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1.
The direct and indirect methods for assaying the superoxide dismutase activity of a compound are compared. With the use of a direct method. the mechanism of the catalysis of O2-dismutation by the tested compound can be determined. while with the indirect method it cannot. and this may lead to misinterpretation of the results. Assuming that the catalysis occurs via the ‘ping-pong’ mechanism, both the direct and indirect methods are limited to the determination of values of kcat ≥ 105M?1s?1 and kcat ≥ 3 × 106M?1s?1. respectively. Moreover, many side reactions may occur with the indirect method which may interfere with the measurements. Nevertheless. the indirect method approximates better the in vivo conditions than the direct method, and a tested compound that has high SOD activity using a direct method and low SOD activity using an indirect method. will most probably be a poor SOD mimic in vivo.  相似文献   

2.
Methanobacterium thermoautotrophicum, growing on medium supplemented with 2 mol 63NiCl2/l, was found to take up 1.2 mol 63Ni per g cells (dry weight). More than 70% of the radioisotope was incorporated into a compound, which dissociated from the protein fraction after heat treatment, was soluble in 70% acetone, and could be purified by chromatography on QAE-Sephadex A-25, Sephadex G-25, and DEAE cellulose. The purified 63Ni labelled compound had an absorption spectrum and properties identical to those of factor F 430 and is therefore considered to be identical with factor F 430.Factor F 430, a compound of molecular weight higher than 1000 with an absorbance maximum at 430 nm, has recently been purified from Methanobacterium thermoautotrophicum (Gunsalus and Wolfe, 1978). The structure and function of this compound are not yet known.  相似文献   

3.
NADH-dependent reduction of polyvanadate was observed by using rat liver microsomes as the enzyme source. The reduced vanadate form obtained was blue in color with a broad absorption maximum in the red region around 650 nm. Microsomes and phosphate anions were found to be essential for polyvanadate reduction. The rate and the extent of formation of blue color compound was dependent on the amount of vanadate present. Cytochrome b 5 was found to be involved in this SOD-insensitive reaction. The rate of disappearance of the blue-colored compound was dependent on concentration of NADH and was found to be sensitive to SOD. Catalase and Mn2+. which inhibit oxygen consumption accompanying NADH oxidation, increased both the rate and extent of the blue color compound formed. The results suggest that vanadate acts as an electron acceptor.  相似文献   

4.
The relaxation rates (1 / T 1 and 1 / T 2) in cysts have already been analyzed in terms of materials such as albumin, cholesterol, manganese, iron, and copper. However, the relaxivities of these materials have not been determined yet. In this work, five sets containing the ions, albumin, and cholesterol were prepared by addition of increasing concentration of one material to each set. The relaxation times in these sets were measured by MRI, and the relaxation rates were fitted versus concentrations. The slopes of the fits were used as relaxivities. The (r 1, r 2) values of manganese, iron, and copper in mM−1 s−1, and those of albumin and cholesterol in (g/dl)−1 s−1 were found to be (32.64, 89.77), (0.31, 1.19), (0.5, 1.479), (0.01, 0.066) and (0.03, 0.458), respectively. The r 2/r 1 ratio ranged from 2.75 to 15.27. Manganese is an efficient relaxer, but iron and copper are poor ones. Albumin and cholesterol are efficient relaxers for only T 2. The contribution of water associated with native manganese of the cystic fluid to T 1 was 0.268 s−1, whereas those of water associated with native manganese, albumin, cholesterol, and iron to T 2 were 0.736, 0.185, 0.092, and 0.076 s−1, respectively. The other contributions were much smaller than 0.076 s−1. Manganese is most likely the compound altering T 1-weighted images between different jaw cysts, whereas manganese and albumin are most likely the compounds altering the T 2-weighted images. Present data suggest that such alterations may be used to separate jaw cysts from other jaw masses. The high r 2/r 1 suggests that T 2 is a more convenient parameter than T 1 for diagnostic use. This work is a part of the PhD thesis of U. Nezih Yilmaz supervised by R. Guner.  相似文献   

