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131.
Acute toxicity and degradation of an aqueous suspension of Thiram were studied in Cloeon dipterum. Lethal concentrations for 50% of the larvae (LC 50) after 24 to 96 h of exposure and thresholds were calculated. LC 50 varied from 0.39 to 1.92 mg·l?1 according to the time and the physiological state of the larvae. It was demonstrated that: the sensitivity of the larvae increased with the time during which they were kept in the laboratory; while the toxicity of the aqueous suspension (1 mg·l?1) of Thiram decreased with time, another toxic product was formed. 相似文献
132.
Influence of phospholipase treatments on ligand bindings to a benzodiazepine receptor-GABA receptor-chloride ionophore complex 总被引:2,自引:0,他引:2
Treatment of rat cerebellar membranes with phospholipase A2 (PLA2) or phospholipase C (PLC) increased basal [3H]diazepam binding at 0 degrees C with concomitant disappearance of the stimulatory effect of Cl- ion on the binding. On the other hand, these treatments did not affect the stimulatory effect of GABA, nor the maximum enhancement obtained in the presence of both GABA and Cl- ion. These results suggest that PLA2 or PLC modified the phospholipids responsible for the interaction between the benzodiazepine receptor and the Cl- ionophore. This assumption was supported by the results of thermodynamic experiments which showed that the changes in thermodynamic parameters occurring after the addition of Cl- ion resembled those after PLA2 or PLC treatment. Since the effect of PLA2 was evident at very low concentrations, and a PLC concentration of at least one order of magnitude higher was required to induce a similar effect, the change of phospholipids especially to lysophospholipids seems to be of particular importance. Protein release from the membrane, which also occurs after PLA2 or PLC treatment, did not appear to be responsible for the present phenomenon. 相似文献
133.
The aim of this work was to identify proteins specific for plant cell membranes which could then be used as unique markers. A crude membrane fraction was isolated from corn coleoptiles and separated on non-linear sucrose density gradients. Separation of endoplasmic reticulum (NADH-cytochrome c reductase), mitochondria (cytochrome c oxidase), golgi (inosine diphosphatase), and plasma membranes (N-1-naphthylphthalamic acid-binding) was achieved. The membrane proteins from the gradient fractions were separated using sodium dodecyl sulphate-poly-acrylamide gel electrophoresis and the gels stained with coomassie blue or with concanavalin A/peroxidase to detect glycoproteins. Proteins specific for the various membranes were identified. Five proteins including two glycoproteins were plasma membrane markers. Protoplasts were isolated and iodinated using lactoperoxidase/glucose oxidase covalently attached to beads. Eleven iodinated proteins were found and three of these corresponded to proteins specifically associated with plasma membranes in the density gradients. Two methods for detecting Ca2+-binding proteins following sodium dodecylsulphate polyacrylamide gel electrophoresis were employed. The majority of such proteins were found in the endoplasmatic reticulum and one was specific for plasma membranes. In vitro and in vivo phosphorylation of membrane proteins was examined and the majority of proteins phosphorylated were glycoproteins. Two of the phosphorylated proteins (Mr=110,000 and 20,000) were also iodinated on protoplasts and may be part of the plasma membrane ATPases.Abbreviations ER
endoplasmic reticulum
- IDP
inosine diphosphate
- NPA
N-1-naphthylphthalamic acid 相似文献
134.
We have compared the partial nucleotide and derived amino acid sequences of a phaseolin seed storage protein gene ofPhaseolus vulgaris (1) and a conglycinin storage protein gene ofGlycine max (2). Although these proteins are not antigenically related to one another, the architecture of the genes is similar throughout the sequences compared here. Intervening sequences interrupt the same amino acid positions in both genes. Within the 28% of theG. max gene and the 38% of theP. vulgaris gene represented in this comparison, 73% of the nucleotides in the coding and intervening sequences are identical, excluding the insertions and deletions. The nucleotide mismatches found in the coding sequences are distributed throughout the three codon positions with little bias towards the third codon position. In addition to the single nucleotide differences, six insertions or deletions, ranging from three to twenty-seven nucleotides in length, occur in this portion of the coding region and these are partially responsible for the molecular weight differences of the conglycinin α′-subunit and the phaseolin subunit. 相似文献
135.
