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171.
Kam H. Leung 《Cancer immunology, immunotherapy : CII》1991,34(1):31-36
Summary We have shown that depletion of monocytes from human peripheral blood mononuclear cells (PBMC) byl-phenylalanine methyl ester (PheOMe) enhanced lymphokine-activated killer cell (LAK) generation by recombinant interleukin-2 (rIL-2) at high cell density. In this study, we have investigated the mechanism of action of PheOMe on LAK activation by using trypsin, chymotrypsin, tosylphenylalaninechloromethanol (TPCK, a chymotrypsin inhibitor), tosyl-l-lysinechloromethane (TLCK, a trypsin inhibitor), phenylalaninol (PheOH), and benzamidine. PBMC were treated with 1–5 mM PheOMe for 40 min at room temperature in combination with the various agents, washed and assessed for their effects on natural killer (NK) activity against K562 cells and monocyte depletion. The treated cells were then cultured with or without rIL-2 for 3 days. LAK cytotoxicity was assayed against51Cr-labeled K562 and Raji tumor target cells. TPCK at 10 µg/ml partially inhibited depletion of monocytes by PheOMe. TLCK did not prevent depletion of monocytes nor inhibition of NK activity induced by PheOMe. TPCK and TLCK inhibited NK activity by themselves. TPCK but not TLCK inhibited rIL-2 induction of LAK cells. On the other hand, PheOH and benzamidine (analogs of PheOMe) lacked any effect on monocyte depletion but abrogated the inhibitory effect of PheOMe on NK activity. They had no effect on rIL-2 activation of LAK activity enhanced by PheOMe. Trypsin potentiated the inhibitory effect of PheOMe on NK activity and monocyte depletion. Trypsin partially inhibited IL-2 activation of LAK activity enhanced by PheOMe. Chymotrypsin had little effect on NK activity but prevented the inhibitory effect of PheOMe on NK activity. It had little effect on monocyte depletion induced by PheOMe. PheOMe was hydrolysed by monocytes and chymotrypsin to Phe and methanol as determined by HPLC. TPCK inhibited hydrolysis of PheOMe by monocytes. Our data suggest that the effects of PheOMe on monocytes, NK cells and LAK activation involve protease activities of monocytes. 相似文献
172.
G. Akoev G. N. Andrianov T. Szabo B. Bromm 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1991,168(6):639-646
Summary Dissected ampullae of Lorenzini of the skate (Raja clavata) were studied with the aim of determining the synaptic transmitter between electroreceptor cell and afferent fibre. Resting activity and stimulus-evoked activity in response to electrical pulses were recorded in single afferent units at constant perfusion with normal and test solutions containing different putative neurotransmitters. Presynaptic transmitter release was blocked by Mg2+ (up to 50 mM) to investigate the effects of the test substances upon the postsynaptic membrane. l-Glutamate (l-GLU) and l-aspartate (l-ASP), both at concentrations between 10-7 and 10-3
M, enlarged strongly resting and stimulus-evoked discharge frequency in the afferent fibre. If transmission was blocked by high Mg2+, resting discharge frequency could be restored by l-GLU or l-ASP. The glutamate agonists quisqualate (10-8–105
M) and N-methyl-D-aspartate (10-5–10-3
M) enlarged spontaneous activity in the afferent fiber. The same was found for kainic acid (10-9–10-5
M). Taurine at concentrations between 10-5 and 10-3
M caused a concentration-dependent decrease in afferent activity. The same was found for gammaaminobutyric acid (GABA; 10-5–10-4
M), and for the catecholamines adrenaline and noradrenaline, both in concentrations between 10-5 and 10-3
M. Serotonine (10-5–10-3
M) and dopamine (10-5-10-3
M) had no effect on resting or evoked activity in the Lorenzinian ampulla afferents. Acetylcholine (ACh; 10-4
M) enlarged discharge frequency in those units with initial rates lower than 22–25 Hz, but diminished discharge frequency in fibres with initial activity higher than 25 Hz. When synaptic transmission was blocked by high Mg2+ solution, perfusion with additional ACh did not restore resting activity in the afferent fibre. The results suggest that the most probable transmitter in the afferent synapse of the ampullae of Lorenzini is l-GLU or l-ASP, or a substance of similar nature.Abbreviations
ACh
acetylcholine
-
GABA
gamma aminobutyric acid
-
KA
kainic acid
-
l-ASP
l-aspartate
-
l-GLU
l-glutamate
-
NMDA
N-methyl-D-aspartate
-
Q
quisqualate
-
n.s.
normal solution 相似文献
173.