5.
We previously isolated an analog to chlorophyll-related compounds, pheophytin a, from the marine brown alga Sargassum fulvellum and demonstrated that it is a neurodifferentiation compound. In the current study, we investigated the effects of the pheophytin a analog vitamin B12 on PC12 cell differentiation. In the presence of a low level of nerve growth factor (10 ng ml−1), vitamin B12 demonstrated neurite outgrowth-promoting activity in PC12 cells. The effect was dose-dependent in the range of 6–100 μM. In the absence of nerve growth factor, vitamin B12 did not promote differentiation. To investigate the mechanism for this effect, we conducted differentiation assays and western blot analysis with signal transduction inhibitors and found that vitamin B12 did not promote PC12 cell differentiation in the presence of K252a or U0126 inhibitors. These results suggest that vitamin B12 stimulates PC12 cell differentiation through enhancement of the mitogen-activated protein kinase signal transduction pathway, which is also induced by nerve growth factor. Thus, vitamin B12 may be a good candidate for treatment of neurodegenerative diseases such as Alzheimer’s disease.  相似文献   

6.
One of the major effects of fluoride on oral bacteria is a reduction in acid tolerance, and presumably also in cariogenicity. The reduction appears to involve transport of protons across the cell membrane by the weak acid HF to dissipate the pH gradient, and also direct inhibition of the F1F0, proton-translocating ATPases of the organisms, especially for Streptococcus mutans. This direct inhibition by fluoride was found to be dependent on aluminum. The dependence on aluminum was indicated by the protection against fluoride inhibition afforded by the Al-chelator deferoxamine and by loss of protection after addition of umolar levels of Al3+, which were not inhibitory for the enzyme in the absence of fluoride. The F1 form of the enzyme dissociated from the cell membrane previously had been found to be resistant to fluoride in comparison with the F1F0 membrane-associated form. However, this difference appeared to depend on less aluminum in the F1 preparation in that the sensitivity of the F1 enzyme to fluoride could be increased by addition of umolar levels of Al3+. The effects of Al on fluoride inhibition were apparent when enzyme activity was assayed in terms of phosphate release from ATP or with an ATP-regenerating system containing phosphoenolpyruvate, pyruvate kinase, NADH and lactic dehydrogenase. Also, Be2+ but not other metal cations, e.g. Co2+, Fe2+, Fe3+, Mn2, Sn2+, and Zn2+, served to sensitize the enzyme to fluoride inhibition. The differences in sensitivities of enzymes isolated from various oral bacteria found previously appeared also to be related to differences in levels of Al. Even the fluoride-resistant enzyme of isolated membranes of Lactobacillus casei ATCC 4646 could be rendered fluoride-sensitive through addition of Al3+. Thus, the F1F0 ATPases of oral bacteria were similar to E1E2 ATPases of eukaryotes in being inhibited by Al-F complexes, and the inhibition presumably involved formation of ADP-Al-F inf3 sup- complexes during catalysis at the active sites of the enzymes.  相似文献   

7.
Mineral nutrition of poppy (Paparer somniferum L.) was studied in its effects on morphine production. Hydroponic cultures were carried out with nutritive solutions percolating over sand. The anion NO3- is the most efficient form of nitrogen for the production of fresh matter, dry matter or total morphine; in the latter respect, it rates higher than the NH4NO3 form which, on the other hand, gives higher alkaloid contents, while both cation NH4+ and urea have depressive effects. Phosphates have apparently little effect on the growth of the poppy, but solutions enriched with assimilable phosphate do stimulate flower proliferation and fruit development, without increasing markedly the total morphine output. Mg2+ and Ca2+ are important factors; Mg deficiencies will bring about a marked elongation of stems and early flowering, without any notable decrease in morphine outputs; conversely, Ca deficiencies will cause a drop in alkaloid production, while more calcium in the solution will give stronger elongation, a larger number of capsules and a marked increase in the weight of dry matter and morphine outputs, without any marked change in content. Sodium will favour poppy development (flowers and capsules) and will increase both the content and output of morphine. Na+ should therefore be introduced whenever possible in the fertilizing of poppy crops.  相似文献   

