a function of the turbidity in the drug-free control wells . The mean absorbances in the drug-free wells in RPMI and RPMI-glucose were, respectively, 0.38 (41.6% transmission) and 0.99 (10.2% transmission) (P < 0.001; Student's t test) . Despite the increased growth in RPMI-glucose, 98.9% of the C . albicans strains tested for fluconazole susceptibility yielded similar MICs (+/- 1 dilution) in both media . Moreover, strains with decreased susceptibility to fluconazole displaying similar MICs in both media are easier to detect in RPMI-glucose because of the greater differences between turbidimetric readings in wells with grown or fluconazole-inhibited cultures . This objective turbidimetric method, with an easy-to-read improved medium (RPMI with glucose), together with previous experience of the National Committee for Clinical Laboratory Standards antifungal agent subcommittee, could overcome some of the present problems associated with lack of reproducibility of azole susceptibility testing. Antimicrob Agents Chemother, 1994 Jan, 38(1), 13 - 22 Novel aspect of amphotericin B action: accumulation in human monocytes potentiates killing of phagocytosed Candida albicans; Martin E et al.; The influence of low doses of amphotericin B on the capacity of human monocytes to kill Candida albicans was investigated . Killing rates were quantified by a novel flow cytometric assay and were found to be 37% +/- 3% (standard error of the mean) after 3 h . Preincubation of monocytes for 6 to 20 h with low concentrations of amphotericin B (0.2 microgram/ml) resulted in a markedly augmented fungicidal capacity . Enhancement of killing was 80% +/- 11% (standard error of the mean) over that by the controls . This effect did not appear to be due to amphotericin B-dependent monocyte activation; the respiratory burst and expression of human leukocyte antigen-DR were unaltered, and no stimulation of interleukin-1 beta release occurred . Cell-associated amphotericin B was extracted with acetonitrile and was quantified by scanning spectrophotometry . Amphotericin B appeared to accumulate in the cells, and intracellular concentrations attained after overnight incubation in 1 microgram of the drug per ml were estimated to be in the range of 50 fg per cell . The fact that intracellular accumulation was responsible for the enhanced fungicidal capacity of monocytes was supported by the findings that killing of Staphylococcus aureus remained normal and enhancement of killing of an amphotericin B-resistant C . albicans strain was minimal . Dramatic enhancement of monocyte fungicidal capacity probably extends to other amphotericin B-susceptible fungi and could represent a hitherto unrecognized determinant underlying the curative properties and prophylactic efficacy of this drug. J Clin Microbiol, 1994 Jan, 32(1), 228 - 31 Detection of Candida albicans and other yeasts in blood by PCR; Holmes AR et al.; Primers complementary to the region of genes coding for rRNA in Candida albicans were used in PCRs to detect yeast DNA extracted from blood samples containing various Candida species . One fragment (105 bp) was amplified from all yeasts tested, whereas a second (684 bp) was only amplified when C . albicans DNA was present . The level of sensitivity was 15 +/- 5 (mean +/- standard error) CFU of C . albicans per ml of blood. J Clin Microbiol, 1994 Jan, 32(1), 217 - 9 Simplified adsorption method for detection of antibodies to Candida albicans germ tubes; Ponton J et al.; Two modifications that simplify and shorten a method for adsorption of the antibodies against the antigens expressed on both blastospore and germ tube cell wall surfaces (methods 2 and 3) were compared with the original method of adsorption (method 1) to detect anti-Candida albicans germ tube antibodies in 154 serum specimens . Adsorption of the sera by both modified methods resulted in titers very similar to those obtained by the original method . Only 5.2% of serum specimens tested by method 2 and 5.8% of serum specimens tested by method 3 presented greater than one dilution discrepancies in the titers with respect to the titer observed by method 1 . When a test based on method 2 was evaluated with sera from patients with invasive candidiasis, the best discriminatory results (sensitivity, 84.6%; specificity, 87.9%; positive predictive value, 75.9%; negative predictive value, 92.7%; efficiency, 86.9%) were obtained when a titer of > or = 1:160 was considered positive. FEMS Microbiol Lett, 1994 Jan 1, 115(1), 77 - 82 Analysis of hybrids of Candida albicans formed by protoplast fusion; Law C et al.; A number