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211.
Vicent Casadó Carmen Lluis Enrique Canela Rafael Franco Josefa Mallol 《Neurochemical research》1992,17(2):129-139
Pig brain cerebral cortex was subfractionated by isopycnic centrifugation in sucrose gradients. In each subfraction the content of the agonist [3H]R-PIA binding, the activity of adenosine metabolizing enzymes (5-nucleotidase and adenosine deaminase) and the activity of membrane marker enzymes were determined. The fractions were also examined by electron microscope. In general, the results suggest a widespread distribution of A1 adenosine receptors in membranes from different origins. Marker enzyme profile characterization indicated an enrichment of A1 adenosine receptor in pre-synaptic membranes isolated from the crude synaptosomal fraction (P2B subfraction) as well as in membranes of glial origin such as myelin. The receptor is also present in the endoplasmic reticulum and in membranes isolated from the microsomal fraction that seem to have a post-synaptic origin (P3B). In subfractions having a high content of adenosine receptor the equilibrium binding paramters were obtained as well as the proportion of high- to low-affinity sites. From the values of the equilibrium constants it was not possible to find differences between the receptor in the different subfractions. Analysis of the affinity state distribution showed a diminished percentage of high-affinity sites in fraction P3A, which can be accounted by the existence of myelin membranes; in contrast the percentage of high-affinity states was higher in P2 and P3B, indicating that in these fractions the receptor is present in synaptosomal membranes. The close correlation shown between the enzyme 5-nucleotidase specific activity and the specific ligand binding distributions led us to postulate an important role for the enzyme in the regulation of adenosine action in pig brain cortex. 相似文献
212.
Mineralization budgets in sediment microcosms: Effect of the infauna and anoxic conditions 总被引:1,自引:0,他引:1
Abstract A number of sediment incubations were set up to reproduce some of the conditions used by Kristensen and Blackburn [1] and to make a comparison with their results. There were three types of microcosm: aerobic (OX), anaerobic (AN) and aerobic with Nephtys (NOX). In addition to other measurements, dissolved organic nitrogen (DON) pools and fluxes, were measured. The sediment in this experiment contained more particulate organic matter (POM). Nephtys (NOX) had the same effect as Nereis in increasing the rate of mineralization of POC and PON, compared with the OX-cores (2.1 and 2.6 times, respectively). Again, the AN-cores had a higher mineralization rate (loss of POM) than that of the OX-cores, but in addition, mineralization in NOX-cores was not significantly different from AN-cores. It was thus confirmed that anoxic mineralization could be as high, or higher, than the oxic process. Both the temporal patterns of O2 -and and CO2 -fluxes and their magnitudes were very similar to those reported earlier. This contrasts with the higher loss of POM in the present experiment. However, the loss of C in DOC (associated with the measured DON) can account for the extra POM loss. The pore-water profiles of σCO2 and NH4 + were similar to those in the earlier report, and the fluxes of σCO2 , O2 , NH4 + and NO3 − followed the same temporal pattern. 相似文献
213.
The genetic structure of six populations of Iran (Turks, Kurds, Lurs, Zabolis, Baluchis and Zoroastrians) was examined using
data on blood groups, serum proteins and cell enzymes. Our results show conclusively that there are genetic differences among
the six populations and the analysis of superimposed R and S matrices defined by Harpending & Jenkins (1973) show that the
dispersion of some of the alleles correspond to the dispersion of the populations. The FST estimates are not large enough to favour selection on any of the loci studied. The FIT and FIS estimates are positive and moderately high suggesting that the genetic differentiation to some extent is influenced by inbreeding. 相似文献
214.
Stefan Jansson Eran Pichersky Roberto Bassi Beverley R. Green Masahiko Ikeuchi Anastasios Melis David J. Simpson Michael Spangfort L. Andrew Staehelin J. Philip Thornber 《Plant Molecular Biology Reporter》1992,10(3):242-253
We propose a nomenclature for the genes encoding the chlorophylla/b-binding proteins of the light-harvesting complexes of photosystem I and II. The genes encoding LHC I and LHC II polypeptides
are namedLhca1 throughLhca4 andLhcb1 throughLhcb6, respectively. The proposal follows the general format recommended by the Commision on Plant Gene Nomenclature. We also present
a table for the conversion of old gene names to the new nomenclature. 相似文献
215.