8.
ATP synthases, widely distributed in bacteria, eukaryotic mitochondria and chloroplasts, are highly conserved multi-subunit complexes. Although the conserved acidic residue in the transmembrane helix of the c subunit functions in H+ transport, the surrounding residues differ among species. Such divergence could lead to different regulatory modes since pH-dependent H+ transport has been demonstrated in E. coli with a c subunit carrying an additional acidic residue in the helix. There is further divergence in the number of c subunits that form the ring structure which is determined by the higher ordered structure. Recently, it was suggested that certain chemicals recognize the a and c subunits of pathogenic bacterial F0. Since there may be structural divergence even in well-conserved ATP synthases, the c subunit-ring as well as the a subunit in F0 could be targets for drugs for specific bacterial species.  相似文献   

9.
Abstract

The first complete mechanistic analysis of halide ion oxidation by a peroxidase was that of iodide oxidation by horseradish peroxidase. It was shown conclusively that a two-electron oxidation of iodide by compound I was occurring. This implied that oxygen atom transfer was occurring from compound I to iodide, forming hypoiodous acid, HOI. Searches were conducted for other two-electron oxidations. It was found that sulfite was oxidized by a two-electron mechanism. Nitrite and sulfoxides were not. If a competing substrate reduces some compound I to compound II by the usual one-electron route, then compound II will compete for available halide. Thus compound II oxidizes iodide to an iodine atom, I·, although at a slower rate than oxidation of I- by compound I. An early hint that mammalian peroxidases were designed for halide ion oxidation was obtained in the reaction of lactoperoxidase compound II with iodide. The reaction was accelerated by excess iodide, indicating a co-operative effect. Among the heme peroxidases, only chloroperoxidase (for example from Caldariomyces fumago) and mammalian myeloperoxidase are able to oxidize chloride ion. There is not yet a consensus as to whether the chlorinating agent produced in a peroxidase-catalyzed reaction is hypochlorous acid (HOCl), enzyme-bound hypochlorous acid (either Fe–HOCl or X–HOCl where X is an amino acid residue), or molecular chlorine Cl2. A study of the non-enzymatic iodination of tyrosine showed that the iodinating reagent was either HOI or I2. It was impossible to tell which species because of the equilibria:

I2+H2O=HOI+I-+H+</ p>

I-+I2=I3-

The same considerations apply to product analysis of an enzyme-catalyzed reaction. Detection of molecular chlorine Cl2 does not prove it is the chlorinating species. If Cl2 is in equilibrium with HOCl then one cannot tell which (if either) is the chlorinating reagent. Examples will be shown of evidence that peroxidase-bound hypochlorous acid is the chlorinating agent. Also a recent clarification of the mechanism of reaction of myeloperoxidase with hydrogen peroxide and chloride along with accurate determination of the elementary rate constants will be discussed.  相似文献   

10.
The present study was undertaken to investigate the effects of selenite (SeIV) and selenate (SeVI) on the all-trans retinoic acid (RA)-nuclear retinoic acid receptor (RAR) complex formation in rat liver. We also present the data on the in vitro effects of SeIV on the RARα and the type I iodothyronine 5′-deiodinase gene expression in the GH4C1 rat pituitary tumor cells. SeIV at 1.0 μmol/L was found to reduce (p<0.05) the RA specific binding to RAR in rat liver. Dithiothreitol (DTT), a protective agent for sulfhydryl groups, was found to be slightly effective in protecting the RAR binding properties when affected by SeIV. SeVI at 0.1 μmol/L reduced (p<0.05) the RA specific binding to RAR in liver, as well. Seleno-l-methionine (Se-II) when compared tol-methionine did not exert any inhibitory effect on the formation of the RA-RAR complex. SeIV (up to 2.5 μmol/L) has no inhibitory effect on GH4C1 cell proliferation as well as the prolactin secretion. SeIV at 1.0 μmol/L significantly decreases the rate of mRNA synthesis and/or degradation of the α form of the RAR and causes the enhancement of the type I iodothyronine 5′-deiodinase gene expression in GH4C1 cells. The results based on in vitro experiments suggest that inorganic selenium may affect the RA specific binding to their cognate receptor molecules, and it may reduce expression of the gene encoding the RARα, with the cell vitality and the cell growth remaining unchanged.  相似文献   

