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1.
dl-[1,6-14C]Lipoic acid was synthesized and administered to rats or incubated in vitro with rat liver systems. The urinary excretion of radioactivity after labeled lipoate was administered intraperitoneally at a level of 0.5 mg/100 g body weight was maximal at 3–6 hr, with 60% of the injected radioactivity recovered within 24 hr. Respiratory 14CO2 from the same animals is maximal at 3 hr, after which it falls off markedly. Approximately 30% of the injected radioactivity was recovered as 14CO2 within 24 hr. The excretion of radioactivity after lipoate was administered by stomach tube was similar to that after intraperitoneal injection. Localization of radioactivity in the body was greatest in liver, intestinal contents, and muscle in all cases. Ionexchange and paper chromatographies of 24-hr pooled urine revealed several watersoluble radioactive metabolites. Incubation of [14C]lipoate with homogenates or mitochondrial preparations in vitro resulted in the production of 14CO2, which was decreased by incubation with unlabeled fatty acids and unaffected by the addition of carnitine or (+)-decanoylcarnitine. The rat, like certain bacteria, metabolizes lipoate via β-oxidation of the valeric acid side chain and by other metabolic reactions on the dithiolane ring, which render the molecule more water soluble. 相似文献
2.
Yukio Asakawa Kinjiro Tamari Keiko Inoue Jun Kaji 《Bioscience, biotechnology, and biochemistry》2013,77(4):713-717
The localization of tritium-radioactivity in dwarf kidney bean plants (Phaseolus vulgaris) of 3H-gibberellm A3(3H-GA3) applied in a large quantity was investigated in advance of the study on GA3 metabolism in this plant. Immediately after the application of 3H-GA3, the radioactivity was distributed uniformly in the top of this plant; no further transportation of the radioactivity into the growing apical region from mature leaves and stems was the observed as the growth stage proceeded. An investigation on the intracellular localization of the radioactivity demonstrated that most part of the radioactivity was found in the cellular soluble fraction, while no radioactivity was detected in such subcellular particles as nuclei, mitochondria and microsomes. Examinations of the occurrence of GA3 bound with such macromolecules as RNA and protein gave negative results. 相似文献
3.
Evelyne O. Delorme 《Plant Growth Regulation》1987,5(2):125-140
Following application of 3H-Gibberellin A20 (GA20) to roots of G2 pea seedlings and homogenization of the roots, about 3% of the radioactivity in the tissue could be precipitated from a 30,000 × g supernatant with trichloroacetic acid (TCA) (soluble fraction) while about 5% of the radioactivity pelleted at 30,000 × g (particulate fraction). The radioactivity in the particulate fraction was soluble in sodium dodecyl sulfate (SDS), but was not dialyzable and was insoluble in ethanol. Electrophoresis of the soluble fraction gave only one band of radioactivity, while that of the particulate fraction gave multiple bands. Acid hydrolysis of the soluble fraction released radioactivity that ran coincident with acid-treated GA20 on silicic-acid column chromatography. The particulate fraction gave numerous radioactive peaks following acid hydrolysis, two of which were coincident with GA20 and GA29 (hydroxylation product of GA20) on silicic acid chromatography. Treatment of the particulate and soluble fractions with RNase, DNase, and proteases showed a significant solubilization of radioactivity only with the proteases, suggesting that the GA is bound to a proteinaceous macromolecule. Complete proteolytic hydrolyis followed by thin layer chromatography showed 65% of the radioactivity from the soluble fraction running separately from free GAs or the individual amino acids; the particulate fraction gave mainly (60%) free GAs on enzymatic hydrolysis and much smaller amounts (17%) in a position separate from that of the GAs or amino acids. Binding of 3H-GA to protease-sensitive material was obtained with biologically active 3H-GA20 and 3H-GA1. 相似文献
4.