The glutamine analog, 6-diazo-5-oxo-l-norleucine (DON), a glycoconjugate inhibitor, was used to probe the relationships between myocardial secretion of extracellular matrix and endothelial differentiation and formation of cushion mesenchyme (primordia of AV values). When DON was given to stage 12 chick embryos maintained in shell-less culture, the myocardial secretion gradient of glucose- and sulfate-labeled matrix was blocked. Concomitantly, the endothelium failed to complete activation but continued to divide and incorporate thymidine. By varying DON concentration, two distinct phases of endothelial differentiation were identified: the first (labile to 0.5 μg) involved hypertrophy, the second (labile to 0.25 μg) acquisition of migratory appendages with resultant mesenchyme formation. Glucosamine + DON (but not inosine, glucose, or glutamine) restored the matrical secretion gradient and to varying degrees both phases of endothelial activation. Endothelia totally suppressed from forming mesenchyme in situ acquired this capacity when explanted into three-dimensional collagen gel culture. The capacity was enhanced by glucosamine given in situ as an inhibitory override, dependent upon serum concentration, inhibited by heat-inactivated serum or by adding DON to the medium, but unaffected by hyaluronate. These results were compared to those obtained by co-culturing endothelium and myocardium and discussed in terms of the hypothesis that cushion mesenchyme formation results from an epithelial interaction mediated by glycoconjugates. 相似文献
136.
The interaction between lipid peroxidation and prostaglandin synthesis in rabbit kidney-medulla slices. 总被引:1,自引:0,他引:1 下载免费PDF全文
Lipid peroxidation induced by ascorbic acid and Fe2+ was inhibited by mepacrine (phospholipase A2 inhibitor) and aspirin (prostaglandin cyclo-oxygenase inhibitor) in rabbit kidney-medulla slices. Moreover, ascorbic acid and Fe2+ potentiated the inhibitory effect on prostaglandin E2 formation by mepacrine, but they had no influence on prostaglandin E2 production decreased by aspirin. Lipid peroxidation induced by ascorbic acid and Fe2+ appears to be affecting the activity of prostaglandin endoperoxide synthase. These results suggest that lipid peroxidation is connected closely with the prostaglandin-generating system, and it has the potential to modulate the turnover of arachidonic acid and prostaglandin synthesis. 相似文献
137.
Roger N. Rosenberg 《In vitro cellular & developmental biology. Plant》1972,8(3):194-204
Summary Newborn BALB/c mouse brain was cultured as disaggregated cells after serial trypsin dissociations. The ontogeny of the cultures
was followed by assays of cell number, deoxyribonucleic acid, and protein content and by the activities of three enzymes considered
to be markers of neuronal differentiation. Aliquots of the freshly dissociated cells were assayed for choline acetylase, acetylcholinesterase,
and glutamic acid decarboxylase activities and compared with intact brain. The percentages of recovery of activities, expressed
as14C product formed per mg of protein per 10 min, at pH 6.8 and 37°C, were 37% for choline acetylase, 54% for acetylcholinesterase,
and 24% for glutamic acid decarboxylase. The remainder of the freshly dissociated cells were placed into culture; enzyme assays
were performed as the cells multiplied and then when the cultures became static. Choline acetylase activity increased as the
cells rapidly divided, and glutamic acid decarboxylase activity increased only after the cultures became confluent. Under
the culture conditions, acetylcholinesterase was not induced, despite active synthesis of acetylcholine.
Neuroblastoma clone N18, C1300 cell line, was grown in cell culture, and the activity of acetylcholinesterase was measured
as the cells multiplied and came to confluency. The specific activity of mouse neuroblastoma acetylcholinesterase increased
25-fold when the rate of cell division was restricted. The rate of cell division could be regulated by adjusting the serum
concentration. By removing fetal calf serum during the growth period, cell division ceased, and acetylcholinesterase activity
was significantly and rapidly induced. Choline-O-acetyltransferase specific activity was measured in rapidly dividing and in static cultures. Its specific activity was highest
in nondividing cultures, compared to cultures containing actively dividing cells (6-fold), and the specific activity of thymidylate
synthetase was increased 2.5-fold in actively dividing cultures, compared to static cultures.
