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Serological differences between the multiple amine oxidases of yeasts and comparison of the specificities of the purified enzymes from Candida utilis and Pichia pastoris.

1. Antiserum to purified methylamine oxidase of Candida boidinii formed precipitin lines (with spurs) in double-diffusion tests with crude extracts of methylamine-grown cells of the following yeast species: Candida nagoyaensis, Candida nemodendra, Hansenula minuta, Hansenula polymorpha and Pichia pinus. No cross-reaction was observed with extracts of Candida lipolytica, Candida steatolytica, Candida tropicalis, Candida utilis, Pichia pastoris, Sporobolomyces albo-rubescens, Sporopachydermia cereana or Trigonopsis variabilis. Quantitative enzyme assays enabled the relative titre of antiserum against the various methylamine oxidases to be determined. 2. The amine oxidases from two non-cross-reacting species, C. utilis and P. pastoris, were purified to near homogeneity. 3. The methylamine oxidases, despite their serological non-similarity, showed very similar catalytic properties to methylamine oxidase from C. boidinii. Their heat-stability, pH optima, molecular weights, substrate specificities and sensitivity to inhibitors are reported. 4. The benzylamine oxidases of C. utilis and P. pastoris both oxidized putrescine, and the latter enzyme failed to show any cross-reaction with antibody to C. boidinii methylamine oxidase. Benzylamine oxidase from C. boidinii itself also did not cross-react with antibody to methylamine oxidase. The heat-stability, molecular weights, substrate specificities and sensitivity to inhibitors of the benzylamine/putrescine oxidases are reported. 5. The benzylamine/putrescine oxidase of C. utilis differed only slightly from that of C. boidinii. 6. Benzylamine/putrescine oxidase from P. pastoris differed from the Candida enzymes in heat-stability, subunit molecular weight and substrate specificity. In particular it catalysed the oxidation of the primary amino groups of spermine, spermidine, lysine, ornithine and 1,2-diaminoethane, which are not substrates for either of the Candida benzylamine oxidases that have been purified. 7. Spermine and spermidine were oxidized at both primary amino groups; in the case of spermidine this is a different specificity from that of plasma amine oxidase. 8. Under appropriate conditions, P. pastoris benzylamine/putrescine oxidase (which is very easy to purify) can be a useful analytical tool in measuring polyamines.

Candida↗

Insulin-dependent diabetes mellitus and oral soft tissue pathologies: II. Prevalence and characteristics of Candida and Candidal lesions.

OBJECTIVE: To assess the prevalence of Candida albicans and oral infection with Candida in patients with insulin-dependent diabetes mellitus (IDDM). STUDY DESIGN: This cross-sectional study compared the prevalence of candidiasis in 405 subjects with IDDM and 268 nondiabetic control subjects. Assessments included evidence of clinical manifestations of candidiasis and a quantitative measure of Candida pseudohyphae in a cytologic smear from the midline posterior dorsal tongue. RESULTS: More subjects with IDDM than control subjects without IDDM (15.1% vs 3.0%) were found to have clinical manifestations of candidiasis, including median rhomboid glossitis, denture stomatitis, and angular cheilitis. IDDM subjects were also more likely to have any Candida pseudohyphae in their cytologic smears (23.0% vs 5.7%; P <.0001), as well as pseudohyphae counts of >10/cm(2) (7.1% vs 0.8%; P <.0001). Diabetic subjects with median rhomboid glossitis were more likely to have a longer duration of IDDM and complications of nephropathy and retinopathy. Denture stomatitis was associated with smoking, retinopathy, higher Candida counts, poor glycemic control, and longer duration of IDDM. A multivariate regression analysis found 3 factors to be significantly associated with the presence of Candida pseudohyphae in the subjects with IDDM: current use of cigarettes (odds ratio, 2:4), use of dentures (odds ratio, 2:3), and elevated levels of glycosylated hemoglobin (odds ratio, 1:9). The use of antimicrobials, immunosuppressants, or drugs with xerostomic side effects was not related to the presence of Candida. CONCLUSIONS: Candida pseudohyphae and oral soft tissue manifestations of candidiasis were more prevalent in subjects with IDDM than in control subjects without diabetes. The presence of Candida pseudohyphae was significantly associated with cigarette smoking, use of dentures, and poor glycemic control.

Adult↗

Isolation of Candida species from the oral cavity and fingertips of complete denture wearers.