Morten Sandvik Tor Einar Horsberg Janneche Utne Skaare Kristian Ingebrigtsen 《Biomarkers》2013,18(3):175-180
Juvenile rainbow trout were exposed to unlabelled benzo[a]pyrene BaP and 3H benzo a pyrene (3H BaP), in a static exposure system for 2 days. The initial concentration was 30 μg l-1 and 0.625 μCi l-1, corresponding to 6 mg kg-1 body weight and 125 μCi kg-1 body weight. Hepatic 7-ethoxyresorufin-O-deethylase (EROD) activity was measured during the exposure and depuration periods, elucidating the time course pattern of CYP1A induction. Maximum induction (11-fold) of EROD activity was observed on day 2 after addition of BaP to the water. Tissue distribution of 3H-BaP was studied by liquid scintillation counting and whole body autoradiography. The concentration of 3H-BaP-derived radioactivity was highest in the bile at all sampling times. High levels of radiolabelled compound were also present in the gills, liver and the olfactory organ. There was an overall decrease in all tissues during the depuration period. The elimination of 3H-BaP-derived radioactivity from the gills, however, was slow compared with liver and blood (6.2 days vs 2.7 and 2.9 days, respectively). 相似文献
5.
Degradation of Amino Acids at a Simulated Water-sediment Interface of a Mediterranean Lagoon Environment, Golfe du Lion The degradation of two 14C and 3H labelled amino acids at the simulated water-sediment interface of a Mediterranean lagoon was studied. The four day experiments included the respiratory activity measurement and the study of incorporation processes of the radioactivity in specific organic fractions of the 14C arginine. After active mineralization occurred during the first day, the radioactivity in the acid-soluble fraction decreased and polycondensed products were progressively incorporated. The radioactivity of the 3H lysine was included mainly in the acid soluble fraction. The nature of the substrate, and the reducing conditions in the environment affect the radioactivity distribution in the organic fractions. The identification of amino acids in the acid-soluble and base soluble fractions shows that the labelled arginine and lysine and other amino acids in the acid soluble and base soluble fractions shows that the labelled arginine and lysine released from the initial compounds are important quantitatively. 相似文献
6.
Inhibitory neurons innervating the muscle receptor organ (MRO) of crayfish were used to study the uptake and release of tritiated GABA. MROs that have been directly exposed to 3H GABA for 60–75 min release radioactivity during low-frequency electrical stimulation. When ganglia containing the inhibitory cell bodies are exposed to 3H GABA, the isotope travels along a pathway unique to the inhibitory axon, at rates that range between 160 and 240 mm per day. Electrical stimulation of inhibitory axons whose cell bodies have been exposed to 3H GABA for 4–5 hr produces release of isotope from isolated MROs. Low calcium, high magnesium exposure prevents the stimulus-dependent release of radioactivity. Thin layer chromatographic analyses indicate that GABA comprises at least a major fraction of the radioactivity collected from stimulated preparations. A number of unidentified radioactive compounds are usually present with GABA, and it is suggested that most of these are catabolites of GABA. 相似文献
7.
A procedure for the preparation of highly radioactive homogeneous 32P-labeled 3-hydroxy-3-methylglutaryl coenzyme A reductase from rat liver microsomes has been developed. The enzymatic preparation obtained by this procedure has a specific radioactivity 50-fold higher than that reported in previous literature. The purified enzyme was judged to be homogeneous on the basis of comigration of enzyme activity with a single band of protein and 32P radioactivity on polyacrylamide gels. The 32P covalently bound to the reductase was removed upon incubation with purified hydroxymethylglutaryl coenzyme A reductase phosphatase from rat liver. 相似文献
8.
Summary Auxin activity was detected in honeydew obtained from the aphid Tuberolachnus salignus (Gmelin) feeding on willow (Salix viminalis). Active uptake of 14C-indolyl-3-acetic acid (IAA) into the sieve tubes was demonstrated by irrigating the cambial surface of willow bark with 14C-IAA solution and assaying aphid stylet exudate. When, however, 14C-IAA was applied to the peridermal tissues of the bark or to a mature leaf most of the radioactivity (collected in honeydew or stylet exudate) co-chromatographed with indolyl-3-acetyl-aspartic acid (IAAsp). The presence of IAAsp in honeydew was not affected by extraction procedure or by aphid metabolism. Honeydew obtained from willow treated with 14C-tryptophan contained only 14C-tryptophan. When 14C-IAA was applied in agar to the cut end of willow segments the radioactivity was found to move in a basipetally polar manner. The direction of movement of radioactivity in the sieve tubes, however, was found to be influenced by the proximity of the roots. Nevertheless, there was evidence that endogenous auxin in the sieve tubes does move in a predominantly basipetal direction. 相似文献
9.
