Streptomyces rimosus extracellular proteases 3. Isolation and characterization of leucine aminopeptidase

Summary A leucine aminopeptidase was purified to homogeneity fromStreptomyces rimosus culture filtrates, which are waste broth of oxytetracycline bioproduction process. Purification procedure includes ultrafiltration and chromatography on CM-Sephadex, AH-Sepharose and FPLC Mono S column. The enzyme is a monomer with molecular weight of 27,500 Daltons and pI of 7.3, stable in broad pH range and up to 70°C. It is a metallo enzyme dependent on Ca²⁺ ions for its full activity. By its specificity it is a true aminopeptidase active on amino acid amide, arylamide, peptide and ester bonds. The hydrolysing activity shows preference for leucine at the N-terminal position of substrates, also acts on aromatic acids and methionine, but does not release glycine, proline, acidic amino acids orD-amino acid residues.     

Effect of the polysaccharide ofAgrobacterium radiobacter on the growth of plants and occurrence of damping-off in sugar beet

Agrobacterium radiobacter, strain B6 (a strain isolated in this laboratory, which limited the occurrence of damping-off of sugar beet and influenced growth of plants in hot-house and field experiments) was found to produce an acidic exopolysaccharide in a mineral medium with various carbon sources. Hydrolyzates of the polysaccharide contained glucose, galactose, glycerol, succinic acid and pyruvic acid, whose quantitative content varied according to the carbon source used. The polysaccharide isolated from the medium containing glucose exhibited the highest physiological activity. Seeds germinated best and sugar beet roots were found to grow most rapidly in a medium containing 0.2 % (W/W) of the polysaccharide. The roots exposed for 3 d in this medium grew 2.7-fold as compared with non-treated plants. Higher sumbers of microorganisms were detected on the surface of roots treated with the polysaccharide. Growth of roots was also stimulated when immersing the seeds (30 min) in a 0.2 –0.4 % solution of this polysaccharide. After a two-fold treatment the roots were less damaged by the fungusPythium ultimum. Plants from seeds treated with the polysaccharide grew in the field soil more rapidly than the non-treated plants but worse than after bacterization of the seeds byA. radiobacter B6 and were only partially protected against the damping-off of sugar beet.     

Interferon affects intracellular calmodulin levels

Interferon lowers calmodulin levels in two cell lines sensitive to its antiproliferative effect. Further, in synchronized cells, interferon strongly inhibits the increase in calmodulin observed when control cells enter the S phase, and concomitantly inhibits DNA synthesis. Calmodulin has been implicated in the control of cell proliferation and an increase in this protein seems to be necessary for the progression of cells into the S phase of the cell cycle. Therefore, the effect of interferon on calmodulin content might constitute part of the molecular mechanism by which interferon inhibits DNA synthesis.     

Preferential association of kappa IIIb light chains with monoclonal human IgM kappa autoantibodies

The predominance of the relatively uncommon V region subgroup isotype kappa III among the light chains of human monoclonal (IgM kappa) anti-IgG antibodies, (i.e., rheumatoid factors), was further documented through sequence analyses of ten such autoantibodies isolated from IgM-anti-IgG cold-insoluble immune complexes (mixed cryoglobulins). The amino-terminal sequence of all ten kappa-chains was characteristic for kappa III proteins and virtually identical to that of a prototype kappa III light chain. Similar sequence identity was found for kappa-chains isolated from three IgM kappa autoantibodies that formed cold-insoluble immune complexes with low-density lipoprotein (LDL). The thirteen light chains were found to be virtually identical in sequence for the first framework region (FR); ten of these proteins sequenced through the first complementarity-determining region (CDR) and into the second FR were markedly similar. The second CDR of five proteins was almost identical in sequence to that of the prototype kappa III-chain. Concordance was also demonstrated between the structural classification of the light chains as kappa III and their immunochemical classification as members of this V region subgroup. Serologic analyses of light chains isolated from seven IgM kappa autoantibodies (six anti-IgG, one anti-LDL) and of one intact IgM kappa anti-LDL antibody showed that each had antigenic determinants common to kappa II proteins. These light chains also expressed the antigenic determinant(s) of a V-region sub-subgroup of kappa III proteins designated kappa IIIb. Our studies confirm the preferential association of kappa III (and kappa IIIb) light chains with IgM kappa anti-IgG antibodies and demonstrate a similar association for IgM kappa anti-LDL antibodies. The finding that these and other types of IgM kappa autoantibodies, e.g., cold agglutinins, have remarkably similar light chains suggests an inherent restriction in the immune response to self-antigens.     

