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61.
Marc Yudkoff Yevgeny Daikhin Zhi-Ping Lin Liana Nissim Janet Stern David Pleasure Itzhak Nissim 《Journal of neurochemistry》1994,62(3):1192-1202
Abstract: The aim was to study the extent to which leu-cine furnishes α-NH2 groups for glutamate synthesis via branched-chain amino acid aminotransferase. The transfer of N from leucine to glutamate was determined by incubating astrocytes in a medium containing [15N]leucine and 15 unlabeled amino acids; isotopic abundance was measured with gas chromatography-mass spectrometry. The ratio of labeling in both [15N]glutamate/[15N]leucine and [2-15N]glutamine/[15N]leucine suggested that at least one-fifth of all glutamate N had been derived from leucine nitrogen. At the same time, enrichment in [15N]leucine declined, reflecting dilution of the 16N label by the unlabeled amino acids that were in the medium. Isotopic abundance in [16N]-isoleucine increased very quickly, suggesting the rapidity of transamination between these amino acids. The appearance of 15N in valine was more gradual. Measurement of branched-chain amino acid transaminase showed that the reaction from leucine to glutamate was approximately six times more active than from glutamate to leucine (8.72 vs. 1.46 nmol/min/mg of protein). However, when the medium was supplemented with α-ketoisocaproate (1 mM), the ketoacid of leucine, the reaction readily ran in the “reverse” direction and intraastrocytic [glutamate] was reduced by ~50% in only 5 min. Extracellular concentrations of α-ketoisocaproate as low as 0.05 mM significantly lowered intracellular [glutamate]. The relative efficiency of branched-chain amino acid transamination was studied by incubating astrocytes with 15 unlabeled amino acids (0.1 mM each) and [15N]glutamate. After 45 min, the most highly labeled amino acid was [15N]alanine, which was closely followed by [15N]leucine and [15N]isoleucine. Relatively little 15N was detected in any other amino acids, except for [15N]serine. The transamination of leucine was ~17 times greater than the rate of [1-14C]leucine oxidation. These data indicate that leucine is a major source of glutamate nitrogen. Conversely, reamination of a-ketoisocaproate, the ketoacid of leucine, affords a mechanism for the temporary “buffering” of intracellular glutamate. 相似文献
62.
Abstract: Changes of the main hydrodynamic and oxygen transfer parameters during Aspergillus niger cultivation in an external-loop air-lift bioreactor of 200 dm3 operating capacity were investigated. The final average concentrations of biomass and citric acid obtained in batch fermentations were about 17 g 1-1 and 90 g 1-1 , respectively. Significant influence of the increasing biomass concentration on the rheological properties of the broth and operating parameters was found. Volumetric oxygen transfer coefficient. k L a , was found to be dependent on the apparent viscosity of the broth with an exponent of -0.984. 相似文献
63.
Presence and role of jasmonate in apple embryos 总被引:3,自引:0,他引:3
Rajiv Ranjan Otto Miersch Giinther Sembdner Stanistaw Lewak 《Physiologia plantarum》1994,90(3):548-552
(-)Jasmonic acid (JA) was identified in extracts front embryos of apple (Mulus domestica) by combined gas chromatography-mass spectromelry. Quantification of JA in embryos isolated from seeds at different perexts of stratification by gas chromalography combined with mass spectrometry/selective ion monitoring indicated a sharp peak at day 30. At the same time the maximal ratio of conjugated to free JA was found by enzyme-linked imrnunosorbent assay (ELISA). Germination of embryo.s was stimulated by added JA and inhibited by salieylhydroxamic acid (SHAM, an inhibitor of lipoKygenase). Both stimulation and inhibition disappeared in embryos stratified for more than 30 days. Methyl jasmonate was more effective in stimulation of embryo germination than free JA. while JA-isoleucine inhibited germination. The possible mechanism responsible for changes in JA level as wel! as the role of JA and its conjugates in removal of dormancy in apple seeds are discussed. 相似文献
64.
