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71.
Glutamine and Aspartate Loading of Synaptosomes: A Reevaluation of Effects on Calcium-Dependent Excitatory Amino Acid Release 总被引:5,自引:3,他引:2
Guinea-pig cerebral cortical synaptosomes were preincubated for 60 min with 100 microM D-aspartate, L-aspartate, or L-glutamate. The total D- plus L-aspartate content of the synaptosomal fraction increased to 235%, 195%, or 164%, respectively, of the control. Despite this no increase was seen in the very low KCl evoked, Ca2+-dependent release of aspartate. Preincubation with the three amino acids changed the synaptosomal glutamate content to 78% (D-aspartate), 149% (L-aspartate), or 168% (L-glutamate) of control. However there was no statistically significant effect of these preincubations on the extent of Ca2+-dependent glutamate release. Thus the Ca2+-dependent release of aspartate and glutamate is not determined by the total synaptosomal content of these amino acids. The addition of 0.1-0.5 mM glutamine to the incubation caused a massive appearance of glutamate in the extrasynaptosomal medium. Analysis of specific activities showed that glutamine was hydrolysed directly by an extrasynaptosomal glutaminase, and that intrasynaptosomal glutamate was predominantly labelled by uptake of this glutaminase-derived glutamate. No increase was seen in the extent of Ca2+-dependent release of glutamate (by fluorimetry) either after preincubation with glutamine or in the continued presence of glutamine. Thus we are unable to confirm reports that glutamine expands the transmitter pool of glutamate. The extrasynaptosomal glutaminase activity in the synaptosomal preparation was inhibited by Ca2+ and activated by phosphate. Identical kinetics were obtained with "free" brain mitochondria, confirming the origin of the glutamine-derived glutamate. 相似文献
72.
73.
Validation of signature polarlipid fatty acid biomarkers for alkane-utilizing bacteria in soils and subsurface aquifer materials 总被引:3,自引:0,他引:3
David B. Ringelberg John D. Davis Glen A. Smith Susan M. Pfiffner Peter D. Nichols Janet S. Nickels J.Michael Henson John T. Wilson Marylynn Yates Donald H. Kampbell Harvey W. Read Thomas T. Stocksdale David C. White 《FEMS microbiology letters》1989,62(1):39-50
Abstract Extractable cell membrane-derived polarlipid ester-linked fatty acids (PLFA) obtained from aerated soils gassed with methane or propane and from methane- and propane-oxidizing bacteria isolated from the soils were analyzed by capillary gas chromatography/mass spectrometry. Exposure of aerated soils to methane resulted in the formation of a high proportion of an unusual 18-carbon mono-unsaturated PLFA, 18:lw8c. High proportions of this fatty acid biomarker are found in monocultures from this soil grown in minimal media with methane. This PLFA has been previously established as associated with authentic type II methane-oxidizing bacteria. The microbiota in aerated soils exposed to hydrocarbons containing propane, formed a suite of PLFA characterized by high proportions of a 16-carbon mono-unsaturated acid, 16:lw6c, and an 18-carbon saturated fatty acid with an additional methyl branch at the 10 position, 10 Me 18:0. This PLFA pattern has been detected in several monocultures enriched from the soil with propane-amended minimal media. The correspondence of high proportions of these unusual mono-unsaturated PLFA in the isolated monocultures and in situ in the soils after stimulation with the appropriate hydrocarbon is a strong validation of the utility of these biomarkers in defining the community structure of the surface soil microbial community. 相似文献
74.
The theory of interaction parameters has thus far been based on the free-energy relationships in the formation of ternary complexes formed between a pair of ligands and a protein molecule. The concept has been formulted in terms of a thermodynamic square comprised of the free protein, the two binary complexes, and the ternary complex. However, an increasing number of proteins have been found to exist as equilibrium mixtures of two macrostates. The equilibrium constants for such two-state transitions vary quite considerably between the various binary and ternary complexes of a given protein. We show here that the interpretations of interaction parameters in such two-state systems, requiring the use of a thermodynamic cube, are much more complex than those based on the classic thermodynamic square commonly employed. We demonstrate the use of enthalpies of interaction and heat capacities of interaction to analyze the source of observed free enerigies of interaction in such systems. Specifically, we find that measured negative interaction parameters may arise simply from the inability of a system to achieve all of the positive component effects anticipated by the conventional formulation. 相似文献
75.
Rapid detection of chromosome aneuploidies in uncultured amniocytes by using fluorescence in situ hybridization (FISH). 总被引:24,自引:4,他引:20 下载免费PDF全文
K Klinger G Landes D Shook R Harvey L Lopez P Locke T Lerner R Osathanondh B Leverone T Houseal 《American journal of human genetics》1992,51(1):55-65
Herein we report the results of the first major prospective study directly comparing aneuploidy detection by fluorescence in situ hybridization of interphase nuclei with the results obtained by cytogenetic analysis. We constructed probes derived from specific subregions of human chromosomes 21, 18, 13, X, and Y that give a single copy-like signal when used in conjunction with suppression hybridization. A total of 526 independent amniotic fluid samples were analyzed in a blind fashion. All five probes were analyzed on 117 samples, while subsets of these five probes were used on the remaining samples (because of insufficient sample size), for a total of over 900 autosomal hybridization reactions and over 400 sex chromosome hybridization reactions. In this blind series, 21 of 21 abnormal samples were correctly identified. The remaining samples were correctly classified as disomic for these five chromosomes. The combination of chromosome-specific probe sets composed primarily of cosmid contigs and optimized hybridization/detection allowed accurate chromosome enumeration in uncultured human amniotic fluid cells, consistent with the results obtained by traditional cytogenetic analysis. 相似文献
76.