STATEMENT OF PROBLEM: Candida species usually colonize in the oral cavity of denture wearers and may also colonize on their fingers because of frequent manual manipulation of the dentures. PURPOSE: This study investigated the association between oral and fingertip candidal isolation in a group of denture wearers. MATERIAL AND METHODS: Oral rinse and fingerprints obtained from 25 healthy male complete denture wearers were microbiologically investigated for candidal growth, and isolated Candida species were identified with a germ tube test and a commercially available yeast identification system. Denture cleanliness, hand washing, and denture wearing and handling habits were recorded for each subject. RESULTS: Candida species were isolated from the oral cavity of 15 (60%) and fingertips of 11 (44%) subjects. Ten (66.7%) subjects had concomitant oral and fingertip candidal isolation, whereas 5 (33.3%) subjects had only oral Candida (P=.005). CONCLUSION: The hands of denture wearers who had oral Candida were significantly more colonized with Candida species than oral Candida-free subjects. Further studies identifying Candida species to the strain level and the significance of hand Candida as a source for mouth reinfection are needed.

Adult↗

Characterization and purification of the chorionic gonadotropin-like protein binding site in Candida albicans.

Recent studies from our laboratory have shown that human chorionic gonadotropin (hCG), human luteinizing hormone (hLH), and CG-like proteins from Xanthomonas maltophilia (xCG) and Candida albicans (CaCGLP) induce Candida albicans transition from blastospores to hyphal forms. Xanthomonas maltophilia CG-like protein (xCG), hCG, and hLH also bind to Candida albicans blastospores with a high-affinity nM Kd, indicating that these substances can control Candida albicans pathogenicity. The work reported herein describes the purification of the binding site for these CG-like proteins from Candida albicans. The purification developed involved alcohol extraction followed by affinity chromatography. The product obtained was a protein of 64-69 kDa, as analyzed by sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE), Western blot and Sephadex G-100 column chromatography. This binding site reacted in a Western blot with both 125I-hCG and 125I-CaCGLP. Purified CaCGLP was able to displace specifically 125I-hCG bound to Candida albicans blastospores. Scatchard plot analysis showed that the Kd of this reaction was of high affinity in the nM range, and also indicated the presence of one single binding site. These results lead us to conclude: 1) Candida albicans possesses a binding site which is able to bind hCG, hLH and CG-like proteins from Xanthomonas maltophilia and Candida albicans; 2) This binding site is a hydrophobic protein of approximately 64 kDa and; 3) We postulate that CaCGLP is the natural ligand for this binding site and this system is used to control Candida albicans transition and, therefore, pathogenicity.

Binding Sites↗

Comparison of a new commercial colorimetric microdilution method with a standard method for in-vitro susceptibility testing of Candida spp. and Cryptococcus neoformans.

The Sensititre Yeast One method (AccuMed International Ltd, East Grinstead, UK) is a microplate-based procedure that incorporates an oxidation-reduction indicator, Alamar Blue, for the in-vitro testing of five antifungal agents (amphotericin B, fluconazole, itraconazole, ketoconazole and flucytosine). We compared this colorimetric method with a standard broth microdilution test, performed according to US National Committee for Clinical Laboratory Standards document M27-A guidelines, for determining the in-vitro susceptibilities of 180 isolates of Candida spp. (50 Candida albicans, 50 Candida glabrata, ten Candida kefyr, 20 Candida krusei, ten Candida lusitaniae, 20 Candida parapsilosis and 20 Candida tropicalis) and 20 isolates of Cryptococcus neoformans. The overall agreement between the results of the two methods (colorimetric MICs within +/- two log2 dilutions of standard MICs) were 99% for amphotericin B, 96.5% for flucytosine, 93% for itraconazole, 91.5% for fluconazole and 85.5% for ketoconazole. The overall levels of agreement between the two methods were > or = 94% for six of the eight species tested, the exceptions being C. neoformans and C. tropicalis where the overall agreement was 89% and 80% respectively. The poorest agreement between the results for individual agents was seen with C. tropicalis and the three azole agents (60-75% of colorimetric MICs within +/- two log2 dilutions of standard MICs), and C. neoformans for ketoconazole (50%). The Yeast One method appears to be a suitable alternative procedure for routine antifungal susceptibility testing of Candida spp. and C. neoformans.

Antifungal Agents↗

National surveillance of species distribution in blood isolates of Candida species in Japan and their susceptibility to six antifungal agents including voriconazole and micafungin.