[3H] Kainic acid was administered intraventricularly to rats at a dose that selectively destroys the pyramidal cells of hippocampal area CA3. Only about one-third of the injected radioactivity was recovered in the brain 15 min later, but the residual radioactivity disappeared at a much slower rate. [3H]-Kainic acid distributed rather evenly throughout the brain; there was no correlation between accumulation of radioactivity and neurotoxicity. Almost 90% of the radioactivity in sucrose homogenates was recovered in the high-speed supernatant. No cerebral metabolism of [3H] kainic acid was detected by thin-layer chromatography. These data rule out the possibility that a lethal accumulation of the toxin by hippocampus accounts for the preferential vulnerability of hippocampal pyramidal cells. 相似文献
10.
A study of the intracellular transport of calcium in rat heart 总被引:4,自引:0,他引:4
The distribution of in vivo injected 45Ca++ in the subcellular fractions of rat heart has been studied. Most of the radioactivity of the cell was found to be associated with the subcellular organelles; only a small fraction was recovered in the soluble phase. Mitochondria contained the greatest part of the total radioactivity associated with the subcellular organelles. After injection of 45Ca++ the specific activity of the mitochondrial calcium pool was several times higher than that of the calcium of the sarcoplasmic reticulum. Pentachlorophenol has been administered to rats to uncouple oxidative phosphorylation in heart mitochondria in vivo and its effect on the distribution of 45Ca++ in the heart studied. Under these conditions, it has been found that mitochondria contained much less 45Ca++ than the controls; this decrease was paralleled by an increase of the radioactivity associated with the microsomes and with the final supernatant. Experiments in which 45Ca++ was added to heart homogenates at 0° indicated that 45Ca++ also became bound to mitochondria and the other subcellular structures at 0°. However, PCP had no effect on the distribution of radioactivity among the subcellular fractions under these conditions. The results suggest that (1) energy-linked movements of Ca++ take place in mitochondria of the intact rat heart, (2) a part of the uptake of 45Ca++ by mitochondria does not depend on metabolism, and, (3) the movements of Ca++ in heart mitochondria in vivo are probably more active than those in the sarcoplasmic reticulum. 相似文献
11.
Total polysomes from human term placenta were incubated in a wheat germ cell-free system during 1 hr at 25° C. Human placental lactogen hormone was identified among the proteins synthesized in vitro by immune precipitation and subsequent sodium dodecylsulphate electrophoresis of the immunoprecipitate. The zone containing the radioactivity from the immunoprecipitate [3H]-labelled lactogen hormone comigrated exactly with the radioactivity zone from added [14C]-labelled marker hormone. This result indicates that the molecular weight of the synthesized product must be equal or very similar to that of the native protein hormone. 相似文献
12.
[3H]-cocaine, [3H]-norcocaine, [3H]-benzoylecgonine and [3H]-benzoylnorecgonine were administered i.c. in equi-potent pharmacologic doses and the intracellular disposition and metabolism of each drug determined. Norcocaine and cocaine rapidly entered and egressed from the brain so that 4.8–6.1% of the radioactivity present in brain at one minute was observed at 30 minutes. The highest levels of subcellular radioactivity were generally found in the microsomal plus supernatant, followed by the nuclear and shocked mitochondrial fractions. No apparent localization of the radioactivity occured in synaptic membranes. The brain/plasma (B/P) ratio curves for cocaine and norcocaine were similar; however, the norcocaine values were considerably higher at each time interval. Benzoylecgonine and benzoylnorecgonine had higher comparative B/P ratios than cocaine or norcocaine and persisted in brain for a longer period of time so that 0.6–2.1% of the radioactivity present in brain at 1 hour was detected at 24 hours. Cocaine and norcocaine were extensively metabolized to the benzoylmetabolites. Benzoylecgonine was metabolized to benzoylnorecgonine and benzoylnorecgonine was unmetabolized. The brain disposition data and B/P ratios agreed quite well with the overall pharmacologic action of cocaine and its metabolites. 相似文献
13.
