Fibronectin and laminin in the extracellular matrix and basement membrane of sea urchin embryos

Fibronectin and laminin have been found in the extracellular matrix and in the basement membrane of sea urchin embryos during early development. These glycoproteins are also found on the cell surfaces of the outer epithelial layer and on the secondary mesenchyme cells within the blastocoel. The similarity of functions of the extracellular matrix and basement membrane is discussed, as is the similarity of their molecular components. These observations suggest the possibility that fibronectin and laminin form a continuous matrix surrounding the cells which links the outer ECM (hyaline layer) to the inner ECM (basement membrane). Such a network could coordinate the various activities of the embryo during early morphogenesis.     

Identification and characterization of gp TFA-1, a guinea pig T cell surface antigen associated with T cell function

Monoclonal antibodies (Ab) were produced that specifically recognized guinea pig T cells. FACS analysis revealed that Ab 188 bound to the majority of peripheral T lymphocytes of strain 2 and strain 13 guinea pigs and to a minor population of thymocytes. It failed to react with the Ia-bearing guinea pig B cell leukemia line EN-L2C, with macrophages, bone marrow cells, erythrocytes, or thrombocytes. Treatment of T cells with Ab 188 and complement prevented T cell activation. Culturing primed T cells with antigen- or mitogen-pulsed syngeneic or with allogeneic macrophages in the continuous presence of Ab 188 produced a marked, dose-dependent inhibition of T cell proliferation. The antigen defined by Ab 188 was therefore designated guinea pig T lymphocyte function-associated antigen-1, gp TFA-1. The magnitude of inhibition by Ab 188 varied between 65 and 85% whereas three other antibodies to guinea pig T cells had no inhibitory effect on T cell proliferation. Time course experiments revealed that gp TFA-1 is critically involved in an early phase of T cell activation. Maximal inhibition was achieved only if the antibody was present from the beginning of the cell culture; the addition of antibody after 24 hr of culture no longer had an inhibitory effect. Ab 188 did not induce T cell mitogenesis. Two-dimensional analysis (one-dimensional, IEF; two-dimensional, SDS-PAGE) of immunoprecipitates obtained from NP40 lysates of [35S]methionine-labeled T cell blasts indicated that a molecule was specifically precipitated that consisted of two noncovalently associated polypeptide chains with apparent m.w. of 43,000 and 38,000. Both subunits displayed extensive charge heterogeneity focusing at an average isoelectric point of 5.0 and 6.5, respectively. The gp TFA-1 molecule exhibits striking similarities in its functional and structural properties to recently described clonotypically expressed T cell glycoproteins, which were shown to be involved in antigen recognition by T cells in the murine and human systems.     

Serum Albumin Washing Specifically Enhances Arachidonate Incorporation into Synaptosomal Phosphatidylinositols

Arachidonate incorporation into synaptosomal phospholipids was shown to be affected by factors including the procedure for preparation of the membrane fractions and preincubation of synaptosomes prior to assay of incorporation of arachidonate into both phosphatidylcholine (PC) and phosphatidylinositol (PI). However, the inhibition toward incorporation into PIs, but not PCs, was fully reversed when the membranes were washed with bovine serum albumin. A twofold increase in arachidonate incorporation into PIs was also observed when freshly prepared synaptosomes were washed with serum albumin immediately before assay of incorporation activity. The inhibitory action is thought to be due to an increase in polyunsaturated fatty acids and/or their oxidation products which may then elicit a special effect on the acyltransferase responsible for transferring arachidonate into phosphatidylinositols. The differences in fatty acid uptake and response to serum albumin also suggest the presence of different acyltransferase for acyl transfer to PIs and PCs.     

Experience using two CT-guided stereotactic biopsy methods

15 patients had intracranial CT-guided stereotactic biopsies. Biopsies were performed either with a Riechert-Mundinger stereotactic frame modified for use in the CT or by using the CT scan to establish the relationship of the intracranial lesion to identifiable bony landmarks, and subsequently performing the biopsy in a standard stereotactic frame. Both systems provided safe and accurate methods for obtaining intracranial tissue.     

Wirtskreis und Infektionscyclus eines neu isolierten Bdellovibrio bacteriovorus-Stammes

