The endogenous lectins of the chick blastoderm are present in association with an apolipoprotein in distinct organelles and in the extracellular matrix

Summary Affinity purified preparations of the galactose-binding lectin from gastrulating chick blastoderms consist of three main polypeptides. Two of these have been identified as the 14 kD and 16 kD galactose-binding lectins. A third one migrates in SDS-PAGE gels with a relative molecular weight of 6,500±500 and has been identified as an apolipoprotein (Apo) of plasma very low density lipoproteins, Apo-VLDL-II. We have studied the localization of these polypeptides using immunofluorescence and ultrastructural immunocytochemistry with peroxidase and protein-A gold. The 14 kD lectin occurs in the intracellular yolk where it is mainly present within the electron lucent component. The 16 kD is also present in the intracellular yolk platelets, but tends to predominate in the electron-dense component. In addition, the 16 kD lectin is also present in pleiomorphic yolk-associated organelles and in the extracellular matrix. Apo-VLDL-II is also localized in the electron-lucent component of the yolk platelet and in the extracellular matrix. Our results suggest that the lectin(s) are associated with Apo-VLDL-II in the yolk platelet, and may subsequently become externalized.     

Identification of some lactic acid bacteria from two Zimbabwean fermented milk products

Ten predominant lactic acid bacterial isolates from traditionally fermented milk and four isolates from an industrially fermented milk, Lacto, together with 14 reference strains ofLactobacillus and three ofLactococcus were examined for 32 characteristics. Data were analysed using the simple matching coefficient and clustering was by unweighted pair group average linkage. All the isolated from traditionally fermented milk belonged to the genusLactobacillus. Seven isolates could be identified as belonging toL. helveticus, L. plantarum, L. delbrueckii subsp.lactis, L. casei subsp.casei andL. casel subsp.pseudoplantarum. Three of the isolates could only be identified as either betabacteria or streptobacteria. The four isolates from Lacto were identified asLactococcus lactis. However, they could not be identified to subspecies level.

---

Résumé On a examiné 32 caractéristiques de 10 souches de bactéries lactiques isolées de fait fermenté de manière traditionnelle, et de 4 souches isolées d'un lait fermenté industriel, le Lacto, ainsi que 14 souches de référence deLactobacillus et trols deLactococcus. Les données ont été analysées en utilisant le coefficient d'apparienient simple et le groupement par lien moyen de groupe pair non balancé. Toutes les souches isolées du lait fermenté de manfère traditionnelle appartiennent au genteLactobacillus. Sept souches ont pu étre identifiées comme appartenant àL. helveticus, L. plantarum, L. delbruckii subsp.lactis, L. casei subsp.casei etL. casei subsp.pseudoplantarum. Trois de ces souches n'ont pu être identifiées soit comme bétabactéries soit comme streptobactéries. Les quatres souches isolées de Lacto ont été identifiées commeLactococcus lactis. Celles-ci n'ont toutefois pas pu être identifiées jusqu'au stade de sous-espèce.

     

Subcellular Distribution of UDP-Galactose:Ceramide Galactosyltransferase in Rat Brain Oligodendroglia

Oligodendrocytes isolated from 18-19-day-old rat brain were homogenized in 0.32 M sucrose. The homogenate was centrifuged at 100,000 g for 50 min in a gradient containing 0.8, 1.05, and 1.3 M sucrose. Three discrete bands were obtained at the interfaces 0.32-0.8 (F1), 0.8-1.05 (F2), and 1.05-1.3 M (F3). The distribution of UDP-galactose:ceramide galactosyltransferase (CgalT) activity in each fraction was measured using liposomes containing normal fatty acid-containing ceramides (NFA-CgalT activity) or 2-hydroxy fatty acid-containing ceramides (HFA-CgalT activity). Although detection of both CgalT activities was possible in all fractions, HFA-CgalT activity was enriched in F1 and F2 fractions, which also showed an enrichment of Golgi and endoplasmic reticulum markers, respectively. It is interesting that NFA-CgalT activity was significantly enriched in the F2 fraction. These results suggest that hydroxylated and nonhydroxylated galactocerebrosides may be synthesized at different intracellular locations.     

What is Unique About Superoxide Toxicity as Compared to Other Biological Reductants? — A Hypothesis

Usually the toxicity of superoxide is attributed lo its ability to reduce metal ions and subsequently reoxidation of the metal by hydrogen peroxide yields deleterious oxidizing species. As many other nontoxic biological reductants reduce metal compounds, we suggest that part of the mechanism of superoxide toxicity results from its ability to oxidize metal ions bound to biological targets, which subsequently degrade the target via an intramolecular electron Transfer reaction.     