《International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology》1987,14(6):587-592
[11C]Cyanoimipramine has been prepared by methylation of the desmethyl cyanoimipramine with [11C]methyl iodide. The chemically and radiochemically pure labelled product was obtained with a high specific activity (> 300 mCi/μmol). When 11C (or 3H)-cyanoimipramine was intravenously administered in mice, high accumulations were shown in brain and lung. Thirty minutes after injection of the tracer, differences were found in the radioactivity between the cerebral cortex and the cerebellum. The regional distribution of radioactivity in the rat brain 30 min after i.v. injection of [11C]cyanoimipramine was also examined, and the radioactivity was high in receptor rich areas (striatum, cerebral cortex etc.) but low in receptor poor area (cerebellum). The in vivo stability of [3H]cyanoimipramine was quite stable in the mouse brain for at least 30 min. Thirty minutes after injection, the radioactivity in the cerebral cortex of the carrier-added state was reduced as compared with the carrier-free state. Taken together, the in vivo specific binding of [3H]cyanoimipramine in the cerebral cortex was estimated at about 40–50% of the total radioactivity. Furthermore, the distribution of [3H]cyanoimipramine in the mice forced to swim was examined. Significant changes in the distribution of [3H]cyanoimipramine were observed in the cerebral cortex. 相似文献
14.
Donald E. Kizer David P. Ringer Boyd A. Howell 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1983,740(4):402-409
The distribution of radioactivity among pyrimidine isostichs (or isoplyths) of DNA from 24-h regenerating rat liver was studied with [3H]Thd, [14C]orotate or with inorganic 32Pi. Expression of incorporated radioactivity as log10% of total radioactivity recovered for each of the 11 pyrimidine isostichs detected showed that radioactivity from [3H]Thd was asymmetrically distributed among the isostichs, i.e., 3H radioactivity failed to access regions of DNA yielding lower molecular weight pyrimidine isostichs as efficiently as it accessed regions yielding higher molecular weight pyrimidine isostichs. The thymine (T) content of isostichs exceeded that of cytosine (C), i.e.,
ratios for the first 10 isostichs averaged 1.43 ± 0.08 and 1.28 ± 0.05, depending on the method of analysis; furthermore, the
ratio for isostich 1 was significantly higher than ratios for isostichs 2 through 10. Asymmetric distributions of [3H]Thd radioactivity also were seen at 18 or 30 h post-partial hepatectomy. Thus, radioactivity from [3H]Thd, a DNA precursor from the salvage pathway, failed to efficiently access lower molecuar weight isostichs despite thymine enrichment, suggesting that thymine moieties were supplied from additional sources. Radioactivity from [14C]orotate accessed lower molecular weight pyrimidine tracts more efficiently than [3H]Thd, but less efficiently than it accessed higher molecular weight isostichs, resulting in an asymmetric distribution of 14C radioactivity. This result suggested that appreciable quantities of thymine and cytosine moieties utilized for DNA synthesis were supplied de novo, but other sources also were utilized. Radioactivity from 32Pi, a de novo precursor, was distributed symmetrically, i.e., the slope among lower molecular weight isostichs increased enough that it was indistinguishable from slopes for intermediate and higher molecular weight isostichs. Since 32P radioactivity among lower molecular weight isostichs reflects appreciable contributions of de novo phosphate moieties from both pyrimidine- and purine-containing deoxynucleoside triphosphates, opportunities for observing contributions of 32P radioactivity from pathways other than the de novo pathways appeared to lie beyond limits of detectability. The distribution of radioactivity from labeled DNA precursors among lower molecular weight pyrimidine tracts (a) indicate that thymine moieties are contributed by both salvage and de novo pathways; (b) support the possibility that cytosine moieties also are contributed by both pathways; and (c) support the ‘replitase’ concept for channeling dNTPs to replicating forks. 相似文献
15.
