The rep region of pR plasmid regulates the expression of SOS system

Summary By using an artificial hybrid between phage and the pR plasmid, we have shown that the rep region of the pR plasmid encodes a function which regulates the expression of the muc genes (plasmid genes that are under the negative control of lexA and responsible for an increased rate of spontaneous mutagenesis and resistance to UV and chemicals). Expression of the muc genes was monitored by a fusion between the muc promoter and the lacZ structural gene. When E. coli cells containing such a fusion are infected by the hybrid pR phasmid, -galactosidase activity is enhanced, indicating that pR encodes an antagonist of lexA. By deletion mapping we have located the gene encoding the antagonist of lexA (bat) in the rep region of the plasmid. The bat gene product can also antagonize the cI repressor as shown by the observation that pR phasmids are virulent on a homoimmune lysogen. We have exploited this latter property to carry out genetic and functional analysis of the bat region. This region is organized as a classical operon where the expression of the bat structural gene is negatively regulated by a repressor gene that encodes a proteic product.     

Possible change in noradrenergic receptor sensitivity following methylphenidate treatment: Growth hormone and MHPG response to clonidine challenge in children with attention deficit disorder and hyperactivity

Growth hormone (GH) and 3-methoxy-4-hydroxyphenelethylene glycol (MHPG) response was measured hourly for 4 hours in 8 children with Attention Deficit Disorder with Hyperactivity (ADD+H) following an acute single-dose of clonidine. The clonidine challenge was repeated before, during, and one day after 12 weeks of treatment with methylphenidate (MPH). Before MPH treatment, the plasma growth hormone (GH) rose to 31.3 ± 4.6 (Mean ± SE) ng/ml; during MPH treatment, the GH peak was only 14.8 ± 3.2 ng/ml; one day after discontinuation of MPH, GH rose to only 20.8 ± 3.9 ng/ml. MHPG release was inhibited by clonidine in all treatment conditions but tended to be more decreased during MPH treatment. Some children with ADD+H may have hypersensitivity of the post-synaptic alpha-1 noradrenergic receptor which is diminished by MPH treatment. The extent to which these effects are pharmacological or represent a change in receptor sensitivity requires further study.     

Studies on the effects of certain salts on germination,on growth of root,and on metabolism

Summary The effects of different concentrations of KCl, K₂SO₄, MgCl₂ and MgSO₄ on the growth in length of the first seminal root of wheat, and on the change in fresh and oven-dry weight of the seedling and its component parts have been studied. The effect of mannitol was also investigated for comparison and to study the osmotic action. The effect of salts on root growth was dependent on salt species; all effects were specific to ions and not due to osmotic activity of solution. The growth of wheat roots was suppressed by concentrations of salts much lower than those required to suppress germination. All solutions of KCl from 0.1 to 50 me/l checked the growth of the root; the retardation increased with increase of concentration. In K₂SO₄ there was a slight activation of root growth for one day in 0.1 and 0.5 me/l; then the growth was suppressed after that. In all other concentrations from 1 to 50 me/l the growth was retarded. In MgCl₂ or MgSO₄ there was some activation of root elongation in 0.05, 0.1 and 0.5 me/l; but higher concentrations retarded root growth.     

Stabilization of purified potato virus X by dextran T-10 during lyophilization

Purification and preservation of potato virus X from leaf sap of tobacco plants before lyophilization was carried out by two methods: 1) precipitation by polyethylene glycol and ultracentifugation, and 2) precipitation by ammonium sulphate, chromatography on Sephadex G-50 and ultracentrifugation. The first method is preferable to the second because the final preparation contains more virus antigen. Both preparations were strongly infectious and maintained antigenic properties after lyophilization. To achieve a more gentle course of lyophilization of virus preparations, addition of urotropine and dextran T-10 to the virus suspension, purified by the precipitation by polyethylene glycol-6000, was examined. Addition of urotropine was proved unsatisfactory, because only antigenic properties were maintained after lyophilization while the infectivity disappeared. But we can recommend addition of dextran T-10 up to a concentration of 6% to the preparation of virus antigen before lyophilization. The course of lyophilization is much rapider, the lyophilized product can be very easily dissolved in water and is not hygroscopic. The product is strongly infectious and gives the serological precipitation reaction in a dilution four times that of X virus antigen lyophilized without addition of dextran T-10.     

Salinity and hormone interactions in affecting growth,transpiration and ionic relations ofPhaseolus vulgaris

Addition of either abscisic acid (ABA) or kinetin at 10⁻⁶ M to salinized media (20–120mM NaCl) induced remarkable effects on growth ofPhaseolus vulgaris plants. Whereas ABA inhibited the plant growth and the rate of transpiration, kinetin induced stimulation of both parameters. Moreover, ABA increased proline and phosphorus concentrations in the salinized plants whilst kinetin decreased them. ABA induced stimulation of the transport of K, Ca and Cl from root to shoot, accumulation of K, Na and Cl in root cells and inhibits the transport of Na and accumulation of Ca. Kinetin appeared to inhibit the transport and accumulation of Na and Cl, transport of K, and stimulates the accumulation of K and Ca as well as the transport of Ca. The highest influence of both ABA and kinetin was mostly observed when these hormones were used in combination with the highest concentration of NaCl (120 mM) in the medium.     