Zusammenfassung Der neu isolierte Stamm W von Bdellovibio bacteriovorus infiziert und lysiert Rhodospirillum rubrum F und alle anderen untersuchten Athiorhodaceae, nicht aber Pseudomonas aeruginosa und Spirillum serpens. Er befällt auch zahlreiche Enterobacteriaceae und von den grampositiven Bakterien Streptococcus faecalis und Lactobacillus plantarum.Nach dem Festheften an der Zellwand wird diese in 3–20 min durchdrungen. In 10–60 min ist Bdellovibrio vollständig in die Zelle eingedrungen und hat sich im Raum zwischen Zellwand und cytoplasmatischer Membran angesiedelt.In 3–5 Std wird der gesamte Zellinhalt bis auf die Membranen aufgelöst. In dieser Phase erfolgt die Vermehrung von Bdellovibrio. In den ghosts sind die Parasiten in lebhafter Bewegung. Die Geißel hat einen Gesamtdurchmesser von 29 m und eine Länge von etwa 3 . Sie ist von einer Geißelscheide umgeben, die in Verbindung zur Zellwand steht. Der Durchmesser der Geißel ohne Scheide beträgt etwa 18 m. Bdellovibrio kann oberhalb eines Sauerstoffpartialdruckes von 4–5 mm Hg infizieren und sich vermehren. Der Titer von Bdellovibrio nimmt bei Aufbewahrung in lysierten Kulturen in 36 Tagen von 10⁸ auf 10¹ pfu (plaque forming units) je ml ab. Bei Aufbewahrung in Nährkultur sinkt der Titer nur auf 10⁴ pfu/ml ab. Die Zahl der Plaques im Verhältnis zum Titer der Impfsuspension von Bdellovibrio schwankt in Abhängigkeit vom Wirtsstamm. Wenn man die Plaque-Bildungsrate bei R. rubrum gleich 1 setzt, beträgt sie bei Serratia marcescens 0,0001, bei Proteus vulgaris 10. Bd. bacteriovorus, Stamm W wächst nicht in synthetischer Nährlösung oder Lysaten. Ein geringes Wachstum ohne Zellteilung findet in Zellextrakten von R. rubrum statt. Der Stamm vermehrt sich jedoch in hitzeinaktiviertem R. rubrum. Die Plaque-Bildungsrate ist unter diesen Bedingungen aber sehr niedrig.In Lysaten treten encystierte Dauerformen von Bdellovibrio bacteriovorus auf.

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The host range and the infectious cycle of a new isolated, on gram-positive and gram-negative bacteria parasiting Bdellovibrio bacteriovorus strain

Summary Rhodospirillum rubrum and all other investigated Athiorhodaceae are infected and lysed by the new isolated strain W of Bdellovibrio bacteriovorus. This strain W parasites on numerous Enterobacteriaceae and the gram-positive bacteria Streptococcus faecalis and Lactobacillus plantarum, but not on Pseudomonas aeruginosa and Spirillum serpens.After attachment of Bdellovibrio to the host, the cell wall is penetrated in 3 to 20 min. In 10 to 60 min Bdellovibrio has completely entered the host cell. He remains in the space between cell wall and cytoplasmic membrane of the host.The host cell is completely lysed within 3 to 5 hours. During this phase the size and cell number of Bdellovibrio are increased and a new flagellum is likely to be formed. In the ghosts of the host cell a strong movement is observed. The single polar flagellum of Bdellovibrio has a diameter of 29 m. The flagellum consists of an inner core ( 18 m) and an outer sheath which is continued into the cell wall. Bdellovibrio is able to grow and to infect only under aerobic or semiaerobic conditions (oxygen partial pressure 4 to 5 mm Hg and more). The titer of Bdellovibrio is gradually decreased from 10⁸ to 10¹ plaque forming units (pfu) per ml, when kept in the lysate for 36 days. In a synthetic medium there is a diminution of 10⁴ pfu/ml only. The plating efficiency is dependent of the host strain. If the plating efficiency of Bdellovibrio with Rhodospirillum rubrum is 1.0, the rate varies from 0.0001 with Serratia marcescens to 10 with Proteus vulgaris. Bdellovibrio bacteriovorus strain W does not grow in a synthetic medium. However, it grows but does not multiply in cell free extracts of Rhodospirillum rubrum. The parasite is also able to infect and lyse heat inactivated R. rubrum. But the plating efficiency in this case is very low.It has been observed that in lysed cells of R. rubrum certain amount of Bdellovibrio is encysted. The morphology and fine structure of these cells is quite different from the normal virulent type.

     

The Distribution of Acidic and Neutral Peptidases in Barley and Wheat Kernels

The activity of barley and wheat peptidases which hydrolyze alpha-N-benzoyl-dl-arginine-p-nitronnilide (BAPA) and alpha-N-benzoyl-l-arginine ethyl ester (BAEE) has been measured in proximal and distal portions of ungerminated grain and in these tissues during 6 and 7 day incubations. The proximal portion of ungerminated barleys contained the major part of both the acidic (BAPA-ase and acidic BAEE-ase) and neutral (neutral BAEE-ase) peptidases. In ungcrminated wheat these acidic and neutral peptidases were nearly evenly distributed between the proximal and dislal portions. Commercial wheat embryo was very high in acidic peptidase but contained no neutral peptidase. During the germination of both wheat and barleys, acidic and neutral peptidase activity in the seedlings increased with time. No such consistent increase was observed for aleurone and starchy endosperm tissue for any of these grains. Aleurone and starchy endosperm tissue incubated in the absence of the proximal portion of the kernel showed reduced peptidase activities.