Bidirectional stimulation of the white-opaque transition of Candida albicans by ultraviolet irradiation

Most strains of Candida albicans are capable of switching spontaneously and at high frequency between a number of phenotypes distinguishable by colony morphology. The switching frequency of Candida albicans strain WO-1 between two predominant phenotypes, 'white' and 'opaque', and a minor phenotype, 'fuzzy', increased dramatically with low doses of ultraviolet irradiation that killed less than 20% of the population. The ultraviolet irradiation effect continued to be expressed over many generations as evidenced by stimulated sectoring. Ultraviolet irradiation stimulated switching in both the white-to-opaque and opaque-to-white direction, suggesting that a common mechanism functions in both directions.     

Eburicol, lichesterol, ergosterol, and obtusifoliol from polyene antibiotic-resistant mutants of Candida albicans.

Two classes of polyene-resistant mutants were isolated from survivors of N-methyl-N'-nitro-N-nitrosoguanidine treatment of a wild-type Candida albicans. An analysis of the major sterols of one class revealed an accumulation of lichesterol and fecosterol while the other class accumulated eburicol, obtusifoliol, and lanosterol with minor quantities of C28 sterols.     

Differentiation of pathogenic species of Candida by their recovery characteristics following ultraviolet irradiation

Each of seven pathogenic species of Candida exhibits a unique pattern of light and dark recovery responses to ultraviolet irradiation. C. guilliermondii, C. parapsilosis and C. pseudotropicalis photoreactivate whereas C. albicans, C. krusei, C. stellatoidea and C. tropicalis do not. Within eachof these groups, individual species are distinguishable by whether or not they express differential dark recovery during postirradiation growth at 25 C or 37 C on oxidative vs fermentative carbon sources, on inorganic vs amino acid nitrogen sources or in the presence rather than absence of ergosterol. Equivalent recovery patterns are obtained for species of Candida and the ascosporogenous species which are their corresponding perfect forms. These observations indicate strongly that the postirradation recovery is a reliable, species-specific characteristic of yeasts.These studies were added in part by a contract (AT11-1)-1772 with the U.S. Atomic Energy Commission.     

The effect of altered ergosterol content on the transport of various amino acids in Candida albicans.

Candida albicans cells have low levels of ergosterol when grown in ascorbic acid-supplemented media. When cells are grown in hydroquinone-supplemented media, the ergosterol levels became higher as compared to normal cells. The uptake of lysine, glycine, glutamic acid, proline, methionine and serine is reduced in hydroquinone-supplemented cells. In contrast to hydroquinone-supplemented cells, the rate and level of accumulation of these amino acids are higher in ascorbic acid-supplemented cells. Nystatin-resistant isolates of C. albicans with low ergosterol contents also exhibit an increased rate and level of accumulation of these amino acids. The uptake of phenylalanine and leucine remained unaffected by such a change in ergosterol levels brought about by different supplementation of the media. The results demonstrate a correlation between ergosterol levels and amino acids uptake. Contrary to various reports, the rate of K+ efflux does not seem to correlate with the amino acid uptake in C. albicans cells.     

Emerging role of lipids of Candida albicans, a pathogenic dimorphic yeast.

It is clear that C. albicans lipids have gained tremendous importance in recent years. In addition to being a barrier for entrance of various metabolites, it also provides the site of action for the synthesis of enzyme(s) involved in cell wall morphogenesis and antifungal action. While alterations in lipid composition during a yeast to mycelia transition have been observed, in most of the studies, lipid fluctuations reported could have been due to various environmental factors involved in the induction of morphogenesis [4,5]. A clear understanding of lipid biosynthesis and metabolic blocks due to antifungal action is likely to shed further light on selective interactions of antifungals. Despite the multifacet role of lipids in various functions of this pathogenic yeast, their exact involvement is poorly understood. The situation is little better with regard to ergosterol and its metabolism. Ergosterol is, indeed, important for anti-candidal activity and appears to be involved in the morphogenesis of C. albicans. The fluctuation in phospholipid composition have led to altered properties of plasma membrane namely, membrane fluidity, transport activities and drug sensitivity, which suggest that-a critical level of individual phospholipid is important for proper functioning of the plasma membrane. What the exact role is of individual phospholipid is far from clear. Many unanswered questions relating to the role of PI and sphingomyelin in signal transduction, involvement of phospholipases in the maintenance of phospholipid composition, and role of lipid transfer proteins in assembly and asymmetry of lipids are some aspects which merit further work.     

Effect of supplementation of ergosterol on miconazole action in vivo and in vitro in Candida albicans.

Ergosterol was observed to alter the lipid composition of C. albicans 3153 selectively, resulting in steep rise in ergosterol content with marginal changes in other lipids content. Supplementation of ergosterol in presence or absence of cerulenin made cells more protective towards the miconazole drug. However, when the same experiments were carried out with liposomes prepared from the lipid extracts of control and supplemented cells, the pattern obtained show a little deviation from the in vivo experiments thus indicating that in addition to lipids, other cell components like proteins also affect interaction of miconazole with C. albicans.     

