Activity of cell wall-associated enzymes in ripening olive fruit

Enzymatically active cell wall isolaled from olive (Olea europaea) fruit was employed Hi investigate some hydrolytic enzymes bound to the cell wall and the changes in these during ripening. Seven glycosidases. β-glucosidase (EC 3.2.1.21) α-galactosidase (EC 3.2.1.22). β-galactosidase (EC 3.2.1.23). α-arabinosidase (EC 3.2.1.55), α-mannosidase (EC 3.2.1,24). β-xylosidase (EC 3.2.1.37) and β-N-acetylglucosamidase (EC 3.2.1.30). as well as Cx-cellulase (EC 3.2.1.4) and endo-polygalacturonase (EC 3.2.1.15). were identified in the cell wall preparation, at four stages of ripeness (mature green. changing colour, black and black-ripe). Activities of all these cell wall-associated enzymes fionicallv and covalently linked) were determined either by cell wall incubation with artificial substrate or after extraction from the cell wall with buffers of high salt concentration (Cx-cellulase). and were compared to those of forms solubilized from acetone powders with 500 nM citrate buffer (cytoplasmic and/or apoplastic plus ionically hound to cell wall) In general, the activities of low ionic strength buffer-soluble enzymes were found to be much higher than those of the bound enzymes. The bound enzymes are present in the fruit at the green colour stage, whereas the activities of the soluble enzymes only increased from the changing colour stage onwards. The tenacity of binding of enzymes to the wall was investigated by treating the walls with high salt and measuring residual activity. The nature of the ionic and covalent binding and the changes during ripening were also established for wall-hound glycosidase During ripening there was a marked change in the percentages of covalently- and tonically linked activities of β-glucosidase and β-galaclosidase: al the changing colour stages about 75–80% of the bound active in was present in high ionic strength buffer while al the black-ripe stage it was only 15–20. A possible role for these cell wall degradative enzymes in olive softening is discussed.     

Mixotrophic productivity of the marine diatom Phaeodactylum tricornutum cultured with soluble fractions of rye,wheat and potato

The marine microalga Phaeodactylum tricornutum was cultured semi-continuously with the soluble fractions of wheat, rye and boiled potato flours. Fifteen percent of the culture volume was renewed every 3 d. The cell productivities were 0.9×109 cells/l/d, 1.1×109 cells/l/d and 2.6×109 cells/l/d for wheat, rye and potato respectively. The productivity of the autotrophic control was 1.0×109 cell/l/d. When a soluble fraction of raw potato was added, the productivity was enhanced to 4.1×109 cells/l/d, 2.4 times higher than the autotrophic culture. The high productivity of P. tricornutum with the soluble fractions of Solanum tuberosum suggests its usefulness as a source of nutrients for the production of microalgal biomass.     

Mauthner cell-initiated abrupt increase of the electric organ discharge in the weakly electric fishGymnotus carapo

Stimulation of the spinal cord of the electric fish Gymnotus carapo, evoked an abrupt increase in the discharge rate of the electric organ. At the maximum of this response, the rate increased an average of 26 ± 11.8%. The duration of the response was 4.9 ± 2.12 s; its latency was 10.4 ± 1.1 ms. Activation of the Mauthner axon played a decisive role in this phenomenon as indicated by the following: (1) recordings from the axon cap of the Mauthner cell demonstrated that the response was evoked if the Mauthner axon was antidromically activated and (2) a response that was similar to that produced by spinal cord stimulation, was elicited by intracellular stimulation of either Mauthner cell. Stimulation of the eighth nerve could also increase the discharge rate of the electric organ. The effect was greater if a Mauthner cell action potential was elicited. The findings described in the present report, indicate the existence of a functional connection between the Mauthner cell and the electromotor system in Gymnotus carapo. This connection may function to enhance the electrolocative sampling of the environment during Mauthner-cell mediated behaviors. This is a novel function for the Mauthner cell.Abbreviations EHP extrinsic hyperpolarizing potential - EOD electric organ discharge - M-AIR Mauthner initiated abrupt increase in rate - M-cell Mauthner cell - M-axon Mauthner axon - PM pacemaker nucleus - PM-cell pacemaker cell - PPn prepacemaker nucleus - SPPn sublemniscal prepacemaker nucleus     

Sex hormone changes induced by the parasite lead to feminization of the male host in murine Taenia crassiceps cysticercosis

Female mice are more susceptible to Taenia crassiceps (TC) infection than males. However, after a month parasite load increases massively in both genders reaching thousands of parasites per host. The possibility of hormonal changes in the infected mice was envisaged. Sex hormones levels were assayed after different periods of infection, the parasites present in the peritoneal cavity were collected and gonads, uterus and seminal vesicles were weighed. In male mice, serum estradiol increased to levels 200 times their normal values whilst those of testosterone decreased 90% relative to controls. The weight of seminal vesicles was significantly diminished. Infected female mice also showed a slight increase in estrogen blood levels after 8 weeks of infection and the weight of the uterus was significantly increased relative to controls. Serum estradiol and testosterone were almost undetectable after gonadectomy. Cytokines such as IL-6 are capable of stimulating aromatase activity and we found that splenocytes from infected mice produced amounts of IL-6 higher than control as measured by ELISA. In conclusion T. crassiceps infection triggers a feminization process in the infected hosts. The gonads are required for the parasite to induce higher estrogen synthesis. IL-6 could be involved in the immunoendocrine mechanism used by the parasite to maintain a highly permissive environment for its rapid growth.     

