In the present study, oligonucleotides were adsorbed onto the surface of cationic liposomes and nanoparticles at different ratios. As a result, the surface charges of the colloidal carriers were decreased with increasing oligonucleotide concentration. At a certain oligonucleotide concentration, complete charge neutralization led to the aggregation of the carrier systems. Further increasing oligonucleotide concentrations reversed the surface charge of liposomes and nanoparticles to a negative one. Ultrasonication was investigated as a possible means for the homogenization of the formed aggregates. However, the use of ultrasonication led to a time-dependent damage of oligonucleotides adsorbed onto AH-Chol liposomes and MMAEMC-nanoparticles, as well as of unbound oligonucleotides. Nearly 60% of the oligonucleotides adsorbed to MMAEMC-nanoparticles and 65% of ODNs adsorbed to the liposomes were degraded by the effect of cavitation produced by ultrasonication within 10 min. In contrast, the oligonucleotides were protected from degradation when DEAE-stabilized PHCA-nanoparticles were employed as ODN carriers. More than 80% of the oligonucleotides entangled in the surface matrix of these nanoparticles remained intact.

Cyanoacrylates↗

PubMedDate not supplied

Nitrogen fixing activity in Rhizobium japonicum separated from plant cell cultures.

Induced by soy bean tissue cultures in socalled "tissue chambers", Rhizobium japonicum str. 61-A-96 developed nitrogenase activity separated from the plant cells. The activity proceded for 48 h with a rate of 1 X 10(-8) nmol C2H4h-1 cell-1, which is about 6% of the activity measured for bacteroids from Rhizobium japonicum in nodules of Glycine max.

Acetylene↗

PubMedDate not supplied

Differentiation of Rhizobium japonicum, III. Inhibition of nitrogenase derepression by chloramphenicol and rifampicin concentrations, not inhibiting growth.

Development of nitrogenase (40--140 nmol C2H4.mg protein-1.h-1) in Rhizobium japonicum 61-A-101 after transfer to special culture conditions (medium 20 P, 2% O2, 10% CO2, 88% N2 in the gas phase) is inhibited by chloramphenicol (6 X 10(-4)--10(-3) M) and by rifampicin (10(-5) M). These concentrations do not inhibit the slow growth of the cells under these conditions with a doubling time of the cell protein and living cell number of 3--5 d. Nitrogenase activity of previously derepressed cells is not inhibited by chloramphenicol. Growth of the cells under air in yeast extract-mannitol-glycerol medium (8 h doubling time) is affected significantly more by chloramphenicol (2.5 X 10(-4) M) than growth under nitrogenase derepressed culture conditions.

Chloramphenicol↗

PubMedDate not supplied

Differentiation of Rhizobium japonicum, I. enzymatic comparison of nitrogenase repressed and derepressed free living cells and of bacteroids.

Derepressed free living cells of Rhizobium japonicum strain 61-A-101 with leucine as single nitrogen source develop a maximum nitrogenase activity of 180 nmol C2H4.mg protein -1.H-1 in liquid culture under 2% 2% O2 in the gas phase. Only 10% of this activity is found with no oxygen in the gas phase during a 90 min incubation period. The maximum activity under 2% oxygen in the gas phase is unaffected by addition of 1-100 mM NH+4 and by addition of low concentrations of glutamine (0.36-1.44 mM). Specific activities of alanine dehydrogenase (E.C. 1.4.1.1.) asparatate aminotransferase (E.C. 2.6.1.1.) and, with much lower activities, of GOGAT (E.C. 1.4.1.13) in nitrogenase active free living cells are more similar to bacteroids than to nitrogenase repressed free living cells from liquid culture. The activities in nitrogenase repressed cells were about 50% lower. Glutamine synthetase (E.C. 6.3.1.2.) activity in bacteroids and in nitrogenase active cells were also similar, but only about 25-30% of that found in nitrogenase repressed Rhizobium japonicum cells.

Bacterial Proteins↗

PubMedDate not supplied

Screening for mutants of Rhizobium japonicum with defects in nitrogen fixing ability.

