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1.
Cynthia L. Deitrick Richard E. Katholi David J. Huddleston Kathy Hardiek Lucienne Burrus 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2001,751(2)
Vitamin B6, measured as pyridoxal 5′-phosphate (PLP), is a co-enzyme in the transsulfuration pathway of homocysteine metabolism. Since depletion of PLP has been suggested as an independent risk factor for coronary artery disease, PLP is frequently measured to guide patient care. By a change and utilization of an Aquasil C18 column and the addition of an acetonitrile clean-up gradient to the potassium phosphate, with sodium perchlorate and bisulfite buffer between samples we report the modification of a previously described method for analysis of PLP. The result is a more practical, efficient, reliable and robust method for daily clinical use. We also determined and report that it is critical to protect freshly prepared standard PLP samples from light exposure during assay preparation. 相似文献
2.
Intestinal cholesterol absorption is specifically inhibited by the 2-azetidinone cholesterol absorption inhibitor ezetimibe. Photoreactive ezetimibe analogues specifically label a 145-kDa protein in the brush border membrane of enterocytes from rabbit small intestine identified as aminopeptidase N (CD13). In zebrafish and mouse small intestinal cytosol, a heterocomplex of Mr 52 kDa between annexin II and caveolin 1 was suggested as a target of ezetimibe. In contrast, in the cytosol and brush border membrane vesicles (BBMV) from rabbit small intestine of control animals or rabbits treated with the nonabsorbable cholesterol absorption inhibitor AVE 5530, both annexin II and caveolin 1 were exclusively present as monomers without any heterocomplex formation. Upon immunoprecipitation with annexin II a 52-kDa band was observed after immunostaining with annexin II antibodies, whereas no staining of a 52-kDa band occurred with anti-caveolin 1 antibodies. Vice versa, a 52-kDa band obtained by immunoprecipitation with caveolin 1 antibodies did not stain with annexin II-antibodies. The intensity of the 52-kDa band was dependent on the amount of antibody and was also observed with anti-actin or anti-APN antibodies suggesting that the 52-kDa band is a biochemical artefact. After incubation of cytosol or BBMV with radioactively labelled ezetimibe analogues, no significant amounts of the ezetimibe analogues could be detected in the immunoprecipitate with caveolin-1 or annexin II antibodies. Photoaffinity labelling of rabbit small intestinal BBMV with ezetimibe analogues did not result in labelling of proteins being immunoreactive with annexin II, caveolin 1 or a 52-kDa heterocomplex. These findings indicate that the rabbit small intestine does not contain an annexin II/caveolin 1 heterocomplex as a target for ezetimibe. 相似文献
3.
4.
B. C. Loughman 《Plant and Soil》1987,99(1):63-74
Summary Little is known about the biochemical basis of the genotypic differences in the capacity for ion absorption and transport
shown by many crop species. If these differences reflect the abundance of a specific membrane component or the activity of
an enzyme we need to have some indication of thein vivo operation of these systems in whole plants. Thein vivo assessment of glycolytic enzymes is illustrated by the effects of mannose on the transport of phosphate in maize varieties.
The application of high resolution31P-NMR to the study of intermediary metabolismin vivo is also helpful in following transport capacity.
The five-fold rise in respiratory rate that occurs when freshly cut potato slices are maintained in aerated water for 24 hours
is accompained by the turning on of a wide range of biochemical systems. Major increases in the capacity for absorption of
phosphate from low concentrations (0.1 μM–10 μM) and in the phosphorylative ability of the tissue are seen, indicating the synthesis of a carrier involved in phosphate transport.
These capacities differ markedly between individual tissues of the tuber,i. e. pith, parenchyma, cortex and buds and large differences have been observed between comparable tissue from different varieties.
Varieties grown under similar conditions have been compared and shown to exhibit different kinetics with respect to the development
of the low concentration absorption site and in their sensitivity to the effects of uncouplers such as 2,4-dinitrophenol. 相似文献
5.
C. F. C. Bonting H. W. van Veen A. Taverne G. J. J. Kortstee A. J. B. Zehnder 《Archives of microbiology》1992,158(2):139-144
The response of Acinetobacter strain 210A to low phosphate concentrations was investigated in P- or C-limited chemostat cultures. The organism accumulated poly--hydroxybutyric acid under P-deprivation, at phosphate concentrations ranging from 0.1 to 0.7 mM. The amount of biomass was proportional to the phosphate concentration in the medium and no polyphosphate was formed. When shifting a culture from P- to C-limitation phosphate was accumulated as polyphosphate. No poly--hydroxybutyrate could be detected in these cells. The amount of polyphosphate in the cell showed a hysteresis. When cultures were shifted from low to high phosphate concentrations, polyphosphate reached a maximum of about 60 mg P per gram of dry weight at about 3 times excess phosphate (ca. 2.5 mM Pi). It decreased to 45 mg P per gram dry weight at approximately 5 times the phosphate needed for growth (ca. 3.5 mM Pi). In the reverse case (high to low) polyphosphate did never exceed 45 mg P per gram dry weight. The specific activities of alkaline phosphatase and the phosphate uptake system were induced at residual Pi concentrations below the detection limit (<10 M). The specific uptake rate followed also a hysteresis. The specific activities of polyphosphatase and polyphosphate: AMP phosphotransferase increased when polyphosphate formation was possible.Abbreviations HPP
High polymeric polyphosphates
- PHB
Poly--hydroxybutyric acid
- PPn
Polyphosphate
- PQQ
Pyrrolo-quinoline quinone
- U
1 mol product formed · min-1 相似文献
6.
