Dehiscence of Coelomomyces psorophorae sporangia from Aedes taeniorhynchus: Induction by amines and amino acids

The effect of various chemicals on induction of dehiscence of nonsterile Coelomomyces psorophorae sporangia from Aedes taeniorhynchus larvae was examined. Tests were made with various salts, reducing agents, chelating agents, buffers, alcohols, carbohydrates, fatty acids and derivatives, amino acids and derivatives, peptides, amines, purines and pyrimidines, antibiotics, and plant hormones. The most active compound was Tris [tris(hydroxymethyl)aminomethane]. Tris was most effective at pH 8–9, and a concentration of 1–20 mM. Structure-activity studies indicated that active compounds had a basic requirement for NH₂ and either COOH or CH₂OH attached to the alpha carbon, but only certain amino acids and amines were highly active. Preparations virtually free of host debris were not responsive to Tris, but addition of bacteria-free homogenates of A. taeniorhynchus larvae restored responsiveness. Dehiscence of C. psorophorae sporangia from Psorophora howardii was also enhanced by exposure to Tris.     

Microsomal palmitoyl-coenzyme A synthetase from rat liver. Partial and exchange reactions

The partial and exchange reactions of long-chain fatty acid activation were determined by using purified microsomal long-chain fatty acyl-CoA synthetase (EC 6.2.1.3). No significant ATP formation from palmitoyl-AMP and PP(i), nor palmitoyl-AMP-dependent CoA disappearance could be demonstrated. Similarly, no palmitate-dependent [(32)P(2)]PP(i)-ATP exchange was catalysed by the pure enzyme. The above partial and exchange reactions were, however, catalysed by the parent microsomal fraction at a rate similar to that of the overall reaction. The implications of these results are discussed.     

Ascorbic acid-2-sulfate sulfhohydrolase activity of human arylsulfatase A.

Pure human arylsulfatase A (EC 3.1.6.1) was found to hydrolyze ascorbic acid 2-sulfate to ascorbic acid and inorganic sulfate at rates from 200 to 2000 mumol/mg per h depending on the method of assay. This rate was lower than that observed with the synthetic substrate 4-nitrocatechol sulfate, but higher than that seen with the physiological substrate cerebroside sulfate. Extracts of cultured fibroblasts from normal subjects were also shown to hydrolyze ascorbic acid 2-sulfate; extracts of fibroblasts from patients with metachromatic leukodystrophy, known to be deficient in arylsulfatase A, did not. Similarly, hydrolysis of ascorbic acid 2-sulfate was not observed when a partially purified preparation of human arylsulfatase B was tested under a variety of conditions. Thus, in the human, arylsulfatase A appears to be the major, if not the only, ascorbic acid-2-sulfate sulfohydrolase.     

Use of polymyxin B, levallorphan, and tetracaine to isolate novel envelope mutants of Escherichia coli.

Mutants of Escherichia coli were isolated by their resistance to the bacteriocidal effects of the membrane-active drugs polymyxin B, levallorphan, and tetracaine. The mutants were examined for additional changes in cellular physiology evoked by the lesions; many polymyxin-resistant strains had a concomitant increased sensitivity to anionic detergents, and several strains of each type had concomitant alterations in generation time and morphology. Mutants of each class (polymyxin resistant, tetracaine resistant, and levallorphan resistant) were transduced into recipient strains. The levallorphan resistance site (lev) was located at approximately 9 min on the E. coli chromosome. Polymyxin (pmx) and tetracaine (tec) resistance loci were also transduced. The lev and tec strains had a slight prolongation of generation time, in contrast with their isogenic wild-type strains. The tec transductant produced long filaments in the absence of tetracaine and had an altered colonial morphology, it reverted at high frequency, with the morphological abnormalities reverting along with the tetracaine resistance. The pmx transductant had an increased sensitivity to levallorphan and to anionic detergents. In contrast, both lev and tec mutants were more resistant to acriflavine than was the wild type or the pmx transductant. The pmx, lev, and tec loci differed in sensitivity to mitomycin C; the lev strain was more resistant, the tec strain was more sensitive, and the pmx strain was much more sensitive than the wild type. There was no difference in sensitivity to several other dyes and detergents, colicins, or T bacteriophage between the transductant and isogenic wild-type strains. Thus, lev, tec, and pmx loci confer more subtle alterations in the permeability barrier than do lipopolysaccharide-deficient mutants previously studied.     

