Hemolytic properties and riboflavin synthesis of Helicobacter pylori: cloning and functional characterization of the ribA gene encoding GTP-cyclohydrolase II that confers hemolytic activity to Escherichia coli

Various strains of Helicobacter pylori were able to lyse erythrocytes from sheep, horse, and human when grown on blood agar. The hemolysis did not depend on the production of the vacuolating cytotoxin VacA as demonstrated by the hemolytic behavior of an isogenic vacA-negative mutant strain. The hemolytic activity could be detected in cell-free supernatants and was not regulated by iron. To isolate genes coding for proteins involved in the destruction of erythrocytes, a plasmid-based DNA library was screened for expression of lytic activity on blood agar. This approach revealed that the H. pylori ribA gene confers hemolytic properties to Escherichia coli. The ribA gene encodes the enzyme GTP-cyclohydrolase II [EC 3.5.4.25] that catalyzes the initial step in the synthesis of riboflavin. The predicted amino acid sequence of the H. pylori RibA protein showed a high degree of similarity to equivalent enzymes from microorganisms and from plants. The single gene on a plasmid restored riboflavin synthesis in a ribA mutant of E. coli and induced hemolytic activity. Furthermore, ribA overexpression was associated with the production of a fluorescent yellow molecule that was not identical with riboflavin. Hemolysis was also seen for the ribA gene from E. coli, indicating that this feature was not specific for the H. pylori gene. The presence of ribA in various H. pylori strains was confirmed by Southern blot hybridization and by polymerase chain reaction with specific primers. This analysis revealed that microdiversity exists within the DNA region upstream from ribA, which was further confirmed by nucleotide sequence analysis. Received: 25 July 1997     

The effect of physeal traction applied to the triradiate cartilage on acetabular growth

Summary Physeal distraction was applied with an external fixator to the triradiate cartilage of dogs with the aim of increasing the capacity of the acetabulum. The force was continued for from 2 to 6 weeks and the consequent changes were evaluated with regard to function and structure by radiography and microscopy. The distraction, without producing epiphysiolysis and destruction of the cartilage, resulted in expansion of the pelvic bones. The depth and volume of the acetabulum were increased, but the acetabular angle was decreased. Distraction also caused proliferation of the lacunar cells and the number of mammillary processes in the cartilage columns increased. Distraction can therefore be applied to the triradiate cartilage to enlarge the capacity of the acetabulum without producing epiphysiolysis.

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Résumé Dans cette étude une distraction a été appliquée sur le cartilage en Y. Douze chiens bâtards, âgés de 2.5 à 4 mois, ont été utilisés pour cette expérimentation. Une force de distraction de 80 Newton a été appliquée d'emblée sur le cartilage en Y. L'application de cette force a été poursuivie sur les animaux pendant 2, 4 ou 6 semaines. A la fin de cette période, les changements de la cavité cotyloïde et du cartilage en Y ont été évalués sur le plan fonctionnel, morphologique, radiologique et histopathologique Aucune altération fonctionnelle n'a été mise en évidence dans les groupes opérés par rapport au groupe de contrôle. On a observé que la distraction a déterminé une expansion massive de l'ilion, du pubis et de l'ischion. Sur les hanches de contrôle la profondeur de la cavité cotyloïde a été évaluée à 13.5 mm, le volume à 1.96 cc et l'angle de la cavité cotyloïde à 29.9°. Après distraction ces valeurs ont été respectivement de 14.4 mm, de 2.10 cc et de 25.7°. La distraction entraîne donc un accroissement de profondeur et de volume de la cavité cotyloïde, mais inversement une diminution de l'angle de cette cavité. Ces résultats montrent que la distraction détermine une prolifération des cellules lacunaires et un accroissement des processus mammaires dans les colonnes du cartilage en Y. Ils montrent également qu'elle peut augmenter la taille de l'acetabulum.