The distribution of A1 adenosine receptor and 5′-nucleotidase in pig brain cortex subcellular fractions

Pig brain cerebral cortex was subfractionated by isopycnic centrifugation in sucrose gradients. In each subfraction the content of the agonist [³H]R-PIA binding, the activity of adenosine metabolizing enzymes (5-nucleotidase and adenosine deaminase) and the activity of membrane marker enzymes were determined. The fractions were also examined by electron microscope. In general, the results suggest a widespread distribution of A₁ adenosine receptors in membranes from different origins. Marker enzyme profile characterization indicated an enrichment of A₁ adenosine receptor in pre-synaptic membranes isolated from the crude synaptosomal fraction (P2B subfraction) as well as in membranes of glial origin such as myelin. The receptor is also present in the endoplasmic reticulum and in membranes isolated from the microsomal fraction that seem to have a post-synaptic origin (P3B). In subfractions having a high content of adenosine receptor the equilibrium binding paramters were obtained as well as the proportion of high- to low-affinity sites. From the values of the equilibrium constants it was not possible to find differences between the receptor in the different subfractions. Analysis of the affinity state distribution showed a diminished percentage of high-affinity sites in fraction P3A, which can be accounted by the existence of myelin membranes; in contrast the percentage of high-affinity states was higher in P2 and P3B, indicating that in these fractions the receptor is present in synaptosomal membranes. The close correlation shown between the enzyme 5-nucleotidase specific activity and the specific ligand binding distributions led us to postulate an important role for the enzyme in the regulation of adenosine action in pig brain cortex.     

Cloning and characterization of an extracellular temperature-labile serine protease gene from Aeromonas hydrophila

Aeromonas virulence is thought to depend on multigenic functions. The gene for an extracellular protease from Aeromonas hydrophila SO2/2 was cloned in Escherichia coli C600-1 by using pIJ860, bifunctional plasmid, as a vector. The gene encodes for a temperature-labile serine protease (P2) with a molecular mass of approx. 68 kDa which is highly inhibited by PMSF. The gene was expressed in Streptomyces lividans 1326 by transforming protoplasts with the original clone pPA2. We were also able to transfer and express the prt P2 gene in Pseudomonas putida by mating experiments. The protein P2 was secreted into the periplasms of both P. putida and E. coli C600-1 being identical in properties to one of the proteases secreted into the culture supernatant by A. hydrophila SO2/2.     

Fruiting at High Temperature and Its Genetic Control in the Basidiomycete Flammulina velutipes

Of 10 geographic strains of Flammulina velutipes, 4 were found capable of fruiting at 22°C (Fr_H) rather than at the typical 15°C (Fr_L). Crosses made between Fr_H and Fr_L monokaryons were never observed to fruit at 22°C. However, some hybrids did fruit at the intermediate temperature of 18°C when grown on appropriate substrates, indicating incomplete dominance of the low-temperature requirement. Analysis of progeny of five Fr_H × Fr_L crosses indicated that a minimum of two genes appears to control the requirement for fruiting at ≤15°C. The genes are not closely linked to either incompatibility locus.     

On the transposition of copia-like nomadic elements in cultured Drosophila cells

We studied the stability of the genomic distribution of six retrotransposon families in long-term and short-term cultures of Drosophila cells. In a subclone derived from Kc cells, no significant rearrangements were detected over an 8 year period. On the contrary, extensive reshuffling and amplification of transposon families were observed in recently established cell lines. These results show that in cultured Drosophila cells transposition appears to be restricted to the transition from the embryo to continuous cell lines.     

Initial stages in the course of adventitious bud formation on embryos of Picea abies

Adventitious buds on embryos of Picea abies (L.) Karst. developed after a pulse treatment with 250 μ M benzyladenine (BA) of pH 5.5 for 2 h. Light and temperature regimes were not critical during the initial stages. Adventitious buds developed faster after a pulse treatment and the variation among different experiments was lower compared to when the embryos were cultured on media supplemented with BA. Various stages of the differentiation of adventitious buds were identified: stage 1 - appearance of meristematic centres (approximately the first two weeks); stage 2 - development of adventitious bud primordia (approximately the third week); stage 3 - adventitious bud development (from approximately the 4th to the 8th week). This system may be used for further studies on bud differentiation.     

Initiation and development of wound tissue and roots on hypocotyl cuttings of Pinus sylvestris in vitro

The rooting of hypocotyl cuttings from 20-day-old seedlings of Pinus sylvestris L. cultured in vitro is discussed. About 40% of the cuttings cultured on medium lacking activated charcoal produced roots during the first two months. When activated charcoal was added to the medium, either root formation (75% formed roots) or wound tissue growth (95% formed large wound tissues) was stimulated in different experiments. These large wound tissues did not develop any roots. The anatomical changes in the basal part of the cuttings were similar during the first two weeks in all the cuttings studied. A vascular cylinder composed of short tracheids with many pores developed. Thereafter the differentiation process became varied. The amount of wound tissue produced and the time for rooting differed among the cuttings. Tracheid nests which were in contact with the vascular system in the hypocotyl via short tracheids were observed after three weeks. Subsequently, roots developed from the tracheid nests. The longer root formation was delayed, the larger the wound tissue became.
Short tracheids were found close to the wound tissue surface. Their ability to adsorb nutrients and water is discussed.