Limitations of the scrape-loading/dye transfer technique to quantify inhibition of gap junctional intercellular communication 总被引:5,自引:0,他引:5
Gap junctional intercellular communication (GJIC) is recognized as playing an important role in normal cell proliferation and development. Chemically induced alteration of GJIC has been proposed to be associated with abnormal cellular growth and/or tumor promotion. Several in vitro assays are currently used to determine the effects of chemicals on GJIC between cultured mammalian cells. One of these assays, the scrape-loading dye transfer (SLIDT) technique, is based on monitoring the transfer of the fluorescent dye Lucifer yellow from one cell into adjacent cells via functional gap junctions. The objective of our study was to evaluate and compare various approaches for quantifying results obtained with the SL/DT technique. Confluent cultures of either WB rat liver epithelial cells or LC-540 rat leydig cells were exposed to the animal tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), solvent (0.1% ethanol), or culture medium for one hour at 37° C prior to analysis of GJIC. Inhibition of dye transfer was clearly evident following TPA exposure. Quantification of this dye transfer was assessed via four approaches: manually counting the number of labeled cells; measuring the distance of dye travel from the scrape line; quantifying the amount of cellular dye uptake; and determining the distribution of dye away from the scrape line. Our results suggest that while the SL/DT technique can be effectively used as a tool to determine the qualitative presence or absence of GJIC, its use in quantifying changes in GJIC following chemical exposure is limited. Since concentration-dependent responses are critical in chemical testing, application of the SLIDT method should be restricted to a screening assay for qualitatively assessing the presence or absence of GJIC. Another assay (e.g., electrical coupling, microinjection, metabolic cooperation, radioactive metabolite transfer, or fluorescence redistribution after photobleaching) should be considered to quantify changes in GJIC and construct chemical concentration-response curves.Abbreviations FBS,
fetal bovine serum
- GJIC,
gap junctional intercellular communication
- HBSS,
Hank's balanced saline solution
- SL/DT,
scrape-loading/dye transfer
- TPA,
12-O-tetradecanoylphorbol-13-acetate. 相似文献
216.
The packing of peptide helices in crystals of the leucine-rich decapeptide Boc-Aib-Leu-Aib-Aib-Leu-Leu-Leu-Aib-Leu-Aib-OMe provides an example of ladder-like leucylleucyl interactions between neighboring molecules. The peptide molecule forms a helix with five 5----1 hydrogen bonds and two 4----1 hydrogen bonds near the C terminus. Three head-to-tail NH ... O = C hydrogen bonds between helices form continuous columns of helices in the crystal. The helicial columns associate in an antiparallel fashion, except for the association of Leu ... Leu side chains, which occurs along the diagonal of the cell where the peptide helices are parallel. The peptide, with formula C56H102N10O13, crystallizes in space group P2(1)2(1)2(1) with Z = 4 and cell parameters a = 16.774(3) A, b = 20.032(3) A and c = 20.117(3) A; overall agreement factor R = 10.7% for 2014 data with magnitude of F(obs) greater than 3 sigma (F); resolution 1.0 A. 相似文献
217.
Francisco J. Acosta Jose M. Serrano Jesus M. Barandica Francisco Lopez 《Journal of Insect Behavior》1992,5(3):337-346
The focal-animal technique assumes the continuous recording of the behavior of an individual during a certain time interval. The length of this interval (sampling unit) can be problematic when estimating the duration of behavioral acts. Two acts from the behavioral repertoire of the ant Leptothorax fuenteiwere focused on in this work at different ranges of temporal scales. Analyzing these acts we show the possibility of existence of a sampling artifact, in such a way that the estimates of the durations of the acts would be forced to depend upon the length of the sampling unit that is being used. 相似文献
218.