氨基酸对白念珠菌形态学影响的研究

目的初步探讨单个氨基酸对白念珠菌形态学的影响。方法用0．67％的酵母氮源基础培养基和2％葡萄糖配制成SD合成培养基,37％恒温摇床培养,研究单个天然氨基酸对白念珠菌形态学的影响,并分别通过不添加碳源和厌氧条件下培养观察对精氨酸诱导的菌丝的影响。结果在含10mmol／L的L－精氨酸的SD液体培养基中,可见大量的菌丝。在含10mmol／L的L一半胱氨酸、L．苏氨酸、L－缬氨酸和L－色氨酸的sD液体培养基中,可见典型的酵母细胞,未见菌丝。在含10mmol／L的其他单个氨基酸的SD液体培养基中可见混合的酵母和菌丝结构。在不含氨基酸或含各种天然氨基酸的SD固体培养基上,白念珠菌的菌落均光滑。但在含10mmol／L的L-精氨酸固体培养基上,光滑的菌落周围可见小的突起,镜下可见菌丝。无氧条件下,无论有无碳源,含精氨酸的SD培养液中白念珠菌只能形成酵母细胞,生长部分受到抑制。结论精氨酸可以诱导白念珠菌菌丝形成,厌氧条件下精氨酸不能诱导白念珠菌菌丝形成。     

Effects of human lactoferrin on the cytoplasmic membrane of Candida albicans cells related with its candidacidal activity

Human lactoferrin is an innate host defence protein with antimicrobial activity that exerts a candidacidal effect in a cation concentration-dependent manner. We investigated the ability of this cationic protein (with an isoelectric point of 8.7) to permeabilize the cytoplasmic membrane of Candida albicans cells. Despite minor K(+)-release in lactoferrin-treated C. albicans cells, the killing effect was not related to an extensive membrane permeabilization, as indicated by: (a) the non-release of macromolecular cytosolic constituents; (b) the non-permeabilization for extracellular propidium iodide nor for intracellular accumulated calcein; and (c) the inability to disrupt the phospholipid bilayer of 8-aminonaphthalene-1,3,6, trisulfonic acid/p-xylene-bis-pyridiniumbromide-loaded liposomes. These results suggest that lactoferrin exerts its candidacidal effect through a mechanism different from membrane permeabilization described for other cationic peptides.     

Effects of irradiation on fatty acid structure of membrane lipids of the sarcoplasmic reticulum]

It has been shown that single local X-irradiation (0.21 C/kg) of the rabbit hind limb in the early period of acute radiation injury (1 and 14 h) causes a decrease in saturation of sarcoplasmic reticulum membrane lipids. It is mainly connected with a decreased saturation of the total fraction of phosphatidyl serine, phosphatidyl inositol and phosphatidyl choline. The above changes can increase permeability of the sarcoplasmic reticulum membranes for Ca(2+)-ion after X-irradiation.     

Biochemical effects of miconazole on fungi. II. Inhibition of ergosterol biosynthesis in Candida albicans.

The effects of the antifungal agent miconazole nitrate on the ergosterol biosynthesis in Candida albicans were investigated after in vitro contact with the drug for 1, 4, 16 and 24 h. A time- and dose-(2.10^?10–10^?4 M) dependent inhibition of [¹⁴C]acetate incorporation into ergosterol was observed. Fifty percent inhibition of the acetate incorporation into ergosterol was found after 1 h incubation in the presence of 10^?9 M miconazole. Simultaneously 24-methylenedihydrolanosterol, lanosterol, obtusifoliol, 4,14-dimethylzymosterol and 14-methylfecosterol accumulated.The accumulation of 14 α-methyl sterols suggests that this antifungal agent is a potent inhibitor of one of the metabolic steps involved in the demethylation at C-14. The absence of 24-methyl sterols and of sterols with a C-22 [23] double bond in miconazole treated C. albicans indicates that miconazole also inteferes with the reduction of the 24(28)-double bond and with the introduction of the 22(23)-double bond.Miconazole also intervenes to a small extent in triglyceride synthesis. However, in all circumstances studied, ergosterol biosynthesis was affected at lower doses than those interfering with the acetate incorporation into triglycerides. 16 and 24 h of incubation in the presence of miconazole (≥ 10^?6 M) also resulted in an increased fatty acid synthesis.It is suggested that the miconazole-induced inhibition of the C-14 demethylation may be at the origin of the previously observed permeability changes in miconazole treated C. albicans.     

Simple rapid identification of Candida albicans with emphasis on the differentiation between Candida albicans and Candida stellatoidea

Summary Subcultures ofC. albicans, made from Sabouraud agar, grown at room temperature for 48 hours, were inoculated into a 10 times saline dilution of Sabouraud liquid medium and left in the incubator for 45–60 minutes at 37° C, transferred to corn meal agar plates and incubated at 37° C for 18–24 hours.Small portions of the surface agar containing the yeasts from these plates were pressed under cover glasses and examined under the oil immersion lens.Under these conditions,C. albicans cultures were observed to produce only yeast-like cells, whereasC. stellatoidea cultures contained predominantly abundant, long, thin mycelia.     

Effects of ultraviolet B irradiation on cutaneous leishmaniasis

Protection against many infectious diseases is mediated by cellular immunity in the competent host. Ultraviolet (UV) radiation, a component of sunlight, is a potent suppressor of cell-mediated immune responses. Suzanne Holmes Giannini discusses the possible relevance of ambient levels of UVB to pathogenesis and immunity in infectious diseases, with special reference to cutaneous leishmaniasis.     

Rapid purification of secretory acid proteinase from Candida albicans and its characterization.

Secretory acid proteinase from C. albicans was purified from culture supernatant to apparent homogeneity by ion-exchange chromatography. Two isozymes of the proteinase were resolved using a novel chromatofocusing method. The enzyme, which appears to be a glycoprotein, consists of a single polypeptide chain with glutamine at the N-terminus. Its molecular weight is about 45,000 and isoelectric point is pH 4.6. At pH 5, the proteinase is stable at 45 degrees C for at least 15 min. It has a broad substrate specificity. With BSA as a substrate, Km was determined to be 1.6 x 10(-4) M. The enzyme is inhibited by pepstatin and thus is a carboxyl proteinase. It undergoes autocatalytic digestion at or below pH 5.0. The kinetics of induction of proteinase by various proteins are also reported.