Adaptive control of intracellular Ca2+ release in C2C12 mouse myotubes

Laser scanning confocal imaging was used to monitor release of Ca²⁺ from localized regions in a skeletal muscle cell line with sparsely distributed Ca²⁺ release sites. The goal was to distinguish between two schemes proposed to explain the phenomenon of “quantal” Ca²⁺ release from caffeine-sensitive Ca²⁺ stores in muscle and other tissues: (1) all-or-none (true quantal) Ca²⁺ release from functionally discrete stores that have different sensitivities to caffeine; or (2) adaptive behavior of individual release sites, each responding transiently and repeatedly to incremental caffeine doses. Our results showed that Ca²⁺ release induced by K⁺ or caffeine occurs in discrete loci within the cell. The image areas and fluorescence intensities of some of these evoked local signals were similar to those of Ca²⁺ sparks that were observed under resting conditions and which are believed to be due to spontaneous activation of single release units. In contrast to the expectations imposed by quantal models, incremental doses of caffeine activated the same sets of release sites throughout the cell. Ca²⁺ release, at a given site, triggered by a submaximal dose of caffeine was transient and could be reactivated by addition of a higher caffeine dose, showing the same type of adaptive behavior as measured globally from larger areas of the cell. These results suggest that incremental Ca²⁺ release is accounted for by adaptive behavior of individual Ca²⁺ release sites. Received: 9 August 1995/Received in revised form and accepted: 13 October 1995     

Diagnosis of penicillin allergy by means of Phadebas RAST penicilloyl G and V and skin tests

Fifty patients with suspected allergy to penicillin were tested. Skin tests were done with Na-penicillin G and penicilloyl-polylysin. Specific IgE antibody assays were done with penicilloyl G and V conjugates by means of RAST. The overall agreement between skin test and RAST results was 87%, borderline cases not included. In one case, skin tests were positive to penicillamine only, while RAST for penicilloyl G and V both proved to be positive. One case of penicillin allergy could be diagnosed in vitro post mortem only. Two cases of Hoigné syndrome showed no evidence of allergy. Patterns of skin manifestations varied but urticaria was the most commonly seen feature. Twenty patients without adverse reactions to penicillin treatment and seven patients who had not received penicillin over the last 10 years served as controls. None of them were positive in either skin tests of RAST. Two or our twenty control patients developed penicillin allergy during the study. Both showed positive RAST results.     

MIF in der therapiekontrolle von herpes simplex recidivans: Behandlung mit levamisole,BCG, urushiol und herpes antigen vaccine

Zusammenfassung Dreißig Patienten mit rezidivierendem Herpes simplex und 6 Kontrollpatienten wurden in unserer Studie erfaßt. Vor der Behandlung, sowie während und nach der Therapie mit Levamisole, BCG, Urushiol und Herpes Antigen Vaccine wurden MIF-Bestimmungen durchgeführt.Dabei erwies sich der MIF als wertvoller Test in der Diagnose der Bereitschaft Herpesrezidive zu bekommen und in der Beurteilung des Therapieerfolges.Die Behandlung mit Levamisole, BCG und Herpes Antigen Vaccine war erfolgreich.Dabei korrelierten die MIF-Bestimmungen mit dem Ergebnis der Levamisole-und Herpes Antigen Vaccine-Behandlung, nicht hingegen bei den mit BCG behandelten Patienten.

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Therapy of recurrent herpes simplex and its surveillance by MIF determination: with levamisole, BCG, urushiol and herpes antigen vaccine

Summary Thirty patients with recurrent herpes simplex and 6 controls were included in our study. Before initiation of treatment, during therapy with Levamisole, BCG, poison ivy and herpes antigen vaccine and, thereafter, MIF-determinations were performed.The latter test proved to be a useful parameter in evaluating the sensitivity against herpes infection, (and subsequent recurrent manifestations), and effect of therapy.Treatment with Levamisole, BCG and herpes antigen vaccine was successful. MIF-inhibition values paralleled the therapeutic effect in the Levamisole- and herpes antigen vaccine treated group, not however, in the BCG-treated patients.