of hybrids obtained by fusion of protoplasts of complementary auxotrophic strains of Candida albicans were analysed for growth rates, cell volume, DNA content, stability and adherence to human buccal epithelial cells . SDS-PAGE analysis of the cell wall proteins indicated that the hybrid cell walls contained many more of the proteins associated with one parent than the other . The adherence values of the hybrids were closest to the parent with which they shared most cell wall proteins and it is suggested that the hybrids contain the genome of this parent along with one or more of the chromosomes of the other. Chem Pharm Bull (Tokyo), 1994 Jan, 42(1), 85 - 94 Optically active antifungal azoles . III . Synthesis and antifungal activity of sulfide and sulfonamide derivatives of (2R,3R)-2-(2,4-difluorophenyl)-3-mercapto-1-(1H-1,2,4-triazol-1-yl)-2-b utanol; Tasaka A et al.; In an effort to find potent antifungal agents, optically active sulfur-containing triazole derivatives, sulfides (3) and sulfonamides (4), were prepared and evaluated for antifungal activity against Candida albicans in vitro and in vivo . The sulfides (3) were prepared by the reaction of (2R,3R)-2-(2,4-difluorophenyl)-3-mercapto-1-(1H-1,2,4-triazol-1-yl )-2-butanol (1) with various heteroarylmethyl chlorides in the presence of sodium methoxide . The sulfonamides (4) were synthesized starting from the disulfide (15) in three steps including oxidation of the corresponding sulfenamides (17) . Some of the sulfur-containing triazole derivatives (3, 4) showed strong protective effects against candidosis in mice. Clin Microbiol Rev, 1994 Jan, 7(1), 29 - 42 Adhesins and ligands involved in the interaction of Candida spp . with epithelial and endothelial surfaces; Hostetter MK; Adhesion of candidal species to the epithelium of the gastrointestinal or genitourinary tract stands as a critical first step in the pathogenesis of candidal infection . After colonization and replication at mucosal surfaces, Candida albicans and other pathogenic species may penetrate the mucosal barrier, enter the vascular tree, and disseminate hematogenously . The consequences of this pathogenic cascade evoke considerable morbidity and mortality, especially among immunocompromised patients . Thus, interactions of C . albicans and other candidal species with epithelium and endothelium may lead to serious consequences for the human host . This review evaluates candidate candidal adhesions for epithelial and endothelial surfaces, with emphasis on the specificity of the interaction, the inhibitors that have been employed, and the ligands that have been identified on mammalian cells or matrices . Three types of interactions are described: protein-protein interactions, lectin-like interactions, and incompletely defined interactions in which the adhesive ligand is as yet unidentified . Special attention is given to the roles of integrin-like proteins . Differences in the mechanisms of candidal attachment to epithelium and endothelium are delineated . Last, on the basis of the available literature, avenues of potentially fruitful investigation are proposed. Arch Microbiol, 1994, 162(1-2), 33 - 40 Characterization of a major cell wall antigen and potential adhesin in three strains of Candida albicans; Jenq W et al.; Selected strains of Candida albicans were examined to reveal the surface antigenicity and biochemical nature of major cell wall proteins that also were shown to serve as cellular adhesins on human buccal epithelial cells . Confirmation of the adhesive properties of these cells was made by scanning electron microscopy and immunofluorescence microscopy . Particular attention was directed at the clinical isolate KM-302 . By means of indirect immunofluorescence staining, the KM-302 blastoconidia absorbed rabbit anti-C . albicans ATCC-32354 serum, revealing specific localization of surface antigens on germ tubes and pseudohyphae . Extracellular polymeric material and the cell wall extract of C . albicans KM-302 blastoconidia were found to contain a major surface antigen of 49 kDa that exhibited 42% adhesion inhibition in vitro . Of considerable significance is that immunogold localization by electron microscopy showed the antigen to be almost exclusively cell wall bound . This major antigen, identified in affinity and gel filtration chromatography fractions, was composed of 4% carbohydrate and 95.7% protein and had an isoelectric point of 6.1 . The major antigen also showed a high level of similarity with that of C . albicans strain SC-5314 inasmuch as the major antigen of that strain had carbohydrate