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1.
利用放射性同位素示踪技术研究了14C标记菲在有控系统中的迁移转化。结果表明,14C-菲在有控系统中降解较快,施入药品24d后仅有0.32%的14C-菲存留在植物、营养液和火山石中。植物吸收的14C放射性大部分被结合到植物组织中,其它14C主要以菲的极性代谢物形态存在。  相似文献   

2.
Abstract Monic acid A was isolated from a Pseudomonas fluorescens fermentation in which pseudomonic acid A (PA) was the principal secondary metabolite. [3-14C]3-Hydroxy-3-methyl-glutaric acid (HMG) given early in the idiophase radiolabelled PA (1.1% incorporation), confirming experimentally the putative direct involvement of HMG in the biosynthesis of PA, but contributed relatively insignificant radiolabel to the monic acid extracted from the broth at the end of the fermentation. Ethionine inhibited (80%) PA biosynthesis and correspondingly reduced incorporation of [14C]HMG. In contrast, ethionine increased incorporation of [methyl-14C]methionine into PA and enhanced specific radioactivity of the antibiotic 8-fold. Ethionine inhibition of secondary metabolite methylations did not divert pseudomonate biosynthesis to give unusual analogues, implying that methylation of a putative pentaketide precursor of the monate moiety forms a vital intermediate of the pseudomonate pathway, but caused a new [14C]HMG-derived polar metabolite of biosynthetic interest to become evident.  相似文献   

3.
The irreversible binding of [14C]2,2′-di- and [14C]2,4,5,2′,4′,5′-hexachlorobiphenyl ([14C]DCB and [14C]HCB) to protein was studied in the presence of rat liver microsomes and a NADPH-generating system. Protein-bound radioactivity was found with [14C]DCB but not with [14C]HCB. The binding of 14C-metabolites was increased by pretreatment of the rats with phenobarbital or polychlorinated biphenyls. Protein binding was linear for 80 min. In contrast, monohydroxy-metabolites of DCB were formed and degraded within 40 min. Inhibition of secondary oxidation of DCB by scavening superoxide anions or by glucuronidation of the monophenols markedly decreased the protein binding. Addition of trichloropropene oxide or styrene oxide, both inhibitors of epoxide hydrase, did not significantly stimulate the binding. The results suggest that the majority of reactive metabolites of DCB arise from secondary metabolism, i.e., the subsequent oxidation of the phenolic metabolites. Arene oxides, the primary products, appear to play a minor role in the protein binding of DCB.  相似文献   

4.
Assessing petroleum biodegradation rates is an important part of predicting natural attenuation in subsurface sediments. Monitoring carbon dioxide (CO2) and methane (CH4) produced in situ, and their radiocarbon 14C), stable carbon (13C) and deuterium (D). signature provide a novel method to assess anaerobic microbial processes. Our objectives were to: (1) estimate the rate of anaerobic petroleum hydrocarbon (PH) mineralization by monitoring the production of soil gas CH4 and CO2 in the vadose zone of low-permeability sediment, (2) evaluate the dominant microbial processes using δ13C and δD, and (3) determine the proportion of CH4 and CO2 attributable to anaerobic mineralization of PH using 14C analysis. Argon was sparged into the subsurface to dilute existing CO2 and CH4 concentrations. Vadose zone CO2, CH4, oxygen, total combustible hydrocarbons, and argon concentrations were measured for 75 days. CO2 and CH4 samples were collected on day 86 and analyzed for 14C, δ13C, and δD. Based on CH4 soil gas production, the anaerobic biodegradation rate was estimated between 0.017 to 0.055 mg/kg soil-d. CH4 14C (2.6 pMC), δ13C (-45.64‰), and δD (-316‰) values indicated that fermentation of PH was the sale source of CH4 in the vadose zone. CO2 14C (62 pMC) indicated that approximately 47% of the total CO2 was from PH mineralization and 53% from plant root respiration. Although low-permeability sediment increases the difficulty of completely replacing in situ soil gas and assuring anaerobic conditions, this novel respiration method distinguished between anaerobic processes responsible for PH degradation.  相似文献   