A method for the isolation of cross-linked nucleosides from DNA: application to cross-links induced by nitrous acid.

A procedure is reported for the isolation of cross-linked nucleosides from nitrous acid-treated calf thymus DNA. Cross-linked DNA was hydrolyzed enzymatically with deoxyribonuclease I and snake venom phosphodiesterase and fractionated on a DEAE-Sephadex column. After desalting, the fractions were characterized by ultraviolet spectroscopy, anion exchange high pressure liquid chromatography, gel filtration, and two dimensional thin layer chromatography. A cross-linked dinucleotide, and a series of oligonucleotides were isolated. The oligomers, which had resisted digestion by the above enzyme system, were digested to the nucleoside level by a spleen phosphodiesterase-alkaline phosphatase combination. A second cross-linked product was isolated from this mixture. The cross-linked nucleosides were less than 0.17% of the total nucleotides of the DNA. The methods developed here are recommended for the isolation of products from DNA treated with other cross-linking agents.     

Effect of retinoic acid on chondrocyte glycosaminoglycan biosynthesis.

The effect of retinoic acid on glycosaminoglycan biosynthesis was investigated in rat costal cartilage chondrocytes in vitro. At levels of 10^?9 to 10^?8m retinoic acid, ³⁵SO₄ uptake into glycosaminoglycans was reduced 50%. At these low levels of retinoic acid there was no evidence of lysosomal enzyme release. The results are explained best in terms of modification of glycosaminoglycan synthesis, rather than accelerated degradation. Retinoic acid selectively modified the incorporation of ³⁵SO₄ or [¹⁴C]glucosamine into individual glycosaminoglycans fractions under the conditions studied. The relative incorporation of radiolabeled precursor into heparan sulfate (and/or) heparin increased three- to fourfold. The relative incorporation of radiolabeled precursor remained constant for chondroitin 6-sulfate, whereas incorporation into chondroitin 4-sulfate and chondroitin (and/or) hyaluronic acid decreased. Under the conditions studied, retinoic acid did not appear to be cytotoxic and did exhibit selective control over glycosaminoglycan biosynthesis. It is suggested that the decreased incorporation of ³⁵SO₄ into glycosaminoglycans at hypervitaminosis A levels of retinol may be accounted for by the presence of low levels of retinoic acid, a naturally occurring metabolite.     

The effects of lactose on human erythrocytes

Human erythrocytes suspended in isotonic lactose solution lost potassium and continued to lose potassium even when resuspended in isotonic sodium chloride. The same phenomenon was observed when the cells were suspended in an isotonic solution of the sodium salt of glutamate, a nonpenetrating anion. The presence of 5 mEq per liter of sodium chloride in the lactose or sodium glutamate suspensions greatly reduced the initial potassium loss and the potassium loss when the cells were resuspended in sodium chloride solution. Salts of nonpenetrating anions were less or not effective in blocking lactose damage. The results indicate that absence of penetrating anions in the suspending media is the initiating condition of lactose damage. Chloride and consequently potassium are lost from the erythrocyte. Changes in cellular ionic pattern and/or changes in the cell membrane result in a nontransient damage manifested by continued potassium loss by lactose-treated cells resuspended in isotonic NaCl.     

The effect of the plant cell inhibitor propachlor (alpha-chloro-N-isopropyl-acetanilide) on the cell cycle of L1210 cells as evaluated by flow cytometry

Previously it was reported that the herbicide propachlor (alpha-chloro-N-isopropyl-acetanilide) has a strong inhibitory effect on the proliferation of L1210 mouse leukemia cells. It is now demonstrated that propachlor treatment causes L1210 cells to accumulate in the G1 phase as determined by flow cytometric analysis. This effect of propachlor is dose-dependent with more than 90% of G1 cells accumulating at 10 microM. Kinetic experiments demonstrated that the accumulation of cells in G1 starts in about 10 hours, and increased for up to about 44 hours of incubation with 10 microM propachlor. Treated cells can be revised to a normal DNA distribution by removing propachlor.