Diurnal regulation of NO3- uptake in soybean plants III. Implication of the Dijkshoorn-Ben Zioni model in relation with the diurnal changes in NO3- assimilation

According to the Dijkshoorn-Ben Zioni model, NO₃^– uptakein the roots is stimulated by NO₃^– assimilation in theshoots, through downward phloem transport of malate synthesizedin response to reduction of NO₂^– to NH³. In this paper,one hypothesis resulting from this model was tested, i.e. thatthe diurnal changes in NO₃^– uptake are due to the lightdependence of NO₃^– reduction in the leaves. This dependencewas studied in detached leaves transferred to deionized wateror supplied via the transpiration stream with similar amountsof ¹⁵NO₃^– in light or darkness. In the dark, the reductionof previously stored NO₃^– or xylem-borne ¹⁵NO₃^–was generally about 40–50% of that measured in the light.Glucose supply to the detached leaves stimulated NO₃^–reduction in the dark, but not enough to increase it up to thesame rate as in the light. Nitrite reduction in detached leaveswas much less affected by darkness, and could be maintainedat a high level by exogenous supply of substrate. Advantagewas taken from this last observation to sustain NO₂^–reductionin attached darkened shoots at the same rate as in the light,by ensuring an appropriate delivery of NO₂^– from the xylem.Although this was assumed to restore the light level of theassociated synthesis of malate, it led to a marked inhibitionof NO₃^– uptake. In addition, the direct supply of malateto the shoots or to the roots failed to prevent the decreaseof NO₃^– uptake in darkness. Thus, our conclusion is thatthe mechanisms evoked in the Dijkshoorn-Ben Zioni model do notplay an important role in the diurnal variations of NO₃^–uptake in soybean plants. Key words: Glycine max, light/dark cycle, malate synthesis, NO₃^– reduction, NO₃^– uptake     

Diurnal regulation of NO-3 uptake in soybean plants IV. Dependence on current photosynthesis and sugar availability to the roots

The short-term dependence of NO^–₃ uptake upon photosynthesisand sugar supply to the roots of soybean plants was investigatedin a series of experiments where CO₂ availability, light intensityor conduction of phloem sap to the roots were severely limited.Removal of CO₂ from the atmosphere or girdling of the stem equallyprevented the stimulation of NO^–₃ uptake when plants weretransferred from darkness to the light. The effect of thesetwo treatments can be reversed by CO₂ re-supply or by additionof 10 mM glucose in the nutrient solution, respectively. Glucosewas also more effective in stimulating NO^–₃ uptake byintact plants in darkness than in light. Collectively, theseobservations are interpreted as evidence that the diurnal changesin NO^–₃ uptake are due to decreased phloem transport ofphotosynthates in darkness. Accordingly, the magnitude of thesechanges was much dependent on starch accumulation in the leavesat the end of the photo-period. Shading the plants lowered thisaccumulation, and resulted in an amplification of the diurnalchanges in NO^–₃ uptake. These results are discussed inconnection with the hypothesis that the carbon-dependent plasticityof the night/day ratio of NO^–₃ uptake is an importantfeature of the co-ordination of the acquisition of N and C bythe plant. Key words: Glycine max, light/dark cycle, NO^–₃ uptake, C and N acquisition     

Reversibility of thiamine deficiency-induced partial necrosis and mitochondrial uncoupling by addition of thiamine to neuroblastoma cell suspensions

Culture of neuroblastoma cells in the presence of low thiamine concentration (6 nM) and of the transport inhibitor amprolium leads to the appearance of signs of necrosis: the chromatin condenses, the oxygen consumption decreases and is uncoupled, the mitochondrial cristae are disorganized, the thiamine diphosphate-dependent dehydrogenase activities are impaired. When 10 µM thiamine are added to these cells, the basal respiration increases, the coupled respiration is restored and mitochondrial morphology is recovered within 1 h. Addition of succinate, which is oxidized via a thiamine diphosphate-independent dehydrogenase, to digitonin-permeabilized cells immediately restores a coupled respiration. Our results suggest that the slowing of the citric acid cycle is the cause of the biochemical lesion induced by severe thiamine deficiency and that part of the mitochondria remain functional. (Mol Cell Biochem 174: 121–124, 1997)     

Temporal Patterns in Sheep Fetal Heart Rate Variability Correlate to Systemic Cytokine Inflammatory Response: A Methodological Exploration of Monitoring Potential Using Complex Signals Bioinformatics

Fetal inflammation is associated with increased risk for postnatal organ injuries. No means of early detection exist. We hypothesized that systemic fetal inflammation leads to distinct alterations of fetal heart rate variability (fHRV). We tested this hypothesis deploying a novel series of approaches from complex signals bioinformatics. In chronically instrumented near-term fetal sheep, we induced an inflammatory response with lipopolysaccharide (LPS) injected intravenously (n = 10) observing it over 54 hours; seven additional fetuses served as controls. Fifty-one fHRV measures were determined continuously every 5 minutes using Continuous Individualized Multi-organ Variability Analysis (CIMVA). CIMVA creates an fHRV measures matrix across five signal-analytical domains, thus describing complementary properties of fHRV. We implemented, validated and tested methodology to obtain a subset of CIMVA fHRV measures that matched best the temporal profile of the inflammatory cytokine IL-6. In the LPS group, IL-6 peaked at 3 hours. For the LPS, but not control group, a sharp increase in standardized difference in variability with respect to baseline levels was observed between 3 h and 6 h abating to baseline levels, thus tracking closely the IL-6 inflammatory profile. We derived fHRV inflammatory index (FII) consisting of 15 fHRV measures reflecting the fetal inflammatory response with prediction accuracy of 90%. Hierarchical clustering validated the selection of 14 out of 15 fHRV measures comprising FII. We developed methodology to identify a distinctive subset of fHRV measures that tracks inflammation over time. The broader potential of this bioinformatics approach is discussed to detect physiological responses encoded in HRV measures.     

