SUMMARYAim: To compare the results of culture and epsilometer test with fluorescence in situ hybridization for the detection of Helicobacter pylori and the presence of clarithromycin-susceptible and clarithromycin-resistant strains in antral biopsies from children. Methods: Antral biopsies from 149 unselected children were investigated prospectively; 15 had previously received anti-H. pylori therapy. H. pylori status was defined by histology, rapid urease test and 13 C-urea breath test. Fluorescence in situ hybridization was applied on fresh tissue with probes specific for the clarithromycin-susceptible wild type and three clarithromycin-resistant mutants. Susceptibility to clarithromycin was tested by epsilometer test in two laboratories.Results: Culture and fluorescence in situ hybridization gave negative results in all 66 H. pylori-negative children (specificity, 100%). Of 83 infected children, cultures were successful in 75 (90%), epsilometer test in 71 (86%) and fluorescence in situ hybridization in 77 (93%). Eleven children (13%) showed discrepant results between the applied methods, indicating mixed infection. Clarithromycin-resistant isolates were identified in 16 of 73 previously untreated children. Conclusions: Primary resistance to clarithromycin is common (22%) in H. pylori isolates from children living in Germany. Fluorescence in situ hybridization is an excellent, fast method for the detection of H. pylori and clarithromycin-resistant mutants in gastric biopsies. Multiple biopsies identify mixed infections, indicating that clarithromycin-resistant and clarithromycin-susceptible strains are not evenly distributed within the stomach.

We report on the successful application of fluorescent in situ hybridization for detection of Helicobacter pylori and determination of its clarithromycin susceptibility in formalin-fixed and paraffin-embedded gastric biopsy specimens that had been prepared for pathological examination. This method is useful when results from conventional culturing with antibiotic susceptibility testing are not available.Clarithromycin resistance in Helicobacter pylori is based on single-base mutations within the peptidyltransferase-encoding region of the 23S rRNA gene (4,7,9). At the Max von Pettenkofer-Institute, we have established the fluorescent in situ hybridization (FISH) technique with rRNA-targeted fluorescence-labeled oligonucleotide probes specific for three described mutations in which the adenine residues at positions 2143 and 2144 are replaced by guanine (A2143G and A2144G) and cytosine (A2143C), respectively (8). Recently, we have demonstrated the application of FISH for (i) simultaneous detection of H. pylori and identification of the 23S rRNA point mutations responsible for macrolide resistance directly from shock-frozen gastric biopsy specimens (5) and (ii) rapid and accurate determination of genotypic clarithromycin resistance in cultured H. pylori isolates (6). FISH has been implemented at the Max von Pettenkofer-Institute as a work technique that is part of the daily routine and was found to be a more sensitive and reliable method than conventional culturing for detection of H. pylori in gastric biopsy specimens (5).Ideally, antral biopsy specimens taken during diagnostic endoscopy from patients with upper abdominal complaints are sent to (i) a microbiology laboratory for culturing of H. pylori and subsequent susceptibility testing with all relevant antibiotic drugs and to (ii) a pathologist for histological diagnosis of gastritis according to the updated Sydney system and histological determination of H. pylori infection by adequate staining techniques such as those that use the Wharthin-Starry or modified Giemsa stain. The "gold standard" for accurate diagnosis of an H. pylori infection is either culturing of the pathogen and/or concordant positive results obtained by histology and the rapid urease test or the 13 C-urea breath test (UBT), since none of these diagnostic tests except bacterial culturing are 100% specific. The major advantage of FISH is the fact that the rRNA-targeted fluorescence-labeled oligonucleotide probes can be used for accurate determination of macrolide susceptibility, thus providing the clinician with important information with which to make a proper treatment recommendation. Clarithromycin is a key component of most triple therapies for eradication of H. pylori (2), and the resistance of the pathogen to this antibiotic is an important cause of treatment failure, particularly in children, who are reported to harbor H. pylori strains with a high rate of primary resistance to this macrolide (1, 3). The aim of this retrospective study was to evaluate the sensitivity and specificity of F...

Research on identifying the relevant risk factors for suicides is faced with a multitude of methodological problems. The present study attempts to improve on some of these problems and to isolate those risk factors that are accessible in the early stages of the treatment of inpatients. A total of 3792 inpatients with monopolar or bipolar depression were treated during the period 1981-1992. Suicides (n=33) and controls (n=3759) were compared with respect to 77 sociodemographic and anamnestic variables and 195 standardised items of the admission summary. In addition to an analysis of contingency tables a discriminant analysis was performed. The suicide rate of patients with depressive psychosis was 2.7 times higher than the average rate of 0.324% for the entire clinic. Suicidal tendencies on admission proved to be the best predictor with a frequency of 91% in the suicide group and 40% in the control group, previous attempted suicide being the second best predictor. We conclude that the rate of inpatient suicide may have been underestimated for methodological reasons in the past decades. Many of the risk factors discussed in the literature may be of little predictive value at least in the initial stages of hospital treatment.

Monocyte adhesion to VSMCs induces TF mRNA and protein expression and monocyte PCA, which is regulated by beta2-integrin-mediated monocyte adhesion to ICAM-1 on VSMCs.