Differences in fecal microflora between patients with atopic dermatitis and healthy control subjects

BACKGROUND: The prevalence of allergic diseases, such as atopic dermatitis (AD), has been increasing. However, few investigations have been made of the intestinal microflora in Japanese patients with AD. OBJECTIVE: The purpose of this study was to determine the differences in microflora, fecal serum IgA concentrations, and skin IgA contents between patients with AD and healthy control subjects. METHODS: This trial was conducted as a case-control study using 30 minor patients with AD and age- and sex-matched healthy control subjects (n = 68). One week after a questionnaire was administered, fecal specimens and 24-hour skin secretion specimens were collected from all subjects. Fecal microflora, fecal IgA concentrations, and IgA contents on the skin surface were analyzed. RESULTS: The counts of Bifidobacterium (in log10 colony-forming units per gram) were significantly lower in patients with AD than in healthy control subjects (9.75 +/- 0.68 vs 10.10 +/- 0.50 log(10) colony-forming units/g, P <.05). In particular, percentages of Bifidobacterium were significantly lower in patients with severe skin symptoms than in those with mild skin symptoms (40% +/- 6% vs 19% +/- 6%, P <.05). In addition, the frequency of occurrence of Staphylococcus was significantly higher in patients with AD than in healthy control subjects (83% vs 59%, P <.05). There were no significant differences in fecal IgA content or IgA content on the skin between the 2 groups. CONCLUSION: Patients with AD had lower counts of Bifidobacterium than healthy control subjects, and the frequency of Staphylococcus was higher in patients with AD than in control subjects. Disorder of the intestinal microflora might play a role in the onset of AD and the aggravation of skin symptoms.     

Investigation of Candida dubliniensis in Candida spp.-positive hemocultures

Candida dubliniensis is one of the Candida species which was first recognized in 1995. The yeast was misidentified because of its phenotypic similarities with Candida albicans. In this study, blood samples of patients from various departments at Ankara University Medical Faculty between January 1996 and September 2000 were investigated for distribution of Candida spp. and presence of C. dubliniensis. Ninety-eight culture positive fungi were included in the study. Phenotypic tests for identification of C. dubliniensis and tests for differentiation of the yeast from C. albicans, such as colony morphology on Staib agar, growth at 42 degrees C and 45 degrees C, beta-glucosidase activity and carbohydrate assimilation, were carried out. Sixty-four of the isolates produced germ tubes and chlamydospores, and none of them had the phenotypic characteristics of C. dubliniensis. Further large-scale studies of specific patient groups are necessary to reveal the etiologic importance of this yeast.     

A study of human fecal microbiocenosis by experimental in vitro modeling of constipation

A study of human fecal microbiocenosis using experimental in vitro modeling of constipation was carried out. The study revealed certain dynamics of the incidence, proportion and ratio of microorganisms isolated from the feces at various stages of cultivation. The character of the changes of these parameters depended on the type of the microorganism. According to the results of the experiment, transitory flora (staphylococci; candidas) begins to disappear within the first day of cultivation. If intestinal evacuation is retarded for more than 2 or 3 days, distortion of the microbiological characteristics of the residential microflora, bifidobacteria and escherichiae, is possible. These processes may lead to a false positive result of dysbiosis analysis. The authors conclude that dysbiotic changes in constipation are caused by fecal retention. That is why correct diagnostics of dysbiosis is possible only after regular everyday stool is reestablished.     

Rapid identification of Candida spp. in peritonitis patients by Raman spectroscopy

This prospective study evaluated Raman spectroscopy for the identification of clinically relevant Candida spp. in peritonitis patients. A Raman database was developed by measuring spectra from 93 reference strains belonging to ten different Candida spp. Clinical samples were obtained from the surgical department and intensive care unit of a tertiary university hospital. In total, 88 peritoneal specimens from 45 patients with primary, secondary or tertiary peritonitis were included. Specimens were cultured initially on a selective Sabouraud medium that contained gentamicin to suppress bacterial growth. For conventional identification, a chromogenic medium was used for presumptive identification, followed by use of the Vitek 2 system for definitive identification (requiring a total time of 48-96 h). Raman measurements were taken on overnight cultures from Sabouraud-gentamicin medium. Thirty-one samples were positive for Candida by culture. Using multivariate statistical analyses, a prediction accuracy of 90% was obtained for Raman spectroscopy, which appears to offer an accurate and rapid (12-24 h) alternative for the identification of Candida spp. in peritonitis patients. The reduced turn-around time is of great clinical importance for the treatment of critically ill patients with invasive candidiasis in intensive care units.     