and protein compositions of 4 and 95.5%, respectively . Both of these strains also possessed the same percent of adhesion inhibition on human buccal epithelial cells. Am J Nephrol, 1994, 14(2), 113 - 20 Fungal peritonitis in patients on peritoneal dialysis; Michel C et al.; Fungal peritonitis (FP) is a serious complication of peritoneal dialysis, both in terms of morbidity and mortality . Available data on the effectiveness of fluconazole in eradicating FP without catheter removal are still controversial . We reviewed 20 FP cases that occurred among 325 patients who underwent peritoneal dialysis in our center between January 1984 and January 1992, in order to establish whether a profile of patients at risk of developing FP could be identified and to evaluate the effectiveness of fluconazole in treating FP (7 cases) . Age, sex, a particular cause of end-stage renal disease, and the presence of diabetes did not correlate significantly with the development of FP . The risk of FP increased in patients on immunosuppressive treatment . Sixteen of our 20 patients had bacterial peritonitis during the month before they developed FP . Nineteen were treated with antibiotics . Neither the type of bacterial organism isolated during the bacterial peritonitis preceding FP nor modality and duration of antibiotic treatment correlated significantly with the development of FP . Patients who subsequently developed FP were more frequently treated with antibiotics while in hospital (p < 0.001) . Candida species accounted for 15 of our 20 FP cases (75%), with Candida albicans being by far the most common isolate . Treatment strategies varied among the 20 patients . The combination of intravenous or intraperitoneal administration of 5-fluorocytosine and oral administration of fluconazole was used in 7 cases: only 1 patient was cured without catheter removal, 1 patient died within the first 4 days of treatment, removal of peritoneal catheter was necessary in the other 5 patients.(ABSTRACT TRUNCATED AT 250 WORDS) Microbios, 1994, 79(318), 27 - 30 Quantitative determination of Candida albicans in sea water; Mates A; Candida albicans counts were assessed in Mediterranean sea water using MCA media . From 116 seawater samples C . albicans was detected in 22 samples of 100 ml sea water . In sixteen samples only one colony, and in six samples two colonies, of C . albicans were found and the fungus does not appear to be an important contaminant of sea water. J Med Vet Mycol, 1994, 32(2), 133 - 40 Electrophoretic karyotypes of isolates of Candida albicans from hospitalized patients; Doi M et al.; Electrophoretic karyotypes, namely, the patterns of chromosome-sized DNAs that have been separated by pulsed-field gel electrophoresis, have been reported to vary among different strains of Candida albicans, and can be useful for the delineation of strains . A previous study showed that almost all the electrophoretic karyotypes of isolates of C . albicans collected from the vagina of outpatients were distinguishable from one another . A few of the isolates had a common atypical karyotype and these isolates all had the same phenotype . This report describes the electrophoretic karyotype analysis of isolates from various specimens taken from hospitalized patients . Karyotypes were analysed with respect to the origins of the isolates in terms of two parameters: the number of bands resolved on the gel and a code of four numbers which corresponded to the number of bands that could be separated in each of four sets of chromosome sizes . No specific karyotype was correlated with either the nature of the specimen or the hospital ward to which the patient had been admitted . Most isolates gave a pattern similar to that of FC18 and no atypical karyotypes were found. J Med Vet Mycol, 1994, 32(2), 123 - 32 Biochemical and pharmacological factors causing induction and suppression of germination of Trichosporon beigelii; Walsh TJ et al.; Trichosporon beigelii is an emerging fungal pathogen, which is morphologically characterized by blastoconidia, arthroconidia and hyphae . The non-hyphal forms of T . beigelii germinate to form hyphae in plasma in vitro and in tissues in vivo, suggesting possible pathophysiological significance of this process . Little is known, however, about the mechanisms of germination of T . beigelii . We therefore studied relevant biochemical and pharmacological factors that may regulate germination of T . beigelii . Germination was significantly enhanced by temperature at 37 degrees C, chemically defined cell culture media such as RPMI-1640, plasma, physiological pH,