Cellulase production by Rhizobium

Summary The production of cellulase byRhizobium species was studied.Rhizobium trifolii cellulase was induced by a variety of polysaccharides, including celluloses and hemicelluloses. Cellobiose and myo-inositol also allowed enzyme expression but mannitol prevented it at concentrations higher than 0.25%. Both soluble and insoluble plant root substances moderately stimulated cellulase production byRhizobium trifolii.Most substances tested did not induce the production of cellulases by the slow-growing, cowpea type rhizobia strain CIAT 79. Effective inducers were carboxymethylcellulose, gluconate and myo-inositol.Cellulase production was very low under all conditions tested. In most cases the enzyme activity was loosely bound to the capsular material. The enzyme in fast-growers is an 1,4--D-glucan-4-glucanohydrolase (endo-glucanase EC 3.2.1.4) with specificity for high molecular weight polysaccharides.There was no correlation between infectiveness ofRhizobium trifolii strains and cellulase production. One strain, which lacks the nodulation plasmid, produced cellulase at the same rate as its parental infective strain.     

A major substrate of maturation promoting factor identified as elongation factor 1 beta gamma delta in Xenopus laevis

Protein synthesis is believed to be under control of the cell cycle during meiosis and mitosis. Any relationship between substrates for cdc2 kinase and components of the protein synthetic apparatus would therefore be of prime importance. During meiosis of Xenopus laevis oocytes one of the substrates for this kinase is a p47 protein, which is complexed to two other proteins, P36 and P30. Judged from partial amino acid sequence data on P47 and P30, the P30 and P47 proteins were reported to resemble the protein synthetic elongation factors (EF) 1 beta and 1 gamma from Artemia salina (Bellé, R., Derancourt, J., Poulhe, R., Capony, J.P., Ozon, R., and Mulner-Lorillon, O. (1989) FEBS Lett. 255, 101-104). This paper shows that the complex composed of P30, P47, and P36 from Xenopus is identical to the complex of EF-1 beta, EF-1 gamma, and EF-1 delta from Artemia according to two criteria. 1) Both stimulate elongation factor 1 alpha-mediated transfer RNA binding to ribosomes and exchange of guanine nucleotides on elongation factor 1 alpha to a comparable degree. 2) Each of the three subunits of the protein complex P30.P47.P36 from Xenopus shows a structural homology with one of the corresponding subunits of EF-1 beta gamma delta from Artemia. Presumably the phosphorylation of EF-1 gamma, which associates with tubulin at least in vitro, is important in processes following the onset of meiosis which is accompanied by a rise of protein synthesis.     

The influence of ionic composition on the distribution of nematode species in springs of the province of Granada (Spain)

The ionic composition of 38 mineral springs in the province of Granada (Spain), and the distribution of 45 species of nematodes belonging to orders Monhysterida, Araeolaimida, Chromadorida and Enoplida were examined. Water chemistry is used to make two diagrams representing anionic and cationic composition. Diagrams for anionic composition (given the greater variance seen in the springs considered) are used to illustrate the distribution of individual species. The results obtained from species distribution and the correlation between species made it possible to group species which could be associated with springs where each of the anions considered predominated. A greater number of species groups was found to inhabit springs in which chloride concentrations was less than 50% of the total concentration of anions.     

Structure-activity relationships for unsaturated dialdehydes. 6. The mutagenic activity of 11 compounds in the V79/HGPRT assay.

The mutagenic activity of 11 sesquiterpenoid unsaturated dialdehydes in the V79/HGPRT assay has been determined, and is compared with previously published data on the mutagenicity of the same compounds towards Ames Salmonella strains. One compound, isovelleral, is a potent mutagen in both assays, while six compounds, which are positive in the Ames Salmonella/microsome assay, show no significant activity in this study. One compound, acetylmerulidial, is negative in the Ames Salmonella/microsome assay but significantly although weakly mutagenic in the V79/HGPRT assay. The remaining three compounds are inactive in both assays. The study is part of a general investigation of quantitative structure-activity relationships for unsaturated dialdehydes, a class of natural occurring compounds known for their potent and numerous biological activities.     

Evaluation in Drosophila melanogaster of the mutagenic potential of furfural in the mei-9a test for chromosome loss in germ-line cells and the wing spot test for mutational activity in somatic cells.

The mutagenic potential of furfural was evaluated by means of the chromosome loss test in germ cells and the wing spot test in somatic cells of Drosophila melanogaster. The chromosome loss test was carried out employing repair-proficient as well as repair-deficient females. Males carried the compound Y chromosome, BSYy+. Two routes of administration were used: injection and feeding of adult males. Genetic damage was demonstrable after matings of treated males with females carrying the excision repair-deficient mutant mei-9a. The somatic mutation and recombination test was carried out treating 72-h transheterozygous mwh+/+flr3 larvae. Acute treatment of larvae was chosen as the method of exposure. Evidence indicates that furfural induces somatic damage as measured in the wing spot test.