11.
Ion exchange properties of plant root cell walls   总被引:1,自引:0,他引:1  
Meychik  N.R.  Yermakov  I.P. 《Plant and Soil》2001,234(2):181-193
Acid-base properties and the swelling capacity of wheat, lupin and pea root cell walls were investigated. Roots of seedlings and green plants of different age were analysed by the potentiometric method. The ion exchange capacity (S i) and the swelling coefficient (K cw) of root cell walls were estimated at various pH values (from 2 to 12) and at different ionic strength (between 0.3 and 1000 mM). To analyse the polysigmoid titration curves pHi = f (S i), the Gregor's equation was employed. It was shown that the Gregor's model fits well the experimental data. The total number of the cation exchange (S t cat) and the anion exchange (S t an) groups were determined in the root cell walls. The number of the functional group of each type (S j) was estimated, and the corresponding values of pK a j were calculated. It was shown that for all types of cation exchangeable groups arranged in the cell wall structure the acid properties are enhanced by the increasing concentration of electrolyte. For each ionogenic group the coefficients of Helfferich's equation [pK a j = f (C K+)] were determined. It was found that the swelling of root cell walls changes with pH, C K+ and strongly depends on plant species. Within the experimental pH and C K+ range the swelling coefficient changes as follows: lupin > pea > wheat. The obtained results show that for the plant species under investigation the differences in the swelling coefficients originate from (a) the differences in the cross-linking degrees of polymeric chains arranged in the cell wall structure, (b) the differences in the number of carboxyl groups and (c) the differences in the total number of functional groups. Based on the estimated swelling coefficients in water it could be inferred that for wheat the cross-linking degree of the polymeric chains in the root cell walls is higher than those for lupin or pea. It has been emphasized that the calculated parameters (S j, pK a j, K cw), the equation {pK a j = f (CK+)} and the dependencies {K cw = f (CK+, pH)} allow to estimate quantitatively the changes in the ion exchange capacity of the root cell walls in response to the changes in an ionic composition of an outer solution. The results of these estimations allow to suggest that (a) the root apoplast is a compartment where the accumulation of cations takes place during the first stage of cation uptake from an outer medium, and (b) the accumulation degree is defined by pH and ionic composition of an outer solution. On the basis of the literature review and the results of the present experimental study it was proposed that the changes in the cell wall swelling in response to variances of environmental or experimental conditions could lead to a change of the water flow through a root apoplast. It has been supported that there is direct relationship between the swelling of root cell walls and the water flow within the plant root apoplast.  相似文献   

12.
Since the introduction of allozyme methods inthe mid 1960s it has been a standard practiceto report Wright's measure of populationsubdivision, F ST, for surveys ofgenetic variation. Its widespread use hasprovided us with a sense of what values can beexpected in particular situations and how theycan be interpreted. With some theoreticaljustification, F ST has also beenused to estimate rates of gene flow. Howeverthere are conditions under which F STis inappropriate for gene flow estimation andcan lead to incorrect or even absurdconclusions. These pitfalls have promptedcritics to suggest that F ST hasfailed to deliver what its proponents havepromised and should be abandoned. A furtherchallenge has been the development of newmethods that offer even greater promise. Thusit is reasonable to ask if perhaps it is timeto retire F ST and turn to new andmore powerful methods for the inference of geneflow from genetic markers. Here I will arguethat although gene flow should be estimated bymore powerful approaches whenever practical,F ST remains a useful measure of theaverage effects of gene flow and will continueto be used for comparative purposes.  相似文献   

