The interaction of phosphine with purified cytochrome c and cytochrome oxidase was studied by absorption and circular dichroic spectrometry. Treatment of cytochrome oxidase with phosphine yielded absorption spectra in both the visible and Soret region that were similar to that produced by dithionite. Striking changes in the circular dichroic spectra were also recorded, sugesting that a valence change of the heme iron was accompanied by a conformation change of the prosthetic group.Phosphine treatment of cytochrome c also yielded spectra similar to those produced by reduction with dithionite suggesting a valence change of the heme iron. However, both the time necessary for complete reduction and the concentration of phosphine necessary for this effect suggest that, in comparison with cytochrome oxidase, cytochrome c was relatively insensitive. Reduction of cytochrome oxidase by phosphine had only a very minor effect on the intrinsic dichroic spectrum suggesting little effect on the polypeptide chains.The persistence of a component after reduction by phosphine and the subsequent reduction of this component by dithionite suggests that phosphine primarily reduced only one comonent of the cytochrome aa3 complex. The failure of EDTA and fluoride, which are known ligands of cytochrome a3 to affect the reduction of cytochrome aa3 by phosphine, as well as the occurrence of a 436-nm isosbestic point of the ferric-ferrous absorption spectra suggests that phosphine reduced primarily cytochrome a. However, it is argued that a slow reduction of cytochrome a3 cannot be ruled out. 相似文献
Tumor necrosis factor (TNF) is a key regulator of inflammatory responses and has been implicated in many pathological conditions. We used structure-based design to engineer variant TNF proteins that rapidly form heterotrimers with native TNF to give complexes that neither bind to nor stimulate signaling through TNF receptors. Thus, TNF is inactivated by sequestration. Dominant-negative TNFs represent a possible approach to anti-inflammatory biotherapeutics, and experiments in animal models show that the strategy can attenuate TNF-mediated pathology. Similar rational design could be used to engineer inhibitors of additional TNF superfamily cytokines as well as other multimeric ligands. 相似文献
Accurate early antemortem diagnosis of tuberculosis in dromedary camels is difficult due to the lack of reliable diagnostic test. The present study aimed to evaluate a lateral flow assay-based kit (rapid assay kit) in tuberculosis diagnosis that employs immuno-chromatographic detection of antibodies in serum, plasma, or whole blood. In a dromedary camel herd comprising 337 animals located at Bikaner, Rajasthan, India, 50 adult weak camels (11 males and 39 females) were tested by applying a single intradermal tuberculin test (SIDT) and rapid assay kit. A total of 14 animals (2 males, 12 females) were found positive in rapid assay. In SIDT, four animals revealed a positive reaction in the neck region and seven animals in the tail base. Another male animal was found SIDT positive but negative in rapid assay; it died after 12 months. Nine rapid assay positive animals died asymptomatically in 1- to 11-month period revealing postmortem tuberculosis lesions that were confirmed by Ziehl-Neelsen staining and histopathology. No tuberculous lesion was evident in the animal found positive in SIDT alone. Results of the present study indicated that serological tests like rapid assay kit can serve as a reliable test for antemortem diagnosis of tuberculosis in dromedary camel.
The effects of phosphine on electron transport and on some partial reactions of oxidative phosphorylation of mitochondria from mouse liver, housefly flight muscles and granary weevils has been studied. Phosphine was a strong inhibitor of respiration of mitochondria in the “active” state (state 3), uncoupled state, and ion-pumping state on glutamate, pyruvate plus malate, succinate, α-glycerophosphate, and ascorbate-cytochrome c as substrates. Respiration of mitochondria in state 3 was completely inhibited by about 250 μM phosphine. By contrast, the respiration of mitochondria in state 4 was much less sensitive. This inhibition could not be released by uncouplers suggesting that it is due to a direct effect on electron transport. Only site III was inhibited to any significant extent. Kinetic studies show that the inhibition was noncompetitive with Ki ranging from 1.6×10?5 to 7.2×10?5 depending on the source and purity of cytochrome oxidase. The inhibition of site III was also more pronounced in sonicated particles than in intact mitochrondria. The significance of this is discussed in relation to membrane sideness and topology of the components of the respiratory chain.Phosphine was unable to activate the “latent” ATPase nor did it have any inhibition of the Mg2+-simulated ATPase and only high levels (1.1 mM) showed modest inhibition (41%) of uncoupler-stimulated ATPase. Phosphine had no effect on the ATP-Pi exchange and on the ATP-ADP exchange reaction at concentrations causing strong respiratory inhibition. 相似文献
A rapid, sensitive and reliable mixed indicator paper strip impregnated with dimethyl yellow (0.05%), cresol red (0.1%) and mercuric chloride (1.0%) in methanol has been developed for the detection of phosphine (PH3) in air at the permissible level (TLV). The paper strip turns red in the presence of phosphine. It is highly sensitive and has better shelf-life than indicator strips impregnated with dimethyl yellow plus mercuric chloride or cresol red plus mercuric chloride. 相似文献
Five species of stored-product insects, [Tribolium castaneum (Herbst), T. confusum (DuVal), Rhyzopertha dominica (F.), Sitophilus oryzae (L.) and S. granarius (L.)], exposed to phosphine in nitrogen and various oxygen-deficient atmospheres, showed reduced susceptibility to three concentrations of phosphine (0.02, 0.04 and 0.2 mg litre?1). The last two species also showed reduced susceptibility to phosphine at 2.0 mg litre?1. Generally mortality increased progressively as the level of oxygen was raised. There was a close relationship between the ability of insects to withstand anoxia and survival from phosphine treatment. 相似文献
Inter‐simple sequence repeats (ISSR) polymorphism was used to determine genetic relationships among 75 Sesamum indicum L. accessions of Korean and exotic sesame. Fourteen reliable ISSR primers were selected for the assessment of genetic diversity, yielding 79 amplification products. Of these polymerase chain reaction products, 33% revealed polymorphism among the 75 accessions. Genetic distances ranged from 0 to 0.255, with a mean genetic distance of 0.0687. The 75 accessions were divided into seven groups on the basis of unweighted pair‐group method with arithmetic averages (UPGMA) cluster analysis. The largest group consisted of 25 Korean cultivars, eight Korean breeding lines and 17 world‐wide accessions. The other groups included 25 accessions, several of which contained useful traits. The dendrogram did not indicate any clear division among sesame accessions based on their geographical origin. However, all Korean sesame cultivars except ‘Namsankkae’ were clustered in the same group, indicating a narrow gene pool. Some of the Korean breeding lines were spread along the dendrogram, showing enlargement of genetic diversity. The genetic diversity data uncovered in this study can be used in future breeding programmes. 相似文献