Summary Isolated oocytes of Perinereis cultrifera have been incubated in culture media with added [3H]glycerol, [14C]butyric acid or [14C]oleic acid. The principal neutral lipid synthesized was triacylglycerol, although incorporation of radioactivity into other lipid categories (sterol, fatty acid, wax ester) was also observed. A more significant percentage of triacylglycerol was labelled after incubation with [3H]glycerol and [14C]oleic acid than with [14C]butyric acid. With this precursor, monoacylglycerol appears to be the class of lipid compartment which initially show the most radioactivity. Electron microscopic autoradiography has revealed that labelling after incorporation of glycerol was mainly localized on the lipid droplets but not on the yolk granules. A second metabolic pathway is represented by phospholipid membrane synthesis. 相似文献
16.
The ability of juvenile turbot, Scophthalmus maximus (L.), to elongate and desaturate various polyunsaturated fatty acids (PUFA) was examined in relation to their lipid composition. Triacylglycerols were the most abundant lipid class present in the fish and phosphatidylcholine was the predominant phospholipid. In all lipid classes examined the levels of (n-3) PUFA exceeded that of (n-6) PUFA. 18C PUFA were minor components in comparison with 20:5(n-3) and 22:6(n-3). 20:4(n-6) was present in highest concentration in phosphatidylinositol in which it accounted for 16.9% of the fatty acids. When the fish were injected with either 14C-labelled 18:2(n-6), 18:3(n-3), 20:4(n-6), 20:5(n-3) or 22:6(n-3) the highest percentage recovery of radioactivity (69%) in body lipid was observed with 22:6(n-3). With all labelled substrates free fatty acids contained only a small proportion of the total recovered radioactivity whereas triacylglycerols were highly labelled. Phosphatidylcholine/sphingomyelin was the most highly labelled polar lipid fraction. With 14C-20:4(n-6) as injected substrate, 23.2% of the radioactivity recovered in total lipid was present in phosphatidylinositol in comparison with less than 6% with the other substrates. Only small proportions of radioactivity from 14C-18:2(n-6) and 14C-18:3(n-3) were recovered in the 20 and 22C fatty acids of triacylglycerols and total polar lipid. With 14C-20:5(n-3) as substrate, 27 and 33% of the total radioactivity recovered in the fatty acids of triacylglycerols and polar lipids respectively was present in 22C fatty acids. The corresponding values for l4C-20:4(n-6) as substrate were 19 and 18%. The results confirm the limited capacity of turbot to convert 18C PUFA to longer chain PUFA but demonstrate their ability to synthesize 22C PUFA from 20C PUFA. They also suggest a small but specific requirement for 20:4(n-6). 相似文献
17.
The relationship between protein synthesis and the incorporation of [3H]gibberellin A1 ([3H]GA1) into a 2,000xg pelletable (2KP) fraction from lettuce (Lactuca sativa L.) hypocotyl sections has been investigated. Concentrations of D-2-(4-methyl-2,6-dinitroanilino)-N-methylpropionamide (MDMP) between 10-7 M and 10-4 M caused increasing inhibition of growth, 2KP labelling and incorporation of [14C]leucine into soluble protein. Growth and 2KP radioactivity were highly correlated (r=0.996). Transfer to MDMP early or late in the course of GA response caused reductions in both growth and incorporation into the 2KP fraction. Exposure to the inhibitor had more effect at 4 h than at 20 h. The proportions of alkali-soluble and insoluble radioactivity in the 2KP fraction were also altered by this treatment. The implications of these findings are discussed.Abbreviations GA1
gibberellin A1
- MDMP
D-2-(4-methyl-2,6-dinitroanilino)-N-methylpropionamide
- 2KP
a2,000xg pelletable fraction 相似文献
18.