13.
The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae) is one of the most important pests of cruciferous plants throughout the world. In recent years, this insect has been a serious pest for cabbage fields in Tehran province. Resistance of P. xylostella to all main groups of insecticides has been recorded and it is ranked in the 20 most resistant pest species reported up to now. According to many researchers, to eliminate the problem of pest resistance to chemical pesticides, an integrated pest management programme should be used. In line with this, the uses of microbial control agents (MCAs) are discussed. The bacterium, Bacillus thuringiensis (Bt) is one of microbial control agents of pests. It is characterised by its ability to produce proteic crystalline inclusions during sporulation. Cry1 protein has insecticidal activity and is highly specific to certain insects and not toxic to unrelated insects, plants or vertebrates. In this work, the pathogenicity of some Bt isolates, including Dipel, 20, 29, 79 and 87, was tested against P. xylostella and the lethal concentrations (LC50) of their crystal proteins to P. xylostella third larval instar was determined. The experiment was designed in factorial in randomised complete design with 5 treatments (different concentrations including 104, 105, 106, 107, 108 CFU/ml and 5 replications and with 10 third larval instars. Spore–crystal complex was applied to the surface of natural diets (cabbage leaves) and the mortality of P. xylostella larvae was assessed 120?h after exposure of Bt toxin in each treatment. Results showed that percentage of survival was significantly higher for control treatment. Results also showed that after 5?days, LC50 for isolates of Dipel, 20, 29, 79 and 87 were equal to 1?×?106, 1?×?105, 5?×?105, 4?×?105 and 1?×?104 CFU/ml, respectively. LT50 were equal to 93.71, 48.04, 71, 40.49 and 75.28?h. Of and most the percentage larval mortality relate to attendance 87 and also at least percentage mortality is related to the groom Dipel.  相似文献   

14.
KATP channels regulate insulin secretion by coupling β-cell metabolism to membrane excitability. These channels are comprised of a pore-forming Kir6.2 tetramer which is enveloped by four regulatory SUR1 subunits. ATP acts on Kir6.2 to stabilize the channel closed state while ADP (coordinated with Mg2+) activates channels via the SUR1 domains. Aberrations in nucleotide-binding or in coupling binding to gating can lead to hyperinsulinism or diabetes. Here, we report a case of diabetes in a 7-mo old child with compound heterozygous mutations in ABCC8 (SUR1[A30V] and SUR1[G296R]). In unison, these mutations lead to a gain of KATP channel function, which will attenuate the β-cell response to increased metabolism and will thereby decrease insulin secretion. 86Rb+ flux assays on COSm6 cells coexpressing the mutant subunits (to recapitulate the compound heterozygous state) show a 2-fold increase in basal rate of 86Rb+ efflux relative to WT channels. Experiments on excised inside-out patches also reveal a slight increase in activity, manifested as an enhancement in stimulation by MgADP in channels expressing the compound heterozygous mutations or homozygous G296R mutation. In addition, the IC50 for ATP inhibition of homomeric A30V channels was increased ~6-fold, and was increased ~3-fold for both heteromeric A30V+WT channels or compound heterozygous (A30V +G296R) channels. Thus, each mutation makes a mechanistically distinct contribution to the channel gain-of-function that results in neonatal diabetes, and which we predict may contribute to diabetes in related carrier individuals.  相似文献   

15.
A panel of 18 protein tyrosine kinase antagonists were tested for their inhibitory effect on human P2X7 receptor-mediated 86Rb+ (K+) efflux. The most potent compound (compound P), a phthalazinamine derivative and an inhibitor of vascular endothelial growth factor receptor kinase, blocked ATP-induced 86Rb+-efflux in human B-lymphocytes and erythrocytes by 76% and 66%, respectively. This inhibition was dose-dependent in both cell types with an IC50 of ∼5 μM. Kinetic analysis showed compound P was a non-competitive inhibitor of P2X7. This compound also inhibited ATP-induced ethidium+ influx into B-lymphocytes and P2X7-transfected-HEK-293 cells, as well as ATP-induced 86Rb+-efflux from canine erythrocytes. Externally, but not internally, applied compound P impaired ATP-induced inward currents in P2X7-transfected-HEK-293 cells. This study demonstrates that a novel protein tyrosine kinase antagonist directly impairs native and recombinant human P2X7 receptors. The data suggests that antagonists which target ATP-binding sites of kinases may potentially block the P2X7 receptor.  相似文献   

16.
We have made simultaneous spectrophotometric and O2 measurements on suspensions of rat neutrophils during activation of the respiratory burst. Under aerobic conditions an absorption increase attributable to myeloperoxidase compound II was observed in parallel with the rapid phase of O2 uptake. Identification of this compound was confirmed by analysis of a spectrum obtained with purified myeloperoxidase and H2O2. Whereas a second addition of stimulus did not increase O2 uptake any further, a second phase of myeloperoxidase release and compound II formation was observed. These results suggest thatin vivo myeloperoxidase reacts with H2O2 generatedvia the respiratory burst to form compound II under conditions in which the chlorination reaction would be the expected major pathway.Abbreviations FMLP N-formylmethionylleucyl phenylalanine - MPO2+.H2O2 Myeloperoxidase compound II - MPO3+.H2O2 Myeloperoxidase compound I - {ei275-1} superoxide  相似文献   