OBJECTIVES: The aim of this study was to evaluate species distribution and antifungal susceptibility of Candida blood isolates in Japan. METHODS: In a 1 year surveillance programme, 535 Candida blood isolates were collected. Identification of species was followed by examination with the broth microdilution method, as described in NCCLS M27-A2, of antifungal susceptibility to six agents, including voriconazole and micafungin, with readings after 24 and 48 h of incubation. RESULTS: The overall species distribution was: 41% Candida albicans, 23% Candida parapsilosis, 18% Candida glabrata, 12% Candida tropicalis and 2% Candida krusei. The concentrations of fluconazole necessary to inhibit 90% of the isolates (MIC(90)) at 24/48 h were 0.25/1 mg/L for C. albicans, 0.5/2 mg/L for C. parapsilosis, 4/32 mg/L for C. glabrata and 4/>128 mg/L for C. tropicalis. Percentages of fluconazole resistance were 1.8% for C. albicans, 0.8% for C. parapsilosis, 5.2% for C. glabrata and 3.2% for C. tropicalis, taking the tendency of trailing growth of C. tropicalis into account. MIC(90) of voriconazole was 0.5 mg/L, although 35% of isolates less susceptible (>/=16 mg/L) to fluconazole showed resistance (>/=2 mg/L). Micafungin was very active against all species (MIC(90), 0.03 mg/L) except for C. parapsilosis (MIC(90), 2 mg/L). CONCLUSIONS: These data suggest that, in Japan, the species distribution of Candida bloodstream infections and the fluconazole resistance rate are similar to those reported previously in North America and Europe. Voriconazole and micafungin appear to have strong in vitro activity against Candida blood isolates, although continuing surveillance and further clinical research are needed.

Amphotericin B↗

[Involvement of secretory Candida proteinases in the adherence of C. tropicalis blastoconidia in a cell culture model].

The influence of the heterologous acid secretory Candida proteinases on the adherence of the non-proteinase secreting strain of C. tropicalis DSM 4959 to epitheloid cells (vero line) was examined. The proteinases of the following Candida strains were used: C. albicans ATCC 10261 (serotype A), C. albicans ATCC 48867 (serotype B), C. tropicalis DSM 4238. The assays were performed with the previously described in-vitro-adherence test [1] using the following principle steps: Candida proteinases and C. tropicalis blastoconidia were incubated with verocells in microtest plates in phosphate-buffer in the range of pH 4.0 to pH 7.0. Adherent Candida cells were detected according to Filler et al. [2] with anti-Candida-mannoprotein antibodies and a secondary anti-rabbit-peroxidase conjugate. Compared to controls with denaturated proteinases, the photometric evaluation of adherent C. tropicalis cells showed, under optimal conditions, an augmentation of the adherence due to the Candida proteinases of about 50%. The optimum of this adherence augmentation was in the range of pH 5.5 which is outside the general activity optimum of Candida proteinases (pH 3). The degree of purity of these proteinases had no marked influence on the adherence. The specificity of the proteinase dependent adherence augmentation could be demonstrated with the enzyme inhibitor Pepstatin A. C. tropicalis blastoconidia supplemented by pepstatin A and active Candida proteinase adhered in the same range as with denaturated proteinases in control tests. Our results suggest a function of Candida proteinases in the adherence process of blastoconidia to epithelia.

Animals↗

Candida species in stool, symptoms and complaints in general practice--a cross-sectional study of 308 outpatients.

Patients with unspecific symptoms were sometimes tested for Candida. In case of findings of this yeast-like fungus in their stools they often were labelled with the diagnosis of a "Candida-syndrome". This comprises headache, weakness, flatulence, ravenous appetite for sweets, itching skin and several more unspecific symptoms. All 500 randomly sampled patients in 12 mainly naturopathic practices were asked to take part. In case of participation, they received stool-tubes and questionnaires to be answered during the waiting-time by the patients themselves. We asked for details of lifestyle, diseases and a number of unspecific symptoms. The stool-tubes were sent to a microbiological lab within 24 h after being filled. About one-third of all 308 participants carried Candida albicans in their stools. This finding is regarded as normal. Smoking habits were highly associated to Candida: 45 of the 78 smokers (58%), but only 68 of the 230 (29%) non-smokers were Candida positive, P < 0.0001. Three more results were associated with Candida-positive stools: Candida-vaginitis, allergies against food and allergies in general. Hints of a Candida-syndrome could not be found. The relation with smoking cigarettes is a new result. Associations to Candida-vaginitis and allergies were described before.

Adolescent↗

DNA probes for the rapid identification of medically important Candida species using a multianalyte profiling system.

