Isolation of Plant Gene Space-Related Sequence Elements by High C+G Patch (HCGP) Filtration: Model Study on Rice

For the isolation of gene space representative sequence elements, a new methodology—high C+G patch (HCGP) filtration—has been developed using rice as a model. The method is based on the fragmentation of the genomic DNA by methylation-sensitive HpaII and MspI restriction endonucleases having exclusively G/C base pair-containing recognition sites. These enzymes fragment the genome at high C+G content and hypomethylated regions. Cloning fragments spanning such regions in close vicinity (200–2,000 bp) revealed that about 60% of the clones represented gene space sequences resulting in twofold enrichment of these sequences, which is close to the theoretical maximum in rice. The sequence information of clones used in the present study was deposited in the NCBI database under the accession numbers EI 365676–EI 366364.     

Stabilization of the neutral protease of Bacillus stearothermophilus by removal of a buried water molecule.

Using site-directed mutagenesis, Ala166 in the neutral protease of Bacillus stearothermophilus was changed into Ser. Model building and molecular dynamics simulations of the mutant enzyme indicated that the Ser hydroxyl group fits well in a cavity which contains a water molecule in the wild-type enzyme. The Ala166----Ser mutation was expected to exert a stabilizing effect because of the gain in entropy resulting from the release of water molecule from the folded protein to the solvent. In addition, the hydrogen-bonding network around residue 166 was improved upon the mutation. As a result of this mutation the thermostability of the neutral protease was increased by 1.2 +/- 0.1 degrees C.     

Influence of ethylene on nucleic acid synthesis in etiolated Pisum sativum

Ethylene applied to intact etiolated seedlings of Pisum sativumcv. Alaska inhibits incorporation of ³H-thymidine into DNA insubsequently excised plumular and subapical tissue segmentsbut has no influence on incorporation of ³H-uridine into RNA.The effect on DNA synthesis begins about 2 hr after ethyleneis applied, and intensifies progressively. A similar inhibitionof DNA synthesis occurs when ethylene is applied directly toplumular sections cut from control plants, but not with subapicalsegments under these conditions. Inhibition of DNA synthesisby ethylene is reversed by benzyl adenine in plumular sections.Brief exposure of dark grown seedlings to red light causes asubsequent increase in DNA synthesis in plumular tissue. Thechanges in DNA synthesis in tissues exposed to ethylene, benzyladenine and red light are correlated with the effects of thesetreatments on the mitotic index. (Received March 12, 1973; )     

The reaction mechanism of nitrosothiols with copper(I)

Copper and other transition metal ions and their complexes are catalysts for the decomposition of nitrosothiols. In this way they catalyze the biological functions of nitrosothiols. The kinetics and mechanism of the reaction of two nitrosothiols, S-nitrosothiolactic acid and S-nitrosoglutathione (GSNO), with copper(I) are reported. The kinetics of the reaction of Cu(MeCN) _n ⁺ (n=0–3) with the nitrosothiols were studied. The results indicate that Cu⁺ _aq is the active species in the GSNO system, with k(Cu⁺ _aq+GSNO)=(9.4 ±2.0)×10⁷ dm³ mol⁻¹ s⁻¹. The results also indicate that the Cu(MeCN) _n ⁺ (n=0–3) complexes react with S-nitrosothiolactic acid. Transient species are formed in these processes. The results suggest that these species contain copper(I) and thiol. The results shed light on the catalytic role of copper complexes in the decomposition of S-nitrosothiols. Received 10 April 1999 / Accepted 17 December 1999     

Physiological and nutritional responses of beef steers exposed to stable flies (Diptera: Muscidae).

Physiological and nutritional responses were measured in beef steers exposed to laboratory infestations of stable flies, Stomoxys calcitrans (L.). Calves (n = 6 per treatment) were infested with four levels of stable flies for three periods daily (15 min each period) for 14 d. Steers exposed to 0, 10, 20, or 30 stable flies consumed similar amounts of feed and water; they excreted similar amounts of feces and urine throughout the study. During the experiment, changes in body weight were not influenced by treatment. Heart and respiration rates, rectal temperatures, and circulating cortisol concentrations were similar among groups. Finally, nitrogen consumed, nitrogen lost in feces and urine, and nitrogen retained were unaffected by treatment. We suggest that intermittent exposure to these levels of stable flies, under the conditions of this study, does not evoke physiological and nutritional responses in beef steers.     

Field and laboratory efficacy studies of erythrosin B for Musca domestica (Diptera: Muscidae) and Drosophila robusta (Diptera: Drosophilidae) control

A 1.0% liquid bait formulation of erythrosin B was tested for house fly, Musca domestica L., control in one room of an environmentally controlled caged-layer poultry facility. Reduced fly numbers were recorded between day 14 and day 17 in the room treated with erythrosin B, after 17 d populations increased significantly in treatment and control rooms. Increasing the light intensity to 188 or 386 lm/m squared for 8 h a day or increasing bait stations from one station per 87 m3 to one station per 70 m2 had no effect on developing house fly populations. Annoying levels of a vinegar fly, Drosophila robusta group, developed in the poultry facility during the study. Populations increased to greater than 400 adults/m2 of wall surface by day 15, then declined in the treatment room to less than 1 adult/m2 by day 33. Mortality of house flies that had ingested a 1.0% liquid formulation of erythrosin B was recorded under laboratory light intensities of 51, 532, and 1,030 lm/m2. All three intensities resulted in mortality significantly higher than the control, but mortality among the light intensity treatments did not differ significantly. When house flies were supplied water with erythrosin B, mortality did not differ significantly from that of the control.