Early Stages of Conjugation in Escherichia coli

We initiated these studies to learn more about the initial events during bacterial conjugation and to optimize conditions for their occurrence. We found that cells in donor cultures grown anaerobically prior to mating have (i) a higher mean number of F pili per cell, (ii) longer F pili, (iii) a higher probability of forming specific pairs with F(-) cells, and (iv) a faster rate of initiation of chromosome transfer than cells grown aerobically. The growth medium for the donor culture also influences these same parameters: a rich medium is superior to a completely synthetic medium. Starvation of donor cells in buffered saline or for a required amino acid results in (i) a loss of F pili, (ii) a loss in the ability of donor-specific phages to adsorb, (iii) a loss of ability to form specific pairs with F(-) cells and to yield recombinants, and (iv) an increase in recipient ability. These changes occur as a function of starvation time, and at rates which are dependent on the conditions of prior growth and starvation of the donor culture. Either treatment provides a rapid method for the production of F(-) phenocopies from donor cultures. Resynthesis of F pili by cells within a starved donor culture commences very soon after restoration of normal growth conditions, but full restoration of donor ability, as measured by recombinant yield, occurs at a slower rate. We found, along with other investigators, that F pili are essential for specific pair formation. We also found, however, that the presence of F pili is not sufficient for display of donor ability, nor is the absence of F pili enough for cells to exhibit recipient ability. This suggests, therefore, that one or more components, in addition to F pili, are necessary for the conversion of specific pairs to effective pairs (or for chromosome mobilization, or both) and for preventing donor cells from acting as recipients. On the basis of our results, we suggest optimal conditions for achieving high mating efficiencies.     

Microinjection of glycosaminoglycan-degrading enzymes in the chicken blastoderm

The relationship between the presence of glycosaminoglycans (GAGs) and the morphology of the middle layer or mesoblast was examined by performing transmission electron microscopy of chicken blastoderms microinjected with GAG-degrading enzymes. The controls included microinjections with saline or trypsin, as well as solid-phase assays for proteolytic activity in commercially available GAG-degrading preparations. The results indicate that, in normal as well as in saline-injected blastoderms, middle-layer cells are rounded or cuboidal in shape, and are linked to each other by small intercellular junctions in the primitive-streak region. As they migrate laterally along the basal lamina, they appear as typical mesenchymal cells, being separated by large intercellular spaces and covered by cell processes. The removal of hyaluronate (by the microinjection of hyaluronidases) led to compaction of the middle-layer cells in the area lateral to the primitive streak. These cells lost their mesenchymal aspect and retracted their processes, and intercellular junctions were observed. The presence of proteolytic activity in the enzyme preparations did not interfere with the results. On the basis of the results obtained using this microinjection technique, we were able to confirm at the ultrastructural level that hyaluronate, due to its space-creating properties, promotes the detachment of ingressed primitive-streak cells and preserves the mesenchymal aspect of the middle layer during the lateral migration of single cells along the basal lamina. Whether the presence of hyaluronate is necessary to allow positioning of the mesoblast could not be inferred using our experimental procedure. We present evidence that this molecule, as well as having physicochemical properties, is also involved in the modulation of tissue interactions during gastrulation.     

Isolation of an IS1 flanked kanamycin resistance transposon from R1drd19

Summary We have isolated and identified an IS1-flanked transposon from the plasmid R1drd19. This transposon specifies resistance to kanamycin and is 10.4 kb long. It exhibits a frequency of transposition two orders of magnitude lower than that of the smaller, IS1-flanked transposon Tn9. We have named it Tn2350.     

The initiation of chromosome replication in a dnaAts46 and a dnaA+ strain at various temperatures

Summary The regulation of chromosome replication initiation was studied at various temperatures with an E. coli dnaA46 strain and its dnaA ⁺ parent. We find that, in both strains, the initiation mass varies depending upon growth temperature while the replication time remains constant relative to the cell doubling time. In the permissive temperature range, the initiation mass of the dnaA46 mutant strain is larger by a constant factor than that for a dnaA ⁺ strain. We conclude that, even at temperatures permissive for growth of the dnaA46 strain, the activity of the dnaA46 product is lower than that of the wild-type protein. The dnaA gene product, therefore, plays an important role in regulating initiation.     

Evidence for Anion-Permselective Membrane in Crayfish Muscle Fibers and Its Possible Role in Excitation-Contraction Coupling

Under certain conditions only, isolated crayfish skeletal muscle fibers change in appearance, becoming grainy, darkening, and seemingly losing their striations. These changes result from development of large vesicles on both sides of the Z-line. The longitudinal sarcoplasmic reticulum remains unaffected. The vesicles are due to swelling of a transverse tubular system (TTS) which is presumably homologous with the T-system tubules of other muscle fibers. The vesiculations occur during efflux of water or on reducing external K or Cl, but only when KCl can leave the fiber. They never result from osmotic, ionic, or electrical changes when KCl cannot leave. Inward currents, applied through a KCl-filled intracellular cathode, also cause the vesiculations. These are not produced when the cathode is filled with K-propionate, nor by outward or longitudinal currents. Thus the transverse tubules swell only when Cl leaves the cell. Accordingly, their membrane is largely or exclusively anion-permselective. These findings also indicate that the TTS forms part of a current loop, connecting with the exterior of the fiber probably through radial tubules (RT) possessing membrane of low conductivity. Thus, part of the current flowing inward across the sarcolemma during activity can return to the exterior through the membrane of the TTS. The structure and properties of the latter offer the possibility for an efficient electrical mechanism to initiate excitation-contraction coupling.