     

Renal allograft immunosuppression

We have investigated the impact of triple drug immunosuppression on the occurrence of early inflammatory episodes, as detected by fine needle aspiration biopsy, and of episodes of clinical rejection during the immediate postoperative period. The prospective component of this study includes 128 consecutive first cadaveric renal transplant recipients receiving triple drug treatment consisting of azathioprine (Aza), cyclosporin (CyA) and methylprednisolone (MP). For controls we have used three historical groups: one immunosuppressed with Aza and MP (group A), another with CyA monotherapy (group B), and the third with CyA together with MP (group C) in equivalent drug dosages. On the average, 0.8 episodes of inflammation per patient were recorded during the immediate postoperative period of 30 days with triple drug treatment. This was significantly less than the 1.3 episodes in patients receiving Aza and MP (P<0.01), the 1.7 episodes in patients on CyA monotherapy (P<0.001), or the 1.6 episodes in patients receiving CyA together with MP (P<0.001). Although the first episode of inflammation commenced concurrently in each group and the peak intensity of inflammation was the same, the mean duration of inflammation was significantly shorter-2.7 days-under triple drug treatment than the 7.8–11.7 days for controls (P<0.001). The frequency of rejection episodes under triple treatment was also significantly lower-0.2 per patient-than the 0.8 per patient in controls (P<0.001). The first rejection episode occurred later in the triple drug treatment group-on the average, on day 15.2-than in the historical controls (on days 7.7–11.7). There was, however, no difference in the duration of rejection. There were no differences in patient survival between the four groups. Graft survival was 97% at 10 weeks for triple drug-treated recipients and 79%, 68%, and 87% for first grafts in groups A, B, and C, respectively. Disregarding a minor demographic bias for the triple drugtreated group with respect to preformed antibodies and preoperative dialysis treatment, the study suggests that the triple drug protocol, in the short run, is superior to any conceivable double drug combination or CyA monotherapy.     

Destructive changes of the spine in magnetic resonance imaging

Summary Twenty-nine patients were examined by magnetic resonance imaging for various lesions of the spine. The results of these scans were compared with those of plain radiographs, computertomographs, and radionuclide bone scans. The findings were substantiated by intraoperative or histological findings in 18 cases. The MRI scans proved to be very sensitive in the detection of a wide spectrum of morphological changes of the bone marrow of vertebral bodies. Characteristic changes of the signal patterns for inflammatory and tumorous lesions were not observed. The differentiation of these lesions will presently continue to have to be based on morphological criteria.No benefits in any form have been received or will be received from a commercial party related directly or indirectly to the subject of this study. No funds were received in support of this study     

Mechanical energy absorption in human hand-arm exposed to sinusoidal vibration

Summary A possible basis for risk assessment of human exposure to vibration when using hand-held tools may be to determine the amount of mechanical energy that is absorbed by the hand-arm system. The aim of this investigation was to study the absorption of mechanical energy in the human hand-arm system during exposure to sinusoidal vibration within the frequency range of 20 to 1500 Hz. A handle, specially designed for this type of experiments, was used during the measurements. The influence of various experimental conditions, such as three different hand-arm postures, grip force (25–75 N) and vibration levels (27–53 mm/s_rms), were studied on eight subjects. The outcome clearly shows that the energy absorption properties of the human hand-arm system are more or less dependent on all of the experimental conditions studied, but mainly to the frequency of the vibration stimuli. Furthermore, the results indicate a non-linear relationship between the energy absorbed and all other variables studies.     

MAO-A inhibition in brain after dosing with esuprone, moclobemide and placebo in healthy volunteers: in vivo studies with positron emission tomography

Objective: The aim of the study was to investigate whether or not esuprone binds substantially to MAO-A in the human brain. Methods: In a randomised double-blind placebo-controlled study 16 male healthy volunteers were examined␣with positron emission tomography (PET) with [¹¹C]harmine. Eight of the volunteers were given daily doses of 800 mg esuprone, four were given bi-daily doses of 300 mg moclobemide, and four volunteers were given placebo tablets. PET was performed before initiation of a 7-day treatment period. On day 7, one investigation was made immediately before administration of the drug, representing 23 h after the previous day's treatment for esuprone and 11 h after the last tablets of moclobemide. Further investigations were made 4 h and 8 h after the morning dose on day 7. Results: PET showed a high degree of binding of [¹¹C]harmine, a high-affinity ligand for MAO-A, before the start of treatment, and a marked and similar reduction after treatment with esuprone and moclobemide. A slight tendency for normalisation of enzyme binding was observed at the last time point. In the placebo group no change was observed. Plasma kinetics of esuprone showed a rapid elimination with a half-life of about 4 h. Conclusion: The study demonstrates that esuprone was comparable to moclobemide in its effect on MAO-A inhibition in the brain at the doses given. This is an illustration of the potential of PET to monitor drug effects directly on target biochemical systems in the brain in human volunteers, and the possibility of using these data, rather than pharmacokinetic data, for the determination of dosing intervals. Received: 21 August 1996 / Accepted in revised form: 22 November 1996