Mutants of Rhizobium japonicum with reduced ex planta nitrogenase activity could be isolated with high frequency by direct screening of ultra-violet mutagenized bacteria growing as spots on the surface of an appropriate agar medium permitting derepression of nitrogenase synthesis. Small glass chambers fitted with a serum cap were pushed into the agar around each spot of growth, forming a small enclosed gas space which was made to 10% acetylene, permitting assessment of nitrogenase activity by the acetylene reduction test. Four mutants were isolated from a total of 305 screened spots. Two mutants had almost no ex planta activity, one of which had no symbiotic activity despite normal nodulation (ineffective), the other had only somewhat reduced activity symbiotically. Two other mutants with less than half wild-type activity ex planta were normal in symbiosis.

Mutation↗

PubMedDate not supplied

Biocontrol strain Pseudomonas sp. W34: specific detection and quantification in the rhizosphere of Cucumis sativus with a DNA probe and genotypic characterization by DNA fingerprinting.

A DNA probe specific for biocontrol strains of Pseudomonas was produced by screening randomly amplified polymorphic DNA (RAPD) PCR fragments. Specificity of the probe was assessed by dot blot and colony hybridization. It was used to specifically determine the population of these strains on roots of Cucumis sativus cv. Delikatess. Two polymorphic RAPD fragments of 750 bp, and 550 bp showed identical specificity. The biocontrol strain Pseudomonas sp. W34 was shown to be competitive in the rhizosphere of cucumber and to maintain a stable population for at least 10 days when inoculated on the seed. The phylogenetic relationships between the biocontrol and reference strains were analyzed at the strain level by means of RAPD and repetitive sequence-based PCR genomic fingerprinting (rep-PCR), and at higher taxonomic levels by means of amplified 16S ribosomal DNA restriction analysis ARDRA. It was shown that the antagonistic strains are closely related, forming a separate cluster from other non-antagonistic and reference Pseudomonas strains, their taxonomic placement remaining uncertain.

Cucumis sativus↗

PubMedDate not supplied

Gene expression of Medicago sativa inoculated with Sinorhizobium meliloti as modulated by the xenobiotics cadmium and fluoranthene.

Alfalfa plants (Medicago sativa cv. Europe) inoculated with Sinorhizobium meliloti 2011 (formerly Rhizobium meliloti, de Lajudie et al., 1994) were cultivated for 14 days under standardized growth conditions in mineral medium with addition of the heavy metal cadmium or the polycyclic aromatic hydrocarbon fluoranthene. These xenobiotics significantly reduced the numbers of root nodules before any visible damage to the plant could be detected. EC10, EC50, and EC90 (effective concentrations reducing nodulation, shoot and root fresh weight by 10, 50, or 90% compared to the control without pollutant) were calculated. EC50 for cadmium ranged from 5.8 microM (nodulation) to more than 20 microM (root fresh weight). Testing fluoranthene resulted in an EC50 of 2.5 microg cm(-2) for nodulation, and EC50 values of more than 35 microg cm(-2) for shoot and root biomass production, indicating that the effect parameter nodulation is 10-fold more sensitive than shoot and root fresh weight. With mRNA differential display techniques the effects of both xenobiotics on gene expression in alfalfa root systems were studied. 37 differentially displayed transcripts were detected. Two of them, called DDMs1 and DDMs2, were confirmed by northern hybridization to be down-regulated in the presence of the xenobiotics. The expression of transcript DDMs1 was enhanced in alfalfa control plants inoculated with rhizobia, the transcript level was increased 2.5-3-fold compared to non-inoculated plants. This positive effect of nodulation was suppressed, partly by 35 microg cm(-2) fluoranthene and totally by 20 microM cadmium. The decrease in DDMs1 transcription was highly affected by the cadmium concentration with an EC50 of 5.9 microM. Compared to nodulation, almost identical EC10, EC50, and EC90 values were found for DDMs1 expression. Sequence analysis of DDMs1 revealed a significant overall homology (50% identity) to a hypothetical protein from Arabidopsis thaliana with high similarity to a copper transporting ATPase. High levels of transcript DDMs2 were observed in control plants with a 50% decrease in the xenobiotic-treated plants. DDMs2 gave a strong homology (82% identity) to the cytoplasmatic 60S ribosomal protein L18 from Arabidopsis thaliana.

Amino Acid Sequence↗

PubMedDate not supplied

The struggle for health: from local to global level.

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Community Health Services↗

PubMedDate not supplied

Health care and human dignity.

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Attitude to Health↗