5.
Claudia Kluge  Peter Dimroth   《FEBS letters》1994,340(3):245-248
Subunit c of the F1F0-ATPase from Propionigenium modestum was extracted from the particulate cell fraction with chloroform/methanol. The protein was further purified by carboxymethyl cellulose chromatography and anion exchange HPLC in the organic solvent. SDS-PAGE of the purified protein indicated a single stained protein band migrating as expected for the c-subunit. Incubation of isolated subunit c in chlorform/methanol or aqueous buffer containing dodecyl-β- -maltoside with [14C]dicyclohexylcarbodiimide (DCCD) resulted in the incorporation of radioactivity into the protein. The rate of this reaction depended on the external pH; it was significantly faster in the more acidic than in the alkaline pH range. In the presence of Na+ subunit c was partially protected from labeling with [14C]DCCD at pH 6.1 and at pH 7.5, whereas no protection was evident at pH 5.5. At pH 7.5, the rate of subunit c labeling by [14C]DCCD in the presence of 20 mM NaCl was about 50% lower than in the absence of Na+ ions. The isolated c-subunit therefore apparently retains in part the Na+ binding site which, when occupied, diminishes the reactivity of the protein towards DCCD.  相似文献   

6.
Dextran was synthesized using dextransucrase from Streptococus sanguis 10558 and (F)-[14C]sucrose as substrate to test the possibility that sucrose may be the initial acceptor for glucose. If sucrose is the initial acceptor, then dextran chains should have [14C] fructose in a terminal ‘sucrose’ linkage which can be cleaved under mild conditions. Although incorporation of [14C]fructose into dextran was observed, the label was not released by mild hydrolysis, indicating that sucrose is not the initiator for dextran synthesis. Incorporation of [14C]fructose into dextran might represent its ability to act as an acceptor, as suggested by the isolation of leucrose as a by-product in the reaction.  相似文献   

7.
Steven C. Huber  Gerald E. Edwards   《BBA》1977,462(3):583-602
1. Evidence is presented for high rates of carrier-mediated uptake of pyruvate into the stroma of intact mesophyll chloroplasts of the C4 plant Digitaria sanguinalis, but not the chloroplasts of the C3 plant Spinacea oleracea. Uptake of pyruvate in the dark with the C4 mesophyll chloroplasts was followed using two techniques: uptake of [14C]pyruvate as determined by silicon oil centrifugal filtration and uptake as indicated by absorbance changes at 535 nm (shrinkage/swelling) after addition of 0.1 M pyruvate salts.

2. Uptake of the pyruvate anion by an electrogenic carrier is suggested to be the major mode of transport. Chloroplast swelling was observed in potassium pyruvate plus valinomycin and uptake of [14C]pyruvate was inhibited by membrane-permeant anions. Valinomycin reduced uptake in the absence of external potassium and the inhibition could be reversed by addition of external potassium.

3. Uptake of pyruvic acid (or a pyruvate /OH antiport) is ruled unlikely since [14C]pyruvate uptake was relatively independent of the pH gradient across the envelope and addition of pyruvate to chloroplasts did not result in an alkalization of the medium. The low rate of swelling observed in ammonium pyruvate may be due to non-mediated permeation of pyruvic acid, which is possible only at high pyruvate concentrations.

4. The concentration of pyruvate in the stroma increased with external concentration over the range tested (up to 40 mM) but the concentration ratio (internal/external) was always less than one. The steady-state concentration of [14C]pyruvate in the stroma was dependent on the ionic strength of the medium, with saturation at roughly I = 0.04 M, while accumulation of the membrane-permeant cation tetraphenylmethylphosphonium decreased with increasing ionic strength. This suggests that ionic strength modifies a membrane potential (inside negative) across the envelope and that pyruvate uptake responds to the magnitude and direction of that potential (−80 mV at low ionic strength).