17.
Protobothrops flavoviridis venom contains plural phospholipase A2 (PLA2) isozymes. A [Lys49]PLA2 called BPII induced cell death in human leukemia cells. PLA2, an [Asp49]PLA2 that has much stronger lipolytic activity than BPII, failed to induce cell death. BPII-treated cells showed morphological changes, DNA fragmentation, and nuclear condensation. This BPII-induced apoptotic cell death was neither inhibited by inhibitors of caspases 3 and 6 nor accompanied by activation of procaspase 3, indicating that BPII-induced cell death is caspase independent. Since inactive p-bromophenacylated BPII induced cell death, BPII-induced apoptotic cell death is independent of PLA2 lipolytic activity. Rapid externalization of phosphatidylserine in BPII-treated cells was observed for fluorescein isothiocyanate (FITC)-labeled annexin V. In the cells treated with BPII, this spread over the cell membranes, implying that the cell toxicity of BPII is mediated via its cell-surface receptor.  相似文献   

18.
Ammonia often has been reported to inhibit cell growth. The aqueous ammonia equilibrium between the un-ionized form (NH3) and the ammonium ion (NH4 +) depends on the pH of the solution. Extensive studies in batch and continuous cultivation by varying pH and total ammonia concentration were carried out to investigate whether a kinetic model describing growth inhibition by ammonia has to be based on the total ammonia concentration, or the concentration of NH3. A significant relationship between the specific growth rate and death rate, respectively, and the NH3 concentration but not the total ammonia concentration, was detected. An adaptation of the cells to high ammonia levels was not observed. Based on these results a new kinetic model for ammonia mediated growth inhibition is suggested. For high density cultivation it is recommended to control the pH at the lower limit of the growth optimum to keep the NH3 level low.  相似文献   

19.
[目的] 为探究重金属对淡水绿藻生长的影响。[方法] 选取对水质检测具有明显指示作用的普通小球藻(Chlorella vulgaris)为实验材料,CdCl2·2H2O和CrCl3·7H2O提供重金属离子,探究不同浓度Cr3+和Cd2+在单一和复合胁迫下对藻细胞浓度、叶绿素a及相关抗氧化酶活性的影响。[结果] 随着Cr3+和Cd2+浓度不断增加,藻细胞浓度呈先增长后下降趋势;叶绿素a含量呈现先下降后升高再下降的现象,浓度为1 mg/L的单一和复合胁迫下有最大值,且毒性作用表现为Cr3+ < Cd2+ < Cr3++Cd2+;与藻细胞膜相关的丙二醛(MDA)和过氧化氢(H2O2)含量随着重金属离子浓度的增大而增长;重金属离子浓度低于10 mg/L时对藻细胞内抗氧化酶系统中的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)表现为促进作用,而大于10 mg/L时具有抑制作用。[结论] 结果表明在单一或复合重金属胁迫下,普通小球藻会充分调动与抗逆性相关的酶来维持自身的正常生长。  相似文献   

20.
Aluminum chloride (AlCl3), a neurotoxic compound, inhibited ATP diphosphohydrolase activity of synaptosomes obtained from cerebral cortex of adult rats. The metal ion significantly inhibited ATPase and ADPase activities of the enzyme at all concentrations tested in vitro (0.01, 0.05, 0.5, 5 and 10 mM) in the presence of 1.5 mM calcium. When tested in the absence of Ca2+, and with increasing amounts of Al3+, enzyme activity remained below basal levels, suggesting that the trivalent cation Al3+ is not a substitute for the divalent cation Ca2+ in ATP-Ca2+ and ADP-Ca2+ complexes. The Al3+ inhibition was competitive with respect to Ca2+. The enzyme inhibition was reversed by the addition of deferoxamine (DFO). NaF significantly inhibited ATP diphosphohydrolase activity, and this inhibition was reversed by the addition of Ca2+ to the medium. Such inhibition was not potentiated by AlF4, which is an inhibitor of cation-transport ATPases.  相似文献   

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