In the internal granular layer of the cerebellar cortex the polysynaptic complexes called glomeruli consist mainly of homogeneous populations of glutamatergic and GABAergic synapses, both located on granule cell dendrites. A subcellular fraction enriched in glomeruli was prepared from rat cerebellum, and the distribution of the different types of NMDA and non-NMDA glutamate binding sites was studied in the membranes derived from this fraction (fraction G) as compared to that in the membranes prepared from a total cerebellar homogenate (fraction T). Cl–/Ca2+ independent [3H]glutamate binding sites were not abundant and could be reliably measured only in fraction G. Cl– dependent/Ca2+ activated [3H]glutamate binding sites were more abundant and exhibited a single K
d
in both fractions G and T. Quisqualate, NMDA, kainate, L-AP4 andtrans-ACPD inhibited [3H]glutamate binding to different extents in the two membrane fractions. Quisqualate sensitive sites were predominant in all cases but more abundant in fraction T than in fraction G. An opposite distribution was observed for the NMDA sensitive binding sites while kainate sensitive binding sites were scarce everywhere.Trans-ACPD, a ligand presumed selective for metabotropic glutamate binding sites, displaced [3H]glutamate from fraction T but nor from fraction G, suggesting the absence of these sites from glomeruli. Similarly, no L-AP4 sensitive sites were present in fraction G while they were abundant in fraction T. Binding sites associated with ionotropic receptors of the quisqualate type were determined by measuring [3H]AMPA binding. The density of the high affinity [3H]AMPA binding sites in fraction T was twice as high as in fraction G, indicating that these sites are abundant in structures other than glomeruli. High-affinity [3H]kainate binding sites are more abundant in fraction G than in fraction T; the same, but with smaller differences, occurs for the distribution of the low affinity [3H]kainate binding sites. The density of the latter sites is close to that of the high affinity [3H]AMPA binding sites confirming the presence of quisqualate/kainate receptors on granule cells, as previously hypothesized (for review, see Gallo et al., 1990). Taken together, these results indicate a segregation of the glutamate binding sites types at specialized synapses or neuronal cell types in the cerebellar network.Abbreviations AMPA
(RS)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid
- DL-AP4
dl-2-amino-4-phosphonobutyric acid
- D-AP5
d-2-amino-5-phosphonovaleric acid
- EAA
excitatory amino acid
- EGTA
ethylene glycol-bis(-aminoethyle ether) N,N,N,N-tetracetic acid
- NMDA
N-methyl-D-aspartate
- Quisqualate
-[3,5-dioxo-1,2,4-oxadiazolidin-2-yl]-L-alanine
-
trans-ACPD
trans-1-amino-cyclopentyl-1,3-dicarboxylic acid 相似文献
174.
Summary A re-examination of earlier NPK fertilization experiments in Douglas fir stands on sandy soils shows the effects of high nitrogen
input by air pollution during the last 10–15 years on plant nutrition at these sites. In 1960, experimental plots showed a
positive growth reaction to nitrogen, phosphorus, and potassium fertilization. All suffered from severe phosphorus deficiency
in 1984, low phosphorus in the needles was invariably accompanied by a high nitrogen content, with all N/P ratios between
20 and 30. The same conclusion emerges from an independent investigation of nutrient status of a selection of Douglas fir
stands. Hence, if stand productivity and a balanced nutrient status of the trees is to be maintained, the increase in atmospheric
input of nitrogen calls for supplementary fertilization. Given the current N/P ratios in the needles, a positive growth response
to phosphorus fertilization is to be expected. 相似文献
175.
Summary Nitrate fertiliser labelled with15N was applied to a field grown crop of winter wheat. Uptake and assimilation of fertiliser nitrate was studied by monitoring the appearance of labelled nitrate and labelled amino acids in the xylem sap. Shortly after applying15N-nitrate to the soil about 30 per cent of recently absorbed15N was in the reduced form, indicating that roots of cereal crops can make a substantial contribution in reducing nitrate. Seasonal changes in crop growth andin vivo NRA are also described. 相似文献
176.
Summary Ammonium nitrate fertilizer, labelled with15N, was applied in spring to winter wheat growing in undisturbed monoliths of clay and sandy loam soil in lysimeters; the rates
of application were respectively 95 and 102 kg N ha−1 in the spring of 1976 and 1975. Crops of winter wheat, oilseed rape, peas and barley grown in the following 5 or 6 years
were treated with unlabelled nitrogen fertilizer at rates recommended for maximum yields. During each year of the experiments
the lysimeters were divided into treatments which were either freelydrained or subjected to periods of waterlogging. Another
labelled nitrogen application was made in 1980 to a separate group of lysimeters with a clay soil and a winter wheat crop
to study further the uptake of nitrogen fertilizer in relation to waterlogging.
In the first growing season, shoots of the winter wheat at harvest contained 46 and 58% of the fertilizer nitrogen applied
to the clay and sandy loam soils respectively. In the following year the crops contained a further 1–2% of the labelled fertilizer,
and after 5 and 6 years the total recoveries of labelled fertilizer in the crops were 49 and 62% on the clay and sandy loam
soils respectively.
