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Anti-influenza virus activity of a lignin fraction from cone of Pinus parviflora Sieb. et Zucc.

When mice were inoculated intranasally or intracerebrally with lethal doses of influenza virus A/WSN/33, most died within 12 days. However, the infectivity of virus that had been preincubated with a lignin prepared from cones of Pinus parviflora Sieb. et Zucc. (PC-Fr. VI) was significantly reduced. Intraperitoneal or oral administration of PC-Fr. VI, prior to virus inoculation, slightly increased the survival ratio of the infected mice. Experiments using radiolabeled PC-Fr. VI revealed that this fraction effectively binds to virions as well as to cultured cells. These data suggest that PC-Fr. VI either inactivates the virus or induces the anti-viral state in cells by binding to virions or cells.

Administration, Oral↗

[Competition of isogenic haploid and diploid cells of the yeast Saccharomyces cerevisiae and Pichia pinus growing in mixed populations].

During "quasi-continuous" cultivation in rich and minimal media diploid yeast cells of Saccharomyces cerevisiae completely displace isogenic haploid ones. When Pichia pinus are cultivated in the minimal medium, the diploids also have an advantage over isogenic haploids. The results are discussed within the framework of the hypothesis of fixation of diploid phase in the course of biological evolution.

Diploidy↗

Antimicrobial spectrum of lignin-related pine cone extracts of Pinus parviflora Sieb. et Zucc.

Pine cones of Pinus parviflora Sieb., et Zucc, were extracted successively with 5% NaHCO3, 3% NH4OH, 1% NaOH and 4% NaOH, and the extracts were tested for ability to induce antimicrobial activity in mice infected with Staphylococcal aureus. Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Candida albicans, or Salmonella enteritidis. These alkaline extracts were potent against the first 5 of these strains of microorganisms, and the activity was recovered mostly from their acid-precipitates at pH 5. After further fractionation on a Sephadex LH-60 column, the highest molecular weight fraction was most potent. Chemical structures in these bioactive fractions were assumed to be lignin-related structures, based on spectral data from UV, NMR and IR spectroscopy.

Anti-Bacterial Agents↗

[Systemic allergic reaction after ingestion of pine nuts, Pinus pinea].

An in vivo open oral provocation with pine nuts (Pinus pinea) confirmed information about systemic reaction after ingestion of pine nuts. In vitro tests suggested a systemic IgE allergic reaction. Pine nuts are employed in sweets and cakes and, as in the present case, in green salads.

Adult↗

["Illegal" transformation of red adenine-dependent mutants of the yeast Pichia pinus by the pYE(ADE2)2 plasmid].

The red adenine-dependent mutants ade1 of the yeast Pichia pinus blocked in the VI step of adenine biosynthesis (lack of AIR-carboxylase) and ade2 mutants blocked in the VII step of adenine biosynthesis (lack of SAIKAR-synthase) were transformed with the plasmid pYE(ADE2)2 containing ADE2 gene of Saccharomyces cerevisiae encoding AIR-carboxylase. The appearance of white Ade+ clones with the frequency 2-7.10(-8) (which is ten-fold higher than reversion frequency) was only observed in the case of ade2 transformation. Genetic analysis points to connection of the "illegitimate" transformants' appearance with the change in the mutant ade2 locus or in a locus closely linked to the former. Ade+ phenotype was stable during 20 generations of mitotic budding. Southern blotting assay of transformant chromosomal DNA indicates that reconstitution of ade2 defective gene is related with its "correction", owing to integration of pYE(ADE2)2 sequence in the vicinity of the mutant locus.

Adenine↗

Antitumor activity of polysaccharide fractions from pine cone extract of Pinus parviflora Sieb. et Zucc.

Hot water extract of pine cone (PCE) of Pinus parviflora Sieb. et Zucc. dose-dependently suppressed both solid and ascites tumor cells transplanted into various mice. Acidic polysaccharides of PCE significantly increased the survival time of mice bearing ascites tumor cells, and activity increased with acidity. One of the four polysaccharide fractions obtained by NaOH extraction showed the most potent antitumor activity. This fraction significantly suppressed the growth of solid tumor cells, with occasional tumor regression and necrosis, and with little or no cytocidal effect on cultured tumor cells. All acidic polysaccharides were able to activate mouse macrophage-like cell line J774.1. There did not appear to be any correlation between the antitumor activity of these polysaccharides and their content of arabinose (or fucose), mannose, galactose, glucose, or uronic acid.

Animals↗

[Patterns in the cellular organization of ovule growth in the pine, Pinus silvestris L. I. Growth processes in endosperm].

The dynamics of growth processes was studied in the endosperm of Pinus silvestris during the year of fertilization. The growth of endosperm is described by an diauxic curve where the period of delay corresponds, by time, to fertilization. The growth of tissue proceeds, mainly, at the expense of cell proliferation. The mitotic activity is preserved in the endosperm till the beginning of formation of reserve substances in the cells; cell division and elongation occur simultaneously. The unfavourable environmental conditions influence the mitotic activity of cells only. The size of fully grown cells is relatively constant and somewhat decreases later, during maturation. One can distinguish rather early those regions in the endosperm where the cell size is 2-3 times less than in the rest tissue; the size of cells and nuclei in these regions suffers only insignificant changes during the growth. The large nuclei are observed in the beginning of the cell endosperm formation and, then, their size decreases 1.5-2 times; the second increase in nuclear size in the endosperm is observed during the period prior to the formation of reserve substances.