5. Chloride and inorganic phosphate were potent inhibitors of [14C]pyruvate uptake. Of the sulfhydryl reagents tested, N-ethylmaleimide was not inhibitory while mersalyl completely blocked [14C]pyruvate uptake and swelling in potassium pyruvate plus valinomycin. Pyruvate uptake, as measured by valinomycin induced swelling in potassium pyruvate, was highly temperature sensitive, with an energy of activation of 39 kcal/mol above 9 °C.

6. Phenylpyruvate, -ketoisovalerate, -ketoisocaproate, -cyano-4-hydroxycinnamic acid and -cyanocinnamic acid inhibited [14C]pyruvate but not [14C]-acetate uptake in the dark and also reduced pyruvate metabolism by the chloroplasts in the light.  相似文献   


8.
Uridine and cytidine are major nucleosides and are produced as catabolites of pyrimidine nucleotides. To study the metabolic fates and role of these nucleosides in plants, we have performed pulse (2 h) and chase (12 h) experiments with [2-14C]uridine and [2-14C]cytidine and determined the activities of some related enzymes using tubers and fully expanded leaves from 10-week-old potato plants ( Solanum tuberosum L.). In tubers, more than 94% of exogenously supplied [2-14C]uridine and [2-14C]cytidine was converted to pyrimidine nucleotides and RNA during 2-h pulse, and radioactivity in these salvage products still remained at 12 h after the chase. Little degradation of pyrimidine was found. A similar pyrimidine salvage was operative in leaves, although more than 20% of the radioactivity from [2-14C]uridine and [2-14C]cytidine was released as 14CO2 during the chase. Enzyme profile data show that uridine/cytidine kinase (EC 2.7.1.48) activity is higher in tubers than in leaves, but uridine nucleosidase (EC 3.2.2.3) activity was higher in leaves. In leaves, radioactivity from [U-14C]uracil was incorporated into β-ureidopropionic acid, CO2, β-alanine, pantothenic acid and several common amino acids. Our results suggest two functions of uridine and cytidine metabolism in leaves; these nucleosides are not only substrates for the classical pyrimidine salvage pathways but also starting materials for the biosynthesis of β-alanine. Subsequently, some β-alanine units are utilized for the synthesis of pantothenic acid in potato leaves.  相似文献   

9.
Metabolism of indole-3-acetic acid in soybean [ Glycine max (L.) Merr.] was investigated with [1-14C]- and [2-14C]-indole-3-acetic acid (IAA) applied by injection into soybean hypocotyl sections and by incubation with soybean callus. Free IAA and its metabolites were extracted with 80% methanol and separated by high performance liquid chromatography with [3H]-IAA as an internal standard. Metabolism of IAA in soybean callus was much greater than that in tobacco ( Nicotiana tabacum L.) callus used for comparison. High performance liquid chromatography of soybean extracts showed at least 10 metabolite peaks including both decarboxylated and undecarboxylated products. A major unstable decarboxylated metabolite was purified. [14C]-indole-3-methanol (IM) was three times more efficient than [2-14C]-IAA as substrate for producing this metabolite. It was hydrolyzable by β-glucosidase (EC 3.2.1.21), yielding an indole and D-glucose. The indole possessed characteristics of authentic IM. Thus, the metabolite is tentatively identified as indole-3-methanol-β-D-glucopyranoside. The results suggest that soybean tissues are capable of oxidizing IAA via the decarboxylative pathway with indole-3-methanol-glucoside as a major product. The high rate of metabolism of IAA may be related to the observed growth of soybean callus with high concentrations of IAA in the culture medium.  相似文献   

