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T E Webb

Publications and source records attributed to T E Webb.

At least 109 records · Page 6Linked to original sources
PubMed1979

Heterogeneity and properties of hepatic dexamethasone-binding proteins.

Evidence from experiments in vivo and in vitro is presented for the presence of three species of dexamethasone-binding proteins in rat liver, which are identified by chromatography on Sepharose 6B or by isoelectric focusing. Although two of these species (DI and DII) possess properties characteristic of a true receptor, the third binding protein (i.e. DIII), which migrates most slowly on Sepharose 6B, but has stability properties similar to protein DII, exhibits a 3-fold lower affinity for dexamethasone and the activated complex neither binds to DNA-cellulose nor translocates to the nucleus. Only the predominant liver receptor (DI), which is eluted first from Sepharose 6B, is present in Novikoff-hepatoma cytosol, suggesting that the major and minor species are not interconverted through simple dissociation during their isolation. The binding activities of all three species in the liver cytosol increase approx. 2-fold in vivo after adrenalectomy and show a transient 2-fold fall in vivo after the administration of cortisol. These changes in vivo in protein DIII shows a marked lag compared with those in proteins DI and DII, which change in parallel. It is therefore proposed that rat liver cytosol contains two dexamethasone receptors and a dexamethasone-binding protein that may be derived from these receptors.

Adrenalectomy↗
PubMed1979

Uridine kinase activity in human tumors.

The activity of uridine kinase, a key enzyme in the salvage pathway for pyrimidine bases and nucleosides and for the activation of the corresponding chemotherapeutic analogs, varied 10-fold in a series of human colon adenosarcomas; similar variations were observed with other tumor types. In contrast to the leukemias where only the adult isozyme appears to be present, both the adult and embryonic forms were present in the solid tumors examined. The qualitative and quantitative differences may account, in part, for differences in the innate (initial) sensitivity of the tumors to pyrimidine base and nucleoside analogs.

Animals↗
PubMed1978

Effect of adenosine 3':5'-monophosphate and guanosine 3':5'-monophosphate on RNA release from isolated nuclei.

The addition of physiological concentrations of either cAMP or cGMP stimulated the release of RNA from isolated prelabeled rat liver nuclei to a fortified cytosol in a cell-free system. The released RNA was shown to be primarily mRNA by its binding to oligo(dT)-cellulose and its sedimentation profile. Treatment of rats with cAMP or cGMP 30 min prior to the preparation of cyclic nucleotides on the cell-free system. Cyclic nucleotides stimulation of RNA release occurred in systems prepared from resting rat liver, Novikoff hepatoma, and Morris hepatoma 5123D, but not the 18-h regenerating liver. The response of the cell-free system to added cyclic nucleotides reflected the in vivo concentration of these substances in the tissues from which the system was prepared. Those with high in vivo levels were not stimulated while those with lower levels did respond to added cyclic nucleotides. Neither cAMP nor cGMP had an appreciable effect on rRNA release.

Animals↗
PubMed1977

Uridine kinase activities and pyrimidine nucleoside phosphorylation in fluoropyrimidine-sensitive and -resistant cell lines of the Novikoff hepatoma.

Uridine kinase, the rate-limiting enzyme in the activation (phosphorylation) of uridine and the corresponding chemotherapeutic analogues, is present as two isoenzymes localized exclusively in the cytosol of rapidly growing neoplasms, including the S-37 sarcoma, EL-4 leukaemia, HeLa cells (a human carcinoma) and the Novikoff hepatoma. The activities of the isolated isoenzymes are markedly decreased when the concentrations of ATP, phosphate or Mg2+ that are optimum in vitro are replaced by concentrations of ATP, phosphate or Mg2+ that are optimum in vitro are replaced by concentrations approximating to those found in vivo. Further, comparisons of the Km values of isolated uridine kinases with those for cellular uptake of pyrimidine nucleosides and their rate of intracellular phosphorylation suggest that nucleoside-transport systems play a rate-limiting role in nucleoside analogue activation and consequently that it is impossible to estimate the Km of uridine kinase in the intact cell. During the development of tumour-cell resistance to 5-fluorouracil or 5-fluorouridine in vivo there was an early differential increase in the activity of a low-affinity (high-Km) uridine kinase isoenzyme, as measured in cell extracts, and a 7-fold increase in the Km values for the uptake of both uridine and 5-fluorouridine into the intact resistant cells.

Animals↗
PubMed1977

Proflavine sensitivity of RNA processing in isolated nuclei.

The intercalating agent proflavine inhibits the processing and subsequent release of preformed messenger RNA and ribosomal RNA from isolated liver nuclei to surrogate cytoplasm. The direct effect of proflavine on these processes, as monitored in a reconstituted cell-free system, supports the theory that base-paired segments (i.e. hairpin loops) in the precursor RNA's are involved as recognition sites in nuclear RNA processing.

Acridines↗
PubMed1977

Elevated concentration of a dexamethasone-receptor translocation inhibitor in Novikoff hepatoma cells.