Animals↗

Biochemical changes in Pinus pinea seeds during storing.

The changes in the germination-rate, the contents in germination-inhibitors and the biochemical differences in soluble proteins and nucleic acids in freshly harvested Pinus pinea seeds stored for various periods of time, up to 24 months, and at two different temperatures (room temperature and 4 degrees C), have been investigated. The present results show that the maturation or after-ripening process of this type of seeds might be induced during the first 6 and 12 months of storage. However, seeds stored for longer periods of time might also be thought to enter into the primary phases of the ageing process where early alterations occur, including the loss of germination-rate and germination-inhibitor contents in the seed coat, together with an incapacity for the seeds to increase their protein and nucleic acid levels during the germination process.

DNA↗

[Postradiation recovery in Pichia pinus yeasts].

A study was made of the postirradiation recovery of yeast Pichia pinus exposed to gamma-quanta and alpha-particles. No recovery was noted in the haploid cells: the nonreversible component k = 1. In the diploid cells, the nonreversible component k = 0.7 and k = 0.8 after exposure to gamma-quanta and alpha-particles, respectively.

Alpha Particles↗

Germination inhibitors in embryos and endosperms from Pinus pinea seeds.

Inhibitors from Pinus pinea L. seed embryos and endosperms were separated using thin layer chromatography and a Sephadex G-10 column. The inhibitory activity was resolved into several fractions. One of these behaved similarly to abscisic acid in thin layer chromatography and Sephadex G-10 chromatography and in UV absorption and fluorescence spectra. In endosperm another potent inhibitor was detected, which showed a characteristic syndrome similar to that of phosphon D. These germination inhibitors are involved in the regulation of P. pinea seed germination.

Abscisic Acid↗

[Changes in the albumin and globulin fractions in seeds of Pinus pinea during artificial aging].

The alterations on the electrophoretic patterns of albumin and globulin fractions in both embryo and endosperms of Pinus pinea seeds during artificial ageing have been investigated. The results show that the electrophoretic patterns of aged seeds are drastically different from those of seeds with high germination capacity. These alterations were observed even when the quantitative analysis did not show any significant changes.

Aging↗

[Changes in proteins and proteases during artificial aging in seeds of Pinus pinea].

Changes in the germination-rate and the biochemical differences in soluble proteins and proteases in artificially aged Pinus pinea seeds have been investigated. In the aged seeds, soluble proteins decrease during the germination process, parallel to increased levels of proteases activities in both endosperms and embryos. These changes might be involved in the loss of viability.

Peptide Hydrolases↗

On the stability of isocitrate lyase from Pinus pinea.

Isocitrate lyase is a key catalyst of the glyoxylate cycle. A feature of the enzyme from higher plants is the high instability, that causes innumerable problems in working for characterization of the enzyme. The present communication demonstrates that the optimal conditions for the storage of isocitrate lyase from Pinus pinea are: the use of a low temperature (possibly below -20 degrees C), the realization of a high endogenous protein concentration of the enzyme preparations, or, above all when long storage conservation is necessary, the preservation of the enzyme in dried form (acetone precipitation), under vacuum at 4 degrees C. The data reported in this paper seem to exclude, in the above studied conditions, a role for serine proteases in the destabilization of the enzyme. The thiol compounds are not determinant and no effect is obtained by adding exogenous proteins (serum albumin, beta-fructosidase).

Ammonium Sulfate↗

[d-Aminolevulinic acid dehydrase in germinating seeds of Pinus pinea].

The purification grade of d-aminolevulinic acid dehydrase, the specific enzyme activity, the Michaelis constant and the optimum pH are determined in extracts from embryos and endosperms of Pinus pinea seeds germinated at different stages of germination. On the other hand the influence of levulinic acid, sodium cyanide, sodium malonate, sodium arsenite and sodium azide on the activity of d-aminolevulinic acid dehydrase has also been studied.

Aminolevulinic Acid↗

[Autoselection of Pichia pinus yeasts by 3-stage multiflow cultivation].

Three-stage multiflow cultivation of the yeast Pichia pinus, with the flow rates of 0.27, 0.6 and 1.2 hr-1, respectively, in the first, second and third fermenters, resulted in autoselection and the parent strain was substituted by a selective one in the third fermenter. The selective strain differed from the parent one by the size of cells, had a higher specific growth rate, and contained more protein. These properties of the selective strain were preserved in the course of one-stage cultivation.

Ascomycota↗

[Comparative study of the physiological and biochemical characteristics of haploid and diploid Pichia pinus strains in their growth process on a glucose medium].

The economic coefficient, the growth rate, the quantitative content of bound amino acids, and the activities of hexokinase, pyruvate kinase, phosphorylase, malate dehydrogenase and glucose-6-phosphate dehydrogenase were studied comparatively in the haploid and diploid strains of Pichia pinus in the process of their growth in a medium containing glucose. An increase in the content of DNA in the cells was found to be accompanied with an increase of the specific growth rate at the exponential phase of growth and an increase in the activities of hexokinase and glucose-6-phosphate dehydrogenase; however, it had virtually no effect on the economic coefficient, the amino acid composition of protein, and the activities of pyruvate kinase and malate dehydrogenase.

Amino Acids↗