Summary Phosphate sorption isotherms were developed for five Philippine wetland rice soils using the conventional technique and a modified one. In the conventional method, P requirements of soils varied between 280 and 810 g P/g soil. In the modified method, they varied from 160 to 540 g P/g soil at 0.2 ppm P in solution. Soils with high P-sorption capacities had vermiculite and halloysite as the dominant clay minerals. Soil reduction by flooding decreased P-sorption by 28–70 percent at 0.2 ppm P in solution. The decrease in P-sorption due to soil reduction was greatest in a crystalline soil with vermiculite and halloysite as the dominant clay minerals and least in a soil with dominant X-ray amorphous silicates in the clay fraction.Desorption of freshly adsorbed P under reduction was greater in HCO
3
–
solution than in CaCl2 and it increased with level of applied P. Desorption patterns of freshly adsorbed P were similar to adsorption patterns but values of P in solution were lower at desorption. Soils varied with respect to desorption of freshly sorbed P. Desorption studies indicate that soils vary in intensity factor with respect to P and thus influence P availability to plants. Use of P-sorption and P-desorption data obtained under reduced soil condition was proposed for detecting P needs of submerged rice soils.Results of a pot study with IR36 at different levels of solution P (reduced) in one soil indicated a high degree of correlation between adjusted P levels and the measured growth parameters. About 0.12 ppm P in the soil solution or 0.46 ppm P desorbed in HCO
3
–
solution (equivalent to 100 mg P/kg soil) was adequate for near-maximum plant height, tiller production, total dry matter yield, plant P content, and total P uptake. 相似文献
7.
The effect of phosphate feeding on the influence of low (2%) oxygen on photosynthetic carbon assimilation has been investigated in leaf discs of spinach (Spinacia oleracea L.) at 12°C. The following observations were made. First, after the transition from 20% O2 to 2% O2, the rate of CO2 uptake was inhibited at CO2 concentrations between about 250 and about 800 l CO2·l-1. Second, phosphate feeding stimulated the rate of CO2 uptake in 20% O2 at higher concentrations of CO2 (500–900 l·l-1). Third, phosphate feeding stimulated the rate of CO2 uptake in 2% O2 at all but the highest (900 l·l-1) and lowest 74 (l·l-1) concentrations of CO2 employed. Phosphate thereby restored the stimulation of photosynthesis by 2% O2 and it did so over a wide range of lower temperatures. Fourth, oscillatory behaviour, however generated, was dampened by phosphate feeding, even at very low concentrations of CO2. Contents of leaf metabolites were measured during the transition to 2% O2 in control and phosphate-fed leaf discs. During this period the ratio glycerate-3-phosphate/triose phosphate rose steeply, but fell again only in the phosphate-treated leaf discs. These data, taken together with measured ATP/ADP ratios, showed that assimilatory power, the ratio [ATP]·[NAD(P)H]/[ADP]·[Pi]·[NAD(P)], decreased when leaves were exposed to 2% O2, but that this decrease was minimised by previous feeding of phosphate. The mechanism of phosphate limitation is discussed in the light of the results.Abbreviations Ci
intercellular concentration of CO2
- RuBP
ribulose-1,5-bisphosphate 相似文献
8.
Metabolic acidosis produces a phosphaturia which is independent of parathyroid hormone or dietary phosphorus intake. To study the underlying mechanism, inorganic phosphate (Pi) and glucose transport were studied in brush-border membrane vesicles prepared from the renal cortex of parathyroidectomized rats gavaged for three days with either 7.5 ml of 1.6% NaCl (control) or 1.5% NH4Cl (acidosis). At killing, blood pH and plasma bicarbonate were and , respectively, in control and () and () in acidotic rats. Serum Pi was similar in both groups, while 24 h urine Pi excretion was higher in the acidotic group (). Peak sodium-dependent uptake of Pi, measured after 1.5 min of incubation, was higher in controls than acidotic rats ( vs. protein, ), whereas peak glucose uptake at 1.5 min was not significantly different between the groups. Equilibrium values for Pi and glucose uptake were similar in the two groups. for Pi uptake in the control and acidotic animals were not different, 0.036 and 0.040 mM, respectively. By contrast, was higher in controls than in the acidotic group, 3.13 vs. 1.15 nmol/mg protein per 15 s. These results suggest that metabolic acidosis directly inhibits Pi uptake by the brush border of the proximal tubule by decreasing the availability of Pi carriers of the renal brush-border membrane. 相似文献
9.