False-positive antiglobulin tests in healthy subjects and in hospital patients.

An antiglobulin reagent containing anti-IgG, -C4c, -C4d, -C3c, and -C3d was used to test red blood cells from healthy subjects and from patients in hospital. In tests read macroscopically on opal tiles, no positive reactions were found in the healthy subjects. Positive reactions were, however, observed in 7% of the hospital patients. Further tests with monospecific reagents showed the positive reactions to be due to the presence of C4d and C3d on the red cells. All patients with positive antiglobulin reactions had serious disease, in most cases associated with abnormal antibody production or abnormal immunoglobulin levels.     

Elevated fecal Candida counts in patients with antibiotic-associated diarrhea: role of soluble fecal substances

To assess the role of soluble fecal substances in the elevation of fecal Candida counts in patients with antibiotic-associated diarrhea (AAD), we investigated the growth of Candida albicans in vitro in serially diluted stool fluids from patients with AAD and healthy subjects. There were significantly higher Candida albicans counts in stool fluids diluted 1:10 from AAD patients than in healthy subjects and the phosphate-buffered saline growth control, which may be due to reduced soluble Candida inhibitors and increased availability of growth factors and nutrients.     

Involvement of Fusarium spp. in fungal keratitis

Members of the filamentous fungal genus Fusarium are among the agents most frequently causing keratomycosis in humans. Fusarium keratitis is most common among agricultural workers in geographical regions with hot, humid, tropical or semi-tropical climates, but can occur more rarely in countries with temperate climates, such as Hungary. Keratitis is usually treated with a topical antifungal agent, sometimes in combination with sub-conjunctival injections and/or antimycotic agents, but therapeutic keratoplasty may be needed for patients whose corneal infection does not resolve. Early and accurate diagnosis, coupled with appropriate antifungal therapy, is crucial for improving the chances of complete recovery.     

Potential risk factors for infection with Candida spp. in critically ill patients

The incidence, risk factors and prognostic factors for candidal infection were determined in a prospective study of 280 infected patients. Thirty-one (11%) patients were infected with Candida spp., sub-divided into 18 (58%) with C. albicans, and 13 (42%) with non-albicans spp. (six C. glabrata, three C. parapsilosis, and one each of C. krusei, C. tropicalis, C. guilliermondii and C. lusitaniae). Infection with Candida spp. was always associated with concurrent bacterial infection. By univariate logistic regression analysis, the degree of morbidity and the duration of mechanical ventilation were independent predictive factors for death, but infection with Candida spp., was not. Factors associated with Candida spp. infection were the degree of morbidity, intensive care unit length of stay, alterations of immune response, and the number of medical devices involved. By multivariate logistic regression analysis, the only independent risk factor for candidal infection was intensive care unit length of stay.     

Candida orthopsilosis and Candida metapsilosis spp. nov. to replace Candida parapsilosis groups II and III

Two new species, Candida orthopsilosis and C. metapsilosis, are proposed to replace the existing designations of C. parapsilosis groups II and III, respectively. The species C. parapsilosis is retained for group I isolates. Attempts to construct a multilocus sequence typing scheme to differentiate individual strains of C. parapsilosis instead revealed fixed DNA sequence differences between pairs of subgroups in four genes: COX3, L1A1, SADH, and SYA1. PCR amplicons for sequencing were obtained for these four plus a further seven genes from 21 group I isolates. For nine group II isolates, PCR products were obtained from only 5 of the 11 genes, and for two group III isolates PCR products were obtained from a different set of 5 genes. Three of the PCR products from group II and III isolates differed in size from the group I products. Cluster analysis of sequence polymorphisms from COX3, SADH, and SYA1, which were common to the three groups, consistently separated the isolates into three distinct sets. All of these differences, together with DNA sequence similarities <90% in the ITS1 sequence, suggest the subgroups should be afforded species status. The near absence of DNA sequence variability among isolates of C. parapsilosis and relatively high levels of sequence variability among isolates of C. orthopsilosis suggest that the former species may have evolved very recently from the latter.     