J. H. Medina C. Peña M. Piva C. Wolfman M. L. de Stein C. Wasowski C. Da Cunha I. Izquierdo A. C. Paladini 《Molecular neurobiology》1992,6(4):377-386
Great progress has been made in the last 5 yr in demonstrating the presence of benzodiazepines (BDZs) in mammalian tissues,
in beginning studies on the origin of these natural compounds, and in elucidating their possible biological roles. Many unanswered
questions remain regarding the sources and biosynthetic pathways responsible for the presence of BDZs in brain and their different
physiological and/or biochemical actions. This essay will focus on recent findings supporting that: (1) BDZs are of natural
origin; (2) mammalian brain contains BDZs in concentrations ranging between 5.10−10–10−8
M; (3) dietary source of BDZs might be a plausible explanation for their occurrence in animal tissues, including man; (4) the
formation of BDZ-like molecules in brain is a possibility, experimentally supported; (5) BDZ-like molecules including diazepam
andN-desmethyldiazepam are elevated in hepatic encephalopathy; and (6) natural BDZs in the brain are involved in the modulation
of memory processes. Future studies using the full range of biochemical, physiological, behavioral, and molecular biological
techniques available to the neuroscientist will hopefully continue to yield exciting and new information concerning the biological
roles that BDZs might play in the normal and pathological functioning of the brain. 相似文献
219.
Regarding biological products, increasing awareness of potential side effects have placed great importance not only at protein
purity regarding other proteins but on the removal of biologicals such as DNA and especially virus the importance of which
may not be known. Monoclonal antibodies (Mab) have come to be an important class of molecules obtained from hybridoma cells,
i.e., nonrecombinant cells in culture. It has been noted during the last years, that with rare exceptions hybridoma cell lines
contain retrovirus like particles. The infectious nature of the EM-visible particles has been tested for, however, in most
cases not been substantiated. In order to bring these valuable biological reagents, Mab's, to good use in man for imaging
or therapy, the remaining concern about a potential retroviral infection has to be reduced to an acceptable minimum. We describe
experimental approaches for the validation of chromatographic and ultrafiltration steps used in the production of monoclonal
antibodies to remove and inactivate murine retrovirus.
Present day biotechnological manufacturing processes have been devised incorporating a number of strategic preventive measures
that have found wide spread acceptance. They permit to answer the question: how can a potentially harmful infection by an
unknown virus be excluded.
Knowledge of the efficacy of purification steps to clear infectious model virus is fundamental to devise biotechnological
manufacturing processes yielding a purified antibody for use in man. 相似文献
220.
Summary Most of the inducible mutagenesis observed in Escherichia coli after treatment with many DNA damaging agents is dependent upon the products of the umuD,C operon. RecA-mediated proteolytic processing of UmuD yields a carboxyl-terminal fragment (UmuD) that is active for mutagenesis. Processing of UmuD is therefore a critical step in the fixation of mutations. In this paper we have analyzed the requirements for UmuD processing in vivo. Standard immuno-detection assays, coupled with a sensitive chemiluminescence detection assay, have been utilized to probe levels of chromosomally encoded Umu proteins from whole-cell E. coli extracts. We found that the derepression of additional SOS gene products, other than RecA, was not required for UmuD processing. Moreover, efficient cleavage of UmuD was observed only in the presence of elevated levels of activated RecA, suggesting that efficient processing would occur only under conditions of severe DNA damage. Detection of chromosomally encoded Umu proteins has allowed us, for the first time, to measure directly the cellular steady-state levels of these proteins under various SOS inducing conditions. UmuD was present at 180 copies per uninduced cell and was measured at 2400 copies per cell in strains that lacked a functional repressor. Induced levels of UmuC were approximately 12-fold lower than UmuD with 200 molecules per cell. These levels of cellular UmuC protein suggest that it functions through specific protein-DNA or protein-protein interactions, possibly as a lesion recognition protein or by interacting with DNA polymerase III. 相似文献