Surfactant effects on protein structure examined by electrospray ionization mass spectrometry. 总被引:1,自引:0,他引:1 下载免费PDF全文
R. R. Loo N. Dales P. C. Andrews 《Protein science : a publication of the Protein Society》1994,3(11):1975-1983
Electrospray ionization mass spectrometry (ESI-MS) has proven to be a useful tool for examining noncovalent complexes between proteins and a variety of ligands. It has also been used to distinguish between denatured and refolded forms of proteins. Surfactants are frequently employed to enhance solubilization or to modify the tertiary or quaternary structure of proteins, but are usually considered incompatible with mass spectrometry. A broad range of ionic, nonionic, and zwitterionic surfactants was examined to characterize their effects on ESI-MS and on protein structure under ESI-MS conditions. Solution conditions studied include 4% acetic acid/50% acetonitrile/46% H2O and 100% aqueous. Of the surfactants examined, the nonionic saccharides, such as n-dodecyl-beta-D-glucopyranoside, at 0.1% to 0.01% (w/v) concentrations, performed best, with limited interference from chemical background and adduct formation. Under the experimental conditions used, ESI-MS performance in the presence of surfactants was found to be unrelated to critical micelle concentration. It is demonstrated that surfactants can affect both the tertiary and quaternary structures of proteins under conditions used for ESI-MS. However, several of the surfactants caused significant shifts in the charge-state distributions, which appeared to be independent of conformational effects. These observations suggest that surfactants, used in conjunction with ESI-MS, can be useful for protein structure studies, if care is used in the interpretation of the results. 相似文献
65.
The inorganic contents of bone, brain, erythrocyte, heart, kidney cortex, kidney medulla, liver, lung, muscle and plasma from spontaneously hypertensive rats were compared with those of the same tissues from healthy Sprague-Dawley rats. A general inductively coupled plasma-mass spectrometry method developed for multi-element determinations of most of the elements present in biological tissues was used. Variations were found not only for major elements, as expected, but also for many trace elements in several tissues. 相似文献
66.
Daniel R. Mayer Walter Kosmus Helmut Pogglitsch David mayer Wolfgang Beyer 《Biological trace element research》1993,37(1):27-38
Serum arsenic concentrations of persons suffering from renal failure and undergoing hemodialysis treatment (n=85) and of healthy controls (n=25) were determined by hydride-generation AAS technique after microwave digestion. The results were evaluated by comparing
the values of both groups, considering physiological factors and individual data, as well as comorbid conditions of the hemodialysis
(HD) patients. Serum arsenic levels were diminished in the patient group compared with controls (mean values 8.5±1.8 ng/mL
vs 10.6±1.3 ng/mL). Furthermore, additional diseases within the hemodialysis group, particularly injuries of the central nervous
system (CNS), vascular diseases, and cancer, were correlated to occasionally markedly decreased serum arsenic concentrations.
It was concluded that arsenic homeostasis is disturbed by HD treatment and certain additional diseases. Desirable arsenic
concentrations in the body seem to be reasonable. This consideration results in the conclusion that arsenic could play an
essential role in human health. Thus, reference arsenic concentrations in different human tissues and body fluids should be
established in order to recognize not only arsenic intoxication, but also arsenic deficiency. Perhaps arsenic deficiency contributes
to the increased death risk of HD patients, and therefore, arsenic supplementations for patients with extremely low serum
arsenic concentrations should be taken into account. 相似文献
67.
Cobalt determinations in biological fluids are of great interest in biological or toxicological research programs. Cobalturia
is often chosen as an indicator for a biological monitoring program in occupational exposure to cobalt dusts. The method described
here derives from the IUPAC reference method for nickel determination. It enables cobaltemia and cobalturia to be measured
in small samples (1 mL). The mean usual values for cobalt in biological fluids are very low (2.7 nmol L−1 for serum and 6.7 nmol L−1 for urine), and therefore, thus require an analytical procedure with preconcentration and extraction. The sample is mineralized
by wet acid digestion. After digestion, inorganic cobalt is extracted in form of ammonium pyrrolidine dithiocarbamate complex
into isobutyl methyl ketone and measured in the organic layer by electrothermal atomic absorption spectrometry.
The analytical parameters are described in detail. The extraction output is about 99%. The detection limits are 1.93 and 1.89
nmol L−1 for serum and urine, respectively. Sensitivity (expressed as the concentration that gives a 0.044 absorbance) is 3.4 nmol
L−1 for serum and 3.3 nmol L−1 for urine.