Interrelationships of Leucine and Glutamate Metabolism in Cultured Astrocytes

Abstract: The aim was to study the extent to which leu-cine furnishes α-NH₂ groups for glutamate synthesis via branched-chain amino acid aminotransferase. The transfer of N from leucine to glutamate was determined by incubating astrocytes in a medium containing [¹⁵N]leucine and 15 unlabeled amino acids; isotopic abundance was measured with gas chromatography-mass spectrometry. The ratio of labeling in both [¹⁵N]glutamate/[¹⁵N]leucine and [2-¹⁵N]glutamine/[¹⁵N]leucine suggested that at least one-fifth of all glutamate N had been derived from leucine nitrogen. At the same time, enrichment in [¹⁵N]leucine declined, reflecting dilution of the ¹⁶N label by the unlabeled amino acids that were in the medium. Isotopic abundance in [¹⁶N]-isoleucine increased very quickly, suggesting the rapidity of transamination between these amino acids. The appearance of ¹⁵N in valine was more gradual. Measurement of branched-chain amino acid transaminase showed that the reaction from leucine to glutamate was approximately six times more active than from glutamate to leucine (8.72 vs. 1.46 nmol/min/mg of protein). However, when the medium was supplemented with α-ketoisocaproate (1 mM), the ketoacid of leucine, the reaction readily ran in the “reverse” direction and intraastrocytic [glutamate] was reduced by ～50% in only 5 min. Extracellular concentrations of α-ketoisocaproate as low as 0.05 mM significantly lowered intracellular [glutamate]. The relative efficiency of branched-chain amino acid transamination was studied by incubating astrocytes with 15 unlabeled amino acids (0.1 mM each) and [¹⁵N]glutamate. After 45 min, the most highly labeled amino acid was [¹⁵N]alanine, which was closely followed by [¹⁵N]leucine and [¹⁵N]isoleucine. Relatively little ¹⁵N was detected in any other amino acids, except for [¹⁵N]serine. The transamination of leucine was ～17 times greater than the rate of [1-¹⁴C]leucine oxidation. These data indicate that leucine is a major source of glutamate nitrogen. Conversely, reamination of a-ketoisocaproate, the ketoacid of leucine, affords a mechanism for the temporary “buffering” of intracellular glutamate.     

Effects of salinity on growth and metabolism ofPhaseolus vulgaris

Increasing salinity induced a marked reduction in the plant growth, thoughPhaseolus seedlings tolerated salinity up to 120 mM NaCI. A great reduction in sugar and protein contents occurred with increasing salinity, whereas soluble nitrogen compounds and the relative contents of the photosynthetic pigments were increased in the treated plants. Increasing Ca concentration in the salinized medium appeared to improve the plant growth and to increase the contents of saccharides and proteins in the NaCl-treated plants. This suggests that Ca could be added to salinized media to overcome the deleterious effects of salinity on the growth and productivity of leguminous crop plants.     

Identification of a locus involved in meningococcal lipopolysaccharide biosynthesis by deletion mutagenesis

A novel method for insertion/deletion mutagenesis in meningococci was devised. This consisted of ligating a digest of total chromosomal DNA to a 1.1 kb restriction fragment containing an erythromycin-resistance marker ( ermC ), and subsequent transformation of the ligation mixture into the homologous meningococcal strain H44/76. Southern blotting of a number of the resulting erythromycin-resistant transformants demonstrated that all carried the ermC gene inserted at different positions in the chromosome. Mutants with a specific phenotype were identified by screening with the anti-lipopolysaccharide (LPS) monoclonal antibody MN4A8B2, which is specific for immunotype L3. In this way, two independent L3-negative mutant strains were isolated. In transformation experiments with chromosomal DNA from these mutants, erythromycin-resistance and lack of MN4A8B2 reactivity were always linked, showing that the insertion/deletion was in a locus involved in LPS biosynthesis. On SDS–PAGE, the mutant LPS displayed an electrophoretic mobility intermediate between that produced by the previously isolated galE and rfaF mutant strains. Chemical analysis of the mutant LPS revealed that the structure was probably lipid A–(KDO)₂–(Hep)₂. Chromosomal DNA flanking the ermC insertion in these two mutant strains was cloned, and used as probe for the isolation of the corresponding region of the wild-type strain. From hybridization and polymerase chain reaction (PCR) analysis, it could be concluded that both mutations map to the same locus. The affected gene probably encodes the glycosyltransferase necessary for adding N -acetylglucosamine to heptose.     

Plastid development in germinating wheat (Triticum aestivum) is enhanced by gibberellic acid and delayed by gabaculine

Etioplast development and protochlorophyllide (Pchlide) accumulation was studied in wheat seedlings ( Triticum aestivum L. cv. Walde, Weibull) grown in darkness on gibberellic acid (GA₃), gabaculine (3-amino-2,3-dihydrobenzoic acid), or on a combination of the two. The results were compared with the features of seedlings grown on water only. GA₃ enhanced shoot growth and promoted etioplast development. A correlation was observed between the appearance of prolamellar bodies (PLBs) and of phototransformable Pchlide. Gabaculine, a known tetrapyrrole biosynthesis inhibitor, delayed growth, slowed down the rate of PLB formation and caused structural alterations of the etioplasts up to 48 h of germination. Gabaculine also delayed the formation of phototransformable Pchlide as well as overall Pchlide biosynthesis, as determined by low-temperature fluorescence emission in vivo. The spectral blue-shift of newly formed chlorophyllide (Chlide) was delayed in irradiated dark-grown gabaculine-grown seedlings, indicating an inhibited dissociation of Chlide and NADPH-Pchlide oxidoreductase (Pchlide reductase: EC 1.3.1.33). Thus there is a close correlation between accumulation of Pchlide and etioplast development, also under conditions when development is enhanced or delayed.