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1.
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   

2.
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   

3.
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   

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.
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.
Preparations of spinach chloroplasts, essentially free from contamination by other cellular material or whole cells, incorporated 14C almost entirely into glycolate, a polyglucan (probably starch) and intermediates of the Calvin cycle and starch synthesis. About 70% of the 14C was found in dihydroxyacetone phosphate, 3-phosphoglycerate and glycolate. Only small amounts were found in sucrose and amino acids.

On the other hand, preparations consisting of particles containing chloroplasts surrounded by a membrane-bound cytoplasmic layer including mitochondria and microbodies, gave a much broader spectrum of 14C-labelled products. Much less of the 14C was found in dihydroxyacetone phosphate and 3-phosphoglycerate. Instead, sucrose, malate, aspartate, alanine and some other amino acids contained about 40% of the 14C incorporated. It is concluded that sucrose is synthesized by cooperation between the chloroplast and the surrounding layer.  相似文献   


8.
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   

9.
化石燃料的大量使用和森林的过度砍伐,引起大气中CO2浓度的大幅度增加,同时由于Suess效应,大气CO2中的δ13C在不断地下降。植物中δ13C的变化是大气CO2浓度和同位素比值变化的敏感指示器。文中利用树木年轮δ13C序列和植物碳同位素分馏模型,尝试恢复了新疆阿勒泰地区近440年来大气δ13C的变化。结果表明,1850年之前,从树木年轮δ13C序列恢复的大气δ13C相对恒定在-6.60‰(R2=0.052),而1850年之后,该大气δ13C明显降低(R2=0.65),平均约为-7.04‰,平均年降低0.0084‰。这一结果高于从冰芯气泡所恢复的大气δ13C,1850年~1981年冰芯大气δ13C平均年降低约0.00657‰这可能与从树木年轮δ13C序列恢复的大气δ13C有更高的分辨率及树木生长点大气δ13C不同于全球大气δ13C值有关。  相似文献   

10.
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   


11.
为探究高海拔地区的植物碳(C)循环过程与其生境的关系,以生长在高山地区的豆科灌木鬼箭锦鸡儿为研究对象,沿着横跨我国东西部山区的样带采集35个样点的鬼箭锦鸡儿叶片和土壤样品,分析了鬼箭锦鸡儿叶片碳稳定同位素组成(δ13C)、土壤δ13C、叶片和土壤δ13C差值(Δδ13C)在不同采样点的特征及其与气候因子、叶片和土壤元素的关系。结果表明:鬼箭锦鸡儿叶片δ13C的变化范围为-30.9‰~-27.1‰,平均值为-28.4‰,土壤δ13C的变化范围为-26.2‰~-23.2‰,平均值为-25.3‰,Δδ13C的变化范围为2.0‰~7.7‰,平均值为3.1‰;叶片δ13C显著低于土壤δ13C,且随着叶片δ13C增加,土壤δ13C先降低后升高;叶片δ13C与生长季均温和叶片C含量呈显著负相关,土壤δ13C与相对湿度和最暖月均温呈显著负相关,与土壤碳∶氮(C∶N)呈显著正相关,随土壤C含量的增加土壤δ13C先降低后升高,Δδ13C与叶片C含量、土壤C含量和土壤C∶N呈显著正相关;气候因子对叶片δ13C和Δδ13C具有直接影响,同时也通过对叶片和土壤元素的影响,间接导致叶片δ13C、土壤δ13C和Δδ13C的改变。高海拔地区的气候因子、叶片和土壤元素共同影响鬼箭锦鸡儿的C循环过程。  相似文献   

12.
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   

13.
聚四氟乙烯(PTFE)塑料管研磨法是测定植物碳同位素比率(δ13C)值常用的前处理方法。该方法处理样品高效快捷,但对植物δ13C可能存在污染。本研究利用人工气候室开展双因素交互试验,包括空气相对湿度(50%和80%)和空气δ13C(13C富集和贫化的空气)两个因素,对比了PTFE塑料管研磨法和不锈钢管研磨法处理C4植物糙隐子草δ13C的结果。结果表明: 在相同湿度条件下,不同空气δ13C处理的植物13C分馏值(Δ13C,矫正了光合作用底物的δ13C差异)原本可以视为重复,但由于PTFE塑料颗粒的混入,相同湿度不同13C丰度空气培养下植物叶片Δ13C平均差值为4.8‰。该污染效应导致单个叶片δ13C测定的误差高达8‰。考虑到C4植物的Δ13C较低(通常为1‰~8‰),这种污染效应已经超出了可以接受的误差范围。通过建立类似Keeling曲线的二元混合模型对误差进行了有效消除,并准确估算了植物样品和污染物的δ13C。说明广泛采用的PTFE管研磨方法对研究C4植物Δ13C并不适用,将导致较大的误差。对精度要求较高的研究内容建议使用不锈钢瓶进行研磨。  相似文献   