Glioblastoma cells obtained from the rat astrocytoma, clone C6, were grown in culture, and glucose metabolism was measured
in control cultures, and in cultures containing norepinephrine (0.017 mg per ml). Norepinephrine produced a 50% inhibition
in the incorporation ofd-[14C]glucose. Cells incubated for 2 hr in the presence ofd-[14C]glucose, washed and then incubated in control medium or in medium containing norepinephrine, resulted in the release of
greater than 50% of radioactive metabolites in the norepinephrine treated plates. Norepinephrine caused a 50% increase in14CO2 production in glioblastoma cells incubated withd-[1-14C]glucose. Norepinephrine, under similar conditions, did not affect the metabolism of glucose in clone C46, C1300 mouse neuroblastoma
cells.
Portions of this work were supported by a research grant (6-444946-58605) from the American Cancer Society. 相似文献
138.
Kinetics of the activation of rat liver pyruvate kinase by fructose 1,6-disphosphate and methods for characterizing hysteretic transitions 下载免费PDF全文
H. Olin Spivey Wayne Flory Benigno D. Peczon John P. Chandler Roger E. Koeppe 《The Biochemical journal》1974,141(1):119-125
1. Kinetics of fructose 1,6-diphosphate activation of liver pyruvate kinase type I inhibited with MgATP and l-alanine are described as a function of enzyme and fructose 1,6-diphosphate concentrations. These results can be explained by a single pseudo-first-order transition of the enzyme into an active form, independent of the enzyme concentration. This rate constant, k(app.)=0.24s(-1) with 0.02mm-fructose 1,6-diphosphate (t(0.9) approximately 10s where t(0.9) is the time for 90% conversion), is an increasing function of fructose 1,6-diphosphate concentration far beyond that needed to maximally activate enzyme equilibrated with fructose 1,6-diphosphate (about 20mum). 2. The model equations are best analysed with numerical techniques which are described. These techniques are useful in studying similar slow transients frequently observed in stopped-flow studies of enzymes. 3. Shorter transients (t(0.9)=0.5-1.5s) were observed in the kinetic response of the enzyme to the addition of MgATP or phosphoenolpyruvate, but were not further characterized. 相似文献
139.
S-(+)-3,4-Dihydroxybutylphosphonic acid, an isosteric analogue of sn-glycerol 3-phosphate, was synthesized stereospecifically and shown to be an effective substrate for rabbit muscle glycerol 3-phosphate dehydrogenase (sn-glycerol 3-phosphate-NAD(+) oxidoreductase, EC 1.1.1.8). Non-isosteric phosphonate analogues of sn-glycerol 3-phosphate showed neither substrate nor inhibitory activity with the enzyme. 相似文献
140.
The effects of potassium, 5-hydroxytryptamine, adrenocorticotrophin and angiotensin II on the concentration of adenosine 3′:5′-cyclic monophosphate in suspensions of dispersed rat adrenal zona glomerulosa and zona fasciculata cells 总被引:4,自引:1,他引:3 下载免费PDF全文
Janet D. M. Albano Barry L. Brown Roger P. Ekins Sylvia A. S. Tait James F. Tait 《The Biochemical journal》1974,142(2):391-400
Dispersed rat adrenal cells prepared from both the capsule and the decapsulated gland were used to investigate the effects on cyclic AMP accumulation of known stimuli of steroidogenesis [ACTH (adrenocorticotrophin), angiotensin II, K(+) ions and 5-hydroxytryptamine]. Since glomerulosa-cell preparations from capsular strippings are normally contaminated with a proportion of fasciculata cells, cells purified by fractionation on a bovine serum albumin gradient were also used. The results showed that: (1) ACTH and angiotensin II stimulated cyclic AMP accumulation in both fractionated and unfractionated zona fasciculata cells; (2) 5-hydroxytryptamine and an increased extracellular K(+) concentration (from 3.6 to 8.4mm) had no effect on cyclic AMP concentrations in fasciculata cell preparations; (3) the addition of ACTH, angiotensin II, 5-hydroxytryptamine or K(+) to the incubation medium resulted in increased cyclic AMP concentrations in unpurified zona glomerulosa cell preparations; (4) fractionation and hence the virtual elimination of fasciculata contamination, did not affect the response to 5-hydroxytryptamine and increased K(+) concentration. However, the responses to ACTH and angiotensin II were markedly lowered but not abolished. These results strongly suggest a link between cyclic AMP production and steroidogenesis in the zone of the adrenal gland that specifically secretes aldosterone. All four agents used stimulated both steroid output and cyclic AMP accumulation. However, at certain doses of 5-hydroxytryptamine, K(+) and angiotensin II the significant increases in corticosterone output were not accompanied by measurable increases in cyclic AMP accumulation. 相似文献