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Auszugsweise vorgetragen auf der IV. Jahrestagung der Arbeitsgemeinschaft für Dermatologische Forschung, 22.–24. Oktober 1976 in Berlin     

Preparation,measurement and possible use of human antitoxin againstCl. botulinum A,B, and E toxins

Human antibodies against botulinum toxins A, B, and E have been raised by repeated injections of pentavalent toxoid in a healthy volunteer. The final titer was 3.2 U anti-A, 0.4 U anti-B, and 2.5 U anti-E/ml. In mice, the efficacy of the antitoxin decreases with the time between poisoning and application of the antiserum. The dose recommended for prophylactic purposes in man is 1 ml/kg. In overt poisoning, therapy should be started with higher titer animal serum since in animal experiments high titer sera can stop (although not reverse) the symptoms of overt botulism within an-admittedly not too broad-rnage of time and dosage. Later on, therapy can be continued with human antiserum. An inverse radioimmunassay for botulinum A antitoxin using labeled botulinum toxin and antibody-coated tubes is described. The serum is available upon request from the author.     

Autologous olfactory glial cell transplantation is reliable and safe in naturally occurring canine spinal cord injury

Intraspinal transplantation of olfactory glial cells (OGC) has produced well-defined beneficial effects in experimental rodent models of spinal cord injury (SCI) and therefore has considerable promise as a treatment for severe SCI in human patients. In this study, we used clinical canine cases of severe SCI to determine whether derivation and transplantation of OGC from an autologous source was feasible. From the nerve fiber layer of a single olfactory bulb, we were able to generate 5 x 10(6) cells from each patient within 3 weeks. Of this population, 72% were p75(+) OGC, 20% were meningeal cells, and the remainder mainly astrocytes. Intraspinal transplantation was not associated with any observable long- or short-term complications.     

Traumatic superior mesenteric arteriovenous fistula and aortic pseudoaneurysm 20 years after repair

A case of traumatic superior mesenteric arteriovenous fistula (SMAVF) and aortic pseudoaneurysm successfully treated by a unique combination of operative and endovascular techniques with a 20-year follow-up is reviewed. After 20 years, the patient presented with an aortoenteric fistula, which was managed with a cryopreserved aortic interposition graft. In this report, we review the evolution of the treatment for traumatic SMAVF and aortic pseudoaneurysm and the current management of aortoenteric fistula.     

Donor hematopoietic cells from transgenic mice that express GFP are immunogenic in immunocompetent recipients

OBJECTIVE: To study the immunogenicity of hematopoietic cells marked with green fluorescence protein (GFP) while avoiding the potentially confounding effects of viral gene transduction, marked cells from GFP+ transgenic mice were tracked after transplantation into unconditioned immunocompetent recipients. MATERIALS AND METHODS: Marrow was harvested from GFP+ transgenic mice that had been crossed onto a BALB/cByJ background. Unconditioned marrow transplantation involved infusion of sex-matched or sex-mismatched cells into female BALB/cByJ hosts. Engraftment and contribution to circulating nucleated blood cells were compared to recipients of donor cells that were not GFP-marked. Donor cells were detected by flow cytometry (GFP) and fluorescence in situ hybridization (FISH) for Y-chromosome sequences. RESULTS: Donor cells from mice of the same genetic background that did not express GFP were detected for more than four-weeks in unconditioned recipients. In contrast, GFP-marked cells in the blood peaked at one-week, declined to undetectable levels by two-weeks and were not detected in the marrow at sacrifice. In sex-mismatched studies, detection of male GFP+ donor cells by FISH yielded levels similar to those observed by flow cytometry, in contrast to the levels detected for many weeks in mice infused with male cells that did not express GFP. In immunocompetent recipients immunized with irradiated GFP-expressing cells, rechallenge with GFP+ cells resulted in the accelerated loss of donor cells. CONCLUSION: Donor marrow cells from GFP+ transgenic mice were lost after infusion into unconditioned immunocompetent mice and sensitization studies infer an immunologic mechanism. These results are similar to studies of virally transduced cells. Thus, infusion of cells with optimum engraftment potential could not compensate for the loss of donor cells due to immunogenicity.