—The electrophoretic patterns of proteins, radioactivity and acetylcholinesterase (AChE, EC 3.1.1.7) extracted by water or Triton X-100 from resting and stimulated sympathetic ganglia were determined by analysis on polyacrylamide gels. A concomitant study by ultrastructural histochemistry showed that the bulk of the acetylcholinesterase activity of the sympathetic neurons and the preganglionic nerves was localized within the endoplasmic reticulum. The AChE molecules had low relative electrophoretic mobilities; they could be resolved into 3 bands: one major and two minor. The radioactivity profile obtained after labelling with l -[U-14C]leucine displayed 3 main peaks which are located at the beginning, the middle and the front of the electrophoretograms. The set of proteins and the peak of radioactivity both of which have migrated at the beginning of the gel, as well as the major band of AChE, could be extracted only in the presence of Triton X-100. The radioactivity present in the region of the acetylcholinesterase activity decreased by about 50% after a chase period of 5-8 h. The preganglionic stimulations at 16/s, lasting 1-4 h, decreased by about 50% the incorporation of l -[U-14C]leucine into proteins which migrated at the beginning and the front of the electrophoretograms. These stimulations reduced by half the radioactivity present in the region of the AChE bands. 相似文献
19.
Summary The reasons underlying the initial increase and subsequent decrease in the amount of radioactivity in the receiver block at the apical end of a Zea root segment supplied with a basal donor block containing labelled IAA have been investigated.The phenomenon was observed in segments supplied with IAA-1-14C, IAA-2-14C and IAA-5-3H. An acropetal polarity in the movement of radioactivity into the receiver blocks was observed using donor blocks containing IAA-5-3H at concentrations as low as 10-10M.The decrease in the amount of radioactivity in the receiver block begins after 6–8 h of transport at 25° C, and is unaffected by renewal of the donor block every 2 h, or the presence of 2% sucrose in the donor and receiver blocks.The net export of radioactivity into the receiver block at the apical end of the segment virtually ceases after 6–8 h of transport at 25° C, and is not prolonged by the presence of 2% sucrose in the donor and receiver blocks. At 10° C, net export of radioactivity continues for at least the first 50 h of transport, and the amount of radioactivity in a continuously applied receiver block continues to increase over this period.Receiver blocks removed from the apical end of segments after 8 h of transport and placed on planchettes show little or no decrease in the amount of radioactivity they contain as a function of time, in marked contrast to those left in contact with the segment.There is a marked, and metabolically dependent, resorption of radioactivity from the receiver block at the apical end of the segment after about 8 h of transport at 25° C; most of the resorbed radioactivity remains in the apical 2–4 mm of the segment.There is a loss of radioactive CO2 from segments supplied with a basal donor block containing 10-6M IAA-1-14C at 25° C, the emission beginning after 6–8 h of transport. Segments similarly supplied with 10-6M IAA-2-14C did not begin to lose radioactive CO2 until after about 10–12 h of transport.The ability of the segments to transport radioactivity in a polar manner declines with time after they are excised from the root, regardless of whether their cut ends are kept in the intervening period in contact with plain agar blocks, or ones containing unlabelled IAA at 10-6M. By the 6th h after excision at 25° C no transport of radioactivity through the segments and into the receiver blocks could be detected in either the aropetal or basipetal direction.The decrease in radioactivity in the receiver block after transport periods of 6–8 h at 25° C is therefore due to (1) a cessation of net export of radioactivity into the block, and (2) the onset of a metabolically-dependent, net resorption of radioactivity. At this time substantial amounts of radioactive CO2 begin to be evolved from segments supplied with IAA-1-14C, whereas with IAA-2-14C radioactive CO2 is not evolved for a further 4–6 h. 相似文献
20.
Peter Adamietz Reinhard Bredehorst Helmuth Hilz 《Biochemical and biophysical research communications》1978,81(4):1377-1383
(3H)poly(ADP-ribose) synthesized from nuclei by incubation with (3H)NAD was released from protein by alkaline treatment and electrophoresed in dodecyl sulfate gels. Individual polymers up to at least 33 units were completely separated according to their chain length. Size distribution was visualized by fluorography of the gels, and quantified by radioactivity determination of sliced gels The method could be applied to crude nuclear extracts. It showed that nuclei of Ehrlich ascites tumor cells produced a poly(ADP-ribose) pattern distinctly different from that of rat liver nuclei. 相似文献