Confirmation of associations between ion channel gene SNPs and QTc interval duration in healthy subjects

Population-based association studies have identified several polymorphic variants in genes encoding ion channel subunits associated with the electrocardiographic heart-rate-corrected QT (QTc) length in healthy populations of Caucasian origin (KCNH2 rs1,805,123 (K897 T) and rs3,815,459, SCN5A rs1,805,126 (D1,819D), 1,141-3 C>A, rs1,805,124 (H558R), and IVS24+116 G>A, KCNQ1 rs757,092, KCNE1 IVS2-128 G>A and rs1,805,127 (G38S), and KCNE2 rs2,234,916 (T8A)). However, few of these results have been replicated in independent populations. We tested the association of SNPs KCNQ1 rs757,092, KCNH2 rs3,815,459, SCN5A IVS24+116 G>A, KCNE1 IVS2-128 G>A and KCNE2 rs2,234,916 with QTc length in two groups of 200 subjects presenting the shortest and the longest QTc from a cohort of 2,008 healthy subjects. All polymorphisms were in Hardy-Weinberg equilibrium in both groups. The minor allele SCN5A IVS24+116 A was more frequent in the group of subjects with the shortest QTc, whereas the minor alleles KCNQ1 rs757,092 G and KCNH2 rs3,815,459 A were more frequent in the group with the longest QTc. There was no significant difference for KCNE1 IVS2-128 G>A and KCNE2 rs2,234,916 between the two groups. Haplotype analysis showed a twofold increased risk of QTc lengthening for carriers of the haplotype, combining alleles C and A of the two common KCNE1 SNPs, IVS2-129 C>T (rs2,236,609) and rs1,805,127 (G38S), respectively. In conclusion, our study confirms the reported associations between QTc length and KCNQ1 rs757,092 and KCNH2 rs3,815,459.     

Genetic dissimilarity of commensal strains of Candida spp. carried in different anatomical locations of the same healthy women.

Candida spp. carriage and strain relatedness were assessed in 52 healthy women at 17 anatomical locations by using an isolation procedure which assesses carriage intensity and by using a computer-assisted DNA fingerprinting system which computes genetic similarity between strains on the basis of the patterns of Southern blots probed with the moderately repetitive sequence Ca3. Candida spp. were cultured from 73% of the test individuals, most frequently from the oral (56%), vulvovaginal (40%), and anorectal (24%) regions. Half of the test individuals with Candida spp. carried the organism simultaneously in more than one of the three general areas of carriage. Isolates from different body locations of the same individual were either completely unrelated, identical, or highly similar but nonidentical. In 11 cases in which Candida spp. were simultaneously isolated from the oral cavity and vaginal canal, seven pairs of isolates were genetically unrelated and four pairs were similar but nonidentical. In the latter cases, the isolate pairs each appear to have arisen by genetic divergence from a single progenitor. A comparison of the genetic relatedness of isolates from different individuals further uncovered a single strain which was vaginospecific in the Iowa City, Iowa area and reduced genetic diversity among vulvovaginal strains compared with those isolated from other body locations. These results suggest that strains adapt to different anatomical locations and, conversely, that in a healthy individual there is anatomical selection of vaginotropic, anotropic and orotropic strains of Candida spp.     

Anti-IL-1 alpha autoantibodies in patients with rheumatic diseases and in healthy subjects.

Antibody reactivity against the 'mitochondrial M2 antigen' was determined in sera from 10 patients with primary biliary cirrhosis (PBC), using Western blotting after SDS-PAGE separation of rat liver mitochondria (RLM) and plasma membrane proteins. The molecular weights of the major M2 antigens in rat liver mitochondria were 67 and 50 kD. Two of the 10 PBC patients did not react to any of these major antigens, eight reacted to the 67-kD and four of those also to the 50-kD antigen. The 67- and 50-kD antigens were present in both plasma membrane and RLM and had affinity to concanavalin A. Antibody reactivity against the 67-kD antigen could be detected in both IgG and IgA as well as in the IgM class. The reactive IgG subclasses to both types of antigen preparations were mainly of the G1 and G3 isotypes. This reactivity was always stronger with antigens from the plasma membrane preparations. Sera from two patients with high antibody titres against mitochondria also reacted with IgG2 against the 50-kD antigen from plasma membrane, but not to the corresponding antigen in mitochondria. Reactivity of antibodies in PBC sera to the periphery of viable hepatocytes and radioactive surface labelling of the 50-kD component are both consistent with a plasma membrane localization of M2. Serum from healthy controls and several patients with different diseases did not contain antibodies reactive against any of the antigens described. We suggest that antigens, partly identical to the mitochondrial M2, are located in the plasma membrane compartment. The PBC pathogenetical consequences of these findings are discussed.     