Despite the apparent similarity of the dexamethasone-receptor concentration in the rat liver and Novikoff hepatoma cytoplasms, the in vitro translocation of the dexamethasone-receptor complex into liver or hepatoma nuclei was three times greater from liver as compared to hepatoma cytosol; both cytosols showed an apparent saturation of nuclear receptor sites. Although the competitive interaction of the receptor complex with nuclei suggested that they were qualitatively similar, the translocation of additional dexamethasone-receptor from liver cytosol into nuclei presaturated with the receptor complex from hepatoma cytosol suggested that they were different. However, the latter observation, as well as the initial differential nuclear uptake of the dexamethasone-receptor complex from liver and hepatoma cytosols, can be accounted for by a higher concentration of a translocation inhibitor in the hepatoma cytosol. Thus hepatoma cytosol, at a protein concentration of 6 mg/ml and free of activated receptor complex, was four times more inhibitory to nuclear uptake of the activated complex than was a comparable preparation of liver cytosol. A study of the kinetics of nuclear uptake of the partially purified dexamethasone-receptor complex from hepatoma confirmed that, in the absence of cytoplasmic translocation inhibitor, nuclear acceptor sites were not limiting in vitro. Evidence is presented which suggests, but does not prove, that the cytoplasmic translocation inhibitor functions as such in the intact cell.

Animals↗
PubMed1977

Uridine kinase activities in normal and neoplastic lymphoid cells.

Both adult (I) and embryonic (II) forms of uridine kinase have been identified in the transplantable EL-4 leukemia of C57BL/6 mice and in the P815Y mastocytoma of DBA/2 mice. Only Species I is found in primary tumor cells of lymphoid orgin (virus-induced feline lymphosarcoma, human acute and chronic lymphocytic leukemia) and in normal calf thymocytes and porcine peripheral blood lymphocytes; Species I was induced 4-fold upon stimulation of the normal blood lymphocytes with phytohemagglutinin. The level of uridine kinase activity in the feline lymphosarcoma of thymus-dependent lymphocyte orgin and childhood lymphocytic leukemia of possible thymus-dependent lymphocyte or null-cell origin was similar to the induced level in phytohemagglutinin-stimulated normal lymphocytes, i.e., thymus-dependent lymphocytes. In contrast lymphocytes of a patient with chronic lymphocytic leukemia of thymus-independent lymphocyte origin had a level of uridine kinase activity comparable to that of the unstimulated normal lymphocytes or thymocytes. The uridine kinase activity in the EL-4 tumor cells was repressed by acute treatment of the mice with 5-azacytidine.

Animals↗
PubMed1976

Nature of the facilitated messenger ribonucleic acid transport from isolated nuclei.

Cytoplasmic macromolecules were previously identified which regulate both qualitatively and quantitatively the release of messenger-like RNA from isolated nuclei. These macromolecules are now shown to be denatured at 45-50 degrees C and their synthesis is sensitive to pactamycin or cycloheximide. The putative regulatory proteins are essentially quantitatively precipitated with high specificity from the cytosol by streptomycin at a concentration 10-fold higher than that used to precipitate RNA. The nuclear concentration-dependence of RNA transport from successive samples of nuclei strongly suggests that the regulatory factors are recycled. Quantitative changes in the sequences transported at various dilutions of the cytosol suggest that not all the different classes of the putative regulatory macromolecules are present in an effective concentration at any one dilution.

Animals↗
PubMed1976

The repression and derepression of hepatic tyrosine aminotransferase by carcinogens.

Like hydrocortisone, a single carcinogenic dose of dimethylnitrosamine (50 mg/kg) initiates the induction cycle for hepatic tyrosine aminotransferase in adrenalectomized rats. However, following this initial induction in the presence of dimethylnitrosamine, the enzyme becomes refractory to reinduction by known inducers. The administration of thioacetamide to either adrenalectomized or intact rats leads to an immediate and progressive loss of inducibility by hydrocortisone, dibutyrylcyclic AMP or dimethylnitrosamine. Although the thioacetamide-induced repression was not reversed even up to 10 weekds after the cessation of treatment, it was reversed after the induction of liver regeneration. Both the carcinogen-mediated induction and repression of tyrosine aminotransferase appears to occur by mechanisms which do not involve the corticosteroid-binding proteins which normally mediate the induction by glucocorticoids.

Acetamides↗
PubMed1976

In vivo synchronization of murine sarcoma 37 ascites tumor cells by use of hydroxyurea.

Synchrony of DNA synthesis in mouse sarcoma 37 ascites cells was achieved in vivo by injection of 0.75 mg hydroxyurea per gram weight of mouse. DNA synthesis was gauged by comparing 3H-labeled thymidine incorporation, under defined conditions, to total DNA measured colorimetrically. DNA synthesis in the sarcoma 37 cells fell to negligible levels 4 hours after injection and remained at or below levels observed in randomly dividing cells up to 18 hours after injection. A sharp increase in DNA synthesis occurred at 20 hours, followed by a decline again at 22 hours after hydroxyurea injection. DNA synthesis in the intestine remained below control values at 20 hours.

Animals↗