C. Mittermayer

Lethal Complications of Typhoid-Cholera-Vaccination (Case Report and Review of the Literature)

Simultaneous parenteral vaccination against typhoid and cholera lead to death through either anaphylactic shock or endotoxic shock in a 36-year-old male. At autopsy the charactertic features of shock as well as chronic interstitial myocarditis were noted. Moreover, fresh histiocytic and lymphocytic nodules were found in the liver, heart and meninges. A review of the literature dealing with lethal complications following parenteral tyhoid vaccinations shows an increased risk in debilitated persons (emaciation, stress, cold). Most of the fatalities occurred in persons who had previous disturbances of the cardiovascular system, as in the case reviewed here. Cardiac failure, Landrys paralysis, renal failure and disturbances of skin, joints and intestines may also follow typhoid vaccinations. However, these latter complications are usually not lethal. The patients presented here had many of the conditions which are known to aggravate the situation and to lead to a lethal culmination. The review of this case and the disucussion following it shows that only healthy persons should receive the parenteral typhoid vaccination. Hopefully, the presentation of this material will help prevent fatalities of this type in the future.

DNA synthesis during the life cycle of L cells: Morphological, histochemical and biochemical investigations with arabinosylcytosine and thioarabinosylcytosine☆

Abstract DNA-synthesis during the life cycle of L cells: Morphological, histochemical and biochemical investigations with ARA-C and thio-ARA-C. Both ARA-C and thio-ARA-C effect a rapid decrease in the mitotic index of L-cells. At the same time giant cells are formed. Volumetric and planimetric measurements show that this is due to equal accumulation of substances in the cyoplasm and in the nucleus. Cytophotometrically the effects of both cytostatics are characterized by selective killing of S and accumulation of G 1 -phase cells. Also a small accumulation of G 2 -phase cells is seen, suggesting a block between G 2 and mitosis. The giant cells are charaterized by their DNA content as G 1 -phase cells but contain increased masses of RNA (25%) and proteins (300%) . It is shown that one of the enzymes involved indeoxycytidine metabolism (dCMP-deaminase) is greatly enhanced by ARA-C both in randomly and synchronously dividing cells. Simultaneously a shift from soluble to particulate conformation is observed in DNA-polymerase activities. On the basis of these observations combined with earlier investigations on thymidine-kinase and CDP-reductase, a theory for the action of high and low concentrations of ARA-C and thio-ARA-C is proposed.

Experimental Investigations into the Pathogenesis of Endocarditis due to Shock

Generalized Shwartzman reaction (GSR) was induced in rabbits by two intravenous injections of endotoxin 24 hours apart. Consumption coagulopathy was found in all cases. Histological investigations showed an abacterial endocarditis (thrombotica or verruccosa simplex), comparable to the one seen in humans after shock, in 69% of the treated animals. Biopsies of mitral valve were investigated by electron microscopy. In the early stage of GSR the valvular alterations were characterized by an oedema of the entire valve, an increase in the number of microvesicles in the endothelial cytoplasm, and a marked enlargement of endothelial cell surface by numerous cytoplasmic projections. At a later stage autophagic vacuoles, often showing a destruction of their membranes, and degenerative changes in cytoplasmic organelles occurred in many cells of the valve. On the other hand there were cells in the endothelium, which presented a distinctly increased number of Golgi complexes and of cisternae of rough endoplasmic reticulum. In other regions ruptures of endothelial plasmalemma, as well as hemorrhages into the valvular stroma were observed. Thus predestined loci for thrombogenesis were formed. In conjunction with these structural changes three factors may be involved in the formation of endocarditis due to shock: 1. hypercoagulability in the course of consumption coagulopathy, leading to precipitation of fibrin and platelets in the circulating blood; 2. turbulences of the blood flow, which are produced by the closure of the heart valves, and favour thrombogenesis on valves: 3. direct changes of the endocardium during endotoxemia. Quantitative changes in the different compartments of intracytoplasmic organelles in many cells of the endothelium during the later stage of GSR are discussed.