Exposure to light in healthy elderly subjects and Alzheimer's patients

Exposure to light was recorded from 10 healthy elderly adults and 13 age-matched subjects with senile dementia of the Alzheimer's type (SDAT). Data were recorded in the home, for an average of 5 days, while subjects continued their normal daily activities. Subjects were exposed to remarkably small intervals of illumination exceeding 2000 lux. Subjects with SDAT were exposed to bright light significantly less than healthy controls (0.5 vs. 1.0 hr). Whether or not they had SDAT, males were exposed to illumination exceeding 2000 lux significantly more than were females. Healthy elderly received about two-thirds the duration of bright light received by healthy younger subjects. These findings suggest an association between decreased exposure to bright light and the declines in sleep quality which typically accompany normal and pathological aging.     

Phenotypic characterization and DNA relatedness in human fecal isolates of Aeromonas spp.

Phenotypic characteristics were used to identify 189 Aeromonas strains isolated from human feces. One hundred forty-two of these strains were placed in 11 DNA hybridization groups, and the genetic and phenotypic data were compared. According to the criteria of Popoff, 66% of the strains were identified as Aeromonas caviae, 18% were identified as A. sobria, and 16% were identified as A. hydrophila. Some biochemical characteristics differed from the criteria of Popoff; 19 of 40 (48%) of tested strains were encapsulated, 42 of 124 (34%) of A. caviae strains were nonmotile, and all A. sobria strains were resistant to KCN. Gas production from D-glucose was temperature dependent; 11 of 64 (17%) A. hydrophila and A. sobria strains produced gas only at 22 degrees C. Of 142 Aeromonas strains, 57% belonged to hybridization group 4, 25% belonged to group 8, 11% belonged to group 1, 4% belonged to group 5A, 2% belonged to group 3, and 1% belonged to group 2. Of 26 strains phenotypically identified as A. hydrophila, 8 (31%) were in hybridization group 8, which contains strains of the new species A. veronii. It therefore appears that our ability to identify Aeromonas strains phenotypically is not sufficiently specific. Either additional definitive biochemical markers must be found or phenotypic identification, at least for some Aeromonas groups, must be regarded as only presumptive.     

Detection of caspofungin resistance in Candida spp. by Etest

The caspofungin susceptibilities of 28 Candida sp. clinical isolates, including 8 caspofungin-resistant isolates characterized by mutations in the Fks1 protein, were determined by the Etest in RPMI and AM3 media. Good discrimination between wild-type and mutant isolates was obtained. These results suggest that the Etest is valuable for the detection of caspofungin resistance in Candida spp.     

Serum ferritin in healthy subjects and type 2 diabetic patients

In order to study the relationship between the serum ferritin level and the components of the insulin resistance syndrome in type 2 diabetic patients, we evaluated fifty type 2 diabetic patients who were selected according to NDDG/WHO criteria from those patients attending Korea University Hospital from 1997 to 1998. Twenty-five healthy non-diabetic subjects of comparable age and sex distribution acted as a control group. The results showed that the value of log ferritin was higher in the type 2 diabetes patients than the control subjects, but not at a statistically significant level (p = 0.09). Log ferritin was correlated with fasting blood sugar level (r = 0.235, p = 0.048) and body mass index (BMI) (r = 0.285, p = 0.05). In the type 2 diabetic patients, log ferritin was correlated with fasting C-peptide (r = 0.478, p = 0.009). In the control subjects, log ferritin was correlated only with BMI (r = 0.477, p = 0.012). In a stepwise multiple regression analysis, the diabetic group showed a significant correlation between fasting C-peptide and log ferritin (p = 0.001). In the control group, the fasting sugar level was significantly correlated with log ferritin (p = 0.034). These results suggest that serum ferritin can be employed as a marker of not only glucose homeostasis but also insulin resistance both in type 2 diabetic and control subjects.     

Histamine binding proteins identified in healthy subjects and asthmatic patients

Histamine-binding proteins have been isolated from healthy subjects and patients with endogenous bronchial asthma. Histaminopectic properties were exhibited mainly by IgA, IgG, IgM immunoglobulins, orosomucoid, albumin and ceruloplasmin. Quantitative determination of the identified proteins showed that in the control group immunoglobulins bound histamine in 97.5%, whereas in asthmatics the corresponding percentage (about 81%) was statistically significantly lower.