Within-run precision ranged between 3.9 and 2.5% (coefficients of variation) for serum and 4.2 and 1.1% for urine, at 87 and
136 nmol L−1 levels, respectively. Between-run precision ranged between 4.3 and 3.3% (coefficients of variation) for serum and 4.2 and
2.3% for urine, at 87 and 136 nmol L−1 levels, respectively. At very low concentration, 5.7 nmol L−1 for serum and 2.5 nmol L−1 for urine, the between-run precision is, respectively, 19.5 and 28%.
Linearity is effective between 0 and 272 nmol L−1. Interferences and matrix effects are negligible for urine, serum, or plasma samples without hemoglobin. The method is easily
applicable for routine determinations. 相似文献
68.
Evidence for the Presence of "Metabolic Sterols" in Pneumocystis: Identification and Initial Characterization of Pneumocystis carinii Sterols 总被引:1,自引:0,他引:1
EDNA S. KANESHIRO JAYNE E. ELLIS KOKA JAYASIMHULU DAVID H. BEACH 《The Journal of eukaryotic microbiology》1994,41(1):78-85
Mixed life cycle stages of rat-derived Pneumocystis carinii were isolated from host lungs and their sterols were compared with those present in lungs from normal and immunosuppressed uninfected rats. Gas-liquid chromatography consistently detected, resolved, and quantified 9, 10, and 20 sterol components in the total nonsaponifiable neutral lipid fraction of lungs from normal rats, lungs from immunosuppressed uninfected rats, and P. carinii preparations, respectively. In all samples, cholesterol was the most abundant sterol present, comprising 97%, 93%, and 78% of total sterols in lungs from normal rats, lungs from immunosuppressed uninfected rats, and P. carinii , respectively. Tentative identifications of several rat lung and P. carinii minor sterols were made based on gas-liquid chromatogram retention times and fragmentation patterns from mass spectral analyses. Campesterol (ergost-5-en-3-ol), cholest-5-en-3-one, and β -sitosterol (stigmast-5-en-3-ol) were among the minor components present in both types of lung controls, and were also components of P. carinii sterols. In contrast to lung controls, the sterols of P. carinii were enriched in C28 and C29 sterols with one or two double bonds, and a hydroxyl group at C-3 (ergost-5-en-3-ol, ergost-7-en-3-ol, ergosta-dien-3-ol, stigmast-5-en-3-ol, stigmast-7-en-3-ol and stigmasta-dien-3-ol). Steryl esters of P. carinii , probably stored in cytoplasmic lipid droplets, were dominated by those present in the host lung. In separate studies. 3-hydroxy-3-methylglutaryl coenzyme A activity, a key enzyme in the regulation of sterol biosynthesis, was detected in purified P. carinii preparations and incorporation of radiolabeled squalene and mevalonate was observed. Together, these results suggest that the parasite readily takes up and incorporates host sterols, and that the organism synthesizes some of its own "metabolic sterols" 相似文献
69.
High-temperature gas chromatography and gas chromatography-inass spectrometry for the analyses of oligosaccharides derived
from glycoproteins or glycosphingolipids has been developed. Pcrmethylatcd oligosaccharides with up to about 12 sugar residues
and masses up to 2500 Daltons can be analyzed. This approach is discussed and exemplified. 相似文献
70.
William P. Jacobs 《Journal of phycology》1993,29(5):595-600
The best available methods were used to search for an-giosperm hormones in the green alga Caulerpa paspa-loides (Bory) Greville. Solvent partitioning of methanol extracts led to acidic ethyl acetate fractions that it were run through high-performance liquid chromatography. Each of the resulting 25 fractions was divided in half so that one half was tested in the dwarf rice or lettuce hypocotyl assays for gibberellins and the other half was methylated and silylated for capillary gas chromatography combined with computerized mass spectrometry. By comparing algal mass spectra with spectra in the extensive library of mass spectra and associated Kovats Retention Indices that MacMillan's Bristol group has amassed, indole-3-acetic acid's presence in the alga was confirmed and dioxindole-3-acetic acid was found. However, despite the presence of several peaks of gibberellin-like activity in the bioassays, no gibberellin or gibberellin metabolite was found in the thousands of full mass spectra examined. The gibberellin-like activity in the acidic ethyl acetate fractions was therefore presumably not from any gibberellin known from vascular plants. 相似文献