Recently, a new flow cytometric technology to detect multiple DNA target sequences in a single microtiter well plate was developed [multianalyte profiling (MAP) System, Luminex Corp., Austin, TX]. DNA probes, directed to the internal transcribed spacer 2 region of ribosomal DNA, were therefore designed to detect and differentiate PCR amplicons from six medically important Candida species using this system. Each probe was covalently linked to one of 100 available microsphere (bead) sets. Biotinylated PCR amplicons were then hybridized to the complementary probe on each bead set. Bound amplicons were detected fluorometrically using a streptavidin-linked reporter dye, R-phycoerythrin. Specific hybridization was noted for all six Candida species probes (mean sample-to-background ratio+/-standard error: Candida albicans, 58.7+/-1.2; Candida tropicalis, 53.2+/-3.8; Candida glabrata, 46.9+/-2.1; Candida parapsilosis, 59.9+/-1.6; Candida krusei, 54.7+/-3.7 vs. 0.9+/-0.03 for all heterologous Candida species DNA targets and vs. 1.0+/-0.1 for samples containing water instead of DNA; P < 0.001). The limit of test sensitivity was 0.5 pg of DNA. A sample could be processed and analyzed within 1 h post-PCR amplification. Therefore, the multianalyte profiling system was rapid, sensitive and specific for the detection and differentiation of the most medically important species of Candida.

Candida↗

Carriage of Candida species in the oral cavity in diabetic patients: relationship to glycaemic control.

To study the possible relationship between the quality of glycaemic control in diabetes mellitus and the carriage of Candida species, the candidal carrier status of 412 diabetic patients was examined using an oral rinse technique and correlated with measurements of random blood glucose and total glycosylated haemoglobin. Candida was isolated in 210 diabetics (51%) with 13 patients (6%) carrying more than one species. The positive isolates were: Candida albicans (89%), Candida krusei (2.8%), Candida glabrata (2.8%), Candida tropicalis (6.2%), Candida stellatoidea (2.8%) and Candida parapsilosis (0.5%). No association was identified between carriage rates and the type of treatment of diabetes, or with the quality of glycaemic control. As in non-diabetic subjects, the carriage rates were higher in diabetic patients wearing dentures. Thus, the oral carriage of Candida in diabetic patients was independent of glycaemic control but in certain sub-groups the carriage rates were higher, and involved uncommon candidal species.

Blood Glucose↗

Mice immunized by primary vaginal Candida albicans infection develop acquired vaginal mucosal immunity.

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

Animals↗

Differential cytokine production and Toll-like receptor signaling pathways by Candida albicans blastoconidia and hyphae.

Toll-like receptors (TLR) are crucial for an efficient antifungal defense. We investigated the differential recognition of blastoconidia and hyphae of Candida albicans by TLRs. In contrast to Candida blastoconidia, which stimulated large amounts of gamma interferon (IFN-gamma), the tissue-invasive Candida hyphae did not stimulate any IFN-gamma by human peripheral blood mononuclear cells (PBMC) or murine splenic lymphocytes. After stimulation with blastoconidia, the production of IFN-gamma was TLR4 dependent, as shown by the significantly decreased IFN-gamma production in anti-TLR4-treated PBMC and in splenic lymphocytes from TLR4-defective ScCr mice. In addition, peritoneal macrophages from ScCr mice produced less tumor necrosis factor alpha (TNF-alpha) than macrophages of control mice did when stimulated with Candida blastoconidia, but not with hyphae, indicating that TLR4-mediated signals are lost during hyphal germination. In contrast, macrophages from TLR2 knockout mice had a decreased production of TNF-alpha in response to both Candida blastoconidia and hyphae. Candida hyphae stimulated production of interleukin-10 through TLR2-dependent mechanisms. In conclusion, TLR4 mediates proinflammatory cytokine induction after Candida stimulation, whereas Candida recognition by TLR2 leads mainly to anti-inflammatory cytokine release. TLR4-mediated proinflammatory signals are lost during germination of Candida blastoconidia into hyphae. Phenotypic switching during germination may be an important escape mechanism of C. albicans, resulting in counteracting host defense.

Animals↗

Identification of Candida species by PCR and restriction fragment length polymorphism analysis of intergenic spacer regions of ribosomal DNA.

The PCR was used to amplify a targeted region of the ribosomal DNA from 84 Candida isolates. Unique product sizes were obtained for Candida guilliermondii, Candida (Torulopsis) glabrata, and Candida pseudotropicalis. Isolates of Candida albicans, Candida tropicalis, Candida stellatoidea, Candida parapsilosis, and Candida krusei could be identified following restriction digestion of the PCR products.