10.
Abstract: To determine if lactate is produced during aerobic metabolism in peripheral nerve, we incubated pieces of rabbit vagus nerve in oxygenated solution containing d -[U-14C]glucose while stimulating electrically. After 30 min, nearly all the radioactivity in metabolites in the nerve was in lactate, glucose 6-phosphate, glutamate, and aspartate. Much lactate was released to the bath: 8.2 pmol (µg dry wt)−1 from the exogenous glucose and 14.2 pmol (µg dry wt)−1 from endogenous substrates. Lactate release was not increased when bath P o 2 was decreased, indicating that it did not come from anoxic tissue. When the bath contained [U-14C]lactate at a total concentration of 2.13 m M and 1 m M glucose, 14C was incorporated in CO2 and glutamate. The initial rate of formation of CO2 from bath lactate was more rapid than its formation from bath glucose. The results are most readily explained by the hypothesis that has been proposed for brain tissue in which glial cells supply lactate to neurons.  相似文献   

11.
Abstract— The characteristics of the uptake of l -[U-14C] glutamate into rat dorsal sensory ganglia were investigated. The uptake was mediated by two distinct kinetic systems, with apparent Km values of the order of 10−3 M (low affinity) and 10−5 m (high affinity). The high affinity uptake system was strongly dependent upon temperature and sodium ion concn, and was depressed by a number of metabolic inhibitors. Following uptake, [14C] glutamate was extensively metabolized, primarily to glutamine, although this was not so with cultured ganglia, where in addition to an increased uptake of [14C] glutamate, the specific radioactivity of glutamate was increased and that of glutamine decreased. The labelled substrates [U-14C]pyruvate and [U-14C] acetate were used to investigate this phenomenon and the results are discussed in relation to current knowledge of metabolic compartmentation in nervous tissue.  相似文献   

12.
研究了14C-涕灭威在5种土壤中(4ppm,1.22μCi·50g-1 土壤干重)的生物降解。模拟试验为密闭系统,土壤中水分含量为22%,气温20-30℃在供试的5种土壤中,北京肖家河的土壤降解最快,为施人放射剂量的51.3%,以“14CO2 形式从土壤进出;26.0%与土壤结合,只有21.6%可以被抽出。取自浙江义乌的土壤降解较慢,收集到的14CO2 为施入量的23.3%.土壤中加入杀菌剂红霉素或敌茵丹降解作用明显减慢。土壤提取物中涕灭威亚砜、涕灭威亚砜肟被确认是主要的代谢产物,还发现了少量的涕灭威砜,涕灭威亚砜腈涕灭威砜腈和涕灭威砜肟等降解物。  相似文献   

13.
通过测定上海市青浦区东风港百慕大、白花三叶草、高羊茅和白茅等4种典型滨岸草本植物各组织以及不同垂直深度土壤有机质δ13C值,对滨岸草地生态系统的植物-土壤碳稳定同位素特征进行了分析.结果表明: 白花三叶草、高羊茅属于C3植物,百慕大、白茅属于C4植物,其茎叶、凋落物和根系各组织间δ13C值无显著差异.C3和C4植物样带表层土壤有机质δ13C值随着土壤深度递增而呈现截然不同的变化特征,这与样带本底δ13C值以及碳稳定同位素分馏效应有关,同时还受植物根系分布深度的影响.植物输入是土壤有机碳(SOC)的最主要来源,植物有机体δ13C组成对土壤有机质δ13C值有直接影响,植物各组分δ13C值与土壤有机质δ13C值均存在极显著相关.4种草本植物样带SOC含量与δ13C值均呈极显著相关,其中,C3植物样带SOC含量与δ13C值呈线性负相关,C4植物样带SOC含量与δ13C值呈线性正相关.  相似文献   