Bleomycin: Wirkung auf DNS und zellkernstrukturen in mäusefibroblasten

SummaryIn asynchronously dividing mouse fibroblasts (L-cells) the antibiotic Bleomycin exerts a cytostatic effect by selectively inhibiting DNA-synthesis. Our experiments show scissions in DNA of double and single strands in vivo, leading to small, partly acid soluble DNA pieces. Only after fragmentation of DNA the cells accumulate in G2-phase of the cell cycle. Simultaneously, alterations in the nuclear structure occur. First, the electron-density of eu- and heterochromatic regions decreases, resulting in an over-all loss of chromatin mass. Later, also nucleolar structures are transformed into “nucleolar caps”. Apparently, the fragmentation of DNA leads to a destabilization and denaturation of DNA and consequently to conformational changes of chromatin structure.ZusammenfassungAn asynchron wachsenden Mäusefibroblasten (L-Zellen) wurden die Effekte von Bleomycin auf die DNS, die phasenspezifische antimitotische Wirkung im Zellcyclus sowie die ultrastrukturellen Veränderungen am Zellkern untersucht.Unsere Experimente zeigen das Auftreten von Einzel- und Doppelstrangbrüchen der DNS bei Bleomycin behandelten Zellen in vitro. Die celluläre DNS wird dadurch in kleine, teils säurelösliche Fragmente zerlegt. Die behandelten Zellen werden phasenspezifisch in der G2-Phase des Zellcyclus arretiert. Dieser Arrest ist erstnach Fragmentierung der DNS wirksam. Gleichzeitig verändert sich die Zellkernstruktur. In einem ersten Stadium vermindert sich die Elektronendichte sowohl in eu- wie heterochromatischen Bezirken. Die Gesamtmasse des Chromatins nimmt ab. Später beobachtet man eine Segregation der nucleolären Bestandteile zu “nucleolar caps”.Offenbar führt die Fragmentierung der DNS unter Bleomycin zu einer Destabilisierung und Denaturierung der DNS mit nachfolgender Konformationsänderung der komplexen Chromatinstrukturen.

Intravital microscopy and histology of the generalized Shwartzman reaction in rabbits passively tolerant to endotoxin.

Summary The generalized Shwartzman reaction was significantly inhibited in rabbits pretreated with serum of endotoxin tolerant rabbits. Whereas coagulation studies revealed only moderate decrease of platelets, intravital microscopical and histological examinations showed even in tolerant animals a diffuse vascular lesion, but no fibrin thrombi and no parenchymal necrosis. It was concluded that endotoxin is directly toxic to vascular endothelium and that the vascular lesion in the generalized Shwartzman reaction is not mediated primarily by disseminated intravascular coagulation.

Effect of bromocarbamide on fibroblasts in vitro: fine structural changes in mouse fibroblasts.

Summary As part of our research series with bromocarbamide, we examined the ultrastructural effect of bromocarbamide on cultured fibroblasts. Monolayers of the cells were grown on millipore filters in the presence of dissolved bromocarbamide and the filters were embedded in capsules so that longitudinal cross-sections of the cells could be observed. Electron microscopically, we observed increased numbers of microvilli and increased pseudopodia at the external cell surface, increased pinocytotic activity (examined with exogenous peroxidase), increased lipid droplet aggregation in the cytoplasm and increased cellular overlap. Two possibilities are proposed to explain these phenomena: one involving direct attack of bromocarbamide on the plasma membrane, and the other, a direct interference with cellular metabolism.

Effect of bleomycin on the fine structure of mouse fibroblasts.

Asynchronously dividing mouse fibroblasts (L-cells) treated with the antitumor antibiotic Bleomycin show various rather specific morphological alterations. Many of the cells exposed to bleomycin assume a more or less epitheloid cell shape and are larger than untreated cells; in addition to an increase in nuclear size these cells often contain multiple nuclei. In most of the cells nuclei show an almost complete loss of peripheral condensed chromatin. The nucleolar hypertrophy initially observed is followed by a shrinkage and segregation of the nucleolar components. The cytoplasmic alterations include dilatation of the cisternae of the rough endoplasmic reticulum as well as an increase of free, non membrane attached ribosomes, often arranged in spiral- and rosette-shaped polysemes; they are not specific for bleomycin. Asynchron wachsende Mäusefibroblasten (L-Zellen) ändern unter der Einwirkung von Bleomycin ihre typische Zellform. In den vergrößerten, teilweise epitheloiden Zellen finden sich häufig multiple Zellkerne; die Nucleolen sind vergrößert. Eine langdauernde Bleomycin-Behandlung führt hingegen zu einer Verkleinerung der Nucleolen, die häufig mit einer Segregation der nucleolären Bausteine einhergeht. Die auffallendsten ultrastrukturellen Veränderungen bestehen in einem nahezu vollständigen Verlust besonders des kondensierten randständigen Kernchromatins, in einer starken Erweiterung der Cysternen des rauhen endoplasmatischen Reticulums sowie in einer Zunahme der freien, nicht membrangebunden Ribosomen.