Charlotte L. Branchaud Cynthia G. Goodyer Harvey J. Guyda Yves Lefebvre 《In vitro cellular & developmental biology. Plant》1990,26(9):865-870
Summary We have compared hormone production by early gestation and term human placental trophoblasts cultured in Ham's F10 medium
containing 10% fetal bovine serum with that by cells cultured in serum-free HB102 medium. Mean daily production of progesterone
on Days 3 to 7 was approximately 25% less by both early gestation and term cells cultured in HB102 as compared to Ham's F10,
but production was maintained at a stable level for at least 7 d longer than the cells in Ham's. Estradiol production from
10−6
M dehydroepiandrosterone by both early gestation and term cells was comparable in both media. Human placental lactogen production
on Days 3 to 7 was 40% less by cells cultured in HB102. Human chorionic gonadotropin (hCG) output by early gestation cells
was also 50% less in HB102 but term cells in HB102 produced twice as much hCG as those in Ham's F10. 3B-Hydroxysteroid dehydrogenase
(3BHSD) activity in early gestation and term cells and 11B-hydroxysteroid dehydrogenase (11BHSD) activity of early gestation
cultures was comparable in the two media. 11BHSD activity was decreased in the term cultures, and this decrease was more marked
in Ham's than in HB102. Sulfatase and aromatase activities in the early gestation cultures were comparable in both media;
sulfatase activity was comparable and aromatase activity only 20% less in the term cells cultured in HB102. These results
indicate that serum-free HB102 supports differential function of human trophoblast cells and is useful for studies of placental
activity for as long as 14 d in culture. 相似文献
77.
Continuous pollen monitoring of an urban network consisting of three stations has been undertaken for a period of 2 years
in Perugia, central Italy. The aim has been to establish whether the Perugia pollen trap, active since 1983, is still representative
of the area following recent urbanisation. Quantitative differences were found between the stations, reflecting different
vegetational areas, but only slight differences were detected in relation to the timing of the principal period of pollination.
Therefore, although individual pollen traps are necessary to characterize fully the different areas, one trap is sufficient
to determine the key allergenic thresholds in the studied area. 相似文献
78.
Continuous pollen monitoring of an urban network consisting of three stations has been undertaken for a period of 2 years in Perugia, central Italy. The aim has been to establish whether the Perugia pollen trap, active since 1983, is still representative of the area following recent urbanisation. Quantitative differences were found between the stations, reflecting different vegetational areas, but only slight differences were detected in relation to the timing of the principal period of pollination. Therefore, although individual pollen traps are necessary to characterize fully the different areas, one trap is sufficient to determine the key allergenic thresholds in the studied area. 相似文献
79.
In comparison to wild type Arabidopsis thaliana, the auxin resistant mutants axr1 and axr2 exhibit reduced inhibition of root elongation in response to auxins. Several auxin-regulated physiological processes are also altered in the mutant plants. When wild-type, axr1 and axr2 seedlings were grown in darkness on media containing indoleacetic acid (IAA), promotion of root growth was observed at low concentrations of IAA (10?11 to 10?7M) in 5-day-old axr2 seedlings, but not in axr1 or wild-type seedlings. In axr1 there was little or no measurable root growth response over the same concentration range. In wild type, root growth was inhibited at concentrations greater than 10?10M and no detectable root growth response was observed at lower concentrations. In addition, production of lateral roots in response to IAA increased in axr2 seedlings and decreased in axr1 seedlings relative to wild type. Promotion of root elongation and initiation of lateral roots in axr2 seedlings in response to auxin indicate that axr2 seedlings are able to perceive and respond to IAA. 相似文献
80.
Kirsty Barr Henrik Moller Emma Christmas Philip Lyver Jacqueline Beggs 《Oecologia》1996,105(2):266-270
An introduced social wasp Vespula vulgaris may compete with native birds for honeydew and invertebrates in New Zealand forests. Experimentally hidden mealworms (Tenebrio molitor) persisted longer at two sites following wasp poisoning that at two sites where wasps were not poisoned. Mealworms persisted longer in the morning than in the afternoon within all study sites. An unusually low mealworm removal rate during a morning trial before wasp poisoning heavily influences the results of this experiment but we have no ecological reason to ignore it. Wasps may therefore be having a heavy impact on invertebrate abundance on very short time scales (within a day following dawn emergence). They may also remove cached food items that would otherwise be retrieved by the South Island robin (Petroica australis australis) during cold or dark feeding conditions. 相似文献