The techniques of 27Al- and 31P-nuclear magnetic resonance (NMR) spectroscopy were used to investigate the interactions of aluminium with intracellular ligands within the mycelium of the ectomycorrhizal basidiomycete Laccaria bicolor (Maire) Orton (S238). The vegetative mycelium was grown on medium containing 0.5 mM AlCl3 for 0.5 to 3 d. The 27Al-NMR spectra showed that aluminium was rapidly taken up and accumulated into polyphosphate complexes in the vacuole. Comparison with Al-polyphosphate complexes obtained in vitro on model systems indicated that Al forms at least three mixed-solvation complexes with Pi and polyphosphates, that there is more than one complex present under any set of conditions, and that the equilibrium between these complexes shifts dramatically with Al concentration in the medium. The high phosphate concentrations in the growth medium favoured the accumulation of the Al-polyphosphate complexes. When mycelium containing Al-polyphosphate complexes was transferred to Al-free nutrient solution for 9 d, the Alpolyphosphate complexes were not remobilized. The sequestration of Al in the polyphosphate complexes could therefore make a significant contribution to the protection of mycorrhizal plants against aluminium toxicity.Abbreviations NMR
nuclear magnetic resonance
- PolyP
polyphosphate(s)
- PP1
terminal phosphate of PolyP
- PP3
middle phosphate of PolyP
We thank Prof. Daniel Canet (Laboratoire de Méthodologie RMN, University of Nancy I, Vandceuvre-lès-Nancy, France) for his constant encouragement and Christine Delaruelle for skilled technical assistance in growing the fungal cultures. This work was supported by a research grant from the Commission of the European Communities (STEP-CT90-0059, Role of Ectomycorrhiza in Stress Tolerance of Forest Trees) to F.M. and a travel grant from the Institut National de la Recherche Agronomique to I.K.; R.C. is a recipient of a Postdoctoral Fellowship from the Natural Sciences and Engineering Research Council of Canada. 相似文献
10.
Summary Despite extensive discussions during the last 20 years stromatolites are still used by many geologists as unequivocal indicators
of very shallow-water conditions. We investigated four stratigraphic units from the Lower and Middle Jurassic of southern
Germany (Posidonien-Schiefer, Amaltheen-Ton) and of the Northern Calcareous Alps (Adneter Kalk, Klauskalk), which were formerly
interpreted as shallow marine sediments by some authors due to the occurrence of stromatolites. Our interpretations of the
macro-, micro- and ultrafacies of these sediments are not compatible with shallow-water settings. We therefore propose a deep-marine,
aphotic origin of these stromatolites.
Former interpretations of the Posidonien-Schiefer as a shallow-water deposit are mainly based on the occurrence of stromatolites.
We favour the model of a temporarily stagnant, deep, aphotic basin for these planktonrich sediments. Particles resembling
ooids, but lying within mudstones cannot be taken as evidence for shallow agitated water. They either formed within the mud
or are allochthonous.
The deep-water setting of the red limestone of the Alpine Early and Middle Jurassic is indicated by a lack of platform-typical
components like coated grains and phototrophic benthos and by shells of plankton and nekton forming a major part of the sediment.
Stromatolites occur on the steep slope of a drowned Rhaetian reef with an estimated relief of 50–100 m and immediately below
and within radiolarian limestones, deposited below the aragonite compensation depth (ACD).
The aphotic stromatolites show some morphological differences to their shallow water counterparts. In all of our sections
they occurred during intervals of reduced sedimentation. They form only thin horizons and probably grew very slowly. Mineralizations
by Fe−Mn oxides and phosphate are very common. The presence of a microbial film is evident from binding of sedimentary particles,
but the nature of the microbes is not known. Growth habits within the very distinct environments of red limestone and black
shales show some common features, but also clear differences.
The microproblematicumFrutexites
Maslov is a very common component in deep-water stromatolites, but may also itself form small crusts or dendrolites. It occurs in
two different forms. Opaque, slender forms with indistinct outlines probably grew within the weakly lithified sediment. Thicker,
transparent forms with well defined outlines are found in cavities and probably also grew on the seafloor. Well preserved
specimens display an internal fabric of radially arranged fibres of Fe−Mn oxides and calcite. It is suggested that calcite
or aragonite were one original mineralogy ofFrutexites, which was later replaced by Fe−Mn oxides or phosphate.
It is not certain whetherFrutexites is an organic, biomineralized structure or an inorganic mineralization, but the variable mineralogy and growth forms in different
environments point to an organic origin. But even if organic, the occurrence in cryptic habitats and negative phototactic
growth-directions make it clear thatFrutexites was not phototrophic. 相似文献