Base Sequence↗

Simple and rapid detection of Candida albicans DNA in serum by PCR for diagnosis of invasive candidiasis.

A rapid and sensitive PCR assay for the detection of Candida albicans DNA in serum was established. DNA from human serum samples was purified using the QIAamp blood kit, which proved to be a fast and simple method for isolating minute amounts of Candida DNA from clinical specimens for diagnosis of invasive candidiasis. Universal primer sequences used in the PCR assay are derived from the internal transcribed spacer rRNA gene of fungi, whereas the biotinylated hybridization probe used in a DNA enzyme immunoassay (DEIA) binds specifically to C. albicans DNA. The sensitivity of this PCR-DEIA method is very high; the detection limit for genomic Candida DNA is one C. albicans genome per assay. Blood from uninfected and infected persons, ranging from healthy volunteers, patients with mucocutaneous infections, and patients at risk to develop a systemic Candida infection to patients with an established systemic candidiasis, was analyzed for the presence of C. albicans to diagnose fungal infection. Candida DNA could not be detected in sera of 16 culture-negative controls and from 11 nonsystemic candidal infections by PCR or DEIA. Blood cultures from patients at risk were all negative for Candida, whereas all blood cultures from systemic candidiasis patients were positive. However, Candida DNA could be detected by PCR and DEIA in the serum from three out of nine patients who were at risk for a systemic infection and in the serum of all seven patients who had already developed an invasive Candida infection. PCR is more sensitive than blood culture, since some of the patients at risk for invasive yeast infection, whose blood cultures were all negative for Candida, tested positive in the PCR amplification. These results indicate the potential value of PCR for detecting C. albicans in serum samples and for identifying patients at risk for invasive candidiasis.

Candida albicans↗

Fluconazole and voriconazole multidisk testing of Candida species for disk test calibration and MIC estimation.

Fluconazole and voriconazole MICs were determined for 114 clinical Candida isolates, including isolates of Candida albicans, Candida glabrata, Candida krusei, Candida lusitaniae, Candida parapsilosis, and Candida tropicalis. All strains were susceptible to voriconazole, and most strains were also susceptible to fluconazole, with the exception of C. glabrata and C. krusei, the latter being fully fluconazole resistant. Single-strain regression analysis (SRA) was applied to 54 strains, including American Type Culture Collection reference strains. The regression lines obtained were markedly different for the different Candida species. Using an MIC limit of susceptibility to fluconazole of < or =8 microg/ml, according to NCCLS standards, the zone breakpoint for susceptibility for the 25-microg fluconazole disk was calculated to be > or =18 mm for C. albicans and > or =22 mm for C. glabrata and C. krusei. SRA results for voriconazole were used to estimate an optimal disk content according to rational criteria. A 5-microg disk content of voriconazole gave measurable zones for a tentative resistance limit of 4 microg/ml, whereas a 2.5-microg disk gave zones at the same MIC level for only three of the species. A novel SRA modification, multidisk testing, was also applied to the two major species, C. albicans and C. glabrata, and the MIC estimates were compared with the true MICs for the isolates. There was a significant correlation between the two measurements. Our results show that disk diffusion methods might be useful for azole testing of Candida isolates. The method can be calibrated using SRA. Multidisk testing gives direct estimations of the MICs for the isolates.

Antifungal Agents↗

Binding sites for LH in Candida albicans: comparison with the mammalian corpus luteum LH receptor.