14.
Previous studies have shown addition of light liquid paraffin to enhance the elimination of organochlorine xenobiotics. In the present study the effect of paraffin on the elimination of [14C]hexachlorobenzene (HCB) was compared with the effect of possible alternative compounds, squalane and sucrose polyester (SPE). Four groups of 7 rats were fed a diet containing 1.5 ppm [14C]HCB for 4 days followed by 10 days on HCB-free diet. Thereafter one group (control) remained on this diet whereas the other 3 groups received a diet supplemented with 8% (w/w) paraffin, squalane or SPE, respectively. Radioactivity in urine and faeces was measured daily and at the end of the experiment in samples of abdominal fat, muscle, liver, kidney and blood. Dietary treatment with either paraffin, squalane or SPE markedly enhanced faecal excretion of [14C]HCB, whereas urinary excretion was not affected. Both the time course as well as the extent of faecal [14C]HCB elimination were similar in the treated groups. After 3 weeks of treatment the amount of [14C]HCB excreted with faeces was about three times higher in treated animals than in controls. The half-life (t1/2) of [14C]HCB elimination from the body was markedly decreased in treated animals (mean 34–38 days) compared to controls (110 days). [14C]HCB concentrations in some major tissues were significantly reduced to the same extent by all three dietary regimens. Thus squalane and SPE are as effective as paraffin in removing HCB from contaminated animals.  相似文献   

15.
A procedure, based on measurement of the stable carbon isotope 13C, has been developed for determining the extent to which petroleum carbon is incorporated into soil organic matter (SOM) by humification of biomass produced during biodegradation of the petroleum in soil. We have shown that a crude oil having a δ13C of-27.4%, when biodegraded in a soil containing SOM with a δ13C of-15.7%, resulted in a change of the δ13C of the bound SOM reflecting that of petroleum carbon. Comparison of five soil biodegradation tests using different amounts and types of fertilizer to stimulate biodegradation of the oil in this soil showed that the extent of the δ13C change in the bound SOM varied with the extent of oil biodegradation observed. To obtain 13C data on the SOM, the residual petroleum was first removed by rigorous extraction with dichloromethane using a Soxhlet apparatus. The extracted soil was then combusted to release bound carbon as CO2, which was analyzed for 13C. Where the SOM has a δ13C similar to that of petroleum, 14C measurements of SOM would give similar results. This type of data, referred to as the petroleum “footprint” in the SOM, could be useful in identifying or confirming intrinsic biodegradation of petroleum in contaminated soil.  相似文献   

16.
In neuroblastoma × glioma hybrid cells (NG 108-15) labelled with [32P]-trisodium phosphate, [3H]-inositol and [14C]-arachidonic acid, bradykinin stimulated the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) while it had no effect on the release of [14C]-arachidonic acid (AA). The effect on PIP, was time- and dose-dependent with a maximal effect on [3H]-inositol- and [32P]-labelled cells after 10–30 s of stimulation with 10−6 M bradykinin. However, the hydrolysis of [14C]-AA labelled PIP2 was delayed compared to the effect on [3H]- and [14C]-PIP2 and was not detectable until after 60 s of stimulation. Bradykinin stimulation resulted in an increased formation of [3H]-inositol phosphates (IP) and [32P]- and [14P]- and [14C]-phosphatidic acid (PA) but the time course for PA formation did not allow the time-course for PIP2 hydrolysis. A reduced labelling of [23P]- and [14C]-phosphatidylcholine was also found in stimulated cells suggesting that PA may derive from other sources than PIP2. In conclusion, our results indicate that bradykinin activates phospholipase C, but not phospholipase A2, in NG 108-15 cells.  相似文献   

17.
以亚热带红壤丘陵区典型水田和旱地土壤为研究对象,向土壤中添加14C标记稻草,培养30 d后,提取与原位土壤中结构相似的14C可溶性有机碳(DOC);将14C DOC加入水田和旱地土壤中,并设置45%、60%、75%、90%和105%田间持水量(WHC)5个水分梯度,在标准状态下(25 ℃)培养100 d,监测14C DOC在土壤中的矿化过程.结果表明: 培养100 d后,两种土壤中28.7%~61.4%的标记DOC被矿化为CO2,且5个水分条件下,水田土壤DOC的矿化率均显著高于旱地,这主要是由于水田土壤DOC的结构组成比旱地土壤更简单.好气条件(56%~75%WHC)有利于两种土壤DOC的分解,淹水条件(105%WHC)则有利于DOC的积累.土壤处于好气条件(45%~90%WHC)时,DOC的生物可分解率及易分解态所占比例均随着含水量的增加而增加.100 d内,水田和旱地易分解态DOC分别占其累积矿化量的80.5%~91.1%和66.3%~72.4%,说明DOC的生物可分解率主要由其易分解态组分所占比例决定.  相似文献   