14.
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   

15.
生长在超干旱环境下的3种相思树种表现出异常低的叶片、树枝、树干、根中δ13C含量 在植物生理生态学中,叶片中碳13(13C)含量负值较少(富集),表明叶片处于通过气孔的气体交换减少,比如在干旱胁迫下。此外,与叶片相比,13C在非光合组织中的负值也较少。然而,对从叶片(光合器官)到树枝、树干和根(非光合器官)中的δ 13C数值的关系知之甚少,特别是缺少在关联密切的多个树种间或者不同器官间,以及对生长在极端高温和干旱胁迫下的树木中进行测定。本研究测定了3种近缘沙漠相思树种(Acacia tortilis、A. raddiana和A. pachyceras)从叶片到根的13C含量。我们在以色列南部成树的叶片组织中测定了δ 13C含量。与此同时,在试验果园进行了为期7年的3个水平的灌溉试验。在试验结束时,测定了叶片、树枝、树干和根的生长参数和δ 13C含量。研究结果表明,叶片组织中δ 13C含量约为−27‰,其同位素贫化程度远超过生长在地球上最干燥和最热环境中的沙漠树种的预期值。在不同的相思树种和不同器官中,所有灌溉水平处理中的δ 13C含量并没有富集(−28‰到ca. −27‰),证实了在成熟相思树中的测定结果。在不同器官中,叶片δ 13C含量与树枝和根的δ 13C含量异常相似,甚至比树干的δ 13C含量负值更少。高度贫化的叶片δ 13C表明,尽管这些树木生长在极端干燥的生境中,但其气孔气体交换较高。非光合组织中缺乏δ 13C富集可能与叶片和异养组织生长的季节耦合有关。  相似文献   

16.
Light microscope autoradiography was used to study the retrograde transport of labelled material after injection of [3H]serotonin ([3H]5-HT), [3H]5-hydroxytryptophan ([3H]5-HTP) and [14C]5-hydroxyindoleacetic acid ([14C]5-HIAA) into the olfactory bulb (OB) of rat. A perikaryal labelling was clearly visualized in the Raphe Dorsalis (RD) and the Raphe Centralis (RC) 24 h after injection of [3H]5-HT (but not after injection of [3H]5-HTP or [14C]5-HIAA) into the OB of rats without monoamine-oxidase inhibitor (MAOI). In the OB, the labelled cells (mitral, granular, periglomerular and tufted cells) and the varicosities (dispersed in granular, plexiform and glomerular layers) were greater in number and intensity at 8 h than at 24 h after [3H]5-HT (10−3 M) injection. Five hours after injection of [14C]5-HIAA (10−3 M) some mitral, granular and tufted cells were labelled in the cytoplasm, nuclei and dendrites. A few varicosities were also observed. In contrast, after [3H]5-HTP injection no clear labelling was visualized in axonal processes. A net autoradiographic reaction was seen, however, in the capillary walls and some granular cells.

After injection of [3H]5-HT at various concentrations (10−2 M to 10−5 M) into the OB of rats pretreated with MAOI, a selectivity in the pattern of labelling in the injection site and the afferent cell bodies was found at 10−4 M and 10−5 M. At these concentrations, the serotoninergic RD and RC neurons were clearly labelled, but the non-serotoninergic neurons such as those originating in the Locus Coeruleus, prepiriform cortex were devoid of label. In the OB, only varicosities and fiber-like structures were reactive. In the RD cell bodies, the intensity of labelling as well as the number of labelled cells were greater at higher concentrations of injected [3H]5-HT and when rats were pretreated with a MAOI.  相似文献   


17.
稳定碳同位素组成能精确指示生态系统碳循环过程,可以为深入研究森林演替进程对碳循环过程和固碳潜力的影响提供关键信息.利用稳定碳同位素技术对长白山阔叶红松林演替序列3种林分——中龄杨桦次生林、成熟杨桦次生林、阔叶红松林的叶片、树干、根系、凋落物和土壤δ13C值及碳、氮元素含量进行测定.结果表明: 各演替序列优势树种叶片δ13C从冠上到冠下均呈降低趋势;树干δ13C表现为树皮小于木质部;根系δ13C表现为细根小于粗根.阔叶红松林未分解凋落物δ13C小于半分解及全分解凋落物,次生林相反;土壤δ13C沿深度逐渐增加.总体上,δ13C值叶片<凋落物<根系<树干<土壤,说明植物各器官之间有明显的碳同位素分馏效应,且相同器官不同部位之间也存在差异;植物δ13C沿演替方向先减小后增加,土壤δ13C沿演替方向不断增加,且变化规律可以通过氮元素含量与碳同位素分馏效应的关系解释,说明长白山阔叶红松林演替过程优势树种和碳周转速率的变化影响了碳同位素分馏.  相似文献   

18.
Interaction of mitochondrial F1-ATPase with the isolated natural inhibitor protein resulting in the inhibition of multi-site ATP hydrolysis is accompanied by the loss of activity at low ATP concentrations when single-site hydrolysis should occur. Catalytic site occupancy by [14C]nucleotides in F1-ATPase during steady-state [14C]ATP hydrolysis, which is saturated in parallel with single-site catalysis, is prevented after blocking the enzyme with the inhibitor protein.  相似文献   

19.
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   

20.
特定化合物同位素分析(CSIA)可以实现对复杂基质中特定化合物稳定碳同位素组成(δ13C)的精确测定。应用此方法测定树木非结构性碳水化合物(NSC)中特定成分(如糖类、有机酸和糖醇)的δ13C,不仅能够追踪新同化的光合产物在树木中的运移及与外界的碳交换,还能够更敏感地指示树木生理状况对环境变化的响应。本文首先系统介绍了CSIA从样品采集、处理到δ13C测定的方法,然后综述了树木NSC中各成分之间及各成分在不同器官之间的δ13C差异,阐述了树木NSC的δ13C时间动态变化特征及内在机制,最后分析了NSC作为主要呼吸底物,其δ13C与树木呼吸释放CO2的δ13C(δ13CR)之间的联系,并针对CSIA分析技术在后光合分馏、树木逆境生理和年轮δ13C形成机制等研究的应用前景提出了展望。  相似文献   

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