Characterization of a Nutritive Synchrony in L-cells

Summary A synchronization method in L-cells producing nutritive stagnation of the cells in G 1 is presented. After reinnoculation, cells proceed through the cycle in the normal 24 hours. The method has the advantage that no chemical blockers are added to the system and a large number of cells is obtained. In a comparison with other synchronizing systems, this system rates in the foreground. Desynchronization due to individual cell physiologies makes investigations of G 2 difficult with this system. The method is uncomplicated and probably applicable to similar cell types.

Inhibition of the generalized Sanarelli-Shwartzman reaction by passive induction of endotoxin tolerance

The generalized Sanarelli-Shwartzman reaction (GSSR) and the known decrease of platelets and of the coagulation factors I, V, VII and VIII after the second injection of lipopolysaccharid was significantly inhibited in rabbits pretreated with serum from endotoxin-tolerant rabbits. Maximal protection against the GSSR and the decrease of platelets was conferred by serum fractions of tolerant animals obtained by gelfiltration on Sephadex G-200, containing the macromolecular fraction I.SummaryThe generalized Sanarelli-Shwartzman reaction (GSSR) and the known decrease of platelets and of the coagulation factors I, V, VII and VIII after the second injection of lipopolysaccharid was significantly inhibited in rabbits pretreated with serum from endotoxin-tolerant rabbits. Maximal protection against the GSSR and the decrease of platelets was conferred by serum fractions of tolerant animals obtained by gelfiltration on Sephadex G-200, containing the macromolecular fraction I.ZusammenfassungDie vorliegenden Untersuchungen zeigen, daß eine Übertragung von Serum endotoxintoleranter Kaninchen vor der präparativen Endotoxininjektion bei den Empfängertieren signifikant die Entwicklung eines generalisierten Sanarelli-Shwartzman Phänomens und den nach der zweiten Lipopolysaccharidinjektion einsetzenden Abfall der Thrombocyten und der Gerinnungsfaktoren I, V, VII und VIII hemmt. Ebenso schützt eine Übertragung der durch Gelfiltration auf Sephadex G 200 aus dem Serum endotoxintoleranter Kaninchen abgetrennten großmolekularen Fraktion I die Empfängertiere vor der Entwicklung eines generalisierten SSP und vor dem nach der zweiten LPS-Injektion einsetzenden Thrombocytenabfall.

Bleomycin: effect on satellite DNA in mouse fibroblasts

The effect of Bleomycin on the semiconservative replication of mouse nuclear DNA has been studied. When asynchronously dividing mouse fibroblasts (L-cells) were grown in the presence of 5-bromodeoxyuridine (25mg/l medium) for 18h, three hybrid DNA bands with densities of 1.722, 1.752, and 1.761 kg/l appeared after caesium chloride density gradient centrifugation of nuclear DNA. In cells exposed to Bleomycin (100 mg/l) however, replication of satellite DNA is more strongly inhibited than is the replication of the main band DNA; preferentially the thymidine-rich hybrid duplex at 1.761 kg/l could no longer be detected. Der Einfluß von Bleomycin auf die semikonservative Reduplikation der Kern-DNS von Mäusefibroblasten wird untersucht. Nach 18 stündiger Inkubation asynchron sich teilender L-Zellen mit Bromdesoxyuridin (25 mg/l Nährmedium) trennt sich die Kern-DNS in der Cäsiumchlorid-Dichtegradientenanalyse in 3 hybride Banden mit den Dichten 1,722, 1,752 und 1,761 kg/l. Unter der Einwirkung von Bleomycin (100 mg/l) wird die Reduplikation der Satelliten-DNS, im besonderen die des thymidin-reichen Stranges, weit stärker gehemmt als die der Hauptbanden-DNS.