In the first winter after the labelled fertilizer was applied, less than 1% of the fertilizer was lost in the drainage water,
and only about 2% of the total nitrogen (mainly nitrate) in the drainage water from both soils was derived from the fertilizer.
Maximum annual loss occurred the following year but the proportion of tracer nitrogen in drainage was nevertheless smaller.
Leaching losses over the 5 and 6 years from the clay and sandy loam soil were respectively 1.3 and 3.9% of the original application.
On both soils the percentage of labelled nitrogen to the total crop nitrogen content was greater after a period of winter
waterlogging than for freely-drained treatments. This was most marked on the clay soil; evidence points to winter waterlogging
promoting denitrification and the consequent loss of soil nitrogen making the crop more dependent on spring fertilizer applications. 相似文献
177.
E. Wada R. Imaizumi Y. Kabaya T. Yasuda T. Kanamori G. Saito A. Nishimune 《Plant and Soil》1986,93(2):269-286
Summary Plants from agricultural and natural upland ecosystem were investigated for15N content to evaluate the role of symbiotic N2-fixation in the nitrogen nutrition of soybean. Increased yields and lower δ15N values of nodulating soybeansvs, non-nodulating isolines gave semi-quantitative estimates of N2 fixation. A fairly large discrepancy was found between estimations by δ15N and by N yield at 0 kg N/ha of fertilizer. More precise estimates were made by following changes in plant δ15N when fertilizer δ15N was varied near15N natural abundance level. Clearcut linear relationships between δ15N values of whole plants and of fertilizer were obtained at 30 kg N/ha of fertilizer for three kinds of soils. In experimental
field plots, nodulating soybeans obtained 13±1% of their nitrogen from fertilizer, 66±8% from N2 fixation and 21±10% from soil nitrogen in Andosol brown soil; 30%, 16% and 54% in Andosol black soil; 7%, 77% and 16% in
Alluvial soil, respectively. These values for N2 fixation coincided with each corresponding estimation by N yield method.
Other results include: 1)15N content in upland soils and plants was variable, and may reflect differences in the mode of mineralization of soil organics,
and 2) nitrogen isotopic discrimination during fertilizer uptake (δ15N of plant minus fertilizer) ranged from −2.2 to +4.9‰ at 0–30 kg N/ha of fertilizer, depending on soil type and plant species.
The proposed method can accurately and relatively simply establish the importance of symbiotic nitrogen fixation for soybeans
growing in agricultural settings. 相似文献
178.
Summary Remobilization of15N from vegetative tissue of mungbean (Vigna radiata (L.) Wilczek) into pods was measured during the reproductive phase of growth. Plant tissue was labelled with15N during vegetative development. Experiments were conducted in the field at two sites. At one site the soil provided cowpeas
with most of their N but at the other site N fixation provided most of the N. Remobilized N from vegetative tissue to pods
occurred soon after they began to develop. The quantity of the labelled N ultimately remobilized to the pods amounted to 50%
for one cultivar (Tx33) at the high soil N site and 70% at the low N site. For the other cultivar (Tx13) the values were 25%
and 30%, respectively. The two cultivars performed very differently with respect to partitioning of N into pods and the rate
of N fixation. Even though more N was accumulated in the shoots of the high N fixing cultivar (Tx13) less total N was contained
in the pods. 相似文献
179.
Summary Non-symbiotic heterotrophic N2 fixation in coniferous bark litter was investigated with the acetylene reduction assay under aerobic and anaerobic conditions. The litter studied was composed essentially of bark, of pH 5 and a C/N ratio of 101; the ratio of available C to available N, which governs N2 fixation, was considerably higher. The rate of N2 fixation was estimated as 2.5–4.4 g N. g–1 dry wt. day–1. Nitrogenase activity was still evident after seven months of incubation under aerobic conditions. The N2-ase activity was O2 dependent: under anaerobic conditions no N2-ase activity was found unless a fermentable C source was added. The importance of N2 fixation in N-poor litter for the maintenance of soil fertility is emphasized. 相似文献
180.
An Escherichia coli mutant unable to support site-specific recombination of bacteriophage lambda 总被引:23,自引:0,他引:23
We report the isolation of mutations in, and the characterization of, an Escherichia coli gene, hip, that is required for site-specific recombination of phage lambda. hip mutants are recessive and are located near minute 20 on the linkage map. The gene product is not vital to bacterial growth, since deletion mutants are viable. The absence of hip product reduces lambda integration to barely detectable levels and also reduces prophage excision, but less drastically. Certain mutations in the lambda int gene partially restore integration and excision in hip- hosts. Homologous recombination promoted by recA does not require hip function. In addition to their defect in site-specific recombination, hip mutants are unable to support lytic growth of phage Mu or of certain lambda mutants. Their pleiotropic phenotype closely resembles that of himA mutants, but complementation, mapping and DNA sequencing show that hip and himA are different genes. 相似文献