We have described recently the presence of binding sites for human LH (hLH) and human chorionic gonadotrophin (hCG) in microsomal and cytosol fractions prepared from the dimorphic, pathogenic fungus, Candida albicans. We have now compared the properties of Candida LH/hCG-binding sites with those of the ovine luteal LH receptor. Sheep luteal LH-binding sites were associated with luteal membranes, and little or no binding activity was present in cytosol fractions. In contrast, significant LH/hCG-binding activity was present in Candida cytosol. Moreover, there were marked differences in sensitivity to inhibition by metal ions, association and dissociation rates, and affinity constants between sheep and Candida LH-binding sites. Scatchard plots of 125I-labelled hLH binding to sheep luteal receptors demonstrated a single high-affinity component (association constant (Ka) 0.3 litres/pmol) which was displaceable by hCG. In contrast, Scatchard plots of binding to Candida microsomes and cytosol fractions demonstrated two components, one with high affinity (Ka 0.18 litres/pmol) and low capacity and a second site with lower affinity (Ka 4 litres/nmol) and high capacity, both of which were displaceable by unlabelled hCG. Gel permeation chromatography of cytosol demonstrated two distinct peaks of LH-binding activity with approximate molecular weights of greater than 1,000,000 and 30,000-50,000. Scatchard plots of 125I-labelled hLH binding to the higher molecular weight peak demonstrated a single, high-affinity LH-binding site (Ka 0.18 litres/pmol), whereas the lower molecular weight fraction contained both high- (Ka 0.17 litres/pmol) and low-affinity (Ka 4 litres/nmol) LH-binding sites. Both partially purified and highly purified hCG and hLH preparations displaced binding of 125I-labelled hLH and hCG to sheep luteal LH receptors at similar concentrations. 125I-labelled hLH/hCG binding to Candida membranes was also displaceable by low levels (ng) of partially purified hCG preparations, but much higher levels (micrograms) of highly purified hLH and hCG were required. This paradoxical observation suggested the presence of radiolabelled contaminants, or damaged forms induced during radioiodination of hormone tracers, which can bind more strongly to Candida membranes than unlabelled hCG and hLH but which do not bind to sheep LH receptors. However, no evidence for hLH tracer contaminants with differential binding to Candida and sheep luteal receptors was obtained following gel exclusion chromatography or fractionation on Concanavalin A-Sepharose. (Although three distinct 125I-labelled hLH fractions were resolved on Concanavalin A-Sepharose, presumably reflecting differences in their carbohydrate compositions, all three tracer peaks bound equivalently to both Candida membranes and ovine luteal LH receptors).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Intestinal colonization with Candida albicans and mucosal immunity.

AIM: To observe the relationship between intestinal lumen colonization with Candida albicans and mucosal secretory IgA (sIgA). METHODS: A total of 82 specific-pathogen-free mice were divided randomly into control and colonization groups. After Candida albicans were inoculated into specific-pathogen-free mice, the number of Candida albicans adhering to cecum and mucosal membrane was counted. The lymphocyte proliferation in Peyer's patch and in lamina propria was shown by BrdU incorporation, while mucosal sIgA (surface membrane) isotype switch in Peyer's patch was investigated. IgA plasma cells in lamina propria were observed by immunohistochemical staining. Specific IgA antibodies to Candida albicans were measured with ELISA. RESULTS: From d 3 to d 14 after Candida albicans gavaging to mice, the number of Candida albicans colonizing in lumen and adhering to mucosal membrane was sharply reduced. Candida albicans translocation to mesenteric lymph nodes occurred at early time points following gavage administration and disappeared at later time points. Meanwhile, the content of specific IgA was increased obviously. Proliferation and differentiation of lymphocytes in lamina propria were also increased. CONCLUSION: Lymphocytes in lamina propria play an important role in intestinal mucosal immunity of specific-pathogen-free mice when they are first inoculated with Candida albicans. The decreasing number of Candida albicans in intestine is related to the increased level of specific IgA antibodies in the intestinal mucus.

Animals↗

[Candida in biological human samples].

Infections by Candida have been raising in the last decades, and risk factors, mainly immunosuppression and the appearance of Candida no albicans, are determinants in the prognosis of these mycoses. The purpose of this investigation was to identify and establish the prevalence of C. albicans and Candida spp. in candidiases, in patients to the Hospital Universitario de Maracaibo, whose biological samples were processed for both direct examination and cultures, needed for the proper identification. From October 1996 to October 1998, isolation and identification of yeasts of Candida were performed in 177 biological samples: 73 (41.24%) Candida albicans and 104 (58.75%) Candida spp. Both blastoconidias and pseudohyphae were found in 34 samples (19.21%), 24 of which (70.5%) were diagnosed as C. albicans and 10 (29.5%), as Candida spp. Blastoconidias identified by direct method were distributed as C. albicans 34.2% and Candida spp. 65.7%. C. albicans was found more often in intertrigo, sputum and in bronquial lavage samples. Candida spp. was more frequent in nails. Candidiasis is a frequently diagnosed mycosis in hospitals, mainly among immunossuppresed patients. It is very important to use direct microscopical evaluation and cultures, in order to establish the presence of blastoconidias and pseudohyphae, that will help to diagnose the aethiology and prevalence of candidiasis. It is also important to recognize subungueal candidiasis in hospital staff, that could spread the infection to inpatients.

Adult↗