Restriction map of Tn7

Tn7, a transposon of 14 kb, encodes resistance to trimethoprim (Tp) and streptomycin (Sm). A cleavage site map of this transposon for twenty-two different restriction enzymes as determined by comparison of restriction enzyme cleavage patterns of the plasmids ColE1 and ColE1::Tn7 is presented. The precise localization of these sites was facilitated by the use of two deletion derivatives of ColE1::Tn7: pGB2 and ColE1::Tn7Δ6, and by the use of pOB14 and pOB15 which contain a part of Tn7 cloned into the plasmid pBR322. This map should aid in the study of the structural and genetic organization of this transposon.     

Recherches sur la conservation du virus de la densonucléose

Résumé Le virus de la densonucléose (VDN) conserve une partie de sa virulence en solution aqueuse entre 0°C et 30°C pendant deux mois. Son pouvoir infectieux se maintient à—10°C pendant deux mois mais est supprimé après dix jours de dessication. On peut done considérer ceParvovirus d’Insectes comme un virus relativement résistant aux températures que l’on rencontre ordinairement dans la nature et au laboratoire.

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Summary The persistence of DNV is described. This virus retains a part of its infectivity in aqueous solution between 0°C and 30°C. The log LD₅₀ initially 7,15 drops to 2 after two months. The virus keeps its infectivity at 10°C but it loses all its pathogenicity after 10 days in dry conditions.

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Ce travail a été réalisé en partie dans le cadre d’une convention I.N.R.A.-D.R.M.E.     

Regression analysis of the spontaneous spike activity of neurons in snail ganglia

Regression analysis of the spontaneous spike activity of neurons in Helix pomatia was carried out with the aim to establish the statistical parameters of this activity under constant experimental conditions and during longer time intervals. The activity of 38 randomly chosen neurons in visceral and parietal ganglia, penetrated by microelectrodes and activated either endogenously by pacemaker potentials or by synaptic inputs, was recorded during time intervals lasting from 20 min to 3 h. The main results of the statistical analyses are presented in the table where the parameters of both cell types are listed. The validity of the regression analysis applied here is discussed from the point of the possibility it offers for carrying on the data processing quickly and without applying complex calculating means. The results are also considered regarding the current interest of our research group.     

Struktur und quantitatives Verhalten des perimitochondrialen granulären endoplasmatischen Retikulums der Mäuseleber

Zusammenfassung In normalen Leberzellen der Maus wurde das quantitative Verhalten des perimitochondrialen granulären endoplasmatischen Retikulums untersucht. 74% der Mitochondrien zeigen Beziehungen zum granulären endoplasmatischen Retikulum. Die einzelnen Mitochondrien werden zu 52%19,5 von den Membranen des granulären endoplasmatischen Retikulums bedeckt. Je Mikrometer Membranstrecke sind auf der mitochondriennahen Seite des granulären endoplasmatischen Retikulums 21 und auf der mitochondrienfernen Seite 20 Ribosomen zu finden, was der Zahl im übrigen granulären endoplasmatischen Retikulum entspricht. Die Befunde stellen die Grundlage für Untersuchungen des perimitochondrialen granulären endoplasmatischen Retikulums unter pathologischen Bedingungen dar.

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Structure and quantitative behaviour of the perimitochondrial granular endoplasmic reticulum in the liver cells of the mouse

Summary The perimitochondrial granular endoplasmic reticulum in normal mouse liver cells has been investigated quantitatively. 74% of the mitochondria are in association with the granular endoplasmic reticulum. The individual mitochondrion is covered by the membranes of the granulated E. R. in 52%19.5. The outer surface of the endoplasmic membrane, facing the mitochondrion, is occupied by 21 ribosomes per m; the corresponding surface of the membrane facing the free cytoplasm is occupied by 20 ribosomes per m. These data are in agreement with those of that fraction of the E. R., which is not in association with mitochondria. These findings represent a basis for investigations of the perimitochondrial endoplasmic reticulum under pathological conditions.

     

Retting of Flax by Aspergillus niger

In this study, retting was carried out by Aspergillus niger. The pH, galacturonic acid (GA), and total reducing sugar were determined; the end point was identified by the classic empirical processes and by the maximal GA content of the retting water. The process gave clear and resistent fibers, and the retting time was similar to that of current industrial processes with bacterial enzymes. Control of total acidity was not required, since the pH remained close to neutrality throughout the entire process.     

Physiological properties of microbial population as a basis for graphical solution of multi-stage continuous cultivations

A proposal for a graphical solution of the design of a multi-stage continuous culture system is presented. The holding times in individual stages are derived from the reciprocal values of the growth rate and of the product formation rate and plotted against the cell and product concentrations. Characteristic changes of physiologically significant paramaters are then projected on these production curves. Dedicated to Academican Ivan Málek on the occasion of his 60th birthday