18.
Abstract: Chains of lumbar sympathetic ganglia from 15-day-old chicken embryos were incubated for 4 h at 36°C in a bicarbonate-buffered salt solution equilibrated with 5% CO2-95% O2. Glucose (1–10 m M ), lactate (1–10 m M ), [U-14C]glucose, [1-14C]glucose, [6-14C]glucose, and [U-14C]lactate were added as needed. 14CO2 output was measured continuously by counting the radioactivity in gas that had passed through the incubation chamber. Lactate reduced the output of CO2 from [U-14C]glucose, and glucose reduced that from [U-14C]lactate. When using uniformly labeled substrates in the presence of 5.5 m M glucose, the output of CO2 from lactate exceeded that from glucose when the lactate concentration was >2 m M . The combined outputs at each concentration tested were greater than those from either substrate alone. The 14CO2 output from [1-14C]glucose always exceeded that from [6-14C]glucose, indicating activity of the hexose monophosphate shunt. Lactate reduced both of these outputs, with the maximum difference between them during incubation remaining constant as the lactate concentration was increased, suggesting that lactate may not affect the shunt. Modeling revealed many details of lactate metabolism as a function of its concentration. Addition of a blood-brain barrier to the model suggested that lactate can be a significant metabolite for brain during hyperlactemia, especially at the high levels reached physiologically during exercise.  相似文献   

19.
In order to specify the source of locally synthesized prostaglandin (PG) E2 which is able to saturate the large class of low affinity PGE2 receptors in chick spinal cord, bioconversion of [1-14C]arachidonic acid into prostanoids was studied in homogenates of chick spinal cord and meninges first without addition of exogenous glutathione (GSH). Homogenates of spinal cord produced 14C-labeled PGE2, PGD2 and PGF2. Homogenates of meninges accumulated much larger amounts of [14C]PGE2 than spinal cord and surprisingly a 14C-labeled arachidonate metabolite referred to as compound Y. Compound Y generation, which was inhibited by indomethacin and enhanced by esculetin, was therefore mediated through the cyclooxygenase pathway. The fact that no labeled compound Y was detected in homogenates incubated with [3H]PGD2 or [3H]PGE2 indicated that compound Y was not degradation product of PGs. Secondly, after addition of exogenous GSH, 14C-labeled compound Y was totally converted into [14C]PGE2. The compound Y which is converted into PGFs after a strong reduction with NaBH4 and into PGE2 after a mild reduction with GSH-hemin system or SnCl2 was therefore assumed to be a 15 hydroperoxy-PGE2 (15 HP-PGE2). These results suggest that PGE2 can be synthesized in meninges either by the classical isomerization of PGH2 or by isomerization of PGG2 followed by a GSH-sensitive reaction.  相似文献   

20.
A shallow, RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine)-contaminated aquifer at Naval Submarine Base Bangor has been characterized as predominantly manganese-reducing, anoxic with local pockets of oxic conditions. The potential contribution of microbial RDX degradation to localized decreases observed in aquifer RDX concentrations was assessed in sediment microcosms amended with [U-14C] RDX. Greater than 85% mineralization of 14C-RDX to 14CO2 was observed in aquifer sediment microcosms under native, manganese-reducing, anoxic conditions. Significant increases in the mineralization of 14C-RDX to 14CO2 were observed in anoxic microcosms under NO3-amended or Mn(IV)-amended conditions. No evidence of 14C-RDX biodegradation was observed under oxic conditions. These results indicate that microbial degradation of RDX may contribute to natural attenuation of RDX in manganese-reducing aquifer systems.  相似文献   

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